Staphylococcus aureus

Infect Immun, 1987 Dec, 55(12), 3103 - 10
Virulence of protein A-deficient and alpha-toxin-deficient mutants of Staphylococcus aureus isolated by allele replacement; Patel AH et al.; The gene coding for protein A (spa) of Staphylococcus aureus 8325-4 has been inactivated by substituting part of the spa coding sequence for a DNA fragment specifying resistance to ethidium bromide . The in vitro-constructed spa::EtBrr substitution mutation was introduced into the S . aureus chromosome by recombinational allele replacement . Southern blot hybridization showed that the in vitro-constructed mutation was present in the chromosomal spa locus . We have previously reported the inactivation of the alpha-toxin gene (hly) by allele replacement with an in vitro-constructed hly::Emr (erythromycin resistance) mutation (M . O'Reilly, J.C.S . de Azavedo, S . Kennedy, and T.J . Foster, Microb . Pathogen . 1:125-138, 1986) . A double Spa- Hly- mutant was constructed by transduction . The virulence of Spa- and Hly- mutants was tested by experimental infection of mice . When subcutaneous injections were given, Hly- mutants formed a flat, darkened lesion, whereas Hly+ strains caused a raised, cream lesion . Alpha-toxin was shown to be a major factor in forming subcutaneous lesions and in causing the death of mice injected intraperitoneally . Spa- mutants were slightly less virulent than their Spa+ counterparts, which suggests that protein A is also a virulence factor of S . aureus.

Infect Immun, 1987 Dec, 55(12), 3030 - 4
Influence of Staphylococcus aureus antibody on experimental endocarditis in rabbits; Greenberg DP et al.; To evaluate the potential protective benefit of antibody to whole cells of Staphylococcal aureus for the prevention of endocarditis, the rabbit endocarditis model was used . Methicillin-sensitive (17A) and methicillin-resistant (173) S . aureus strains were evaluated in rabbits with or without indwelling intracardiac catheters . All immunized rabbits developed significant homologous agglutinating antibody titers (the mean reciprocal titers were 15,300 to strain 17A and 1,150 to strain 173) . After challenge, virtually no significant differences were observed between immunized and unimmunized animals with respect to (i) incidence of endocarditis, (ii) concentration of bacteria in infected vegetations, (iii) incidence of metastatic renal abscesses, or (iv) concentrations of bacteria in infected kidneys . The clearance of homologous S . aureus strains from blood cultures was similar for immunized and unimmunized animals at 10 to 90 min after intravenous challenge . In vivo adherence of homologous S . aureus strains to aortic valves and vegetations was similar in immunized and unimmunized animals when evaluated at 30 and 90 min postchallenge . Even without catheterization, the incidence of bacteremia and renal abscesses was the same in immunized and unimmunized rabbits . Whole-cell-induced S . aureus antibody did not prevent or modify any stage in the development of endocarditis in rabbits.

Clin Orthop, 1987 Dec, (225), 234 - 7
Disc space infection and vertebral osteomyelitis as a complication of percutaneous lateral discectomy; Blankstein A et al.; Percutaneous lateral discectomy (PLD) in a 32-year-old man was followed by postoperative disc space infection and adjacent vertebral osteomyelitis caused by Staphylococcus aureus . The simplicity and decreased morbidity associated with PLD may be offset by severe infections . The small incision made in the annulus during PLD may not allow adequate drainage in the case of infection and may subsequently direct the infective process to the adjacent vertebral endplates . Meticulous aseptic technique, and possibly the use of prophylactic antibiotic therapy, is important in PLD.

Zentralbl Bakteriol Mikrobiol Hyg {A}, 1987 Dec, 267(2), 167 - 72
Location of the coagulase gene in Staphylococcus aureus; Sawicka-Grzelak A et al.; The localization of the coagulase gene in Staphylococcus aureus JM 750 strain was investigated . S . aureus JM 750 strain in contrast to other S . aureus strains shows instability of coagulase production . This strain contains the 23.5 kb penicillinase plasmid, determining beta-lactamase synthesis and resistance to heavy metal salts, and a larger plasmid greater than 100 kb in size, which was detectable only in coagulase positive variants . The results suggest the unparalleled, extrachromosomal localization of the coagulase gene in S . aureus JM 750 strain.

Clin Exp Immunol, 1987 Dec, 70(3), 649 - 57
Inhibition of T cell-dependent human B cell proliferation and B cell differentiation by polyspecific monomeric IgG; Stohl W et al.; A commercially available polyspecific, monomeric IgG preparation suitable for intravenous administration (IgSRK; Sandoglobulin) can inhibit pokeweed mitogen (PWM)-induced proliferation of peripheral blood mononuclear cells (PBMC) by a small, but statistically significant, amount compared to control cultures . Such inhibition could not be demonstrated when PBMC were stimulated with the T cell mitogen phytohaemagglutinin . Surface phenotype analysis of the PWM-stimulated cells indicated that in IgSRK-containing cultures, the proportion of B cells was decreased and the proportion of T cells was increased compared to control cultures . This alteration in T:B ratio was not due to antigenic modulation of B or T cell markers from their surfaces . In addition, IgSRK inhibited the proliferation of T cell-depleted PBMC cultures stimulated by B cell proliferation factors (BCPF) but not by fixed protein A-bearing Staphylococcus aureus strain Cowan I . The capacity to inhibit B cell proliferation was independent of and distinct from its capacity to inhibit B cell differentiation, since IgSRK inhibited the differentiation of a B cell differentiation factor (BCDF)-sensitive line by BCDF (which contains no BCPF activity) . IgSRK inhibited PWM-induced generation of cytoplasmic Ig+ cells but had no effect on Ig secretion from mature Ig-secreting cells . Taken together, these findings suggest that IgSRK (which contains the IgG fraction from pooled plasma from 2,000 healthy donors) can inhibit T cell-dependent or T cell factor-dependent B cell proliferation and B cell differentiation.

Mol Immunol, 1987 Dec, 24(12), 1273 - 80
Two novel phospholipid-linked mouse thymocyte surface molecules released by phosphatidylinositol-specific phospholipase C; Pierres M et al.; We searched for mouse thymocyte surface proteins attached to the cell membrane through a phosphatidylinositol (PI)-containing glycolipid similar to that identified in the T cell-activating Thy-1 glycoprotein . Our approach was to biochemically analyse the supernatants of 125I surface-labeled thymocytes treated with 60 U/ml of Staphylococcus aureus PI-specific phospholipase C (PI-PLC) . In addition to Thy-1, two molecules of Mr 13,000 and 52,000 were found to be specifically solubilized by the enzymatic treatment . The 52,000 structure is a single basic polypeptide of Mr 50,000 under non-reducing conditions . Two-dimensional gel electrophoresis analyses resolved the 13,000 mol . wt molecules in three relatively basic components including (i) a monomeric molecule(s), a fraction of which exhibited slower migration in reducing gels, and (ii) disulfide-linked multimeric structures comprising a major component of Mr 30,000 and a minor one of Mr 45,000 . These 52,000 and 13,000 mol . wt molecules could be released from thymocytes and Escherichia coli lipopolysaccharide (LPS)-stimulated B cell blasts, but not from a variety of mature T cell populations . These data add new members to the list of PI-linked rodent lymphoid cell differentiation markers, which already includes three activation signal-transducing T cell molecules (i.e . Thy-1, Ly-6-linked T cell-activating proteins, and RT-6).

Eur J Immunol, 1987 Dec, 17(12), 1743 - 50
Recombinant human interleukin 5 is an eosinophil differentiation factor but has no activity in standard human B cell growth factor assays; Clutterbuck E et al.; Following the observation that mouse interleukin 5 (IL5) is active as a B cell growth factor (BCGF) as well as an eosinophil differentiation factor, this work was carried out to test recombinant human IL5 for BCGF activity . A highly active, partially purified batch of recombinant human IL5 was prepared and tested for BCGF activity in four laboratories . This batch gave a 50% endpoint of 1:77,450 in the human eosinophil differentiation assay, 1:983 in the mouse eosinophil differentiation assay and 1:42 in the mouse BCL1 assay, thus demonstrating that, like mouse IL5, human IL5 has cross-species activity . By comparison with the assays in the mouse this batch would be expected to have 50% maximal human BCGF activity of about 1:4000 . In each assay a known positive factor was used as a positive control, and there was no inhibitory activity in the preparation . However, despite the activity towards the mouse B cell lymphoma, the results showed no detectable activity in a panel of assays used to identify human BCGF and B cell differentiation factors . These assays included (a) proliferation assays with tonsillar or splenic B cells in the presence of the co-stimulators anti-mu or phorbol myristate acetate; (b) a restimulation assay in which tonsillar B cells are first activated with either Staphylococcus aureus Cowan 1 or a mixture of phorbol dibutyrate and ionomycin, or splenic B cells are first activated with anti-mu; (c) production of immunoglobulin by B cells in a restimulation assay with Staphylococcus aureus Cowan 1; (d) production of immunoglobulin by the Epstein-Barr virus-transformed B lymphoblastoid CESS cell line; (e) the ability to stimulate proliferation of chronic lymphocytic leukemia (B-CLL) cells freshly explanted from three different patients; (f) the ability to stimulate the B lymphoma (L4) cell line and the mature B cell (HBF1) line, and (g) the ability to replace T cells in specific antibody responses . It therefore seems unlikely that recombinant human IL5 is either a growth or a differentiation factor for human B cells, and raises the interesting question of the biological significance of the BCGF activity of this factor in the mouse.

Arch Ophthalmol, 1987 Dec, 105(12), 1699 - 702
The role of vitrectomy in the treatment of postoperative bacterial endophthalmitis . An experimental study; Talley AR et al.; While the diagnostic value of vitreous culture in the management of bacterial endophthalmitis is well established, the therapeutic value of vitrectomy in this condition is debated . The present experimental study uses an aphakic model of Staphylococcus aureus endophthalmitis in the rabbit . Animals were treated with the following: (1) intravitreal antibiotics alone; (2) intravitreal antibiotics with vitrectomy; (3) vitrectomy alone; and (4) no treatment . Eyes treated with antibiotics and vitrectomy displayed significantly clearer media at 14 days after therapy compared with eyes treated with antibiotics alone . There was also a greater tendency for eyes treated with antibiotics and vitrectomy to have negative cultures at 14 days, although this difference was not statistically significant . These findings are consistent with beneficial effects of therapeutic vitrectomy as an adjunct to intravitreal antibiotic therapy in an animal model of aphakic bacterial endophthalmitis.

J Antimicrob Chemother, 1987 Dec, 20(6), 839 - 47
Bactericidal effect of ofloxacin alone and combined with fosfomycin or vancomycin against Staphylococcus aureus in vitro and in sera from volunteers; Weber P et al.; The bactericidal activity of ofloxacin alone and in combination was evaluated against strains of Staphylococcus aureus by measuring MBCs, FBC indexes and by the killing curve technique . Bactericidal titres were determined in sera from volunteers given ofloxacin alone or in combination with fosfomycin or vancomycin . FBC indices less than 0.75 were observed with fosfomycin, showing moderate synergy . FBC indices of 1 were seen with vancomycin . Killing kinetic experiments indicated that ofloxacin (1 and 4 mg/l) exerted a rapid bactericidal effect (99.9% killing in 4 h); the combination of ofloxacin and fosfomycin was synergistic for one of three strains, while killing kinetics of ofloxacin were unaltered by fosfomycin for two of three strains or by vancomycin for the three strains . Sera collected two hours after ofloxacin or fosfomycin had been administered had bactericidal titres less than 1/2 . Bactericidal titres were significantly greater in sera from volunteers given the combination of these two drugs . Similar bactericidal titres were obtained in sera after the administration of vancomycin alone or in combination with ofloxacin . A loading dose of 400 mg ofloxacin with subsequent doses of 200 mg had no significantly prolonged effect on bactericidal titres.

Proc Natl Acad Sci U S A, 1987 Dec, 84(24), 8941 - 5
Lipophosphoglycan of Leishmania major that vaccinates against cutaneous leishmaniasis contains an alkylglycerophosphoinositol lipid anchor; McConville MJ et al.; The major cell surface glycoconjugate of Leishmania major, a putative parasite receptor for macrophages, is a lipophosphoglycan containing 81.6% (wt/wt) carbohydrate, 17.0% (wt/wt) phosphate, and 1.4% (wt/wt) lipid . It has been purified to homogeneity by hydrophobic chromatography and consists of a polydisperse family of molecules with Mr 5000-40,000 . It contains galactose, mannose, glucose, arabinose, glucosamine, and inositol in the molar ratio of 51:21:5:6:1:1 . The lipophosphoglycan has a complex structure, consisting mainly of tri- and tetrasaccharide units linked by phosphodiester bonds, which are cleaved by HF hydrolysis . The phosphate groups are located on the 6-hydroxyl of both galactose and mannose residues . The lipophosphoglycan is anchored to the parasite surface by a 1-O-alkyl-sn-glycero-3-phosphoinositol moiety . This conclusion is supported by analysis of the products of nitrous acid deamination, HF hydrolysis, and Staphylococcus aureus phosphatidylinositol specific-phospholipase C treatment . The 24:0 and 26:0 alkyl chains accounted for 93% of the ether-linked fatty acids in the lipid anchor . The results are also consistent with a glycosidic linkage between the inositol and a non-N-acetylated glucosamine residue . The lipophosphoglycan membrane anchor shares limited structural homology with the glycosylphosphatidylinositol anchors of several eukaryotic proteins, indicating that this type of membrane anchor is not limited to proteins . Vaccination of mice with the purified L . major lipophosphoglycan in liposomes induced resistance against cutaneous leishmaniasis.

Eur J Biochem, 1987 Dec 1, 169(2), 333 - 48
Panulirus interruptus hemocyanin . The elucidation of the complete amino acid sequence of subunit a; Bak HJ et al.; As a final step in the elucidation of the primary structure of subunit a of Panulirus interruptus hemocyanin (657 residues, Mr 75696 excluding two copper ions and carbohydrate), the amino acid sequence of the largest fragment obtained by limited trypsinolysis was determined . The elucidation of the sequence of residues 176-657, comprising domains two and three, was mainly based on two digests, with CNBr and trypsin, respectively, from both of which a complete set of peptides was obtained . Additional sequence information was obtained from a digest with Staphylococcus aureus V8 protease and from one fragment obtained by cleaving subunit a with hydroxylamine . A block during Edman degradations indicated an Asn-Gly sequence at positions 597-598, although only aspartic acid was identified at position 597.

Burns Incl Therm Inj, 1987 Dec, 13(6), 462 - 8
Staphylococcal toxins: screening of burn wound isolates and evidence for alpha-haemolysin production in the burn wound; Fader RC et al.; Culture filtrates of Staphylococcus aureus strains isolated from burn patients were examined for cytotoxic activities . A large molecular weight cytotoxin (MW = 253,000 daltons) that exhibited cytotoxicity for human foreskin cells and haemolytic activity against human and rabbit erythrocytes was identified . The cytotoxic activity could be completely neutralized by antiserum formed against the cytotoxin . Further characterization of the molecule by isoelectric focusing revealed that the cytotoxin was composed of at least two toxic factors of smaller molecular weight . Both factors exhibited cytotoxicity to tissue-culture cells; however, one factor lysed rabbit but not human erythrocytes whereas the other factor had the opposite haemolytic pattern . The cytotoxicity of each factor was neutralized by the antiserum formed against the cytotoxin . A cytotoxic factor that exhibited haemolytic activity for rabbit erythrocytes, and that was neutralized by the cytotoxin antiserum, was identified in burn wound extracts of mice infected with Staph . aureus . On the basis of molecular weight and isoelectric focusing data, we conclude that the large molecular weight cytotoxin was composed of an aggregation of alpha-haemolysin and another presently unidentified toxic molecule, possibly delta-toxin . Alpha-haemolysin appears to be produced in vivo during experimental staphylococcal burn wound infection.

Isr J Med Sci, 1987 Dec, 23(12), 1223 - 7
Detection of rubella-specific IgM antibodies by the staphylococcal absorption method (SAM); Barnea BS et al.; The use of SAM for the detection of rubella-specific IgM was evaluated in the sera of 318 rubella convalescents, 79 vaccinees and 53 infants with congenital rubella syndrome (CRS) . The method employed was based on the use of crude inactivated suspension of Staphylococcus aureus strain Cowan I in the hemagglutination inhibition (HI) test and 1-h incubation of antigen and antibody . Among convalescents, 99% were found positive when tested within 30 days, and 84.6% were positive within 90 days following onset of clinical symptoms . Infants with CRS were 50 to 55% positive at the age of 0 to 6 months and 23% at the age of 7 to 11 months . Overnight incubation of antigen and antibody increased the proportion of positive results, particularly among sera of rubella vaccinees . No false-positives were detected among 1,035 healthy controls; nor in 40 patients following cytomegalovirus or Epstein-Barr virus infection, nor in 22 sera containing rheumatoid factor (RF) . Comparison of SAM with a commercial enzyme-linked immunosorbent assay (ELISA) kit based on antibody capture showed 84% correlation between the two methods . Discrepancies were observed mainly in low-IgM.antibody-containing sera.

Eur J Clin Microbiol, 1987 Dec, 6(6), 679 - 81
Effect of BMY-28100, a new cephalosporin, on Staphylococcus aureus nasal carriage; Atmar RL et al.; In order to examine the in vivo activity of BMY-28100, serial quantitative nasal cultures were taken from 52 nasal carriers of Staphylococcus aureus following administration of the antimicrobial . Nasal carriage rates and mean log titers decreased significantly during (days 2-4 and 5-8) and following (days 1-2 and 6-7) treatment . No change in antibiotic sensitivities was noted in follow-up isolates, and the MIC90 remained at 2mcg/ml . BMY-28100 demonstrated good in vivo activity against Staphylococcus aureus.

Antimicrob Agents Chemother, 1987 Dec, 31(12), 1982 - 8
Effect of NaCl and nafcillin on penicillin-binding protein 2a and heterogeneous expression of methicillin resistance in Staphylococcus aureus; Chambers HF et al.; Expression of methicillin resistance in heterogeneous strains of Staphylococcus aureus is enhanced by 2 to 5% NaCl in the medium and by selection with beta-lactam antibiotics . Resistance is associated with production of a penicillin-binding protein (PBP), PBP 2a, with low affinity for binding beta-lactam antibiotics . Therefore, the effects of NaCl and nafcillin on amounts of PBP 2a produced and its binding affinity were examined and correlated with expression of resistance . Nafcillin-triggered autolysis also was examined . No relationships between the level of resistance expressed and (i) relative amounts of PBP 2a, (ii) inducibility of PBP 2a by nafcillin, or (iii) binding affinity of nafcillin for PBP 2a were found . A protective effect of NaCl for the susceptible subpopulation, corresponding to inhibition of autolysis, was observed for heterogeneous strains . Even in the absence of NaCl, highly resistant cells were relatively tolerant to nafcillin-triggered autolysis . These results support the hypothesis that high levels of resistance require an additional factor besides PBP 2a . This factor may act within the autolytic pathway.

Neuroscience, 1987 Dec, 23(3), 1143 - 55
Morphology and secretory activity of digitonin- and alpha-toxin-permeabilized chromaffin cells; Grant NJ et al.; An ultrastructural examination of cultured bovine chromaffin cells permeabilized with Staphylococcus aureus alpha-toxin or digitonin revealed differences in the preservation of cell morphology . The toxin-treated cells closely resembled control cultured cells whereas digitonin-treated cells showed gradations in cytoplasmic densities suggesting extraction, some swelling of the endoplasmic reticulum and, occasionally, discontinuities in the plasma membrane and free granules in the extracellular medium . In both cell models, there was a swelling of the mitochondria . Horseradish peroxidase labelling of permeabilized cells marked the cytoplasm of digitonin-treated cells but only the surface of toxin-treated cells, demonstrating that larger lesions were caused by digitonin . In stimulated cells, the decrease in volumetric density of chromaffin granules correlated well with catecholamine release . The sites of secretory activity could be demonstrated in toxin-treated cells using horseradish peroxidase as a surface marker . Although both cell systems secrete catecholamines in response to calcium stimulation, their calcium requirements and the kinetics of release were different . In alpha-toxin-treated cells, 100 microM free calcium induced maximal catecholamine release . In digitonin-treated cells, 20 microM evoked maximal release but secretion was blocked at 100 microM . Catecholamine release terminated in digitonin-treated cells within 10 min but continued in alpha-toxin-treated cells for at least 60 min . In addition, the maximal release observed in toxin-treated cells (50%) was always greater than that observed in digitonin-permeabilized cells (20%) . The results suggest that both exocytosis and granule translocation are operational in alpha-toxin-treated cells, but that the translocation step or the docking of granules at the plasma membrane may be impaired in digitonin-treated cells.

J Rheumatol, 1987 Dec, 14(6), 1160 - 3
Septic bursitis: presentation, treatment and prognosis; Raddatz DA et al.; Forty-nine episodes of septic bursitis in 45 patients were reviewed . Our experience concurs with previous studies: (1) the most frequently involved sites were the olecranon (63%) and prepatellar (27%) bursae; (2) Staphylococcus aureus was the commonest pathogen (78%); (3) skin breakage, trauma and/or occupational risk factors were significantly associated with infections (74 and 92% of olecranon and prepatellar episodes, respectively); (4) bursal fluid white blood cell (WBC) counts varied widely (350-392,500 WBC/mm3); and (5) a significant number of patients failed to respond to initial oral antibiotics . In addition to these points, we have been impressed with several clinical observations that merit special emphasis: (1) cellulitis adjacent to the affected bursae was frequent (89%) and often extensive; (2) profound edema occurred in 11% of affected limbs; (3) clinical resolution was slow, occurring at a mean of greater than 5 weeks, but at times requiring as long as 20 weeks to return to baseline status.

Chemioterapia, 1987 Dec, 6(6), 420 - 5
In vitro and ex vivo influence of rifamycins on human phagocytes; Bersani C et al.; We studied the effects of rifamycin SV, rifampicin and rifapentine on human phagocyte functions . Rifamycins inhibited in vitro neutrophil chemotaxis in the range of their therapeutic levels, and they significantly affected the survival of a rifamycin-sensitive strain of Staphylococcus aureus inside human monocytes . Both effects were related to the intraphagocytic penetration of these antibiotics . For the ex vivo studies, 600 mg of rifampicin were orally administered to five subjects with defective S . aureus killing . A significant reduction of neutrophil chemotaxis and increased activity against S . aureus were shown 150 and 210 min after administration of the drug.

Vaccine, 1987 Dec, 5(4), 275 - 8
Serological response of sheep to live and killed Staphylococcus aureus vaccines; Watson DL; Adult sheep were immunized intramuscularly with a killed, in vivo-grown Staphylococcus aureus vaccine combined either with dextran sulphate (group DD) or with Freund's incomplete adjuvant (group FF) . Another group (LL) received an attenuated live S . aureus vaccine intracutaneously . The animals were given a primary vaccination followed two weeks later by a booster vaccination . A fourth group of sheep (group LD) was primed with the live vaccine and given a booster vaccination with the killed vaccine combined with dextran sulphate . ELISA was used to quantify blood serum levels of IgM, IgG1 and IgG2 antibody directed against the pseudocapsular antigens of S . aureus grown under in vivo conditions . Groups LD, DD, and FF had sharp increases in mean levels of IgM antibody in the first few weeks after vaccination with another large increase in mean values for group FF at 12 weeks after primary vaccination . Group LL showed virtually no increase in levels of IgG1 antibody; the other three groups had maximum mean values for IgG1 antibody at 5 weeks (FF) and 8 weeks (LD and DD) after primary vaccination . All groups had large IgG2 antibody responses (the largest for group LD), but the response for this isotype had waned by 14 weeks after primary vaccination . Examination of the ratios of IgG2 antibody to IgG1 antibody suggested that dextran sulphate may be a useful adjuvant for preferentially stimulating synthesis of IgG2 antibody against staphylococcal pseudocapsular antigens.

J Clin Microbiol, 1987 Dec, 25(12), 2258 - 61
Collagen binding in clinical isolates of Staphylococcus aureus; Holderbaum D et al.; Collagen binding was examined in 90 strains of Staphylococcus aureus derived from patient samples . Slightly under one-half (39 of 90) of the S . aureus strains bound collagen . Collagen binding in S . aureus did not correlate with either immunoglobulin G or fibronectin binding by these organisms . Chi-square analysis of isolates obtained from patients with complicated bacteremia (bacteremia associated with deep tissue infection) compared with isolates from patients with uncomplicated bacteremia (bacteremia without other infection) showed that the former strains were significantly more likely to have collagen-binding ability . Subcloning of primary isolates from patients with bacteremia showed that all clones from individual patients were either all positive for collagen binding or all negative, suggesting a common clonal origin for this characteristic . The ability to bind collagen could not be induced in strains lacking collagen affinity by repeated subculture in media supplemented with collagen.

Ann Allergy, 1987 Dec, 59(6), 415 - 6
IgE antibodies to Staphylococcus aureus in the hypereosinophilic syndrome; Friedman SJ; A 16-year old boy with a 10-year history of circulating eosinophilia was diagnosed having the hypereosinophilic syndrome (HES) based upon the exclusion of other disorders . Eight years after the onset of his condition, he had a subcutaneous staphylococcal abscess followed by lymphangitis, rare clinical features of HES . As measured by radioallergosorbent techniques, there were significantly high serum levels of IgE antibodies to Staphylococcus aureus . The clinical significance of these antibodies is unknown, but their production may be due to persistent antigenic stimulation.

Agents Actions, 1987 Dec, 22(3-4), 288 - 94
Influence of rosmarinic acid on opsonization and intracellular killing of Escherichia coli and Staphylococcus aureus by porcine and human polymorphonuclear leucocytes; Verweij-van Vught AM et al.; The influence of rosmarinic acid on the function of porcine and human polymorphonuclear leucocytes was tested . Rosmarinic acid inhibited the chemiluminescence of PMNL, induced by preopsonized Zymosan or phorbol myristate acetate . The killing of Escherichia coli was inhibited by rosmarinic acid at a concentration of 2 mM, but not that of Staphylococcus aureus . The inhibition of the killing was due to an impaired opsonization, caused by an adverse influence of rosmarinic acid on complement activation . Direct effects of rosmarinic acid on the killing mechanisms of PMNL were not observed.

J Am Acad Dermatol, 1987 Dec, 17(6), 988 - 97
Clinical effects of diaper types on the skin of normal infants and infants with atopic dermatitis; Seymour JL et al.; Cloth diapers, cellulose core diapers (conventional disposable diapers), and cellulose core diapers containing absorbent gelling material were examined for their effects on diaper rash and skin microbiology of normal infants and infants with atopic dermatitis in a 26-week double-blind clinical trial . Infants with atopic dermatitis wearing the diapers containing absorbent gelling material had significantly lower diaper rash grades than infants with atopic dermatitis wearing cloth diapers at five of eight grading visits . Infants with atopic dermatitis wearing conventional cellulose core diapers had statistically less rash at one of eight visits . There was no statistically significant difference between diaper types at three of the eight visits . At no time did the cloth group have less diaper rash than the conventional cellulose or absorbent gelling material disposable diaper group . A statistical correlation between the severity of general atopic dermatitis outside the diaper area and the diaper rash condition under the diaper occurred only in the atopic dermatitis group wearing cloth diapers . Isolation of microorganisms from the intact, uninvolved skin surface both inside and outside the diaper showed no biologically significant changes in the presence or numbers of selected skin organisms . Repeated isolation, at multiple grading visits of Staphylococcus aureus from uncompromised skin inside the diaper area was infrequent but correlated with the diagnosis of atopic dermatitis when observed.

J Clin Pathol, 1987 Dec, 40(12), 1393 - 6
Lethal challenge of gnotobiotic weanling rats with bacterial isolates from cases of sudden infant death syndrome (SIDS); Lee S et al.; An attempt was made to produce an animal model of sudden infant death syndrome (SIDS) . The experimental animals (germ free weanling rats) were exposed to nasopharyngeal isolates from cases of SIDS to test the hypothesis that common bacteria may have an aetiological role in the disease . Negative results were obtained when the strains were tested in isolation, but certain combinations of organisms (specifically some Staphylococcus aureus and Escherichia coli) killed the animals rapidly (less than 18 hours) without prolonged terminal illness . Post mortem histological findings were consistent with those of SIDS . The lethal toxigenic potential of nasopharyngeal bacteria, which are regarded as harmless in adults, should be reconsidered in respect of the aetiology of SIDS.

Biochemistry, 1987 Dec 1, 26(24), 7744 - 8
Photoaffinity labeling of the thymidine triphosphate binding domain in Escherichia coli DNA polymerase I: identification of histidine-881 as the site of cross-linking; Pandey VN et al.; Using the technique of ultraviolet-mediated cross-linking of substrate deoxynucleoside triphosphates (dNTPs) to their acceptor site {Abraham, K . I., & Modak, M . J . (1984) Biochemistry 23, 1176-1182}, we have labeled the Klenow fragment of Escherichia coli DNA polymerase I (Pol I) with {alpha-32P}dTTP . Covalent cross-linking of {alpha-32P}dTTP to the Klenow fragment is shown to be at the substrate binding site by the following criteria: (a) the cross-linking reaction requires dTTP in its metal chelate form; (b) dTTP is readily competed out by other dNTPs as well as by substrate binding site directed reagents; (c) labeling with dTTP occurs at a single site as judged by peptide mapping . Under optimal conditions, a modification of approximately 20% of the enzyme was achieved . Following tryptic digestion of the {alpha-32P}dTTP-labeled Klenow fragment, reverse-phase high-performance liquid chromatography demonstrated that 80% of the radioactivity was contained within a single peptide . The amino acid composition and sequence of this peptide identified it as the peptide spanning amino acid residues 876-890 in the primary sequence of E . coli Pol I . Chymotrypsin and Staphylococcus aureus V8 protease digestion of the labeled tryptic peptide in each case yielded a single smaller fragment that was radioactive . Amino acid analysis and sequencing of these smaller peptides further narrowed the dTTP cross-linking site to within the region spanning residues 876-883 . We concluded that histidine-881 is the primary attachment site for dTTP in E . coli DNA Pol I, since during amino acid sequencing analysis of all three radioactive peptides loss of the histidine residue at the expected cycle is observed.

Circ Res, 1987 Dec, 61(6), 898 - 903
Myosin light chain isoforms and their phosphorylation in arterial smooth muscle; Erdodi F et al.; Arterial smooth muscle myosin contains nonphosphorylated and phosphorylated light chains that appear as 4 spots on two-dimensional, Coomassie blue-stained gel electrophoretograms at the 20,000-molecular weight level (referred to as spots 4 through 1 in order of decreasing isoelectric points) . Anti-light chain recognizes the proteins in all 4 light chain spots . Complete dephosphorylation of light chain in muscle homogenate, by inhibiting myosin light chain kinase and by adding phosphatase, leads to 2 spots on two-dimensional gel electrophoretograms; both spots are visible on immunoblots . Stimulation (K+ or stretch) of smooth muscle results in increased light chain phosphorylation . Autoradiography of the gel electrophoretograms reveals that radioactive components are contained in spots 3, 2, 1, and in an additional spot with lower isoelectric point, referred to as spot 0 . Phosphoamino acid analysis shows that spots 3 and 1 contain phosphoserine, whereas spots 2 and 0 contain phosphoserine and phosphothreonine . Two-dimensional phosphopeptide mapping of the trypsin-digested proteins from spots 3 and 1 shows predominantly 2 peptides; whereas from spots 2 and 0, it shows 5 peptides . Sodium dodecyl sulfate gel electrophoresis of the phosphopeptides obtained with Staphylococcus aureus V8 digestion gives identical maps for spots 3 and 2, which are different from the identical maps of spots 1 and 0 . The results suggest that arterial smooth muscle myosin contains 2 nonphosphorylated 20,000-dalton light chain isoforms with different amino acid sequences and that each isoform can be mono- and diphosphorylated.

J Antibiot (Tokyo), 1987 Dec, 40(12), 1716 - 32
Monocyclic beta-lactam antibiotics: synthesis and antibacterial activity of 4-(substituted ethyl)-2-azetidinone-1-sulfonic acid derivatives; Yamashita H et al.; The synthesis and antibacterial activity of sodium (3S,4R)-3-{2-(2-aminothiazol-4-yl)-(Z)-2-(O-substituted oxyimino)acetamido}-2-azetidinone-1-sulfonates having various substituted ethyl groups at the C-4 position are described . Among various substituents explored, the (substituted isothiuronio)ethyl groups were found to have strong antibacterial activity against a variety of Gram-negative bacteria, and moreover, the ethylene isothiuronium derivative exhibited moderate antibacterial activity against Staphylococcus aureus.

EMBO J, 1987 Dec 1, 6(12), 3863 - 9
Rolling circle replication of single-stranded DNA plasmid pC194; Gros MF et al.; A group of small Staphylococcus aureus/Bacillus subtilis plasmids was recently found to replicate via a circular single-stranded DNA intermediate (te Riele et al., 1986a) . We show here that a 55 bp region of one such plasmid, pC194, has origin activity when complemented in trans by the plasmid replication protein . This region contains two palindromes, 5 and 14 bp long, and a site nicked by the replication protein . DNA synthesis presumably initiated at the nick in the replication origin can be terminated at an 18 bp sequence homologous to the site of initiation, deriving from another plasmid, pUB110, or synthesized in vitro . This result suggests that, similar to the Escherichia coli single-stranded DNA phages, pC194 replicates as a rolling circle . Interestingly, there is homology between replication origins and replication proteins of pC194 and the phage phi mX174.

Eur J Immunol, 1987 Dec, 17(12), 1781 - 5
Evidence that murine hematopoietic cell subset marker J11d is attached to a glycosyl-phosphatidylinositol membrane anchor; Pierres M et al.; Glycosyl-phosphatidylinositol (G-PI) has been shown to serve as membrane anchor for cell surface molecules such as Thy-1, Ly-6-controlled ThB and Qa antigens . Here, we present several lines of evidence indicating that the hematopoietic cell lineage (i.e . thymocytes, B cell subset and red blood cells) marker defined by the rat monoclonal antibody J11d is also a G-PI-linked structure . First, surface expression of the J11d-defined molecules, and that of the related antigen B2A2, was found to be specifically reduced by treatment of thymocytes and B lymphoma or hybridoma cells with excess of Staphylococcus aureus PI-specific phospholipase C; this enzyme also solubilizes a 35-40-kDa material from erythrocyte microsomal membranes corresponding to the predominant J11d-reactive red cell surface molecules . Second, Thy-1- mutants of the BW5147, T1M1, S1A or S49 murine T lymphoma cells of the complementary classes A, B, C and E (i.e . shown to be defective in the enzymatic machinery that posttranslationally modify Thy-1 molecules) also lack J11d, or express it at a very low level . Although directed at a G-PI-linked structure, the J11d monoclonal antibody, unlike other reagents to Thy-1 or Ly-6-controlled antigens, failed to induce thymocyte proliferation even in the presence of phorbol myristate acetate and cross-linker monoclonal antibody.

Jpn J Cancer Res, 1987 Dec, 78(12), 1390 - 9
Effect of immunosuppressive factors produced by adult T cell leukemia cells on B cell responses; Tanaka Y et al.; The immunosuppressive activity of sera from adult T cell leukemia (ATL) patients and culture supernatants of ATL cells and ATL cell lines on B cell responses was examined in vitro . ATL sera and culture supernatants of ATL cells suppressed B cell proliferative and differentiative responses induced with B cell mitogens such as Staphylococcus aureus Cowan I and pokeweed mitogen . The suppressive effect was observed not only in the production of B cell growth factors (BCGF) and B cell differentiation factors (BCDF) by T cells, but also in the responsiveness of B cells to BCGF and BCDF . These results suggest that the immunosuppressive factors produced by ATL cells might act to suppress both the cellular immunity and the humoral immunity, and could have an important role in the induction of immunodeficient states in ATL patients.

J Gen Microbiol, 1987 Dec, 133 ( Pt 12), 3591 - 7
Oxygen-dependent killing of Staphylococcus aureus by human neutrophils; Edwards SW et al.; Luminol-dependent chemiluminescence was used as a monitor of reactive oxidant generation during phagocytosis of Staphylococcus aureus by human neutrophils . Reactive oxidants play a crucial role in the killing of this organism because: (a) S . aureus was killed most rapidly when the rate of increase of chemiluminescence was greatest; (b) neutrophils which had been activated to generate reactive oxidants by re-aeration of anaerobic suspensions killed this bacterium more efficiently than control suspensions; and (c) neutrophils from a patient with chronic granulomatous disease could neither generate reactive oxidants nor kill S . aureus.

Eur J Cell Biol, 1987 Dec, 45(1), 80 - 7
Interactions between protein-gold complexes and cell surfaces: a method for precise quantitation; Kehle T et al.; We have developed a rapid and precise electron microscope technique for the quantitation of gold particles in suspension using latex microspheres as a reference (EM latex technique) . This technique allowed us to determine the specific absorption of colloidal gold at its absorption maximum (520 nm) and the average number of ligands ({125I}IgG) bound to one gold particle . On the basis of these values important binding characteristics of protein-gold complexes to cell surfaces were analyzed in a model system consisting of Staphylococcus aureus with protein A on the cell wall as a specific binding site for IgG-Au . Our observations showed that the number of binding sites represented by one IgG-gold complex depended primarily on the particle size, with one 20-nm IgG-Au corresponding to 15 and one 6-nm IgG-Au to 2.5 binding sites . Hence, the efficiency of binding of IgG-Au complexes increased with decreasing gold particle size . Saturation of binding sites, however, was not achieved . The technique also made possible the determination of the affinity between IgG-Au complexes and the cell surface; this affinity can either be regarded as a characteristic of the ligand IgG or of the gold particle . We observed that the affinity of IgG decreased with the size of the gold particles to which IgG was bound, whereas the affinity of the entire gold particle increased with particle size . The EM latex technique for quantitation of gold particles extends the general use of protein-gold complexes to the quantitative characterization of their interaction with cell surface constituents.

J Antimicrob Chemother, 1987 Dec, 20(6), 849 - 55
Interaction of ceftriaxone with human polymorphonuclear neutrophil function; Labro MT et al.; Ceftriaxone, an amino-2-thiazolyl cephalosporin, has been shown to cooperate in vitro with human neutrophils for the killing of some bacteria . In this work the direct interaction with human leucocyte bactericidal function has been studied . Ceftriaxone (1000 to 1 mg/l) did not alter neutrophil chemotaxis or superoxide anion production . It also did not interfere with the chemiluminescence response of isolated PMN although a paradoxical depressive effect was observed with whole human blood in the case of zymosan stimulation . The killing of Staphylococcus aureus and Klebsiella pneumoniae was not enhanced by ceftriaxone and phagocytosis was significantly depressed only with adherent neutrophils but not when using neutrophils in liquid medium . It is concluded that the synergy observed between leucocyte and ceftriaxone for bacterial killing cannot be related to a direct stimulation of neutrophil functions and should depend on bacterial alteration.

Mol Cell Biol, 1987 Dec, 7(12), 4178 - 84
Expression of c-src in cultured human neuroblastoma and small-cell lung carcinoma cell lines correlates with neurocrine differentiation; Mellstrom K et al.; Human cell lines with neuronal and neuroendocrine features were examined for their expression of pp60c-src, the cellular homolog of the transforming gene product pp60v-src of Rous sarcoma virus . Four neuroblastoma (LA-N-5, SH-SY5Y, Paju, and SK-N-MC) and three small-cell lung carcinoma (U-2020, U-1690, and U-1285) cell lines were selected on the basis of their stage of neurocrine differentiation, as determined by the expression of neuron-specific enolase . In an immune complex protein kinase assay, all seven cell lines displayed c-src kinase activity which was considerably higher than that found in nonneurocrine cells (human diploid fibroblasts, glioma, and non-small cell lung carcinoma cell lines) . Furthermore, the c-src kinase activity, as determined by autophosphorylation or phosphorylation of an exogenous substrate, enolase, correlated with the stage of neurocrine differentiation . There was an approximately 30-fold difference in c-src kinase autophosphorylation activity between the cell lines representing the highest and lowest stages of neurocrine differentiation . A similar variation was found in the steady-state levels of the c-src protein of these cell lines . Highly differentiated neuroblastoma cells expressed two forms of the src protein . Digestion by Staphylococcus aureus V8 protease did reveal structural diversity in the amino-terminal ends of these c-src molecules . In summary, we found a clear correlation between c-src kinase activity and the stage of neuronal and neuroendocrine differentiation . Thus, the phenotypic similarity between neurons and neuroendocrine cells includes high c-src expression.

Immunology, 1987 Dec, 62(4), 581 - 6
The digestion of bacterial macromolecules by phagocytic cells: the effect of mepacrine and ethanol; Roberts PJ et al.; The digestion of radiolabelled DNA, RNA and protein from two species of bacteria was measured after their ingestion by neutrophils and monocytes . The Escherichia coli (W3110T-) strain was more susceptible to digestion than Staphylococcus aureus . Monocytes solubilized the majority of bacterial DNA to trichloroacetic acid (TCA)-soluble oligonucleotides, which were released rapidly from the cell, whereas neutrophils digested little DNA and retained most as a high molecular weight residue . Both monocytes and neutrophils released about 30% of DNA as large TCA-insoluble fragments . However, neutrophils were more proficient than monocytes at degrading bacterial RNA . Monocytes and neutrophils digested bacterial protein equally well . The drug mepacrine hydrochloride inhibited phagocytosis of bacteria by both neutrophils and monocytes, although monocytes were more sensitive to the drug . Mepacrine specifically inhibited the terminal digestion of DNA by monocytes to TCA-soluble molecules, whereas digestion to large molecular weight fragments apparently was not affected . Ethanol also inhibited the breakdown of DNA by phagocytic cells.

J Med Microbiol, 1987 Dec, 24(4), 325 - 31
Effects of mucopolysaccharides on penicillin-induced lysis of Staphylococcus aureus; Kiriyama T et al.; Effects of four mucopolysaccharides and dextran sulphate on penicillin-induced lysis of Staphylococcus aureus FDA 209P were studied . Heparin and dextran sulphate inhibited lysis, whereas hyaluronic acid enhanced it . Chondroitin sulphates A and C had no effect . Incubation of S . aureus suspended in 0.03 M phosphate buffer (pH 7.0) with dextran sulphate inhibited autolysis of the bacteria, whereas incubation with hyaluronic acid enhanced autolysis . Both extracellular and cell-associated autolysin activities of S . aureus were suppressed by dextran sulphate and high concentrations of heparin . The addition of hyaluronic acid enhanced autolysin activity . The release of lipoteichoic acid (LTA), a modulator of autolysin activity, from penicillin-treated bacteria was inhibited by heparin and dextran sulphate . However, hyaluronic acid had no effect on release of LTA . These results suggest that inhibition of penicillin-induced lysis of S . aureus by heparin results mainly from inhibition of LTA release while dextran sulphate inhibits both autolysin activity and LTA release . Hyaluronic acid appears to enhance penicillin-induced lysis through activation of the autolysins.

Virology, 1987 Dec, 161(2), 533 - 40
Monoclonal antibodies to the glycoprotein of vesicular stomatitis virus (New Jersey serotype): a method for preliminary mapping of epitopes; Bricker BJ et al.; Of the nine antigenic determinants on the glycoprotein (G) of the New Jersey serotype of vesicular stomatitis virus (VSV) identified by competitive binding of 25 monoclonal antibodies (MAbs), those relegated to epitopes I, II, III, and IV exhibited no significant ability to neutralize virus infectivity but some nonneutralizing MAbs cross-reacted by ELISA with the G protein of VSV-Indiana . High-titered neutralization of homotypic virus was exhibited by epitope V, VI, and VII MAbs but quite variable neutralizing activity was found among MAbs of epitope "family" VIII and particularly the heterogeneous epitope "family" IX . Peptide mapping of the epitopes was not feasible because most MAbs would not bind by Western blotting to G protein under standard conditions of proteolysis or disulfide bond reduction . Therefore, a technique was devised for roughly locating epitopes by protease footprinting of G protein partially protected by individual MAbs complexed with staphylococcal protein A-Sepharose beads . Under these conditions, MAbs to all nine epitopes protected a similar 12-kDa fragment of the G protein from proteolysis by Staphylococcus aureus V8 protease . N-Terminal amino acid sequencing mapped two of these 12-kDa peptide footprints to a position on the G protein extending from amino acid 219 to about 100 amino acids downstream . Although MAbs to only one epitope bound to the 12-kDa fragment by Western blotting, these data suggest, but do not prove, that all nine epitopes of the undenatured VSV-New Jersey G protein are clustered at the middle 20% of a highly structured protein . This method may help to identify the general regions for epitopes on complex proteins of as yet unknown three-dimensional structure.

FEBS Lett, 1987 Nov 16, 224(1), 128 - 32
Primary structure of ovine pituitary basic fibroblast growth factor; Simpson RJ et al.; The complete amino acid sequence of basic FGF (146 residues) from ovine pituitary glands has been established . This has been achieved by the sequence analysis of subnanomole amounts of the intact molecule and of peptides derived by enzymatic digestions with clostripain, chymotrypsin, pepsin and Staphylococcus aureus V8 protease . Microbore HPLC, employing 1-2 mm i.d . columns, was used to purify, concentrate and buffer-exchange the FGF peptides . A novel application of ion-pairing chromatography was employed to isolate peptides which were not retained on conventional reversed-phase systems . There is only one positional difference between the ovine and bovine basic FGFs, but there are 3 positional differences between ovine and human basic FGFS.

Cancer Res, 1987 Nov 15, 47(22), 5954 - 9
Antibody-directed targeting of liposomes to human cell lines: role of binding and internalization on growth inhibition; Berinstein N et al.; Small unilamellar liposomes containing methotrexate or methotrexate-gamma-aspartate were conjugated to Staphylococcus aureus protein A and were thus able to bind cell-specific immunoglobulins for targeting to malignant human B- and T-cell lines . We were able to demonstrate enhanced protein A liposome uptake and growth inhibition by targeting with an anti-major histocompatibility complex class II antibody recognizing two different B-cell lines . The enhanced growth inhibition was specific for the targeting antibody and amounted to a 2- to 3-fold lowering of the concentration of drug required to inhibit cell growth by 50% as compared to nontargeted liposomes or liposomes targeted with an antibody not recognizing a cell surface antigen . A strong association between enhanced growth inhibition and liposome internalization as assessed by fluorescent-activated cell sorter analysis of carboxyfluorescein containing protein A liposomes was seen . By contrast, specific enhancement of growth inhibition was not seen with several anti-idiotype antibodies or antibodies to T-cell differentiation antigens . Liposome internalization did not occur with these antibodies . Failure of growth inhibition and PA liposome internalization could not be explained by differences in cell binding of the antibody PA liposomes or the degree of protein A binding of the targeting antibody . Although the ability of the targeting antibody to bind to the cell and to protein A are important, these factors alone are not sufficient to guarantee internalization and growth inhibition . Variations in rates of internalization of various cell surface antigen-antibody complexes may account for different protein A liposome mediated cytotoxicities.

J Biol Chem, 1987 Nov 15, 262(32), 15538 - 44
Autophosphorylation of the protein kinase dependent on double-stranded RNA; Galabru J et al.; The double-stranded RNA (dsRNA)-dependent protein kinase (p68 kinase) from interferon-treated human cell is a Mr 68,000 protein induced by interferon . By the use of a specific monoclonal antibody, we have been able to study the two distinct protein kinase activities characteristic of purified p68 kinase . The first activity is functional for endogenous phosphorylation of the enzyme (p68 kinase), whereas the second one is responsible for the phosphorylation of exogenous substrates such as eukaryotic initiation factor 2 and histone . When activated by dsRNA in the presence of Mn2+ and ATP, p68 kinase is autophosphorylated and is then capable of catalyzing phosphorylation of histone in the absence of dsRNA . Whereas binding of 8-azido-{alpha-32P} ATP (8-N3ATP) to p68 kinase is dependent on both dsRNA and Mn2+, phosphorylated p68 kinase binds 8-N3ATP independent of dsRNA . This is consistent with a dsRNA requirement for the autophosphorylation of p68 kinase, but not for the phosphorylation of exogenous substrates . p68 kinase is mainly associated with the ribosomal pellet . It could be recovered efficiently by a buffer containing both high salt and a nonionic detergent . Synthesis of p68 kinase is induced several-fold by interferon in different types of human cells . Partial proteolysis of {35S}methionine and an 8-N3ATP-labeled p68 kinase preparation by Staphylococcus aureus V8 protease indicated the presence of a major Mr 48,000 polypeptide (p48) with a specific ATP-binding site . p48 probably contains the catalytic unit of p68 kinase and is analogous to a similar protein which we have previously described as a distinct protein present in a complexed form with p68 kinase . We now believe that the presence of p48 in previously purified kinase preparations was due to partial degradation of p68 kinase.

J Biol Chem, 1987 Nov 15, 262(32), 15746 - 51
Characterization of transducin from bovine retinal rod outer segments . Participation of the amino-terminal region of T alpha in subunit interaction; Navon SE et al.; The GTP-induced dissociation of T alpha from T beta gamma initiates the release of transducin from photolyzed rhodopsin and the subsequent activation of the cGMP phosphodiesterase . In this study, site-specific proteolysis and immunoprecipitation were used to map the domain of T alpha that interacts with T beta gamma . We found that Staphylococcus aureus V8 protease rapidly removes a small fragment from T alpha under native conditions, resulting in the formation of a single 38-kDa polypeptide (T alpha') . Under the same conditions, T beta gamma remains intact . A 4.5-fold decrease in the rate of T alpha cleavage by S . aureus protease was observed in the presence of T beta gamma, suggesting T beta gamma binding blocks the protease-sensitive site on T alpha . Amino acid sequence analysis indicated that T alpha' is derived from the cleavage of T alpha at Glu-21 . The ability of T alpha' to interact with and activate the retinal phosphodiesterase is not diminished . However, T alpha' is unable to participate in T beta gamma-dependent activities such as the light-stimulated binding of guanine nucleotides, binding to photoexcited rhodopsin, and ADP-ribosylation catalyzed by pertussis toxin . Moreover, the anti-T alpha monoclonal antibody TF16 was able to precipitate T beta gamma in the presence of T alpha, but not with either T alpha' or T alpha-guanosine 5'-O-(3-thiotriphosphate) . We conclude that the amino-terminal region of T alpha participates in T beta gamma interaction and discuss our results with respect to the known structure and function of transducin.

J Biol Chem, 1987 Nov 5, 262(31), 15285 - 90
A phospho-oligosaccharide mimics the effect of insulin to inhibit isoproterenol-dependent phosphorylation of phospholipid methyltransferase in isolated adipocytes; Kelly KL et al.; Addition of isoproterenol to isolated rat adipocytes prelabeled with {32P}phosphate caused an increase in the phosphorylation and activation of phospholipid methyltransferase . 32P-Labeled phospholipid methyltransferase was recovered by immunoprecipitation and gel electrophoresis . Analysis of 32P-labeled peptides revealed one site of phosphorylation regulated by isoproterenol, and analysis of phosphoamino acids demonstrated that the incorporation of {32P}phosphate was on phosphoserine . Incubation of adipocytes with isoproterenol in the presence of insulin or a phospho-oligosaccharide inhibited the phosphorylation and activation of this enzyme . The inhibitory effect of insulin on the phosphorylation of phospholipid methyltransferase was reversible, and it was mimicked by a phospho-oligosaccharide . The phospho-oligosaccharide was generated by hydrolysis of an isolated glycophospholipid with phosphatidylinositol-specific phospholipase C from Staphylococcus aureus . The insulin-like effect of this phospho-oligosaccharide on the phosphorylation of phospholipid methyltransferase was demonstrated in isolated adipocytes, and the effect was abolished by treatment of the phospho-oligosaccharide with 10% NH4OH, nitrous acid, or sodium periodate . These data suggest that in intact adipocytes the effect of insulin to inhibit the phosphorylation/activation of phospholipid methyltransferase is mediated by a phospho-oligosaccharide generated by a phosphatidylinositol-specific phospholipase C.

Biochemistry, 1987 Nov 3, 26(22), 7090 - 102
The 43-kilodalton protein of Torpedo nicotinic postsynaptic membranes: purification and determination of primary structure; Carr C et al.; The primary structure of the 43-kilodalton peripheral membrane protein (43-kDa protein) of Torpedo nicotinic postsynaptic membrane has been determined . The 43-kDa protein, which was isolated by preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis, has an amino terminus resistant to Edman degradation, while the sequence at the carboxyl terminus is Tyr-Val . An amino acid sequence of 405 residues was obtained by NH2-terminal sequence analysis of complementary peptides generated by digestion with trypsin, chymotrypsin, Staphylococcus aureus V8 protease, and endoproteinase Lys-C, as well as by chemical cleavage at methionine . This sequence of molecular mass 45,618 daltons lacks the amino terminus but extends to the carboxyl terminus of the 43-kDa protein . Unusual structural features of the 43-kDa protein include two regions of approximately 80 residues, each containing 10% cysteine, as well as stretches predicted to exist as amphipathic alpha-helices . Other than the group blocking the amino terminus, no evidence was found for posttranslational modification of amino acids . The 43-kDa protein may represent a novel protein family because a computer search of this sequence with the National Biomedical Research Foundation data base (Release 12.0) did not reveal any significant homology to known protein sequences.

Neurosurgery, 1987 Nov, 21(5), 744 - 7
Epidural lipomatosis simulating an epidural abscess: case report and literature review; Haid RW Jr et al.; A case of epidural lipomatosis in a 49-year-old man presenting with paraparesis, midthoracic pain, and Staphylococcus aureus pneumonia is reported . The patient had been on low dose corticosteroid therapy for 7 years for rheumatoid arthritis . The clinical and myelographic findings suggested a diagnosis of epidural abscess, but the only abnormality discovered at operation was abundant fatty tissue in the dorsal epidural space significantly compressing the spinal cord, and this was partially removed . Postoperative neurological improvement suggested that the lipomatosis was responsible for the spinal cord compression and dysfunction . If this diagnosis had been suspected, it might have been confirmed by magnetic resonance imaging or postmyelography computed tomographic scanning . With such a diagnosis, an alternative treatment could have been to decrease the steroid dose, observe for clinical improvement, and perhaps avoid operation.

Stroke, 1987 Nov-Dec, 18(6), 1048 - 56
Mechanisms of intracranial hemorrhage in infective endocarditis; Hart RG et al.; Analysis of 17 patients with infective endocarditis and intracranial hemorrhage yielded several different mechanisms of bleeding . Nine of 15 (60%) symptomatic intracranial hemorrhages occurred within 48 hours of admission and 3 more (20%) after hospital discharge . In 7 patients with Staphylococcus aureus endocarditis, symptomatic intracranial hemorrhage occurred within 48 hours of admission and resulted from septic arteritis in all 3 examined pathologically . Secondary hemorrhagic transformation (hemorrhagic infarction) was asymptomatic in 2 nonanticoagulated patients but was associated with clinical worsening in 2 anticoagulated patients . Anticoagulation potentially contributed to intracranial hemorrhage in 4 of the 17 patients (24%) . Proven mycotic aneurysms were present in only 2 patients (12%), 1 of whom presented with massive, fatal intracranial hemorrhage . Mycotic aneurysms amenable to surgery are uncommon and underlie only a fraction of intracranial hemorrhages in infective endocarditis.

J Allergy Clin Immunol, 1987 Nov, 80(5), 746 - 51
Evolution of the hyperimmunoglobulin E and recurrent infection (HIE, JOB's) syndrome in a young girl; Dreskin SC et al.; The hyperimmunoglobulin E and recurrent infection syndrome is difficult to diagnose in children with markedly elevated IgE and recurrent superficial Staphylococcus aureus infections who have not presented with a severe infection . The patient, the child of a woman with HIE, had elevated cord blood IgE . In early infancy, she had cutaneous colonization with S . aureus followed by frank impetiginous lesions . Anti-S . aureus IgE was easily detected with a highly specific ELISA assay at 2 years of age (2 years before her presentation with a S . aureus subcutaneous abscess) . Thus, the measurement of anti-S . aureus IgE by this technique may be a useful laboratory test for the diagnosis of HIE before the appearance of a severe infection.

Ann Thorac Surg, 1987 Nov, 44(5), 523 - 8
Bubble oxygenation and cardiotomy suction impair the host defense during cardiopulmonary bypass: a study in dogs; van Oeveren W et al.; Decreased complement levels and impairment of polymorphonuclear leukocyte function increase the risk of infection during cardiopulmonary bypass (CPB) . The effects of different types of oxygenator and of blood suction on this natural humoral and cellular host defense mechanism were investigated in dogs undergoing CPB during sham open-heart operations . Airborne contamination of the wound area and the CPB circuit was performed by aerosolizing Staphylococcus aureus . A membrane oxygenator in the CPB circuit maintained a normal host defense mechanism . The use of cardiotomy suction during CPB with this type of oxygenator affected the host defense to some extent . The use of a bubble oxygenator in the CPB circuit together with cardiotomy suction seriously impaired the host defense . Postoperatively bacteremia developed in no dogs in the membrane oxygenator group, whereas 8 of 15 dogs in the bubble oxygenator group had a positive blood culture for the indicator microorganism . We conclude that the use of a membrane oxygenator is helpful to maintain the host defense . Attention has to be paid to reduce the deleterious effects of cardiotomy suction.

Ann Surg, 1987 Nov, 206(5), 666 - 73
PTFE grafts for hemodialysis access . Techniques for insertion and management of complications; Raju S; In a series of 602 procedures, over 90% of primary forearm insertions of PTFE grafts between the radial artery and a cubital vein were possible . Thrombosis of the graft, which was invariably due to venous outflow obstruction, was the most common complication encountered . Revision of the venous anastomosis was not necessary in about one-third of the thrombosed grafts if a size 3 coronary dilator could be passed and the augmentation test was satisfactory . For revisions, creation of a new venous anastomosis using a jump graft was preferred over patch angioplasty or venous endarterectomy . Crossing the elbow for this purpose did not adversely affect graft patency . The incidence of aneurysm formation and infection was 16% and 35%, respectively . Infections involving the graft were managed by drainage, antibiotics, and bypass of the infected portion . Immediate bypass and delayed bypass were equally effective . About one-half of the infected grafts were salvaged by these techniques . The most common organism was Staphylococcus aureus . With a combination of the techniques outlined above, the service life of individual PTFE grafts can be extended . Two-year access patency in this series was 77%.

Am J Pathol, 1987 Nov, 129(2), 217 - 22
Pentoxifylline enhancement of defective neutrophil function and host defense in neonatal mice; Krause PJ et al.; Decreased neutrophil (PMN) chemotaxis is thought to contribute to the increased morbidity and mortality from infection in newborn infants . Pentoxifylline, a methylxanthine, has previously been shown to augment PMN chemotaxis in vitro . The authors therefore investigated the effects of pentoxifylline on 1) in vitro PMN chemotaxis, 2) in vivo leukocyte accumulation, and 3) protection against Staphylococcus aureus infection in newborn mice . Using a modified Boyden chamber system, they demonstrated that pentoxifylline significantly enhanced neonatal PMN chemotaxis in a dose-dependent manner . Additionally, pentoxifylline was found to increase PMN accumulation in vivo in a proteose peptone-induced peritonitis model . Finally, the survival rate in experimentally induced S aureus infection was 51% in neonatal mice given pentoxifylline, compared with 17% in a control (nonpentoxifylline) group (P less than 0.01) . These data demonstrate pentoxifylline modulation of PMN migration and enhancement of host defense against bacterial infection.

J Med Microbiol, 1987 Nov, 24(3), 275 - 81
Esterase electrophoretic polymorphism of methicillin-sensitive and methicillin-resistant strains of Staphylococcus aureus; Branger C et al.; Methicillin-sensitive and methicillin-resistant strains of Staphylococcus aureus from diverse geographic origins were analysed by polyacrylamide-agarose gel electrophoresis for esterase polymorphism . Three kinds of esterase bands, designated A, B and C, were defined by their ranges of activity toward five synthetic substrates and their resistance to di-isopropyl fluorophosphate . There were five allozymes of esterase A, four of esterase B and four of esterase C . Eighteen distinct combinations of allozymes (zymotypes) were distinguished amongst 105 strains analysed . Two major zymotypes were represented by 35 and 19 strains respectively, whereas other zymotypes were represented by one or, at most, seven strains . The coefficient of genetic diversity was lower for methicillin-resistant strains than for methicillin-sensitive strains . Most of the methicillin-resistant strains are represented by the two major zymotypes which differed from each other by the electrophoretic behaviour of the three esterases . These results indicate that, on the basis of esterase electrophoretic polymorphism, methicillin resistance is expressed in genetically different strains.

Infect Immun, 1987 Nov, 55(11), 2747 - 53
Intrapulmonary growth of Staphylococcus aureus in rats during induced atelectasis; Frederick D et al.; Intrinsic pulmonary antibacterial defenses are mediated by alveolar macrophages and by noncellular factors . Mechanical ventilation in the resting tidal volume range leads to alterations in the physical characteristics of alveolar surfactant, alveolar instability, regional hypoxia, and systemic hypoxemia . While a number of experimental manipulations diminish the activity of the intrinsic antibacterial defense system, the effects of mechanical ventilation per se have not been systematically evaluated previously . We found that normal rats ventilated without sighing (periodic large breaths) manifested severe defects in pulmonary clearance of Staphylococcus aureus during 6-h experiments, such that growth of the inoculum occurred . Addition of a timer-controlled mechanism to cause the animals to sigh every 2 min, without other modifications in the experimental conditions, caused significant improvement in clearance . Analysis of cellular response, compartmentalization of viable bacteria, surfactant quantities and sedimentation characteristics, and protein influx indicated that the defect in clearance paralleled alterations in the physical state of surfactant and alveolar stability but was not strongly correlated with alterations in the other parameters we measured . The data show that defective pulmonary bacterial clearance is rapidly induced by measures which alter alveolar stability and suggest that intrinsic pulmonary defenses require maintenance of normal air-liquid interfaces for optimal function.

Clin Orthop, 1987 Nov, (224), 33 - 6
Infection in experimental arthroplasties; Southwood RT et al.; Experimental bacterial infection following implant arthroplasties was investigated in rabbits . Bone cement and a stainless steel head and stem prosthesis were inserted after reaming of the femoral neck and shaft . Measured doses of Staphylococcus aureus were injected either into the femoral medullary cavity or intravenously . Intravenous challenge required high inocula to establish arthroplasty infection, whereas infection around the prosthesis was consistently established after inoculation of 10(3) bacteria into the femoral medulla . The erythrocyte sedimentation rate (ESR) proved the most reliable clinical test of infection . Hematogenous infection was difficult to reproduce . Three weeks after operation, the arthroplasty was as resistant as the normal hip . Antibiotics were administered in doses equivalent to doses used in the treatment of infections in humans . When gentamicin-impregnated cement was used, 60 times the number of organisms were required to establish infection . Flucloxacillin and Imipenem gave similar protection . Rifampicin gave protection to a level exceeding the lethal dose of organisms for the model.

Pediatr Pulmonol, 1987 Nov-Dec, 3(6), 425 - 8
Diagnostic bronchoalveolar lavage in children with AIDS; Bye MR et al.; Between October, 1985 and May 1987, 29 children (mean age 22 +/- 22 months, range 2-54 months) with AIDS or ARC developed acute respiratory illness . The initial diagnostic procedure was flexible fiberoptic bronchoscopy, with bronchoalveolar lavage (BAL) . BAL was positive for Pneumocystis carinii in 14 and for respiratory syncytial virus, Staphylococcus aureus, and Escherichia coli in 3 additional patients . Subsequent lung tissue analysis and/or clinical course suggested no false negative lavages . Complications possibly related to the procedure occurred in two patients . We find BAL an effective diagnostic technique in these patients, offering a less invasive alternative to open lung biopsy.

J Immunol, 1987 Nov 1, 139(9), 2925 - 8
Human monoclonal IgG rheumatoid factor has structural homology with bacterial Fc receptor proteins; Weisbart RH et al.; A cloned lymphoblast cell line, hRF-1, that secreted human monoclonal IgG4 rheumatoid factor autoantibody was produced by Epstein-Barr virus transformation of lymphocytes from rheumatoid arthritis synovium . The binding of hRF-1 rheumatoid factor to IgG globulins of different mammalian species was similar to the binding specificity of Staphylococcus aureus protein A (SpA) and to antibodies found in the sera from patients with rheumatoid arthritis . hRF-1 also had the same binding pattern to human IgG subclasses as SpA . Direct competition was observed between SpA and hRF-1 in binding IgG Fc . These results provide evidence for structural homology between a bacterial Fc receptor protein (SpA) and the monoclonal IgG rheumatoid factor.

J Clin Endocrinol Metab, 1987 Nov, 65(5), 853 - 61
Induction of thyroid autoantibody production: synergistic effect of B cell mitogen combined with T cell mitogen; Iwatani Y et al.; In vitro production of thyroglobulin autoantibodies (TgAb) and thyroid microsomal autoantibodies (McAb) by peripheral blood mononuclear cells (PBMC) stimulated with the B cell mitogen Staphylococcus aureus Cowan I (SAC) and the T cell mitogen pokeweed mitogen (PWM) was examined in 35 normal subjects (NC) and 64 patients with autoimmune thyroid disease (AITD) using an enzyme-linked immunosorbent assay technique . Low concentrations of SAC plus PWM resulted in a synergistic effect on thyroid autoantibody production as well as nonspecific immunoglobulin G production . With such maximal stimulation, TgAb production was detected in all PBMC preparations from serum TgAb-positive patients with AITD; TgAb production was also detected in some NC (46%) and serum TgAb-negative patients with AITD (39%), but the levels of TgAb production were low . Similarly, McAb production was marked in PBMC preparations from serum TgAb-negative but McAb-positive patients . TgAb-secreting cells were also detected in NC by the plaque-forming cell (PFC) assay . The response patterns of PBMC to mitogen (Nil, PWM, and SAC plus PWM) in terms of TgAb production varied among serum TgAb-positive patients with AITD, but not among NC and serum TgAb-negative patients with AITD . Serum TgAb titers were significantly correlated with the in vitro production of TgAb by PBMC with no stimulation (r = 0.64; n = 99; P less than 0.001), with stimulation by PWM (r = 0.75), and with stimulation by SAC plus PWM (r = 0.87); the correlation coefficient increased with the efficiency of stimulation of B cell differentiation . Similar results were found for McAb production . These data suggest that 1) optimal in vitro thyroid autoantibody production occurs with B cell mitogen (SAC) acting synergistically with T cell mitogen (PWM); 2) sufficient numbers of resting B lymphocytes specific for Tg or microsomal antigens are present in some NC PBMC; 3) stages of thyroid-specific B cell differentiation in PBMC vary among serum thyroid autoantibody-positive patients with AITD; and 4) the potential of PBMC to produce thyroid autoantibodies may correlate with the capacity of thyroid-derived lymphocytes . Thus, the circulating lymphocytes may provide a useful vehicle by which sequential changes occurring at the tissue level may be examined.

J Biochem (Tokyo), 1987 Nov, 102(5), 1177 - 86
Nucleotide sequence of the staphylocoagulase gene: its unique COOH-terminal 8 tandem repeats; Kaida S et al.; The entire staphylocoagulase gene of Staphylococcus aureus strain BB was cloned on a MboI restriction endonuclease fragment inserted into pAT153 plasmid vector . The staphylocoagulase was expressed in Escherichia coli, as judged by the formation of a fibrin halo on an agar plate containing rabbit plasma and bovine fibrinogen . We have determined the complete nucleotide sequence of the staphylocoagulase gene by the dideoxynucleotide chain termination method . The deduced amino acid sequence consisted of 715 residues including a signal peptide of 26 residues . Therefore, the predicted molecular weight of the mature protein was 77,337 . This sequence was corroborated by reference to the amino acid compositions of 30 lysyl endopeptidase peptides and the sequences of 12 of these peptides isolated from the purified staphylocoagulase . The 5'-flanking region was found to contain a putative Shine-Dalgarno sequence and a putative "-10" element for transcription . The COOH-terminal stretch of 216 amino acids of staphylocoagulase was composed of 8 tandem repeats each consisting of 27 amino acid residues . The amino acid sequence of staphylocoagulase derived from strain BB showed 57% identity with that of the chymotryptic 43-kDa fragment of staphylocoagulase isolated previously from strain 213 (Kawabata, S., Miyata, To., Morita, T., Miyata, Ta., Iwanaga, S., & Igarashi, H . (1986) J . Biol . Chem . 261, 527-531).

Jpn J Antibiot, 1987 Nov, 40(11), 1941 - 5
{Antibacterial activities of various aminoglycosides against clinically isolated methicillin-resistant Staphylococcus aureus}; Deguchi K et al.; Methicillin-resistant Staphylococcus aureus (MRSA) were isolated from samples collected from various patients during 1986, and antibacterial activities of 6 aminoglycosides (AGs) (netilmicin (NTL), gentamicin (GM), sisomicin (SISO), dibekacin (DKB), tobramycin (TOB) and amikacin (AMK} and 4 beta-lactam antibiotics (cefazolin (CEZ), cefmetazole (CMZ), cloxacillin (MCIPC) and methicillin (DMPPC) against these MRSA were evaluated . Among these 6 AGs, NTL was the most potent, and its MIC50 and MIC80 were 1.56 and 3.13 micrograms/ml, respectively . Antibacterial activities of GM, SISO, DKB and TOB were weak, and MIC50's of GM and DKB were both 100 micrograms/ml, while those of SISO and TOB were 50 and greater than 100 micrograms/ml, respectively . Frequency of highly resistant specimens to AMK was rather low and its MIC50 and MIC80 were 12.5 and 25 micrograms/ml, respectively . As for antibacterial activities of the above 4 beta-lactam antibiotics, the MIC50 and MIC80 of CMZ were 6.25 and 12.5 micrograms/ml, respectively, and therefore, its antibacterial activity to MRSA is relatively good . However, MIC50's of CEZ, MCIPC and DMPPC were all greater than 100 micrograms/ml, showing poor antibacterial activities . Recently, MRSA became a problem in various fields of clinical practice, and a number of literatures reporting refractory infections caused by MRSA have been published . Since MRSA is featured as multiply resistant bacteria, it is known that MRSA is resistant to the majority of existing antibiotics (penicillins, cephems, macrolides, AGs, etc.) . In 1985, we reported results of our study concerning the antibacterial activities of a number of CEPs and some of AGs against multiply resistant S . aureus including MRSA.(ABSTRACT TRUNCATED AT 250 WORDS)

J Appl Bacteriol, 1987 Nov, 63(5), 417 - 25
Use of plasmid profiles to detect changes in strains of Staphylococcus aureus during poultry processing; Dodd CE et al.; The plasmid profiles of Staphylococcus aureus strains isolated at different stages in three poultry processing plants have been examined . Changes in profiles were seen in two plants after the plucking stage and the appearance of these new profiles correlated with the presence of an endemic strain, as suggested previously by increases in bacterial counts and changes in biotypes at the same stage . A third plant in which such changes did not occur showed no change in profiles . Plasmid profiles are therefore a rapid and sensitive method for distinguishing endemic strains within a plant from the flora of the incoming birds . Certain profiles also appeared to correspond with particular biotypes and certain phage types.

Diagn Microbiol Infect Dis, 1987 Nov, 8(3), 137 - 47
In vitro activity of novobiocin and rifampin alone and in combination against oxacillin-resistant Staphylococcus aureus; Johnston BL et al.; Rifampin and novobiocin both have excellent activity against oxacillin-resistant Staphylococcus aureus, but their single use may be associated with the development of resistance . To help predict their clinical value in our institution, 60 recent clinical isolates of oxacillin-resistant S . aureus were studied for in vitro susceptibility to the two agents . Ten isolates with increased MICs to both agents or to rifampin alone were also studied by modified checkerboard and kill-curve methods . Indifference was consistently demonstrated by the checkerboard method and generally found in kill-curve studies . Prevention of development of resistance was demonstrated with the antimicrobial combinations for some isolates . Isolates with increased MICs for the two agents fell into two distinctive groups, with prevention of the development of rifampin resistance occurring in one group but not in the other, suggesting that different strains of oxacillin-resistant S . aureus may have different capacities for development of rifampin resistance.

Antibiot Med Biotekhnol, 1987 Nov, 32(11), 865 - 8
{Anaerobic microflora of patients with suppurative and septic complications after non-hospital abortions}; Miasnikova LG et al.; Microflora of pathological biosubstrates from 25 patients aged from 18 to 41 years with criminal abortion complications such as sepsis, septic shock, septicemia, and septic pyemia, peritonitis and endometritis of various severity was studied . Obligate anaerobic organisms in association with facultative anaerobes were detected in 84 per cent of the patients . Bacteroids were isolated from operation materials of 36 per cent of the patients . Bacteroids in association with Staphylococcus aureus, peptostreptococci and enterococci were recorded in 16, 8 and 24 per cent of the patients, respectively . Composition of the anaerobic and facultative anaerobic microflora was analyzed in the patients with local and general infections . Antibiotic sensitivity assay of the bacteroids showed that rifampicin, metronidazole, levomycetin (chloramphenicol) and clindamycin were the most active drugs . The use of anaerobic techniques enabled to demonstrate that in patients with purulent septic complications of criminal abortion there prevailed anaerobic-aerobic associations . The results should be considered in treatment of gynecological patients with purulent septic infections.

Infection, 1987 Nov-Dec, 15(6), 459 - 62
Plasmid fingerprinting of methicillin-resistant Staphylococcus aureus strains isolated in Hamburg; Gatermann S; By means of restriction endonuclease digests and DNA/hybridisation studies we analysed ten representative methicillin-resistant Staphylococcus aureus strains of our collection for plasmid similarities and plasmid associated resistance determinants . We found that strains isolated at our laboratory contained identical or at least most similar plasmids . Isolates from another geographical origin showed different plasmid patterns . We found resistance determinants for gentamicin to be chromosomally encoded, whereas resistance to heavy metal ions and chloramphenicol was always plasmid associated . Resistance to trimethoprim, tetracycline and erythromycin was usually chromosomally mediated but could also reside on a plasmid . Our results indicate that methicillin-resistant strains from our collection may have a common origin . The clinical relevance of these results is discussed.

Antimicrob Agents Chemother, 1987 Nov, 31(11), 1727 - 33
Effects of temperature, NaCl, and methicillin on penicillin-binding proteins, growth, peptidoglycan synthesis, and autolysis in methicillin-resistant Staphylococcus aureus; Madiraju MV et al.; Methicillin-resistant Staphylococcus aureus strains produce a fifth penicillin-binding protein (PBP), PBP 2', with low affinity for beta-lactam antibiotics that is believed to represent a beta-lactam-insensitive peptidoglycan transpeptidase . In an effort to evaluate the adequacy of PBP 2' as an explanation of methicillin resistance, PBP 2' production and the responses of growth and peptidoglycan synthesis to methicillin under different environmental conditions have been compared . In the heterogeneous methicillin-resistant strain DU4916-K7, less PBP 2' was produced at 40 degrees C than at 30 degrees C, but inclusion of 5% (wt/vol) NaCl in the medium at 40 degrees C boosted PBP 2' production and allowed growth of the organism in the presence of 10 micrograms of methicillin per ml . When exponential-phase cultures were challenged with methicillin, growth and peptidoglycan synthesis were much more resistant at 30 degrees C than at 40 degrees C . Inclusion of NaCl in medium rendered growth and peptidoglycan synthesis more methicillin resistant at 40 degrees C . Hence, there was a good correlation between PBP 2' production and methicillin-resistant peptidoglycan synthesis under these conditions . However, PBP 2' production was increased by NaCl at 30 degrees C without markedly affecting the susceptibilities of growth and peptidoglycan synthesis to methicillin . Pregrowth of cells with methicillin, which was expected to boost PBP 2' production, seemed to increase the susceptibilities of growth and peptidoglycan synthesis to methicillin . Patterns of growth and peptidoglycan synthesis susceptibilities to methicillin which were similar to those described above were found in chloramphenicol-inhibited cultures, in which presumably no induction of PBP 2' could occur during the methicillin challenge period . Complex effects were noted in the combination of subinhibitory methicillin and NaCl . Growth of cells in the presence of NaCl stimulated their autolytic activity, which was further increased by growth with subinhibitory methicillin in addition to NaCl . It appears that NaCl enhances methicillin resistance by stimulating PBP 2' production and providing osmotic support but opposes it by stimulating autolytic activity which is exacerbated by the very low cross-linking of peptidoglycan in methicillin-resistant strains grown in the presence of methicillin.

Am J Vet Res, 1987 Nov, 48(11), 1638 - 41
Effect of estradiol and progesterone on antistaphylococcal activity of neutrophils from ovariectomized mares; Strzemienski PJ et al.; Neutrophils isolated from jugular blood of ovariectomized mares were studied for the effect of estradiol and progesterone on bactericidal activity against Staphylococcus aureus . In experiment 1, neutrophils obtained from 4 mares were tested for bactericidal activity by adding estradiol (43 pg/ml) or progesterone (6.4 ng/ml) to the bactericidal assay . In experiment 2, 3 of the 4 ovariectomized mares were given 2 mg of estradiol, IM, daily for 3 days . Eighteen days after the initial estradiol injection, mares were given 300 mg of progesterone, IM, for 6 days . Neutrophils from these mares were tested for bactericidal activity 4 days after the initial estradiol injection, 17 days after the initial estradiol injection (control), and 7 days after the first progesterone injection . Bactericidal activity was measured at 30 and 120 minutes by counting the number of colony-forming units remaining . Neutrophil antistaphylococcal activity was not altered by adding estradiol and progesterone to the assay or by supplementing ovariectomized mares with estradiol and progesterone (P greater than 0.05).

Ukr Biokhim Zh, 1987 Nov-Dec, 59(6), 50 - 4
{Formation of the transmembrane electrochemical potential on Staphylococcus cells and membrane vesicles}; Vinnikov AI; The generation of transmembrane difference of electrochemical potentials was registered on the intact cells and ultrasonication-obtained membrane vesicles of Staphylococcus aureus with the application of transmembrane electrophoresis of permeant anions, potassium transport in the presence of valinomycin and 8-anilinonaphthalene-1-sulphonate fluorescence . The membrane potential is formed when the chain of electron transfer or H+-ATPase functions or when the pH gradient varies (the nonenzymic pathway) . M-chlorinecarbonylcyanidephenylhydrazonium, a protonophore uncoupler potassium cyanide, an inhibitor of the respiratory chain, N',N-dicyclohexylcarbodiimide, an inhibitor of ATPase, cause the membrane potential dissipation . The orientation of the transmembrane electric field is as follows: "minus" inside cells and "plus" inside membrane vesicles.

J Antimicrob Chemother, 1987 Nov, 20 Suppl B, 57 - 68
An in-vitro comparison of the intraphagocytic bioactivity of erythromycin and roxithromycin; Anderson R et al.; The binding to human polymorphonuclear leucocytes and the intracellular bioactivity of the macrolide antibiotics erythromycin and roxithromycin on Legionella micdadei, Listeria monocytogenes and Staphylococcus aureus were investigated in vitro by the combination of a fluorochrome microassay and a radioassay . Polymorphs with intact or absent membrane-associated oxidative metabolism were used to investigate the interactions which may occur between the intrinsic oxygen-dependent antimicrobial systems of human polymorphs and the test antibiotics, in the elimination of intracellular microbial pathogens . Elimination of O2-dependent antimicrobial systems with retention of phagocytic activity was achieved by using polymorphs from children with chronic granulomatous disease NaF-pulsed normal polymorphs . Both antimicrobial agents were actively concentrated by polymorphs . Erythromycin was concentrated ten-fold and roxithromycin approximately thirty-fold above extra cellular levels . Both agents possessed intracellular bacteriostatic activity for all three test microbial pathogens . Depletion of polymorph O2-dependent intrinsic antimicrobial systems interfered with the intracellular bioactivity of both antibiotics . This emphasizes the importance of interactions between cell-associated antibiotics and phagocyte antimicrobial systems in the elimination of intracellular microbial pathogens . Like erythromycin, roxithromycin is concentrated by human phagocytes and is bioactive intracellularly.

J Antimicrob Chemother, 1987 Nov, 20(5), 753 - 8
The emergence of resistance to ciprofloxacin during treatment of experimental Staphylococcus aureus endocarditis; Kaatz GW et al.; The efficacy of ciprofloxacin was compared with that of vancomycin in the rabbit model of methicillin-susceptible Staphylococcus aureus endocarditis . Animals were treated with ciprofloxacin, 25 mg/kg iv every 8 h or vancomycin, 17.5 mg/kg iv every 6 h, for 3, 6, or 9 days . Both drugs were found to be equally effective in the therapy of this infection, but the degree of reduction in bacterial counts was less than expected on the basis of previous studies . Additionally, resistance to ciprofloxacin in the test strain of S . aureus was seen to emerge in 12.5% of animals that received the drug . This raises concern about the use of ciprofloxacin as a single agent in the therapy of humans with serious systemic S . aureus infections.

J Antimicrob Chemother, 1987 Nov, 20(5), 713 - 8
Metronidazole and spiramycin in abscesses caused by Bacteroides spp . and Staphylococcus aureus in mice; Brook I; The activity of metronidazole and spiramycin, singly or in combination, was tested in vitro and also in vivo, in the eradication of infection caused by Bacteroides spp . and Staphylococcus aureus, alone or in combination . The MICs of metronidazole for the B . melaninogenicus and B . fragilis strains were significantly reduced by the addition of spiramycin (from 0.25 and 0.5 mg/l, to 0.062 and 0.125 mg/l, respectively) . Antimicrobial synergy between metronidazole and spiramycin was noted against Bacteroides spp . in abscesses caused in mice by subcutaneous injection of Bacteroides spp . alone or in combination with S . aureus . Furthermore, an additional reduction in the number of S . aureus was noted in mixed infections with bacteroides that were treated with metronidazole alone . The antimicrobial synergy in vitro and in vivo between metronidazole and spiramycin may have clinical implications that deserve to be further investigated.

J Antimicrob Chemother, 1987 Nov, 20(5), 705 - 12
Comparative efficacy of clindamycin, erythromycin and spiramycin against Staphylococcus aureus in the rat croton oil pouch model; Davey PG et al.; Spiramycin, a macrolide antibiotic, has inferior in-vitro activity to erythromycin, but superior tissue penetration . Recent publications have suggested that the in-vivo activity of spiramycin should be re-assessed . The efficacy of clindamycin, erythromycin and spiramycin was compared against Staphylococcus aureus infections in the rat croton oil pouch model . The concentration of spiramycin in the pouch fluid was lower than the concentration of clindamycin or erythromycin after single or multiple intraperitoneal injections . In contrast, the concentration of spiramycin in the pouch wall (73.3 +/- 14.5 micrograms/g) was markedly higher than that of erythromycin (less than 7.5 micrograms/g) . Multiple doses of spiramycin had no significant effect upon bacterial growth in the pouch, whereas clindamycin and erythromycin had a significant bactericidal effect . The results suggest that spiramycin is bound to tissues, diffuses poorly into tissue fluid and may therefore be ineffective against infections in large collections of tissue fluid.

J Antimicrob Chemother, 1987 Nov, 20(5), 685 - 95
Cephalothin clearance of Staphylococcus aureus from two experimental infection sites in the presence and absence of local phagocytic cells; Gerding DN et al.; The clearance of Staphylococcus aureus from perforated peritoneal capsules which are accessible to phagocytic cells, and subcutaneous Visking chambers which exclude phagocytes, was studied simultaneously in eight rabbits implanted with both devices . Animals were treated with cephalothin, 100 mg/kg im every 8 h for sixteen doses, beginning 24 h after inoculation of the infection sites with S . aureus (cephalothin MIC 0.125 mg/l, MBC 0.5 mg/l) . At the start of cephalothin, subcutaneous chambers contained a higher concentration of S . aureus (8.4 log10 cfu/ml) than peritoneal capsules (6.8 log10 cfu/ml, P less than 0.001) . There was a significant bactericidal effect in subcutaneous chambers on both the third and sixth day of treatment (P less than 0.002), whereas in peritoneal capsules this did not occur until day six . The total reduction in bacterial count in subcutaneous chambers (7.0 log10 cfu/ml) was significantly greater than in capsules (4.4 log10 cfu/ml, P less than 0.002) . The mean concentration of cephalothin in subcutaneous chambers (10.7 mg/l) was significantly higher than in peritoneal capsules (6.1 mg/l, P less than 0.02), but no difference in in-vitro killing of S . aureus was detected at these concentrations . We conclude that cephalothin clearance of S . aureus from a site accessible to phagocytes was delayed when compared to a phagocyte-inaccessible site.

Appl Environ Microbiol, 1987 Nov, 53(11), 2675 - 6
Detection of low-enterotoxin-producing Staphylococcus aureus strains; Kokan NP et al.; Culture supernatant fluids from 26 (23.6%) monkey feeding test-positive Staphylococcus aureus strains, negative for enterotoxins by gel diffusion, were positive by enzyme-linked immunosorbent assay for one or more of the identified enterotoxins . Staphylococcal enterotoxin D (SED) was produced by 23 (88.5%) strains, SED and SEA were produced in two strains, and SED and SEC were produced in one strain . One strain produced only SEA, and two strains produced only SEC.

Vet Immunol Immunopathol, 1987 Nov, 16(3-4), 185 - 99
Bovine leukocyte phagocytosis and bacteria killing monitored by intracellular acridine orange fluorescence and extracellular fluorescence quenching; Zanetti M et al.; The time course of phagocytosis and intracellular killing of serum-opsonized Escherichia coli K12 and Staphylococcus aureus SG511 by glass-adherent bovine peripheral blood polymorphonuclear leukocytes (PMNLs) and cultured monocytes (macrophages) was monitored by fluorescence microscopy of single cells using the acridine orange (AO)/crystal violet (CV) technique . After interaction of glass-adherent leukocytes (20, 40, 60 min, 37 degrees C) with opsonized bacteria, cells were stained with the fluorescent dye AO . Living bacteria stained green, dead bacteria stained orange . The addition of CV to AO-stained bacteria quenched the fluorescence of extracellular bacteria only . CV does not penetrate living bovine PMNLs which allows the discrimination of ingested (fluorescent) and extracellular (nonfluorescent) bacteria during attachment and phagocytosis of bacteria by adherent PMNLs . We investigated quantitatively phagocytosis and intracellular killing of serum-opsonized bacteria by bovine PMNLs from 22 bulls of 4 different Swiss dairy breeds . Within 60 min maximum uptake (approximately 12 bacteria/PMNL) and killing (approximately 80%) of serum-opsonized Escherichia coli K12 and Staphylococcus aureus SG511 were achieved . The AO/CV technique was also used to quantify the uptake and intracellular killing of serum-opsonized Escherichia coli K12 by cultured monocytes (macrophages) . Within 60 min maximum uptake of bacteria (approximately 16/MO) was achieved; approximately 83% of bacteria were killed.

Clin Exp Immunol, 1987 Nov, 70(2), 463 - 70
In vitro analysis of B lymphocyte function in uraemia; Degiannis D et al.; We have investigated the immune responses in vitro of uraemic patients undergoing regular haemodialysis or continuous ambulatory peritoneal dialysis . Twenty-five healthy subjects were also studied as controls . In uraemic patients, the number of T and B lymphocytes were within the normal range, but proliferative responses to phytohaemagglutinin (PHA) were impaired . Spontaneous immunoglobulin plaque forming cell (PFC) responses by peripheral blood mononuclear cells (PBMC) from uraemic patients were significantly lower than those of healthy subjects . The PFC response of uraemic PBMC to the T cell independent polyclonal B cell activator (PBA) Epstein-Barr virus (EBV) was comparable to the response of the healthy subjects, indicating that uraemic B cells are still capable of synthesizing immunoglobulin . Pokeweed mitogen (PWM) induced PFC responses of uraemic PBMC were also normal, whereas the response to another T cell dependent B cell activator, Staphylococcus aureus Cowan I (SAC), was very low . Addition of indomethacin to PWM- and SAC-activated cultures of uraemic PBMC enhanced the PFC response to SAC, but had little effect on the PWM response . As full differentiation of B cells in response to SAC depends on helper T cells, we conclude that a defect in T lymphocyte function accounts for the reduced spontaneous and SAC induced production of immunoglobulin by uraemic PBMC . This defect may be mediated by an indomethacin-sensitive mechanism.

Otolaryngol Clin North Am, 1987 Nov, 20(4), 853 - 76
Postoperative sequelae and complications of rhinoplasty; Holt GR et al.; This article has overviewed complications of rhinoplasty . Generally, these complications fall into two categories: aesthetic (that is, cosmetic sequelae that may require a revision rhinoplasty) and nonaesthetic . Of the nonaesthetic complications, infection has the widest span of severity . A localized Staphylococcus aureus abscess or Pseudomonas infection of the nose may occur postoperatively . Owing to the proximity of the nose to the cranium, a cavernous sinus thrombosis or basilar meningitis may result . Postoperative toxic-shock syndrome is a rare occurrence that surgeons should be aware of; most cases have occurred with the presence of nasal packing, but a case using only plastic nasal splints has been reported also . Bacteremia seems to be uncommon during rhinoplasty . Infection after rhinoplasty is generally much less frequent than one would expect from an operation in an unsterile field . Antibiotics are frequently utilized electively . Postoperative nasal-periorbital edema and ecchymosis are regarded as unavoidable but may be lessened significantly by postoperative head elevation and cold packs . The possibility of postoperative bleeding must be evaluated by the surgeon preoperatively . This sequela usually occurs either within 72 hours postoperatively or at around 10 days postoperatively . Many different causes exist for chronic postoperative nasal obstruction, from poorly supported nasal valves closing upon inspiration to an enhanced allergic rhinitis leading to chronic nasal mucosal edema . The latter may be treated by injection of steroid into the turbinates . Among aesthetic complications, supratip prominence, saddle deformity, and persistent hump are among the more commonly reported . Supratip prominence--"polly-beak"--can be caused by inadequate reduction of tip cartilaginous or soft-tissue elements, especially in relation to the reduction of the dorsum . An over-reduced dorsum will leave an otherwise normal nasal tip with a relative prominence . An accumulation of blood or a mucous cyst occurring under the skin of the tip will produce a prominence . Poor tip projection, tip ptosis, and alar collapse are the result of overreduction of tip elements . A dislocated alar cartilage can appear as an asymmetric nasal bossa . Saddle-nose deformity occurs after overaggressive bony and/or cartilaginous hump removal . Infractured nasal bones that subsequently drop into the piriform aperture can create a bony saddle . Persistent hump is due to inadequate reduction of a bony or cartilaginous hump . If the septal cartilage reduction is disproportionate to the bony septum reduction, the appearance of either a hump or a saddle is possible.(ABSTRACT TRUNCATED AT 400 WORDS)

J Clin Microbiol, 1987 Nov, 25(11), 2163 - 7
Legionella micdadei and Legionella dumoffii monoclonal antibodies for laboratory diagnosis of Legionella infections; Cercenado E et al.; Two different monoclonal antibodies directed against Legionella micdadei and L . dumoffii (Genetic Systems Corp., Seattle, Wash.) were evaluated for their specificity and ability to detect L . micdadei and L . dumoffii in human and animal clinical samples and bacterial isolates in an indirect immunofluorescence assay . All three frozen sputum samples and all three Formalin-fixed sputum and liver samples from patients with culture-documented L . micdadei pneumonia were positive when tested with the L . micdadei monoclonal antibody . A Formalin-preserved lung sample from a patient with culture-documented L . dumoffii pneumonia was positive with its homologous monoclonal antibody . No cross-staining reactions were found with either monoclonal antibody on any of 25 human sputum samples tested from patients without Legionella infections . A total of 66 Legionella strains and 56 non-Legionella strains including 22 Pseudomonas strains and 34 other bacterial strains were studied . No cross-staining reactions were found except in Staphylococcus aureus Cowan 1 ATCC 12598 . The lower limit of detection in seeded sputum samples was about 7 X 10(4) cells per ml for both monoclonal antibodies . Lung and tracheal lavage specimens from L . micdadei- or L . dumoffii-infected guinea pigs showed specific staining only with their respective monoclonal antibodies . The monoclonal antibodies stained homologous bacteria slightly less intensely than did the polyclonal antisera, but the signal-to-noise ratio was considerably higher for the monoclonal antibodies . No differences in sensitivity of staining of clinical specimens or bacterial isolates were noted between the monoclonal antibodies and the polyclonal reagents for L . micdadei and L . dumoffii (Centers for Disease Control, Atlanta, Ga., and BioDx, Denville, N.J . These monoclonal antibodies ae sensitive and specific, making them good candidates for laboratory diagnostic purposes.

Neurology, 1987 Nov, 37(11), 1747 - 53
Cervical epidural abscess; Lasker BR et al.; We present 3 new cases of cervical epidural abscess (CEA), a rare condition, along with a review of 12 other case reports . The average patient age was 45 years; just over half were male . The abscesses usually involved the mid to lower cervical region and extended an average of three to four segments . Neck stiffness was present in all patients; root pain and paresthesias were present less often . Weakness of one to four extremities developed in all but one patient . Sensory levels were frequently present, sometimes below the site of the lesion . All but two patients were febrile . All but two had elevated CSF protein, and all but two had a pleocytosis; myelography always revealed a complete or partial block . Staphylococcus aureus was the causative organism in 8 of 11 patients . CEA should be considered in a patient with neck stiffness, paresthesias, and/or radicular pain so that CT or myelography followed by surgical decompression and/or antimicrobial drugs can be initiated before prolonged weakness develops . One of our patients developed a syrinx causing a new neurologic deficit 3 years after treatment . Delayed syringomyelia, a rare complication of extramedullary lesions, lends support to vascular occlusion as the major mechanism of damage in epidural abscess.

Cardiovasc Res, 1987 Nov, 21(11), 813 - 20
Bacterial tissue tropism: an in vitro model for infective endocarditis; Becker RC et al.; Since infective endocarditis may affect individuals without pre-existing valvar heart disease, and Staphylococcus aureus is the organism most commonly involved, the binding characteristics of S aureus to several components of normal vascular endothelium and subendothelium were studied . S aureus adhered specifically to endothelial monolayers (6.08(1.10)%; p less than 0.005), fibronectin (5.43(0.81)%; p less than 0.001), fibrinogen (7.13(1.43)%; p less than 0.001), and acid soluble calf skin collagen (2.38(0.90)%; p less than 0.001) . S aureus also adhered specifically to Von Willebrand factor (1.62(0.28)%, p less than 0.001) . Protein A containing (Cowan I) and deficient (Wood) strains of S aureus adhered similarly to all surfaces and substrates (NS) . Escherichia coli adhered poorly . Immunofluorescence microscopy of preconfluent endothelial cells identified an extensive pericellular fibronectin network at regions of cell to cell contact . Light microscopy showed S aureus binding solely within these regions . Therefore, the ability of S aureus to infect valvar endothelium may be dependent on the presence of a fibronectin receptor . The existence of specific receptor for S aureus on the endothelial cell surface itself remains undetermined.

Somat Cell Mol Genet, 1987 Nov, 13(6), 661 - 9
Overlapping transcription units in the transient receptor potential locus of Drosophila melanogaster; Wong F et al.; We report the identification in Drosophila melanogaster of two mRNA transcripts that are derived from the transient receptor potential locus by transcription in opposite directions . The two transcripts overlap; one transcript has, as part of its 5'-untranslated sequence, the reverse complement of 442 bp of the 3' terminus of the transcript derived from the opposite DNA strand . Conceptual translation of the corresponding cDNA sequences predicted for one of the transcripts a polypeptide whose C terminus shares sequence and structural similarity with the cell-wall-binding domain of protein A from Staphylococcus aureus; for the transcript derived from the opposite DNA strand, a polypeptide of 264 amino acids was predicted, which showed no significant sequence homology with any known protein . The two transcripts have different tissue specificities: one is expressed predominantly in the eye and the other is in the body . These findings may have implications in the relationship between the organization of overlapping genes on opposite DNA strands and regulation of gene expression by antisense RNA.

J Immunol, 1987 Nov 1, 139(9), 2970 - 6
Enhancement of human B cell proliferation and differentiation by tumor necrosis factor-alpha and interleukin 1; Jelinek DF et al.; The role of tumor necrosis factor-alpha (TNF-alpha) in human B cell responses was examined and compared with that of interleukin (IL) 1 by assessing the ability of each cytokine to support proliferation and differentiation . Recombinant TNF-alpha (rTNF-alpha) and recombinant IL-1 (rIL-1) each enhanced the generation of immunoglobulin-secreting cells (ISC) in cultures of pokeweed mitogen-stimulated B cells incubated with T cells . To examine the direct effect of rTNF-alpha and rIL-1 on the responding B cell, highly purified peripheral blood B cells were stimulated with Cowan I Staphylococcus aureus (SA) . In the absence of T cell factors, proliferation was minimal and there was no generation of ISC . Recombinant IL-2 (rIL-2) supported both responses . Although rTNF-alpha alone did not support SA-stimulated generation of ISC, it did increase SA-stimulated B cell DNA synthesis by two- to eightfold . In addition, rTNF-alpha augmented B cell proliferation in rIL-2 supported SA-stimulated cultures . Moreover, rTNF-alpha enhanced the generation of ISC stimulated by rIL-2 alone or rIL-2 and SA . rIL-1 also augmented DNA synthesis and generation of ISC by B cells stimulated with SA and rIL-2 . However, rTNF-alpha enhanced proliferation and ISC generation in SA + rIL-2-stimulated cultures even when they were supplemented with saturating concentrations of rIL-1 . Utilizing a two-stage culture system, it was found that the major effect of rTNF-alpha was to enhance responsiveness of SA-activated B cells to rIL-2, whereas it exerted only a minimal effect during initial stimulation . These results indicate that TNF-alpha as well as IL-1 augment B cell responsiveness . Moreover, the ability of rTNF-alpha to enhance B cell responsiveness was not an indirect effect resulting from the induction of Il-1 production by contaminating monocytes, but rather resulted from the delivery of a signal by rTNF-alpha directly to the responding B cell that promoted both proliferation and differentiation after initial activation . The data therefore indicate that human B cell responsiveness can be independently regulated by the action of two separate monocyte-derived cytokines.

Blood, 1987 Nov, 70(5), 1679 - 82
Localization of a factor VIII binding domain on a 34 kilodalton fragment of the N-terminal portion of von Willebrand factor; Takahashi Y et al.; Factor VIII (F.VIII) was tested for its ability to bind in solid phase system to von Willebrand Factor (vWF) or fragments obtained with Staphylococcus aureus V-8 protease, ie, SpIII (N-terminal), SpI (central), and SpII (C-terminal) . Bound F.VIII was estimated in situ by clotting and chromogenic assays . F.VIII bound in a dose-dependent manner to immobilized vWF and SpIII but not to SpII or SpI . Binding was inhibited by 0.25 mol/L CaCl2 as well as by an excess of vWF or SpIII . Accordingly, immobilized F.VIII specifically bound 125I-vWF and SpIII but not SpII or SpI . Twelve monoclonal antibodies (MoAbs) directed towards SpIII, specifically blocking binding of F.VIII to vWF or SpIII, were used for the mapping of plasmic or tryptic fragments of vWF or SpIII . We thus established that a F.VIII binding domain of vWF is located on a 34 kilodalton (kd) fragment of the N-terminal portion of vWF, between residues 1 and 910, and that it is distinct from the GPIb and collagen binding domains.

J Hosp Infect, 1987 Nov, 10(3), 255 - 9
Methicillin resistant Staphylococcus aureus; the role of antisepsis in the control of an outbreak; Tuffnell DJ et al.; Between February 1983 and September 1985, an outbreak of methicillin-resistant Staphylococcus aureus involving 151 patients and staff occurred in a district general hospital . At its peak, 43 cases occurred in 3 months . Sixty-two patients suffered morbidity and two died . Conventional isolation techniques and once-daily whole body washing of affected patients with triclosan successfully controlled the outbreak.

Blood, 1987 Nov, 70(5), 1577 - 83
Localization of a collagen-interactive domain of human von Willebrand factor between amino acid residues Gly 911 and Glu 1,365; Kalafatis M et al.; A collagen-binding domain of von Willebrand factor (vWF) has been identified in the central part of the molecule by comparing the binding properties of vWF and Staphylococcus aureus V-8 protease-generated vWF fragments with collagen . The binding of purified human vWF to human type III collagen was found to be specific . At saturation, 38 to 50.2 micrograms of vWF bound per milligram of collagen . Scatchard plots derived from binding isotherms demonstrated the presence of at least two classes of binding sites . Purified vWF was digested with S aureus V-8 protease into two complementary fragments (SpIII and SpII) . SpII, the C-terminal end of vWF (amino acid residues 1,366 to 2,050), was totally devoid of affinity for collagen . Contrarily, purified SpIII, the N-terminal part of vWF (residues 1 to 1,365), totally displaced vWF binding and specifically bound to collagen . At saturation, 25 to 45 micrograms of SpIII bound per milligram of collagen . Scatchard plots demonstrated the presence of a single class of binding sites . SpIII was further digested with the same enzyme to generate SpI, a 52-kilodalton fragment from the C-terminal part of SpIII (residues 911 to 1,365) . Spl induced a dose-dependent inhibition of both vWF and SpIII binding to collagen . A series of six monoclonal antibodies against SpIII that completely abolished vWF and SpIII interaction with collagen also bound to SpI . In conclusion, SpI extending between amino acid residues 911 and 1,365 of vWF contains a specific site that interacts with human type III collagen.

J Gen Microbiol, 1987 Nov, 133 ( Pt 11), 3039 - 52
The aacA-aphD gentamicin and kanamycin resistance determinant of Tn4001 from Staphylococcus aureus: expression and nucleotide sequence analysis; Rouch DA et al.; The aacA-aphD aminoglycoside resistance determinant of the Staphylococcus aureus transposon Tn4001, which specifies resistance to gentamicin, tobramycin and kanamycin, has been cloned and shown to express these resistances in Escherichia coli . The determinant encoded a single protein with an apparent size of 59 kDa which specified both aminoglycoside acetyltransferase {AAC(6')} and aminoglycoside phosphotransferase {APH(2")} activities . Nucleotide sequence analysis of the determinant showed it to be capable of encoding a 479-amino-acid protein of 56.9 kDa . analysis of Tn1725 insertion mutants of the determinant indicated that resistance to tobramycin and kanamycin is due to the AAC activity specified by, approximately, the first 170 amino acids of the predicted protein sequence and is consistent with the gentamicin resistance, specified by the APH activity, being encoded within the C-terminal region of the protein . Comparison of the C-terminal end of the predicted amino acid sequence with the reported sequences of 13 APHs and a viomycin phosphotransferase revealed a region which is highly conserved among these phosphotransferases.

J Gen Microbiol, 1987 Nov, 133 ( Pt 11), 3031 - 8
Detection and characterization of IS256, an insertion sequence in Staphylococcus aureus; Lyon BR et al.; Resistance to the aminoglycosides gentamicin (Gmr), tobramycin (Tmr) and kanamycin (Kmr) in strains of Staphylococcus aureus isolated in Australia is mediated by the transposon Tn4001 . The 1.35 kb inverted repeat of this transposon exhibits many of the characteristics of an insertion sequence and has consequently been designated IS256 . Tandem duplication of IS256 contiguous with Tn4001 results in an increase in the level of GmrTmrKmr, thereby implying that the element possesses strong promoter sequences . Both contiguous and independent insertions of IS256 into the staphylococcal chromosome have been observed, the latter suggesting that the element may play a role in molecular rearrangements of the genome.

EMBO J, 1987 Nov, 6(11), 3253 - 60
A recombinant apoA-1--protein A hybrid reproduces the binding parameters of HDL to its receptor; Monaco L et al.; We have constructed a plasmid, pLM8, containing the coding sequence of the mature human apoA-1 fused to the coding sequence of the IgG-binding domains of protein A (PA) from Staphylococcus aureus . The hybrid gene is transcribed in Escherichia coli under the control of a heat-sensitive repressor, leading to the synthesis of large amounts of hybrid protein (apoA-1--PA) . The hybrid protein was purified by denaturation with urea and alkali, renaturation and affinity chromatography on an IgG Sepharose column . ApoA-1--PA is soluble and has an Mr of 316 kd, as determined by gel filtration . This is five times the monomer size of 62 kd, predicted from the sequence and found by SDS-PAGE analysis . Cell surface binding activity of the hybrid protein was tested using two different cell types (J774 macrophages and Fao hepatocytes) and compared to human high density lipoprotein (HDL) . High-affinity binding was found for both ligands in both cell lines (Kd = 3.4 X 10(-8)M in Fao cells, 4.9 X 10(-8) M in J774 cells for apoA-1--PA and 3.0 X 10(-8) M in Fao cells, 2.8 X 10(-8) M in J774 cells for HDL), with approximately 2 X 10(5) high-affinity binding sites per cell . ApoA-1--PA and HDL effectively competed with each other for binding to the cell surface . Additionally, they both bound to a 110-kd polypeptide on a ligand blot, identifying an HDL receptor . The binding parameters of HDL were very similar to those of apoA-1--PA.(ABSTRACT TRUNCATED AT 250 WORDS)

Proc Natl Acad Sci U S A, 1987 Nov, 84(22), 7967 - 71
A series of six ligands for the human formyl peptide receptor: tetrapeptides with high chemotactic potency and efficacy; Rot A et al.; We recently isolated, from culture fluids of Staphylococcus aureus, a chemotactic peptide that comprised equimolar quantities of methionine, leucine, phenylalanine, and isoleucine . It interacted with the formylmethionyl peptide receptor of human leukocytes and had considerably higher potency and efficacy than the widely studied tripeptide agonist fMet-Leu-Phe . On the assumption that the attractant was a formylmethionyl tetrapeptide, we synthesized the six possible sequences and tested the products for chemotactic potency and efficacy, as well as their capacity to inhibit binding of fluorescein isothiocyanate-labeled fMet-Leu-Phe-Lys to human monocytes . The concentrations required for inhibition of fluorescein-labeled fMet-Leu-Phe-Lys binding by the six peptides covered three orders of magnitude . Chemotactic potency (concentration that caused 50% of the maximum chemotactic response) ranged from 3.1 X 10(-11) M to 6.4 X 10(-10) M; efficacy (percentage of monocytes migrating at optimal attractant concentration) ranged from 41% to 66% . When the six synthetic tetrapeptides were ranked for chemotactic efficacy, they paired according to the position of phenylalanine . The average percentage migration was 66% for the two peptides with phenylalanine in position 3, 51% for phenylalanine in position 4, and 41% for phenylalanine in position 2 . Since the published value for the percentage of human monocytes with detectable formyl peptide receptors is 60%, it is apparent that the two tetrapeptides with phenylalanine in position 3 (fMet-Ile-Phe-Leu and fMet-Leu-Phe-Ile) are full chemotactic agonists, which are capable of inducing migration of all the receptor-bearing cells . This is in contrast to the tripeptide fMet-Leu-Phe, which induces migration of only 50% of monocytes with receptors (efficacy of 33%) . Since the chemotactic efficacy of the six tetrapeptides covers a wide range, the series may be useful to investigate signals that lead to directed movement after occupancy of receptors by chemoattractants.

Biochem Pharmacol, 1987 Nov 1, 36(21), 3763 - 70
Polyclonal antibodies to agonist benzodiazepines; Fry JP et al.; Benzodiazepine-binding, immunoglobulin G class antibodies have been raised in three rabbits immunised with a conjugate of kenazepine coupled to keyhole limpet haemocyanin . The antibodies were assayed by {3H}flunitrazepam binding, followed by adsorption onto Staphylococcus aureus cells . Measurement of the rates of association and dissociation of {3H}flunitrazepam binding, together with saturation analysis of equilibrium binding, revealed varying degrees of heterogeneity in the affinity constants of the three rabbit antisera (equilibrium KD values 0.18 to 4.13 nM at 20-22 degrees) . Specificity of the antibodies was investigated by testing a wide variety of compounds (at concentrations of up to 10-100 microM) for their ability to inhibit {3H}flunitrazepam binding . Only benzodiazepines known to act as agonists at their receptor sites in the central nervous system (CNS) caused an inhibition of binding . The rank orders of the IC50 values of these drugs for inhibition of {3H}flunitrazepam binding to IgG from two out of the three rabbits correlated significantly with that previously published for displacement of CNS receptor binding . The agonist beta-carboline derivative ZK 93423, the anxiolytic cyclopyrrolones suriclone and zopiclone and the purines inosine and hypoxanthine all failed to inhibit antibody binding, supporting previous suggestions that these drugs may bind at non-benzodiazepine recognition sites on the CNS receptor . The antibodies described are expected to provide useful reagents for raising anti-idiotypic antibodies directed against the CNS receptor and for the identification and purification of possible endogenous benzodiazepine receptor agonists in the CNS.

Ann Surg, 1987 Nov, 206(5), 578 - 82
Impaired nonspecific cellular immunity in experimental cholestasis; Roughneen PT et al.; The abilities of polymorphonuclear leukocytes (PMN) and pulmonary alveolar macrophages (PAM), to demonstrate chemotaxis, phagocytosis, and superoxide release after bile duct ligation in the rat were investigated to determine the effect of cholestasis on nonspecific cellular immune mechanisms . Chemotactic response to C5a and FMLP, phagocytosis of 14C labeled Staphylococcus aureus, and zymosan-induced superoxide release were evaluated 21 days after bile duct ligation (BDL), sham operation, or in normal controls . Serum total bilirubin level was elevated after BDL (p less than 0.01) . Chemotactic ability was similar to each group . PMN phagocytic uptake of 14C labeled Staphylococcus aureus was depressed in BDL (p less than 0.05) . BDL rats exhibited impaired PAM phagocytic indices and improved PMN superoxide release (p less than 0.03) . PAM superoxide release was similar in each study group . Alterations in phagocytic function with cholestasis are important deficits in nonspecific cellular immunity that may contribute to the high incidence of infective complications associated with obstructive jaundice.

J Infect Dis, 1987 Nov, 156(5), 741 - 50
Virulence studies, in mice, of transposon-induced mutants of Staphylococcus aureus differing in capsule size; Lee JC et al.; We used three related strains of Staphylococcus aureus to determine whether capsule size influenced bacterial virulence . Strain SA1 mucoid elaborated a large capsule demonstrable by transmission electron microscopy (TEM) . Nonmucoid isolates were derived from strain SA1 mucoid by Tn551 insertional mutagenesis . By TEM, strain JL24 produced a "microcapsule," whereas strain JL25 was unencapsulated . Strain SA1 mucoid had a 50% lethal dose for mice greater than 3,000-fold lower than that of strains JL24 and JL25 . Quantitative cultures of blood and kidney from animals challenged intravenously revealed that strain SA1 mucoid was cleared less readily from the bloodstream and kidneys than the nonmucoid mutants . In an in vitro assay, only strain SA1 mucoid demonstrated antibody-dependent, complement-mediated opsonophagocytosis by human leukocytes . Strains JL24 and JL25 were opsonized for phagocytosis by complement alone . Thus a highly encapsulated strain of S . aureus was more virulent in mice than two related nonmucoid strains . The microencapsulated mutant was not more virulent than the unencapsulated mutant.

J Immunol, 1987 Nov 1, 139(9), 2942 - 6
Epitopes on proliferating cell nuclear antigen recognized by human lupus autoantibody and murine monoclonal antibody; Ogata K et al.; The immune epitopes of proliferating cell nuclear antigen (PCNA), also called cyclin, were analyzed by determining the reactivity between PCNA peptide fragments and anti-PCNA antibodies from lupus patients, murine monoclonal antibody (19A2), and rabbit anti-NH2-terminal peptide antibody . Limited digestion of PCNA/cyclin with Staphylococcus aureus V8 protease resulted in several peptide fragments . Five fragments of 30, 20, 15, 14, and 13 kDa were reactive with rabbit anti-NH2-terminal peptide antibody denoting that they contained the NH2-terminal peptide . The 30- and 20-kDa fragments reacted with 19A2 but the others did not . Lupus sera reacted with 17- and 15-kDa peptide fragments allowing their classification into three groups . Two of eight sera (type A) reacted only with the 17-kDa fragment . Two others (type B) reacted with both the 17- and 15-kDa fragments and the remaining four sera (type C) reacted only with the 15-kDa fragment . The sera reacting with the 15-kDa fragment also reacted with the 20-kDa fragment, but the sera reactive only with the 17-kDa fragment did not, indicating that the 17-kDa fragment was not a degradation product of 20-kDa fragments . The 19A2 epitope resided in the region between 15 and 20 kDa from the NH2 terminus, whereas there was at least one distinct epitope on each 15- and 17-kDa peptide, which were recognized by lupus autoantibodies.

Biochem Biophys Res Commun, 1987 Oct 29, 148(2), 658 - 63
Characterization and interactions of a fragment of the core protein of the small proteoglycan (PGII) from bovine tendon; Vogel KG et al.; Sequence analysis showed that Staphylococcus aureus V8 protease cleaved the core protein of the small dermatan sulfate proteoglycan of bovine tendon (PGII) on the carboxy side of a glutamic acid residue located 17 amino acids from the N-terminus of the intact molecule . The remaining 40 kDa core protein fragment inhibited collagen fibrillogenesis in an in vitro assay . V8 protease readily generated this fragment in tendon tissue, but it was not released from the tissue during treatment . These results indicate that neither the 17-amino acid N-terminal peptide nor the glycosaminoglycan chain attached to this peptide is required for maintaining the interaction of this proteoglycan with a collagen matrix.

J Biol Chem, 1987 Oct 15, 262(29), 14260 - 4
Isolation and characterization of an altered cytochrome P-450 from a yeast mutant defective in lanosterol 14 alpha-demethylation; Aoyama Y et al.; A cytochrome P-450 (P-450SG1) was purified from a lanosterol 14 alpha-demethylase (P-450(14DM)) defective mutant of Saccharomyces cerevisiae, strain SG1, by a method similar to that used in the purification of the wild type enzyme (Yoshida, Y., and Aoyama, Y . (1984) J . Biol . Chem . 259, 1655-1660) . P-450SG1 had the same apparent Mr as and was immunochemically identical to P-450(14DM) . Peptide maps of P-450SG1 made by limited proteolysis with Staphylococcus aureus V8 proteinase, chymotrypsin, or papain followed by gel electrophoresis were identical to corresponding peptide maps of P-450(14DM) . However, P-450SG1 showed no lanosterol 14 alpha-demethylase activity and its mode of interaction with diniconazole {(E)-1-(2,4-dichlorophenyl)-4,4-dimethyl-2-(1,2,4-triazol-1-y1)-1- penten-3- o1}, a specific inhibitor of P-450(14DM), was fundamentally different from that of P-450(14DM) . The absorption spectrum of ferric P-450SG1 was unusual for a native low-spin cytochrome P-450 and was superimposable on that of 1-methylimidazole complex of P-450(14DM), indicating that P-450SG1 has a histidine 6th ligand trans to the thiolate 5th ligand, while the 6th ligand of other ferric low-spin cytochrome P-450s is a water molecule or a hydroxyl group of an oxyamino acid . It is concluded that P-450SG1 is an altered P-450(14DM) . Difference in the primary structure between P-450SG1 and P-450(14DM) may be slight and was not detected by peptide mapping . However, the alteration caused significant change in the substrate site and heme environments of the cytochrome . P-450SG1 is the first example of a cytochrome P-450 having a histidine axial ligand trans to thiolate and of a genetically altered cytochrome P-450 isolated in a homogeneous state.

Biochim Biophys Acta, 1987 Oct 15, 915(3), 331 - 41
Frequency-domain fluorescence studies of an extracellular metalloproteinase of Staphylococcus aureus; Wasylewski Z et al.; A frequency-domain fluorescence study of calcium-binding metalloproteinase from Staphylococcus aureus has shown that this two-tryptophan-containing protein exhibits a double-exponential fluorescence decay . At 10 degrees C in 0.05 M Tris-HCl buffer (pH 9.0) containing 10 mM CaCl2, fluorescence lifetimes of 1.2 and 5.1 ns are observed . Steady-state and frequency-domain solute-quenching studies are consistent with the assignment of the two lifetimes to the two tryptophan residues . The tryptophan residue characterized by a shorter lifetime has a maximum of fluorescence emission at about 317 nm and the second one exhibits a maximum of its emission at 350 nm . These two residues contribute almost equally to the protein's fluorescence . These results, as well as fluorescence-quenching studies using KI and acrylamide as a quencher, indicate that in calcium-loaded metalloproteinase, the tryptophan residue characterized by the shorter lifetime is extensively buried within the protein . The second residue is exposed on the surface of the protein . The tryptophan residues of metalloproteinase have acrylamide dynamic-quenching rate constants, kq values, of 2.3 and 0.26 X 10(9) M-1 X s-1 for the exposed and buried residue, respectively . A study of the temperature dependence of the fluorescence lifetime for the two tryptophan components gives activation energies, Ea values, for thermal quenching of 1.8 and 2.2 kcal/mol for the buried and the exposed residue, respectively . Dissociation of Ca2+ from the protein causes a change in the protein's structure, as can be judged from dramatic changes which occur in the fluorescence properties of the buried tryptophan residue . These changes include an approx . 13 nm red-shift in the maximum of the fluorescence emission and an increase in the acrylamide-quenching rate constant, and they indicate that the removal of Ca2+ results in an increase in the exposure and the polarity of the microenvironment of this 'blue' residue.

J Immunol, 1987 Oct 15, 139(8), 2749 - 54
Inhibition of monocyte oxidative responses by Bordetella pertussis adenylate cyclase toxin; Pearson RD et al.; Bordetella pertussis and the other Bordetella species produce a novel adenylate cyclase toxin which enters target cells to catalyze the production of supraphysiologic levels of intracellular cyclic adenosine monophosphate (cAMP) . In these studies, dialyzed extracts from B . pertussis containing the adenylate cyclase toxin, a partially purified preparation of adenylate cyclase toxin, and extracts from transposon Tn5 mutants of B . pertussis lacking the adenylate cyclase toxin, were used to assess the effects of adenylate cyclase toxin on human peripheral blood monocyte activities . Luminol-enhanced chemiluminescence of monocytes stimulated with opsonized zymosan was inhibited greater than 96% by exposure to adenylate cyclase toxin-containing extract, but not by extracts from adenylate cyclase toxin-deficient mutants . The chemiluminescence responses to particulate (opsonized zymosan, Leishmania donovani, and Staphylococcus aureus) and soluble (phorbol myristate acetate) stimuli were inhibited equivalently . The superoxide anion generation elicited by opsonized zymosan was inhibited 92% whereas that produced by phorbol myristate acetate was inhibited only 32% by B . pertussis extract . Inhibition of oxidative activity was associated with a greater than 500-fold increase in monocyte cAMP levels, but treated monocytes remained viable as assessed by their ability to exclude trypan blue and continued to ingest particulate stimuli . The major role of the adenylate cyclase toxin in the inhibition of monocyte oxidative responses was demonstrated by: 1) little or no inhibition by extracts from B . pertussis mutants lacking adenylate cyclase toxin; 2) high level inhibition with extract from B . parapertussis, a related species lacking pertussis toxin; and 3) a reciprocal relationship between monocyte cAMP levels and inhibition of opsonized zymosan-induced chemiluminescence using both crude extract and partially purified adenylate cyclase toxin . Pertussis toxin, which has been shown to inhibit phagocyte responses to some stimuli by a cAMP-independent mechanism, had only a small (less than 20%) inhibitory effect when added at concentrations up to 100-fold in excess of those present in B . pertussis extract . These data provide strong support for the hypothesis that B . pertussis adenylate cyclase toxin can increase cAMP levels in monocytes without compromising target cell viability or impairing ingestion of particles and that the resultant accumulated cAMP is responsible for the inhibition of oxidative responses to a variety of stimuli.

Biochemistry, 1987 Oct 6, 26(20), 6483 - 8
Primary structure of Paim I, an alpha-amylase inhibitor from Streptomyces corchorushii, determined by the combination of Edman degradation and fast atom bombardment mass spectrometry; Hirayama K et al.; Paim I, a protein alpha-amylase inhibitor, inhibits animal alpha-amylases from pig, dog, cow, horse, etc . but has no activity against human salivary and pancreatic amylases . The primary structure of Paim I has been determined by Edman degradation and fast atom bombardment mass spectrometry (FABMS) . This protein is a single-chain polypeptide of 73 amino acid residues with a calculated molecular weight from the sequence data of 7415.3 (monoisotopic molecular weight) and 7420.2 (average molecular weight) . The sequencing strategy chosen for Paim I consists of four steps . First, the accurate molecular weights of the intact and tetra-S-carboxymethylated Paim I are determined by fast atom bombardment mass spectrometry . Second, the primary fragments generated by Staphylococcus aureus V8 protease are isolated by reversed-phase high-performance liquid chromatography . The molecular weights of these subpeptides are determined by FABMS . The peptides that must be sequenced are selected by the molecular weights of these subpeptides and the tetra-S-carboxymethylated Paim I . Third, these subpeptides and the whole protein are sequenced by automated Edman degradation . Finally, the primary structure of tetra-S-carboxymethylated Paim I is confirmed by the combination of tryptic, chymotryptic, and S . aureus V8 protease digestion and FABMS . The sequence of Paim I is compared with those of Haim II, Hoe-467A, Z-2685, and AI-3688 because they have different alpha-amylase inhibition spectra against mammalian alpha-amylases but belong to a family of related proteins.

J Biol Chem, 1987 Oct 5, 262(28), 13835 - 41
Isolation and characterization of two domains of human von Willebrand factor that interact with fibrillar collagen types I and III; Pareti FI et al.; We have identified four discrete proteolytic fragments of von Willebrand factor (vWF) that define two collagen-binding domains . Two of the fragments tested, T 96 kDa and T 55 kDa, were generated by digestion with trypsin, and two, Fragments I and III, with Staphylococcus aureus V8 protease . The larger Fragment III, a disulfide-linked homodimer, extends between residues 1 and 1365 of the 2050-residue vWF subunit and comprises the sequence of all the others . T 96 kDa, also a disulfide-linked homodimer, extends between residues 449 and 728 . T 55 kDa and Fragment I, both single-chain polypeptides, have a partial sequence overlap corresponding to residues 911-1114, and together extend from residue 730 to 1365 . The ability of the fragments to interfere with the vWF-collagen interaction was evaluated by measuring inhibition of 125I-labeled vWF binding to fibrillar bovine collagen types I and III . All the four fragments tested inhibited binding . Native conformation was essential for expression of this function; denaturation with guanidine hydrochloride and reduction of disulfide bonds resulted in marked reduction or complete loss, respectively, of the inhibitory activity at all the concentrations tested . Two monoclonal antibodies were prepared, one directed against T 96 kDa and the other against Fragment I . Both antibodies partially inhibited vWF binding to collagen, and their inhibitory effect was enhanced when they were used together . 125I-Labeled Fragment I bound to collagen in a saturable manner, and the binding was completely blocked by both T 96 kDa and T 55 kDa . Thus, we have identified at least two distinct functional domains of vWF that concurrently mediate the vWF-collagen interaction . The two domains appear to share a common recognition site on collagen.

S Afr Med J, 1987 Oct 3, 72(7), 476 - 7
Microbiology of secondary osteomyelitis . Value of bone biopsy; Jacobson IV et al.; Secondary osteomyelitis occurs as a direct infection of bone from a source outside the body or as progressive and continuous spread of infection from a contiguous focus . Twenty patients with secondary osteomyelitis were studied . Pathogens were identified by culture of biopsy material from infected bone . Staphylococcus aureus accounted for most cases in the upper limbs, while infection in the lower limbs was predominantly associated with Gram-negative bacilli . The essential value of bone biopsy in treatment is emphasised.

Microbiologica, 1987 Oct, 10(4), 345 - 51
Intensive care units as a source of methicillin-resistant Staphylococcus aureus; Corticelli AS et al.; Over a 12 month period, 209 isolates of methicillin resistant Staphylococcus aureus (MRSA) were obtained in 39 patients admitted to an ICU . In 23 patients MRSA was the major pathogen, producing either pneumonia, bacteremia or wound infection . In eight patients death was directly related to the MRSA infection . This study suggests an increasing occurrence of MRSA infections in ICU and the need to adopt control measures.

J Antimicrob Chemother, 1987 Oct, 20(4), 599 - 608
Determination of methicillin-resistance in Staphylococcus aureus by agar dilution and disc diffusion methods; French GL et al.; Four-hundred and seventy-six isolates of Staphylococcus aureus from patients in Hong Kong were tested for methicillin-resistance by agar dilution and disc diffusion methods, using heavy inocula . With Mueller-Hinton agar incubated at 30 degrees C for 24 h, 216 (MRSA) isolates were resistant to 8 mg/l of methicillin and grew up to the edge of a 10 micrograms methicillin disc, and 260 strains were susceptible to greater than or equal to 4 mg/l methicillin and produced inhibition zones of at least 20 mm diameter . Incubation for 48 h, the addition of sodium chloride, or the use of a 5 micrograms disc had little effect on these results, but a significant number of MRSA strains produced inhibition zones when disc diffusion tests were incubated at 35 degrees C, and many sensitive strains showed scanty growth on salt-containing agar at high methicillin concentrations in agar dilution tests . Iso-Sensitest agar did not appear to support the growth of the minority resistant populations of MRSA unless supplemented with menadione and thiamine, and even with supplemented Iso-Sensitest medium a few presumptively resistant strains appeared methicillin-sensitive in both disc diffusion and agar dilution tests.

J Antimicrob Chemother, 1987 Oct, 20(4), 595 - 7
Ciprofloxacin treatment of Staphylococcus aureus infections; Righter J; Ciprofloxacin appears to be safe and effective for a wide variety of clinical infections . In-vitro and animal studies point to high cure rates for both methicillin-sensitive and methicillin-resistant Staphylococcus aureus infections . Seventeen patients with staphylococcal infections severe enough to require hospital admission and initial parenteral therapy were treated with ciprofloxacin; the results were poor, with clinical failure in five and bacteriological failure in 12 . All pathogens isolated were susceptible to ciprofloxacin both before and after therapy and tolerance was not detected . Further study is required before ciprofloxacin can be recommended for life threatening staphylococcal infections.

Epidemiol Infect, 1987 Oct, 99(2), 349 - 53
Staphylococcal food poisoning on a cruise ship; Waterman SH et al.; Two waves of vomiting and/or diarrhoea affected approximately 215 of the 715 passengers on a Caribbean cruise ship . The outbreak was independently associated with eating cream-filled pastries at two separate meals . Staphylococcus aureus phage type 85/+ was isolated from cases and pastry cooks, but not from controls . This is the first well-documented outbreak of staphylococcal food poisoning on a cruise ship.

Am J Vet Res, 1987 Oct, 48(10), 1456 - 60
In vitro sensitization and stimulation of bovine lymphocytes with encapsulated or nonencapsulated Smith strain of Staphylococcus aureus; Ojo-Amaize EA et al.; An in vitro immunization procedure was developed, whereby bovine blood and mammary gland lymphocytes were isolated, and their blastogenic responses (measured by {3H}thymidine uptake) to sensitization and subsequent stimulation with selected antigens were determined . Capsular extracts of Staphylococcus aureus, encapsulated S aureus, and nonencapsulated S aureus were used as test antigens . Differences were observed between kinetics of the secondary response to encapsulated and nonencapsulated S aureus . The secondary response to nonencapsulated S aureus peaked in 48 hours, whereas the secondary response to the encapsulated S aureus peaked in 72 hours . The delayed peak response to encapsulated S aureus was observed only when encapsulated S aureus was used for sensitization of lymphocytes, regardless of whether the encapsulated or nonencapsulated strain was used for stimulation . Sensitization of lymphocytes with the nonencapsulated strain and stimulation with the encapsulated strain did not alter the kinetics of the secondary response of cells sensitized with the nonencapsulated strain . Seemingly, T and B cells were responsive to in vitro immunization with S aureus antigens.

Am J Med, 1987 Oct, 83(4), 801 - 3
Staphylococcus aureus septicemia mimicking fulminant Rocky Mountain spotted fever; Milunski MR et al.; Septicemia due to Staphylococcus aureus can be a difficult diagnosis to make early in its presentation . This report illustrates a case that mimicked fulminant Rocky Mountain spotted fever in a patient with no other medical problems that might predispose her to the development of staphylococcal sepsis . Epidemiology, cerebrospinal fluid characteristics, and early biopsy of skin lesions are emphasized as important factors leading to early diagnosis and definitive treatment.

J Clin Microbiol, 1987 Oct, 25(10), 1956 - 61
Isolation of Staphylococcus aureus L forms from experimentally induced bovine mastitis; Owens WE; Bacterial L forms were isolated from milk samples of dairy cattle infected experimentally with Staphylococcus aureus . Initially, bacterial L forms were induced in vitro from 12 of 44 S . aureus strains isolated from bovine mastitis . Cows were experimentally infected in two experiments with strains shown in vitro to be easily inducible to L form and with S . aureus Newbould 305 . Each quarter of the mammary gland was infected with either 10(3) or 10(6) CFU of the test strains . Treatment was initiated with 100,000 U of penicillin G per quarter at the first signs of clinical mastitis . Milk samples were collected daily and cultured on bovine blood agar and PPLO agar (Difco Laboratories, Detroit, Mich.) with 10% horse serum and 5% NaCl . Staphylococcal L forms were isolated from milk samples collected from infected glands in both experiments after antibiotic therapy . Glands with the highest concentrations of leukocytes and bacteria were most likely to yield L forms in milk samples after treatment was initiated . Cows harboring L forms typically yielded parental organisms after cessation of antibiotic therapy . No detectable changes occurred in antibiotic susceptibilities, coagulase production, or biochemical activities in strains induced to L form followed by reversion to the parental form . These results demonstrated that L forms can occur during treatment of bovine mastitis and that L forms may be one explanation for the poor response of staphylococcal bovine mastitis to antibiotic therapy.

J Clin Microbiol, 1987 Oct, 25(10), 1932 - 3
Predominance of capsular polysaccharide type 5 among oxacillin-resistant Staphylococcus aureus; Fournier JM et al.; The relationship between capsular polysaccharide types 5 and 8 and resistance of Staphylococcus aureus to oxacillin was studied with a collection of 406 clinical isolates from six French hospitals . Of 175 type 5 isolates, 84 (48%) were resistant to oxacillin . In contrast, only 8 of 160 type 8 isolates (5%) and 5 of 71 nontypeable isolates (7%) were resistant to oxacillin . Therefore, capsular typing of clinical isolates of S . aureus may facilitate the choice of first-line antibiotic therapy.

Am J Kidney Dis, 1987 Oct, 10(4), 281 - 6
A five-year study of the microbiologic results of exit site infections and peritonitis in continuous ambulatory peritoneal dialysis; Piraino B et al.; We studied the culture results from 321 continuous ambulatory peritoneal dialysis (CAPD) related infections (exit site, tunnel infections, and peritonitis) in 137 patients over a 5-year period to determine the contribution of exit site and tunnel infections to peritonitis and catheter loss . Seventeen percent of peritonitis episodes were associated temporally and by microbiologic results with exit site or tunnel infections . Twenty-one percent of exit site and tunnel infections and 20% of peritonitis episodes resulted in catheter loss . Peritonitis due to Staphylococcus aureus was more likely to be associated with an exit site or tunnel infection and was more likely to result in loss of the catheter than peritonitis due to Staphylococcus epidermidis . Peritonitis and exit site infections due to Pseudomonas sp also frequently resulted in catheter removal . We found that exit site infections cause significant morbidity in CAPD patients . Further studies in this area are needed.

Surg Gynecol Obstet, 1987 Oct, 165(4), 339 - 42
Comparison of nosocomial infections due to Staphylococcus aureus and enterococci in a general hospital; Leoung GS et al.; Hospital acquired infections caused by enterococci are increasing in incidence . This increase has been attributed, in part, to widespread use of cephalosporin antibiotics which lack activity against enterococci . To test this hypothesis, we compared patients having nosocomial enterococcal pulmonary and wound infections with those patients having nosocomial staphylococcal infections for antecedant cephalosporin use . In a six month period, we found 14 instances of nosocomial enterococcal wound and respiratory infections; 13 were superinfections (occurring during or up to one week after administration of antimicrobials) . Seven had coexisting pathogens noted . In the same six month period, 30 instances of Staphylococcus aureus nosocomial respiratory tract and wound infections were found and only 13 were superinfections (p less than 0.002 compared with enterococcal superinfections) . Nine had coexisting pathogens noted . The mean age, gender distribution and underlying illnesses in the Staphylococcus aureus and enterococcal superinfection groups were comparable . Of the 14 instances of enterococcal nosocomial infections, 11 were associated with administration of a cephalosporin compared with seven of 30 for staphylococcal infections (p less than 0.002) . Nosocomial enterococcal infections are commonly associated with antimicrobial therapy and the use of cephalosporins may selectively predispose patients to increased risk of enterococcal superinfections.

J Pediatr, 1987 Oct, 111(4), 513 - 8
Five years' experience with continuous ambulatory or continuous cycling peritoneal dialysis in children; von Lilien T et al.; In 93 children, end-stage renal disease was treated with the new dialytic methods of continuous ambulatory peritoneal dialysis (CAPD) or continuous cycling peritoneal dialysis (CCPD) over 5 years . Modality survival rates at 36 months with CAPD, CCPD, or both were 20%, 93%, and 87%, respectively . Use of CCPD as the primary dilaytic method increased during the study period . The peritonitis rate was one episode per 11.8 patient treatment months and was similar with both CAPD and CCPD . Gram-positive organisms were cultured in 34% of these episodes of peritonitis . Staphylococcus aureus peritonitis was associated with a recurrence rate of 40% and led to catheter replacement in 45% of the episodes . Peritoneal membrane failure necessitating switching to hemodialysis was related to peritonitis in three patients . Of the 74 peritoneal catheters that required replacement, 70% were infected . Serial serum levels of urea nitrogen, potassium, calcium, phosphorus, albumin, and alkaline phosphatase remained stable, whereas serum creatinine level rose slightly over time . Episodes of hyperkalemia, hypercalcemia, and hyperphosphatemia were observed at a frequency of one episode per 12.2, 4.6, and 2.5 treatment months, respectively . Blood transfusions were required in once per 1.5 and 3.3 treatment months in seven anephric patients and in 35 patients with their own kidneys, respectively (P = 0.05) . In prepubertal patients who received CAPD or CCPD for greater than 1 year, little or no improvement in growth occurred in relationship to either chronologic or bone age.

J Immunol, 1987 Oct 1, 139(7), 2237 - 41
Prominent IgM rheumatoid factor production by human cord blood lymphocytes stimulated in vitro with Staphylococcus aureus Cowan I; Levinson AI et al.; We previously reported that Staphylococcus aureus Cowan I (SAC) is a potent stimulant of IgM rheumatoid factor (RF) production by normal adult peripheral blood mononuclear cells . In the current study, we compared the capacity of normal adult peripheral blood mononuclear cells and cord blood mononuclear cells to produce IgM RF . Although both populations of cells consistently produced IgM RF in response to SAC, the quantity of RF produced by cord blood cells (128 +/- 18 ng/ml, mean +/- SEM) greatly exceeded that of adult cells (37 +/- 5 ng/ml, p less than 0.0005) even though both populations of cells demonstrated comparable total IgM responses . Remarkably, 16.7% of total IgM produced by cord blood cells in response to SAC showed RF activity compared to only 3.4% (p less than 0.0001) of the total IgM produced by SAC-stimulated adult cells . Thus, precursors of IgM RF-secreting cells are not only a consistent feature of the normal adult human B cell repertoire, but they are especially represented at the time of birth . These findings are consistent with the hypothesis that IgM RF originates from germ line genes and underscore the utility of SAC as a probe for analyzing the production of this autoantibody.

Infect Immun, 1987 Oct, 55(10), 2332 - 40
Immunosuppressive effects of Centipeda periodontii: selective cytotoxicity for lymphocytes and monocytes; Shenker BJ et al.; We have examined soluble sonic extracts prepared from several strains of Centipeda periodontii for their ability to alter human lymphocyte function . These organisms were isolated from subgingival plaque of patients with periodontal disease . We found that sonicates from several, but not all, strains of C . periodontii caused a dose-dependent inhibition of lymphocyte responsiveness to concanavalin A, phytohemagglutinin, pokeweed mitogen, and formalinized Staphylococcus aureus . Inhibition was associated with a concomitant decrease in cell viability assessed by trypan blue exclusion, 51Cr release, and electron microscopy . The maximal number of dead cells was observed 20 to 24 h after exposure to the sonic extract . Susceptible cells include human lymphocytes (both B and T), monocytes, and erythrocytes, whereas polymorphonuclear cells, murine L-929 fibroblasts, and sheep erythrocytes were not affected . Preliminary characterization of the cytotoxic activity indicates that it is heat labile and trypsin sensitive and has an Mr of 60,000 . It has been proposed that impaired host defense may play a pivotal role in the pathogenesis of periodontal diseases . The data presented in this paper suggest that immunosuppression (local or systemic or both) could be initiated by C . periodontii . This immunosuppression may alter the nature and consequences of host-parasite interactions, thereby enhancing the pathogenicity of C . periodontii itself or some other opportunistic organism.

J Infect Dis, 1987 Oct, 156(4), 597 - 606
Neutrophil uptake of vaccinia virus in vitro; West BC et al.; We studied human neutrophils for uptake of vaccinia virus . Uptake was determined radiometrically and by electron microscopy . Vaccinia virus was labeled with 14C or 3H, incubated with neutrophils, and quantified in neutrophil pellets in a new radiometric phagocytosis assay . Better results were obtained from assays of {3H}thymidine-labeled virus; uptake increased through 1 hr and then plateaued . Phagocytosis of 3H-labeled Staphylococcus aureus was normal . Uptake of virus was serum dependent . Hexose monophosphate shunt activity was measured by two methods . No 14CO2 from {14C}1-glucose accompanied uptake of vaccinia virus, in contrast to the respiratory burst accompanying bacterial phagocytosis . Electron microscopy showed intact to slightly digested intraphagolysosomal vaccinia virus . Pock reduction assay showed a decrease in viral content due to neutrophils until 6 hr of incubation, when a modest but significant increase was observed . Thus, neutrophil uptake of vaccinia virus is distinguished from bacterial phagocytosis.

J Rheumatol, 1987 Oct, 14(5), 873 - 9
Synovial fluids from patients with rheumatoid arthritis possessed B cell differentiation activity contributing to synthesis of rheumatoid factor in vitro; Kitani A et al.; B cell differentiation activity assayed on B cells preactivated by Staphylococcus aureus Cowan I was found in synovial fluids (SF) of patients with rheumatoid arthritis (RA) (n = 14), while little, if any, activity was found in SF of patients without RA (n = 13) including patients with osteoarthritis and traumatic arthritis . There was a positive correlation between the RA hemagglutination assay titer and B cell differentiation activity (r = 0.7438) . This molecule with molecular weight ranges 15-20 kDa as well as supernatant from phytohemaglutinin stimulated normal T cells could promote not only polyvalent immunoglobulin synthesis but also rheumatoid factor synthesis by lymphocytes from patients with RA in vitro, and was considered to be distinct from interleukin-1, interleukin-2, interferon r, or proteolytic lysosomal enzymes which have been shown to modulate B cell activation.

Biol Chem Hoppe Seyler, 1987 Oct, 368(10), 1327 - 31
Structural studies on the murine granulocyte colony-stimulating factor; Simpson RJ et al.; Granulocyte-colony stimulating factor (G-CSF) is a glycoprotein hemopoietic growth factor which regulates the production of granulocytes and macrophages . Reversed-phase microbore high-performance liquid chromatography was employed to purify a number of tryptic and Staphylococcus aureus V8 proteinase peptides generated from approximately 400 pmol G-CSF purified from medium conditioned by lungs from mice previously injected with endotoxin . N-Terminal amino-acid sequence analyses were performed on the parent polypeptide and on four tryptic peptides and one Staphylococcus aureus V8 protease peptide, yielding 68 unique amino-acid assignments; this corresponds to approximately 38% of the molecule.

Epidemiol Infect, 1987 Oct, 99(2), 343 - 7
Pattern of antibiotic and heavy-metal ion resistance in recent hospital isolates of Staphylococcus aureus; Millar MR et al.; Two hundred and one strains of Staphylococcus aureus isolated from in-patients and out-patients were examined for sensitivity to antibiotics, heavy-metal ions and ethidium bromide and for phage-typing pattern . Heavy-metal ion resistance was less frequent than reported in previous studies and was as frequent in penicillinase non-producing as producing strains . 'Methicillin-resistant' strains were resistance to ethidium bromide and mercury . Resistance to heavy-metal ions, including cadmium, may be becoming less common amongst clinical isolates of S . aureus.

Am J Clin Nutr, 1987 Oct, 46(4), 659 - 64
Pyridoxine supplementation: effect on lymphocyte responses in elderly persons; Talbott MC et al.; The effect of pyridoxine supplementation on lymphocyte responsiveness was investigated in 15 persons aged 65-81 y . Eleven subjects received 50 mg/d pyridoxine HCl (PN) . Four subjects received a placebo . Lymphocyte proliferation to T and B cell mitogens, lymphocyte subpopulations with monoclonal antibodies, and plasma pyridoxal 5'-phosphate (PLP) were measured before and after 1 and 2 mo of supplementation . After 1 and 2 mo plasma PLP levels increased by 195 +/- 88 nM and 201 +/- 84 nM, respectively, in subjects receiving PN . With PN supplementation, lymphocyte proliferation increased significantly in response to phytohemagglutinin (p less than 0.01), pokeweed mitogen (p less than 0.01), and Staphylococcus aureus (Cowain I) (p less than 0.05) . For PN-treated subjects with low presupplement plasma PLP levels, lymphocyte blastogenesis also increased significantly (p less than 0.01) in response to concanavalin A . Percentages of T3+ and T4+ but not T8+ cells increased significantly (p less than 0.05) in PN-treated subjects . These results suggest that improving vitamin B-6 status is important in stimulating immunocompetence in the elderly.

Mol Cell Biochem, 1987 Oct, 77(2), 179 - 85
Iodination of the progesterone receptor from hen oviduct spares the DNA-binding domain; Fahnestock M; The progesterone receptor from hen oviduct is isolated as a complex of two subunits, A and B . The A protein binds one molecule of progesterone and also binds to DNA with high affinity . The native A protein can be labeled with iodine with no loss of DNA-binding activity . Limited Staphylococcus aureus V8 protease digestion of the labeled preparation results in a number of DNA-binding and non-DNA-binding fragments of the receptor . The progesterone-binding domain contains iodine label . However, two low-molecular-weight DNA-binding fragments do not contain iodine label, indicating a lack of susceptible tyrosine residues near the DNA-binding site of the native receptor . The labeled receptor and its fragments will facilitate studies of the isolated DNA-binding and progesterone-binding domains of the hen A protein as well as of the activity of the native receptor in the presence and absence of hormone.

J Rheumatol, 1987 Oct, 14(5), 991 - 4
Primary septic arthritis in heroin users: early diagnosis by radioisotopic imaging and geographic variations in the causative agents; Lopez-Longo FJ et al.; We reviewed 37 cases of septic arthritis in heroin users . Our data confirm the predominance of the fibrocartilaginous joint infections in this group (sacroiliac joint 39%, chondrosternocostal unions 37%) . In Spain, Staphylococcus aureus is the most commonly isolated organism (73%) . This emphasizes the geographic variations in the causative germs since, in contrast to other reports, we have not identified any gram negative bacillary arthritis in our population of heroin users . Our data show that the 67gallium citrate scintigraphy is positive earlier than the 99mTc-MDP bone scan in the poorly vascularized joints (p less than 0.0005) . The early localization of the infectious focus by 67gallium citrate scintigraphy followed by a prompt bacteriologic diagnosis (blood, synovial fluid or tissue cultures) allowed good therapeutic results.

Proc Natl Acad Sci U S A, 1987 Oct, 84(20), 7014 - 8
Conformational studies of alpha-globin in 1-propanol: propensity of the alcohol to limit the sites of proteolytic cleavage; Iyer KS et al.; Selective condensation of the unprotected fragments of alpha-globin--namely, alpha 1-30 and alpha 31-141--is catalyzed by Staphylococcus aureus V8 protease in the presence of 25% 1-propanol . The propensity of 1-propanol to induce the alpha-helical conformation and to generate a "native-like" topology for the polypeptide chain has been now investigated in an attempt to understand the molecular basis of this enzyme-catalyzed stereospecific condensation . Removal of heme from the alpha-chain decreases the overall alpha-helical conformation of the protein considerably . A significant amount of the alpha-helical conformation is restored in the presence of 25% 1-propanol and the digestion of alpha-globin by V8 protease becomes more selective concomitant with the increase in helicity . V8 protease digestion of alpha-globin at pH 6.0 and 4 degrees C occurs at Glu-30, Asp-47, Glu-27, and Glu-23 in the absence of 1-propanol . In the presence of 25% 1-propanol, the digestion is selective to the peptide bond of Glu-30 . This selectivity appears to be a characteristic feature of the native conformation of alpha-chain (polypeptide chain with bound heme) . 1-Propanol induces the alpha-helical conformation into RNase S peptide also . However, this increased helical conformation did not protect the RNase S peptide from V8 protease digestion at the Glu-9-Arg-10 peptide bond . RNase S peptide is an alpha-helical conformation in RNase S, an interacting fragment-complementing system of S protein and S peptide . S peptide is resistant to V8 protease hydrolysis in this conformation . Thus, the resistance of a peptide bond in a segment of a protein to protease digestion appears to be a consequence of the secondary structure as well as the tertiary interactions of this segment with the rest of the molecule . The results suggest that the 1-propanol induces alpha-helical conformation into segments of alpha-globin as well as packing of these helices in a native-like topology.

Int J Pediatr Nephrol, 1987 Oct-Dec, 8(4), 231 - 4
Shunt nephritis with crescent formation; Toth T et al.; The case of a 17-year-old girl is reported, who presented shunt nephritis, 8 years after the ventriculo-atrial shunt insertion was made . Renal biopsy showed membranoproliferative glomerulonephritis . The symptoms of uremia mingled with those of chronic septic state . Persistent hypocomplementemia and cryoglobulinemia dominated the picture of the disease . At the beginning of the disease Staphylococcus aureus abunded in the blood culture . In spite of the therapy the general status of the patient declined steadily.

Br J Dis Chest, 1987 Oct, 81(4), 404 - 6
An unusual manifestation of staphylococcal lung infection; Scammell AM et al.; An infant developed a rapidly progressive respiratory illness with cystic lesions on chest radiography . At thoracotomy there were multiple bullae on the surface of the lung . Staphylococcus aureus, of phage type 71, was isolated from the bullous fluid . We believe the lung disease was caused by an epidermolytic toxin.

Cornell Vet, 1987 Oct, 77(4), 293 - 302
Evaluation by enzyme-linked immunosorbent assay of local and systemic production of milk immunoglobulins to surface exopolysaccharide antigen in cows with staphylococcal mastitis; Loeffler DA et al.; An enzyme-linked immunosorbent assay (ELISA) was developed to evaluate milk immunoglobulin levels to surface exopolysaccharide antigen of Staphylococcus aureus in cows with staphylococcal mastitis . Quarter milk samples were obtained from 24 lactating dairy cows on two occasions, one month apart . Cows were classified as S . aureus-positive (S . aureus cultured from at least one quarter on both sample dates) or S . aureus-negative . Individual quarter samples were tested for IgA (representing local synthesis) and IgG1 (primarily of systemic origin) specific for staphylococcal surface exopolysaccharide antigen . No significant differences were found for specific IgA or IgG1 between S . aureus-positive and S . aureus-negative cows, nor between infected and non-infected quarters of S . aureus-positive cows . The data indicate that, in cows with staphylococcal mastitis, milk immunoglobulins specific for exopolysaccharide antigen are not significantly increased by either the systemic or the local immune response.

Eur J Clin Microbiol, 1987 Oct, 6(5), 589 - 91
Comparison of seven coagulase tests for identification of Staphylococcus aureus; Weers-Pothoff G et al.; Four coagglutination tests for the identification of Staphylococcus aureus were compared with the ordinary slide and tube coagulation tests using two groups of staphylococcal strains (one isolated in the authors' laboratory and the other identified by a reference laboratory) . The correct identification rate in the two groups was respectively for slide test 96.9% and 92.7%, tube coagulation (citrate plasma) 99.0% and 94.8%, tube coagulation (EDTA plasma) 99.0% and 91.7%, API Staphase III 100% and 91.7%, Staph Rapid 97.9% and 91.3%, Staphyslide 99.0% and 94.6%, Staphaurex 96.9% and 92.7% . The sensitivity of Staph Rapid, Staphyslide and Staphaurex was slightly higher than that of the other tests . These three tests and the slide test were considerably more rapid as regards identification than the other tests.

Acta Orthop Scand, 1987 Oct, 58(5), 529 - 34
Hematogenous infection after knee arthroplasty; Bengtson S et al.; Twenty-five hematogenously infected knee arthroplasties in 20 patients (17 with rheumatoid arthritis and 3 with arthrosis) were followed for 3 years . Staphylococcus aureus was the major infecting organism . Three patients with four arthroplasties died of sepsis . Two patients had removal of the arthroplasty, one of which resulted in an above-the-knee amputation . Four out of five arthrodeses fused . Two knees healed after early debridement and two healed without surgery . Ten knees had successful revision arthroplasty . Rheumatoid arthritis and constrained prostheses increase the risk of hematogenous infection . Any infection and especially cutaneous lesions in a patient with a knee arthroplasty should be treated vigorously.

Arch Pathol Lab Med, 1987 Oct, 111(10), 983 - 7
Causes of death in renal transplant recipients . A review of autopsy findings from 1966 through 1985; Scroggs MW et al.; From 1966 through 1985, a total of 640 patients received 739 renal transplants at a single center transplantation program . Of 245 total deaths, a slide and chart review of all 116 autopsied cases (47%) identified the major causes of death as pneumonia (n = 43), sepsis (n = 32), hemorrhage (n = 15), peritonitis (n = 11), meningitis (n = 7), and pulmonary embolism (n = 5) . Eighty-five (73.3%) of these patients died of complications directly associated with immunosuppression, almost all (n = 82) as a result of infection . Organisms most frequently identified at death were gram-negative bacilli (n = 72), Candida species (n = 23), cytomegalovirus (n = 17), enterococcus (n = 14), Staphylococcus aureus (n = 11), Aspergillus species (n = 10), Pneumocystis carinii (n = 5), and mycobacteria (n = 5) . Significant associations were found between bolus steroid antirejection therapy and infection with Aspergillus cytomegalovirus . Diabetics had a higher incidence of fungal infections and bowel perforation than nondiabetics . During this 20-year period, overall one-year actual patient survival rates for the four respective five-year intervals increased dramatically (69.9%, 68.2%, 83.3%, and 91.8%), but the normalized death rate showed a smaller decrease for infectious vs noninfectious causes.

J Antimicrob Chemother, 1987 Oct, 20(4), 505 - 11
In-vitro activity of LY146032 against Staphylococcus aureus and S . epidermidis; Coudron PE et al.; The antibacterial activity of LY146032, a new cyclic polypeptide, was compared with that of vancomycin against 163 isolates of methicillin-susceptible and methicillin-resistant Staphylococcus aureus and S . epidermidis . The MICs of LY146032 and vancomycin for 90% of the isolates were 1.0 mg/l and 2.0-4.0 mg/l, respectively . Interpretative guidelines for 15 micrograms and 30 micrograms LY146032 discs could not be established because all isolates tested were susceptible to LY146032, on the basis of achievable serum levels . With one exception, MBC to MIC ratios of LY146032 and vancomycin were less than or equal to 2 . Inoculum size had minimal effect on ratios . In time-kill studies, both drugs were 99.9% bactericidal at 6 h for four isolates at concentrations four times the MBC or less . No change in LY146032 activity was observed between isogenic pairs that were resistant to other antibiotics . Thus, LY146032 was very active in vitro against all staphylococcal isolates.

Eur J Biochem, 1987 Oct 1, 168(1), 141 - 51
Rat glycine methyltransferase . Complete amino acid sequence deduced from a cDNA clone and characterization of the genomic DNA; Ogawa H et al.; The amino terminus of glycine methyltransferase from rat liver is blocked . A hexapeptide containing the blocked amino-terminal residue was obtained from a tryptic digest of the purified enzyme and its amino acid sequence was determined to be Ac-Val-Asp-Ser-Val-Tyr-Arg by Edman degradation and fast-atom-bombardment mass spectrometry after fragmentation with Staphylococcus aureus protease V8 . A full-length cDNA clone for the enzyme was isolated from a lambda gt11 rat liver cDNA library using the previously obtained pGMT A56 cDNA {Ogawa, H., Gomi, T., Horii, T., Ogawa, H . & Fujioka, M . (1984) Biochem . Biophys . Res . Commun . 124, 44-50} as a probe . The amino acid sequence deduced from the nucleotide sequence contained both amino- and carboxyl-terminal sequences . The predicted amino acid composition and molecular mass were also in agreement with the published data obtained with the purified protein . Five clones for the glycine methyltransferase gene were isolated from a Charon 4A library containing EcoRI digest of rat liver DNA by in situ plaque hybridization . All clones had inserts of 6500 base pairs, consistent with the size of EcoRI genomic DNA fragment determined by Southern blot hybridization . Sequence analysis of a 5400-bp fragment of the insert DNA lacking a 1100-bp 5' region and comparison of the sequence with that of the cDNA showed that the insert DNA entirely encoded glycine methyltransferase and the gene consisted of six exons and five introns . S1 nuclease protection mapping and primer extension analysis allowed us to propose that the A residue located 19 bp upstream from the translation initiation codon is the site of transcription initiation . TATA, CAAT and GC sequences, and the complementary sequence to the enhancer core element, were located upstream of the transcription initiation site.

Infect Immun, 1987 Oct, 55(10), 2398 - 403
Oxidative response of human neutrophils, monocytes, and alveolar macrophages induced by unopsonized surface-adherent Staphylococcus aureus; Devalon ML et al.; In contrast to results with bacterial suspensions, phagocytosis of unopsonized bacteria readily occurs when bacteria are adhered to glass or plastic surfaces . However, in contrast to neutrophils, alveolar macrophages produced much less DNA denaturation as measured by acridine orange metachromasia of phagocytized Staphylococcus aureus . We have studied the phagocytosis of unopsonized surface-adherent S . aureus and the subsequent production of reactive oxygen species by peripheral blood neutrophils, monocytes, and alveolar macrophages . Phagocyte-free systems were then used to show the relationship of the reactive oxygen species produced by neutrophils and alveolar macrophages and the denaturation of unopsonized S . aureus DNA with acridine orange . Peripheral blood neutrophils, monocytes, and alveolar macrophages from normal human volunteers were added to vials with adherent S . aureus without opsonin . Bacterial uptake and luminol- and lucigenin-dependent chemiluminescence were measured . Neutrophils developed much greater luminol-dependent chemiluminescence than monocytes or alveolar macrophages . Compared with neutrophils and monocytes, alveolar macrophages developed significantly greater concentrations of superoxide, as measured by lucigenin-dependent chemiluminescence and ferricytochrome c reduction . These findings suggested that products of the myeloperoxidase-hydrogen peroxide-halide pathway were generated when peripheral blood neutrophils were stimulated and that alveolar macrophages primarily produced superoxide . When these reactive oxygen species were generated in phagocyte-free systems containing S . aureus, products of the myeloperoxidase-hydrogen peroxide-halide pathway produced denaturation of S . aureus DNA, whereas superoxide did not . Thus, differences in reactive oxygen species produced during phagocytosis may be related to the different capacities of neutrophils and alveolar macrophages to denature unopsonized adherent S . aureus DNA.

Microb Pathog, 1987 Oct, 3(4), 299 - 307
Identification of conserved regions for species and subspecies specific epitopes on the major outer membrane protein of Chlamydia trachomatis; Ma JJ et al.; Proteolytic peptides containing the serological epitopes present on the major outer membrane protein of Chlamydia trachomatis were studied by immunoblots . Monoclonal antibodies which have been defined by micro-immunofluorescence typing as serovar-, subspecies and species-specific were utilized . The reactivity of either serovar-specific or species-specific monoclonal antibodies in the immunoblots was similar to that in the micro-immunofluorescence test . However, monoclonal antibodies which demonstrated subspecies-specific reactivity by the micro-immunofluorescence test expressed species-specific reactivity by the immunoblot method . The serovar-specific epitope which was identified on the major outer membrane protein was lost after proteolytic digestion with Staphylococcus aureus V8 protease . Immunoblots of the proteolytic fragments identified the species-specific epitope on several peptides and the subspecies-specific epitope on fewer peptides than those in which the species-specific epitope was identified . In addition, both the species and subspecies epitopes were present on the same fragments of different serovars . The data indicate that the species epitope is on conserved regions of the major outer membrane protein and is structurally clustered with a subspecies-specific epitope.

Immunol Cell Biol, 1987 Oct, 65 Pt 5, 411 - 7
Effect of dextran sulphate on IgG subclass of antibody in efferent popliteal lymph of sheep; Kerlin RL et al.; IgG1 and IgG2 participation in anti-hapten, anti-carrier and immunoglobulin-containing cell (Ig-cc) responses was studied in sheep immunised with killed Staphylococcus aureus-Dinitrophenyl (DNP) conjugates . The antigen was given with or without the polyanionic adjuvant dextran sulphate (DXS) . Animals were immunised intracutaneously on the lateral hock of one hind leg with 1 X 10(9) S . aureus-DNP with or without 20 mg DXS . Six weeks later, primed sheep underwent surgery to cannulate an efferent popliteal lymphatic vessel in the immunised leg . Forty-eight hours after surgery, each sheep was given a further intracutaneous injection of 1 X 10(9) S . aureus-DNP with or without 20 mg DXS adjacent to the primary injection site . IgG1-cc and IgG2-cc in lymph were counted using indirect immunofluorescence . Total IgG1 and IgG2 anti-staphylococcal and anti-DNP antibody output in lymph was determined using enzyme-linked immunosorbent assays (ELISA's) . DXS markedly increased antibody responses to S . aureus-DNP in the popliteal lymph node of sheep and was shown to strongly enhance immunological memory . The ratios of IgG2-cc:IgG1-cc and IgG2:IgG1 anti-staphylococcal antibody in lymph-draining popliteal lymph nodes of sheep stimulated with DXS and S . aureus-DNP was significantly greater than the same ratios in animals given antigen alone . The IgG subclass-specific immunomodulatory effects of DXS were exerted whether the adjuvant was given with primary and/or secondary inoculations . There was no difference in the ratio of IgG2:IgG1 anti-DNP antibody in lymph from animals given S . aureus-DNP with and without DXS.

Biochim Biophys Acta, 1987 Sep 25, 921(2), 370 - 7
Positional specificity and substrate preference of purified Staphylococcus aureus lipase; Rollof J et al.; We have studied the substrate preference and specificity, including positional specificity, of a lipase purified from Staphylococcus aureus (strain FN 37) . This extracellular bacterial enzyme is relatively insensitive to product inhibition, and hydrolyzes tri-, di- and monooleoylglycerol in emulsified and micellar form at similar rates and without marked substrate preference . The lipase lacks positional specificity, and the hydrolysis of triacylglycerol proceeds rapidly to free fatty acid and glycerol without accumulation of intermediary products.

Biochim Biophys Acta, 1987 Sep 25, 921(2), 364 - 9
Purification and characterization of a lipase from Staphylococcus aureus; Rollof J et al.; An extracellular lipase from Staphylococcus aureus (strain FN 37) was purified to homogeneity . A cell-free culture broth was subjected to ammonium sulphate precipitation, and the lipase was isolated from the resuspended pellet by adsorption chromatography on octyl-Sepharose . The purification was 957-fold, and the recovery of the octyl-Sepharose chromatography was about 100% . The specific activity of the purified lipase was 546 mU of lipase activity per micrograms protein . The purity of the final product was documented by SDS-polyacrylamide gel electrophoresis in which a homogeneous protein band of 43 kDa was found . In gel chromatography on Sephadex G-200 the lipase eluted as a homogeneous peak with an apparent molecular mass of 110 kDa, suggesting that the lipase may exist as an oligomer in physiological media . Analysis of the amino-acid composition revealed a predominance of polar, non-charged amino acids, with serine accounting for 24 mol% of the amino-acid residues.

J Am Vet Med Assoc, 1987 Sep 15, 191(6), 681 - 4
Isolation of L-form variants after antibiotic treatment in Staphylococcus aureus bovine mastitis; Sears PM et al.; An unstable L-form of Staphylococcus aureus was identified in milk samples from 3 quarters of 2 cows after treatment with cloxacillin . Milk samples incubated on standard 5% blood agar plates were culture-negative for 7 to 30 days after treatment, but S aureus was reisolated in 80% of 66 samples by additional culturing on enriched L-form media when incubated in 10% CO2 at 37 C . The organism was identified at various phases of reversion of L-form agar and was confirmed on transfer to blood agar plates.

J Biol Chem, 1987 Sep 15, 262(26), 12768 - 79
Identification of two testosterone-responsive testicular proteins in Sertoli cell-enriched culture medium whose secretion is suppressed by cells of the intact seminiferous tubule; Cheng CY et al.; Of 30 proteins identified in medium from primary Sertoli cell-enriched cultures, five of these proteins appear to increase primarily in response to testosterone . Using high performance liquid chromatography, two of these proteins, designated CMB-22 and CMB-23, were purified to apparent homogeneity from medium . Both are monomeric proteins with apparent molecular weights of Mr 37,000 and Mr 40,000, respectively . Both interact with concanavalin A and wheat germ agglutinin on lectin blots . CMB-22 has a pI of 5.8; CMB-23 has two distinctive isoelectric variants with pIs of 5.4 and 5.2, the latter variant was designated CMB-23 Isoform . Polyvalent antisera raised against purified CMB-22 and CMB-23 in rabbits cross-reacted with one another . Removal of carbohydrate from these proteins by either enzymatic or chemical treatments reduced their apparent molecular weights but did not abolish their size differences on sodium dodecyl sulfate-polyacrylamide gels . Peptide maps generated by Staphylococcus aureus protease V8 and visualized by silver staining and immunoblots suggest that CMB-22 and CMB-23 are similar but distinctive proteins that share almost identical epitopes . A radioimmunoassay was developed and used to measure total immunoreactive CMB-22-like material (CMB-22 plus CMB-23 immunoreactivity) in culture media, biological fluids, and tissue extracts . The results of these studies showed that the secretion of CMB-22-like material is unique with regard to other proteins that have been identified in Sertoli cell-enriched cultures . That is, CMB-22-like material is secreted by Sertoli cell-enriched cultures, but cannot be detected in media from cultures of intact tubular segments in vitro . In addition, immunoreactive material is also not detected in the testicular fluids from interstitium, tubule, or rete testis . This is in striking contrast to other Sertoli cell proteins which are present in substantial concentrations in these fluids . These observations suggest that the secretion and possibly the hormone responsiveness of CMB-22 and CMB-23 are normally suppressed in the intact tubule both in vivo and in vitro . We propose that studies of CMB-22 and CMB-23 will provide important insights into cell-cell interactions in the seminiferous epithelium.

Biochem J, 1987 Sep 15, 246(3), 651 - 8
Nucleophilic re-activation of the PC1 beta-lactamase of Staphylococcus aureus and of the DD-peptidase of Streptomyces R61 after their inactivation by cephalosporins and cephamycins; Faraci WS et al.; It has been shown previously {Faraci & Pratt (1985) Biochemistry 24, 903-910; (1986) Biochemistry 25, 2934-2941; (1986) Biochem . J . 238, 309-312} that certain beta-lactam-processing enzymes form inert acyl-enzymes with cephems that possess good leaving groups at the C-3' position . These inert species arise by elimination of the leaving group from the initially formed and more rapidly hydrolysing acyl-enzyme, which has the 'normal' cephalosporoate structure . The present paper shows that a strong nucleophile, thiophenoxide, can catalyse the re-activation of three examples of these inert acyl-enzymes, generated on reaction of cephalothin and cefoxitin with the PC1 beta-lactamase of Staphylococcus aureus and of cephalothin with D-alanyl-D-alanine transpeptidase/carboxypeptidase of Streptomyces R61 . In view of the reversibility of the elimination reaction, demonstrated in model systems {Pratt & Faraci (1986) J . Am . Chem . Soc . 108, 5328-5333}, this catalysis is proposed to arise through nucleophilic addition to the exo-methylene carbon atom of the inert acyl-enzyme to regenerate a more rapidly hydrolysing normal cephalosporoate . Strong support for this scenario was obtained through comparison of the kinetics of the catalysed re-activation reaction with those of turnover of the relevant 3'-thiophenoxycephems, thiophenoxycephalothin and thiophenoxycefoxitin . The enzymes appear to stabilize the products of the elimination reaction with respect to the normal cephalosporoate, but more strongly to destabilize the transition states . The effects of other nucleophiles, including cysteine, glycine amide and imidazole, on the above enzymes and on other beta-lactamases can be understood in terms of the model reaction kinetics and thermodynamics.

J Immunol, 1987 Sep 15, 139(6), 1792 - 6
Anti-HLA class I antibodies inhibit the T cell-independent proliferation of human B lymphocytes; Taylor DS et al.; The role of major histocompatibility complex-encoded class I molecules in the proliferation of human B lymphocytes is presently unclear . This question was addressed by investigating the effect of three individually derived anti-HLA class I monoclonal antibodies (mAb) on purified human B cells (less than 1.5% T cells) stimulated by either the T-independent mitogen Staphylococcus aureus or the phorbol ester, phorbol-12-myristate-13-acetate . The three anti-HLA class I antibodies, whether specific for gene products of the HLA-A locus (mAb 131), HLA-B locus (mAb 4E), or HLA-A, -B, and -C locus (mAb W6/32), inhibited S . aureus-induced proliferation by 70 to 90% . This inhibition was significant over a 5-day culture period, was not altered by the addition of exogenous interleukin 2 or B cell growth factor, and was not due to nonspecific cytotoxicity . In addition, the inhibition of proliferation was unchanged when the mAb were added 12 hr after the initiation of culture . The proliferative response was not affected by either of the control antibodies OKB7 and R3-367 . In contrast with S . aureus-stimulated B cells, phorbol-12-myristate-13-acetate-induced proliferation was resistant to the inhibitory activity of HLA class I-specific antibodies . These results suggest that HLA class I molecules are involved in human B lymphocyte proliferation and may regulate a critical event preceding the upregulation of protein kinase C activity.

Biochem Biophys Res Commun, 1987 Sep 15, 147(2), 787 - 93
Evidence that bacteria induce translocation of protein kinase C activity in human polymorphonuclear leukocytes; Christiansen NO et al.; Particulate fraction associated protein kinase activity was studied in human polymorphonuclear leukocytes stimulated by bacteria . Staphylococcus aureus was found to increase particulate fraction associated protein kinase C activity in a time and concentration dependent manner . The increase comprised both the phospholipid dependent and independent kinase activity and was augmented by addition of serum . Similar observations were done using Staphylococcus epidermidis and Klebsiellae pneumoniae . However, Escherichia coli only increased the phospholipid independent kinase activity in the particulate fraction, which suggests the presence of protease activity.

Eur J Biochem, 1987 Sep 15, 167(3), 525 - 32
Characterization of epitopes on a variant surface glycoprotein from Trypanosoma congolense by six monoclonal antibodies; Reinwald E et al.; Monoclonal antibodies were isolated from mice immunized with variant surface glycoprotein of Trypanosoma congolense . Five out of the six monoclonals were able to detect epitopes at the cell surface in an indirect immunofluorescence analysis . One antibody did not react . Using protein-A-containing bacterial adsorbent all monoclonal antibodies precipitate glycosylated as well as non-glycosylated variant surface glycoprotein . Carbohydrate chains therefore do not appear to be part of the immunodeterminant structure recognized by the various monoclonal antibodies . Interaction of the monoclonal antibodies with protein fragments obtained by partial proteolysis with V8 protease from Staphylococcus aureus or papain allows the classification of the antibodies into three groups with different epitope specificity.

Biochim Biophys Acta, 1987 Sep 14, 930(2), 220 - 9
Identification of a phosphorylated form of phosphoenolpyruvate carboxykinase from the yeast Saccharomyces cerevisiae; Burlini N et al.; A phosphoprotein of 65 kDa, as determined by SDS-gel electrophoresis, has been isolated from yeast crude extracts . This phospho form copurifies with phosphoenolpyruvate carboxykinase in the enzyme purification procedure worked out in our laboratory (Tortora, P., Hanozet, G.M . and Guerritore, A . (1985) Anal . Biochem . 144, 179-185) . Moreover, both proteins bind strongly to 5'AMP-Sepharose 4B in the presence of Mn2+, whereas a substantially lower binding occurs if Mn2+ is replaced by Mg2+ . This binding pattern is consistent with the well-known Mn2+-dependence of yeast phosphoenolpyruvate carboxykinase . These data suggest that the 65-kDa protein might be a phosphorylation product of the native enzyme . Furthermore, although the phospho form is not immunoprecipitated by anti-phosphoenolpyruvate carboxykinase antibodies, addition of Protein A-Sepharose CL-4B to crude extracts preincubated with the antibodies results in the binding to the resin of the phospho form, thus providing immunological evidence for its identification as a modified form of native enzyme . The same 65-kDa phosphoprotein is detectable in extracts from cells grown in the presence of {32P}Pi, as well as in cell extracts incubated with {gamma-32P}ATP . Moreover, digestion of the phosphoprotein with BrCN or with Staphylococcus aureus V8 proteinase, yields two and three fragments, respectively, which appear parallel to digestion products of phosphoenolpyruvate carboxykinase, again supporting the proposed identification . Finally, analysis of the phosphorylated amino acids in the 65-kDa protein shows that phosphoserine is the only labelled phosphoamino acid.

J Biol Chem, 1987 Sep 5, 262(25), 11964 - 72
A heparin binding site in antithrombin III . Identification, purification, and amino acid sequence; Smith JW et al.; A heparin-binding peptide within antithrombin III (ATIII) was identified by digestion of ATIII with Staphylococcus aureus V8 protease followed by purification on reverse-phase high pressure liquid chromatography using a C-4 column matrix . The column fractions were assayed for their ability to bind heparin by ligand blotting with 125I-fluoresceinamine-heparin as previously described (Smith, J . W., and Knauer, D . J . (1987) Anal . Biochem . 160, 105-114) . This analysis identified at least three fractions with heparin binding ability of which the peptide eluting at 25.4 min gave the strongest signal . Amino acid sequence analysis of this peptide gave a partially split sequence which was consistent with regions encompassing amino acids 89-96 and 114-156 . These amino acids are present in a 1:1 molar ratio which is consistent with a disulfide linkage between Cys-95 and Cys-128 . High affinity heparin competed more effectively for the binding of 125I-fluoresceinamine-heparin to this peptide than low affinity heparin . Chondroitin sulfate did not block the binding of 125I-fluoresceinamine-heparin to the peptide . These data strongly suggest that the isolated peptide represents a native heparin-binding region within intact ATIII . Computer generation of a plot of running charge density of ATIII confirms that the region encompassing amino acid residues 123-141 has the highest positive charge density within the molecule . A hydropathy plot of ATIII was generated using a method similar to that of Kyte and Doolittle (Kyte, J., and Doolittle, R . F . (1982) J . Mol . Biol . 157, 105-132) . This plot indicates that amino acid residues 126-140 are exposed to the exterior surface of the molecule . Based on these data, we suggest that the region corresponding to amino acid residues 114-156 is a likely site for the physiological heparin-binding domain of ATIII . We also conclude that the proposed disulfide bridges within the protein are suspect and should be re-examined (Petersen, T . E., Dudek-Wojiechowska, G., Sottrup-Jensen, L., and Magnussun, S . (1979) in The Physiological Inhibitors of Coagulation and Fibrinolysis (Collen, D., Wiman, B., and Verstaeta, M., eds) pp . 43-54, Elsevier Scientific Publishing Co., Amsterdam).

Ophthalmology, 1987 Sep, 94(9), 1148 - 54
Bacterial scleral abscesses after retinal buckling operations . Pathogenesis, management, and laboratory investigations; Folk JC et al.; Between 1971 and 1985, 28 patients were treated for a scleral abscess after a scleral buckle procedure . Twenty-six of the patients had the original surgery performed between those years during which a total of 4480 buckling procedures were performed . Therefore, the incidence of scleral abscess after buckling procedures was 0.58% (26/4480) . All 28 patients had excessive pain, conjunctival chemosis and proptosis, a creamy white area of retina on the buckle, and a variable amount of vitreous inflammation . Seventeen of these patients had an especially severe scleral abscess with inflammatory vitreous opacification, which precluded visualization of the retinal vessels or allowed at most a hazy view of the optic nervehead or the scleral buckle . A review of these 17 severely involved patients showed that, in most cases (13 of 17), the vitreous inflammation was sterile . Therefore, most patients can be managed simply with immediate removal of the buckle along with topical and systemic antibiotics . A laboratory investigation showed that a cell-free millipore filtrate of Staphylococcus aureus culture in trypticase soy broth caused prominent but self-limited inflammation when injected intravitreally . Therefore, bacterial toxins or secretions could be the cause of the prominent vitreous inflammation in most patients with scleral abscesses . In another experiment, sponge exoplants, which had been soaked in a highly concentrated culture of S . aureus, were sutured episclerally in rabbits . A scleral abscess developed in 19 of 23 rabbit eyes with these infective sponges . Progressive scleral abscesses, which closely resembled those seen in humans, developed in 14 of the 23 eyes.(ABSTRACT TRUNCATED AT 250 WORDS)

J Antimicrob Chemother, 1987 Sep, 20(3), 399 - 404
Influence of various antibiotics on phagocytosis of Staphylococcus aureus by human polymorphonuclear leucocytes; Van der Auwera P et al.; The influence of several new antibiotics on phagocytosis of two strains of Staphylococcus aureus by human polymorphonuclear leucocytes was studied with a radioisotopic method . The following antibiotics were tested in concentrations obtained in the serum after standard administration: gentamicin, netilmicin, amikacin, oxacillin, clindamycin, vancomycin, teicoplanin, rifampicin, ansamycin, coumermycin, enoxacin, ciprofloxacin, two new fluoro-quinolones CI 934 and RO 236240, and LY 146032 a new lipopeptidic antibiotic . Among them, only coumermycin significantly decreased phagocytosis, this was dependent on concentration, strain and source of the polymorphonuclear leucocytes.

Eur J Haematol, 1987 Sep, 39(3), 259 - 66
A synthetic hemoregulatory peptide (HP5B) inhibits human myelopoietic colony formation (CFU-GM) but not leukocyte phagocytosis in vitro; Laerum OD et al.; A synthetic analog of a hemoregulatory peptide associated with mature human granulocyte (HP5B) has been investigated for inhibitory effects on human myelopoietic stem cells in vitro . In addition, it has been tested for effects on phagocytosis by human granulocytes and monocytes by use of an automatic flow cytometric method . A dose-dependent inhibition of colony formation was found after preincubation of bone marrow cells for 1 h at 37 degrees C in the range 10(7) -10(-11) mol/l . Above or below these concentrations, no inhibitory effects were seen . The degree of inhibition varied from experiment to experiment, indicating variable responsiveness of the donor cells . Maximal effect was of magnitude 90% inhibition, and the optimal dose was 10(7) mol/l . The peptide had no effect on the kinetics of phagocytosis by measurements of the uptake of fluorescent Zymosan particles or Staphylococcus aureus . This may indicate a selective effect on the precursor cells, with no effect on the functional state of their progeny, the granulocytes and monocytes.

Antimicrob Agents Chemother, 1987 Sep, 31(9), 1444 - 5
Single- and combination-antibiotic therapy for experimental endocarditis caused by methicillin-resistant Staphylococcus aureus; Rodriguez A et al.; The efficacy of fosfomycin in combination with vancomycin or gentamicin was evaluated in experimental endocarditis caused by methicillin-resistant Staphylococcus aureus . After 5 days of therapy, both combinations proved to be highly effective since all rabbits had sterile vegetations.

J Burn Care Rehabil, 1987 Sep-Oct, 8(5), 381 - 3
In vivo comparison of two silver sulfadiazine antimicrobial agents on burn wound infection; Velanovich V et al.; A controversy exists with regard to the relative efficacy of two preparations of silver sulfadiazine (AgSD), Silvadene and Flint's Silver Sulfadiazine Cream . We compared the susceptibility of Staphylococcus aureus and clinical Gram-positive cocci, Gram-negative rods, and mixed floral isolates by the Nathan's agar well diffusion method and found no differences . However, when S aureus-infected rat burn wounds were treated with these antimicrobial creams over a period of ten days, Silvadene significantly lowered bacterial counts, whereas results after treatment with Flint's Silver Sulfadiazine Cream were no different from those of the control group, which received no treatment . These data imply that Silvadene controls S aureus-generated burn wound infections better than the Flint product.

J Vet Pharmacol Ther, 1987 Sep, 10(3), 233 - 40
Florfenicol in non-lactating dairy cows: pharmacokinetics, binding to plasma proteins, and effects on phagocytosis by blood neutrophils; Bretzlaff KN et al.; Serial blood samples were collected and plasma concentrations of florfenicol (FLO) were measured following the administration of an intravenous bolus of 50 mg/kg FLO to five healthy non-lactating dairy cows . A triexponential equation provided the best fit of the data for four of the five cows . The mean value for beta corresponded to a half-life of 3.2 h . The mean apparent volume of distribution was 0.67 l/kg, and the mean body clearance was 0.15 l/kg/h . The extent of binding of FLO to bovine plasma proteins was determined in vitro at concentrations of 5 micrograms/ml and 50 micrograms/ml by equilibrium dialysis and ultrafiltration . The drug was 18% and 19% bound by equilibrium dialysis, and 23% and 19% bound by ultrafiltration, at 5 micrograms/ml and 50 micrograms/ml, respectively . Phagocytosis of 32phosphorus-labelled Staphylococcus aureus by bovine blood neutrophils was compared in vitro between neutrophils incubated in phosphate-buffered saline alone or in combination with 5, 125, or 1000 micrograms/ml chloramphenicol or FLO . There was no significant effect of chloramphenicol at any concentration . Florfenicol significantly inhibited phagocytosis at all concentrations, but the percentage inhibition was small . The clinical significance, if any, of this effect of FLO remains to be demonstrated.

J Trauma, 1987 Sep, 27(9), 1061 - 5
Resistance of rapidly expanded random skin flaps to bacterial invasion; Barker DE et al.; This study tested the hypothesis that rapidly expanded random pattern skin flaps demonstrate enhanced resistance to bacterial invasion from intradermal injection of Staphylococcus aureus in a porcine model . Sites for a 6 X 12 cm expanded, a 6 X 9 cm sham-expanded, and a 6 X 6 cm acute random pattern skin flap were outlined, but not elevated, on the backs of 14 white pigs . A 450-cc tissue expander inserted beneath the panniculus carnosus at the site for expansion was sequentially filled to the limits of skin viability each day for 5 days . At the sham site a tissue expander was similarly inserted but left unexpanded; the acute flap site was left undisturbed . On day 8, flaps were elevated, immediately sutured in place, and 0.1 ml of saline solution containing 10(7) Staphylococcus aureus inoculated intradermally at four corresponding sites in each flap and at three sites in normal skin . In seven animals these sites were at the proximal base of the flaps; in seven others the sites were distal . The resulting areas of erythema and skin ulceration were measured on each of the next 3 days and the measurements compared . At corresponding proximal sites, the mean area of erythema and ulceration measured over the next 3 days in expanded flaps (31.2 mm2) and in sham-expanded flaps (33.8 mm2) was significantly less than in acute flaps and control skin (47.5 mm2, 43.4 mm2, p less than 0.05) . Measurements in the rapidly expanded flaps were not significantly different than those in the sham-operated expanded flaps.(ABSTRACT TRUNCATED AT 250 WORDS)

J Trauma, 1987 Sep, 27(9), 1051 - 4
Influence of internal fixation on wound infections; Grewe SR et al.; With increasing frequency trauma surgeons are advocating early internal fixation in open fractures . The effect of the fixation devices on the infection rate in contaminated wounds remains a concern as our clinical experience in this area has been mixed . To study the effects of internal fixation on bone infections a 3.5-mm stainless steel screw was inserted into rabbit femurs and the wounds contaminated with Staphylococcus aureus . The controls had the screw hole drilled and taped but the screw was not inserted . Thirty of 49 rabbits receiving the screw subsequently became infected whereas 19 of 56 control animals developed an infection . The difference was significant at the 0.05 confidence level.

J Clin Microbiol, 1987 Sep, 25(9), 1587 - 90
Influence of lysozyme on aggregation of Staphylococcus aureus; Millar MR et al.; The conditions under which lysozyme aggregates Staphylococcus aureus were studied . Lysozyme was found to aggregate S . aureus at concentrations found in human tear secretions . Aggregate size depended upon lysozyme concentration, ionic strength, and bacterial concentration . There was a low level of adherence of S . aureus to corneal epithelial cells, and the adherence of a recent clinical isolate was not influenced by lysozyme concentrations found in human tear secretions . Lysozyme may enhance bacterial clearance from the corneal surface of the eye by promoting particle aggregation.

J Cell Biol, 1987 Sep, 105(3), 1241 - 51
Biogenesis of the rat hepatocyte plasma membrane in vivo: comparison of the pathways taken by apical and basolateral proteins using subcellular fractionation; Bartles JR et al.; We have used pulse-chase metabolic radiolabeling with L-{35S}methionine in conjunction with subcellular fractionation and specific protein immunoprecipitation techniques to compare the posttranslational transport pathways taken by endogenous domain-specific integral proteins of the rat hepatocyte plasma membrane in vivo . Our results suggest that both apical (HA 4, dipeptidylpeptidase IV, and aminopeptidase N) and basolateral (CE 9 and the asialoglycoprotein receptor {ASGP-R}) proteins reach the hepatocyte plasma membrane with similar kinetics . The mature molecular mass form of each of these proteins reaches its maximum specific radioactivity in a purified hepatocyte plasma membrane fraction after only 45 min of chase . However, at this time, the mature radiolabeled apical proteins are not associated with vesicles derived from the apical domain of the hepatocyte plasma membrane, but instead are associated with vesicles which, by several criteria, appear to be basolateral plasma membrane . These vesicles: (a) fractionate like basolateral plasma membrane in sucrose density gradients and in free-flow electrophoresis; (b) can be separated from the bulk of the likely organellar contaminants, including membranes derived from the late Golgi cisternae, transtubular network, and endosomes; (c) contain the proven basolateral constituents CE 9 and the ASGP-R, as judged by vesicle immunoadsorption using fixed Staphylococcus aureus cells and anti-ASGP-R antibodies; and (d) are oriented with their ectoplasmic surfaces facing outward, based on the results of vesicle immunoadsorption experiments using antibodies specific for the ectoplasmic domain of the ASGP-R . Only at times of chase greater than 45 min do significant amounts of the mature radiolabeled apical proteins arrive at the apical domain, and they do so at different rates . Approximate half-times for arrival are in the range of 90-120 min for aminopeptidase N and dipeptidylpeptidase IV whereas only 15-20% of the mature radiolabeled HA 4 associated with the hepatocyte plasma membrane fraction has become apical even after 150 min of chase . Our results suggest a mechanism for hepatocyte plasma membrane biogenesis in vivo in which all integral plasma membrane proteins are shipped first to the basolateral domain, followed by the specific retrieval and transport of apical proteins to the apical domain at distinct rates.

Am J Kidney Dis, 1987 Sep, 10(3), 241 - 9
Long-term survivors on peritoneal dialysis; Zimmerman SW et al.; To obtain information about predialysis characteristics and long-term outcome of patients on peritoneal dialysis (PD) for more than 4 years, we reviewed all patients starting PD who performed continuous ambulatory peritoneal dialysis (CAPD) and were at risk for more than 4 years . Sixty-two patients started; 42% were diabetic and 35% over age 60 . Three recovered renal function, seven received transplants, and 12 switched to hemodialysis . Nineteen survived more than 4 years (long-term survivors, LTS), eight diabetic and 12 male . Twenty-one died on CAPD in less than 4 years (short-term survivors, STS) . In comparison to STS, LTS were younger, with less prior cardiac disease, yet had higher predialysis serum creatinine values and lower hematocrits . LTS were observed for a mean of 65.3 +/- 3 months (48 to 91 months), and STS for a mean of 21 +/- 2 months . When compared to STS, LTS had fewer hospital days, hospital days for peritonitis, and a lower peritonitis rate, although the incidence of Staphylococcus aureus peritonitis was greater in LTS . Cardiovascular and thromboembolic events were less frequent in LTS, but bone fractures were seen more often in the LTS diabetic patients . Weight gain, especially in males, and hernias were noted in both groups . BP improved, and vision was maintained in both groups . Non-PD-related infections causing hospitalization were low in both groups . Improved mean hematocrit and hemoglobin A, values were seen only in LTS . Mean serum cholesterol values increased with time in LTS . This study reveals that potentially high-risk patients such as diabetics and the elderly can have prolonged survival on PD.(ABSTRACT TRUNCATED AT 250 WORDS)

J Surg Res, 1987 Sep, 43(3), 246 - 52
The effect of indomethacin on burn-induced immunosuppression; Latter DA et al.; Recent interest in the role of prostaglandin inhibitors as immunomodulators following major injury prompted us to study the effect of indomethacin on burn-induced immunosuppression in rats as measured by the delayed-type hypersensitivity (DTH) skin test response, ability to contain an intradermal bacterial challenge (10(8) Staphylococcus aureus 502A injected intradermally), and overall survival from spontaneous burn wound sepsis . Fifty male Sprague-Dawley rats sensitized to keyhole limpet hemocyanin (KLH) were subjected to a 30% full-thickness scald burn . Group 1 (n = 24) received indomethacin at 0.5 mg/kg intraperitoneally once daily with the first dose given immediately following the burn . Group 2 (n = 24) received vehicle only . Prostaglandin E2 measured by radioimmunoassay on day 17 was 2553 +/- 832 pcg/ml serum (+/- SEM) in the vehicle group and 1042 +/- 231 pcg/ml in the indomethacin group (P = 0.058, unpaired t test) . Burn injury induced a decrease in the DTH response to KLH and an increase in the Staph lesion size (P less than 0.05) which was not corrected by indomethacin treatment . All animals developed spontaneous burn wound sepsis by day 14 . Survival after 17 days in the indomethacin group was 100% compared to that of the vehicle group, 79%, P less than 0.05 (Fisher exact test) . We conclude that despite unmeasurable corrections of the burn-induced suppression of the DTH response and local nonspecific bacterial defenses, low-dose indomethacin improves survival following burn sepsis.

Invest Ophthalmol Vis Sci, 1987 Sep, 28(9), 1553 - 8
Corneal antibody levels to ribitol teichoic acid in rabbits immunized with staphylococcal antigens using various routes; Mondino BJ et al.; Although Staphylococcus aureus is an important cause of infectious diseases of the eye and hypersensitivity lesions of the cornea, little is known about ocular immunity to this pathogen . Using an enzyme-linked immunosorbent assay, we measured antibody titers to ribitol teichoic acid, the major antigenic determinant of S . aureus, in corneas as well as serum and tears after immunizing rabbits using the following routes: intradermal injection of cell wall mixed with complete Freund's adjuvant, subconjunctival injection of cell wall mixed with complete Freund's adjuvant, topical application of cell wall to the eye or topical application of viable S . aureus to the eye . IgG titers to ribitol teichoic acid were found consistently in corneas after intradermal and subconjunctival immunization with cell wall and topical immunization with viable S . aureus . After intradermal immunization with cell wall, IgG titers in cornea were higher than tears but lower than serum, which was presumably the source of the IgG antibodies for the cornea . After subconjunctival immunization with cell wall or topical immunization with viable S . aureus, IgG titers in corneas were higher than tears and generally higher than serum, suggesting that the ocular tissues were a local source of IgG . On the other hand, IgA titers to ribitol teichoic acid were found in tears but not in serum and were found only occasionally in corneas, suggesting that IgG responses to staphylococcal antigens may be more important than IgA responses in the cornea . The results of this study suggest that corneal antibodies to ribitol teichoic acid may be influenced by exposure to staphylococcal antigens not only in the external eye but also at sites remote from the eye.

Infect Immun, 1987 Sep, 55(9), 2191 - 7
Chemical characterization and immunogenicity of capsular polysaccharide isolated from mucoid Staphylococcus aureus; Lee JC et al.; In this study we report the isolation and purification of the capsular polysaccharide elaborated by Staphylococcus aureus SA1 mucoid . The capsule was isolated from bacterial extracts and culture supernatants by a series of ethanol precipitations and enzyme digestions, followed by ion-exchange chromatography . Teichoic acid contamination was eliminated by oxidation with sodium metaperiodate, and the final product eluted in the void volume of a Sephacryl S-300 column . The purified capsular polysaccharide was analyzed by gas-liquid chromatography-mass spectroscopy, 13C and 1H nuclear magnetic resonance, amino acid analysis, immunelectrophoresis, and numerous biochemical assays . The major constituents of the capsule were 2-acetamido-2-deoxy-alpha-galacturonic acid (4-O linked), 2-acetamido-2-deoxy-alpha-fucose (3-O linked), and taurine . The polysaccharide also contained O-acetyl groups which were removed by mild alkaline hydrolysis . Serologically and biochemically, the capsule from strain SA1 mucoid appeared very similar to that produced by strain M . Purified capsular polysaccharide was immunogenic in both rabbits and mice . The optimal immunizing dose in mice was 0.1 microgram of purified capsular polysaccharide administered intraperitoneally . SA1 mucoid resisted opsonophagocytic killing by human leukocytes and complement . However, antibodies raised to the purified capsular polysaccharide neutralized the antiphagocytic effect of the capsule.

Infect Immun, 1987 Sep, 55(9), 2155 - 63
Ingestion of Staphylococcus aureus by bovine endothelial cells results in time- and inoculum-dependent damage to endothelial cell monolayers; Vann JM et al.; Cultured endothelial cells phagocytize Staphylococcus aureus, but the resultant effects are unknown . Monolayers of cultured bovine endothelial cells with or without {3H}adenine label were exposed to 100, 10, or 1 S . aureus organism per endothelial cell for 3.5 h . Lysostaphin was then applied to all cultures to destroy extracellular but not phagocytized S . aureus . In cultures treated for only 20 min with lysostaphin, S . aureus multiplied exponentially after a 9- to 12-h lag period . In cultures treated continuously with lysostaphin, numbers of S . aureus remained constant or decreased . These results indicate that S . aureus became extracellular and multiplied but did not multiply intracellularly . In parallel experiments, the release of 3H-adenine from prelabeled endothelial cell monolayers was assayed to indicate cytotoxicity . Results indicated that the loss of 3H-adenine from endothelial cell monolayers depended on the following: (i) the size of the S . aureus inoculum, (ii) the strain of S . aureus, and (iii) the length of time after exposure to S . aureus . S . aureus endocarditis and persistent septicemia could arise, at least in part, from ingestion of S . aureus by host endothelium . The intracellular location would afford S . aureus protection from host defenses and antibiotics . Eventual damage to endothelial cells could expose collagen, thus resulting in platelet adherence and vegetation formation . Intracellular S . aureus would be continuously released into the circulation, possibly accounting for the persistent bacteremia that is found in S . aureus endovascular infections.

Antimicrob Agents Chemother, 1987 Sep, 31(9), 1426 - 8
Role of beta-lactamase in expression of resistance by methicillin-resistant Staphylococcus aureus; Boyce JM et al.; Of 27 unique clinical isolates of methicillin-resistant Staphylococcus aureus, only 4 were homogeneously resistant, and all 4 produced little or no beta-lactamase . Among heterogeneously resistant strains, those most resistant to beta-lactam antibiotics produced the most beta-lactamase . Similar genes may regulate production of the low-affinity penicillin-binding protein and beta-lactamase.

Antimicrob Agents Chemother, 1987 Sep, 31(9), 1423 - 5
Increased susceptibility to cephamycin-type antibiotics of methicillin-resistant Staphylococcus aureus defective in penicillin-binding protein 2; Murakami K et al.; A methicillin-resistant strain of Staphylococcus aureus which produced low-affinity penicillin-binding protein 2' (PBP 2') spontaneously lost PBP 2 when the strain was cultivated at 43 degrees C overnight . At 37 degrees C, the mutant had increased susceptibility to cephamycin-type beta-lactams, which showed high affinity for PBP 4 . This result suggests that inhibition of PBP 4, in addition to that of PBP 2, is necessary to kill methicillin-resistant strains.

Antimicrob Agents Chemother, 1987 Sep, 31(9), 1307 - 11
Production of low-affinity penicillin-binding protein by low- and high-resistance groups of methicillin-resistant Staphylococcus aureus; Murakami K et al.; Methicillin- and cephem-resistant Staphylococcus aureus (137 strains) for which the cefazolin MICs are at least 25 micrograms/ml could be classified into low-resistance (83% of strains) and high-resistance (the remaining 17%) groups by the MIC of flomoxef (6315-S), a 1-oxacephalosporin . The MICs were less than 6.3 micrograms/ml and more than 12.5 micrograms/ml in the low- and high-resistance groups, respectively . All strains produced penicillin-binding protein 2' (PBP 2'), which has been associated with methicillin resistance and which has very low affinity for beta-lactam antibiotics . Production of PBP 2' was regulated differently in low- and high-resistance strains . With penicillinase-producing strains of the low-resistance group, cefazolin, cefamandole, and cefmetazole induced PBP 2' production about 5-fold, while flomoxef induced production 2.4-fold or less . In contrast, penicillinase-negative variants of low-resistance strains produced PBP 2' constitutively in large amounts and induction did not occur . With high-resistance strains, flomoxef induced PBP 2' to an extent similar to that of cefazolin in both penicillinase-producing and -negative strains, except for one strain in which the induction did not occur . The amount of PBP 2' induced by beta-lactam antibiotics in penicillinase-producing strains of the low-resistance group correlated well with resistance to each antibiotic . Large amounts of PBP 2' in penicillinase-negative variants of the low-resistance group did not raise the MICs of beta-lactam compounds, although these strains were more resistant when challenged with flomoxef for 2 h . Different regulation of PBP 2' production was demonstrated in the high- and low-resistance groups, and factor(s) other than PBP 2' were suggested to be involved in the methicillin resistance of high-resistance strains.

Drug Intell Clin Pharm, 1987 Sep, 21(9), 728 - 32
Antibiotic prophylaxis in open-heart surgery patients: comparison of cefamandole and cefuroxime; Peterson CD et al.; The efficacy of cefamandole and cefuroxime in preventing postoperative wound infections was compared in 3037 patients undergoing open-heart surgery . Antibiotic prophylaxis in 1467 patients having coronary artery bypass and valve replacement surgery was cefamandole 2 g iv preoperatively followed by 2 g q6h for five days postoperatively; 1570 patients received cefuroxime 1.5 g iv preoperatively then 1.5 g iv q 12h for three days postoperatively . Postoperative wound infections (sternal and leg wounds) were studied in each treatment group . In the cefamandole study group, 27 patients (1.8 percent) developed postoperative wound infections (9 sternal and 18 leg wounds) . In the cefuroxime treatment group, 19 patients (1.2 percent) developed postoperative wound infections (9 sternal and 10 leg wounds) . Overall, no statistical difference was found between the two antibiotics in preventing postoperative wound infections . However, in patients having valve replacement surgery, cefuroxime was found statistically more effective than cefamandole prophylaxis in preventing sternal wound infections (no infections in 284 patients compared with five infections in 205 patients, respectively, p = 0.01) . The most common organism isolated from infected wounds with cefamandole was Staphylococcus aureus followed by S . epidermidis compared with cefuroxime which had S . epidermidis followed by S . aureus . Cefuroxime was found to be as effective as cefamandole and considerably less expensive in preventing postoperative wound infections in patients undergoing open-heart surgery.

J Gen Microbiol, 1987 Sep, 133 ( Pt 9), 2689 - 98
Production and immunological characterization of a monoclonal antibody to Trichophyton quinckeanum: interaction with phosphorylcholine-bearing components; Calderon RA et al.; A monoclonal antibody (Tq-1) that interacts with the phosphorylcholine (PC)-bearing antigens of Trichophyton quinckeanum was produced by fusion of myeloma cells (JKAg-8) with spleen cells of BALB/c mice immunized with an alum-precipitated fraction of T . quinckeanum cytoplasmic antigen . It was characterized as an IgM class antibody by immunodiffusion using anti-Ig heavy chain specific reagents, ELISA using immunoglobulin-specific peroxidase-conjugated antibodies, and by gel filtration chromatography; it showed high affinity for Staphylococcus aureus protein-A . Interaction of Tq-1 with PC-like antigens of T . quinckeanum was demonstrated by inhibition studies using ELISA, immunoblotting, immunoprecipitation and immuno electron microscopy techniques . The binding activity of Tq-1 antibody with a range of dermatophyte proteins was completely inhibited by prior incubation with PC hapten . Moreover, dermatophyte antigens reacting with the monoclonal antibody reacted strongly with sera from chronically infected mice . Dermatophyte antigens derived from both young (24 h) and old (20 d) cultures reacted with Tq-1 and this binding was inhibited by PC, suggesting that Tq-1 target antigen PC appears at an early stage during fungal growth and remains throughout its life.

Zh Mikrobiol Epidemiol Immunobiol, 1987 Sep, (9), 61 - 4
{Antibodies to Staphylococcus aureus teichoic acids in the pathogenesis of chronic osteomyelitis}; Urazgil'deev ZI et al.; The titers of antibodies to S . aureus cell-wall teichoic acids have been determined in 97 orthopedic and traumatic patients with purulent diseases, differing by the activity of the process, by means of the enzyme immunoassay . These antibodies appeared in the patients' blood in active osteomyelitic process of staphylococcal etiology.

Rev Infect Dis, 1987 Sep-Oct, 9 Suppl 5, S482 - 9
On the pathogenesis of toxic shock syndrome; Kass EH et al.; Understanding of the pathogenesis of toxic shock syndrome (TSS) has come from the juxtaposition of epidemiologic, clinical, immunologic, and physiologic studies . A hypothesis has been developed for the pathogenesis of menstrually related TSS . Certain tampon fibers that are highly absorbent for water are also ion exchangers for magnesium ions . The latter ions uniquely affect the production of TSS toxin 1 (TSST-1) by appropriate strains of Staphylococcus aureus, with a marked increase in the amount of toxin when magnesium concentrations are limiting and suppression of toxin production when magnesium is in excess . Many epidemiologic features of TSS could be explained by this hypothesis . The absorbability of highly absorptive fibers is not affected by the addition of small amounts of magnesium sufficient to suppress production of TSST-1; thus absorption is distinguishable from toxin production in vitro . TSST-1 stimulates production of interleukin 1 and of tumor necrosis factor and is highly toxic when absorbed slowly . Like TSST-1, staphylococcal enterotoxins are lethal to rabbits when given by slow injection, and some enterotoxins may be more lethal than TSST-1.

Rev Infect Dis, 1987 Sep-Oct, 9(5), 891 - 907
Prospective study of 114 consecutive episodes of Staphylococcus aureus bacteremia; Mylotte JM et al.; From 1 April 1983 to 31 October 1985, 114 episodes of Staphylococcus aureus bacteremia (SAB) were identified in 111 patients at the Buffalo Veterans Administration Medical Center . Only 14% of the episodes were community-acquired, and 29% were due to methicillin-resistant strains . The commonest foci of SAB were intravascular catheters (33%), postoperative wounds (11%), skin infections (7%), and pulmonary infections (7%) . Complications were infrequent, with endocarditis in two patients and metastatic infection in one . Mortality due to SAB was 32%, with no difference in mortality between community-acquired and hospital-acquired SAB . Although not statistically significant, there was a trend of higher mortality for methicillin-resistant SAB (42%) than for methicillin-sensitive SAB (28%) and for patients with no focus of SAB (43%) than for those with a defined primary focus (28%) . A review of studies of SAB published since 1940 revealed several trends . SAB is now predominately a nosocomial infection; intravascular-catheter infection has become the commonest cause of SAB; with several exceptions, the risk of endocarditis in patients with SAB is low (5%-20%); mortality due to SAB has decreased over the past 40 years but not over the past 10 years.

Appl Environ Microbiol, 1987 Sep, 53(9), 2060 - 5
Survival and transport of bacteria in egg washwater; Pearson J et al.; An evaluation of methods for monitoring the quality of water used to wash eggs at grading stations was undertaken to improve maintenance of bacterial viability during overnight sample transport . Bacterial content of samples at analysis would then better reflect conditions at the time eggs were washed . The interactive effects of temperature and the highly alkaline water conditions upon viability were the subjects of this study . Nine transport methods were examined for their efficacy in recovering total and coliform bacteria from recycled water used to wash eggs, and these were compared with samples analyzed at two commercial egg grading stations . Samples were shipped under test to the laboratory for analysis the following day . The survival of Staphylococcus aureus and Escherichia coli was also examined, but in a synthetic washwater matrix under various combinations of temperature (6 to 32 degrees C) and pH (9.5 to 10.5) to determine whether there was likely to be a different response to variations in transport treatment among gram-positive and -negative bacteria . S . aureus was much more resistant to the lethal effects of high pH and moderate temperature than E . coli . These results indicated that samples of high pH should be held (transported) at less than or equal to 13 degrees C to optimize bacterial survival . Considering cost, ease of manipulation, and the ability to protect both coliforms and the bacterial population as a whole, the method of choice for transport of industrial samples was the direct addition of washwater to containers in which powdered KH2PO4 and Na2S2O3 had been placed to yield final concentrations, when dissolved, of 0.2 and 0.05% (wt/vol), respectively.

Pathol Biol (Paris), 1987 Sep, 35(7), 1057 - 61
{Comparison of minimum inhibitory concentrations of group M penicillins against Staphylococcus aureus . Practical impact on diagnosis and treatment}; de Rautlin de la Roy Y et al.; The MICs were determined in usual conditions and in special conditions for the detection of heterogeneous methicillin resistance . The study involved 40 strains of Staphylococcus aureus of the 3 categories of homogeneous resistant, heterogeneous resistant and sensitive strains . The MICs of the 4 main compounds of the group M penicillins, methicillin, oxacillin, cloxacillin and dicloxacillin, were lower in the presence of a higher number of chlorine atoms on the molecule . The heterogeneous strains were even sensitive to dicloxacillin . The MIC for a notable fraction of resistant strains were lower than the critical value, i.e . these strains entered the category of dicloxacillin sensitive even in reading conditions theoretically leading to the detection of heterogeneous resistance . These in vitro observations are of value in the presence of methicillin sensitive strains, since antibiotics containing one or two chlorine atoms will probably be more therapeutically effective . But the bacteriologist should pay more careful attention to the detection of methicillin resistant and heteroresistant strains, since the resistance in vivo is extended to other penicillinase-resistant penicillins or penicillins M.

Acta Paediatr Scand, 1987 Sep, 76(5), 769 - 74
Plasmid profiles of multi-resistant Staphylococcus aureus at a children's hospital; Tang J et al.; Twenty-six strains of multi-resistant Staphylococcus aureus (MRSA) recovered from November 1985 to February 1986 were screened for their plasmid profiles . Among them, 11 strains were recovered from 8 patients with definite nosocomial infections . The results showed that one MRSA strain carrying 5 plasmids with molecular weight ranging from 1.75 X 10(6) to 5 X 10(6) daltons was epidemic in our hospital while 3 other distinct strains caused cross infections in both the burn ward and the medical wards . The epidemic strain and two other hospital-acquired strains were found in identical form in the community, suggesting that the pattern of nosocomial MRSA can be affected by strains outside hospital . The plasmid profile analysis was employed to differentiate the epidemic isolates from the coisolates and to study the relation between clinical isolates . The method is apparently superior to antibiogram.

Surgery, 1987 Sep, 102(3), 477 - 84
Neutrophil activation after burn injury: contributions of the classic complement pathway and of endotoxin; Davis CF et al.; We attempt to elucidate the mechanisms of neutrophil (PMN) activation after burn injury . We previously reported prolonged elevations of PMN cell surface complement (C) opsonin receptor levels after burn trauma with a corresponding period of depressed PMN chemotaxis to C5a, which suggests that the C product, C5a, was responsible for PMN activation . However, a lack of direct correlation of C activation with C receptor levels soon after injury raised the possibility of a second PMN-activating substance . We therefore investigated the effect of endotoxin (LPS) on the expression of the C receptors (CR1 and CR3) by normal human PMNs . Concentrations from 0 to 50 ng/ml of LPS 026:B6 caused a dose response increase in the PMN surface expression of CR1 and CR3 as assessed by monoclonal antibody binding and indirect immunofluorescence . The relative CR1-dependent fluorescence rose from a mean of 50 to 385 and CR3 from 50 to 300 . Chelation by ethylenediaminetetra acetic acid (EDTA) did not influence this dose response, thus ruling out the possibility of C activation by LPS--an inference supported by the lack of complement activation observed with these concentrations of LPS in normal serum . A similar dose response was obtained in the absence of other cell types or serum, which implies a direct effect that mimicked that of C5a . To determine the mechanism of the later, prolonged C activation after burn injury, we next examined C activation products in 22 patients with burn injuries . Elevations of plasma C3a desArg were present and persisted for 50 days . Elevations were at maximum levels on days 9 through 13 postburn (mean +/- standard error of mean {SEM}, 496 +/- 47 ng/ml versus normal 113 +/- 32; p less than 0.01) . These were accompanied by elevations of C4a desArg (917 +/- 154 ng/ml versus normal 424 +/- 50; p less than 0.01), which are indicative of classic pathway activation . Finally, we examined PMN function, phagocytosis and percentage killing of Staphylococcus aureus, and found PMN function to be unaltered in the 22 patients . Thus PMN activation after burn injury appears to be caused by LPS soon after injury and by C5a later after injury and affects only selected PMN functions.

Proc Natl Acad Sci U S A, 1987 Sep, 84(18), 6404 - 7
Glucose transport and antilipolysis are differentially regulated by the polar head group of an insulin-sensitive glycophospholipid; Kelly KL et al.; A glycophospholipid has been purified from rat liver membranes, which copurified with an insulin-sensitive glycophospholipid isolated from H35 hepatoma cells . The polar head group of this glycophospholipid, which is a phosphooligosaccharide, was generated by treatment with a phosphatidylinositol-specific phospholipase C from Staphylococcus aureus . There was an "insulin-like" inhibitory effect of this phosphooligosaccharide on isoproterenol-stimulated lipolysis in adipocytes, whereas there was no effect on glucose oxidation under conditions that measure glucose transport . The antilipolytic effect of this phosphooligosaccharide was demonstrated in intact adipocytes . There was a linear correlation between the concentration of phosphooligosaccharide and its antilipolytic effect, the magnitude and time course of which were similar to that obtained with physiological concentrations of insulin . Submaximal concentrations of insulin and phosphooligosaccharide produced an additive antilipolytic effect . The antilipolytic effect of the phosphooligosaccharide was demonstrated only after release of this compound from the precursor glycophospholipid with phosphatidylinositol-specific phospholipase C, and the activity of the phosphooligosaccharide was sensitive to alkali . It is proposed that this phosphooligosaccharide plays a role in mediating certain insulin actions.

J Neurosurg, 1987 Sep, 67(3), 438 - 45
Bacterial adhesion to cerebrospinal fluid shunts; Guevara JA et al.; Bacterial adherence to cerebrospinal fluid (CSF) shunts was analyzed in vivo and in vitro . Scanning electron micrographs (SEM's) of catheters removed from pediatric patients with shunts infected by Staphylococcus aureus or Klebsiella pneumoniae revealed numerous bacterial cells and microcolonies, leukocytes, and erythrocytes attached to the CSF catheters' inner walls, as well as the existence of surface irregularities, such as fissures, rugosities, and holes . Permeability analyses and SEM's demonstrated that catheters develop physical alterations over the period of implantation . Different bacterial strains presented a different in vitro adherence to CSF shunts, suggesting that this attachment may be affected by specific properties of the outer structures of each strain . The attachment of microbial pathogens to CSF shunts seems to contribute to the persistence of bacterial cells within a catheter and the onset of recurrent shunt infection . This study demonstrated that some bacteria can remain attached within shunts in vitro despite a CSF flow at rates up to 200 times higher than those normally demonstrated in vivo . Furthermore, surface irregularities found throughout this study may help to anchor and hide bacterial microcolonies . Based on these findings, it seems advisable to remove an infected shunt and to replace it with a new one after proper antimicrobial therapy, in order to prevent recurrent infections.

Ann Inst Pasteur Microbiol, 1987 Sep-Oct, 138(5), 561 - 7
Isolation of type 5 capsular polysaccharide from Staphylococcus aureus; Fournier JM et al.; Clinical surveys have indicated the predominance of type 5 and type 8 capsular polysaccharides among clinical isolates of Staphylococcus aureus . The type 5 capsular polysaccharide was extracted from three clinical isolates of S . aureus grown on solid media, and purified by ion exchange chromatography and gel filtration . Chemical analysis showed that the type 5 capsular polysaccharide is composed of N-acetylfucosamine and N-acetylhexosaminuronic acid . Immunodiffusion and 13C nuclear magnetic resonance experiments showed that the type 5 capsular polysaccharide is both immunologically and chemically distinct from the type 8 capsular polysaccharide and the teichoic acid of S . aureus.

Biochem J, 1987 Sep 1, 246(2), 431 - 9
Identification and purification from bovine brain of a guanine-nucleotide-binding protein distinct from Gs, Gi and Go; Waldo GL et al.; A guanine-nucleotide-binding protein (G-protein) was purified from cholate extracts of bovine brain membranes by sequential DEAE-Sephacel, Ultrogel AcA-34, heptylamine-Sepharose and Sephadex G-150 chromatography . Guanosine 5'-{gamma-{35S}thio}triphosphate (GTP{35S})-binding activity copurified with a 25,000 Da peptide and a 35,000-36,000 Da protein doublet . Neither pertussis toxin nor cholera toxin catalysed the ADP-ribosylation of a protein associated with the GTP{35S}-binding activity . Photoaffinity labelling of the purified protein with 8-azido{gamma-32P}GTP indicated that the GTP-binding site resides on the 25,000 Da protein . The 35,000-36,000 Da protein doublet was electrophoretically indistinguishable from the beta-subunits of other GTP-binding proteins, and the 36,000 Da protein was recognized by antiserum to oligomeric Gt . The purified protein specifically bound 17.2 nmol of GTP{35S}/mg of protein . The Kd of the binding site for radioligand was approx . 15 nM . The brain GTP-binding protein co-migrated during SDS/polyacrylamide-gel electrophoresis with a GTP-binding protein, named Gp, purified from human placenta {Evans, Brown, Fraser & Northup (1986) J . Biol . Chem . 261, 7052-7059}, and cross-reacted with antiserum raised against the placental protein, but not with antiserum raised to brain Go . SDS/polyacrylamide-gel electrophoresis of the brain and placental GTP-binding proteins in the presence of Staphylococcus aureus V8 protease yielded identical peptide maps.

Immunobiology, 1987 Sep, 175(3), 172 - 82
B cell maturation in chronic lymphocytic leukemia . II . Clonal heterogeneity in the response to various B cell activators and recombinant cytokines (rIFN-gamma, rIFN-alpha, rIL2); Pfeffer K et al.; Maturation of B cells into immunoglobulin (Ig)-secreting cells is influenced by antigen-nonspecific growth and differentiation factors . In the present study, we have analyzed the effect of several recombinant cytokines (rIL2, rIFN-alpha, rIFN-gamma) on the terminal differentiation of B cells from 10 patients with chronic lymphocytic leukemia (CLL) stimulated with three different B cell activators (phorbolester TPA, staphylococcus aureus Cowan I bacteria, or pokeweed mitogen) . Secretion of IgM, as determined by heavy-chain specific ELISA, varied widely among different CLL cell populations, but was induced in all cases by one or several of these activators . Importantly, this response was enhanced 2-10-fold in all patients by at least one of the tested recombinant cytokines alone, or a combination of rIL2 with rIFN-alpha or rIFN-gamma . The results illustrate the clonal heterogeneity of CLL B cell maturation in vitro and demonstrate that rIL2, rIFN-alpha or rIFN-gamma can act as a B cell differentiation factor, depending on the responsiveness (stage of differentiation) of the individual clonal B cell population.

Clin Exp Immunol, 1987 Sep, 69(3), 639 - 46
1,25-Dihydroxyvitamin D3-mediated inhibition of human B cell differentiation; Chen WC et al.; We have examined the mechanisms of 1,25-dihydroxyvitamin D3 (D3)-mediated inhibition of human B cell differentiation to immunoglobulin (Ig) secreting cells . B lymphocytes were purified from human tonsils and peripheral blood mononuclear cells . Mononuclear cells were separated into adherent cells and nonadherent cells . Cells were stimulated with Staphylococcus aureus Cowen I (SAC) or pokeweed mitogen (PWM) for 7 days and immunoglobulin production was measured by ELISA assay, 1,25-dihydroxyvitamin D3 was added at different times during cultures . 1,25-Dihydroxyvitamin D3, in a dose-dependent manner, inhibited both SAC and PWM-induced Ig production by mononuclear cells . The maximum inhibition was observed when 1,25-dihydroxyvitamin D3 was added at the beginning of culture, but inhibition could still be observed when 1,25-dihydroxyvitamin D3 was added on day 4 of cultures . The inhibitory effect of 1,25-dihydroxyvitamin D3 on Ig production was significantly greater on mononuclear cells than on nonadherent cells . Addition of in vitro purified IL-1 to nonadherent cells enhanced 1,25-dihydroxyvitamin D3-induced inhibition of Ig production . 1,25-Dihydroxyvitamin D3 also inhibited the expression of IL-2 receptors on B cells activated with SAC . 1,25-dihydroxyvitamin D3 did not inhibit Ig production by SAC preactivated B cell blasts in response to T cell supernatants . These data suggest that vitamin D3 inhibits Ig production by inhibiting IL-2 receptor expression on B cells and via its effect on adherent macrophages . Vitamin D3 does not influence the effect of differentiation factors on activated B cells that have already expressed growth/differentiation factor receptors.

J Exp Med, 1987 Sep 1, 166(3), 786 - 91
Effect of tumor necrosis factor alpha on mitogen-activated human B cells; Kehrl JH et al.; In this study we demonstrate that the monocyte/macrophage product, tumor necrosis factor alpha (TNF-alpha), has significant in vitro effects of B cell function . It costimulated with anti-mu in the induction of B cell DNA synthesis, and it prolonged the DNA synthesis initiated in B cell cultures stimulated with the human B cell mitogen, Staphylococcus aureus Cowan strain I (SAC) . The addition of either IL-1 or IFN-gamma to TNF-alpha resulted in a substantial further increase in DNA synthesis . The addition of TNF-alpha to IL-2, a known inducer of SAC-activated B cell Ig secretion, resulted in a twofold enhancement in the amount of IL-2 stimulated B cell Ig secretion . Receptor binding studies with 125I-TNF-alpha demonstrate a marked increase in TNF-alpha binding sites after B cell activation (approximately 6,000 sites per cell, with an apparent Kd of 2.0 X 10(-10) M) . Thus, TNF-alpha may be an important factor in human B cell function and is likely to interact with other T cell and monocyte derived cytokines in the regulation of human B cell proliferation and Ig production.

J Bacteriol, 1987 Sep, 169(9), 3910 - 5
Nucleotide sequence of the epidermolytic toxin A gene of Staphylococcus aureus; O'Toole PW et al.; The nucleotide sequence of the eta gene, which codes for the epidermolytic toxin serotype A of Staphylococcus aureus TC16, is reported . The coding sequence of 840 nucleotides specifies a protein which, when secreted, has a predicted molecular weight of 26,950 . The sequence of eta and the deduced amino acid sequence of the toxin have been compared with those of epidermolytic toxin serotype B . The coding sequences have 52% identical residues, and the polypeptides have 40% identical residues . Amino acid residues have been conserved in the areas of the proteins which correspond to major hydrophobic domains, whereas the regions likely to specify antigenic determinants occur in hydrophilic sequences that have diverged . The level of expression of epidermolytic toxin A in S . aureus 8325-4 was shown to be dependent on the integrity of a regulatory gene called agr.

J Bacteriol, 1987 Sep, 169(9), 3904 - 9
Sequence determination and comparison of the exfoliative toxin A and toxin B genes from Staphylococcus aureus; Lee CY et al.; The DNA encoding the exfoliative toxin A gene (eta) of Staphylococcus aureus was cloned into bacteriophage lambda gt11 and subsequently into plasmid pLI50 on a 1,391-base-pair DNA fragment of the chromosome . Exfoliative toxin A is expressed in the Escherichia coli genetic background, is similar in length to the toxin purified from culture medium, and is biologically active in an animal assay . The nucleotide sequence of the DNA fragment containing the gene was determined . The protein deduced from the nucleotide sequence is a polypeptide of 280 amino acids . The mature protein is 242 amino acids . The DNA sequence of the exfoliative toxin B gene was also determined . Corrections indicate that the amino acid sequence of exfoliative toxin B is in accord with chemical sequence data.

J Vasc Surg, 1987 Sep, 6(3), 235 - 9
Primary graft infections; Edwards WH Jr et al.; An amputation rate of 8% to 52% and a mortality rate of 13% to 58% make vascular prosthetic graft infections the most dreaded complication facing a vascular surgeon . In 1978 a randomized prospective double-blind study reported a statistically significant decrease in wound infections in patients treated with prophylactic antibiotics whereas the graft infection difference only approached statistical significance . The present study reviews 2614 arterial prosthetic grafts implanted from January 1975 through June 1986 . Twenty-four patients were identified as having a prosthetic graft infection, yielding an overall infection rate of 0.92% . Staphylococcus aureus was the most common organism, occurring in one third of the cases . The most common graft material was polytetrafluoroethylene (PTFE) (33%) followed by Dacron (29%), composite PTFE and Dacron (20%), and umbilical vein grafts (9%) . Diabetes was a common factor in one third of the patients . Symptoms of infection were present in 15 patients (63%) within 3 months of operation, with 11 patients showing symptoms within 30 days . The longest interval between operation and onset of symptoms was 48 months . Prophylactic antibiotics were administered to 22 of the 24 patients, but in only 7 of the 22 (29.5%) were they given according to our usual practice . All patients required removal of the infected prosthesis, with limb loss in 17% and death in 17%.

J Hosp Infect, 1987 Sep, 10(2), 165 - 72
A placebo-controlled trial of the effect of two preoperative baths or showers with chlorhexidine detergent on postoperative wound infection rates; Hayek LJ et al.; The effect of preoperative whole-body washing with chlorhexidine detergent on the incidence of postoperative wound infection was assessed in a placebo-controlled trial of 1989 patients . Patients bathed or showered with chlorhexidine, placebo, or conventional bar soap, on two occasions in the 24 h before operation . The overall infection rate for patients treated with chlorhexidine was 9%, against 12.8% in the bar soap and 11.7% in the placebo groups; in the 'clean' surgery group infections were 7.2% against 10.2% and 10%, respectively . The Staphylococcus aureus infection rate in the 'clean' group was 3% for chlorhexidine against 6% for bar soap.

J Hosp Infect, 1987 Sep, 10(2), 120 - 8
An outbreak of infection with a methicillin-resistant Staphylococcus aureus in a special care baby unit: value of topical mupirocin and of traditional methods of infection control; Davies EA et al.; This report deals with the problems associated with a high incidence rate of methicillin-resistant Staphylococcus aureus in low birth weight infants in a regional special care baby unit . Strict isolation facilities were not used and the outbreak was promptly brought under control by the use of intensive traditional methods of infection control and the use of topical mupirocin in a paraffin base.

Cell Immunol, 1987 Sep, 108(2), 343 - 55
Functional and biochemical characterization of B-cell differentiation factor (BCDF) produced by an HTLV-I-transformed human T-cell clone and demonstration of specific binding of the factor to a BCDF responsive cell line; Goldstein H et al.; We have previously described YA2, a human T-cell clone that secretes B-cell differentiation factor (BCDF) but not B-cell growth factor (BCGF) . The BCDFs secreted by YA2 and HTLV-I-transformed YA2 (TYA2) were functionally similar in their ability to stimulate Ig secretion by Staphylococcus aureus Cowan strain I-activated B cells and IgM secretion by SKW6.4 cells . In addition, they were biochemically similar with a MW of 30 kDa by high-performance liquid chromatography (HPLC) sieving, and a pI of 6.0-6.8 by isoelectric focusing . The BCDF activity was not blocked by antibodies to interleukin 2 and BCGF . BCDF was purified from TYA2 supernatant by sequential media protein immunoadsorption, flat bed isoelectric focusing, HPLC TSK 2000 sieving, and repeated immunoadsorption and was then iodinated . The iodinated material had functional BCDF activity and migrated to a single band at MW 30 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and at pI of 6.8 by polyacrylamide gel isoelectric focusing . 125I-BCDF purified in this manner bound specifically to a BCDF-responsive cell line and not to phytohemagglutinin-activated T cells . 125I binding to the BCDF-responsive cell line was competitively inhibitable by the addition of cold BCDF . Thus we have purified and characterized a factor with BCDF activity and demonstrated that this factor binds specifically to a BCDF-responsive cell line.

J Med Microbiol, 1987 Sep, 24(2), 139 - 44
Chromosome- and plasmid-mediated gentamicin resistance in Staphylococcus aureus encoded by Tn4001; Gillespie MT et al.; DNA sequences corresponding to the 4.7-kb gentamicin, tobramycin and kanamycin resistance (GmrTmrKmr) transposon Tn4001 have been detected on a series of nine structurally-related plasmids that mediate this phenotype in Australian isolates of Staphylococcus aureus . Tn4001 sequences have also been demonstrated on the chromosomes of GmrTmrKmr isolates that do not possess these plasmids, and the exhibited diversity of chromosomal sites occupied by this element implies that Tn4001 has transposed to the chromosome on numerous occasions in vivo . These results suggest that the rapid emergence of nosocomial GmrTmrKmr S . aureus in the early 1980s may have been the result of the transposition of Tn4001 from a chromosomal site to a readily disseminated plasmid.

FEBS Lett, 1987 Aug 31, 221(1), 167 - 71
Evolution of an inducible penicillin-target protein in methicillin-resistant Staphylococcus aureus by gene fusion; Song MD et al.; A new beta-lactam-inducible penicillin-binding protein (PBP) that has extremely low affinity to penicillin and most other beta-lactam antibiotics has been widely found in highly beta-lactam(methicillin)-resistant Staphylococcus aureus (MRSA) . The gene for this protein was sequenced and the nucleotide sequence in its promoter and close upstream area was found to show close similarity with that of staphylococcal penicillinase, while the amino acid sequence over a wide range of the molecule was found to be similar to those of two PBPs of Escherichia coli, the shape-determining protein (PBP 2) and septum-forming one (PBP 3) . Probably the MRSA PBP (Mr 76462) evolved by recombination of two genes: an inducible type I penicillinase gene and a PBP gene of a bacterium, causing the formation of a beta-lactam-inducible MRSA PBP.

Biochemistry, 1987 Aug 25, 26(17), 5239 - 44
Expression of human insulin-like growth factor I in bacteria: use of optimized gene fusion vectors to facilitate protein purification; Moks T et al.; Several fusions between the gene for human insulin-like growth factor I (IGF-I) and the genes for different IgG-binding fragments of staphylococcal protein A were assembled and compared regarding expression, secretion, and purification of the peptide hormone . After IgG affinity purification of the fusion proteins from the growth medium of Staphylococcus aureus or Escherichia coli, native IGF-I was released by cleavage of an Asn-Gly peptide bond with hydroxylamine . An optimized expression system based on a modified synthetic IgG-binding domain (z), resistant to hydroxylamine, gave the highest yield of fusion protein . After cleavage, the hormone could be separated from the IgG-binding moiety and from noncleaved fusion protein by a second passage through the IgG affinity column . The biological activity and the purity of the IGF-I obtained were confirmed by a radioreceptor assay, N-terminal sequence analysis, polyacrylamide gel electrophoresis, isoelectric focusing, and high-performance liquid chromatography.

Eur J Biochem, 1987 Aug 17, 167(1), 149 - 54
Improved purification of pyruvate decarboxylase from wheat germ . Its partial characterisation and comparison with the yeast enzyme; Zehender H et al.; An improved procedure was developed for the isolation of pyruvate decarboxylase from wheat germ . Its final step, an electrophoresis of the native apoenzyme in concave pore gradient polyacrylamide gels, followed by superficial activity-staining, produced two bands of different molecular masses and chain compositions . The high-molecular-mass band occurred in low quantity and consisted of, probably eight, apparently identical chains of Mr = 33,000, as judged from sodium dodecyl sulfate electrophoreses . The low-molecular-mass band contained two types of chains with Mr alpha = 63,000-65,000 and Mr beta = 61,000-62,000 . The N termini of both chains were threonine, whereas their C-terminal sequences were different: alpha, -(Val)-(Ser)-(Ala)-Leu; beta, -(His)-(Asp)-(Ala)-Ser . Their amino acid composition was too different to be compatible with our original concept of one chain being produced from the other by proteolytic shortening . Limited proteolysis by Staphylococcus aureus V8 proteinase yielded peptides partly identical size and partly quite different . In all properties investigated, the low-molecular-mass enzyme largely resembled yeast pyruvate decarboxylase; the holoenzyme appeared to possess (alpha beta)2 structure, the apoenzyme alpha beta . SH reagents inactivated the enzyme . Binding and fluorescence of 2-p-toluidinonaphthalene-6-sulfonate indicated a similar lipophilicity of the active site as found earlier for the yeast enzyme . 2-Hydroxy-5-nitrobenzyl modification of exposed tryptophan residues left the holoenzyme intact, but in the apoenzyme it destroyed most of the cofactor-binding ability and hence the activity . The strength of cofactor binding and the maximal specific activity were found somewhat lower than in yeast pyruvate decarboxylase.

Biochem J, 1987 Aug 15, 246(1), 121 - 9
Identification and partial characterization of two major proteins of Mr 47,000 synthesized by bovine retinal endothelial cells in culture; Canfield AE et al.; Biosynthetic experiments with cultured bovine retinal endothelial cells have identified a glycoprotein of Mr 47,000 (Gp47) as a major component secreted into the medium . Gp47 is a non-collagenous glycoprotein with a pI of 4.6-5.5, which does not bind to either gelatin-Sepharose or heparin-Sepharose but is retained by concanavalin A-Sepharose . The Mr of this species decreases to approx . 42,000 in the presence of tunicamycin, indicating that it contains asparagine-linked oligosaccharides . A second protein of Mr 47,000 (P47) is present in the cell layer/matrix of these cultured cells . The electrophoretic mobility of P47 remains unaltered when synthesized in the presence of tunicamycin . Peptide-mapping experiments using N-chlorosuccinimide and Staphylococcus aureus V8 proteinase demonstrate that Gp47 and P47 are distinct proteins, and are not related to colligin, a membrane-bound collagen-receptor protein of similar size, or to SPARC, a major secreted product of parietal endodermal cells and sparse cultures of aortic endothelial cells.

J Am Vet Med Assoc, 1987 Aug 15, 191(4), 425 - 30
Assessment of neutrophil function in dogs with primary ciliary dyskinesia; Morrison WB et al.; Compared with neutrophils from healthy dogs, neutrophils from 2 dogs with primary ciliary dyskinesia had increased distance of random migration, but fewer of the neutrophils migrated . The affected dogs had an increase in the numbers of Staphylococcus aureus phagocytized . Lymphocyte blastogenesis in the affected dogs in response to standard mitogens was considered to be normal.

J Immunol, 1987 Aug 15, 139(4), 1120 - 6
Interleukin 1 release by human monocytes treated with liposome-encapsulated lipopolysaccharide; Bakouche O et al.; Liposome therapy offers a unique method of delivering antifungals, antiprotozoans, and macrophage-activating factors directly to reticuloendothelial cells . Although liposomes are entering clinical trials, their effect on Il 1 synthesis and release has yet to be determined . The beneficial, as well as possible pathological, effects of liposome therapy may be due to IL 1 released by reticuloendothelial cells . This study examined the effects of multilamellar phospholipid vesicles on the synthesis and release of IL 1 from human peripheral blood monocytes . Liposomes consisting of phosphatidylcholine and phosphatidylserine in molar ratio of 7:3 did not stimulate IL 1 release, and did not diminish the level of IL 1 release when monocytes were subsequently stimulated with LPS or Staphylococcus aureus . Surprisingly, LPS encapsulated in liposomes failed to stimulate IL 1 release from monocytes . This defect was limited to the release of IL 1, because monocytes stimulated with liposomes containing LPS had control levels of intracellular (cytosolic) and membrane IL 1 . By contrast, LPS associated with lyophilized liposomes induced the secretion of IL 1 and behaved similarly to free LPS . These findings indicate that LPS activation of monocytes may involve not only the synthesis of IL 1 but activation of the secretory mechanism for IL 1, and that these two events could be independent, that the density of LPS molecules at the surface of the liposome may influence the degree of monocyte activation as measured by intracellular IL 1 synthesis, membrane accumulation, and secretion into the medium, and that intracellular lipids may not interfere with the secretory mechanism, because liposomes made with phospholipids differing in acyl chain length or the headgroup behaved identically.

J Biol Chem, 1987 Aug 15, 262(23), 11097 - 103
Human apolipoprotein B-100 heparin-binding sites; Weisgraber KH et al.; Seven distinct heparin-binding sites have been demonstrated on human apolipoprotein (apo) B-100 by using a combination of digestion with cyanogen bromide or Staphylococcus aureus V-8 protease and heparin-Sepharose affinity chromatography . Based on fragment analysis, the approximate boundaries of the seven binding sites are as follows: site A, residues 5-99; site B, residues 205-279; site C, residues 875-932; site D, residues 2016-2151; site E, residues 3134-3209; site F, 3356-3489; and site G, residues 3659-3719 . In sites E and F, two short regions enriched in basic amino acids have been identified, and it is likely that they are responsible for a major portion of the heparin-binding properties of these sites . The relative binding affinity of each of the seven sites was estimated in two ways . First, the affinity was assessed in a ligand blot assay using a 125I-labeled high-reactive heparin subfraction . Second, apoB-100 fragments generated by cyanogen bromide or S . aureus V-8 protease were separated into low- and high-affinity fractions by gradient salt elution of a heparin-Sepharose column . The distribution of the seven binding sites in the two fractions was determined in an immunoblotting assay using antibodies specific to each site, i.e . antibodies raised against synthetic peptide sequences found within each of the seven sites . The results of these two approaches demonstrate that site E and, to a somewhat lesser extent, site F bind to heparin with the highest affinity . Based on the analogy with apoE, in which the high-affinity heparin-binding site coincides with the domain of the protein that interacts with apoB,E (low density lipoprotein) receptors, the results of this study indicate that site E and site F, either singly or in combination, might constitute the receptor binding domain of apoB-100.

J Immunol, 1987 Aug 15, 139(4), 1135 - 41
B cell growth-promoting activity of recombinant human interleukin 4; Defrance T et al.; Human interleukin 4 (IL-4), also known as B cell stimulatory factor 1, is a T cell-derived glycoprotein consisting of 129 amino acids for which a cDNA has been recently isolated . IL-4 displays little or no B cell growth factor (BCGF) activity in the standard anti-IgM costimulatory assay using suboptimal concentrations of soluble anti-IgM antibody whereas the low m.w . BCGF is very active . When insolubilized anti-IgM was used as the costimulating agent, both IL-4 and the low m.w . BCGF were found to promote B cell proliferation . Human IL-4 is able to induce the proliferation of B lymphocytes preactivated for either 1 day with insolubilized anti-IgM antibody or for 3 days with Staphylococcus aureus strain Cowan I . However, IL-4 is poorly mitogenic for B cells preactivated for 1 day with the Staphylococcus strain whereas the low m.w . BCGF strongly enhances the proliferation of these B cells . These two findings demonstrate that the preactivation signal necessary to induce human B cells to proliferate in response to IL-4 is critical . The increased tritiated thymidine ({3H}dThd) uptake in preactivated B cell cultures with IL-4 reflects cel proliferation because cell cycle analysis demonstrates that IL-4 induces activated B cells to enter the S and G2/M phases of the cell cycle and the addition of IL-4 to preactivated B cell cultures permits the recovery of three- to fourfold more B cells after 4 days of culture . IL-4 and the low m.w . BCGF act in concert to induce the proliferation of anti-IgM-preactivated B cells as demonstrated by {3H}dThd uptake and cell cycle analysis . In striking contrast to the demonstrated antagonistic effect of interferon-gamma on the IL-4-induced expression of the low affinity receptor for IgE (Fc epsilon RL/CD23), on B cells, it was found that interferon-gamma enhanced the IL-4-induced proliferation of anti-IgM-preactivated B cells . Finally, it was found that IL-4 had to be present continuously during the culture period to exert an optimal growth-promoting effect on B cell blasts . As a conclusion, IL-4 is able to induce the proliferation of an appropriately activated subpopulation of human B cells.

J Immunol, 1987 Aug 15, 139(4), 1005 - 13
Comparative activation requirements of human peripheral blood, spleen, and lymph node B cells; Jelinek DF et al.; Human peripheral blood, spleen, and lymph node B cells were stimulated with Cowan I Staphylococcus aureus (SA) or F(ab')2 fragments of anti-mu antibody (anti-mu) and various lymphokines and were analyzed for proliferation and generation of Ig-secreting cells (ISC) . SA alone but not anti-mu stimulated minimal proliferation of each population . Recombinant IL 2 (r-IL 2) effectively promoted proliferation of SA-stimulated blood and spleen B cells, but supported less vigorous responses of lymph node B cells . By contrast, r-IL 2 enhanced DNA synthesis of all anti-mu-stimulated B cells early in culture, but did not promote sustained proliferation of anti-mu-stimulated lymph node B cells and only promoted ongoing DNA synthesis of some anti-mu-activated blood (eight out of 17) and spleen (five out of 14) B cell preparations . Recombinant interferon-gamma (r-IFN-gamma) and a commercial preparation of B cell growth factor (BCGF) also augmented DNA synthesis of all three B cell populations stimulated with SA or anti-mu early in culture, but neither alone was able to sustain maximal proliferation . Markedly enhanced sustained proliferation of all three anti-mu- and SA-stimulated B cell populations was noted when cultures were supported by the combination of r-IL 2 and BCGF, or to a lesser extent by r-IL 2 and r-IFN-gamma . The generation of ISC from SA-stimulated blood or spleen but not lymph node B cells was effectively supported by r-IL 2 alone . Differentiation of lymph node B cells required the combination of r-IL 2 and BCGF . These studies emphasize the importance of both the activation stimulus and the origin of the B cells in determining the lymphokine requirements of human B cell responsiveness.

J Immunol Methods, 1987 Aug 3, 101(2), 209 - 17
Selective removal of antigen-complexed IgG from cat plasma by adsorption onto a protein A-silica matrix; Snyder HW Jr et al.; The binding of normal cat IgG, heat-aggregated cat IgG and specific immune complexes (IC) containing cat IgG to a silica matrix containing covalently bound Staphylococcus aureus protein A was evaluated . The amounts of serum relative to protein A-silica, the flow rates and the perfusion times were representative of those existing when protein A-silica columns are used for therapeutic extracorporeal immunoadsorption of IgG and IC from humans and animals . When cat IgG was present in a large excess, approximately one molecule was bound to the matrix per molecule of solid-phase protein A with a KA of 1.5 X 10(6) 1/mol . Aggregated and immune complexed IgG bound to the matrix with relatively higher affinity . IC prepared in vitro between the purified envelope glycoprotein of the feline leukemia virus (FeLV gp70) and affinity-purified cat antibodies bound to the matrix even though normal IgG was present in greater than 10,000-fold excess . Once bound, IC were not eluted from columns upon further perfusion with normal serum . However, bound IgG was eluted from columns by further perfusion of normal serum or IC . IC were at least five-fold more efficient than normal IgG in exerting this effect . The results suggest that protein A-silica columns can be used for preferential removal of IC from plasma in a clinical or experimental setting.

J Antibiot (Tokyo), 1987 Aug, 40(8), 1184 - 92
HRE 664, a new parenteral penem . II . Evaluation of the pharmacokinetic behavior and the chemotherapeutic activity in animals; Klesel N et al.; The pharmacokinetic and chemotherapeutic properties of the new penem antibiotic HRE 664 (Fig . 1) were evaluated in experimental animals . High and sustained blood and serum levels were achieved following parenteral injection in mice, rats, dogs and monkeys . Half-lives ranged from 27 to 40 minutes in the various species tested . The antibiotic was well distributed in the rodents and penetrated well into tissues and body fluids . At 30 minutes after subcutaneous administration to mice (50 mg/kg), concentrations of between 12.4 and 35.9 micrograms/g were measured in the lungs, liver, heart and kidneys, that is 33 approximately 95% of the corresponding level in murine blood (37.7 micrograms/ml) . In experimentally induced infections in mice, HRE 664 displayed good chemotherapeutic activity particularly against septicemias caused by methicillin-sensitive and methicillin-resistant Staphylococcus aureus strains and on abscess formation induced by Bacteroides fragilis . Most of the cephalosporins and other beta-lactam antibiotics exhibited low efficacy against these strains of bacteria.

Acta Pathol Microbiol Immunol Scand {B}, 1987 Aug, 95(4), 213 - 7
Flucloxacillin in chronic leg ulcers . Penetration of flucloxacillin into chronic leg ulcer exudate and the effect on the bacteria; Sturup J et al.; The penetration of flucloxacillin into ulcer exudate was investigated in six patients with chronic leg ulcers . The flucloxacillin dosage used was 1 g orally three times daily for three days, and the serum and exudate concentrations were measured repeatedly during a 10 h-period following the first and the seventh dose . All the ulcers were contaminated with (S . aureus) Staphylococcus aureus either in pure culture (three ulcers) or in culture mixed with Gram-negative bacteria (three ulcers) . Bacterial counting in the ulcers was performed twice before and twice during the antibiotic treatment . The flucloxacillin concentrations measured in the ulcer exudate were found to be lower than the corresponding serum concentrations . However, the exudate concentrations were found to be above the minimum inhibitory concentration (MIC) for the contaminating S . aureus during an average of 7 h after each dose, and the number of S . aureus during the treatment period was reduced to less than 0.01% of the initial number . The Gram-negative bacteria were not susceptible to flucloxacillin . The number of these bacteria decreased before flucloxacillin treatment but increased again during treatment, probably owing to the changed conditions in the ulcers following the marked decrease in the number of S . aureus.

Acta Orthop Scand, 1987 Aug, 58(4), 361 - 4
Cefuroxime prophylaxis in trochanteric hip fracture operations; Hedstrom SA et al.; In a double-blind randomized study of antibiotic prophylaxis in trochanteric fractures operated on with a nail and plate, a 24-hour intravenous administration of cefuroxime 0.75 grams thrice daily (Group B) was compared with the previous regimen of cefuroxime for 24 hours plus 6 days of oral cephalexin (Group A) . In each group, 56 (Group A) and 65 (Group B) patients could be evaluated . One deep infection occurred in Group B with growth of Staphylococcus aureus, and another 4 patients had discharge and cultures of which 2 showed S . aureus . In Group A, 6 patients had signs of an infection, and in 3 patients cultures were taken but were negative . There were no differences between the groups . We concluded that the prophylaxis time need not be longer than 3 days.

Acta Orthop Scand, 1987 Aug, 58(4), 354 - 60
Tissue vitality in septic gonitis . 99mTc-DPD scintimetry in puppies; Knudsen VE et al.; After a single intraarticular injection of 10(9) Staphylococcus aureus in 12 puppies, septic arthritis developed in all the experimental knees after 48 hours . A considerable variability in scintigraphic appearance was observed . The juxtaarticular growth plates showed either unchanged or slightly decreased uptake except in 1 dog exhibiting a definite increase in tracer uptake . The epiphyseal uptake showed no consistent pattern . The intraarticular pressure of the arthritic joints increased significantly, but was not related to the tracer uptake pattern . We conclude that delayed joint scintigraphy as a single investigation in early septic arthritis does not provide diagnostic information and may be misleading.

Acta Orthop Scand, 1987 Aug, 58(4), 351 - 3
Staphylococcal adherence to chicken cartilage; Nade S et al.; Immature chicken cartilage was incubated in a Staphylococcus aureus suspension and then washed . Scanning electron microscopy and radiolabel measurements showed increased adherence to cartilage with increasing bacterial concentration . Preheating of the bacteria did not reduce the adherence property, but trypsin treatment did.

J Clin Pathol, 1987 Aug, 40(8), 837 - 40
Protein A and coagulase expression in epidemic and non-epidemic Staphylococcus aureus; Roberts JI et al.; Strains of Staphylococcus aureus were divided into groups on the basis of antimicrobial sensitivity and epidemiology and tested for protein A expression in a simple microtitre test, which detected the non-immunological binding of immunoglobulin to protein A on whole cells of S aureus . Isolates of the methicillin resistant strain prevalent in south east England (EMRSA) showed a low expression of protein A compared with the other strains of methicillin resistant S aureus (MRSA), other multiple resistant strains, and sensitive strains . Protein A and coagulase expression in 27 strains of MRSA from 15 countries associated with hospital outbreaks were compared with 27 strains of MRSA from 11 countries reported to be sporadic isolates . Twenty four of the 27 outbreak associated MRSA showed low expression of protein A and high expression of coagulase . Conversely, sporadic strains generally gave higher levels of protein A and a wide variety of coagulase reactions . The results suggest that many epidemic strains of MRSA may have phenotypic characteristics that distinguish them from sporadic strains.

Infect Control, 1987 Aug, 8(8), 325 - 8
Staphylococcus aureus mediastinitis: prognostic usefulness of an early medicosurgical therapy; Voiriot P et al.; Six cases of acute Staphylococcus aureus mediastinitis after median sternotomy were reported . Five resulted from an asymptomatic disseminator of S aureus present in the operating room . Each case was characterized by an acute bacteremic phase, occurring after a mean interval of 8.2 +/- 1.7 days after the surgical procedure; within 24 to 36 hours all patients had a temperature above 39 degrees C, toxic appearance, and marked leukocytosis . Pericicatricial inflammation was moderate, instability of the sternum was present in only two patients, and chest roentgenogram was not helpful in making an early diagnosis . No risk factor for mediasinitis in connection with the perioperative or postoperative periods was noted in cases compared with a control group of 103 patients . All strains of S aureus were susceptible in vitro to the antibiotic regimen used in prophylaxis . All patients underwent early surgical reopening of the mediastinum within 47 +/- 15 hours after the first sign of acute mediastinitis . Mediastinal debridement and continuous irrigation-suction with dilute povidone-iodine solution were associated with intravenous antistaphylococcal therapy for a period of four to six weeks . All patients survived and no recurrence was observed, a finding we think due to early diagnosis and aggressive medicosurgical therapy.

J Clin Microbiol, 1987 Aug, 25(8), 1446 - 9
Production of toxic shock syndrome toxin 1 by Staphylococcus aureus as determined by tampon disk-membrane-agar method; Robbins RN et al.; The influence of 17 commercially available tampons on production of toxic shock syndrome toxin 1 (TSST-1) by Staphylococcus aureus was investigated by using a tampon disk method . Filter membranes overlaying agar medium (with or without blood) in small petri dishes were spread inoculated with a TSST-1-producing strain of S . aureus . Disks cut from unrolled tampons were pressed and laid on the inoculated membranes; incubation was for 19 h at 37 degrees C with 5% CO2 in air . CFU on the membranes and in the disks were enumerated, and the presence of TSST-1 in the disks and in the agar layers was determined . Tampons made of different materials supported characteristic levels of cell growth and toxin production in the tampon . Colonization of the interface surface of the tampon disks was heavy . The number of CFU extracted from the tampon disks ranged from 5 X 10(10) to 82 X 10(10) . There was little variation in the CFU recovered from the membranes ({1.9 +/- 0.4} X 10(11)) . Sixty to 170 micrograms of TSST-1 was recoverable from the agar, with an additional 10 to 90 micrograms recoverable from tampon disks, depending on the type of tampon disk . The amount of toxin in the agar layer from the various tampon disks was relatively constant and indicated an important contribution of toxin from vaginal S . aureus cells not growing in the tampon . The main role of tampons in toxic shock syndrome may be that of providing a fibrous surface for heavy colonization and sufficient air for TSST-1 production.

Cell Struct Funct, 1987 Aug, 12(4), 395 - 9
Extracellular products of Staphylococcus aureus reversibly inhibit the terminal differentiation of cultured mouse epidermal cells; Sugai M et al.; The effect of extracellular products from Staphylococcus aureus on the differentiation of mouse epidermal cells was studied using an in vitro cell culture system . The extracellular products from a clinical strain of S . aureus isolated from human skin lesions reversibly inhibited the Ca++-induced terminal differentiation of epidermal cells, as determined by their morphology and the extent of cornified envelope formation . This suggests that a similar modification of cell differentiation is involved in the pathogenesis of S . aureus-induced skin disease.

Am Rev Respir Dis, 1987 Aug, 136(2), 293 - 7
Alveolar macrophage dysfunction in systemic lupus erythematosus; Wallaert B et al.; A high frequency of pulmonary infections has been a well-described feature of systemic lupus erythematosus (SLE) . Alveolar macrophages (AM) play a crucial role in pulmonary bacterial defense . We therefore examined the antibacterial activity of AM and generation of superoxide anion in 17 patients with SLE without clinical or radiologic pulmonary changes and in 8 control subjects . Total cell count and cellular viability of AM (trypan blue exclusion) did not differ significantly between patients and control subjects . Antibacterial activity v/s Staphylococcus aureus was significantly decreased in both untreated and corticosteroid-treated patients (respectively, -16.2 +/- 7.4 and -42 +/- 12% compared with the normal value of 51 +/- 12%, p less than 0.001) . The defect of antibacterial activity was observed as well v/s S . aureus as v/s Escherichia coli . In contrast, chemiluminescence response of AM before and after stimulation by either phorbol myristate acetate or opsonized zymosan did not differ among control subjects and treated and untreated patients with SLE . We did not find any correlation between disease activity and AM function . Antibacterial activity of normal AM was shown to be significantly reduced by previous incubation with SLE serum compared with normal human serum . Thus, our findings suggest that alteration of antibacterial activity of AM may contribute to the increased susceptibility to lung infections observed in SLE.

Am J Clin Pathol, 1987 Aug, 88(2), 231 - 5
Variation from standards in Staphylococcus aureus susceptibility testing; Pfaller MA et al.; In an effort to assess the degree of methodologic variation and adherence to current guidelines for detection of methicillin-resistant Staphylococcus aureus (MRSA), the authors surveyed the susceptibility testing practices of all 162 microbiology laboratories in the Veterans Administration (VA) system . Completed questionnaires were returned by 136 (84%) of the laboratories . Overall, 96 (71%) laboratories used disk diffusion testing, 54 (40%) used manual broth dilution, and 36 (26%) used an automated method . The percentage of MRSA detected ranged from 0 to 52%, with a mean of 10% . In general, fewer than 60% of laboratories followed the current susceptibility testing guidelines for key methodologic variables such as inoculum preparation, duration of incubation, and medium supplementation . Failure to adhere to these guidelines may result in suboptimal detection of MRSA.

J Med Microbiol, 1987 Aug, 24(1), 75 - 81
Environmental factors affecting toxic shock syndrome toxin-1 (TSST-1) synthesis; Sarafian SK et al.; The production of toxic shock syndrome toxin-1 (TSST-1) was studied in batch and continuous culture of Staphylococcus aureus strain 1169 in a carbohydrate-free chemically defined medium (CDM) . In continuous culture oxygen- and arginine-limitation were required for steady-state TSST-1 synthesis . Aeration suppressed toxin synthesis . The amount of TSST-1 per mg dry weight (specific toxin) at dilution rates from 0.05 to 0.15 h-1 was inversely proportional to the dilution rate . Protease activity increased with increasing dilution rates . In batch culture, TSST-1 began to accumulate in the medium towards the end of the exponential phase of growth, after a critical cell mass was attained . Maximum specific toxin production was observed in medium with an initial pH between 6.5 and 7.0 . Growth and toxin synthesis took place in anaerobic conditions when CDM was supplemented with pyruvate and uracil . The Mg++ concentration had no effect on the specific toxin in anaerobic conditions . In aerobic conditions, specific toxin increased c . 23-fold as the Mg++ concentrations increased to 0.4 mM . Further increases in the Mg++ concentration resulted in a reduction in specific toxin.

Epidemiol Infect, 1987 Aug, 99(1), 191 - 200
Characterization of non-typable strains of Staphylococcus aureus from cases of hospital infection; Vindel A et al.; A high percentage of non-typable (NT) Staphylococcus aureus strains was isolated in Spanish hospitals during 1984 and 1985 . Several alternative methods of typing were employed to study these isolates . These were: phage-typing at 1000 X RTD, phage-typing after heat-treatment (48 degrees C), thermal shock (56 degrees C), reverse-typing and induction of additional phages . Using these methods the number of NT isolates was reduced by 60% . Best results were obtained with heat-treatment . Additional phages and reverse-typing were also useful . A scheme for the study of outbreaks and sporadic cases caused by NT strains is proposed using the methods described.

Immunology, 1987 Aug, 61(4), 497 - 502
C-kinase activity in normal B cells treated with Staphylococcus aureus, Cowan strain I, and phorbol ester: response differences in a patient with prolymphocytic leukaemia; Cooper R et al.; Ca+2/phospholipid-dependent kinase (C-kinase) activity is intimately involved with the B-cell response after ligation of its mIg receptor . Here, we extend previous findings with anti-mIg by showing that polyclonal stimulation with Staphylococcus aureus, Cowan strain I (SAC), could also translocate C-kinase from the cytosol of intact, normal human B lymphocytes . This stimulus could not, however, induce redistribution of enzyme in a prolymphocytic leukaemia (PLL) B-cell population . Despite a normal density of mIg receptors on the latter cell type, PPL cells could not be stimulated to proliferate, express interleukin-2 (IL-2) receptors, and differentiate under the influence of SAC, even in the presence of exogenous IL-2 . Normal B cells could respond appropriately to the same stimuli . PLL cells had a normal content of C-kinase activity and the enzyme could be translocated under the influence of the specific agonist, 12-0-tetradecanoyl phorbol-13-acetate (TPA) . The same agent could induce cellular proliferation and differentiation in PLL cells, showing that its terminal C-kinase dependent pathways were intact . We conclude that the mIg receptor mechanism of this PLL population could not activate C-kinase appropriately.

Acta Pathol Microbiol Immunol Scand {C}, 1987 Aug, 95(4), 167 - 72
Recombinant interleukin 2 induces proliferation and differentiation of human B lymphocytes; Punnonen J et al.; Highly purified normal human tonsillar B lymphocytes were cultured with recombinant IL 2 (rIL 2) . The responses were minimal when the cells were cultured with rIL 2 alone . Dose-dependent proliferation and differentiation into immunoglobulin-secreting cells were found when the cells were simultaneously activated with Staphylococcus aureus Cowan I (SAC) . T cells were not found to proliferate in these cultures . B-cell responses were not augmented when 16% T cells were added to the B-cell population . In contrast, B-cell proliferation and Ig secretion were significantly enhanced by adding 0.5% T cells when cultured in the presence of pokeweed mitogen (PWM) . Moreover, B-cell stimulation by rIL 2 was detected at very low cell densities . It is concluded that IL 2 directly affects the function of activated B cells.

Chemioterapia, 1987 Aug, 6(4), 261 - 3
In vitro susceptibility of methicillin-resistant Staphylococcus aureus to imipenem and other antimicrobial agents; Ishag AJ et al.; Minimum inhibitory (MIC) and minimum bactericidal (MBC) concentrations of imipenem, pefloxacin, BMY 28142, were compared with those of vancomycin and other established antistaphylococcal antimicrobial agents, rifampicin, fusidic acid, clindamycin, tobramycin and amikacin against 50 clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA) . Imipenem was more active than the other tested antimicrobial agents except for vancomycin which was the most active . The MIC90 of imipenem was 2 micrograms/ml, while the MBC90 was 4.2 micrograms/ml and in all cases the MBCs were only 2-4 times higher than the MICs . Pefloxacin had significant bacteriostatic and bactericidal effect against the majority of the isolates, with an MIC90 of 12.5 micrograms/ml (range 0.4-25 micrograms/ml) and in most cases the bactericidal concentrations were 2-4 times the MIC, with a few isolates having MBCs 8 or more times the MICs . The results obtained confirm that imipenem has excellent in vitro activity against locally isolated MRSA.

Jpn J Antibiot, 1987 Aug, 40(8), 1364 - 76
{Bacteriological and clinical studies on flomoxef in the pediatric field}; Sunakawa K et al.; Bacteriological and clinical studies on flomoxef (FMOX, 6315-S) were performed and the results obtained are summarized below . 1 . The MIC values of FMOX against 307 clinically isolated strains of Staphylococcus aureus were 0.024 to 100 micrograms/ml with a peak MIC of 0.39 microgram/ml, and the MIC90 value was 1.56 micrograms/ml . The MIC90 against methicillin resistant S . aureus (MRSA) was 25 micrograms/ml . 2 . FMOX was administered to 15 children with pediatric bacterial infections, and the effectiveness was rated excellent or good in all cases . 3 . In bacteriological evaluation, 7 of 11 strains identified prior to the treatment were eliminated (63.6%) . 4 . As side effects, diarrhea or soft stool was found in 3 cases and eruption in 1 case . As abnormal laboratory values, eosinophilia and thrombocytosis were found in 1 case each . 5 . On the intestinal bacterial flora, FMOX had a marked influence just as did other Group 4 and 5 cephems antibiotics . 6 . FMOX interfered little with the coagulation system or platelet aggregation.

Zh Mikrobiol Epidemiol Immunobiol, 1987 Aug, (8), 85 - 8
{Detection of antibodies to preparations of Staphylococcus aureus peptidoglycan, teichoic acids and polysaccharide in patients' sera}; Vaneeva NP et al.; Enzyme immunoassay systems on the basis of S . aureus teichoic acids, peptidoglycan and polysaccharide-containing preparation are incapable of reliably diagnosing the staphylococcal nature of postoperative complications in oncological patients.

Surgery, 1987 Aug, 102(2), 402 - 8
Does bacteremia pose a direct threat to synthetic vascular grafts?
White JV, Freda J, Kozar R, Serfass D, Cundy K, Comerota AJ, Ritchie WP.
This study was undertaken to determine the significance of graft lumen exposure to blood-borne organisms in the development of graft infection . Three groups of dogs were studied . In group I (n = 20), the infrarenal aorta was dissected from surrounding tissue, divided, and reconstructed with a Dacron tube interposition graft . In group II (n = 9) the aorta was similarly isolated, but Dacron graft material was wrapped around the intact aorta . In group III (n = 13) the infrarenal aorta was isolated, but no graft material was placed . All dogs were given intravenous 1 X 10(7) Staphylococcus aureus at the completion of surgery . Group I grafts were harvested 8 hours, 1 day, or 21 days after bacterial challenge . Group II and III grafts were harvested 1 day or 21 days after infusion . At the time of harvest, selective cultures of the periaortic tissue (PAT), periaortic graft (PAG), and interposition graft lumen (GL) were taken . The overall infection rates were similar, with 17 of 20 (85%) dogs in group I, 6 of 9 (67%) in group II, and 11 of 13 (85%) in group III found to be culture positive . In group I, 3 of 3 dogs at 8 hours, 2 of 2 on day 1, and 12 of 15 on day 21 had positive PAT cultures . Only 4 of 15 on day 21 had positive GL cultures . In group II, 4 of 5 dogs on day 1 and 2 of 4 on day 21 had positive PAT and PAG cultures . In group III, 9 of 9 animals on day 1 and 2 of 4 on day 21 had positive PAT cultures . All aortic lumen cultures were negative in groups II and III . The difference between GL and PAT cultures was statistically significant in all groups (I, p = 0.01; II, p = 0.05; III, p = 0.01) . Serial quantitative blood cultures revealed a mean bacterial load of 10.5 +/- 4.5 CFU/ml at 15 minutes postinfusion, which fell steadily until no bacteria were detected at 3.5 hours . Lymphangiography demonstrated periaortic pooling of lymph in the immediate postoperative period . These data suggest that the bacteremia in this model is transient and rapidly clears . Periaortic tissues quickly sequester bacteria, possibly because of lymphatic leakage . The GL appears to be secondarily infected.

Proc Natl Acad Sci U S A, 1987 Aug, 84(16), 5615 - 9
Cloning of the alpha chain of human platelet glycoprotein Ib: a transmembrane protein with homology to leucine-rich alpha 2-glycoprotein; Lopez JA et al.; Glycoprotein Ib is a surface membrane glycoprotein of platelets that functions as a receptor for von Willebrand factor . It is a heterodimer composed of an alpha and a beta chain linked by a disulfide bond(s) . A phage lambda gt11 cDNA expression library prepared from mRNA from a human erythroleukemia cell line, HEL, was screened using an affinity-purified antibody to the glycocalicin portion of the alpha chain of glycoprotein Ib . Eleven positive clones were isolated and plaque-purified . The largest cDNA insert was 2420 nucleotides in length and coded for a leader sequence of 16 amino acids, a mature protein of 610 amino acids, and a stop codon . It also contained 42 nucleotides of 5' noncoding sequence and 497 nucleotides of 3' noncoding sequence, including a poly(A) tail . The amino acid sequence of the alpha chain of GPIb predicted from the cDNA agreed completely with the sequence of 156 amino acids that was determined by Edman degradation of peptides isolated from human platelet glycocalicin after digestion with trypsin or Staphylococcus aureus V8 protease . The extracytoplasmic domain of the alpha subunit of GPIb contains several noteworthy structural features, including a region of seven tandem repeats of 24 amino acids that are homologous with those present in leucine-rich alpha 2-glycoprotein . The extracytoplasmic domain also contains two hydrophilic regions, one rich in charged amino acids and a second rich in serine and threonine residues . The region rich in serine and threonine includes five repeats of nine amino acids as well as the majority of the O-linked carbohydrate sites present in the molecule . The extracytoplasmic domain is followed by a potential transmembrane segment of approximately 29 amino acids and a potential intracellular domain of approximately 100 amino acids located at the carboxyl end of the molecule.

Acta Pathol Microbiol Immunol Scand {B}, 1987 Aug, 95(4), 253 - 5
Influence of carbon-dioxide tension and medium buffer concentration on medium pH and MIC values of erythromycin for Escherichia coli ATCC 25922 and Staphylococcus aureus ATCC 25923 in a micro-aerobic atmosphere; Andreasen JJ et al.; The effect of an elevated carbon-dioxide tension on medium pH and MIC determination with erythromycin for Staphylococcus aureus (ATCC 25923) and Escherichia coli (ATCC 25922) was investigated in a micro-aerobic atmosphere, using an agar dilution method . During 30 h of incubation in an atmosphere containing approximately 6% CO2, 5% O2 and 89% N2, pH of uninoculated, as well as of seeded, blood agar plates decreased from 7.4 to 6.8 . MIC values of erythromycin were two dilution steps higher after micro-aerobic incubation than after aerobic incubation . A decrease in agar pH during micro-aerobic incubation was eliminated by increasing the phosphate buffer concentration in the medium . However, MIC values of erythromycin remained high.

J Am Acad Dermatol, 1987 Aug, 17(2 Pt 1), 246 - 50
Treatment of junctional epidermolysis bullosa with epidermal autografts; Carter DM et al.; We have successfully treated chronic facial erosions in three boys with junctional epidermolysis bullosa . In each patient, keratinocytes were harvested from the roof of suction blisters created on clinically uninvolved skin . They were grown in tissue culture on collagen sponges and grafted onto facial erosions that were previously treated with 2% mupirocin ointment . This experimental antibiotic ointment has proved efficacy in eradicating cutaneous pathogens such as Staphylococcus aureus from chronic wounds . In two patients, complete reepithelialization was achieved over 7 and 10 months, respectively, and partial reepithelialization occurred in another patient in whom treatment is ongoing . Epidermal autografts are a promising means for improving function and appearance in eroded skin caused by junctional epidermolysis bullosa.

Cell Immunol, 1987 Aug, 108(1), 97 - 108
Role of HLA class I and class II antigens in activation and differentiation of B cells; Giudizi MG et al.; The monoclonal antibodies (MoAb) CR10-214, CR11-115, and Q1/28 to distinct monomorphic determinants of HLA class I antigens, the MoAb CL413 and PTF29.12 recognizing monomorphic determinants of HLA-DR antigens, the anti-HLA-DQw1 MoAb KS11, the anti-HLA-DPw1 MoAb B7/21, and the anti-HLA-DR,DP MoAb CR11-462 were tested for their ability to modulate human B-lymphocyte proliferation and maturation to IgM-forming cells . Purified tonsillar B cells were stimulated with Staphylococcus aureus bacteria of the Cowan first strain (SAC) or anti-human mu-chain xenoantibodies, as well as in growth factor- or T-cell-dependent activation cultures . The B-cell proliferative responses induced by SAC or by mitogenic concentrations of anti-mu-chain xenoantibodies were inhibited by some of the anti-HLA class I and anti-HLA class II monoclonal antibodies tested . The same antibodies were effective inhibitors of the proliferation of B cells stimulated with interferon-gamma (IFN-gamma) or interleukin-2 (IL-2) and with submitogenic concentrations of anti-mu-chain xenoantibodies . The proliferation induced by IL-2 of SAC-preactivated B cells was inhibited by some of the anti-HLA class II monoclonal antibodies, but not by the anti-HLA class I monoclonal antibodies tested . This inhibition appeared to reflect at least in part a direct effect on later events of the B-cell activation cascade, since some anti-HLA class II monoclonal antibodies still exerted considerable inhibitory activity when added together with IL-2 to SAC-preactivated B cells after the third day of culture . Anti HLA-DR, DQ, and DP monoclonal antibodies consistently inhibited the IgM production induced in B cells by T cells alone, T cells plus pokeweed mitogen (PWM), SAC plus IL-2, or IL-2 alone . In contrast, two of the three anti-HLA class I monoclonal antibodies tested inhibited the IgM production in cultures stimulated with SAC plus IL-2 and one the IgM production induced by IL-2 alone, but none of them had inhibitory effects on T-cell dependent IgM production . The results reported herein indicate that HLA class II molecules directly participate in different phases of the B-cell activation cascade . In addition, our data also suggest that HLA class I molecules can be involved in the events leading to B-cell proliferation and differentiation into immunoglobulin-secreting cells.

J Gen Virol, 1987 Aug, 68 ( Pt 8), 2079 - 92
Herpes simplex virus replication and protein synthesis in a human blood-derived cell line; Soslau G et al.; Herpes simplex virus (HSV) types 1 and 2 were shown to replicate in a newly described human cell line (Meg) derived from the peripheral blood of a healthy volunteer . The cell line has both megakaryocyte-like and B cell-like properties . Upon infection with HSV-1 or -2, at a m.o.i . between 0.5 and 5, unlike B and T cells, the Meg cells were growth-arrested and this was accompanied by cytopathic effects and virus replication . The HSV proteins and glycoproteins B and D (gB and gD) made in the blood-derived Meg cells were compared to the corresponding proteins made in the non-blood-derived cell lines, Vero (African green monkey kidney cell) and HEp-2 (human epidermoid carcinoma cell) . The maximum level of HSV protein synthesis occurred earlier in the Meg cells than in the Vero and HEp-2 cells . The electrophoretic pattern of HSV-1 and -2 proteins made in the Meg cell line was similar to the corresponding proteins made in the Vero and HEp-2 cell lines; however, some qualitative and quantitative differences were evident . There were no apparent differences detected in the migration pattern of gB made in all three cell lines while significant differences were observed with the gD species . However, upon hydrolysis with Staphylococcus aureus V-8 protease of the monoclonal antibody-purified gB and gD, distinct differences were observed in the electrophoretic pattern of the generated peptide fragments of both gB and gD made in the three cell lines . The results demonstrate that a human blood cell can support HSV replication and that species-specific post-translational modification of gB and gD occurs in HSV-infected Vero cells as compared to HSV-infected human cells.

Zentralbl Bakteriol Mikrobiol Hyg {A}, 1987 Aug, 266(1-2), 127 - 36
Correlation between enterotoxigenicity, tested by different ELISA-techniques, antibiotic resistance patterns and phage groups of Staphylococcus aureus strains; Pickenhahn P et al.; A group of 596 Staphylococcus aureus strains isolated from various clinical sources or implicated in food poisoning was investigated for enterotoxins A and B (SEA and SEB) production . The conventional ELISA techniques (competitive and sandwich ELISA) were compared with a newly developed avidin-biotin ELISA in their ability to detect the enterotoxins . The avidin-biotin system was not remarkably influenced by SPA up to 10 micrograms/ml . A semi-quantitative competitive ELISA for the detection of staphylococcal protein A (SPA) in culture supernatants was carried out in parallel . The strains isolated in cases of food poisoning showed different antibiotic resistance patterns, whereas the strains from clinical sources were selected for either methicillin or penicillin resistance only . The strains isolated in food poisoning outbreaks (FP strains) were enterotoxin A positive in 22%, enterotoxin B positive in 11%, and SEA + SEB positive in 9% of cases . The strains with resistance to penicillin only (PER strains) produced SEB in 26%, SEA in 14%, and both toxins in 7% of the cases . The methicillin-resistant strains (MCR strains) produced SEA in 59% of cases, whereas SEB was produced in 6% only (SEA + SEB: 20%) . 37% of the SEA producers belonged to phage group III (SEB: 30%; SEA + SEB: 25%) and 12% (SEB: 11%; SEA + SEB: 9%) to phage group I . 26% of the SEA-producing and 37% of the SEB-producing strains (SEA + SEB: 23%) were non-typable.

Clin Exp Immunol, 1987 Aug, 69(2), 358 - 67
In vitro IgM and IgM rheumatoid factor production in response to Staphylococcus aureus Cowan I: evidence for the role of Leu-2-positive T suppressor cells and radiosensitive T helper cells; Goldstein R et al.; Staphylococcus aureus Cowan I (SAC) is a potent stimulus of B cell proliferation and differentiation, the latter being T cell-dependent . It has been suggested that immunoglobulin and IgM rheumatoid factor (RF) production in response to SAC involves radiosensitive T helper cells . We studied normal peripheral blood mononuclear cell (PBMC) cultures to assess the roles of radiosensitive T cells and Leu-2 positive suppressor cells in the cellular control of SAC-stimulated IgM and IgM RF responses . Depletion of Leu-2 positive T cells from reconstituted autologous PBMC cultures resulted in an increase in SAC-stimulated IgM production in the majority of individuals, implying the involvement of Leu-2 positive suppressor T cells in this system . Suppression by Leu-2 positive cells is less evident in the SAC-induced IgM RF response, suggesting qualitative differences between IgM and IgM RF SAC-stimulated responses in PBMC cultures from the same normal individuals . Irradiation (1000 rads) of the T cell-enriched subpopulation, either with or without Leu-2 positive cell depletion, resulted in statistically significant decreases in IgM and IgM RF production in response to SAC in reconstituted autologous cultures, providing further evidence of a Leu-2 negative radiosensitive sheep-cell rosetting cell active in in vitro SAC responses . In contrast, PWM-stimulated PBMC cultures showed almost exclusively increases in total IgM and IgM RF production when T cells were irradiated (1000 rads) before culture, consistent with the radiosensitive T suppressor cell involved in the in vitro immunoglobulin responses to PWM . The same five out of nine individuals produced IgM RF, under different culture conditions, in response to PWM and SAC, suggesting that the ability of an individual to produce IgM RF lies intrinsically within the B cell.

J Virol Methods, 1987 Aug, 17(1-2), 159 - 74
A sensitive enzyme-linked immunosorbent assay for the detection of herpes simplex virus antigens; Middeldorp JM et al.; A sandwich ELISA for the detection of herpes simplex virus (HSV) antigens was developed using sheep anti-HSV F(ab')2 fragments for capture and an indirect antibody system for detection . Current detection limits are 0.5 ng protein for HSV1 and 1.5 ng protein for HSV2 . This compares to a single HSV1-infected Vero-cell in a background of 10(6) non-infected cells or 10 plaque forming units (PFU) of HSV1 in culture supernatants as determined in separate experiments . Limiting dilution experiments show that one PFU of HSV1 can be detected after overnight culture in both supernatant and cell extracts . The use of F(ab')2 for capture completely eliminated binding of Staphylococcus aureus . No cross-reactivity was observed with other human herpes viruses . When evaluated with 245 random 'left-overs' of genital swab specimens in transport medium the test showed a sensitivity and specificity of 77.2 and 97.8%, respectively, with respect to virus isolation in culture . In a preliminary study on 16 direct ELISA swab-specimens extracted in 0.5 ml ELISA sample buffer both sensitivity and specificity were 100% with respect to culture . In both clinical series there was a proportional relationship between the ELISA value and the estimated amount of infectious virus in the specimen.

EMBO J, 1987 Aug, 6(8), 2351 - 7
Cloning and expression of the gene for a fibronectin-binding protein from Staphylococcus aureus; Flock JI et al.; The gene encoding the fibronectin-binding protein (FNBP) from Staphylococcus aureus strain 8325-4 was isolated from a gene bank in pBR322 . The original clone, containing a 6.5-kb insert, gave a functional product present in the periplasm of Escherichia coli . Analysis of polypeptides isolated after affinity chromatography on fibronectin-Sepharose followed by ion-exchange chromatography revealed two gene products, 87 and 165 kd in mol . wt . The amino acid compositions of these two polypeptides and a native FNBP from S . aureus strain Newman were very similar . Antibodies raised against the native FNBP from strain Newman precipitated the 125I-labelled 165-kd polypeptide, and unlabeled 165- and 87-kd polypeptides as well as native FNBP inhibited the immunoprecipitation reactions . The region of the fnbp-gene encoding the fibronectin-binding activity has been identified and subcloned in an expression vector based on the staphylococcal protein A gene . The resulting product in E . coli is an extracellular fusion protein consisting of two IgG-binding domains of protein A followed by a fibronectin-binding region . The fusion protein binds to fibronectin and completely inhibits the binding of fibronectin to intact cells of S . aureus.

J Biol Chem, 1987 Jul 25, 262(21), 10384 - 91
Structure of the lipid moiety of the Leishmania donovani lipophosphoglycan; Orlandi PA Jr et al.; The lipid moiety of the lipophosphoglycan of Leishmania donovani had been isolated and characterized as a novel lyso-alkylphosphatidylinositol . Treatment of lipophosphoglycan with either 10% NH4OH or a phosphatidylinositol-specific phospholipase C from Staphylococcus aureus liberated a monoalkylglycerol substituent . Structural characterization of the monoalkylglycerol by gas-liquid chromatography-mass spectrometry indicated the presence of two saturated, unbranched hydrocarbons: a C24 alkyl chain comprising 78% of the lipid with the remaining 22% as a C26 alkyl chain . Periodate sensitivity demonstrated that the alkyl side chain is linked to the C-1 position of the glycerol backbone . Treatment of lipophosphoglycan with nitrous acid released 1-O-alkylglycerophosphorylinositol due to an unacetylated glucosamine residue linked to the inositol of the lyso-alkylphosphatidylinositol . Quantitative analysis of the organic solvent-soluble product of nitrous acid deamination of lipophosphoglycan confirmed the expected ratio of inositol:phosphate:1-O-alkylglycerol as 1:1:1 . These results suggest that L . donovani anchors its lipophosphoglycan with a unique lipid component.

J Biol Chem, 1987 Jul 25, 262(21), 10164 - 70
The isolation and characterization from rabbit reticulocytes of two forms of eukaryotic initiation factor 2 having different beta-polypeptides; Dholakia JN et al.; We have isolated from the high salt wash of rabbit reticulocyte ribosomes two forms of the polypeptide chain initiation factor 2 (eIF-2) which differ with respect to their beta-subunit, GDP content, and sensitivity to Mg2+ in ternary (eIF-2 X GTP X Met-tRNAf) and binary (eIF-2 X GDP) complex formation . The form of eIF-2 eluting first from a cation exchange (Mono S, Pharmacia) column has a beta-subunit of lower molecular weight (eIF-2(beta L} and a more acidic pI value than the form eluting at a higher salt concentration (eIF-2(beta H} . These two forms of eIF-2 beta-polypeptides are also detected in reticulocyte lysates when the proteins are resolved by two-dimensional isoelectric focusing-dodecyl sulfate polyacrylamide gel electrophoresis followed by immunoblotting . The peptide mapping of the isolated beta-subunits after limited proteolysis by papain, pancreatic protease, alpha-chymotrypsin, or Staphylococcus aureus V8 protease further demonstrates that the two forms of beta-subunits are not the product of a non-specific proteolytic action that occurred during the purification procedure, but rather reflects the existence in vivo of both forms of eIF-2 . The GDP content of eIF-2(beta L) and eIF-2(beta H) is approximately 0.85 and 0.22 mol of GDP/mol of eIF-2, respectively . The KD for GDP of eIF-2(beta L) was lower (2.2 X 10(-9) M) than that of eIF-2(beta H) (6.0 X 10(-8) M) . In the presence of 1 mM Mg2+, the activities of eIF-2(beta L) and eIF-2(beta H) in forming a binary and a ternary complex are inhibited 90 and 25%, respectively . The extent of Mg2+ inhibition and its reversal by the guanine nucleotide exchange factor is directly proportional to the amount of GDP bound to eIF-2 . No inhibition by Mg2+ is observed when eIF-2-bound GDP is removed by alkaline phosphatase . In the presence of the guanine nucleotide exchange factor, both forms of eIF-2 are equally active in ternary complex formation, and the complex formed is quantitatively transferred to 40 S ribosomal subunits.

J Immunol Methods, 1987 Jul 16, 101(1), 29 - 36
Distinct culture requirements for activation and proliferation of human peripheral blood B lymphocytes by Staphylococcus aureus Cowan strain and growth factors; Choi YS et al.; The concentration of fetal calf serum (FCS) and the culture period were the crucial culture conditions in measuring B cell growth factor (BCGF) and IL-2 activity in vitro . Higher concentrations of FCS (10-15%) significantly inhibited BCGF activity; whereas, lower concentrations of FCS (less than 2.5%) were not sufficient for the response to IL-2 . Kinetic experiments showed that the culture period for BCGF should be shorter than that for IL-2, while BCGF in combination with IL-2 induced a synergistic proliferation of B cells in a longer culture period . Adding BCGF after 48 h of SAC stimulation reduced the reaction . Hence, the conventional method using preactivated B cells does not measure BCGF but mostly IL-2 . Furthermore, minute amounts of BCGF activity can be more sensitively determined by co-culturing with fixed amounts of IL-2.

Tijdschr Diergeneeskd, 1987 Jul 15, 112(14), 844 - 6
{Staphylococcal enterotoxemia following consumption of a pork fricandeau}; Landheer JE et al.; A case of food poisoning in a 45-year-old man, caused by enterotoxin C produced by Staphylococcus aureus following the consumption of porc-fricandeau is reported . The findings are discussed on the basis of data from the literature on abscesses in slaughtered animals.

Eur J Biochem, 1987 Jul 15, 166(2), 357 - 63
Structure and regulation of eukaryotic initiation factor eIF-2 . Sequence of the site in the alpha subunit phosphorylated by the haem-controlled repressor and by the double-stranded RNA-activated inhibitor; Colthurst DR et al.; Eukaryotic protein synthesis initiation factor 2 (eIF-2) can be phosphorylated on its alpha subunit by two well-characterised protein kinases, termed the haem-controlled repressor (HCR) and the double-stranded RNA-activated inhibitor (dsI) . Phosphorylation of eIF-2 by these kinases is thought to be important in the regulation of peptide-chain initiation . We report the location of the serine residue in the alpha subunit, which is phosphorylated by both these enzymes . Limited tryptic digestion and subsequent cyanogen bromide treatment of rat liver eIF-2 phosphorylated by HCR yielded one major phosphopeptide . This peptide had the sequence Ile-Leu-Leu-Ser-Glu-Leu-Ser(P)-Arg-Arg . The same major phosphopeptide was obtained from rabbit reticulocyte eIF-2 phosphorylated by HCR or dsI as judged by its behaviour on two-dimensional mapping and reverse-phase chromatography . In all cases the phosphorylated residue was found to be serine-7, and not serine-4, of the above sequence as determined from sequence analysis and by subdigestion of the peptide with Staphylococcus aureus V8 proteinase.

J Biol Chem, 1987 Jul 15, 262(20), 9676 - 80
Identification of cysteine 656 as the amino acid of hepatoma tissue culture cell glucocorticoid receptors that is covalently labeled by dexamethasone 21-mesylate; Simons SS Jr et al.; Recent results using proteases suggest that dexamethasone 21-mesylate (Dex-Mes) labeling of the rat hepatoma tissue culture (HTC) cell glucocorticoid receptor occurs at one or a few closely grouped cysteine residues (Simons, S.S., Jr . (1987) J . Biol . Chem . 262, 9669-9675) . In this study, a more direct approach was used both to establish that only one cysteine is labeled by {3H}Dex-Mes and to identify the amino acid sequence containing this labeled cysteine . Various analytical procedures did not provide the purification of the extremely hydrophobic Staphylococcus aureus V8 protease digestion fragment that is required for unique amino acid sequencing data . Therefore, Edman degradation was performed on the limit protease digest mixtures which appeared to contain only one 3H-labeled peptide . These degradation experiments revealed the number of amino acid residues between the NH2 terminus of each peptide and the {3H}Dex-Mes-labeled cysteine . A comparison of these amino acid spacings with the published amino acid sequence of the HTC cell glucocorticoid receptor (Miesfeld, R., Rusconi, S., Godowski, P . J., Maler, B . A., Okret, S., Wikstom, A-C., Gustafsson, J-A., and Yamamoto, K . R . (1986) Cell 46, 389-399) indicated that the one cysteine labeled by {3H}Dex-Mes is Cys-656 . Further analysis of the receptor sequence for the presence of the observed grouping of proteolytic cleavage sites, but without any preconditions as to which amino acid was labeled, gave Asp-122 and Cys-656 as the only two possibilities . Potential labeling of Asp-122 could be eliminated on the basis of immunological and genetic evidence . We, therefore, conclude that the single Dex-Mes-labeled site of the HTC cell glucocorticoid receptor has been identified as Cys-656 . Since several lines of evidence indicate that {3H}Dex-Mes labeling of the receptor occurs in the steroid binding site, Cys-656 is the first amino acid which can be directly associated with a particular property of the glucocorticoid receptor.

J Biol Chem, 1987 Jul 15, 262(20), 9669 - 75
Selective covalent labeling of cysteines in bovine serum albumin and in hepatoma tissue culture cell glucocorticoid receptors by dexamethasone 21-mesylate; Simons SS Jr; The specificity of protein labeling by an affinity label of glucocorticoid receptors, dexamethasone 21-mesylate (Dex-Mes), was investigated using bovine serum albumin (BSA) as a model . During the early stages of {3H}Dex-Mes labeling at pH 8.8, approximately 90% of the covalent bond formation occurred at the one non-oxidized cysteine (Cys-34) of BSA . The nonspecific labeling was equally distributed over the rest of the BSA molecule . {3H}Dex-Mes labeling of Cys-34 was totally, and specifically inhibited by nearly stoichiometric amounts of the thiol-specific reagent methyl methanethiolsulfonate (MMTS) . Thus both Dex-Mes and MMTS appear to react very selectively with thiols under our conditions . In reactions with hepatoma tissue culture (HTC) cell glucocorticoid receptors, MMTS was equally efficient in preventing {3H}dexamethasone binding to receptors and {3H}Dex-Mes labeling of the 98-kDa receptor protein . These results indicate that Dex-Mes labeling of the glucocorticoid receptor involves covalent reaction with at least one cysteine in the steroid binding site of the receptor . Small (approximately 1600-dalton) fragments of the {3H}Dex-Mes-labeled 98-kDa receptor were generated by limit proteolysis with trypsin, chymotrypsin, and Staphylococcus aureus V8 protease under denaturing conditions . Data from these fragments on 15% sodium dodecyl sulfate-polyacrylamide gels were consistent with all of the covalent {3H} Dex-Mes being located on one or a few cysteines in one approximately 15-residue stretch of the receptor . Further studies revealed no differences in the limit protease digestion patterns of activated and unactivated {3H}Dex-Mes-labeled receptors with trypsin, chymotrypsin, or V8 protease under denaturing conditions . These data suggest that activation does not cause any major covalent modifications of the amino acids immediately surrounding the affinity-labeled cysteine(s) of the steroid binding site.

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