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 an