Staphylococcus aureus

Chemotherapy, 1991, 37(4), 251 - 5
The effects of ions on antibacterial activity of ofloxacin and ceftriaxone; Gurdal H et al.; MIC and MBC values of ofloxacin and ceftriaxone were investigated against Staphylococcus aureus in MHB and MHB containing additional Mg2+, Al3+, Fe3+, Ca2+, Zn2+, Cu2+ . The addition of Mg2+, Al3+, Fe3+ increased the MIC and MBC of ofloxacin and the MBC of ceftriaxone . However, the addition of these cations did not change the MIC of ceftriaxone . Our findings suggest that these interactions might be due to the formation of chelates between metal ions and antibiotics . These results also indicate that some cations may have an important role in the antibacterial activity of antibiotics.

Acta Cardiol, 1991, 46(5), 589 - 92
Illustration by transesophageal echocardiography of rapid and important pannus formation during infective endocarditis of a prosthetic valve; Van Camp G et al.; A 38-year-old man with a Starr-Edwards mitral prosthetic valve presented with a Staphylococcus aureus septicemia . Twenty-four hours later, transthoracic echocardiography did not show obvious vegetations but Doppler examination of the prosthetic valve demonstrated a prolonged half pressure time and an elevated peak transmitral velocity . Seventy-two hours after the first septic event transesophageal echocardiography revealed a large annular pannus floating in the left atrium in systole and protruding in the prosthetic cage during diastole . This case report emphasizes the importance of transesophageal echocardiography in septic patients with prosthetic valves and underlines the possibility of extremely rapid pannus formation in these patients.

Acta Vet Hung, 1991, 39(3-4), 187 - 95
The involvement of polymorphonuclear leukocytes in the pathogenesis of bronchopneumonia in calves . II . Granulocyte-induced changes in erythrocyte membrane phospholipid topology; Ledwozyw A et al.; Phospholipid topology in erythrocyte membranes of bronchopneumonic calves and changes in phospholipid asymmetry induced by incubation of erythrocytes with neutrophils of diseased calves were examined . Using aminophospholipid tracing by non-penetrating probe, trinitrobenzenesulfonic acid, and phospholipid hydrolysis by specific phospholipase A2 from Naja naja venom and sphingomyelinase from Staphylococcus aureus it was stated that in erythrocytes from diseased calves phosphatidylserine and, to some extent, phosphatidylethanolamine are externalized on the outer leaflet of the membrane . Similar results were obtained for erythrocytes of healthy calves after incubation with granulocytes of bronchopneumonic animals . It is suggested that granulocytes of bronchopneumonic calves cause perturbations in erythrocyte membrane phospholipid organization.

Med Dosw Mikrobiol, 1991, 43(1-2), 11 - 7
{Humoral response to Staphylococcus aureus antigens evaluated by the western blotting method}; Tyski S et al.; Cellular antigens extracted from the cells of four Staphylococcus aureus strains from different kinds of infections (sepsis, osteomyelitis, furunculosis) were analysed by the western blotting technique . Antibiotic sensitivity pattern of the strains was compared . One isolate was found to be MRSA strain . Sera samples from patients of whom strains were isolated and four sera from blood donors (as a control) were used in the investigation . IgG levels for purified staphylococcal antigens (lipase, alpha-toxin and teichoic acid) were estimated . Interaction between extracted bacterial antigens and serum antibodies of IgG class were analysed in homologous and heterologous systems . The most strong immunological reaction of the investigated sera with staphylococcal antigens was observed in the case of homologous system . Serum from sepsis patient was found to be the most reactive serum with all staphylococcal antigens mixtures.

Med Dosw Mikrobiol, 1991, 43(1-2), 1 - 10
{Preparation of Staphylococcus aureus antigens for evaluation of their immunological reactivity with the human sera by the western blot method}; Tyski S et al.; Extracellular antigens as well as cell wall extracts of 4 S . aureus strains isolated from different kinds of infection were analysed by Western-Blott technique . Materials obtained in two systems of bacteria cultivation (with and without aeration) were compared . Four systems of PAGE (native conditions, with 8.0 M urea, with SDS and SDS after previous reduction of the material with 2-mercaptoethanol) were compared in order to get the best differentiation of proteins and antigens . Immunological reactivity of the antigens mixture with two human sera: highly positive (with three S . aureus antigens in ELISA) from patient with staphylococcal sepsis and negative (from blood donor) were analysed . The best results were obtained after reduction of the cell wall extracted material in SDS-PAGE . The different protein patterns depending on the strain and the method of bacteria cultivation were observed . The standardisation of Western-Blott technique was performed, including titration of the sera to get the best differentiation of the antigens . The difference in immunological reactivity of the positive and negative sera with staphylococcal antigens mixture showed rather quantitative than qualitative character.

Exp Pathol, 1991, 43(1-2), 63 - 5
Pulmonary histiocytosis X cells phagocytize Staphylococcus aureus in vitro; Hurter T et al.; The broncho-alveolar lavage from an 18-year-old girl with proven histiocytosis X of the lung has been cocultivated with Staphylococcus aureus . Electron microscopy revealed that the bacteria adhere to the membrane of HX cells and become ingested afterwards.

Cytometry, 1991, 12(7), 687 - 93
Simultaneous flow cytometric method to measure phagocytosis and oxidative products by neutrophils; Perticarari S et al.; We developed a rapid and sensitive two-color flow cytometric method which allows the simultaneous quantification of both the phagocytosis rate and the oxidative burst activation of polymorphonuclear leukocytes (PMNLs) . The oxidation of hydroethidine (HE) to ethidium bromide (EB) was performed by the oxidative neutrophil products within the cells during the respiratory burst, which was stimulated by phagocytized fluorescein-labeled Staphylococcus aureus . By means of flow cytometry we measured red EB fluorescence emission together with green fluorescence, which was emitted by the ingested fluoresceinated bacteria . The fluorescence intensity was proportional to the number of bacteria ingested . Adherent bacteria were distinguished from the ingested ones . This two-color cellular staining permits measurement of two different functions of neutrophils in one step . This method could be of interest for the determination of the interactions between neutrophils and bacteria and for the investigations on infectious diseases in chronic granulomatous disease patients.

Scand J Urol Nephrol, 1991, 25(4), 307 - 10
Prognosis in septicemia complicated by acute renal failure requiring dialysis; Frost L et al.; During a 12-year period 419 patients were admitted because of acute renal failure requiring dialysis . Fifty (12%) had septicemia verified by blood culture . In a retrospective study age, sex, focus of infection, blood culture results, kidney function, mode of dialysis treatment, numbers and durations of complicating organ failures, presence of gastrointestinal bleeding, and secondary complicating events of septicemia were recorded for the purpose of establishing a prognostic index based on clinical criteria . Respiratory failure was present in 34 patients, circulatory failure in 31 patients, failure of coagulation system in 25 patients, and hepatic failure in 10 patients . Overall mortality was 46% . Highest death-rates were found during the first days of dialysis . In patients with multiple organ failures, in elderly and in patients suffering from staphylococcus aureus septicemia, a non-significant trend towards higher mortality was found . The mode of dialysis treatment did not influence patient survival . Our intention of establishing a prognostic index based on bedside clinical criteria has not been fulfilled . Even though mortality-rate increases in patients with acute renal failure complicated by failure of one or more vital organs, survival-rate in patients with four or more organ failures was 30%.

Int Arch Allergy Appl Immunol, 1991, 96(2), 102 - 6
Tumor necrosis factor production by human granulocytes; Mandi Y et al.; Human polymorphonuclear leukocytes kill WEHI 164 cells in an 18-hour 51Cr release assay . Antibody to human tumor necrosis factor (TNF) blocks the lysis of targets mediated by human granulocytes . Resting granulocytes produce an undetectable amount of TNF, if any . Granulocytes stimulated with Staphylococcus aureus release 250-500 U/ml TNF alpha . The specificity of the released TNF in the WEHI 164 cytotoxicity assay was confirmed by using neutralizing anti-TNF alpha monoclonal antibodies . The thymidine uptake of endothelial cells was inhibited by granulocyte-derived TNF . The identity of TNF alpha was further confirmed by molecular weight determination, by gel filtration on Sephacryl S-200, with a result of approximately 44,000 . Besides their antimicrobial capacity, therefore, granulocytes may contribute to tumor rejection, inflammation and septic infections by releasing TNF.

Scand J Infect Dis, 1991, 23(5), 617 - 23
Efficacy of cloxacillin prophylaxis in craniotomy: a one year follow-up study; Van Ek B et al.; A previous prospective double-blind placebo-controlled study showed that cloxacillin prophylaxis significantly reduces the rate of infection after craniotomy . The purpose of the present follow-up study was to find out whether the data on routine cloxacillin prophylaxis would confirm the decreased infection rate after craniotomy . During this 1-year study, 8 infections occurred after 201 operations (4.0%) in 175 patients . No prophylaxis was given during 17 operations in 15 patients; 4 infections occurred in this group . The most frequently isolated microorganisms were Staphylococcus aureus, S . epidermidis, and Propionibacterium acnes, all of which were sensitive to cloxacillin . We conclude that cloxacillin is beneficial as prophylaxis in patients undergoing craniotomy.

Scand J Infect Dis, 1991, 23(5), 613 - 6
Neonatal omphalitis is still common in eastern Turkey; Guvenc H et al.; 88 newborns with omphalitis diagnosed at a University Hospital in Eastern Turkey from January 1988 to December 1990 were reviewed . The yearly incidence was 7.7% in inpatient newborns . Risk factors including septic delivery (especially home delivery), prematurity, and being small for gestational age appeared to be important . Clinical manifestations, white blood cell and absolute neutrophil counts were non-specific and non-pathognomonic . Bacteriologic analysis revealed that Gram-positive bacteria (68%) predominated over Gram-negative isolates (60%) . Staphylococcus aureus and Escherichia coli were the most frequent microorganisms . Mortality rate was 15%; all deaths occurred in patients presenting with systemic symptoms . The following poor prognostic factors were identified: septic delivery (particularly at home), prematurity, being small for gestational age and the development of systemic symptoms.

Br J Neurosurg, 1991, 5(5), 515 - 7
Cysto-peritoneal shunt infection with Trichosporon beigelii; Ashpole RD et al.; Infection is a frequent problem affecting cysto-peritoneal shunts, the usual causative organisms being Staphylococcus epidermidis and Staphylococcus aureus . Fungi are rarely isolated from such infections . We present the first report, as far as the authors are aware, of a case of Trichosporon beigelii shunt infection that responded to shunt removal and antifungal therapy.

Chemotherapy, 1991, 37(6), 420 - 5
Postantibiotic effect of ciprofloxacin compared with that of five other quinolones; Minguez F et al.; The antimicrobial activity (minimal inhibitory concentration, MIC, and killing kinetics) and postantibiotic effect (PAE) of different concentrations (MIC and 6 mg/1) of ciprofloxacin, norfloxacin, ofloxacin, pefloxacin, fleroxacin and lomefloxacin on pure cultures of Staphylococcus aureus and Escherichia coli were compared in vitro . The MIC, killing kinetics and PAE were determined by standard methods . Ciprofloxacin displayed the lowest MICs, while the highest were those of norfloxacin against S . aureus and lomefloxacin against E . coli . The killing curves showed ciprofloxacin to be the most and norfloxacin the least active . The bactericidal power was dependent on the concentration . At MIC, the fluoroquinolones, with the exception of norfloxacin, induced PAEs of 1-2 h . The effect was, in all cases, greater against E . coli . When assayed at 6 mg/l the PAEs were increased to 2-5 h, the best results being obtained by ciprofloxacin followed by ofloxacin . Norfloxacin produced no PAE on S . aureus and scarcely reached 1.3 h against E . coli . There was a close relationship between bactericidal power and PAE.

Microbios, 1991, 67(274), 53 - 63
Effect of Cd2+ on phosphate uptake by cadmium-resistant and cadmium-sensitive Staphylococcus aureus; Tynecka Z et al.; The effect of Cd2+ on phosphate (Pi) uptake was investigated in the growing cells of Cd(2+)-resistant Staphylococcus aureus 1781OR and Cd(2+)-sensitive S . aureus 17810S . Inhibitor and ionophore studies showed that 32Pi uptake in the two strains occurred via the Pi porter down pH gradient (delta pH) generated by the respiratory chain . Cd2+ inhibited 32Pi uptake in the cadmium-sensitive strain 1781OS at all concentrations used (10 microM-1 mM) . In strain 1781OR, possessing the plasmid-coded Cd2+ efflux system, 10-100 microM Cd2+ did not inhibit 32Pi uptake . Even at 1 mM Cd2+, inhibition of 32Pi uptake in strain 1781OR was reversed when the external Cd2+ was chelated with cysteine and activity of Cd2+ efflux system was restored . Cd2+ efflux induced by cysteine was energized either by membrane potential (delta psi) or by delta pH, which indicated that electrochemical gradient of protons (delta mu H+) was required for this efflux.

Comp Biochem Physiol B, 1991, 100(1), 77 - 81
A comparative study of the structure of egg-white riboflavin binding protein from the domestic fowl and Japanese quail; Walker M et al.; 1 . The riboflavin binding proteins from domestic fowl and Japanese quail have been isolated and their structures compared by circular dichroism, fluorescence and peptide mapping . 2 . The two proteins have similar secondary structures, but differ in their tertiary structures as reflected in the environments of aromatic amino acid side chains . 3 . Differences in amino acid sequence between the proteins are indicated by the digestion patterns obtained with thermolysin, chymotrypsin and V8 proteinase from Staphylococcus aureus . Both proteins are resistant to digestion by trypsin.

Eur J Cardiothorac Surg, 1991, 5(10), 515 - 8
Antibiotic prophylaxis in non-cardiac thoracic surgery: cefazolin versus placebo; Aznar R et al.; A total of 127 patients (57 given placebo and 70 given a single preoperative dose of 1 g cefazolin) undergoing thoracic surgery were included in a randomized double-blind trial . The two groups were similar in regard to mean age, sex ratio, in-hospital stay before surgery, underlying disease, risk factors, type of surgery, mean duration of surgical procedure, and mean duration of chest tube drainage . The relative risk of wound infection of the patients from the placebo group was 3.27 (range 1.5-11.5; 95% confidence interval) . Cefazolin significantly reduced (p less than 0.01) the wound infection rate--1 case (1.5%) in the cefazolin group versus 8 cases (14%) in the placebo group--but not the incidence of postoperative pleural empyema--5 (7%) versus 8 cases (14%)--or nosocomial pneumonia--3 (4%) versus 5 cases (9%) . Cultures were made from 3 out of 9 wound infections and Staphylococcus aureus or S . epidermidis was isolated in all 3 . In addition, cultures were made from 6 out of 13 pleural cavity infections and S . aureus (1 case) or other microorganisms (5 cases) were isolated in all 6 . Mortality was similar in both groups and all deaths unrelated to the infections . No adverse side effects of the drug were encountered . In conclusion, a single preoperative dose of 1 g cefazolin proved to be effective for reducing the wound infection rate in non-cardiac thoracic surgery.

Perit Dial Int, 1991, 11(4), 333 - 40
Exit-site infection in continuous ambulatory peritoneal dialysis: a review; Luzar MA; Although the ability of CAPD to successfully treat end-stage renal disease is now well established, exit-site infection (ESI) remains a serious cause of morbidity . The objective of this article is to review recent advancements relating to ESI pathogenesis and its reduction . Current definitions of ESI are reviewed, as are comparative studies of etiology . Emphasis is placed on the literature identifying Staphylococcus aureus as the primary cause of ESI in CAPD . The article reviews reported rates of ESI and discusses reasons for variations of this complication's reported frequency . The selection of catheters available in CAPD are discussed, as are studies demonstrating the threat of S . aureus to catheter survival . The pathogenesis of exit-site infection related to S . aureus nasal carriage in CAPD is reviewed in light of recent findings indicating the pre-CAPD nasal carrier as the patient at risk for subsequent ESI . Postoperative and long-term care of the catheter patient are reviewed for various literature protocols . Treatment recommendations for choice of agents are discussed . Future research should include a better understanding of the morphology of the CAPD catheter exit-site in humans and the healing process . ESI epidemiological studies should be encouraged in tandem with well-designed, controlled studies on the value of prophylactic treatment.

Int Orthop, 1991, 15(3), 205 - 8
Complications of suppurative arthritis and osteomyelitis in children; Porat S et al.; Three children with suppurative arthritis and osteomyelitis are described to emphasise that delayed or incorrect diagnosis may lead to serious cardiopulmonary complications . In two patients, bilateral bronchopneumonia developed with pneumatocoeles, pneumothorax and empyema . The other had cardiac failure from septic pericarditis . In one case, disarticulation of the knee was needed as a life-saving measure, and the other leg developed an infected pseudarthrosis of the tibia . The causative organism in each case was staphylococcus aureus and no evidence of immunodeficiency was demonstrated.

Dermatologica, 1991, 183(2), 123 - 8
Lack of correlation between in vitro immunological alterations and the development of scleroderma-like skin lesions in toxic oil syndrome patients; Gutierrez C et al.; We examined whether immunological disturbances could influence the development of scleroderma-like skin lesions in patients affected by the Spanish toxic oil syndrome (TOS) . To this end, peripheral blood mononuclear cells (PBMC) were collected from 13 chronic patients and 8 control subjects . All patients had suffered a toxic-induced severe neuromyopathy, and 6, in addition, had developed sclerodermoid skin manifestations . The phenotypic profile and the concentrations of interleukin-2 (IL-2) and of molecules with B cell differentiation factor IgG activity (BCDF-IgG) in supernatants of phytohemagglutinin-stimulated lymphocytes were analyzed both in patients and in normal controls . Molecules with BCDF-IgG activity were found increased in supernatants of mitogen-stimulated lymphocytes from TOS patients . Concentrations of IgG secreted by staphylococcus aureus-SAC-B blasts in the presence of TOS PBMC supernatant was 88 +/- 32.62 ng/ml (mean +/- 1 SD) versus 53 +/- 5.34 ng/ml in the presence of control supernatant (p less than 0.01) . Levels of BCDF-IgG activity in TOS PBMC supernatants positively correlated with IgG serum levels (r = 0.69, p less than 0.01) . The phenotypic profile of lymphocyte populations and the production of IL-2 were not altered in TOS subjects . No statistically significant differences were observed in the lymphocyte distribution nor in the IL-2 and BCDF-IgG production when comparing patients with or without scleroderma-like skin lesions . The results indicate that there was a dysfunction of the immune response in TOS subjects, which, however, was not sufficient for the development of the sclerodermoid lesions.

Eur Biophys J, 1991, 20(4), 229 - 40
Ion channels formed by amphipathic helical peptides . A molecular modelling study; Sansom MS et al.; Channel forming peptides (CFPs) are amphipathic peptides, of length ca . 20 residues, which adopt an alpha-helical conformation in the presence of lipid bilayers and form ion channels with electrophysiological properties comparable to those of ion channel proteins . We have modelled CFP channels as bundles of parallel trans-bilayer helices surrounding a central ion-permeable pore . Ion-channel interactions have been explored via accessible surface area calculations, and via evaluation of changes in van der Waals and electrostatic energies as a K+ ion is translated along the length of the pore . Two CFPs have been modelled: (a) zervamicin-A1-16, a synthetic apolar peptaibol related to alamethicin, and (b) delta-toxin from Staphylococcus aureus . Both of these CFPs have previously been shown to form ion channels in planar lipid bilayers, and have been shown to have predominantly helical conformations . Zervamicin-A1-16 channels were modelled as bundles of 4 to 8 parallel helices . Two related helix bundle geometries were explored . K(+)-channel interactions have been shown to involve exposed backbone carbonyl oxygen atoms . delta-Toxin channels were modelled as bundles of 6 parallel helices . Residues Q3, D11 and D18 generate favourable K(+)-channel interactions . Rotation of W15 about its C beta-C gamma bond has been shown to be capable of occluding the central pore, and is discussed as a possible model for sidechain conformational changes in relation to ion channel gating.

Autoimmunity, 1991, 10(2), 153 - 63
Structure-function studies of S-antigen: use of proteases to reveal a dominant uveitogenic site; Dua HS et al.; Retinal S-antigen induced experimental autoimmune uveitis (EAU) is a severe, predominantly T-cell mediated inflammatory disease of the uveal tract and retina of the eye . Pretreatment of LEW rats with the monoclonal antibody, MAbS2.4.C5, which defines an epitope in S-antigen, has been shown to effectively inhibit the subsequent induction of EAU with S-antigen . Using synthetic peptides and cyanogen bromide fragments of S-antigen we found the binding site of MAbS2.4.C5 to be located at the carboxy terminus of the molecule corresponding to amino acid positions 375 to 380 . Limited Staphylococcus aureus V8 protease digestion yielded several polypeptide fragments including one large 43 kD fragment which retained antibody binding to a variety of both polyclonal and monoclonal antibodies which identify epitopes that span the length of the S-antigen . This treatment, however, completely destroys the MAbS2.4.C5 binding site and dramatically reduces uveitopathogenicity . Limited trypsin and papain digestion, on the other hand, had little effect on pathogenicity or on MAbS2.4.C5 binding to S-antigen or its peptide fragments . These results indicate that the carboxy-terminus of S-antigen plays a predominant role in the pathogenesis of EAU.

Lab Delo, 1991, (5), 53 - 4
{The use of semi-starvation nutrient media for phage typing of Staphylococcus aureus}; Filichkin SE et al.; A total of 161 Staphylococcus aureus strains grown in Hottinger's broth and semi-starvation medium (1% peptone water with 1% glycerol) were phage-typed . 107 (66.4 +/- 3.1%) of the 161 cultures grown in the semi-starvation medium could be typed and only 77 (48.1 +/- 3.63%) of the 161 grown in Hottinger's broth, which fact speaks in favor of semi-starvation nutrient media employment for phage-typing of S . aureus.

Ann Trop Paediatr, 1991, 11(1), 41 - 5
A survey of nasal carriage of Staphylococcus aureus in a neonatal ward in Ile-Ife, Nigeria; Ako-Nai AK et al.; The nasal carriage rate of Staphylococcus aureus among maternal-infant pairs was 18% compared with 39% among hospital staff in Ile-Ife, Nigeria during a 12-week survey . Of the newborns, 46% tested positive compared with 26% of their mothers . The S . aureus phage types recovered were predominantly of the group III type (38%); however, 28% of the strains isolated were non-typable . All the S . aureus strains were resistant to penicillin, 84% to tetracycline, and 35 and 24% were resistant to streptomycin and chloramphenicol, respectively . Altogether 19% of the strains tested were resistant to methicillin.

Acta Otolaryngol, 1991, 111(2), 420 - 7
Antigen-specific plaques formation of cultured mononuclear cells in head and neck cancer; Wustrow TP; Antigen-specific antibody production in vitro from peripheral blood mononuclear cells was studied for the assessment of the immune competence of patients with head and neck cancer . Optimal culture conditions were studied using inactivated Staphylococcus aureus and a saturation with IL-1 during B cell activation and pooled human AB serum on day 2 . After passage of the mononuclear cells through sephadex G-10 columns, a significant increase in the antigen-specific antibody production was observed . In healthy donors a significant reduction of the antigen-specific antibody production according to the abuse of alcohol and/or cigarette smoking was detectable . Interestingly, high alcohol consumption resulted in a more pronounced decrease of the antigen-specific antibody production in vitro than excessive cigarette smoking . Patients with squamous cell carcinoma of the head and neck who are considered to be most immunodeficient did not show any antigen-specific antibody production in vitro upon activation with sheep red blood cells in the presence of Interleukin-1 (IL-1) . After filtration of mononuclear cells from peripheral bloodover sephadex G-10 beads, two thirds of the patients studied became stimulable . This increase in the antigen-specific antibody production in vitro was significant, though not as dramatic as in the age- and sex-matched control groups . Interestingly, the antigen-specific antibody production raised almost to the same level as that measured in healthy donors with high alcohol abuse and cigarette consumption.

Allergy, 1991 Jan, 46(1), 45 - 51
Bacteria-induced histamine release from human bronchoalveolar cells and blood leukocytes; Clementsen P et al.; Histamine release induced by Staphylococcus aureus was examined in cells obtained by bronchoalveolar lavage (BAL) in non-atopic individuals . Approximately half of the individuals responded with mediator release to the bacterium, and the release was found to be time- and concentration dependent . No difference was found between the patients who responded and those who did not respond in regard to age, sex, smoker/non-smoker, % recovery of BAL-fluid, total cell count, differential cell counts, histamine content per mast cell, or diagnoses . Also stimulation of the BAL-cells with the calcium-ionophore A23187 resulted in histamine release . S . aureus-induced histamine release from basophils was examined in leukocyte suspensions obtained from the same individuals, and in all experiments release was found . The dose-response curves were similar to those obtained with BAL cells . The bacteria-induced mediator release from superficially lying cells in the airways epithelium might be of importance for the precipitation or exacerbation of bronchial asthma in respiratory tract infections.

J Membr Biol, 1991 Jan, 119(1), 53 - 64
Modification of lysine residues of Staphylococcus aureus alpha-toxin: effects on its channel-forming properties; Cescatti L et al.; Staphylococcus aureus alpha-toxin opens an ion channel in planar phospholipid bilayers, which is selective for anions over cations, supposedly because of the presence of positively charged groups along the ion pathway . To remove some positive charges of this protein toxin, we chemically modified part of its lysine residues either with diethylpyrocarbonate, followed by histidine regeneration with hydroxylamine, or with trinitrobenzenesulfonic acid . The extent of chemical modification can be followed accurately by native polyacrylamide gel electrophoresis and isoelectric focusing . Ethoxyformilation of two to three lysine residues per toxin monomer does not impair hemolysis of rabbit red blood cells nor formation of pores in model membranes . It reduces the conductance and the anion selectivity of the channel and changes the shape of its current-voltage characteristic . This indicates that positively charged lysine residues are actually important in determining the electrical properties of the pore . Ethoxyformilation of channels preassembled in planar bilayers produces the same changes as modification of toxin monomers before channel formation . Furthermore, it can be performed by adding diethylpyrocarbonate on either side of the bilayer . This suggests that the lysine residues relevant for the electrical properties of the pore are located inside its lumen where they can be reached by diethylpyrocarbonate diffusing from either entrance of the channel.

J Antibiot (Tokyo), 1991 Jan, 44(1), 86 - 92
Synthesis and activity of butirosin derivatives with 5''-amidino and 5''-guanidino substituents; Narita Y et al.; The preparation and antibacterial activity of the 5''-guanidino (6) and 5''-amidino (7) derivatives of 4'-deoxybutirosin A (1) as well as the 5''-guanidino derivative (8) of butirosin A are described . The key intermediates, tetra-N-benzyloxycarbonyl-5''-azido derivatives were selectively reduced with NiCl2-NaBH4 to give the corresponding 5'-amino derivatives . Subsequent guanidination or amidination followed by deblocking afforded the final compounds 6, 7 and 8 . The 5''-guanidino derivatives (6 and 8) were more active against Gram-positive and Gram-negative bacteria than the corresponding 5''-hydroxy derivatives (1 and butirosin A) . Compound 6 was also active against a variety of methicillin-resistant Staphylococcus aureus (MRSA).

Adv Perit Dial, 1991, 7, 81 - 5
Subcutaneous versus intraperitoneal insulin in the management of diabetics on CAPD: a review; Tzamaloukas AH et al.; Intraperitoneal and subcutaneous routes of administration for diabetics on CAPD were compared . The comparison included: (1) Control of blood glucose concentration: both methods can provide satisfactory glycemic control for most patients . Changing the method of insulin administration is warranted when one method fails . (2) Effect on plasma insulin levels: intraperitoneal administration can produce a plasma insulin profile similar to the normal profile . This is unusual with subcutaneous administration . Consequences of hyperinsulinemia (hyperlipidemia, hypertension) seem, however, to be similar between the two methods of insulin administration . (3) Effect on peritoneal permeability: permeability characteristics are maintained unchanged, usually, with either method after long-term CAPD . However, insulin is mitogenic in vitro . Theoretically, intraperitoneal insulin could lead to peritoneal fibrosis . (4) Effect on infectious complications of CAPD: a difference in the rate of peritonitis or overall PD catheter-related infections has not been convincingly demonstrated between the two methods of insulin administration . Exit site and tunnel infections with staphylococcus aureus may be more frequent in diabetics receiving insulin subcutaneously . (5) Effect on hepatic structure and function: subcapsular hepatic steatosis was described in diabetics receiving insulin intraperitoneally . The clinical significance of this finding remains to be demonstrated . We conclude that both methods can be applied for insulin administration in diabetics on CAPD . The intraperitoneal method should be tried first in most instances . Prospective studies comparing the two methods are needed.

Adv Perit Dial, 1991, 7, 196 - 203
Peritonitis rates for CAPD patients using the SCD 210 (Inpersol sterile connecting device): a Canadian survey; Lafleur D et al.; A retrospective study was conducted in 20 Canadian hospitals to assess peritonitis rates of CAPD patients utilizing the SCD 210 patient assist device during a 30 month study period . A total of 175 patients having a cumulative SCD experience of 1,494 patient months were included in the survey . Sixty-eight patients experienced 116 episodes of peritonitis, i.e., one episode every 12.9 patient months (pt mo) . Patients that had used the SCD for their entire CAPD experience had a significantly lower peritonitis rate (1/15.2 pt mo) compared to patients who had used other CAPD systems (1/10.1 pt mo) . Peritonitis rates for diabetic study patients or patients with impaired vision and/or dexterity were not significantly greater than non-diabetic or non-impaired study patients . Staphylococcus epidermidis and Staphylococcus aureus were the most frequent causative microorganisms, accounting for 27.7% and 16.0% of peritonitis episodes caused by single organisms, respectively . The proportion of peritonitis caused by skin commensals was consistent across all study patients, regardless of impairment of vision and/or dexterity . The study results demonstrate the successful application of this patient assist device in a CAPD population that consisted of a large proportion of high risk patients.

Adv Perit Dial, 1991, 7, 125 - 8
Active intervention dramatically reduces CAPD-associated infection; Dryden MS et al.; In 1987 a preventive programme was initiated to address the problem of high hospital and community-acquired CAPD infection . It concentrated on reducing Staphylococcus aureus carriage, improving aseptic operative technique, intensive training for nursing staff and patients in stringent aseptic care of the exit site, and avoidance of contact of the exit site with unsterile water . This programme was associated with an overall 10-fold reduction in exit site infection, a 2-fold reduction in peritonitis, and a 4.5-fold reduction in catheter loss from infection . These reductions have been sustained . Preventing infection in CAPD patients requires persistence and commitment but improves the patient's quality of life and reduces the cost of treatment.

Adv Perit Dial, 1991, 7, 117 - 9
Grading of exit sites cannot predict peritonitis in patients on CAPD; Ahlmen J et al.; Grading of exit sites was performed at 265 routine outpatient visits of 28 patients starting CAPD between January 1988 and February 1990, with a total observation time of 398.5 months to December 1990 . Six patients had no peritonitis episode . The remaining 22 patients suffered from 43 peritonitis episodes . Fifty-eight per cent of these were caused by Staphylococcus epidermidis . Eighty-six per cent of the examinations of exit sites of patients with peritonitis episodes showed 0 to 1 point on an arbitrary grading scale of up to 9 points . Staphylococcus aureus was found after bacterial culture from exit site smears in 3 cases . All were adequately treated . None of these cases showed any peritonitis episodes with this bacteria . No relationship could be found between the grading of the exit site at routine outpatient clinic visits and the appearance of a peritonitis episode . Grading of exit sites is of clinical importance but cannot predict the appearance of peritonitis.

Eur J Surg, 1991 Jan, 157(1), 67 - 8
Intraperitoneal rupture of an infected urachal cyst in an infant . Case report; Chen TF et al.; Peritonitis due to intraperitoneal rupture of an infected urachal cyst is a life-threatening condition, not previously reported in an infant . We report this condition in an 8-month-old infant . The child underwent complete excision of the urachal remnant and made an uneventful recovery; bacterial culture grew Staphylococcus aureus . Subsequent investigations did not reveal any other renal tract abnormalities.

J Hosp Infect, 1991 Jan, 17(1), 25 - 33
Epidemiological study of Staphylococcus aureus resistance to new quinolones in a university hospital; Aboukasm AG et al.; During a 14-month period, from December 1984 to February 1986, 630 Staphylococcus aureus isolates were identified at Broussais Hospital . Thirty-eight isolates (6%), from 35 patients, were found to be pefloxacin-resistant S . aureus (PRSA) with minimal inhibitory concentrations greater than or equal to 8 mg l-1 . PRSA isolates were tested for susceptibility to 35 antibiotics, including nine quinolones, and heavy metal ions . Phage-type was determined . Out of the 38 PRSA isolates, 35 (92%) were methicillin- and multiply-resistant; however, all PRSA isolates were sensitive to vancomycin and coumermycin . Fifteen isolates (39%) had similar phage-type and identical antibiotic susceptibility pattern with high level resistance to pefloxacin (MICs equal to 64 mg l-1); they were isolated from the same surgical unit . The 23 remaining PRSA isolates differed by their phage and susceptibility patterns . Pefloxacin MICs ranged from 8 to 512 mg l-1 with a bimodal distribution; cross-resistance was observed with the eight other quinolones tested . Only nine PRSA isolates (24%), including four 'epidemic' isolates, were obtained from patients who had been treated with quinolones . From these data there is apparently no direct relationship between quinolone administration and selection of PRSA in infected patients.

Chemotherapy, 1991, 37(5), 335 - 42
Effect of ampicillin/sulbactam on human polymorphonuclear leukocyte function; Pascual A et al.; The effect of ampicillin and sulbactam on the interaction in vitro of human polymorphonuclear leukocytes (PMN) with Staphylococcus aureus was examined . The exposure of a non-penicillinase-producing S . aureus strain to one fourth the MIC of ampicillin but not of sulbactam significantly increased the uptake of bacteria by human PMN . This effect was also observed when bacteria were exposed to the MIC0.25 of different combinations of ampicillin and sulbactam (2/1, 1/1, 1/4, 1/8 and 1/32) . These effects were not observed when a penicillinase-producing strain was used . The production of superoxide and hydrogen peroxide radicals by human PMN was not affected by the presence of these antimicrobials . Ampicillin and sulbactam, neither alone nor in combination, showed intracellular activity against S . aureus within human PMN.

Int J Immunopharmacol, 1991, 13(4), 419 - 28
Effect of anti-leprosy drugs on superoxide anion production by rat peritoneal macrophage with special reference to light exposed clofazimine; Sahu A et al.; The present study describes the in vitro effect of anti-leprosy drugs on superoxide anion (O2-) production by rat resident peritoneal macrophages . Of the three drugs tested i.e . clofazimine, rifampicin and dapsone, the first was most effective in increasing O2- production in a dose dependent manner, while rifampicin had some stimulatory effect and dapsone exhibited minimal action . Furthermore, when clofazimine and dapsone were added together it was observed that the increase of O2- production by macrophages due to clofazimine was not significantly altered by the addition of dapsone . Moreover, it was found that killed Mycobacterium leprae could induce a lesser amount of O2- production in comparison to that of Staphylococcus aureus and the enhancement of O2- release due to clofazimine was stimulus dependent . This increase of O2- release after addition of clofazimine was inhibited by the addition of p-bromophenacyl bromide . Another interesting finding was that the enhancement of O2- production by clofazimine gradually decreased as clofazimine was exposed to light for days . On further investigation it was found that ultraviolet, NMR, infrared and mass spectra of the light unexposed and exposed drug were similar, but the diffusion current of the polarogram of light exposed drug was remarkably more than that observed in light unexposed drug, indicating, thereby, a possible increase in the electron accepting capacity of the light reacted molecule . As far as we know this is the first report describing the effect of light exposed clofazimine on the respiratory burst activity of macrophages.

SAAS Bull Biochem Biotechnol, 1991 Jan, 4, 60 - 7
Effect of cytokines on bovine mammary gland immunity; Nickerson SC; Cytokines are a family of glycoproteins produced by various cell types in response to specific stimuli that regulate the immune response . This paper reviews recent studies on two different cytokines, each with its own effector cell type: interleukin-2 (IL-2), which regulates lymphoid cell responses; and granulocyte colony-stimulating factor (GCSF), which regulates neutrophil responses . In the first study, administration of IL-2 to the bovine mammary gland was found to stimulate the expansion of lymphocyte populations and increase local antibody production . In the second study, systemic administration of GCSF increased peripheral blood as well as milk neutrophil populations, which afforded some protection against Staphylococcus aureus challenge . Results suggest a role for cytokines in the control of mastitis in dairy cattle.

Phytochemistry, 1991, 30(11), 3519 - 23
Isolation and partial amino acid sequence of domains of nitrate reductase from spinach; Fido RJ; Fragments of spinach nitrate reductase (NR) were prepared by limited proteolysis of immunopurified enzyme using both Staphylococcus aureus V8 protease and trypsin . Incubation of NR with V8 protease yielded two enzymically active fragments which could be size separated by FPLC on a Superose 12 column or subjected to further proteolysis while bound to a blue Sepharose affinity column . An NADH-ferricyanide (NADH-FR) active fragment bound to, and was eluted from, a blue Sepharose column by micromolar concentrations of NADH . A fragment with methyl viologen-NR activity was either eluted from the same column using 1 M KNO3 or on further treatment in situ on the blue Sepharose column with trypsin . Incubation of holo-NR with trypsin resulted in the loss of all terminal nitrate reducing activities but no loss in either NADH-FR activity or NADH-cytochrome c reductase activity . Two protease-sensitive regions of NR are shown which connect essentially between the flavin (FAD) and haem domains, and between the haem and molybdenum domains of NR . Amino acid analysis of the FAD- and FAD/haem-containing domains yielded two partial sequences which are compared with sequences deduced from complementary DNA (cDNA) of NR from Arabidopsis, tobacco and spinach . The deduced sequences from Arabidopsis and tobacco are found to be ca 80% and the spinach 100% homologous to the sequence obtained for spinach NR fragments.

J Immunol Methods, 1990 Dec 31, 135(1-2), 33 - 41
A heterogeneous immunoassay performed on a rotating carbon disk electrode with electrocatalytic detection . Mass transfer control of the capture of an enterotoxin; Huet D et al.; An ELISA procedure for the determination of enterotoxin A from Staphylococcus aureus conducted on the surface of a glassy carbon electrode is described . The electrocatalytic detection of the immobilized labelled second antibody is based upon the electrochemical reaction and the enzymatic catalysis occurring on the same surface . The indirect quantification of the bound antigen is, therefore, very sensitive (10(-15) mol cm-2) . This heterogeneous technique was used to study the kinetics of antigen binding to the immunological solid phase, the mass transfer of the antigen being controlled under well-defined hydrodynamic conditions . The experiments were performed with a rotating solid phase disk in such a way that thickness of the diffusion layer was known . We found that the capture of the antigen by the immobilized monoclonal antibody was solely limited by diffusion . A simple theoretical model permitted the amount of bound antigen and the sensitivity of the method to be predicted as a function of the incubation time, the rotational speed of the solid phase and the volume of the sample . Both the theory and the experimental results indicate that the assay may be performed with the sample volume undefined.

Harefuah, 1990 Dec 16, 119(12), 425 - 7
{Intracranial hemorrhage complicating acute disseminated staphylococcal disease in a child}; Goldfarb D et al.; Acute disseminated staphylococcal disease may develop in previously healthy children below the age of 15 years . It progresses rapidly and may cause death in a significant number . The diagnostic criteria are infection in 2 or more anatomical sites and isolation of a coagulase-positive Staphylococcus aureus from the blood or from a site of infection . We present an 11.5-year-old boy with disseminated staphylococcal disease with evidence of cellulitis, osteomyelitis and endocarditis . He developed intracranial hemorrhage as a complication and survived, but with mild residual hemiparesis . Nervous system involvement, such as meningitis and brain abscess, have been described in this particularly severe disease . This is the only known report of intracranial hemorrhage as a complication of the disease.

J Immunol, 1990 Dec 15, 145(12), 3949 - 55
Functional studies examining the subpopulation of human B lymphocytes responding to high molecular weight B cell growth factor; Ambrus JL Jr et al.; Mature human B lymphocytes perform many functions including antibody secretion, Ag presentation, preservation of memory for Ag, and lymphokine secretion . Individual resting B cells receive multiple sequential signals that determine the function(s) that will be performed by those cells . Activation signals such as Ag or Staphylococcus aureus Cowan I (Sac) stimulate overlapping but different subpopulations of B cells . After activation, B cells may be induced to proliferate by a variety of B cell growth factors (BCGF) including IL-2, IL-4, TNF-alpha, low molecular weight BCGF (LMW-BCGF), and high molecular weight BCGF (HMW-BCGF) . Little information exists to explain why so many different BCGFs are involved with human B cell proliferation . The current studies were designed to examine the role HMW-BCGF plays in selecting B cells for particular functions . HMW-BCGF but not LMW-BCGF was found to inhibit Ig secretion when it was included in culture with Sac-activated B cells and B cell differentiation factors (BCDFs) including IL-6 . Sorting resting B lymphocytes into surface IgD+ and IgD- populations and then stimulating each population with anti-mu revealed that the cells most responsive to HMW-BCGF resided in the surface IgD- sorted population . Sorting activated B lymphocytes into BA5 (HMW-BCGFR)+ and BA5- populations revealed that BA5+ B cells stimulated with BCDF (in the absence of HMW-BCGF) produced predominantly IgG, whereas the BA5- population produced both IgG and IgM . Finally, expansion of peripheral B cells from tetanus toxoid-immunized donors with either HMW-BCGF or LMW-BCGF revealed that the HMW-BCGF-expanded population produced predominantly IgG tetanus-specific antibody in the presence of BCDF (in the absence of HMW-BCGF), whereas the LMW-BCGF-expanded population produced IgM much greater than IgG tetanus-specific antibody . Thus, HMW-BCGF may function to expand a subpopulation of B cells for memory B cell functions.

Int J Cancer, 1990 Dec 15, 46(6), 1107 - 11
Establishment and characterization of a new human B-cell line (ONHL-1) from non-Hodgkin's lymphoma: constant expression of bcl-2 gene during mitogen-induced growth inhibition; Matsumura I et al.; A new B-cell line (ONHL-1) was established from non-Hodgkin's lymphoma . ONHL-1 was free from Epstein-Barr virus nuclear antigen and expressed CD20, CD24, and slg (mu, delta, gamma and kappa), thus being equivalent to the mature B-cell stage . Chromosome analysis revealed a markedly abnormal pattern including 14q+ and 6q- . In accordance with the positive expression of surface kappa light chains, one of the kappa genes was found to be rearranged . However, rearrangement of the lambda locus was also detected, contrary to the supposed hierarchy for the rearrangement of the light-chain genes . Further, the rearranged fragments of the JH, C lambda, and bcl-2 genes were of the same size in the EcoRI and HindIII digests on the same filter . This may suggest that the bcl-2 gene is juxtaposed with the JH and C lambda locus . The proliferation of ONHL-I was inhibited by adding Staphylococcus aureus Cowan 1 or 12-O-tetradecanoyl-phorbol-13-acetate . During this growth inhibition, the expression of c-myc decreased, while that of bcl-2 mRNA remained steady . This result suggests that not the bcl-2 gene but other oncogenes, such as c-myc, play a key role in the proliferation of ONHL-1 . This agrees with the hypothesis that the bcl-2 gene is not concerned with aggressive proliferation but with cell survival . This new cell line will therefore be of value in studying the differentiation and tumorigenesis of B cells.

Biochim Biophys Acta, 1990 Dec 14, 1030(2), 251 - 7
Rapid turn-over of plasma membrane sphingomyelin and cholesterol in baby hamster kidney cells after exposure to sphingomyelinase; Slotte JP et al.; Plasma membrane sphingomyelin in baby hamster kidney (BHK-21) cells was hydrolyzed with sphingomyelinase (Staphylococcus aureus) and the effects on membrane cholesterol translocation and the properties of membrane bound adenylate cyclase and Na+/K(+)-ATPase were determined . Exposure of confluent BHK-21 cells to 0.1 U/ml of sphingomyelinase led to the degradation (at 37 degrees C) of about 60% of cell sphingomyelin . No simultaneous hydrolysis of phosphatidylcholine occurred . The hydrolysis of sphingomyelin subsequently led to the translocation (within 40 min) of about 50-60% of cell {3H}cholesterol from a cholesterol oxidase susceptible pool to an oxidase resistant compartment . The translocation of {3H}cholesterol from the cell surface to intracellular membranes was accompanied by a paralleled increase in {3H}cholesterol ester formation . When cells were first exposed to sphingomyelinase (to degrade sphingomyelin) and then incubated without the enzyme in serum-free media, the mass of cell sphingomyelin decreased initially (by 60%), but then began to increase and reached control levels within 3-4 h . The rapid re-synthesis of sphingomyelin was accompanied by an equally rapid normalization of cell {3H}cholesterol distribution . The re-formation of cell sphingomyelin also led to a decreased content of cellular {3H}cholesterol esters, indicating that unesterified {3H}cholesterol was pulled out of the cholesterol ester cycle and transported to the cell surface . Exposure of BHK-21 cells to sphingomyelinase further led to a dramatically decreased activity of ouabain-sensitive Na+/K(+)-ATPase, whereas forskolin-stimulated adenylate cyclase activity was not affected . The activity of Na+/K(+)-ATPase returned to normal in parallel with the normalization of cell sphingomyelin mass and cholesterol distribution . We conclude that sphingomyelin has profound effects on the steady-state distribution of cell cholesterol, and that manipulations of cell sphingomyelin levels directly and reversibly affects the apparent distribution of cholesterol . Changes in the lipid composition of the plasma membrane also appears to selectively affect important metabolic reactions in that compartment.

Eur J Biochem, 1990 Dec 12, 194(2), 491 - 8
Characterization of human factor VIII and interaction with von Willebrand factor . An electron microscopic study; Heijnen HF et al.; Blood coagulation factor VIII is a large glycoprotein that circulates in plasma at relative low concentration (0.1 microgram/ml) . It consists of a heterogeneous mixture of a series heavy-chain peptides (90-200 kDa), each associated with a light chain of 80 kDa . To gain insight into the physical properties of the protein, we have characterized purified human factor VIII by electron microscopy and rotary shadowing . Electron microscopy of rotary shadowed factor VIII molecules showed predominantly a single globular domain structure, with a somewhat asymmetric shape, while two-domain structures were also encountered . The overall dimensions of the globular domains ranged from 4 x 6 nm to 8 x 12 nm . EDTA treatment of factor VIII reduced the overall dimensions (2.5 x 5 nm to 6 x 10 nm) while treatment with thrombin reduced the dimensions to a small extent . In complexes with von Willebrand factor, factor VIII appeared localized at the globular domains of von Willebrand factor multimers . In addition, incubation of factor VIII with Staphylococcus aureus V8 protease fragments SpII and SpIII revealed only binding to the globular domains of SpIII . In this study, the first morphological characterization of human factor VIII is presented, together with its direct localization on von Willebrand factor multimers.

J Biol Chem, 1990 Dec 5, 265(34), 21099 - 107
Phagosomal acidification is mediated by a vacuolar-type H(+)-ATPase in murine macrophages; Lukacs GL et al.; The mechanism underlying phagosomal acidification was studied in thioglycolate-elicited murine macrophages . The pH of the phagosomal compartment (pHp) was measured fluorimetrically in macrophage suspensions following ingestion of fluorescein isothiocyanate-labeled Staphylococcus aureus . At 37 degrees C, pHp decreased rapidly, reaching a steady state value of 5.8-6.1, while the cytoplasmic pH remained near neutrality, pH 7.1 . The phagosome to cytosol pH gradient could be collapsed by addition of nigericin, monensin, or weak bases . The substrate dependence and inhibitor sensitivity profile of phagosomal acidification were investigated in intact and permeabilized cells . Phagosomal acidification was inhibited when ATP was depleted using metabolic inhibitors or permeabilizing the plasma membrane by electroporation . In permeabilized cells, acidification could be initiated by readdition of both Mg2+ and ATP . Neither adenosine 5'-(beta,gamma-imido)triphosphate nor adenosine 5'-(gamma-thio)triphosphate supported phagosomal acidification . Inhibitors of F1F0-type H(+)-ATPase such as oligomycin and azide, and the E1E2-type H(+)-ATPase inhibitor vanadate had no effect on phagosomal acidification . In contrast, the rate of phagosomal acidification was reduced by micromolar concentrations of N-ethylmaleimide and N,N'-dicyclohexylcarbodiimide . In permeabilized cells, nitrate inhibited the acidification with an apparent Ki of 25 mM . Phagosomal acidification was also effectively blocked by the macrolide antibiotic bafilomycin A1, with an apparent Ki of approximately 3 mM in both intact and electroporated cells . In this concentration range, bafilomycin A1 selectively inhibits vacuolar H(+)-ATPases . The substrate requirement and inhibitor susceptibility profile of phagosomal acidification strongly suggest that proton translocation across the phagosomal membrane is mediated by a vacuolar-type H(+)-ATPase.

J Biol Chem, 1990 Dec 5, 265(34), 20739 - 46
Conformation of apolipoprotein B-100 in the low density lipoproteins of tangier disease . Identification of localized conformational response to triglyceride content; Kunitake ST et al.; The low density lipoproteins (LDL) from patients with Tangier disease are enriched in triglycerides, 27% of LDL mass versus 7% for normal LDL . To study whether this unique LDL core lipid composition affects the surface disposition of apolipoprotein (apo) B-100, we analyzed the LDL by protease digestion and in competitive radioimmunoassays . Limited proteolytic digestion of Tangier LDL by Staphylococcus aureus V8 protease generated a prominent fragment of 120 kDa (cleavage site at residue 1076), which was not visible in similarly digested normal LDL . In competitive radioimmunoassay, Tangier LDL bound weakly to the apoB-specific monoclonal antibody MB20, compared with control LDL . We localized the MB20 epitope between residues 1031 and 1084 of apoB-100, probably very near residue 1076 . DNA sequencing of exon 21 of apoB genomic clones (coding for residues 1014-1084) from a Tangier patient revealed no difference from the normal DNA sequence, thus eliminating a protein polymorphism as a basis for the altered protease sensitivity and antibody binding . When the triglyceride contents of Tangier LDL were reduced to 10% of mass by incubation with normal high density lipoproteins, production of the 120-kDa fragment by proteolysis decreased and MB20 binding increased in affinity, implying a change toward normal conformation of apoB-100 . Thus, using two independent techniques, proteolytic digestion and binding of monoclonal antibodies, we have demonstrated an alternative conformation of apoB-100 in the vicinity of residue 1076, which reflects the content of triglycerides in the LDL particle.

Ugeskr Laeger, 1990 Dec 3, 152(49), 3699 - 702
{Mediastinal infection following open heart surgery}; Bjerno T et al.; A retrospective investigation was undertaken of the mediastinal infections in patients submitted to cardiac surgery . A total of 1,763 patients participated in a period of 11 years from 1978 to 1988 . All of the case records were reviewed for the occurrence of infections in the mediastinum and 24 patients (1.4%) were found to have had mediastinitis . Predisposing factors to mediastinal infections are reviewed . In all of the cases, treatment consisted re-thoracotomy with removal of all necrotic and infectious material . This was followed by primary closure over a retrosternal irrigation system which supplied an antibiotic solution for six days . In addition, oral antibiotics were administered on the basis of the findings on culture . The incidence of infection was found to be increased when the left internal mammary artery (LIMA) had been employed and in cases of combined cusp and coronary surgery . In 20 patients (83%), relevant culture findings were found from the mediastinal drain on an average of 2.5 days after the primary operation . These were reencountered in the mediastinum at rethoracotomy which was performed on an average of 15 days after the primary operation . This raises the question of the possibility of earlier intervention . The commonest microorganisms were micrococci and Staphylococcus aureus which together comprized approximately 83% . The antibiotic treatment routinely employed was 1 g meticillin four times daily . Five patients died (21%) and 19 patients were discharged . On follow-up examination, the sternum was stable and the scar healed . When infection in the mediastinum is suspected after cardiac surgery, early diagnosis and active surgical treatment are important . The closed method of irrigation drainage is recommended by the authors.

Nippon Kyobu Geka Gakkai Zasshi, 1990 Dec, 38(12), 2446 - 50
{Management of acute empyema with leakage of bronchial anastomosis after right sleeve upper lobectomy--a case report}; Kondo D et al.; Bronchopleural fistula developing empyema after pulmonary resection is a fatal complication . But we have successfully treated this complication after right sleeve upper lobectomy . The patient was a 61-year-old male, who has a squamous cell carcinoma originating in the right upper lobe of the lung in the clinical stage of IIIa . Right sleeve upper lobectomy with pericardiotomy to reduce the tension of bronchial anastomosis and R2a lymph node dissection . The leakage of bronchial anastomosis was complicated on 5th postoperative day and developed aspiration pneumonia of right middle and lower lobe and empyema . Reoperation was done on 7POD and completion pneumonectomy, omentopexy and open window thoracotomy were performed . Then endotracheal tube had been inserted into the left main bronchus and he controlled under respirator . Six weeks after that fistula was cured . Empyema, which caused by methicillin resistant staphylococcus aureus, was cured by open drainage for 2 weeks and closed drainage and irrigation for 14 weeks . He discharged 16 weeks after reoperation.

Gen Comp Endocrinol, 1990 Dec, 80(3), 363 - 71
The complete amino acid sequence of prolactin from the sea turtle (Chelonia mydas); Yasuda A et al.; The complete amino acid sequence of prolactin (PRL) from a reptile, the sea turtle (Chelonia mydas), was determined for the first time . Sequence analysis was performed on fragments obtained from cleavage of intact and performic acid-oxidized hormone with lysyl endopeptidase, Staphylococcus aureus protease, and o-iodosobenzoic acid employing manual Edman degradation . The sea turtle PRL consists of 198 amino acid residues with three disulfide linkages formed between residues 4-11, 58-173, and 190-198 and possesses heterogeneity indicated by four replacements at positions 55, 145, 148, and 171 . Sequence comparison with other vertebrate PRLs revealed that the degree of sequence identity conforms well to expectations based on phylogeny except for the rodent PRLs; sea turtle PRL has 86% identity with chicken PRL; 81% with horse, pig, and fin whale PRLs; 75-71% with cattle, sheep, and human PRLs; 60-56% with mouse and rat PRLs; and 35-31% with carp, salmon, and tilapia PRLs.

J Appl Bacteriol, 1990 Dec, 69(6), 834 - 44
Adherence of Staphylococcus aureus to cell monolayers; Wyatt JE et al.; Adherence of four strains of Staphylococcus aureus to eukaryotic cell monolayers was assayed with {3H}-thymidine labelled bacterial cells and the results were analysed by non-parametric statistical tests . Adherence to primary (human mesothelial) and semi-continuous (human embryonic lung) cell monolayers was significantly better than to continuous cell lines (HEp2, HeLa and Vero) . HEp2 cell monolayers provided the most reliable assay substrate of the continuous cell lines tested . Variation occurred between bacterial culture batches but the assay measured significant differences between adhesion levels of the strains and distinguished between high level (RN92, 8325-4) and low level (Wood46, ISP458) adhering strains . Adherence to different batches of cell monolayers also varied but relative adherence values for strains were similar and the ranking of strains according to adhesion values was unchanged . Potential adhesion mediators have been monitored for their effect on adhesion of a highly adherent strain (RN92) to HEp2 monolayers . Fibronectin, protein A and anti-protein A did not significantly affect adhesion . Lipoteichoic acid caused a significant inhibition of adhesion . With critical statistical analysis to accommodate inherent variations, this assay provides a useful model to study factors involved in adherence of Staph . aureus to eukaryotic cells.

Int J Food Microbiol, 1990 Dec, 11(3-4), 251 - 7
The use of DNA probes for confirming enterotoxin production by Staphylococcus aureus and micrococci; Ewald S et al.; DNA-DNA colony hybridization was employed to evaluate the results obtained by different immunological methods for detection of staphylococcal enterotoxin . Staphylococcus aureus strains tested for staphylococcal enterotoxin production by immuno-assays and micrococci not previously tested for staphylococcal enterotoxin production were examined for presence of the genes encoding for staphylococcal enterotoxin A, B, C and E by using three corresponding DNA probes . The staphylococcal enterotoxin A probe also detected staphylococcal enterotoxin E gene because of 100% homology . The optimal sensitivity plate method showed the best accordance between the immuno-assay and the hybridization reactions . The enzyme-linked immunosorbent assay detected 12.5 to 17% staphylococcal enterotoxin producers without hybridization reactions . The microslide gel double diffusion test and the reversed passive latex agglutination test showed rather poor accordance with the hybridization reactions . All 17 strains of different micrococci investigated were negative in hybridization with all three DNA probes.

Minerva Med, 1990 Dec, 81(12), 905 - 8
{Multiple retrofascial abscesses caused by staphylococcal sepsis}; Cornacchiari M et al.; A case of multiple retrofacial abscesses (left psoas and left paraspinal muscles) due to septic embolization probably arising from a staphylococcus aureus infection of a phlebopathic ulcer of the leg is reported . The Authors stress the pathogenetic rarity of the clinical picture, the usefulness of computed tomography for a correct diagnosis and for the follow-up of the treatment's efficacy, the favourable clinical course with medical therapy.

J Clin Microbiol, 1990 Dec, 28(12), 2612 - 5
Expression of type 8 capsular polysaccharide and production of toxic shock syndrome toxin 1 are associated among vaginal isolates of Staphylococcus aureus; Lee JC et al.; A colony immunoblot method was developed for serotyping the capsular polysaccharides expressed by Staphylococcus aureus isolates . The method was rapid and specific and was performed with either polyclonal or monoclonal antibodies specific for each of the capsule types . S . aureus isolates were obtained from patients with toxic shock syndrome (TSS) or other staphylococcal infections and from asymptomatic women with vaginal colonization . Among the vaginal isolates of S . aureus, expression of the type 8 capsule was significantly (P less than 0.001) more frequent among strains that produced TSS toxin 1 (TSST-1) than it was among TSST-1-negative strains . In contrast, the frequency of type 8 capsule expression was similar among both TSST-1-positive and -negative strains of S . aureus from patients with nonvaginal TSS . When all vaginal and nonvaginal isolates were compared, TSST-1-negative S . aureus strains were equally distributed among the type 5 and 8 and nontypeable capsule groups, whereas TSST-1-positive strains were predominantly capsule type 8.

Infect Control Hosp Epidemiol, 1990 Dec, 11(12), 639 - 42
Increasing prevalence of methicillin-resistant Staphylococcus aureus in the United States; Boyce JM; In the period 1975 to 1981, methicillin-resistant Staphylococcus aureus (MRSA) emerged as an important nosocomial pathogen in tertiary care centers in the United States . To determine if the prevalence of this organism has continued to increase, a questionnaire was sent to hospital epidemiologists in 360 acute care hospitals . A total of 256 (71%) of the 360 individuals responded . Overall, 97% (246/256) of responding hospitals reported having patients with MRSA in the period 1987 through 1989 . Respondents in 217 hospitals provided estimates of the number of cases seen in 1987, 1988 and 1989 . The percentage of respondents reporting one or more patients with MRSA increased from 88% in 1987 to 96.3% in 1989 (p = .0008) . The percent of respondents reporting large numbers (greater than or equal to 50) of cases per year increased from 18% in 1987 to 32% in 1989 (p = .0006) . Increasing frequency of large outbreaks was observed in community, community-teaching, federal, municipal and university hospitals.

Infect Immun, 1990 Dec, 58(12), 3996 - 4003
Production of tumor necrosis factors alpha and beta by human mononuclear leukocytes stimulated with mitogens, bacteria, and malarial parasites; Ferrante A et al.; Tumor necrosis factors alpha and beta (TNF-alpha and TNF-beta) are multifaceted polypeptide cytokines which may mediate some of the significant changes in cellular homeostasis which accompany the invasion of the mammalian host by viruses, bacteria, and parasites . Although it is well established that bacterial lipopolysaccharide is a potent inducer of TNF-alpha, there is still very little known of the types of agents which can trigger the production of TNFs in mononuclear leukocytes . Using an enzyme-linked immunosorbent assay for measuring TNF-alpha and TNF-beta, we examined the capacity of various T-lymphocyte and beta-lymphocyte mitogens as well as microbial components to stimulate production of these cytokines in culture . The mitogens phytohemagglutinin, concanavalin A, and pokeweed mitogen induced production of both TNF-alpha and TNF-beta, while whole-killed Staphylococcus aureus and Bordetella pertussis, like lipopolysaccharide, were potent inducers of TNF-alpha but failed to stimulate TNF-beta production . TNF-alpha production was detectable within 1 h after stimulation, while TNF-beta production was not detected until after 8 h of culture . The bacterial products tetanus toxoid, purified protein derivative, pertussis filamentous hemagglutinin, and pertussis toxin were all able to induce TNF-alpha and TNF-beta production . Disrupted (frozen-thawed) Plasmodium falciparum-infected erythrocytes were also potent inducers of TNF-alpha and TNF-beta . The results demonstrated that a wide variety of microbial components are inducers of TNF-alpha . Some may not only be more effective than lipopolysaccharide but can also induce TNF-beta production . Furthermore, evidence is presented showing that TNF-beta but not TNF-alpha production correlates with lymphoproliferation.

Am Rev Respir Dis, 1990 Dec, 142(6 Pt 1), 1320 - 4
Risk factors for Staphylococcus aureus nosocomial pneumonia in critically ill patients; Rello J et al.; Staphylococcus aureus nosocomial pneumonia has become an important infection not only because of an apparently increasing incidence but also because of its high mortality rate . A total of 50 episodes of nosocomial pneumonia in critically ill patients in which etiologic diagnosis was well established were prospectively followed in a medical-surgical intensive care unit (ICU) . S . aureus was isolated in a total of 13 episodes . In the univariate analysis the variables significantly associated with S . aureus nosocomial pneumonia were below 25 yr of age, coma, nonuse of corticosteroids, and antecedent trauma . A step-forward logistic regression analysis defined only coma as significantly influencing the risk of developing S . aureus nosocomial pneumonia . We suggest that antimicrobial drugs active against S . aureus must be included in the initial empirical antimicrobial regimen for treating nosocomial pneumonia in patients with coma . The identification of factors influencing the etiology and the possibility of earlier effective antimicrobial treatment may represent a further step in the control of nosocomial pneumonia in critically ill patients by improving its prognosis.

Epidemiol Infect, 1990 Dec, 105(3), 493 - 500
Crystal violet reactions of fresh clinical isolates of Staphylococcus aureus from two British hospitals; Freeman R et al.; When 168 fresh clinical isolates of Staphylococcus aureus were examined for their reactions on a medium containing 1 part in 100,000 crystal violet 50.6% of strains produced a purple appearance, 39.3% produced a white appearance and 10.1% produced a yellow appearance . Purple-reacting isolates were significantly associated with both invasive infections (P less than 0.01) and hospital origin (P less than 0.001) . There were no significant associations between the crystal violet reactions and either animal contact or other properties previously reported to be characteristic of white and yellow-reacting strains (beta haemolysin and bovine coagulase production) . The results of phage typing showed associations between susceptibility to group III phages and purple-reacting strains and between phage group II susceptibility and white and yellow-reacting strains . There was also a highly significant association between white reactions on crystal violet agar and susceptibility to lysis by a combination of all three groups (that is, I + II + III) and white-reacting strains were significantly more susceptible to lysis by phages 94 and/or 96, whether as a restricted pattern or as part of a broader pattern . The purple reaction on crystal violet medium may be a reliable marker of the 'hospital staphylococcus'.

J Infect Dis, 1990 Dec, 162(6), 1400 - 2
Nasal carriage of Staphylococcus aureus: correlation with hormonal status in women; Winkler J et al.; In view of recent observations on hormone-microorganism interactions, a study of Staphylococcus aureus nasal carriage in relation to sex-hormone status was undertaken . Prospectively in 479 women attending a colpocytologic clinic, hormonal status was assessed by determining the karyopyknotic index (KI) on smears stained by the Papanicolaou method . Rates of S . aureus nasal carriage were 29.3% in premenopausal women and 21.9% in postmenopausal women (P not significant) . Carriage rates were significantly higher (P = .026, chi 2 7.32) for women with high KIs (40.7%) than for those with intermediate and low KIs (27.03% and 25.1%, respectively) . S . aureus nasal carriage also correlated independently and significantly with previous antibiotic use and the presence of insulin-treated diabetes mellitus . This preliminary observation confirms an association between levels of sex hormones as reflected by the KI and S . aureus nasal carriage rates.

J Med Microbiol, 1990 Dec, 33(4), 227 - 34
Excision of a conjugative plasmid from the staphylococcal chromosome; Udo EE et al.; Staphylococcus aureus isolate WBG1003 resistant to benzyl penicillin, cadmium, arsenate and streptomycin harbours two plasmids of 38.8 (pWBG621) and 4.4 (pWBG625) kb . In conjugation experiments two types of streptomycin-resistant transconjugants were obtained; one carried a 4.4-kb plasmid and the other, a 34.5-kb and a 4.4-kb plasmid . The 34.5-kb plasmid (pWBG620) has been found to be conjugative and able to mobilise non-conjugative plasmids . It has no detectable resistance phenotype and has not been detected in WBG1003 nor in the recipient used in the conjugation experiments . Restriction endonuclease analysis and DNA-DNA hybridisation have revealed that pWBG620 is unrelated to pWBG621 present in strain WBG1003 . The data presented indicate that pWBG620 is in the chromosome of strain WBG1003 and that it excises during conjugation.

J Bacteriol, 1990 Dec, 172(12), 7260 - 2
DNA gyrase gyrA mutations in ciprofloxacin-resistant strains of Staphylococcus aureus: close similarity with quinolone resistance mutations in Escherichia coli; Sreedharan S et al.; The gyrA genes isolated from three ciprofloxacin-resistant clinical isolates of Staphylococcus aureus carried codon 84 (serine----leucine) and/or codon 85 (serine----proline) mutations that were absent in pretreatment susceptible strains . These substitutions occur in a region of the gyrase A protein wherein directly analogous mutations of serine 83----leucine and alanine 84----proline in Escherichia coli confer quinolone resistance . Thus, DNA gyrase A subunit mutations are implicated in resistance to ciprofloxacin in S . aureus.

J Bacteriol, 1990 Dec, 172(12), 6942 - 9
Nucleotide sequence and characterization of the Staphylococcus aureus norA gene, which confers resistance to quinolones; Yoshida H et al.; The norA gene cloned from chromosomal DNA of quinolone-resistant Staphylococcus aureus TK2566 conferred relatively high resistance to hydrophilic quinolones such as norfloxacin, enoxacin, ofloxacin, and ciprofloxacin, but only low or no resistance at all to hydrophobic ones such as nalidixic acid, oxolinic acid, and sparfloxacin in S . aureus and Escherichia coli . The 2.7-kb DNA fragment containing the norA gene had a long open reading frame coding for 388 amino acid residues with a molecular weight of 42,265, which was consistent with the experimental value of about 49,000 obtained on DNA-directed translation . The deduced NorA polypeptide has 12 hydrophobic membrane-spanning regions and is partly homologous to tetracycline resistance protein and sugar transport proteins . The uptake of a hydrophilic quinolone, enoxacin, by S . aureus harboring a plasmid carrying the norA gene was about 50% that by the parent strain lacking the plasmid, but it increased to almost the same level as that by the latter strain with carbonyl cyanide m-chlorophenyl hydrazone . On the other hand, the uptake of a hydrophobic quinolone, sparfloxacin, was similar in the two strains . These results suggest that the NorA polypeptide may constitute a membrane-associated active efflux pump of hydrophilic quinolones.

J Biochem (Tokyo), 1990 Dec, 108(6), 909 - 13
Regulatory mechanism by the phosphorylation of 20-kDa light chain of porcine aorta smooth muscle myosin; Hasegawa Y et al.; Porcine aorta smooth muscle myosin was subjected to limited proteolysis by Staphylococcus aureus protease (V8-protease) at 30 mM KCl, under which condition the myosin is in the filamentous form . The heavy chain of the myosin molecule was mainly digested at the 68-160 kDa junction, which corresponds to the 50-20-kDa junction in the heavy chain of skeletal muscle myosin subfragment-1 (S-1) . When the filamentous myosin formed a rigor complex in the presence of F-actin, this site was blocked, and the junction between S-1 and subfragment-2 (S-2) was in turn digested specifically . Both phosphorylated and unphosphorylated 20-kDa light chain (LC20) in the aorta myosin remained intact under these conditions . The actin-activated ATPase activity of phosphorylated myosin was not influenced by the cleavage of the S-1-S-2 junction . With unphosphorylated myosin, however, the actin-activated ATPase activity increased with the cleavage of the S-1-S-2 junction and reached the level of ATPase activity of phosphorylated myosin at the stage of complete cleavage . The increase of ATPase activity was found to be proportional to the loss of double-headed myosin . The overall data indicate that LC20 works to suppress the actin-activated ATPase activity, and the suppression is released by the phosphorylation of LC20 . The presence of two heads in myosin is required to reveal such regulation by LC20.

Immunology, 1990 Dec, 71(4), 473 - 9
Regulation of CD23 expression, soluble CD23 release and immunoglobulin synthesis of peripheral blood lymphocytes by glucocorticoids; Fischer A et al.; Evidence was obtained that glucocorticoids are capable of modulating the CD23 expression and soluble(s) CD23 release of peripheral blood lymphocytes (PBL) . We demonstrate that interleukin-2 (IL-2)- and IL-4-induced CD23 expression are susceptible to glucocorticoids to a different degree . Prednisolone suppressed the spontaneous and IL-2-induced CD23 expression on PBL of healthy donors . The IL-4-induced CD23 expression was influenced much less by prednisolone, but the expression kinetics was altered . The modulation of the expression kinetics appears to be due to a priming effect of prednisolone . Differences were also apparent when the susceptibility of PBL from healthy and atopic donors towards the effect of prednisolone on the IL-4-induced CD23 expression was studied . Preactivation of PBL with Staphylococcus aureus strain Cowan I abolished the differences . Prednisolone also suppressed the sCD23 release from unstimulated and IL-2- or IL-4-stimulated PBL and enhanced the immunoglobulin (E,G,A,M) synthesis of PBL . This enhancement appears to be due to a priming effect, since pre-stimulation of PBL with prednisolone was sufficient to enhance the immunoglobulin synthesis . The IL-4-induced IgE synthesis of PBL with or without spontaneous in vitro IgE synthesis was synergistically enhanced by glucocorticoids.

Zh Mikrobiol Epidemiol Immunobiol, 1990 Dec, (12), 10 - 2
{The quantitative determination of the DNAse activity in Staphylococcus aureus isolated from monkeys}; Kakubava VV et al.; A new, cheaper and more sensitive method for the quantitative determination of DNAase produced by S . aureus is described . The method permits the determination of DNAase activity in a wider range of titers . The method is based on the detection of the depolymerizing action of staphylococcal nuclease on DNA dyed with ethidium bromide . In this work 22 S . aureus strains isolated from monkeys and 12 strains isolated from humans have been used . The amount of produced by these strains has been determined . The DNAase results of this determination have shown that among S . aureus strains isolated from monkeys and humans the occurrence of strains with both high and low DNAase activity can be observed.

Antimicrob Agents Chemother, 1990 Dec, 34(12), 2312 - 7
Treatment of experimental foreign body infection caused by methicillin-resistant Staphylococcus aureus; Lucet JC et al.; A novel model of experimental foreign body infection was developed in rats: four perforated Teflon tissue cages per animal were implanted subcutaneously and 3 to 4 weeks later were infected with 0.5 x 10(5) to 2 x 10(5) CFU of methicillin-resistant Staphylococcus aureus . After 2 weeks, the number of CFU in the cage fluid was determined {day 1 mean, (7.25 +/- 0.79) log10 CFU/ml}, and treatment with vancomycin (50 mg/kg twice a day {BID}), fleroxacin (50 mg/kg BID), or fifampin (25 mg/kg BID), alone and in combination, was initiated for a duration of 6 days . Concentrations of antibiotics in cage fluids were in the range of those encountered in clinical conditions . Eighteen hours after the last injection (day 7), the number of CFU in the cage fluid was determined and the difference between day 1 and day 7 values was calculated . Rifampin, alone and in combination with fleroxacin or vancomycin, was the most effective regimen in reducing the bacterial counts in the tissue cage fluids {(1.87 +/- 1.44, 2.18 +/- 1.02, and 2.55 +/- 1.09 log10) CFU/ml, P less than 0.001, respectively} . After treatment, cage fluids and cages were analyzed for resistant bacteria . Resistance to rifampin occurred in 15 of 19 cages in animals treated with rifampin alone and in 4 of 25 in animals treated with rifampin plus vancomycin . We detected no development of resistance to rifampin in animals treated with rifampin plus fleroxacin or to fleroxacin in animals treated with this antimicrobial agent . In conclusion, regimens including rifampin alone or in combination with vancomycin or fleroxacin were an effective treatment of foreign body infection due to methicillin-resistant S . aureus in reducing bacteria counts, but rifampin monotherapy was compromised by significant emergence of resistance . The combined therapy of fleroxacin with rifampin prevent development of resistance to rifampin.

Zhonghua Jie He He Hu Xi Za Zhi, 1990 Dec, 13(6), 336 - 8, 379
{Pathogenic and clinical changes of pneumonia in the recent 10 years}; Jiang XF; The article reviewed 279 cases of pneumonia which came from two different periods . Among these 116 got from 1960s and 174, from 1980s . We analyzed the general condition of patients, etiology, mortality, prognosis and management and reached to the conclusion that (1) gram-negative bacilli become major pathogen in replace of Gram-positive staphylococcus aureus in 1980s, but staphylococcus aureus pneumonia is still a serious disease (2) the prognosis of the patient is related to underlying disease, nourishment and complication (3) at present, Cephalosporin V is most effective in Gram-positive cocci pneumonia and Gentamycin or Cephalosporin V, in Gram-negative bacilli pneumonia . (4) the mortality rate of pneumonia in 1980s is lower than that in 1960s.

Vet Immunol Immunopathol, 1990 Dec, 26(4), 319 - 32
Chicken macrophage activation by interferon: do birds lack the molecular homologue of mammalian interferon-gamma?
Dijkmans R, Creemers J, Billiau A.
In mammalian species, interferon-gamma (IFN-gamma) is a lymphokine with a wide range of biological effects, of which the antiviral and macrophage-activating capacities are those best characterized . In birds, no equivalent with a similar range of actions has as yet been isolated . Chicken splenocytes were stimulated by mitogens in conditions that were similar to those used for the induction of mammalian IFN-gamma . Culture fluids were assayed for antiviral and macrophage-activating capacities . As much as 1000 units/ml of an interferon-like antiviral activity was found in the culture fluid of Staphylococcus aureus lysate-induced spleen cells . Seroneutralization assays with a polyclonal antiserum against purified interferon and physicochemical studies revealed that the antiviral activity is identical to or closely related to type I interferon (interferon-alpha/beta) . The presence of macrophage activating factors (MAF) in the splenocyte medium was demonstrated by measuring increased production of H2O2 by chicken peritoneal macrophage cultures and a chicken macrophage cell line (HD11) . The heat stability of this MAF activity was similar to that of the antiviral factor, and was completely neutralized by the anti-IFN-alpha/beta antiserum . These results show that when the classical procedure used for the production of mammalian IFN-gamma is applied to chicken splenocytes, it does not yield an equivalent for IFN-gamma/MAF . This suggests that the classification of interferons into types (alpha, beta and gamma), while generally applicable in mammals, may not be applicable in birds.

Malays J Pathol, 1990 Dec, 12(2), 107 - 9
Phage typing of methicillin-resistant Staphylococcus aureus; Abu Hanifah Y; 448 isolates of methicillin-resistant Staphylococcus aureus (MRSA) from clinical specimens of patients from the University Hospital, Kuala Lumpur, were phage-typed . These included 35 strains causing two separate outbreaks of infection, one in surgical Ward 6B and another in the Special Care Nursery (SCN) . Antibiograms of these outbreak strains in Ward 6B and SCN were entirely different . Phage-typing revealed that 72% of the MRSA isolates were typable . They were typed entirely by Group III phages, the majority (76%) of which were phage type 85 . There was only one isolate in SCN which was typed by Group I (phage 80) and Group III phages . None were typed by phages 94, 95, 96 and Group II phages . 14.6% of the typable isolates gave the long pattern reaction of the phage 6/47/54/75/77/83A/84/85 complex . The majority of the outbreak strains in Ward 6B were of phage type 85, whereas those in the SCN were all of the 6/47/54/75/77/83A/84 phage pattern with the exception of one isolate which was also typed by phage 80, a Group I phage.

J Dairy Sci, 1990 Dec, 73(12), 3457 - 62
Protective effect of Staphylococcus chromogenes infection against Staphylococcus aureus infection in the lactating bovine mammary gland; Matthews KR et al.; The susceptibility of uninfected or Staphylococcus chromogenes-infected quarters to challenge with Staphylococcus aureus was measured . Seventeen S . chromogenes-infected quarters were challenged by infusion of S . aureus into the teat sinus; 47% (8 of 17) became infected and all 18 uninfected quarters challenged similarly with S . aureus became infected . No differences in daily milk yield were seen between uninfected quarters and S . chromogenes-infected quarters prior to S . aureus infusion . Postinfusion, milk yield for S . aureus-infected, S . chromogenes-infected, and S . chromogenes- and S . aureus-infected quarters differed . Somatic cell counts were elevated in S . chromogenes-infected quarters compared with uninfected quarters prior to S . aureus infusion . Somatic cell counts were not different between S . aureus- and S . chromogenes- and S . aureus-infected quarters postinfusion, but were different for S . chromogenes-infected quarters . Chloride concentrations in S . chromogenes- and S . aureus-infected quarters were different from either S . aureus-infected or S . chromogenes-infected quarters . Staphylococcus aureus colony forming units in quarters with preexisting S . chromogenes infections were lower than S . aureus colony-forming units in previously uninfected quarters . Possible protective mechanisms induced by S . chromogenes against superinfection by S . aureus are discussed.

J Dairy Sci, 1990 Dec, 73(12), 3449 - 56
Determination of milk and mammary tissue concentrations of ceftiofur after intramammary and intramuscular therapy; Owens WE et al.; Twenty-five Staphylococcus aureus strains isolated from bovine mastitis were tested for their susceptibility to ceftiofur . Zone diameter for 30 micrograms disks averaged 39 mm, and minimum inhibitory concentrations ranged from .5 to 1 microgram/ml . Tissue and milk concentrations were determined from biopsy and quarter milk samples collected from eight cows treated with either intramammary infusion of 100 or 200 mg of ceftiofur, one or two intramuscular injections of 500 mg of ceftiofur, or combination therapy of intramammary infusion coupled with intramuscular injection . Three additional cows received two intramammary infusions of 200 mg of cephapirin at 24-h intervals . Intramuscular injections of ceftiofur resulted in tissue and milk concentrations below detectable limits . Staphylococcus aureus was not eliminated from infected mammary glands by infusion of 100 mg of ceftiofur or by injection of 500 mg of ceftiofur by 48 h after treatment . Combination therapy of 100 mg of ceftiofur infused and 500 mg injected reduced S . aureus numbers in milk and tissue markedly, as did infusion of 200 mg of ceftiofur . Cows receiving intramammary infusion of 200 mg of ceftiofur (two doses at 24-h intervals) had highest concentrations in milk (450 micrograms/ml at 4 and 6 h) and in tissue (.08 microgram/mg at 30 h) . These concentrations are similar to those obtained with two 200-mg doses of cephapirin at 24-h intervals . Histologic analysis of mammary parenchymal tissues showed that combination therapy resulted in higher percentages of alveolar luminal area and lower percentages of interalveolar stroma compared with infusion or injection alone . Histology of quarters receiving combination therapy was not different from that of quarters receiving cephapirin infusion alone.

Zh Mikrobiol Epidemiol Immunobiol, 1990 Dec, (12), 66 - 9
{The comparative characteristics of the immunomodulating properties of protein A from Staphylococcus aureus of different origins}; Kondakov KE et al.; The immunomodulating properties of highly purified staphylococcal protein A and its analog obtained by gene engineering techniques have been compared with those of commercial preparations . The comparison has shown that the differences observed in this investigation may be explained by the presence of admixtures of staphylococcal nature in commercial preparations . The preparations of highly purified staphylococcal and recombinant protein A stimulate humoral immune response and the processes of phagocytosis and do not show mitogenic activity with respect to T cells . The conclusion on the identity of the immunomodulating activity of the preparations of natural and recombinant protein A has been made.

Arq Bras Cardiol, 1990 Dec, 55(6), 385 - 8
{Staphylococcus aureus endocarditis in a puerperal woman with mitral and tricuspid valve prolapse}; Pereira Mde B et al.; A 17-year-old woman with mitral and tricuspid valve prolapse and myxomatous degeneration presented puerperal infection by Staphylococcus aureus with clinical picture of sepsis and multiple septic embolism (right eye, left thumb, spleen, and left calf) . She underwent total hysterectomy on the 10th day postdelivery and right eye enucleation on the 16th . Temporary total AV block occurred on the 14th day with temporary external pacing during the next couple of days . Acute endocarditis with acute mitral regurgitation was diagnosed on the 13th day, demanding immediate valve replacement . On the 46th day she developed moderate tricuspid valve regurgitation due to another episode of endocarditis . Final clinical discharge took place on the 62nd day after antibiotic therapy completion.

Zentralbl Bakteriol, 1990 Dec, 274(3), 333 - 41
Nasal carriage of Staphylococcus aureus and its influence on hospital infections caused by methicillin-resistant strains; Heczko PB et al.; A short review of the literature on the actual epidemiological situation related to nosocomial infections caused by methicillin-resistant Staphylococcus aureus and the importance of the nasal carriage of this pathogen is given together with selected data derived from our own studies on these problems.

Mol Microbiol, 1990 Dec, 4(12), 2051 - 62
Efflux-mediated antiseptic resistance gene qacA from Staphylococcus aureus: common ancestry with tetracycline- and sugar-transport proteins; Rouch DA et al.; Resistance to intercalating dyes (ethidium, acriflavine) and other organic cations, such as quaternary ammonium-type antiseptic compounds, mediated by the Staphylococcus aureus plasmid pSK1 is specified by an energy-dependent export mechanism encoded by the qacA gene . From nucleotide sequence analysis, qacA is predicted to encode a protein of Mr 55017 containing 514 amino acids . The gene is likely to initiate with a CUG codon, and a 36 bp palindrome immediately preceding qacA, along with an upstream reading frame with homology to the TetR repressors, may be components of a regulatory circuit . The putative polypeptide specified by qacA has properties typical of a cytoplasmic membrane protein, and is indicated to be a member of a transport protein family that includes proteins responsible for export-mediated resistance to tetracycline and methylenomycin, and uptake of sugars and quinate . The analysis suggests that N- and C-terminal regions of these proteins are involved in energy coupling (proton translocation) and substrate transport, respectively . The last common ancestor of the qacA and related tet (tetracycline resistance) lineages is inferred to have been repressor controlled, as occurs for modern tet determinants from Gram-negative, but not those from Gram-positive, bacteria.

J Clin Pharm Ther, 1990 Dec, 15(6), 425 - 33
Comparison of cultured human mononuclear cells, Limulus amebocyte lysate and rabbits in the detection of pyrogens; Hansen EW et al.; Isolated human mononuclear cells exposed to either lipopolysaccharide or Staphylococcus aureus secreted interleukin-1 like material . The secretion was concentration dependent . The minimal detectable level in the test solution of lipopolysaccharide and Staphylococcus aureus was 200 pg/ml and 10(5) cells/ml respectively . The sensitivity and specificity of the Limulus Amebocyte Lysate test, the rabbit pyrogen test and the monocyte test are compared . The monocyte test is proposed as an alternative in-vitro test to the rabbit pyrogen test.

J Inorg Biochem, 1990 Dec, 40(4), 349 - 56
Syntheses, characterization, and microbial activity of some transition metal complexes involving potentially active O and N donor heterocyclic ligands; Nagar R; The formation of binary as well as ternary metal complexes of type MLL' (where M(II) = Cu(II), Ni(II), Co(II), and Zn(II); L = 8-hydroxyquinoline, and L' = 2-furoic acid) has been studied . The complexes were synthesized and characterized by elemental analyses, molecular weight determination, the IR and electronic spectra, conductivity, and magnetic measurements . The presence of coordinated water molecules was demonstrated by thermogravimetric analysis . The microbial activity of these ligands and their metal complexes was determined on gram positive (Staphylococcus aureus) and gram negative (Escherichia coli) bacteria, the antifungal activity on some common fungi, viz . Aspergillus niger, Aspergillus nidulense, and Penicillium citrinum.

J Protein Chem, 1990 Dec, 9(6), 695 - 703
Mass spectrometric analysis of rabbit and bovine trypsin-solubilized cytochrome b5; Gibson BW et al.; The sequence and blocking group of the amino-terminal 15 amino acids of rabbit trypsin-solubilized cytochrome b5 were determined by liquid secondary ion mass spectrometry (LSIMS) and tandem mass spectrometry (MS/MS) . The molecular weights of peptides generated from a Staphylococcus aureus V8 protease digest of this protein were determined by LSIMS analysis and the two peptides containing the blocked amino-terminus were sequenced by tandem mass spectrometry to yield the sequence; N-acetyl-Ala-Ala-Glu-Ser-Asp-Lys-Asp-Val-Lys-Tyr-Tyr-Thr-Leu-Glu-Glu . Comparison of this sequence with a recently reported cDNA sequence (Dariush et al., 1988) indicates that Gln at position 3 is selectively deamidated, although no other discrepancies were found . Intact rabbit and bovine trypsin-solubilized cytochrome b5 were also analyzed by LSIMS on a high-field mass spectrometer equipped with a diode array detector . Mass measurement of the unresolved protonated molecular ion peak tops gave average molecular weights of 9462.2 +/- 2 and 9502.3 +/- 2 for bovine and rabbit trypsin-solubilized cytochrome b5, respectively . In both cases, these molecular weights correspond to a cytochrome b5 fragment consisting of amino acids Asp(7)-Arg(88) . The average molecular weight for the rabbit amino-terminal-blocked form of trypsin-solubilized cytochrome b5 was found to be 10,144.5 +/- 2, which was consistent with the molecular weight predicted for the extended N-acetylated form (residues 1-88) of Mr 10,146.1.

Br J Haematol, 1990 Dec, 76(4), 513 - 20
CD4 and CD8 subpopulation changes during high dose intravenous immunoglobulin treatment; Macey MG et al.; High doses of immunoglobulin, when given intravenously (IVIgG), influence lymphocyte subset numbers and function . T-lymphocytes may be subdivided into two functionally different groups, helper/inducer (CD4+) and suppressor/cytotoxic (CD8+) . Considerable functional as well as phenotypic heterogeneity exists within the two major subsets . CD4+ cells have been further subdivided into helper/inducer and suppressor/inducer sets by the differential binding of two monoclonal antibodies 4B4 (CDw29) and 2H4 (CD45R) . Similarly, the CD8+ subset may be subdivided into suppressor and cytotoxic populations by the differential binding of monoclonal antibodies which identify the C3bi receptor (CD11) . During IVIgG treatment of patients with autoimmune thrombocytopenic purpura (ATP) the change in CD4/CD8, due to an absolute increase in CD8+ cells, has been shown to correlate with the response to treatment as determined by platelet increase . However, the total CD4+ and CD8- numbers may not reflect changes in their constituent subpopulations . To examine this possibility the CD4 and CD8 subpopulations were analysed in 15 ATP patients, during IVIgG treatment, using a double fluorescence technique . In 10 of these patients the in vitro response to pokeweed mitogen (PWM) and Staphylococcus aureus Cowan I (STA Cowan I) was determined . There was no correlation between the change in CD8+ subpopulations and response to treatment but there was a correlation between the CD4+ change and platelet increment . In addition there was a correlation between the 4B4/2H4 change and the in vitro response to PWM but no correlation with the response to STA Cowan I . These findings suggest that during IVIgG treatment the CD4+ 4B4+ helper/inducer population is influenced resulting in reduced T-dependent B-cell activation.

Antimicrob Agents Chemother, 1990 Dec, 34(12), 2348 - 53
Comparative efficacy of daptomycin, vancomycin, and cloxacillin for the treatment of Staphylococcus aureus endocarditis in rats and role of test conditions in this determination; Cantoni L et al.; The in vivo efficacy of daptomycin, a new cell wall-active anti-gram-positive-bacterial agent, was compared to those of cloxacillin and vancomycin in a rat model of Staphylococcus aureus endocarditis . Both methicillin-susceptible S . aureus (MSSA) and methicillin-resistant S . aureus (MRSA) strains were used . When therapy was initiated early (8 h) after infection, at the time when valvular bacterial counts were relatively low (approximately 10(6) CFU/g of vegetation), 3 days of therapy was found to be effective against the MSSA strains whatever the antibiotic regimen . In contrast, when the onset of therapy was delayed up to 15 h after infection, so that higher bacterial counts could develop on the valves (approximately 10(9) CFU/g of vegetation), a longer period of treatment (6 days) was required to cure infection . Under these conditions after 3 days of therapy, daptomycin was more effective than cloxacillin and vancomycin against the MSSA strains . Similarly, daptomycin showed a greater activity than vancomycin against the MRSA strain after 3 days of treatment, but after 6 days both antibiotics were equally effective . Decreasing doses of daptomycin showed decreasing activity: 10 mg/kg of body weight every 12 h (q12h) was better than 5 mg/kg q12h, whereas 5 mg/kg q24h (providing drug levels in blood detectable only during the first 12 h) failed to cure infection . In vitro, daptomycin was highly bactericidal at high concentrations (25 and 60 micrograms/ml, corresponding to peak levels in serum after doses of 5 and 10 mg/kg, respectively) and bacteriostatic at lower concentrations (0.5 to 2.5 micrograms/ml, corresponding to trough levels in serum) . In conclusion, against low-bacterial-count S . aureus endocarditis, daptomycin showed an efficacy similar to those of vancomycin and cloxacillin . Against high-bacterial-count S . aureus endocarditis, daptomycin showed a higher bactericidal activity than cloxacillin (against the MSSA strains) and vancomycin (against both the MSSA and MRSA strains).

Eur J Clin Invest, 1990 Dec, 20(6), 620 - 6
Cystic fibrosis patients' B-lymphocyte response is resistant to the in vitro enhancing effect of corticosteroids; Emilie D et al.; Cystic fibrosis is associated with an cAMP-regulated channel defect, which has been evidenced in many cell types including B lymphocytes . To document a B-cell dysfunction potentially related to this defect, we studied the in vitro IgG production by lymphocytes from 11 cystic fibrosis patients . B lymphocytes were co-cultured with autologous monocytes and stimulated with Staphylococcus aureus Cowan or with Nocardia-delipidated cell mitogen in the presence of low concentrations of IL2 . Cystic fibrosis patients' cells produced amounts of IgG comparable with that of normal and control patients' cells . However, dexamethasone (10(-7) mol l-1) had no effect on the response of cystic fibrosis patients' cells, whereas it enhanced that of the latter two groups . This resistance of cystic fibrosis cells was true with concentrations of dexamethasone up to 10(-6) mol l-1, whereas this agent induced a dose-related enhancement from 10(-8) to 10(-6) mol l-1 in cultures of normal cells . Co-culture experiments showed that cystic fibrosis B lymphocytes themselves are resistant to the effect of dexamethasone . In contrast dexamethasone normally suppressed the anti-CD3 antibody-induced response of cystic fibrosis T cells in the presence of IL2 and the IL1 alpha- or beta-induced collagenase production of cystic fibrosis fibroblast cell lines . Thus cystic fibrosis B lymphocytes exhibit a selective defect which may interfere with the normal interactions between the hormonal and immune systems and may participate in the sensitivity of cystic fibrosis patients to bacterial bronchopulmonary infections.

Gen Physiol Biophys, 1990 Dec, 9(6), 569 - 75
Memory is a property of an ion channels pool: ion channels formed by Staphylococcus aureus alpha-toxin; Krasilnikov OV et al.; The short-time depolarization effects on the integral conductance induced by S . aureus alpha-toxin (ST) in planar lipid bilayer membranes has been studied . Ion channels formed by ST were found to have several potential-induced nonconductance (closed) states . The transitions of ion channels between the states are only through one conductance state . The transition of ST-channels from closed to open state is induced by membrane depolarization . The amplitude current after a series of voltage pulses is a function of pulse number, and is effectively independent of the time interval between the neighbouring pulses . Therefore, a membrane which contains a pool of ion channels "remembers" its previous existence . A simple model can be used to explain this phenomenon.

Scand J Immunol, 1990 Dec, 32(6), 631 - 40
The response of human B cells to interleukin 4 is determined by their stage of activation and differentiation; Maher DW et al.; The effect of purified recombinant human interleukin 4 (IL-4) on proliferation and IgM secretion of normal and malignant human B cells was studied . IL-4 was found to co-stimulate the proliferation of splenic B cells in the presence of anti-Ig coupled to polyacrylamide beads (anti-Ig beads) for a period of 4 days . In contrast, IL-4 had little co-stimulatory effect on the proliferative response of splenic B cells to the more potent mitogen Staphylococcus aureus Cowan strain 1 (SAC) . Moreover, IL-4 inhibited interleukin 2 (IL-2)-induced proliferation of cells co-stimulated with SAC . Mitogen-induced pre-activation of B cells in the presence of IL-4 resulted in a reduction in subsequent IL-2-induced IgM secretion without significantly affecting proliferation . Human B-cell tumours were also cultured over a 2-3 day period in the presence of anti-Ig beads plus IL-2, or IL-4 or both IL-2 and IL-4 . IL-4 inhibited IL-2-induced proliferation in all cases of B-cell chronic lymphocytic leukaemia (B-CLL) and the majority of cases of low-grade lymphoma (LGL) and hairy cell leukaemia (HCL) . These findings suggest that IL-4 has stimulatory actions on resting B cells, most evident in the presence of submaximal co-mitogenic signals, and inhibitory actions on activated B cells, especially antagonism of the effects of IL-2.

Eur J Immunol, 1990 Dec, 20(12), 2679 - 84
CD27 expression by a distinct subpopulation of human B lymphocytes; Maurer D et al.; CD27 is present on the surface of a major subset of peripheral blood T lymphocytes . In this report we show that CD27 is also expressed on a subpopulation of the normal human B cell lineage which is absent from cord blood but present in tonsils and in the peripheral blood of adult individuals . CD27+ B lymphocytes are characterized by the following criteria: (a) in terms of physical properties, the CD27+ B cells form a population with an increased cell size combined with a decreased cell density; (b) the CD27 expression of tonsillar B lymphocytes is postively correlated with mIgA but negatively correlated with membrane IgM/membrane IgD positivity; (c) CD27 on B cells can be induced selectively by the combination of Staphylococcus aureus plus interleukin 2, but not by either treatment alone, and (d) CD27+ B lymphocytes express high levels of the adhesion structures LFA-1 (CD11a), ICAM-1 (CD54), LFA-3 (CD58) and of the lymphocyte homing receptor CD44 . These latter findings suggest that CD27+ B cells are predispose to form cell-cell interactions . Accordingly, within 3 h of cell culture CD27+, but not CD27-, B lymphocytes were found to form LFA-1-mediated homotypic B cell clusters.

J Invest Dermatol, 1990 Dec, 95(6), 647 - 52
Monoclonal antibodies to two different epitopes in a 30-kD CNBr peptide of the K1 and K2 keratins; Colbert MC et al.; Two anti-keratin monoclonal antibodies, Kab-2 and Kab-3, with specificities for different epitopes of type II (basic) human epidermal keratins, were produced . These antibodies had different immunofluorescent staining patterns on human fetal epidermis . Western blots and solid phase RIA showed both antibodies bound to 65-67-kD basic keratins (K1 and K2) extracted from foreskin epidermis . Competitive binding studies with the two Kab antibodies and other anti-keratin monoclonal antibodies showed that Kab-2 and Kab-3 recognized related epitopes, distinct from the epitopes recognized by other anti-keratin antibodies AE-1, 2, and 3 . Kab-2 and Kab-3 epitopes were distinguished by differences in their reactivity with peptides generated by Staphylococcus aureus V8 protease digestion of the K1 keratin; the antibodies recognized both common and unique peptides . Western blots of cyanogen bromide digests of the K1 keratin showed that both Kab antibodies reacted with a 30-kD fragment of the molecule presumed to be the N-terminal CNBr peptide . We interpret these data to indicate that in tissues, portions of the N-terminal region of the K1 keratin are differentially available for reaction with these monoclonal antibodies and that morphologic differences in staining with monoclonal antibodies to the same molecule can reflect epitope specificity or epitope availability related to supramolecular organization.

Biochem Biophys Res Commun, 1990 Nov 30, 173(1), 92 - 8
A novel epidermal cell differentiation inhibitor (EDIN): purification and characterization from Staphylococcus aureus; Sugai M et al.; A factor inhibiting the calcium-induced terminal differentiation of cultured mouse keratinocytes was purified to homogeneity from the extracellular products of S . aureus E-1 and designated 'epidermal cell differentiation inhibitor' (EDIN) . EDIN activity was sensitive to trypsin and heat-labile, suggesting that EDIN is a protein . EDIN gave a single band with a molecular weight of 27,000 on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and was found to be a single chain polypeptide, having an isoelectric point higher than 9 . The N-terminal amino acid sequence of EDIN was determined as A-D-V-K-N-F-T-D-L . EDIN inhibited the differentiation of not only mouse but also human keratinocytes in culture.

Eur J Biochem, 1990 Nov 26, 194(1), 19 - 23
Amino acid sequence of a 12-kDa inhibitor of protein kinase C; Mozier NM et al.; The complete primary structure of a bovine-brain-derived inhibitor of protein kinase C has been established . Fragments of the purified protein were obtained by cleavage with cyanogen bromide, Staphylococcus aureus V8 protease, trypsin and chymotrypsin . Subsequent analysis of the resulting fragments by fast-atom-bombardment mass spectrometry and Edman degradation revealed a calculated molecular mass of 11,779 Da with the following 107-amino-acid sequence: {sequence: see text} This inhibitor does not share significant primary structural identity with any other known protein.

Proc R Soc Lond B Biol Sci, 1990 Nov 22, 242(1304), 87 - 90
Sequence similarities between the gene specifying 1-phosphofructokinase (fruK), genes specifying other kinases in Escherichia coli K12, and lacC of Staphylococcus aureus; Orchard LM et al.; The sequence was determined of the 936 nucleotides that compose the fruK gene of Escherichia coli K12, together with the final 310 bases of fruF and the initial 224 bases of fruA, which flank fruK . These genes specify proteins that effect the uptake of fructose and its PEP-dependent conversion to fructose 1-phosphate (fruA and fruF), and the ATP-dependent phosphorylation of that product to fructose 1,6-bisphosphate (fruK); together, these genes form the fruFKA operon . The deduced amino acid sequence of the fruK product exhibits little similarity to the major 6-phosphofructokinase of E . coli (pfkA) and the 6-phosphofructokinases present in a number of pro- and eukaryotic organisms, but there is 27%, 25% and 22% identify of sequence respectively with the minor 6-phosphofructokinase (pfkB) of E . coli, the lacC gene product of Staphylococcus aureus and the ribokinase of E . coli.

Med J Aust, 1990 Nov 19, 153(10), 585 - 7
Leg ulcers in rheumatoid arthritis; Pun YL et al.; A retrospective study over an eight-year period of 33 episodes of leg ulceration in 26 patients with rheumatoid arthritis requiring inpatient management is reported . the aetiology of the ulcers was found to be multifactorial . The most common factors were venous insufficiency (45.5%), trauma or pressure (45.5%) and arterial insufficiency (36.4%) . Vasculitis (18.2%) and Felty's syndrome (12.1%) were less frequent causes, and pyoderma gangrenosum was rare . Most patients had seropositive erosive disease with high rheumatoid factor titres and significant functional impairment; over half were on maintenance corticosteroids . Colonisation of the ulcers by organisms, predominantly Staphylococcus aureus, was common (69.7%) . Skin grafting was required in 63.3%, but the rate of complete take was only 42.9% despite multiple attempts . Hospitalisation was prolonged (mean 47.9 days) and the recurrence rate requiring further hospitalisation was 26.9% . The diagnosis of vasculitis and the limited role of biopsy in establishing its presence are discussed.

Biochem J, 1990 Nov 15, 272(1), 151 - 8
Localization of the forskolin photolabelling site within the monosaccharide transporter of human erythrocytes; Wadzinski BE et al.; Chemical and proteolytic digestion of intact erythrocyte glucose transporter as well as purified transporter protein has been used to localize the derivatization site for the photoaffinity agent 3-{125I}iodo-4-azido-phenethylamino-7-O-succinyldeacetylforskol in {( 125I}IAPS-forskolin) . Comparison of the partial amino acid sequence of the labelled 18 kDa tryptic fragment with the known amino acid sequence for the HepG2 glucose transporter confirmed that the binding site for IAPS-forskolin is between the amino acid residues Glu254 and Tyr456 . Digestion of intact glucose transporter with Pronase suggests that this site is within the membrane bilayer . Digestion of labelled transporter with CNBr generated a major radiolabelled fragment of Mr approximately 5800 putatively identified as residues 365-420 . Isoelectric focusing of Staphylococcus aureus V8 proteinase-treated purified labelled tryptic fragment identified two peptides which likely correspond to amino acid residues 360-380 and 381-393 . The common region for these radiolabelled peptides is the tenth putative transmembrane helix of the erythrocyte glucose transporter, comprising amino acid residues 369-389 . Additional support for this conclusion comes from studies in which {125I}APS-forskolin was photoincorporated into the L-arabinose/H(+)-transport protein of Escherichia coli . Labelling of this transport protein was protected by both cytochalasin B and D-glucose . The region of the erythrocyte glucose transporter thought to be derivatized with IAPS-forskolin contains a tryptophan residue (Trp388) that is conserved in the sequence of the E . coli arabinose-transport protein.

Blood, 1990 Nov 15, 76(10), 2091 - 7
Response patterns of hairy cell leukemia to B-cell mitogens and growth factors; Barut BA et al.; The effect of mitogens and/or recombinant B-cell growth factors (M/GFs) on the in vitro growth of hairy cells was examined . Tumor cells were isolated from the spleens of four patients with hairy cell leukemia (HCL) by Ficoll-Hypaque sedimentation and E-rosetting . Enrichment for tumor cells was confirmed with intracytoplasmic immunoglobulin (Ig) staining, tartrate resistant acid phosphatase (TRAP) staining, and staining using monoclonal antibodies (MoAbs) directed at B, T, myeloid, and monocytoid antigens (Ags) in indirect immunofluorescence assays . Tumor cells were B1(CD20)+ B2(CD21)- B4(CD19)+ IL-2R(CD25)+ PCA-1 +/- TRAP+ . HCLs neither synthesized DNA nor secreted Ig in response to culture with granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin-1 alpha (IL-1 alpha), IL-1 beta, IL-2, IL-3, IL-4, IL-5, or IL-6 . However, a proliferative response (stimulation index greater than or equal to 3.0) without Ig secretion was triggered in HCLs by mitogens or combinations of GFs . Specifically, DNA synthesis was induced at 3 days in three of four HCL samples cultured with Staphylococcus aureus Cowan A (SAC) or the combination of phorbol ester (TPA) and the calcium ionophore A 23187 (Ca2+); DNA synthesis was triggered later (day 7) by tumor necrosis factor (TNF) or by IL-4 and IL-5 . In contrast, the fourth patient, a nonresponder to SAC or TPA/Ca2+, demonstrated increased DNA synthesis at day 3 when cocultured with IL-4 and IL-5 . Both autoradiography and staining with antibromodeoxyuridine (BrdU) MoAb conjugated to fluorescein confirmed DNA synthesis by only a minority (5% to 23%) of tumor cells within each patient . Dual staining confirmed that responsive cells were both BrdU+ and TRAP+ . DNA synthesis induced by TPA/Ca2+ was blocked specifically by anti-IL-6 Ab; in contrast, the HCL proliferative response to SAC, TNF, or IL-4 and IL-5 was not inhibited by anti-IL-6 Ab . alpha-Interferon inhibited the response to TPA/Ca2+, TNF, or IL-4 and IL-5 without any effect on response to SAC . Finally, peroxidase-antiperoxidase staining demonstrated that HCLs are induced by TPA/Ca2+, but not by SAC, to produce intracytoplasmic IL-6 . These data demonstrate IL-4, IL-5, and IL-6 mediated DNA synthesis by HCLs in vitro and suggest a possible in vivo role for these growth factors in the pathophysiology of HCL.

J Immunol, 1990 Nov 15, 145(10), 3372 - 8
Proteolytic fragmentation of sialophorin (CD43) . Localization of the activation-inducing site and examination of the role of sialic acid; Remold-O'Donnell E et al.; Sialophorin (CD43) is the major surface mucin on many hematopoietic cells . It has been implicated in regulating the survival of T lymphocytes in the circulation, and its functions in vitro as the receptor of a T lymphocyte and monocyte activation pathway . The structure of CD43 was examined by protease treatment of lymphoblastoid cells bearing surface CD43 . Trypsin treatment converts CD43 (apparent Mr 115,000) to species of apparent Mr 100,000 called T-100, which remains cell-associated; however, the mechanism of trypsin action was not clarified . Pancreatic elastase and Staphylococcus aureus V8 protease cleave CD43 at discrete extracellular sites . V8 protease generates two fragments, which together account for all properties and mass of the parent molecule . The COOH-terminal fragment V-90 (apparent Mr 90,000) consists of the intracellular and transmembrane regions and part of the extracellular region . The fragment V-30 (apparent Mr 30,000), which is released from the cell, comprises the NH2-terminal approximately 78 amino acids with attached oligosaccharides . V-30 contains the binding sites for the antibodies L2 and L10; the latter is the antibody that activates lymphocytes and monocytes . These findings subdivide the extracellular region of CD43 and indicate that the activation-inducing epitope is located in the most distal portion of the molecule . It is shown that CD43 is insensitive to all but very high concentrations of three proteases . Pretreatment with sialidase enhances sensitivity 13-fold for trypsin, 40-fold for S . aureus V8 protease, and 400-fold for elastase, suggesting that sialic acid influences the survival of surface CD43 molecules when cells are exposed to protease.

Biochem Biophys Res Commun, 1990 Nov 15, 172(3), 1028 - 34
A point mutation in norA gene is responsible for quinolone resistance in Staphylococcus aureus; Ohshita Y et al.; Two norA genes associated with hydrophilic quinolone resistance in Staphylococcus aureus were identified on the two recombinant plasmids pMR8736 and pSA209; the former was derived from a quinolone-resistant strain MR8736, and the latter was derived from a fluoroquinolone-susceptible strain 209P . We compared functions of these two genes, norA8736 and norA209 respectively, by introducing them into E . coli MC1061 . Both genes expressed a novel protein of 52 kilodalton (kD) in size in MC1061 . However, only norA8736 could confer hydrophilic quinolone resistance to the host cell, which was accompanied by a significant decrease in the uptake of a hydrophilic quinolone, norfloxacin, by the cell . Subcloning and recombinant plasmid analyses localized the hydrophilic quinolone-resistance marker to the 0.5 kilobase (kb)-long HpaI-HinfI DNA fragment of pMR8736 . Nucleotide sequencing of this region and the corresponding region of pSA209 revealed that the hydrophilic quinolone resistance conferred by norA8736 was caused by a single nucleotide substitution from A (adenosine) in norA209 to C (cytosine), which corresponded to a single amino acid substitution from Asp to Ala.

J Immunol, 1990 Nov 15, 145(10), 3406 - 11
Leukotriene B4 potentiates the expression and release of Fc epsilon RII/CD23, and proliferation and differentiation of human B lymphocytes induced by IL-4; Dugas B et al.; This study documents the influence of leukotriene (LT) B4 on human B lymphocyte responses . Incubation of freshly isolated B lymphocytes with LTB4, but not LTC4, induced a slight but significant, time- and dose-dependent increase in the surface expression of Fc epsilon RII/CD23 and class II MHC Ag and in the release of soluble CD23 . These changes were maximal at 10 nM LTB4 after an incubation period of 48 h . When B lymphocytes were preactivated in vitro with Staphylococcus aureus Cowan strain I (SAC), neither LTB4 nor LTC4 was able to promote proliferation and/or IgG and IgM secretion . In contrast, when resting B lymphocytes were stimulated with a suboptimal concentration (3 U/ml) of IL-4, LTB4, but not LTC4, potentiated both the Fc epsilon RII/CD23 and the class II MHC antigen expression, and the release of soluble CD23 in a dose-dependent manner, without affecting the kinetics of these responses . Furthermore, LTB4, but not LTC4, amplified both the proliferative response and the IgG and IgM secretion induced by addition of a suboptimal dose of IL-4 (3 U/ml) to SAC-preactivated B lymphocytes . Again, LTB4 did not modify the kinetics of the proliferative response promoted by IL-4 . Although LTB4 potentiated IL-4-induced IgG and IgM secretion from SAC-activated B lymphocytes, no production of IgE was observed . These data indicate that LTB4 could play a regulatory role in the modulation of IL-4-induced signaling in human B lymphocytes.

Eur J Pharmacol, 1990 Nov 13, 190(3), 373 - 9
Mechanisms of pinacidil-induced vasodilatation; Anabuki J et al.; The mechanism of the vasodilator effect of pinacidil was examined . Pinacidil (0.1-100 microM) inhibited the increases in cytosolic Ca2+ ({Ca2+}i) and muscle tension due to norepinephrine in rat aorta . In contrast, a Ca2+ channel blocker, verapamil, inhibited the norepinephrine-stimulated {Ca2+}i more strongly than the contraction . Higher concentrations of pinacidil (3-100 microM) inhibited the verapamil-insensitive portion of the contraction and {Ca2+}i . An inhibitor of ATP-sensitive K+ channels, glibenclamide, antagonized the inhibitory effect of low concentrations (less than or equal to 10 microM) of pinacidol . Pinacidil did not change the contraction induced by Ca2+ in vascular smooth muscle permeabilized with Staphylococcus aureus alpha-toxin . Norepinephrine (in the presence of GTP), 12-deoxyphorbol 13-isobutyrate (in the absence of GTP), and treatment with GTP gamma S potentiated the contraction of permeabilized smooth muscle induced by the addition of Ca2+ . Pinacidil (100 microM) inhibited the potentiation due to GTP gamma S or norepinephrine but not to phorbol ester . These results suggest that pinacidil has dual effects on vascular smooth muscle contraction . At lower concentrations (greater than 0.1 microM), it decreases {Ca2+}i, possibly by activating ATP-sensitive K+ channels . At higher concentrations (greater than 3 microM), it may additionally inhibit the receptor-mediated, GTP-binding protein-coupled phosphatidyl inositol turnover.

Am J Vet Res, 1990 Nov, 51(11), 1826 - 36
Molecular definition of the bovine granulocytopathy syndrome: identification of deficiency of the Mac-1 (CD11b/CD18) glycoprotein; Kehrli ME Jr et al.; Leukocytosis (34,600 WBC/microliter of blood) was detected in an apparently healthy 7-day-old Holstein heifer . Analysis of blood samples obtained over the next 41 days revealed chronic progressive neutrophilia, which peaked at greater than 85% neutrophils and exceeded 100,000 WBC/microliter . In vitro assessment of isolated blood neutrophils obtained from the heifer at 38 and 45 days of age revealed selected functional abnormalities . Endocytosis of immunoglobulin-opsonized Staphylococcus aureus and killing of this test organism by the calf's neutrophils were significantly diminished, as were phagocytosis-associated superoxide generation, chemiluminescence activity, and myeloperoxidase-catalyzed iodination . Diminished H2O2 elaboration by the calf's neutrophils was evident during ingestion of opsonized zymosan or on exposure to phorbol myristate acetate . Extracellular release (secretion) of elastase during ingestion of zymosan was also diminished, although total cell content of elastase was normal, compared with that of neutrophils from age-matched calves, and granular or other morphologic abnormalities of the calf's neutrophils were not evident by ultrastructural examination . Abnormalities of random migration were inconsistently detected, and normal or high degree of antibody-dependent cytotoxicity or natural killing by the calf's neutrophils was observed . Similar in vitro assessment of neutrophils obtained from the calf's dam revealed no functional abnormalities . The calf died at 48 days of age, with persistent fever and chronic diarrhea, despite administration of antibiotics . Histologic examination at necropsy revealed large numbers of intravascular neutrophils in most tissues, including massive neutrophil sequestration in spleen . However, a striking lack of extravascular neutrophils was evident in inflamed submucosa adjacent to intestinal ulcers heavily contaminated with enteric microorganisms . Bone marrow examination revealed diffuse myeloid hyperplasia, but no other abnormalities.

Ned Tijdschr Geneeskd, 1990 Nov 3, 134(44), 2146 - 8
{A patient with toxic shock syndrome following correction of the nasal septum}; Schweitzer DH et al.; A male aged 30 suffered from toxic shock syndrome after septorhinoplasty with positioning of a tampon . Initial treatment consisted of removing the tampon and supportive care, as a result of which the patient recovered . The patient was a carrier of Staphylococcus aureus which produced toxic shock syndrome toxin-I (TSST-I) . Anti-TSST-I antibodies were already found in the serum in the initial phase of the disease.

Int J Pept Protein Res, 1990 Nov, 36(5), 445 - 9
Complete amino acid sequence of a subunit from rapeseed high molecular weight protein; Bhushan R et al.; A subunit (Mr 15,600) from the high molecular weight protein from rapeseed was separated and isolated; its purity and homogeneity were ascertained . The subunit was cleaved with cyanogen bromide, trypsin, chymotrypsin, and Staphylococcus aureus V8 protease . The fragments were separated and isolated by polyacrylamide gel electrophoresis, gel filtration, column chromatography on Dowex 1 x 2, and paper electrophoresis . The amino acid compositions of the intact subunit and different fragments obtained from enzymatic and chemical cleavages were determined . The subunit and its fragments were sequenced by manual Edman method . The phenylthiohydantoin amino acids obtained after each step were identified by thin-layer chromatography and ultraviolet spectroscopy . The complete amino acid sequence of the subunit consisting of 125 amino acid residues has been established by the overlapping method.

DICP . 1990 Nov;24(11):1050, 1053.
Topical vancomycin for the treatment of Staphylococcus epidermidis and methicillin-resistant Staphylococcus aureus conjunctivitis; Ross J et al.; Staphylococcus aureus and Staphylococcus epidermidis are organisms that frequently cause conjunctivitis or blepharoconjunctivitis . We describe a patient with methicillin-resistant S . aureus and S . epidermidis conjunctivitis who was treated successfully using an extemporaneously prepared topical ophthalmic solution of vancomycin hydrochloride 31 mg/mL . Studies describing the preparation, stability, and comfort of this solution, as well as reports pertaining to efficacy, are reviewed . Controlled clinical trials evaluating the safety and efficacy of vancomycin ophthalmic solution have not yet been performed.

Rinsho Byori, 1990 Nov, 38(11), 1211 - 8
{The study of cases with infection by methicillin-resistant Staphylococcus aureus}; Sekikawa T; The purpose of this study was to evaluate the risk factors which lead to death due to highly critical staphylococcic enteritis manifested by high fever, large amount of watery diarrhea and gastric secretion with leukopenia and lymphocytopenia after surgical procedures . We experienced eight cases of severe staphylococcic enteritis by methicillin-resistant Staphylococcus aureus (MRSA) occurred in the early days after operation from 1986 to 1990 . Seven out of eight cases underwent gastrectomy due to gastric cancer . Oral antibiotics were administered in five out of eight cases as the preparation of gastrointestinal tract . Five had been injected the third-generation cephalosporin (CZX) as prophylactic antibiotic administration . In hematological examination, leukopenia and lymphocytopenia were observed in five cases (62.5%) who died soon after operation . So as to prevent the occurrence of MRSA enteritis, it is important to avoid using third-generation cephalosporin after gastrectomy.

Ostomy Wound Manage, 1990 Nov-Dec, 31, 40 - 9
The comparative efficacy and safety of 5% povidone-iodine cream for topical antisepsis; Stahl-Bayliss CM et al.; A 5% povidone-iodine cream (Betadine Cream, The Purdue Frederick Company, Norwalk, Conn.) was tested extensively to determine its safety and efficacy . Results of in vitro microbiologic comparison found that a representative panel of vegetative test organisms couldn't be recovered after 60 seconds or less exposure to povidone-iodine (PVP-I) cream, whereas the kill time of the combination-antibiotic cream (Neosporin Cream, Burroughs Wellcome, Research Triangle Park, NC) exceeded 15 minutes for at least half of vegetative organisms . PVP-I cream produced a log reduction of Bacillus pumilis spores after less than one hour's exposure; the antibiotic cream did not . Both PVP-I and antibiotic creams were essentially non-irritating in human and in vivo animal studies . In open wounds, 5% PVP-I cream caused little or no burning and pain upon application . In human comparisons, artificially induced, standardized lesions inoculated with Staphylococcus aureus, were treated twice daily with PVP-I cream or triple-antibiotic ointment (Neosporin Ointment, Burroughs Wellcome, Research Triangle Park, NC) over three weeks . Both caused significantly reduced bacterial counts (p less than 0.001), and significantly faster healing (p less than 0.05) than no treatment.

Scand J Immunol, 1990 Nov, 32(5), 529 - 36
Alteration of membrane oligosaccharides by castanospermine, an alpha glucosidase inhibitor, enhances immunoglobulin production in Staphylococcus aureus Cowan I-stimulated lymphocyte culture; Karasuno T et al.; Castanospermine (CSP) inhibits alpha-glucosidase, which is involved in the initial step of N-linked oligosaccharide processing of secretory and membrane glycoproteins . In Staphylococcus aureus Cowan I (SAC)-stimulated human lymphocyte culture, CSP at a dose of 20 micrograms/ml caused a twofold increase in immunoglobulin G (IgG) release after 7 days . An initial 48-h exposure to CSP sufficed for this enhancing effect . Plaque-forming cell assays on the seventh day disclosed that CSP caused an increase in the number of IgG-, IgA- and IgM-secreting cells . In cross-culture experiments, only a mixture of B cells pretreated with CSP and untreated T cells showed an increase in IgG production . Tritiated thymidine incorporation studies revealed that CSP enhanced B-cell responses to T cell-derived soluble factor (TSF) . When incubated with CSP for 18 h, B cells showed an increased surface binding on {3H}concanavalin A (Con A) . These results indicate that the alteration in B-cell membrane oligosaccharides enhances the response to TSF at an early stage of SAC culture, leading to an increase in Ig-secreting cell number at later stages . The present study provides evidence that cell-surface oligosaccharides of B cells play an important role in the responses of B cells to lymphokines.

Rev Infect Dis, 1990 Nov-Dec, 12 Suppl 8, S950 - 6
Acute respiratory tract infections among a birth cohort of children from Cali, Colombia, who were studied through 17 months of age; Borrero I et al.; For this study, 340 children less than 18 months old from a low-income, urban neighborhood in Cali, Colombia, were observed from birth by means of weekly home visits to detect cases of acute respiratory tract infection . All suspected cases were confirmed by trained doctors in a special clinic . Information on symptoms, signs, and potential risk factors was documented prospectively . Etiologic agents were identified in cases of lower respiratory tract infection (LRI) . The overall incidence of acute respiratory tract infection was 6.6 cases per child-year at risk . The incidence of upper respiratory tract infection was 4.9 cases per child-year at risk and that of LRI was 1.7 cases per child-year at risk . Crowding in the home was found to be significantly associated with an increased incidence of LRI . Respiratory syncytial virus was the viral agent most frequently isolated from cultures of nasopharyngeal aspirates of children with LRI . Staphylococcus aureus was the bacterial agent most frequently isolated from the blood of patients with LRI.

Appl Environ Microbiol, 1990 Nov, 56(11), 3278 - 84
Sensitive enzyme-amplified electrical immunoassay for protein A-bearing Staphylococcus aureus in foods; Brooks JL et al.; An amperometric electrochemical immunoassay specific for protein A-bearing Staphylococcus aureus was developed . The method was based on a sandwich immunosorbent assay and incorporated an enzyme amplification step, using a NAD-specific redox cycle generating NADH (C . H . Stanley, A . Johannsson, and C . H . Self, J . Immunol . Methods 83:89-95, 1985) . Reduction of the mediator, ferricyanide, was dependent on the initial concentration of antigen . The final potential was measured by using a Pt disk electrode polarized at +0.8 V to the Ag/AgCl reference electrode . The assay was rapid (4 h) and generated protein A- and cell (S . aureus)-dependent signals . The system was highly sensitive and could detect 10 pg of protein A ml-1 and less than 100 CFU of S . aureus ml-1 . Similar sensitivities were observed with S . aureus cultures inoculated into beef and milk, but the sensitivity was reduced slightly (ca . 10(3) g-1) with samples of Cheddar cheese.

Ann Vasc Surg, 1990 Nov, 4(6), 528 - 32
Antibiotic prophylaxis of late bacteremic vascular graft infection in a dog model; Goeau-Brissonniere O et al.; This study was undertaken to determine the efficacy of a single-dose antibiotic injection to prevent late bacteremic vascular graft infection . Twelve dogs had thoracoabdominal aortic bypass with expanded polytetrafluoroethylene grafts . One month later, a bacteremic challenge was produced by rapid intravenous injection of 5 x 10(8) Staphylococcus aureus . Dogs were treated by pairs, each dog of a pair being randomly assigned to receive either 0.5 g ceftriaxone (group I, n = 6) or saline (group II, n = 6), intramuscularly, 90 minutes before challenge . Grafts were harvested seven days after bacteremic challenge . They were cut into 10 fragments, each of which were submitted to bacterial counts . Results of bacterial counts were expressed as colony forming units per square centimeter of graft segment . The overall infection rates were zero of six grafts in group I and four of six in group II (p less than 0.05) . In group I, none of the 60 graft fragments were found to be culture positive (p greater than 0.01) . Bacterial counts from the 24 infected fragments were highly variable, ranging from 12 colony forming units/cm2 to 64 x 10(3) colony forming units/cm2 . Serial quantitative blood cultures revealed a similar decrease of bacteremia in both groups with 2.4 +/- 0.9 x 10(2) (group I) and 1.2 +/- 0.9 x 10(2) (group II) colony forming units/ml at three hours . Mean ceftriaxone serum level was 26 +/- 18 mg/L at the time of bacteremic challenge . These data suggest that a single dose of ceftriaxone given before bacteremic challenge is sufficient to prevent late bacteremic vascular graft infection in this model.(ABSTRACT TRUNCATED AT 250 WORDS)

J Med Microbiol, 1990 Nov, 33(3), 189 - 98
A rat model of Staphylococcus aureus chronic osteomyelitis that provides a suitable system for studying the human infection; Power ME et al.; Chronic osteomyelitis was produced by inoculating Staphylococcus aureus into rat tibia . The infection was characterised grossly by bone deformation and histopathologically by inflammation and the presence of coccal organisms sequestered within the bone tissue . Further observations by scanning electronmicroscopy demonstrated bacteria in microcolonies surrounded by dehydrated amorphous material that was considered to be glycocalyx . Transmission electronmicroscopy, when aided by antibody stabilisation, revealed extensive glycocalyx production within the tibia . These findings indicate that the rat model of chronic S . aureus osteomyelitis mimics the human infection with respect to the sessile mode of growth of bacteria within the bone . Serum antibody levels were assayed by ELISA and immunoblotting procedures . After an initial increase, ELISA titres remained relatively stable, apparently indicating the establishment of chronic osteomyelitis, whereas in immunoblotting an increase in titre over the course of infection was observed . Whole-cell ELISA revealed less subtle differences in antibody titre than did immunoblotting with cell-wall antigen . We found that mid-range antigens, including an antigen implicated as protein A, featured prominently in the immune response in this model of infection.

J Pediatr Orthop, 1990 Nov-Dec, 10(6), 791 - 3
Pyomyositis in an HIV-positive premature infant: case report and review of the literature; Gardiner JS et al.; Pyomyositis is a purulent infection of skeletal muscle caused predominantly by Staphylococcus aureus . Although not often encountered in the continental United States, pyomyositis is frequently seen in tropical areas . Pyomyositis is difficult to diagnose as it may mimic other diseases . Delay in diagnosis may lead to septicemia, shock, and death . Recently, two cases of patients with pyomyositis and acquired immune deficiency syndrome were reported . We report a case of pyomyositis in a 7-week-old premature infant who subsequently tested positive for anti-HIV antibodies . A brief review of the topic is included.

Arthritis Rheum, 1990 Nov, 33(11), 1739 - 44
Outbreak of spontaneous staphylococcal arthritis and osteitis in mice; Bremell T et al.; Staphylococcus aureus is the most common bacterial species found in association with nongonococcal bacterial arthritis in humans . We present here the first description of spontaneous bacterial arthritis and osteitis in mice . Clinically, the most obvious findings were swelling and/or ankylosis of hindpaws and nodose changes of the tail . The prevalence of arthritis and osteitis ranged from 0% to greater than 50% of the mice studied, depending on the mouse strain . The most prominent histopathologic feature of the arthritis was hypertrophy of the synovial tissue and destruction of cartilage and underlying bone . Most of the S aureus-infected mice displayed an identical phage type, which was also the only S aureus phage type found in skin isolates from clinically healthy mice . However, a few S aureus isolates were not typeable, indicating that an additional strain(s) might cause bacterial arthritis in mice.

Ann Thorac Surg, 1990 Nov, 50(5), 800 - 7
Antimicrobial prophylaxis for open heart operations; Miedzinski LJ et al.; Between 1986 and 1988, 450 adults undergoing coronary artery bypass, cardiac valve replacement, or both were enrolled into a prospective, randomized, comparative trial of cephalothin versus cefamandole as perioperative prophylaxis . They were assessed during their hospitalization and at 6 weeks and 6 months after discharge for postoperative infectious complications . Eleven patients had major postoperative infections including 5 with sternal wound infections (three bacteremic), 6 with bacteremia, 1 with prosthetic valve endocarditis, and 3 with severe venous donor graft site infections . Eight major infections occurred in patients receiving cephalothin prophylaxis and three in patients receiving cefamandole, with all five sternal wound infections occurring in the cephalothin group . Postoperative pathogens responsible for the major infections included gram-negative aerobes in 5 patients, Staphylococcus aureus in 4, and Staphylococcus epidermidis in 2 . Preoperative colonizing staphylococcal isolates were not predictive of postoperative staphylococcal pathogens . Although there was no statistically significant difference in rate of major postoperative infectious complications using either cephalothin or cefamandole prophylaxis, there was a trend in favor of cefamandole . Gram-negative aerobes are becoming increasingly important pathogens in this setting.

Am J Vet Res, 1990 Nov, 51(11), 1857 - 9
Detection of capsular polysaccharide in milk of cows with natural intramammary infection caused by Staphylococcus aureus; Sutra L et al.; Detection of capsular polysaccharide (CP) in milk of cows with natural intramammary infection caused by Staphylococcus aureus was attempted . Five quarters of 5 cows harboring S aureus strains that produce type-8 CP were selected . Using an ELISA with a monoclonal antibody, type-8 CP was not detected in extracts prepared from fresh milk collected aseptically . By contrast, CP was easily detectable after incubation of infected milk at 38 C for 20 hours . Quantitation of CP in extracts from incubated milk samples by use of ELISA indicated a great variation of CP expression by strains . Although an incubation step was necessary to detect CP, results of the study indicate that CP may be expressed in vivo during intramammary infection caused by S aureus.

J Trauma, 1990 Nov, 30(11), 1380 - 9
Studies on B-lymphocyte dysfunctions in severely burned patients; Schluter B et al.; We studied in vitro functional parameters of peripheral blood B-lymphocytes from severely burned patients (n = 10; burn injuries ranging from 25 to 72% TBSA) . While the number of B-cells remained unchanged, B-cell proliferation induced by Staphylococcus aureus strain Cowan I (SAC) was normal or even enhanced at early and late phases postburn, but showed a marked suppression during the second to fourth week . A similar pattern was observed for the pokeweed mitogen (PWM)- or SAC-stimulated synthesis of immunoglobulin M (IgM), whereas IgG production was decreased over the whole postburn period monitored . Cytokine (interleukin 4)-induced B-cell activation as indicated by the expression of the CD23 surface antigen was impaired throughout the second to fifth week . In parallel, the release of the proteolytic cleavage product sCD23 which represents a B-cell growth and differentiation factor was reduced . Our data provide evidence that activation, proliferation, and differentiation processes of B-lymphocytes are impaired in severely burned patients, which may contribute to their enhanced susceptibility to infection and sepsis.

J Bacteriol, 1990 Nov, 172(11), 6494 - 8
Characterization of sodium dodecyl sulfate-stable Staphylococcus aureus bacteriolytic enzymes by polyacrylamide gel electrophoresis; Sugai M et al.; Profiles of the bacteriolytic activities of Staphylococcus aureus culture supernatants, sodium dodecyl sulfate cell extracts, LiCl cell extracts, cell wall extracts, and cell membranes were analyzed in sodium dodecyl sulfate-polyacrylamide gels containing Micrococcus luteus or S . aureus . A total of 20 distinct bands of bacteriolytic activity could be detected in gels containing M . luteus, 8 of these bands were found in culture supernatants . The sodium dodecyl sulfate cell extracts, the LiCl cell extracts, and the cell membranes each contained 20 bands (P1 to P20), but no activity was found in cell wall extracts . Less bacteriolytic activity could be detected in gels containing S . aureus, although three bands were found in culture supernatants and LiCl extracts and cell membranes contained one major band, P13 . Crude cell extracts showed five bacteriolytic bands of which the major bacteriolytic bands were distributed in an identical manner in all 10 strains of S . aureus studied . The effects of chemical and physical factors were determined, and it was shown that iodoacetic acid, Hg2+, and Cibacron Blue 3G-A reduced activity, and an optimum pH for enzyme detection was between 7 and 8 . Preincubation at 100 degrees C for 30 min reduced the activity of P1 and P2 bands.

Virology, 1990 Nov, 179(1), 455 - 9
Sequence analysis of the rice dwarf phytoreovirus segment S3 transcript encoding for the major structural core protein of 114 kDa; Suzuki N et al.; The primary structure of rice dwarf phytoreovirus (RDV) genome segment S3 was determined . RDV S3 consists of 3195 nucleotides . A 14-bp segment-specific inverted repeat is located immediately adjacent to the conserved terminal sequence (5'GGCAAA---UGAU3') . A single long open reading frame encoding for 1019 amino acids with an Mr of 114,289 is also identified . In order to investigate the localization of the predicted polypeptide, we determined the amino acid sequence of the 26-kDa peptide fragment obtained from the structural core protein digested by Staphylococcus aureus V8 protease . The sequence of the fragment was found in the translational product presumed from the nucleotide sequence of RDV S3, indicating that RDV S3 encodes the major structural core protein of 114 kDa.

J Orthop Res, 1990 Nov, 8(6), 804 - 13
A model of Staphylococcus aureus bacteremia, septic arthritis, and osteomyelitis in chickens; Daum RS et al.; We studied the occurrence, magnitude, and kinetics of bacteremia and the resultant osteomyelitis and septic arthritis in an avian model of Staphylococcus aureus infection . Thirty-day-old male broiler chicks were inoculated i.v . with 10(5), 10(6), or 10(7) cfu of strain Duntravis, a beta-hemolytic, coagulase-producing, capsular type 8 isolate from the synovial fluid of a 2-year-old black boy . Bacteremia occurred in 80%, 90%, and 100% of animals inoculated with 10(5), 10(6), or 10(7) cfu, respectively . The magnitude of bacteremia in surviving, bacteremic animals increased for 96 hours after inoculation and then decreased after a plateau phase . Osteomyelitis and septic arthritis occurred only in chicks that were continuously bacteremic . The occurrence of osteomyelitis was uniform among continuously bacteremic animals and developed 1 to 23 hours after inoculation . Chickens are susceptible to systemic infections with S . aureus . Bacteremia, osteomyelitis, and septic arthritis may be induced in healthy chickens without prior manipulations that depress their resistance.

Infect Immun, 1990 Nov, 58(11), 3494 - 9
Staphylococcus aureus enterotoxin B challenge of monkeys: correlation of plasma levels of arachidonic acid cascade products with occurrence of illness; Jett M et al.; Arachidonic acid cascade products have been shown to be increased in vitro in Staphylococcus aureus enterotoxin B (SEB)-treated epithelial cell cultures in our laboratory . In order to confirm that these products were clinically related to SEB intoxication, monkeys were administered SEB by nasogastric intubation . It caused emesis in five of six monkeys (less than 4 h), and the sixth monkey showed signs of mild illness . The monkeys which vomited continued to display signs of gastrointestinal illness beyond 8 h but were without any apparent signs of illness by 24 h . Blood samples were collected prior to SEB administration, upon first indication of illness, and at twice that time interval . One week prior to SEB treatment, the same monkeys were administered saline by nasogastric intubation and in every way handled similarly in order to serve as their own controls . Blood samples were taken from the control animals at 0, 4, and 8 h . The plasma concentrations of prostaglandin E2 (PGE2), leukotriene B4 (LTB4), and 5-hydroxyeicosatetraenoic acid (5-HETE) did not vary significantly throughout the 8-h experiment for saline-treated controls, nor did they differ from the concentrations found in the plasma of monkeys just before administration of SEB . When the SEB-treated monkeys showed the first indication of illness (less than 4 h), the mean of the concentration in plasma of PGE2 increased 1.44-fold, that of LTB4 increased 2.23-fold, and that of 5-HETE was essentially unchanged . At twice the time interval of the first display of illness (less than 8 h), PGE2 was still elevated (1.48-fold), LTB4 had decreased slightly to 1.66-fold, and 5-HETE had soared (3,45-fold), suggesting a divergence in the enzymatic utilization of the parent compound of the latter two metabolites, 5-hydroperoxyeicosatetraenoic acid . These studies suggest that arachidonic acid cascade metabolites were a consequence of SEB intoxication and may provide a logical site for metabolic interference in SEB-induced toxicity.

J Immunol, 1990 Nov 1, 145(9), 2974 - 83
A molecular and immunochemical characterization of mouse CR2 . Evidence for a single gene model of mouse complement receptors 1 and 2; Molina H et al.; The relationships between functional, biochemical, and genetic homologues of human and mouse C receptors 1 (CR1) and 2 (CR2) are incompletely understood . We have isolated and characterized a partial mouse CR2 cDNA clone and determined the exon-intron organization of the gene encoding it . Together they predict a form of mouse CR2 highly identical to the 15 short consensus repeat form of human CR2 . Strong similarities in genomic organization and exon-intron junctions indicate that this mouse gene and human CR2 are evolutionary homologues . A polyclonal rabbit anti-mouse CR2 fusion protein, BRN-1, was prepared . BRN-1 immunoprecipitates bands of 155 to 160 kDa under nonreducing conditions in mouse CR2 expressing B cell lines . In mouse spleen a doublet of 155 kDa and 190 kDa under nonreducing and 165 and 205 kDa under reducing conditions is recognized by immunoprecipitation and Western blot analysis . Staphylococcus aureus V8 protease maps of these two proteins show many shared bands . Crossed immunoprecipitation using BRN-1 and 7E9, a previously described mAb reported to identify the 190-kDa mouse CR1 and a smaller 150-kDa protein, indicates that both antibodies react with the same proteins . Therefore, by using BRN-1 we have now linked the genetic mouse CR2 to its functional, biochemically characterized gene product . The observation that BRN-1 also recognizes a second 190-kDa mouse protein defined functionally as a homologue of human CR1, and that these proteins have very similar peptide maps, provides strong evidence that these two proteins are expressed by a single mouse CR2/CR1 transcription unit.

Protein Expr Purif, 1990 Nov, 1(2), 97 - 103
Free-solution isoelectric focusing for the purification of Staphylococcus aureus enterotoxin C1; St Clair NL et al.; A free-solution isoelectric focusing protocol was developed for the preparative purification of Staphylococcus aureus enterotoxin C1 (SEC1) . A toxin consisting of a single isoelectric species, pI 8.8, was purified . Thirty-nine milligrams of SEC1 was recovered from 3 liters of culture supernatant . This significantly improved purification scheme utilized ammonium sulfate precipitation and the Bio-Rad Rotofor isoelectric cell to complete isolation in 2 days, thereby avoiding the protein degradation prevalent when published procedures are used . The purification protocol developed here for SEC1 is used to illustrate the utility of Rotofor fractionation in the general purification of bacterial exotoxins.

Protein Expr Purif, 1990 Nov, 1(2), 104 - 10
Recombinant 70-kDa protein from the amino-terminal region of rat fibronectin inhibits binding of fibronectin to cells and bacteria; Sottile J et al.; Binding of fibronectin to substrate-attached cells and to Staphylococcus aureus is mediated by the amino-terminal 70-kDa portion of fibronectin . The 70-kDa amino-terminus is composed of nine type I and two type II internal homology units, each containing two intrachain disulfide bonds . The exact structural features of the 70-kDa amino-terminus that are necessary for binding to cells and bacteria are not known . We characterized a recombinant 70-kDa protein from the amino-terminus of rat fibronectin using a baculovirus expression system . Recombinant 70-kDa (r70kDa) protein was easily purified in high amounts from the conditioned medium by affinity chromatography on gelatin-agarose . Secretion was much less when N-linked glycosylation was blocked by tunicamycin . Like the native fragment, the r70kDa protein contains intrachain disulfide bonds . In addition, the r70kDa protein was indistinguishable from the nonrecombinant 70-kDa fragment in its ability to compete for binding sites on fibroblasts and S . aureus . Thus, the r70kDa protein retains the important functional characteristics of the native fragment . This expression system is well adapted to studying the structural features important for the interaction of 70-kDa protein with cells.

Enferm Infecc Microbiol Clin, 1990 Nov, 8(9), 572 - 3
{In vitro activity of mupirocin, a new topical agent against Staphylococcus aureus}; Rosales Rodriguez M; The in vitro activity of mupirocin (pseudomonic acid), a new topic antibiotic, was evaluated against 70 S . aureus strains from health care staff of our hospital . The activity of the antibiotic was excellent, with a HIC90 of 0.25 mg/l . Mupirocin activity was higher than that of cloxacillin and bacitracin and similar to that neomycin.

Infection, 1990 Nov-Dec, 18(6), 386 - 7
Brain abscess caused by Brucella abortus and Staphylococcus aureus in a child; Kalelioglu M et al.; A case of a 12-year-old male with Fallot's tetralogy and brain abscess due to Brucella and Staphylococcus is presented . The abscess was aspirated and Brucella abortus and Staphylococcus aureus were isolated.

Anal Biochem, 1990 Nov 1, 190(2), 297 - 303
Biotin-conjugated reagents as site-specific probes of membrane protein structure: application to the study of the human erythrocyte hexose transporter; Deziel MR et al.; A novel labeling procedure using biotin-conjugated protein-modifying reagents has been employed to study the structure and function of the human erythrocyte hexose transporter . The carbohydrate moiety of the isolated, reconstituted transporter was labeled by using galactose oxidase/biotin hydrazide . Cysteine residues, which are essential for transporter function, were tagged with a biotin-conjugated maleimide . Labeling with this reagent inhibited the binding of cytochalasin B to the transporter . Following sodium dodecyl sulfate-gel electrophoresis, labeling of the transporter and its proteolytic fragments was detected by Western blotting and probing with alkaline phosphatase-conjugated avidin . After tryptic cleavage of the transporter into two membrane domains, preparations reacted with galactose oxidase/biotin hydrazide were labeled on the 25-kDa glycosylated fragment, but not on the carbohydrate-free 19-kDa peptide . Biotin-maleimide-labeled cysteine residues on both peptides . Transporter polypeptide was fragmented more extensively using Staphylococcus aureus V8 protease . Limited digestion produced a broad band of 30-50 kDa and sharper bands of 23 and 21 kDa . More extensive digestion resulted in the disappearance of the 23-kDa peptide and the appearance of sharp bands of 20, 19, 17, 13, 11, 8, and 7 kDa . Biotin label introduced with galactose oxidase/biotin hydrazide was found on the broad 30-kDa band, confirming its identity as a glycopeptide . All of the peptides weighing more than 11 kDa contained cysteine residues labeled with biotin maleimide, while the 8- and 7-kDa peptides were unlabeled . These results demonstrate the potential usefulness of biotin-conjugated reagents as site-specific probes of membrane protein structure.

Cytokine, 1990 Nov, 2(6), 464 - 9
Tumor necrosis factor alpha induction in human monocytes; Misuno NI et al.; The present study was undertaken to assess the presence of tumor necrosis factor (TNF)-alpha mRNA and protein in circulating human blood monocytes and to study the TNF-alpha gene expression in human monocytes isolated by continuous Percoll gradient fractionation . The technique of RNA isolation directly from the blood samples was used to study TNF-alpha mRNA expression in circulating human blood leukocytes . It was shown that human blood leukocytes of healthy donors contained no presynthesized pool of TNF-alpha mRNA as well as no TNF-alpha protein . It was found that early pretreatment with cycloheximide interferes with TNF-alpha mRNA induction by Staphylococcus aureus.

Cent Afr J Med, 1990 Nov, 36(11), 278 - 83
Home medication and microbiological profile in chronic otitis media in some Nigerian children; Utsalo SJ et al.; The relationship of home medication to the pathology and microbiological profiles of chronic otitis media was studied in 135 children with middle ear infections . Middle ear lesions were examined and sample obtained for microscopy and culture for bacterial and fungal pathogens . Of a total 115 patients who received home treatment, mostly with extracts of a local herb, Cleome ciliata and unprescribed antibiotics, 111 (96.5 pc) had chronic otitis media with purulent effusion (OME), compared with 11 (55 pc) otitis media (OM) cases among 20 patients who had not received such treatments (p less than 0.05) . Staphylococcus aureus and Pseudomonas spp . were the predominant isolates; the latter were the sole infecting agents in 41 episodes and occurred in 10 co-infections . Sensitivity pattern of bacterial isolates indicated that a large proportion of infections would respond to empiric treatment with gentamicin in anticipation of laboratory results on sensitivity tests . This report suggests further studies to define the significance of these medications in the pathogenesis of chronic OME in Nigerian children.

Antibiot Khimioter, 1990 Nov, 35(11), 21 - 4
{Experimental study on chemotherapeutic efficacy, acute toxic action and nephrotoxicity of combinations of eremomycin with tobramycin}; Shepelevtseva NG et al.; Chemotherapeutic efficacy of eremomycin in combination with tobramycin was investigated on a model of experimental sepsis of albino mice caused by Staphylococcus aureus cultures resistant to methicillin . Eremomycin is a novel original antibiotic of the glycopeptide structure isolated in the USSR and tobramycin is an aminoglycoside . Acute toxicity of the combination with a wide range of the dose fixed proportions was studied on mice and the nephrotoxic action of the antibiotics and their combinations administered intravenously for 5 days was studied on albino rats . The experiments showed that the chemotherapeutic effect of eremomycin in combination with tobramycin was of synergistic nature . Acute toxicity of the combined drugs mainly summed up and somewhat increased when the proportion of tobramycin and eremomycin was 1:2.4 or 1:3.6 . Eremomycin had a dose-depended nephrotoxicity . Summing up of the nephrotoxic action of the drugs on their combined use was observed.

Res Microbiol, 1990 Nov-Dec, 141(9), 1061 - 7
The cellobiose permease of Escherichia coli consists of three proteins and is homologous to the lactose permease of Staphylococcus aureus; Reizer J et al.; The cellobiose (cel) operon of Escherichia coli was recently sequenced and shown to consist of five genes, celABCDF (Parker and Hall, 1990) . We have shown that the CelA, CelB and CelC proteins possess amino acid sequences which are homologous to different domains of the lactose permease of Staphylococcus aureus . CelB corresponds to the integral membrane portion of the permease (IIcel) while CelC (IIIcel) and CelA (IVcel) correspond to the two cytoplasmic domains which appear to comprise the first and second phosphorylation sites in the permease, respectively . The cellobiose permease is the only one of several homologous sequenced permeases of the phosphoenolpyruvate:sugar phosphotransferase system which has its three known functional domains residing on distinct polypeptide chains.

Peptides, 1990 Nov-Dec, 11(6), 1069 - 74
Expression of recombinant human glucose-dependent insulinotropic polypeptide in Escherichia coli by sequence-specific proteolysis of a protein A fusion protein; Chow BK et al.; Glucose-dependent insulinotropic polypeptide (GIP) is a forty-two amino acid hormone that stimulates the secretion of insulin from the pancreatic B-cells in the presence of elevated glucose concentrations . The human GIP gene with the human A alpha-fibrinopeptide sequence was synthesized and linked to the Staphylococcus aureus protein A gene in the vector pRIT2T . This plasmid was expressed in Escherichia coli, and the resulting fusion protein consisted of three domains: protein A for ease of purification, fibrinopeptide sequence for thrombin cleavage and human GIP . The GIP was subsequently cleaved from the fusion protein with alpha-thrombin . The identity of the recombinant human GIP was confirmed by SDS-PAGE, ELISA, HPLC and amino-terminal amino acid sequence analysis . This recombinant product was shown to have comparable insulinotropic activity to porcine GIP in the isolated perfused pancreas.

Biokhimiia, 1990 Nov, 55(11), 2078 - 89
{Hydrolysis of a Staphylococcus aureus cell wall peptidoglycan by 209 P lysoamidase}; Severin AI et al.; Hydrolysis of Staphylococcus aureus 209 P cell wall peptidoglycan was accompanied by the liberation of 1.3 mol of C-terminal and 1.2 mol of N-terminal glycine per mole of Glu as well as of 0.5 mol of N-terminal and 0.3 mol of C-terminal alanine . Gel chromatography on Sephadex G-25, ion-exchange chromatography on QAE-Sephadex A-50 and paper electrophoresis of S . aureus peptidoglycan hydrolysates gave seven homogeneous fractions; these fractions were structurally defined . Lysoamidase hydrolyzed bonds Mur-Ala, Gly-Gly and Mur-GlcN in the peptidoglycan molecule . Hydrolysis of glycan chains was accompanied by the formation of large fragments, (GlcN-Mur)9 and (GlcN-Mur)28 . The lytic effect of lysoamidase on S . aureus peptidoglycan is coupled with bacteriolytic enzymes of lysoamidase: acetmuramyl amidase, glycyl--glycine endopeptidase and acetyl--muramidase.

Zentralbl Bakteriol, 1990 Nov, 274(2), 250 - 8
Effects of beta-lactam antibiotics imipenem/cilastatin and cefodizime on cellular and humoral immune responses in BALB/c-mice; Grochla I et al.; The effects of a 7-day chemotherapy with two broad-spectrum beta-lactam antibiotics (imipenem/cilastatin and cefodizime) on the humoral and cellular immune responses in BALB/c-mice were investigated . Antibiotic dosages were calculated on a body weight basis from therapeutical dosages in human medicine . Treatment of experimental mice with imipenem/cilastatin and cefodizime did not influence the production of immunoglobulines (IgM and IgG) nor the delayed type hypersensitivity to oxazolone . In vitro, exposure of human granulocytes to imipenem/cilastatin and cefodizime did not interfere with their phagocytic activity as determined by chemiluminescence assay . Subinhibitory concentrations of both antibiotics modified Staphylococcus aureus and made them more susceptible for granulocyte phagocytosis in chemiluminescence assays.

Mol Microbiol, 1990 Nov, 4(11), 1957 - 65
Molecular evolution of class A beta-lactamases: phylogeny and patterns of sequence conservation; Pastor N et al.; We present a multiple alignment of the amino acid sequences of eight class A beta-lactamases and utilized it to propose a phylogeny, based on the nucleotide sequences of their corresponding genes . We have also used the alignment, together with the alpha-carbon co-ordinates of the Staphylococcus aureus protein, to search systematically for neighbouring residues that share the same pattern of conservation among the different members of the protein family . The distribution of invariant residues and of groups of residues with co-ordinate changes map, predominantly, at the region of the active site and at interfaces between structural elements, respectively . We have also contrasted the distribution of conserved residues with the positions which are known to differ in mutants and variants of class A beta-lactamases.

Mol Microbiol, 1990 Nov, 4(11), 1947 - 55
Cryptic alpha-toxin gene in toxic shock syndrome and septicaemia strains of Staphylococcus aureus; O'Reilly M et al.; The majority of clinical isolates of Staphylococcus aureus that produce toxic shock syndrome toxin-1 (TSST-1) fail to express alpha-toxin, despite having a copy of the hla gene in the chromosome . The hla gene was cloned from an Hla- TSST-1+ strain, Todd 555, which had been isolated from a case of toxic shock syndrome in the USA . Of the 630 bases of the Todd 555 gene sequenced, 46 differed from the hla gene sequence of strain Wood 46 . The defect in alpha-toxin expression was shown to be due to a nonsense mutation which converted a CAG glutamine codon in the equivalent position in the functional Wood 46 sequence to a TAG stop codon . The same mutation was present in the hla gene cloned from a human septicaemia strain (V37) isolated in Dublin . The nonsense mutation of Todd 555 was suppressed by the supE44 mutation in Escherichia coli resulting in haemolytic activity in cell lysates . Hybrid hla genes were formed by splicing fragments of hla from Todd 555 and Wood 46 . Expression of one such chimaeric hla gene in S . aureus demonstrated that the Todd 555 hla gene has a functional agr-regulated promoter . The silent hla gene may be a cryptic gene in S . aureus.

J Gen Microbiol, 1990 Nov, 136 ( Pt 11), 2231 - 9
Induced deletions within a cluster of resistance genes in the mec region of the chromosome of Staphylococcus aureus; Inglis B et al.; Variants of a methicillin-resistant Staphylococcus aureus showing loss of or reduced resistance to the antibiotic were isolated at frequencies of 0.1-100% from cultures which had been starved, grown at elevated temperature, or given small doses of UV radiation . Three types of variant were identified on the basis of population distribution of resistance to the antibiotic, and field-inversion gel electrophoresis of digests of the chromosome cut with the rare-cutting restriction endonuclease SmaI . Type I variants are methicillin-sensitive and have a deletion in the mec region of the chromosome . Type II variants have reduced methicillin resistance and rearranged DNA elsewhere in the chromosome . Type II variants show reduced methicillin resistance and no detectable change in the chromosome . Type I deletions were mapped using cloned fragments from the mec region . In 13 of the 16 independently isolated deletion mutants, one of the deletion endpoints appears to correlate with the positions of insertion sequences or transposons found in this region of the staphylococcal chromosome.

Nippon Hifuka Gakkai Zasshi, 1990 Nov, 100(12), 1257 - 61
{Combination effects of fosfomycin and other oral antimicrobial agents against methicillin-resistant Staphylococcus aureus}; Akiyama H et al.; Combination effects of fosfomycin (FOM) and other oral antimicrobial agents were studied against methicillin-resistant Staphylococcus aureus (MRSA) (methicillin: minimal inhibitory concentration (MIC) greater than or equal to 12.5 micrograms/ml) isolated from skin and skin structure infections . The fractional inhibitory concentration (FIC) index equal to or less than 0.5 was seen in 63.0% of 27 MRSA strains for FOM and minocycline combination, in 44.4% for FOM and cefatrizine, in 44.4% for FOM and cefaclor, in 40.7% for FOM and cefalexin, in 37.0% for FOM and doxycycline, in 29.6% for FOM and erythromycin, in 22.2% for FOM and rokitamycin, in 18.5% for FOM and ofloxacin, in 14.8% for FOM and sultamicillin, in 11.1% for FOM and clavulanic acid/amoxicillin . The combination effects of FOM and minocyclin, or FOM and cephalosporins were higher than other combinations with FOM . Combination of FOM with other antibiotics could be a useful way to treat MRSA skin and skin structure infections.

Antimicrob Agents Chemother, 1990 Nov, 34(11), 2273 - 6
Cloning and nucleotide sequence of a chromosomally encoded tetracycline resistance determinant, tetA(M), from a pathogenic, methicillin-resistant strain of Staphylococcus aureus; Nesin M et al.; This report describes the cloning and sequencing of a chromosomally encoded tetracycline resistance determinant from a clinical isolate of methicillin-resistant Staphylococcus aureus . On the basis of the sequence, the gene is in the tet(M) class, and it was shown that the S . aureus tetA(M) gene is induced at the level of transcription.

Antimicrob Agents Chemother, 1990 Nov, 34(11), 2260 - 2
In vitro activities of new antimicrobial agents against multiresistant Staphylococcus aureus isolated from septicemic patients during a Belgian national survey from 1983 to 1985; Van der Auwera P et al.; The antimicrobial agents most active against bacteremic isolates of oxacillin-resistant Staphylococcus aureus isolated from 1983 to 1985 were new fluoroquinolones, including PD 117,596 and PD 127,391 (MIC for 90% of isolates {MIC90} in agar, in micrograms per milliliter, 0.1) and temafloxacin, pefloxacin, and ofloxacin (MIC90, 0.4) . Other active antimicrobial agents included fusidic acid (MIC90, 0.2) and fosfomycin (MIC90, 12.5) . Vancomycin was active against all isolates . Mupirocin was very active (MIC90, 0.4).

Antimicrob Agents Chemother, 1990 Nov, 34(11), 2177 - 83
Purification of Staphylococcus aureus beta-lactamases by using sequential cation-exchange and affinity chromatography; Kernodle DS et al.; Boronic acids are active-site inhibitors of serine beta-lactamases, and a phenylboronic acid-agarose affinity column has been used to purify beta-lactamase from crude cell extracts of several bacterial species . We applied phenylboronic acid-agarose chromatography to the purification of Staphylococcus aureus beta-lactamase . Two factors interfered with the success of the previously described single-step chromatographic protocol . First, staphylococcal beta-lactamase exhibited non-active-site-mediated adsorption to the agarose used as a support for the meta-aminophenylborate ligand, preventing the recovery of beta-lactamase from the column . Second, the staphylococcal beta-lactamases exhibited low affinity for meta-aminophenylborate with inhibition constants (Kis) ranging from 8.0 x 10(-3) to 20.0 x 10(-3) M . These problems were resolved by modifying the buffers utilized during chromatography and increasing the dimensions of the affinity column, and a two-stage procedure consisting of cation-exchange chromatography followed by affinity chromatography was used to purify each of the four variants of staphylococcal beta-lactamase . The mean specific activities of the purified type A, B, C, and D beta-lactamases were 44.6, 12.2, 10.6, and 30.8 mumol of nitrocefin hydrolyzed per min/mg of protein, respectively . Dimer formation, presumably from intramolecular cysteine-cysteine cross-linking, was observed with the type D beta-lactamase but not with the type A, B, or C enzyme.

Antimicrob Agents Chemother, 1990 Nov, 34(11), 2122 - 7
In vitro activity of sparfloxacin (CI-978; AT-4140) for clinical Legionella isolates, pharmacokinetics in guinea pigs, and use to treat guinea pigs with L . pneumophila pneumonia; Edelstein PH et al.; The activities of sparfloxacin, ciprofloxacin, and erythromycin for 21 clinical Legionella isolates were determined by agar and broth dilution susceptibility testing and by growth inhibition assays in guinea pig alveolar macrophages (sparfloxacin and ciprofloxacin) . All three antimicrobial agents had roughly equivalent activities when buffered charcoal yeast extract agar medium supplemented with 0.1% alpha-ketoglutarate was used as the test medium; the MICs for 90% of strains were 1.0 micrograms/ml for erythromycin and sparfloxacin and 0.5 microgram/ml for ciprofloxacin . Buffered charcoal yeast extract medium supplemented with 0.1% alpha-ketoglutarate inhibited the activities of all the antimicrobial agents tested, as judged by the susceptibility of a control Staphylococcus aureus strain . Broth macrodilution MICs for two L . pneumophila strains in buffered yeast extract supplemented with 0.1% alpha-ketoglutarate were less than or equal to 0.03 microgram/ml for sparfloxacin, 0.06 microgram/ml for ciprofloxacin, and 0.25 microgram/ml for erythromycin; only erythromycin was inhibited by this medium . Ciprofloxacin and sparfloxacin (both 0.25 microgram/ml) reduced bacterial counts of two L . pneumophila strains grown in guinea pig alveolar macrophages by 2 log10 CFU/ml, but regrowth occurred over a 3-day period . Sparfloxacin, but not ciprofloxacin (both 1 microgram/ml), caused a 3- to 4-day postantibiotic effect . Pharmacokinetic and therapy studies of sparfloxacin were performed in guinea pigs with L . pneumophila pneumonia . For the pharmacokinetic study, sparfloxacin was given (10 mg/kg of body weight) to infected guinea pigs by the intraperitoneal route; peak levels in serum and lung were 2.6 micrograms/ml and 1.6 micrograms/g, respectively, at 1 h, with a terminal-phase half-life of elimination from serum of 5 h.(ABSTRACT TRUNCATED AT 250 WORDS)

Antimicrob Agents Chemother, 1990 Nov, 34(11), 2050 - 4
Ciprofloxacin-resistant methicillin-resistant Staphylococcus aureus in an acute-care hospital; Raviglione MC et al.; Use of ciprofloxacin as an alternative to vancomycin for treatment of methicillin-resistant Staphylococcus aureus infection has been paralleled by the emergence of resistant strains . This phenomenon has also been noticed in our hospital . To confirm our observation, methicillin and ciprofloxacin susceptibilities were tested by disk diffusion and broth microdilution techniques . We studied 83 methicillin-resistant Staphylococcus aureus isolates obtained from various sources over a 4-month period . Ciprofloxacin resistance (MIC, greater than 2 micrograms/ml) was detected in 69 isolates (83%) . Prior use of ciprofloxacin was reported for 24 of 69 patients with ciprofloxacin-resistant strains and 0 of 14 patients with ciprofloxacin-susceptible strains . The day of detection during the hospital stay and the location of the source patient were not significantly different between resistant and susceptible strains . Bacteriophage typing showed a higher occurrence of nontypeable strains among ciprofloxacin-resistant strains (54%) . Review of our microbiology register showed a progressive increase in the rate of resistance to ciprofloxacin during the first year of use, with initial rates being about 10% and recent rates being higher than 80% . On the other hand, methicillin-susceptible S . aureus remained uniformly susceptible to ciprofloxacin (98.4%) . We conclude that prior use of ciprofloxacin is an important factor for the selection of ciprofloxacin-resistant strains and that ciprofloxacin has limited usefulness against methicillin-resistant S . aureus.

J Pharm Pharmacol, 1990 Nov, 42(11), 790 - 4
Cut-off effect in antimicrobial activity and in membrane perturbation efficiency of the homologous series of N,N-dimethylalkylamine oxides; Devinsky F et al.; The antimicrobial activity of the homologous series of N,N-dimethylalkylamine oxides (DMAO) was found to be quasi parabolically dependent on alkyl chain length with a maximum at n approximately 15 and n approximately 12 for Staphylococcus aureus and Escherichia coli, respectively . The physiochemical properties of DMAOs as characterized by critical micelle concentrations, retention times of 1-alkenes generated from DMAOs by gas-liquid chromatography, Rm values in reversed phase chromatography, and bacterial lipid/aqueous phase partition coefficients were found to correlate with the alkyl chain length . The effect of DMAOs on the structure of the model membrane prepared from isolated lipids from Escherichia coli as detected by a spin probe method was maximal for the alkyl chain length n approximately 10-12 coinciding with the maximum in the antimicrobial activity observed with Escherichia coli . It is suggested that the cut-off in the DMAO antimicrobial activity is caused by the cut-off in the DMAO perturbing effect on the membrane structure.

J Formos Med Assoc, 1990 Nov, 89(11), 977 - 81
Opsonizing effect of normal cerebrospinal fluid on Staphylococcus aureus; Hou SC et al.; The opsonizing effect of normal cerebrospinal fluid (CSF) on Staphylococcus aureus has been demonstrated through phagocytic assay . Radiolabelled S . aureus was opsonized using various CSF preparations and then ingested by neutrophils . Uptake of S . aureus by neutrophils was 31 +/- 12% for untreated CSF, 28 +/- 14% for ethylene-glycol-bis(beta-aminoethyl ether) -N,N-tetraacetic acid(EGTA) chelation, 18 +/- 5% for heat-treated (50 degrees C), 14 +/- 8% for heat-inactivated (56 degrees C), 11 +/- 6% for S . aureus-absorbed, and 5 +/- 4% for heat-inactivated and S . aureus-absorbed, respectively . Indirect immunofluorescence antibody assay revealed that IgG was the class of immunoglobulins involved in the opsonic activity of CSF, and complement 3 (C3) deposition was weakly detected in S . aureus . These results suggest that both heat-labile (complement) and heat-stable (antibody) factors are crucial for the opsonization of S . aureus by normal CSF . We conclude that activation of the complement by S . aureus in normal CSF is through an alternative pathway and is enhanced by the presence of a specific antibody (IgG).

Proc Natl Acad Sci U S A, 1990 Nov, 87(22), 8884 - 8
Induction of specific clonal anergy in human T lymphocytes by Staphylococcus aureus enterotoxins; O'Hehir RE et al.; The exotoxins produced by certain strains of Staphylococcus aureus are able to stimulate powerful polyclonal proliferative responses and to induce nonresponsiveness by clonal deletion of T lymphocytes expressing the appropriate T-cell antigen receptor V beta gene products . This paper examines the ability of S . aureus enterotoxins to modulate the responsiveness of human CD4+ T lymphocytes with defined antigen specificity . It was observed that certain S . aureus toxins were able to activate and induce anergy in hemagglutinin-reactive T cells expressing V beta 3+ elements . After exposure to S . aureus enterotoxins A, B, and D in the absence of antigen-presenting cells, the T cells failed to respond to their natural ligand presented in an immunogenic form, despite enhanced proliferation to exogenous interleukin 2 . The S . aureus toxin-induced anergy was associated with modulation of T-cell membrane receptors; down-regulation of the T-cell antigen receptor was concomitant with enhanced expression of CD2 and CD25 . Interestingly, CD28 was increased only on stimulation, suggesting this protein may be differentially expressed by activated and anergic T cells . These results indicate that bacterial toxins are able to induce antigen-specific nonresponsiveness in human T cells, the application of which may be relevant in the regulation of T cells expressing a particular family of V beta gene products.

Antimicrob Agents Chemother, 1990 Nov, 34(11), 2106 - 13
Molecular analysis of a gentamicin resistance transposonlike element on plasmids isolated from North American Staphylococcus aureus strains; Byrne ME et al.; Plasmid-encoded resistance to the aminoglycosides gentamicin (Gm), tobramycin (Tm), and kanamycin (Km) (GmTmKmr) in strains of Staphylococcus aureus isolated in Australia and North America appears to be mediated by one resistance determinant . In Australian isolates, this determinant is flanked by inverted copies of a 1.3-kb insertion sequence, IS256, thereby forming a composite transposon, Tn4001 . Analysis of two conjugative plasmids and a related nonconjugative plasmid from strains of S . aureus isolated in North America showed that the GmTmKmr determinant on these plasmids is also flanked by inverted repeats . In the nonconjugative plasmid, these repeats include only 425 bp of IS256 immediately adjacent to the GmTmKmr region and identical to that on Tn4001 . This truncated Tn4001 element is flanked by copies of the insertion element IS257, and together these elements form a truncated Tn4001-IS257 hybrid transposonlike structure . A third copy of IS257 was located 418 bp from the hybrid structure . The truncated Tn4001 and three repeats of IS257 were present at a conserved site on the plasmids studied . Four additional copies of IS257 were identified on the two conjugative plasmids . These elements flank determinants for resistance to the aminoglycosides neomycin and paromomycin and to ethidium bromide and quaternary ammonium compounds, as well as the region involved in conjugative plasmid transfer.

Antimicrob Agents Chemother, 1990 Nov, 34(11), 2081 - 5
Daptomycin compared with teicoplanin and vancomycin for therapy of experimental Staphylococcus aureus endocarditis; Kaatz GW et al.; The efficacies of daptomycin, teicoplanin, and vancomycin were compared in the therapy of experimental Staphylococcus aureus endocarditis . Rabbits infected with either of two methicillin-susceptible strains (SA-12871 or its moderately teicoplanin-resistant derivative SA-12873) or a methicillin-resistant S . aureus strain (MRSA-494) were treated with daptomycin, 8 mg/kg of body weight, every 8 h; teicoplanin, 12.5 mg/kg (low-dose teicoplanin {teicoplanin-LD}, excluding MRSA-494) or 40 mg/kg (high-dose teicoplanin {teicoplanin-HD}) every 12 h; or vancomycin, 17.5 mg/kg every 6 h, for 4 days . Compared with no treatment daptomycin, teicoplamin-HD, and vancomycin significantly reduced bacterial counts of all test strains in vegetations and renal and splenic tissues (P less than 0.001) . Teicoplanin-LD was equally effective against SA-12871 but failed against SA-12873, with three of six animals still being bacteremic at the end of therapy . For SA-12871, daptomycin was as effective as teicoplanin-HD and was superior to teicoplanin-LD and vancomycin (P = 0.02) in lowering vegetation bacterial counts . There were no differences between daptomycin, teicoplanin-HD, or vancomycin in the reduction of bacterial counts in tissues for any of the test strains . In rabbits infected with SA-12871, vegetations from 33% of teicoplanin-LD-treated, 6% of teicoplanin-HD-treated, and 13% of daptomycin-treated animals yielded organisms for which there were up to eightfold increases in the MICs . Resistance may have contributed to early death in one daptomycin-treated animal . No increases in the MICs for the test strain were detected in animals infected with SA-12873 or MRSA-494 . We conclude that in this model and against these strains of S . aureus, daptomycin and teicoplanin-HD are as efficacious as vancomycin, but diminished susceptibility to both can develop during therapy.

Clin Exp Dermatol, 1990 Nov, 15(6), 415 - 21
Tannic-acid staining material on high endothelial venules and lymphocytes in skin and peripheral lymph nodes in Staphylococcus aureus-associated erythroderma; Heng MC et al.; The recognition and binding of glycoprotein receptors on lymphocytes to specific antigens present on high endothelial venules (HEV) precedes the egress of lymphocytes from the blood stream into the tissues . In this paper, we report the presence of HEVs with tannic-acid staining material (TASM+ HEVs) in Staphylococcus aureus-associated erythroderma, which allow the migration of CD8+ lymphocytes from the bloodstream into the epidermis . TASM positivity is also expressed on lymphocytes within the regional lymph nodes, and by intravascular lymphocytes prior to leaving the TASM+ HEV . It is proposed that TASM positivity may represent a molecule, which may function in binding lymphocytes to HEVs prior to egress from the HEV . (TASM is lost from lymphocytes after leaving the HEVs) . The expression of TASM positivity may form an essential part of the CD8+ lymphocyte-HEV recognition system, and may be the means whereby CD8+ lymphocytes generated in the regional lymph nodes by various mitogens (in this case by staphylococcal mitogens) may 'home' to specific sites within the epidermis . TASM positivity on both the HEVs and lymphocytes may serve as a convenient marker of such a system.

Pacing Clin Electrophysiol, 1990 Nov, 13(11 Pt 1), 1360 - 4
Infections in implantable cardioverter defibrillator patients; Wunderly D et al.; Implantable cardioverter defibrillators (ICDs) have been documented as an effective modality in reducing arrhythmic mortality . A serious complication associated with implantation of the device is infection . Few studies have addressed this issue . Two hundred seven patients with refractory ventricular arrhythmias underwent 207 ICD implantations, and 56 subcutaneous generator changes at our institution . Eight patients developed wound infections, four following ICD implantation (4 out of 207 or 1.9%), and four following a generator change (4 out of 56 or 7.1%) . Wound cultures most commonly revealed Staphylococcus aureus and Staphylococcus epidermidis . Infections treated with antibiotics alone, or with only generator removal, frequently recurred (four out of five attempts) . There were no recurrences following total patch/lead and generator system removal . In five patients, the same generator unit was successfully reimplanted following ethylene oxide sterilization without infection recurrence . We conclude that treatment of device-associated infection generally requires total generator and patch/lead system removal, and that generator units can be successfully reimplanted yielding substantial cost savings.

Biochem J, 1990 Nov 1, 271(3), 635 - 9
Cyclic AMP stimulates luteinizing-hormone (lutropin) exocytosis in permeabilized sheep anterior-pituitary cells . Synergism with protein kinase C and calcium; Macrae MB et al.; Sheep anterior-pituitary cells permeabilized with Staphylococcus aureus alpha-toxin were used to investigate the role of cyclic AMP (cAMP) in exocytosis of luteinizing hormone (lutropin, LH) under conditions where the intracellular free Ca2+ concentration ({Ca2+}free) is clamped by Ca2+ buffers . At resting {Ca2+}free (pCa 7), cAMP rapidly stimulated LH exocytosis (within 5 min) and continued to stimulate exocytosis for at least 30 min . When cAMP breakdown was inhibited by 3-isobutyl-1-methylxanthine (IBMX), the concentration giving half-maximal response (EC50) for cAMP-stimulated exocytosis was 10 microM . cAMP-stimulated exocytosis required millimolar concentrations of MgATP, as has been found with Ca2(+)- and phorbol-ester-stimulated LH exocytosis . cAMP caused a modest enhancement of Ca2(+)-stimulated LH exocytosis by decreasing in the EC50 for Ca2+ from pCa 5.6 to pCa 5.9, but had little effect on the maximal LH response to Ca2+ . Activation of protein kinase C (PKC) with phorbol 12-myristate 13-acetate (PMA) dramatically enhanced cAMP-stimulated LH exocytosis by both increasing the maximal effect 5-7-fold and decreasing the EC50 for cAMP to 3 microM . This synergism between cAMP and PMA was further augmented by increasing the {Ca2+}free . Gonadotropin-releasing hormone (gonadoliberin, GnRH) stimulated cAMP production in intact pituitary cells . Since GnRH stimulation is reported to activate PKC and increase the intracellular {Ca2+}free, our results suggest that a synergistic interaction of the cAMP, PKC and Ca2+ second-messenger systems is of importance in the mechanism of GnRH-stimulated LH exocytosis.

J Immunol, 1990 Nov 1, 145(9), 3054 - 61
Protein L . A bacterial Ig-binding protein that activates human basophils and mast cells; Patella V et al.; Peptostreptococcus magnus strain 312 (10(6) to 10(8)/ml), which synthesizes a protein capable of binding to kappa L chains of human Ig (protein L), stimulated the release of histamine from human basophils in vitro . P . magnus strain 644, which does not synthesize protein L, did not induce histamine secretion . Soluble protein L (3 x 10(-2) to 3 micrograms/ml) induced histamine release from human basophils . The characteristics of the release reaction were similar to those of rabbit IgG anti-Fc fragment of human IgE (anti-IgE): it was Ca2(+)- and temperature-dependent, optimal release occurring at 37 degrees C in the presence of 1.0 mM extracellular Ca2+ . There was an excellent correlation (r = 0.82; p less than 0.001) between the maximal percent histamine release induced by protein L and that induced by anti-IgE, as well as between protein L and protein A from Staphylococcus aureus (r = 0.52; p less than 0.01) . Preincubation of basophils with either protein L or anti-IgE resulted in complete cross-desensitization to a subsequent challenge with the heterologous stimulus . IgE purified from myeloma patients PS and PP (lambda-chains) blocked anti-IgE-induced histamine release but failed to block the histamine releasing activity of protein L . In contrast, IgE purified from myeloma patient ADZ (kappa-chains) blocked both anti-IgE- and protein L-induced releases, whereas human polyclonal IgG selectively blocked protein L-induced secretion . Protein L acted as a complete secretagogue, i.e., it activated basophils to release sulfidopeptide leukotriene C4 as well as histamine . Protein L (10(-1) to 3 micrograms/ml) also induced the release of preformed (histamine) and de novo synthesized mediators (leukotriene C4 and/or PGD2) from mast cells isolated from lung parenchyma and skin tissues . Intradermal injections of protein L (0.01 to 10 micrograms/ml) in nonallergic subjects caused a dose-dependent wheal-and-flare reaction . Protein L activates human basophils and mast cells in vitro and in vivo presumably by interacting with kappa L chains of the IgE isotype.

Appl Microbiol Biotechnol, 1990 Nov, 34(2), 242 - 7
A semi-homogeneous amperometric immunosensor for protein A-bearing Staphylococcus aureus in foods; Mirhabibollahi B et al.; A semi-homogeneous amperometric immunosensor specific to the protein A of Staphylococcus aureus was developed using direct electrochemical detection of phenol produced by alkaline phosphatase from phenyl phosphate . The immunosensor could reliably detect strains of protein A-bearing S . aureus in pure cultures at ca . 10(4) cfu/ml, and at ca . 10(5) cfu/g or ml in various food samples . Due to its semi-homogeneous nature, the system was very simple, easy to operate, and labour-saving . The good correlation between the amperometric current generated by the immunosensor and plate counts illustrated the potential usefulness of this simple system . It proved to be a reliable 24-h detection method for food samples containing very low numbers of protein A-bearing S . aureus after pre-enrichment, as it was able to detect cells that could not directly be enumerated by plate counts.

J Biol Chem, 1990 Oct 25, 265(30), 18414 - 22
An immune complex selective affinity matrix utilizing a synthetic peptide; Baumann MA et al.; A synthetic peptide possessing an amino acid sequence patterned on the globular head region of human complement component 1 subcomponent q (C1q) was tested for immunoglobulin binding . The peptide designated complementary binding peptide 2 (CBP2) was able to inhibit Staphylococcus aureus Protein A and human C1q from binding rabbit immunoglobulin at peptide concentrations for 50% inhibition of 1 and 10 microM, respectively . When attached to a solid-phase matrix in a column, CBP2 was able to bind immune complexes consisting of horseradish peroxidase plus rabbit antiperoxidase antibody or alkali-aggregated human immunoglobulins . A 1:4 mixture of immune complex to free immunoglobulin when passed over the CBP2 column demonstrated selective immune complex binding . Further controls established that CBP2 was in fact binding the immunoglobulin component of the immune complexes in a reversible fashion . The immune complex specificity of the column suggested a functional affinity was forming when CBP2 interacted with immune complexes . The possibility that the sequence of CBP2 is part of the immunoglobulin binding site of human C1q is discussed.

Biochem J, 1990 Oct 15, 271(2), 493 - 9
The smooth muscle 132 kDa cyclic GMP-dependent protein kinase substrate is not myosin light chain kinase or caldesmon; Sarcevic B et al.; Atrial natriuretic peptide (ANP) stimulates the phosphorylation of three cyclic GMP-dependent protein kinase substrate proteins of 225, 132, and 11 kDa (P225, P132 and P11 respectively) in the particulate fraction of cultured rat aortic smooth muscle cells {Sarcevic, Brookes, Martin, Kemp & Robinson (1989) J . Biol . Chem . 264, 20648-20654} . Vrolix, Raeymaekers, Wuytack, Hofmann & Casteels {(1988) Biochem . J . 255, 855-863} have reported the presence of a 130 kDa cyclic GMP-dependent protein kinase substrate protein in the membrane fraction of pig aorta or stomach, and suggested that it may be myosin light chain kinase (MLCK) . The aim of the present study was to determine whether P132 from rat aorta was MLCK or caldesmon . Although P132 co-migrates with purified chicken gizzard MLCK on SDS/polyacrylamide gels, it is distinct from rat aortic MLCK . Partially purified MLCK from rat aorta migrated as a 145 kDa protein on SDS/polyacrylamide gels . Immunoblotting the partially purified rat aortic MLCK with antibody to bovine tracheal MLCK identified rat aortic MLCK (145 kDa) and a corresponding 145 kDa protein in the particulate fraction of cultured rat aortic smooth muscle cells, but did not detect the 132 kDa protein . Phosphopeptide maps of purified rat aortic MLCK prepared by digestion with Staphylococcus aureus V8 protease were distinct from those of P132 . P132 was not caldesmon, since antibodies to caldesmon cross-reacted with 136 and 76 kDa proteins in the particulate fraction of rat aortic cells, but not with P132 . Furthermore, caldesmon was partially extracted from the particulate into the soluble fraction by heating at 90 degrees C, whereas P132 was not . These results demonstrate that the ANP-responsive cyclic GMP-dependent protein kinase substrate of 132 kDa from rat aortic smooth muscle cells is not MLCK or caldesmon.

J Immunol, 1990 Oct 15, 145(8), 2701 - 5
Gene transcription during in vitro activation of human B lymphocytes with Staphylococcus aureus Cowan I strain; Boumpas DT et al.; With the use of nuclear transcription run-off assays we have determined the kinetics of transcriptional activity for the c-fos, c-myc, DR beta, DQ beta, (TfR), Blast-1, and Ig kappa genes in human tonsillar B lymphocytes of high to intermediate density after stimulation with Staphylococcus aureus Cowan I strain (SAC) . Nuclei were isolated at various times after stimulation with SAC . Stimulation with SAC resulted in significant increase in the transcription of all of the genes . Maximum expression of the c-fos, transferrin receptor, DR beta, and DQ beta genes was obtained within 0.5 h after stimulation . Maximum expression of c-myc and Blast-1 was obtained within 3 h . Peak expression of Ig kappa was observed at 8 h . Transcriptional activities of all genes examined, except for the Blast-1 and Ig kappa, decreased rapidly after their respective peaks; they remained at higher levels than their base line . Inhibition of protein synthesis by cycloheximide did not significantly affect the kinetics of transcription of these genes . These data demonstrate the cascade of transcriptional activity of various genes associated with human B cell activation and indicate that the induction of their transcription is independent of de novo protein synthesis.

Blood, 1990 Oct 15, 76(8), 1647 - 56
B-cell differentiation following autologous, conventional, or T-cell depleted bone marrow transplantation: a recapitulation of normal B-cell ontogeny; Small TN et al.; The circulating lymphocytes of 88 consecutive patients following autologous, conventional, or T-cell depleted bone marrow transplantation were serially analyzed for B-cell surface antigen expression and function . In the majority of patients, except for those who developed chronic graft-versus-host disease, the number of circulating CD20+ B cell normalized by the fourth posttransplant month . The earliest detectable B cells normally expressed HLA-DR, CD19, surface immunoglobulin (slg), CD21, Leu-8, and lacked expression of CD10 (CALLA) . In addition, the circulating B cells expressed CD1c, CD38, CD5, and CD23 for the first year following transplant, antigens that are normally expressed on a small percentage of circulating B cells in normal adults, but highly expressed on cord blood B cells . Similar to cord blood B cells, patient B cells isolated during the first year following transplant, proliferated normally to Staphylococcus aureus Cowan strain I (SAC), and produced IgM, but minimal or no IgG when stimulated with pokeweed mitogen and SAC, unlike normal adult B cells that produce both . The similar phenotype and function of posttransplant and cord blood B cells, and their similar rate of decline in patients and normal children adds further evidence to support the hypothesis that B-cell differentiation posttransplant is recapitulating normal B-cell ontogeny.

J Infect Dis, 1990 Oct, 162(4), 967 - 70
Infective endocarditis in intravenous drug users: a comparison of human immunodeficiency virus type 1-negative and -positive patients; Nahass RG et al.; In a study of 34 human immunodeficiency virus (HIV)-seropositive and 12 HIV-seronegative intravenous drug users with 40 and 14 episodes, respectively, of infective endocarditis (IE), there were no differences between groups in the presentation of IE . Staphylococcus aureus was the infecting microorganism in 75% of the HIV-positive patients and 86% of the HIV-negative patients . Overall survival for the HIV-positive patients was 85% compared with 93% for the HIV-negative patients . Ninety percent of patients with asymptomatic HIV infection survived, whereas 60% of patients in CDC group IV survived (P = .052) . In contrast to New Jersey seroprevalence surveys showing HIV antibody in 10%-50% of intravenous drug users, HIV antibody seroprevalence rate in this selected population was 75% . IE in the HIV-seropositive intravenous drug user is essentially the same as in the seronegative user . However, patients with symptomatic HIV infection may be more likely to die from their IE . Intravenous drug users with IE have a greater-than-expected seroprevalence of HIV; the reason remains to be determined.

J Biochem (Tokyo), 1990 Oct, 108(4), 669 - 72
The complete amino acid sequence of a major trypsin inhibitor from seeds of foxtail millet (Setaria italica); Tashiro M et al.; The complete amino acid sequence of a major trypsin inhibitor (FMTI-II) from seeds of foxtail millet (Setaria italica) was determined by analysis of peptides derived from the reduced and S-carboxymethylated protein by digestion with TPCK-trypsin and Staphylococcus aureus V8 protease . FMTI-II consists of 67 amino acid residues, including 10 half-cystine residues which are involved in 5 disulfide bridges in the molecule . The established sequence had a high degree of homology to Bowman-Birk type inhibitors from leguminous and gramineous plants . The trypsin reactive-site peptide bond in FMTI-II also appears to be Lys (16)-Ser (17) by comparison with these sequences.

J Dairy Sci, 1990 Oct, 73(10), 2785 - 9
Shedding pattern of Staphylococcus aureus from bovine intramammary infections; Sears PM et al.; Twenty-one quarters of seven cows were experimentally infected with Staphylococcus aureus (ATCC 29740) to study the shedding pattern in quarter milk samples . Of 991 consecutive quarter milk samples collected from infected quarters during the trial, 745 were positive for S . aureus by bacteriological culture . The sensitivity of a single quarter milk sample to determine infection status of a quarter was 74.5% based on the mean of each gland's recovery pattern . Sensitivity of bacterial culture increased to 94% and 98% by including a second and a third consecutive sample . Because S . aureus is shed in a cyclical manner from mammary glands, consecutive samples would be advisable for accurate diagnosis of infected quarters.

Biull Eksp Biol Med, 1990 Oct, 110(10), 413 - 5
{Antimicrobial effect of irradiation with ionized plasma}; Stupin IV et al.; Ion plasma light flux has been found to possess bacteriostatic effect in vitro and in vivo . Three-fold irradiation with ion plasma of a wound infected by a pathogenic Staphylococcus aureus strain reduced phase I of wound healing by three days and the overall time of healing by four-five days.

Bol Med Hosp Infant Mex, 1990 Oct, 47(10), 698 - 704
{Mucoviscidosis: clinical and laboratory aspects of 29 cases}; Larracilla-Alegre J et al.; The following is a preliminary report on 29 patients with mucoviscidosis . The study was conducted within the hospital and included clinical history, laboratory examinations and X-rays . The patient's ages ranged from four months to 20 years; 15 of them were males, 22 of them began their illness with respiratory symptomatology, 24 of them were previously hospitalized on one or more occasions . Bronchial secretions were examined for pathological and opportunistic bacteria . Pseudomonas and Staphylococcus aureus predominated and were found to be highly resistant to antibiotics . Mortality was high among the group: 13.7%.

J Cell Sci, 1990 Oct, 97 ( Pt 2), 239 - 46
Desmosomal glycoproteins 2 and 3 (desmocollins) show N-terminal similarity to calcium-dependent cell-cell adhesion molecules; Holton JL et al.; The N-terminal sequence of a mixture of desmosomal glycoproteins 2 and 3 (dg2/3, desmocollins) from bovine nasal epidermis, prepared by electro-elution from polyacrylamide gels, was determined by solid-phase Edman degradation . A sequence of 23 amino acids was obtained . This showed 43% identity with that of the N terminus of the calcium-dependent cell adhesion molecule, N-cadherin . A lesser degree of identity with other members of the cadherin-uvomorulin-L-CAM family was also found . In order to confirm that the sequence was derived from the dg2/3 molecules a rabbit antiserum was raised against a synthetic peptide corresponding to the sequence, conjugated to keyhole limpet haemocyanin (KLH) . The antiserum obtained showed high (titre) activity against both the peptide and KLH in ELISA . Each activity could be specifically adsorbed with the appropriate ligand . The antiserum reacted specifically with both dg2 and dg3 of bovine nasal epidermis on immunoblots, this binding was blocked by the N-terminal peptide but was unaffected by KLH . The identity of dg2 and -3 in these preparations was confirmed by immunoblotting with two monoclonal antibodies and one polyclonal antiserum raised against the whole molecules . The N-terminal peptide antiserum was shown to bind to the intercellular space of desmosome profiles by immunoelectron microscopy on ultra-thin frozen sections . One of the two monoclonal antibodies (07-4D) also reacted with the desmosomal intercellular space . dg2 and -3 were shown by Staphylococcus aureus V8 protease digestion to have identical one-dimensional peptide maps . Both the N-terminal antiserum and 07-4D reacted with a V8 fragment of 19,000 Mr derived from dg2 and dg3.(ABSTRACT TRUNCATED AT 250 WORDS)

Food Chem Toxicol, 1990 Oct, 28(10), 687 - 92
Modulation of resistance to mastitis pathogens by pretreatment of mice with T-2 toxin; Cooray R et al.; T-2 toxin, a secondary metabolite of Fusarium species, is a mycotoxin with immunomodulatory activity . In the present investigation the effects of T-2 toxin on host resistance was studied . The virulence of Escherichia coli and Staphylococcus aureus in the mammary glands of mice treated with T-2 toxin was compared with their virulence in control mice . Virulence was estimated from the ability to induce various types of lesions and bacterial growth in the mammary gland . Pretreatment of mice with a single dose (3 mg/kg body weight) of T-2 toxin by gavage reduced the virulence of both E . coli (P less than 0.05) and S . aureus (P less than 0.01) . Microscopic lesions in the infected glands varied in character, from consistently non-reactive necrosis of the entire mammary gland to limited inflammatory reactions . The former were more abundant in control mice than in mice treated with T-2 toxin . Although treatment by gavage with T-2 toxin (0.75 mg/kg body weight/day) for 14 days prior to inoculation had no significant effect on the course of the mastitis infection, virulence was slightly lower in the T-2 toxin treated mice . Both single-dose and successive treatment with T-2 toxin enhanced the respiratory burst activity of macrophages . Pre-inoculation treatment with T-2 toxin also caused a significant increase in the number of peritoneal cells, T-2 toxin did not show bacterial effects on the E . coli or S . aureus strains used for the inoculations . The data indicate that T-2 toxin has modulatory effects on the cell-mediated immune system, and that enhancement of resistance to common mastitis pathogens in mice pretreated with a single dose of T-2 toxin is associated with migration and activation of macrophages.

Indian J Med Res, 1990 Oct, 92, 337 - 40
In vitro effect of intravenous immunoglobulin on serum opsonic activity in normal & intrauterine growth retarded neonates; Kamdar SS et al.; In vitro effect of intravenous immunoglobulin (IVIG), Intraglobin F, on serum opsonic activity against Staphylococcus aureus was studied in 26 full term normal healthy neonates and 18 intrauterine growth retarded (IUGR) neonates by the polymorphonuclear leucocyte overlay method (requiring only a few drops of blood) . Cord IgG and IgM levels were determined by single radial immunodiffusion . Serum opsonic activity against Staph . aureus was significantly lower in the IUGR neonates (49.1 +/- 0.89), as compared to that in normal neonates (61.96 +/- 0.73; P less than 0.001) . Immunoglobulin supplementation in vitro at a concentration of 5 g/dl significantly enhanced the opsonic activity of IUGR neonate sera . Cord IgG levels of IUGR neonates were significantly lower (P less than 0.01) than IgG levels of normal neonates . No significant difference was observed in cord IgM levels between the normal and IUGR neonates.

Biochem Cell Biol, 1990 Oct, 68(10), 1225 - 30
Comparison of D-beta-hydroxybutyrate dehydrogenase from rat liver and brain mitochondria; Zhang WW et al.; The properties of D-beta-hydroxybutyrate dehydrogenase (BDH) from rat liver and brain mitochondria were compared to determine if isozymes of this enzyme exist in these tissues . The BDHs from these tissues behaved similarly during the purification process . The enzymes were indistinguishable by sodium dodecyl sulfate-polyacrylamide or acid-urea-polyacrylamide gel electrophoresis and they had identical isoelectric points . The BDHs from rat liver and brain were also quite similar in functional parameters determined by kinetic analysis and phospholipid activation of apo-BDH (i.e., the lipid-free enzyme) . Antiserum against rat liver BDH inhibited both enzymes to an equivalent extent in a titration assay . The enzymes had similar patterns of peptide mapping by partial digestion with Staphylococcus aureus V8 protease, followed by immunoblotting using antiserum against the liver enzyme . These results suggest that the BDHs in rat liver and brain are very similar and possibly identical.

Nippon Kyobu Geka Gakkai Zasshi, 1990 Oct, 38(10), 2024 - 8
{Mediastinal infection after open cardiac surgery}; Okamoto H et al.; Mediastinal infection is a rare but life-threatening complication after open cardiac surgery . Of 852 patients undergoing cardiac operations performed with a median sternotomy between January of 1981 and August of 1989, 19 patients (2.2%) developed deep sternal infections with mediastinitis, and 6 of them (31.6%) died . Tissue cultures were obtained from all but three patients, and staphylococcus aureus was the most frequent infecting organism associated with high mortality rates (3/8) . 18 patients were managed with debridement and irrigation as the primary treatment, but 4 of them developed infective endocarditis during the course of irrigation treatment and then died . After 1987, 6 patients, who didn't gain wound closure with irrigation alone, were treated with reconstructive techniques, one with pectoral muscle flap closure, another with rectus myocutaneous flap mobilization, and the remainder with omental transfer respectively . All of them achieved eradication of infection as well as wound closure . Then we have advocated early debridement and mobilization of viable tissues such as omentum into the mediastinum.

Nippon Geka Gakkai Zasshi, 1990 Oct, 91(10), 1554 - 9
{Analysis of methicillin-cephem resistant Staphylococcus aureus (MRSA) hospital infection and toxigenicity of MRSA}; Takahashi M et al.; MRSA infectious diseases were often observed in our ward including general, pediatric and neurosurgery during recent two years . Following items were investigated for analysing a prevalence of the infections; 1) the frequency of MRSA in Staphylococcus aureus isolates from clinical materials, 2) the monthly number of patients with MRSA infectious diseases, 3) the biologic types and the toxigenicity of MRSA isolates from clinical materials, nasal carriers and an environmental material in the ward . The results were as follows . 1) Methicillin-cephem resistant strain was determined in 204 of 247 Staphylococcus aureus isolates (83%) . 2) Monthly registration showed a trend that an increased number of the patient in one unit was followed by an increase in other units . 3) Most of MRSA isolates were classified into type II coagulase and two kinds of strains were predominantly found in isolates from clinical materials by plasmid DNA analysis . Most of MRSA isolates had capabilities of producing type C enterotoxin and toxic shock syndrome toxin-1 (TSST-1) . The results suggested that the frequent MRSA infectious diseases attributed to hospital infection . Since hospital infection of virulent MRSA may cause serious infectious diseases, much concern to inhibit the spread of this organism should be required.

Eur J Clin Microbiol Infect Dis, 1990 Oct, 9(10), 717 - 24
Detection of borderline oxacillin-resistant Staphylococcus aureus and differentiation from methicillin-resistant strains; Liu H et al.; Eighty-eight Staphylococcus aureus clinical isolates meeting criteria for borderline oxacillin resistance (intermediate susceptibility or resistance to oxacillin but susceptibility to amoxicillin/clavulanic acid upon disk diffusion testing) were studied to determine optimal test techniques and conditions for differentiating borderline oxacillin-resistant Staphylococcus aureus (BORSA) from methicillin-resistant Staphylococcus aureus (MRSA) . Further testing revealed three distinct resistance patterns: 61 strains (69%) consistently met BORSA criteria and had average beta-lactamase levels five- to six-fold higher than oxacillin-susceptible controls; 11 strains (13%) were markedly heteroresistant MRSA with delayed appearance of resistant colonies leading to spurious susceptibility to amoxicillin/clavulanic acid; 16 strains (18%) appeared to be oxacillin-susceptible on repetitive testing . Under conditions used to elicit intrinsic methicillin resistance in Staphylococcus aureus, a large percentage of BORSA appeared resistant to amoxicillin/clavulanic acid . This clearly shows that BORSA may be misidentified as MRSA while heteroresistant MRSA may appear to be BORSA . It is concluded that amoxicillin/clavulanic acid zone sizes should be measured after a full 24 hours of incubation, that susceptibility testing of Staphylococcus aureus under certain environmental conditions should be interpreted with caution, and that MIC testing is the most reliable technique for differentiating these two resistance patterns in Staphylococcus aureus.

Kansenshogaku Zasshi, 1990 Oct, 64(10), 1275 - 86
{Isolation of methicillin-resistant Staphylococcus aureus (MRSA) at the Hokusho Central Hospital--observation during the recent 4 years}; Mukae H et al.; Clinical and bacteriological studies were carried out to investigate the isolation of MRSA from clinical materials, during the 4 years from 1985 to 1988 at the Hokusho Central Hospital . The isolation frequency of MRSA from Staphylococcus aureus increased from 5.6% (3 strains) in 1985 to 50.0% (90 strains) in 1988 . MRSA from sputum and pressure sore increased significantly . However MRSA from outpatient and inpatient in the ward for common people were constant, about 20% . Inpatients in the ward for aged person were increased significantly from 13.6% (3 strains) to 67.0% (67 strains) . Most of the patients with positive MRSA isolation had background diseases (88.3%) and were bedridden (78.4%) . Patients with cerebrovascular disease were 55.9% of all of the cases . But most of the MRSA strains were resistant to CZX, FOM, DMPPC, CMX, CEZ, CZON, CZX, most of the MRSA strains were sensitive to RFP, VCM, MINO, IPM/CS.

J Antimicrob Chemother, 1990 Oct, 26 Suppl B, 27 - 39
Cellular uptake, localization and activity of fluoroquinolones in uninfected and infected macrophages; Carlier MB et al.; Pefloxacin, like other fluoroquinolones, accumulates in macrophages and several other types of nucleated cells (but not in erythrocytes) . Upon fractionation of macrophage homogenates by isopycnic centrifugation in sucrose gradients, fluoroquinolones are not found associated with any specific cellular structure . We have compared the activities of pefloxacin and roxithromycin against intracellular Staphylococcus aureus in mouse J774 macrophages . Pefloxacin was significantly more active for equivalent intracellular drug concentrations (i.e . expressed by reference to the respective MICs of the drugs as determined in broth), suggesting differences in intracellular availability and/or capacity of the drugs to express their activity in the intracellular environment . The difference was enhanced by incubating the cells in acidic medium . We have also examined the cellular pharmacokinetics and intracellular distribution of pefloxacin in uninfected and Legionella pneumophila infected guinea pig macrophages . In contrast to uninfected cells from which pefloxacin was quickly released, macrophages infected with legionella retained approximately 20-30% of the accumulated pefloxacin after a 60-min wash-out . Cell fractionation studies indicated that the drug remaining in cells was associated with components of high buoyant density . These fractions also contained {3H} if cells had been incubated with {3H} labelled legionella (by in-vitro exposure to {3H}-thymidine, before phagocytosis) . These results suggest that part of the intracellular pefloxacin becomes associated with legionella, or with legionella-containing cytoplasmic structures.

J Rheumatol, 1990 Oct, 17(10), 1364 - 8
Pyogenic sacroiliitis in a rural population; Moyer RA et al.; We describe 10 cases of pyogenic sacroiliitis occurring in a rural population . Seven were male and 3 were female with a mean age of 22.4 years . None was a recent intravenous drug abuser . Five patients had a history of recent pelvic trauma . 99mTechnetium scintiscans revealed increased sacroiliac (SI) joint uptake in 8 of 8 cases . Blood cultures were positive in 60% of patients . Staphylococcus aureus was isolated in 7 cases from blood and/or SI aspirates and Hemophilus influenzae type B in one case . Nine of 10 patients recovered completely . One underwent arthrodesis for recurrent SI pain without evidence of relapse of infection . Median followup was 18 months.

J Antimicrob Chemother, 1990 Oct, 26(4), 567 - 72
Epidemiology of ciprofloxacin resistance among patients with methicillin-resistant Staphylococcus aureus; Smith SM et al.; During the first twelve months after ciprofloxacin was introduced for clinical use at our institution, 65 new patients were found to be either infected or colonized by methicillin-resistant Staphylococcus aureus (MRSA) which were also ciprofloxacin resistant (CR-MRSA) . Only 18 of these patients (28%) had been previously exposed to this antibiotic . Nine (50%) of the 18 patients had received ciprofloxacin for treatment for a pathogen other than MRSA . Although the initial cases of colonization or infection with CR-MRSA can be directly related to ciprofloxacin use, many of the subsequent cases of colonization and infection were not the consequence of ciprofloxacin therapy but rather hospital transmission of existing CR-MRSA.

Can J Vet Res, 1990 Oct, 54(4), 405 - 9
Fragment Bb of bovine complement factor B: stimulatory effect on the microbicidal activity of bovine monocytes; Sethi MS et al.; We have previously shown that the Bb fragment of bovine complement factor B activates bovine monocytes and neutrophils . The activation was demonstrated by the enhanced uptake of 3H-deoxyglucose . To investigate the potential effect of fragment Bb on the microbicidal activity of bovine monocytes, a direct method was used . This method involves an initial ingestion period at 37 degrees C followed by repeated washing . The decrease in the total number of viable intracellular Staphylococcus aureus during the reincubation of the bacteria with bovine monocytes determines the intracellular killing . Maximal intracellular killing was seen when the monocytes containing the ingested S . aureus was incubated with fresh bovine serum (mean +/- SEM = 73.4 +/- 1.4%) . On incubation of the monocytes, containing the ingested bacteria with heat-inactivated bovine serum, 32.5 +/- 0.7% of the intracellular bacteria were killed . When affinity-purified bovine factor Bb was added to the heat-inactivated serum, the intracellular killing capacity was almost restored (65.8 +/- 1.5%) . When monocytes were incubated with medium alone, they killed 22.4% of the intracellular microorganisms . When fragment Bb (25 micrograms/mL) was added to the medium, the intracellular killing of S . aureus doubled (46 +/- 1.29%) . We conclude that the Bb fragment of bovine complement factor B stimulates bovine monocytes in their microbicidal activity.

APMIS, 1990 Oct, 98(10), 881 - 6
Neutrophil chemotactic activity of peptidoglycan . A comparison between Staphylococcus aureus and Staphylococcus epidermidis; Riber U et al.; The ability of peptidoglycan from Staphylococcus epidermidis and Staphylococcus aureus to generate in human serum chemoattractant for peripheral blood neutrophils was studied . It was shown that PG from the two bacteria was able to induce chemotactic activity in normal human serum . Sonication of PG was required to generate this activity . Very little or no activity was generated in heat-treated or C5-deficient human serum by PG, indicating that PG treatment of serum resulted in generation of chemoattractants by activation of complement . Kinetics studies employing C2-deficient or MgEGTA-chelated serum revealed that S . epidermidis induced chemotactic activity by activating the alternative complement pathway . The alternative complement activation induced by S . epidermidis occurred rapidly and was completed after 15 min, whereas S . aureus activated the alternative pathway much more slowly, with activation reaching a maximum at 60 min . The rapid activation of the alternative complement pathway by S . epidermidis PG may partly explain why this bacterium does not normally cause infections in healthy individuals.

Int J Dermatol, 1990 Oct, 29(8), 579 - 82
Staphylococcus aureus in atopic dermatitis and in nonatopic dermatitis; Masenga J et al.; Skin colonization with Staphylococcus aureus (S . aureus) was examined in 30 patients with atopic dermatitis (AD), in 25 patients with nonatopic eczema (NAE) and in 30 individuals as healthy controls (HC) . Bacteria growth was examined in aerobic cultures and the population densities per dish were estimated; S . aureus colonization was found in the eczematous skin of 24 of 30 (80%) AD patients and in 13 of 25 (52%) NAE patients (NS, p greater than 0.1) . In nonaffected skin S . aureus colonization was found in 19 of 30 (63%) of all AD patients compared with 6 of 25 (24%) in NAE patients and 1 of 30 (3%) in HC, respectively (p less than 0.05) . In nonaffected skin, coagulase negative strains of staphylococcus were found in 25 of 30 (84%) controls and in 18 of 25 (72%) NAE patients compared with 12 of 30 (40%) patients with AD . It seems that colonization with S . aureus is not a characteristic feature for atopic dermatitis but is a frequent event in damaged skin; significantly elevated values were also observed in nonatopic eczema . The degree of colonization may depend on the severity and duration of the eczematous lesions.

Arch Dis Child, 1990 Oct, 65(10 Spec No), 1060 - 2
Outcome after acute osteomyelitis in preterm infants; Williamson JB et al.; Eight cases of skeletal infection in preterm infants were studied . All the infants were systemically unwell, with polymorpholeucocytosis . Diagnosis was by blood culture, and any radiographic changes were apparent at the time of presentation . Infection was often multifocal, with sites around the knee being most commonly affected . Staphylococcus aureus was the pathogen isolated in six of the eight cases; in these treatment with fusidic acid was effective and well tolerated, even at doses that were less than the recommended therapeutic minimum . Even with prompt diagnosis and aggressive treatment orthopaedic sequelae are common.

Radiat Res, 1990 Oct, 124(1), 103 - 6
Phagocytic and bactericidal activities of leukocytes in whole blood from atomic bomb survivors; Sasagawa S et al.; This study evaluated the phagocytic and bactericidal activities of peripheral blood leukocytes from Hiroshima and Nagasaki atomic bomb survivors for Staphylococcus aureus . The data were analyzed by multiple linear regression for age, sex, radiation exposure, city of exposure, and neutrophil counts . No significant radiation effect was observed for either blood phagocytic or bactericidal activities . The only significant variable for these functions was the neutrophil count.

J Clin Microbiol, 1990 Oct, 28(10), 2253 - 8
Phagocytosis of mastitis isolates of Staphylococcus aureus and expression of type 5 capsular polysaccharide are influenced by growth in the presence of milk; Sutra L et al.; Phagocytosis by bovine polymorphonuclear granulocytes of seven capsular polysaccharide type 5 Staphylococcus aureus strains isolated from mastitis {corrected} was investigated by means of luminol-dependent chemiluminescence . Bacteria were grown on four different agar media (brain heart infusion, Columbia broth, modified staphylococcus medium 110, and skim milk) and were opsonized by normal bovine serum . When compared to growth on brain heart infusion agar, Columbia agar, and modified staphylococcus medium 110 agar, growth on skim milk agar rendered five of the strains more resistant to opsonization . The other two strains were resistant in all culture media used . Short periods of incubation in milk after growth on brain heart infusion agar did not augment resistance to phagocytosis, indicating that mere adsorption of milk components on bacteria was not responsible . The variability of the chemiluminescence response of polymorphonuclear leukocytes was pronounced among strains with each growth medium except milk . Growth on modified staphylococcus medium 110 and on milk agar favored the masking of teichoic acid, as shown by inagglutinability with rabbit antiserum . Interestingly, agglutination by a monoclonal antibody to capsular polysaccharide type 5 was optimal when bacteria were grown on skim milk agar . This suggests that capsular polysaccharide participated in the masking effect . These findings indicate that masking of the bacterial target of most of the naturally acquired opsonins present in normal bovine serum occurred when bacteria grew in the presence of milk, resulting in an increased resistance to phagocytosis by polymorphonuclear leukocytes.

J Clin Microbiol, 1990 Oct, 28(10), 2161 - 4
Mechanism of plasma clotting by Erysipelothrix rhusiopathiae; Takahashi T et al.; Experiments were performed to determine the mechanism by which Erysipelothrix rhusiopathiae clots plasma . Detection of plasma-clotting activity in four strains of E . rhusiopathiae was carried out by mixing a 24-h broth culture of a tested bacterial strain with rabbit plasma (tube coagulation test) . Sodium citrate, sodium oxalate, EDTA, and heparin were used as anticoagulants in preparing the rabbit plasma . E . rhusiopathiae strains clotted solely citrated plasma in 18 to 24 h . A known coagulase-positive strain of Staphylococcus aureus clotted all of the plasma preparations within 1 h . Various constituents of the organisms, such as cell-free culture filtrates, sonicated extracts, and Formalin-killed bacteria, were also checked for their ability to clot citrated plasma . No constituents of any strain of E . rhusiopathiae clotted the plasma . Only culture filtrates of S . aureus clotted the plasma under these conditions . The spectrophotometric assay demonstrated that two plasma-clotting strains of E . rhusiopathiae consumed the citrate in the plasma just before clotting . Of 301 veterinary clinical isolates of E . rhusiopathiae, 267 (88.7%) were positive in the tube coagulation test . On the basis of these results, it was concluded that plasma clotting by E . rhusiopathiae was due not to extracellular factors such as staphylocoagulase but to consumption of the citrate in the plasma.

J Bone Joint Surg Am, 1990 Oct, 72(9), 1383 - 90
Treatment of infection after total knee arthroplasty by débridement with retention of the components; Schoifet SD et al.; Thirty-one total knee arthroplasties were followed by infection in twenty-seven patients who were subsequently treated with debridement, retention of the components, and intravenous administration of antibiotics . The results were reviewed retrospectively in an effort to evaluate the function of the prostheses that had been salvaged successfully and to identify the causes of failure of those around which an infection had recurred . At the most recent follow-up (average duration, 8.8 years), infection had recurred around twenty-four (77 per cent) of the thirty-one arthroplasties . Seven knees (23 per cent) remained free of infection . Function remained satisfactory, although revision was subsequently needed in two knees for reasons other than infection . One cause of failure was the duration of the infection before debridement . This averaged twenty-one days for the seven knees in which the prosthetic arthroplasty had been salvaged and thirty-six days for the twenty-four knees in which treatment had failed . Another cause of failure was the type of organism: Staphylococcus aureus had caused the infection in fourteen (58 per cent) of the twenty-four knees in which the treatment failed but in only two of the seven knees in which the prosthetic arthroplasty was salvaged . In addition, eight infections in the first group were resistant to penicillin, whereas both infections in the second group were sensitive to penicillin . The four infections with gram-negative organisms in the series were all in knees in which the treatment failed . All six hinged prostheses that were used failed.(ABSTRACT TRUNCATED AT 250 WORDS)

Biochem J, 1990 Oct 1, 271(1), 231 - 6
Determination by photoaffinity labelling of the hydrophobic part of the binding site for acyl-CoA esters on acyl-CoA-binding protein from bovine liver; Hach M et al.; Acyl-CoA esters containing the photoreactive acids 12-(4'-azido-2'-nitrophenoxy){1-14C}dodecanoic acid ({14C}AND-acid) or N-(4'-azido-2'-nitro-{3'-5'-3H}phenyl)-12-aminododecanoic acid ({3H}NANPA-acid) were synthesized . The photoreactive acyl-CoA esters could be bound to bovine acyl-CoA-binding protein (ACBP) and photocrosslinked to the protein . The photocrosslinked acyl-CoA-ACBP complex was separated from unlabelled ACBP on reverse-phase h.p.l.c . and the purified complex was digested with trypsin, Staphylococcus aureus V8 proteinase or endoproteinase Asp-N . By four independent peptide maps it was shown that the amino acids taking part in forming the hydrophobic binding site for acyl-CoA esters in bovine ACBP are located on the peptide segment from Asp21 to Asp38 . Both photoreactive acyl-CoA esters used in this study labelled strongly in the segment from Tyr28 to Ala34 . 12-(4'-Azido-2'-nitrophenoxy){1-14C}-dodecanoyl-CoA ({14C}AND-CoA) also introduced a label at position Asp38, but o labelling was found before Ser29 . In contrast, N-(4'-azido-2'-nitro{3',5'-3H}phenyl)-12-aminododecanoyl-CoA {3H}NANPA-CoA) also labelled the segment from Asp21 to Tyr28 . The difference in labelling by the two photoreactive ligands is most likely caused by different mobility of the arylazido group when linked to the fatty acid either through a phenolic O- or an anilinic N- bond.

Arch Pathol Lab Med, 1990 Oct, 114(10), 1035 - 7
A prospective necropsy study of arthritis in acquired immunodeficiency syndrome; Sokoloff L; Thirty-two knee and 23 sternoclavicular joints from 35 subjects with acquired immunodeficiency syndrome were examined prospectively at necropsy . There were two instances of opportunistic infectious arthritis: one caused by Staphylococcus aureus, the other by Sporothrix schenckii . In five other subjects, para-articular bone was infiltrated by granulomatous or neoplastic sequelae of the human immunodeficiency virus infection . There was no immunohistochemical (p24 antigen) or other evidence for existence of a specific acquired immunodeficiency syndrome arthritis.

Arch Intern Med, 1990 Oct, 150(10), 2151 - 5
Emergence of ciprofloxacin resistance in nosocomial methicillin-resistant Staphylococcus aureus isolates . Resistance during ciprofloxacin plus rifampin therapy for methicillin-resistant S aureus colonization; Peterson LR et al.; We initiated a randomized, single-blinded trial of ciprofloxacin plus rifampin vs sulfamethoxazole and trimethoprim plus rifampin in the therapy for patients who underwent colonization with methicillin-resistant Staphylococcus aureus (MRSA) . Patients who were colonized with MRSA received 2 weeks of either regimen . The study was terminated after the enrollment of 21 subjects due to the recognition of ciprofloxacin resistance in 10 of 21 new MRSA isolates during the last 2 months of the study . Five of the 10 patients with ciprofloxacin-resistant MRSA isolates had never received ciprofloxacin . Long-term (6-month) eradication had been achieved in only three of 11 ciprofloxacin plus rifampin and four of 10 sulfamethoxazole and trimethoprim plus rifampin recipients . The use of this new fluoroquinolone for the eradication of MRSA colonization is usually not effective and may risk the development of ciprofloxacin resistance in MRSA within the hospital environment.

Am Surg, 1990 Oct, 56(10), 648 - 50
Effects of method of hemostasis on wound-infection rate; Ritter EF et al.; Adequate hemostasis is important in preventing postoperative wound infection . This study compared four methods of hemostasis: specific pinpoint vessel electrocautery (SPC), specific vessel ligation with 4-0 vicryl (SVL), nonspecific electrocautery of vessel plus excessive surrounding tissue (NSC), and nonspecific ligation of vessel and excessive surrounding tissue with 4-0 vicryl (NSL), on the rate of wound infection in rabbits that were contaminated with 10(6) Staphylococcus aureus . There was no statistical significant increase in the rate of wound sepsis when electrocautery was used in a fashion producing minimal nonviable tissue compared to specific vessel ligation . Electrocautery use for specific vessel hemostasis does not result in a higher wound infection rate in contaminated wounds.

Am Surg, 1990 Oct, 56(10), 610 - 2
Toxic shock syndrome after inguinal hernia repair . Report of a case with patient survival; Slingluff CL Jr et al.; A 40-year-old man developed fulminant multisystem failure several days after elective repair of an inguinal hernia . Toxic shock syndrome (TSS) was diagnosed . There was, however, no evidence of wound infection at the time of multisystem failure . Only later in his hospital course did the wound drain . Staphylococcus aureus was cultured from the wound and was the presumed etiologic agent in the patient's life-threatening illness . The patient recovered fully with supportive care, antibiotics, and surgical debridement of the inguinal hernia site . This case is discussed in the context of existing literature on the toxic shock syndrome . The site of infection is typically nonsuppurative, but the systemic manifestations are typically life threatening . The responsible organism is commonly believed to be a strain of S . aureus that expresses a toxin (TSS toxin-1) that effects multisystem failure, but which also diminishes the local inflammatory response and explains the benign appearance of the wound . Although this is a rare clinical entity, elective surgical procedures complicated by fatal TSS have been reported . Surgeons should understand this disease and the management necessary to avert mortality.

Epidemiol Infect, 1990 Oct, 105(2), 215 - 28
Outbreaks of infection with methicillin-resistant Staphylococcus aureus on neonatal and burns units of a new hospital; Farrington M et al.; Multiple introductions of methicillin-resistant Staphylococcus aureus (MRSA) strains occurred to a new hospital in Hong Kong . Two years of clinical microbiological surveillance of the resulting outbreaks was combined with laboratory investigation by phage and antibiogram typing, and plasmid profiling . The outbreaks on the special care baby (SCBU) and burns (BU) units were studied in detail, and colonization of staff and contamination of the environment were investigated . MRSA were spread by the hands of staff on the SCBU, where long-term colonization of dermatitis was important, but were probably transmitted on the BU by a combination of the airborne, transient hand-borne and environmental routes . Simple control measures to restrict hand-borne spread on the SCBU were highly effective, but control was not successful on the BU.

Clin Orthop, 1990 Oct, (259), 295 - 303
Results of treatment of tibial and femoral osteomyelitis in adults; Kelly PJ et al.; From January 1, 1971, to December 31, 1985, 425 patients with chronic osteomyelitis of the femur or tibia were seen at the authors' institution . The success rate in this recent experience was 84.4% compared with 50.9% in the authors' results published in 1970 . A classification of chronic osteomyelitis is as follows: (1) hematogenous osteomyelitis; (2) osteomyelitis in united fractures (fracture with union); (3) osteomyelitis in nonunion (fracture with nonunion); and (4) postoperative or posttraumatic osteomyelitis in which bone was not fractured . For management of the scarring of surrounding soft tissue, there has been a change to excision of the scarred tissue and reliance on muscle flap, free-tissue transfer, or closure of soft tissues without irrigation with antibiotic solution . In recent years, free microvascular osseous grafts have permitted more aggressive resection of the involved osseous tissue . The predominance of gram-negative organisms and penicillin-resistant Staphylococcus aureus and the occurrence of methicillin-resistant S . aureus continue.

Clin Exp Immunol, 1990 Oct, 82(1), 181 - 7
Characterization of two monoclonal antibodies (UCL4D12 and UCL3D3) that discriminate between human mantle zone and marginal zone B cells; Smith-Ravin J et al.; Two new monoclonal antibodies (MoAbs), UCL3D3 and UCL4D12 were obtained following immunization with follicular lymphoma (UCL3D3) or low-grade primary B cell gastric lymphoma cells (UCL4D12) . In normal splenic white pulp, tonsil and small intestinal Peyer's patches, UCL4D12 recognizes marginal zone B cells and a subpopulation of follicle centre cells, whereas mantle zone B cells are UCL4D12 negative . In contrast, UCL3D3 recognizes mantle zone B cells and follicular dendritic cells, but not marginal zone B cells or follicle centre B cells . Double-immunofluorescence studies showed that in the splenic white pulp, these antibodies stain reciprocally . The majority of UCL3D3+ cells are sIgM+ and sIgD+ whereas a higher proportion of UCL4D12+ cells express surface IgM (sIgM) but not surface IgD (sIgD) . Less than 10% of splenic B cells express both 3D3 and 4D12 antigens . None of the cell lines tested expressed either antigen . Functional studies showed that both antigens play a role in B cell activation as the MoAbs increase the mitogenic effect of Staphylococcus aureus Cowan I on tonsil B cells . This effect was maximal at 72 h in culture . TPA activation was reduced, and no effect was observed with anti-immunoglobulin (anti mu) or CDw40 (G28.5) . UCL3D3 and UCL4D12 did not show any stimulatory effect on their own . Biochemical studies show that both MoAbs recognize proteins of 80-90 kD under reducing conditions . These two MoAbs appear to recognize new B cell surface antigens which may be useful for identifying subpopulations of B cells.

Arch Biochem Biophys, 1990 Oct, 282(1), 170 - 82
Purification and characterization of cytochrome P450 isozymes from beta-naphthoflavone-induced adult hen liver; Gupta RP et al.; Cytochromes P450 beta NF-A, beta NF-B, and beta NF-C were purified from beta-naphthoflavone-treated adult hens . Cytochrome P450 beta NF-A, however, appeared at two places in the purification scheme . They were designated as cytochromes P450 beta NF-A1 and beta NF-A2 for property comparison . The cytochromes beta NF-A1 and beta NF-A2 were induced by both phenobarbital and beta-naphthoflavone treatment and were similar to P450 PB-A (previously purified from phenobarbital-induced hen livers) in molecular weights, isoelectric pH, spectral properties, behavior on chromatography columns, catalysis of substrates, immunological cross-reactivity on Ouchterlony plates and by immunoblotting, and NH2-terminal amino acid sequence . However, P450 PB-A differed from beta NF-A1/beta NF-A2 in peptide pattern after partial proteolysis by alpha-chymotrypsin and Staphylococcus aureus V8 protease, and complete digestion of 125I-labeled cytochromes by trypsin . The cytochrome P450 PB-A also differed from beta NF-A1/beta NF-A2, in that its antibodies cross-reacted with P-450 of normal, PB-, and beta-NF-induced rabbit liver microsomes . The cytochromes beta NF-B and beta NF-C, although immunochemically cross-reactive with each other, were distinct enzymes on the basis of molecular weights, spectral characteristics, isoelectric pH, peptide pattern on partial proteolysis, tryptic peptide pattern, cross-reactivity of their antibodies with other species, and NH2-terminal amino acid sequence . The most notable difference between beta NF-B and beta NF-C was that the anti-beta NF-C IgG completely inhibited O-dealkylation of 7-methoxyresorufin and 7-ethoxyresorufin by beta-NF-induced microsomes . These activities increased 40- to 50-fold in beta-NF-induced microsomes as compared to only 2- to 4-fold in PB-treated hens . The amino-terminal sequences of beta NF-B and beta NF-C were different from those of mammalian and other nonmammalian species.

J Pediatr, 1990 Oct, 117(4), 531 - 40
Impairment of neutrophil chemotactic and bactericidal function in children infected with human immunodeficiency virus type 1 and partial reversal after in vitro exposure to granulocyte-macrophage colony-stimulating factor; Roilides E et al.; Because polymorphonuclear neutrophils are the most important component of host defense against bacteria, we assessed their function in 13 children with asymptomatic and 12 with symptomatic infection with human immunodeficiency virus type 1 (HIV-1), and compared their values with healthy adult control values . The functions assessed were (1) chemotaxis, (2) bacterial phagocytosis, (3) superoxide generation, and (4) bactericidal activity . Chemotaxis of polymorphonuclear neutrophils toward the chemoattractant N-formylmethionyl leucyl phenylalanine (FMLP) was significantly decreased in symptom-free infected children compared with control subjects (p less than 0.0001), but was increased in children with symptomatic infection (p less than 0.025) . Bactericidal activity of the neutrophils against Staphylococcus aureus was defective in 8 of 12 children with asymptomatic infection (p = 0.016), and in 8 of 9 children with symptomatic infection (p less than 0.00001) . Superoxide generation by polymorphonuclear neutrophils on stimulation with FMLP and phagocytosis of S . aureus were normal . Serum from patients with symptomatic HIV-1 infection was not as efficient in low concentrations as normal serum in the ability to opsonize S . aureus . The in vitro bactericidal defect was partially corrected by granulocyte-macrophage colony-stimulating factor (GM-CSF) . The results suggest that both cellular (neutrophils) and humoral defects contribute to the increased incidence of bacterial infections in HIV-1-infected children, and that GM-CSF may improve the defective bactericidal activity of polymorphonuclear neutrophils in these patients.

J Bacteriol, 1990 Oct, 172(10), 6148 - 50
Chromosomal gene transfer during conjugation by Staphylococcus aureus is mediated by transposon-facilitated mobilization; Stout VG et al.; A chromosomal copy of the transposon Tn551 and a copy coresident on a gentamicin-resistant conjugative plasmid of Staphylococcus aureus resulted in the mobilization of chromosomal genes during filter mating . Gene mobilization was recA dependent and was not restricted to any specific region of the chromosome . Both essential and nonessential genes were transferred.

Infect Immun, 1990 Oct, 58(10), 3307 - 11
Ingestion of Legionella micdadei inhibits human neutrophil function; Donowitz GR et al.; Legionella micdadei is a human pathogen which survives within leukocytes . To determine how this organism escapes intracellular destruction, we examined its effect on human neutrophil activity . Neutrophils were allowed to ingest L . micdadei prior to evaluation of functional activity . Compared with control cells which did not ingest organisms, cells ingesting L . micdadei showed significantly depressed production of superoxide anion (24.5 +/- 9.0 nmol/10(6) cells per 15 min versus 6.9 +/- 3.2 nmol/10(6) cells per 15 min, respectively; P = 0.002), chemotaxis (43.9 +/- 0.8 mm versus 0.9 +/- 1.3 mm of directed migration, respectively; P = 0.001) and bactericidal activity against Staphylococcus aureus (97.9% versus 37.6% of ingested organisms killed, respectively; P = 0.001) . Similar degrees of inhibition could not be demonstrated when either Staphylococcus aureus or Escherichia coli was ingested by cells prior to evaluation . Inhibition of neutrophil function did not occur when phagocytosis of L . micdadei was prevented . However, inhibition occurred with heat-killed as well as with viable organisms . The inhibition of neutrophil function by ingested L . micdadei may help explain the bacterium's ability to survive intracellularly and may begin to explain the pathogenesis of this disease.

Antimicrob Agents Chemother, 1990 Oct, 34(10), 1869 - 74
Antistaphylococcal activities of teicoplanin and vancomycin in vitro and in an experimental infection; Peetermans WE et al.; The efficacies of vancomycin and teicoplanin in an experimental Staphylococcus aureus infection in granulocytopenic mice were related to their activities in vitro and their pharmacokinetic profiles . In vitro teicoplanin had a higher intrinsic activity than vancomycin did; and it also had a more favorable pharmacokinetic profile, resulting in higher peak concentrations in plasma, a longer elimination half-life, and a larger area under the concentration-time curve than those of vancomycin . To predict the antibacterial efficacies of the drugs in vivo on the basis of their activities in vitro and pharmacokinetics, a mathematical model was applied . In the model the in vitro effect was expressed as the difference in growth rate between control cultures and those in the presence of the antibiotic (ER), and the in vivo effect was expressed as the difference between numbers of CFU in control and antibiotic-treated animals (EN) . The integral of ER against time, ERt, was calculated by using the concentrations found in vivo . A significant linear relationship was found between EN and ERt for different dosages at the same times (4 h) after drug administration as well as for the same doses at consecutive times, although at the lowest doses of teicoplanin the observed effect was less than the predicted effect.

J Antimicrob Chemother, 1990 Oct, 26(4), 549 - 59
Assessment of the interaction between ciprofloxacin and teicoplanin in vitro and in neutropenic patients; Lewin CS et al.; The interactions of ciprofloxacin and teicoplanin were investigated against methicillin-sensitive and methicillin-resistant Staphylococcus aureus and Staph . epidermidis isolated during a clinical trial of the efficacy of this combination . In-vitro studies of the combination of ciprofloxacin and teicoplanin found no evidence of any antagonism between these two drugs in terms of inhibition of bacterial multiplication, as determined by Fractional Inhibitory Concentration Indices, and lethality . Clinical use of teicoplanin plus ciprofloxacin as empirical therapy in 29 febrile neutropenic patients revealed an overall response rate of 75% . Response rate for staphylococcal infections, which accounted for 53% of isolated pathogens, was 80% . No serious adverse drug reactions were seen . Our results show that both in vitro, and in the treatment of febrile neutropenic patients, teicoplanin plus ciprofloxacin is an effective anti-staphylococcal combination.

Aust N Z J Surg, 1990 Oct, 60(10), 801 - 3
Establishment of a vascular graft infection model in the sheep carotid artery; Fletcher JP et al.; A study was performed to assess whether a model of vascular graft infection could be established in sheep carotid artery . Either a protein sealed Dacron or a polytetrafluoroethylene (PTFE) graft was used in 22 sheep while a control operation was performed on 4 animals . Staphylococcus aureus in concentrations of 10(2), 10(4), 10(6) or 10(8) colony-forming units (CFU) was inoculated into the wound before closure . No infection occurred with an inoculum of 10(2) . The lowest concentration of organism producing infection was 10(4) for PTFE, 10(6) for Dacron and 10(8) for controls . Six of eleven Dacron grafts and seven of ten PTFE grafts became infected . The sheep carotid artery has proven a satisfactory model for studies of vascular graft infection.

J Neurochem, 1990 Oct, 55(4), 1442 - 5
Mutation of serine 41 in the neuron-specific protein B-50 (GAP-43) prohibits phosphorylation by protein kinase C; Nielander HB et al.; The neuron-specific, calmodulin-binding protein B-50 (also known as GAP-43, F1, or neuromodulin) is an endogenous substrate of protein kinase C (PKC) . PKC exclusively phosphorylates Ser residues in B-50 . As potential phosphorylation sites for PKC, Ser41, Ser110, and Ser122 were indicated, of which Ser41 is contained in the sequence ASF, which matches with the sequence of a synthetic PKC substrate . N-terminally 35S-labeled B-50, produced from cDNA, was subjected to digestion with Staphylococcus aureus V8 protease (SAP) . Consecutively, 35S-labeled 28- and 15-kDa fragments were formed, similar to those after digestion of 32P-labeled B-50 . In a previous study, we showed that the 32P-labeled 15-kDa SAP fragment contains all 32P radioactivity . The present data indicate that it contains the N-terminus of B-50 as well . The 15-kDa fragment, with a calculated length ranging from amino acid residue 1 to 65, contains only one potential PKC phosphorylation site, at Ser41 . Mutagenesis of Ser41 into Thr or Ala resulted in recombinant B-50 products with mobilities on two-dimensional electrophoresis similar to those of the nonmutated recombinant B-50 and the rat brain B-50 . Only {Ser41}B-50 was phosphorylated by PKC, whereas {Thr41}- or {Ala41}B-50 did not show any phosphorylation at the positions indicated on the immunoblots . This leads us to the conclusion that Ser41 is the sole phosphorylation site for PKC in vitro.

Blood Coagul Fibrinolysis, 1990 Oct, 1(4-5), 375 - 84
Expression in Escherichia coli of a recombinant fragment (Ile 914-Leu 1364) of human von Willebrand factor containing a collagen binding domain; Jorieux S et al.; Previous studies have shown that a collagen binding domain of human von Willebrand factor (vWF) resides on a fragment named SpI obtained by digestion with Staphylococcus aureus V8 protease which corresponds to residues Gly 911-Glu 1365 of the mature plasma vWF subunit . We have subcloned a fragment of a full-length cDNA encoding vWF into an expression vector which uses an inducible lambda PL promoter . The predicted product expressed by this plasmid is a fusion protein consisting of 16 amino acids (aa) of the lambda cII protein and aa Ile 914-Leu 1364 of human vWF . This fusion protein was shown to be expressed as insoluble inclusion bodies by induced E . coli harbouring the recombinant vector and was partially purified from bacterial debris and renatured . Partially purified bacterial extract run on SDS-polyacrylamide gels contained a major band representing 50-70% of the visualized proteins and corresponded to the predicted fusion protein . This band reacted with both polyclonal antibodies against human vWF and monoclonal antibodies (MAbs) which recognize the SpI fragment of plasma vWF . The radiolabelled partially purified bacterial extract was shown to bind specifically to human fibrillar collagen types I and III . This binding, which was a function of radiolabelled ligand and collagen concentrations, did not occur on monomeric denatured collagen . It was inhibited by both unlabelled bacterial extract and plasma vWF and also by a MAb against vWF SpI, which has the property of inhibiting vWF/collagen interaction . Our data demonstrate that this recombinant vWF SpI fragment, which can be obtained in large amounts, is a useful model for localizing the epitopes of monoclonal antibodies and then for studying the mechanism of vWF/collagen interaction.

Indian J Pathol Microbiol, 1990 Oct, 33(4), 299 - 303
Changing phage pattern of Staphylococcus in pyoderma cases; Baslas RG et al.; Staphylococcus aureus is a major pathogen for pyoderma in India . Phage pattern of bacteria gives valuable information in epidemiological studies of infection . Two hundred and two strains of Staphylococcus aureus isolated from pyoderma cases at Gorakhpur, were phage grouped and phage typed . It was found that 43.1 percent strains were not typable . Most common group was mixed phage group (23.8 percent) followed by phage group III (12.4 percent) . Predominant phage types in mixed phage group was 84/81/85 and in phage group III was 84/85.

Microb Pathog, 1990 Oct, 9(4), 235 - 41
Respiratory syncytial virus modulation of adult and neonatal lymphocyte mitogenic responses and the role of interferon-gamma; Paton AW et al.; Infection of human adult and neonatal mononuclear leukocytes (MNLs) with respiratory syncytial virus (RSV) caused significant inhibition of lymphoproliferation in response to various mitogens (heat-killed Staphylococcus aureus, phytohaemagglutinin, concanavalin-A and pokeweed mitogen) . Antibody to human interferon-gamma enhanced the RSV-induced suppression of lymphoproliferation in adult, but not neonatal MNLs . These results are consistent with the hypothesis that RSV can suppress the development of a cell-mediated immune response by direct interaction with lymphocytes, and at least in adult cells, interferon-gamma could be implicated in resistance to this action.

J Biochem (Tokyo), 1990 Oct, 108(4), 544 - 8
Purification and characterization of two forms of fatty acid omega-hydroxylase cytochrome P-450 from rabbit kidney cortex microsomes; Yoshimura R et al.; We have previously reported the isolation of two forms of cytochrome P-450 (P-450) with omega-hydroxylase activities toward prostaglandin A (PGA) and fatty acids, designated as P-450ka-1 and P-450ka-2, from kidney cortex microsomes of rabbits treated with di(2-ethylhexyl)phthalate {Kusunose, E . et al . (1989) J . Biochem . 106, 194-196} . In the present work, we have purified and characterized two additional forms of rabbit kidney fatty acid omega-hydroxylase, designated as P-450kc and P-450kd . The purified P-450kc and P-450kd had specific contents of 13 and 16 nmol of P-450/mg of protein, with apparent molecular weights of 52,000 and 55,000 on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), respectively . Both the forms showed absorption maxima at 450 nm in the carbon monoxide-difference spectra for their reduced forms . These P-450s efficiently catalyzed the omega- and (omega-1)-hydroxylation of fatty acids such as caprate, laurate, myristate, and palmitate, in a reconstituted system containing P-450, NADPH-P-450 reductase, and phosphatidylcholine . Cytochrome b5 stimulated the reactions to only a slight extent . They had no detectable activity toward PGA and several xenobiotics tested . The two P-450s showed different peptide map patterns after limited proteolysis with papain or Staphylococcus aureus V8 protease.(ABSTRACT TRUNCATED AT 250 WORDS)

Methods Find Exp Clin Pharmacol, 1990 Oct, 12(8), 541 - 4
In vitro antimicrobial susceptibilities of 349 methicillin-resistant Staphylococcus aureus isolates from veterans; Flournoy DJ et al.; Three hundred and forty-nine methicillin resistant Staphylococcus aureus (MRSA) isolates from veterans were tested (by disc agar diffusion) for their in vitro activity against 18 antimicrobial agents . At least 90% of the isolates were susceptible to bacitracin, nitrofurantoin, hydrogen peroxide, novobiocin, netilmicin and vancomycin . We feel that the aminoglycoside, netilmicin, might provide an alternative agent (to intravenously administered vancomycin) for treating multiply-antimicrobial resistant MRSA . In addition, hydrogen peroxide exhibited very good activity against the test isolates and may have some use as a topical agent for reduction of MRSA on skin and some mucous membranes . This study suggests that further evaluation of netilmicin and hydrogen peroxide (topical only) might be useful.

Electrophoresis, 1990 Oct, 11(10), 883 - 91
Development of a database of amino acid sequences for human colon carcinoma proteins separated by two-dimensional polyacrylamide gel electrophoresis; Ward LD et al.; The tandem use of preparative two-dimensional polyacrylamide gel electrophoresis (2-DE) and electroblotting onto polyvinylidene difluoride membranes has been employed to rapidly isolate a number of proteins from a crude cell extract of a human colon carcinoma cell line (LIM 1863) . The immobilized proteins were located by staining with Coomassie Brilliant Blue R-250, and selected protein spots were excised and subjected to Edman degradation . Our results demonstrate that overall sequence yields in the 3-20 pmol range can be achieved on protein spots from four identical 2-DE gels; approximately 150-200 micrograms of total protein was applied to a single 2-DE gel . An approximate two-fold increase in sensitivity of phenylthiohydantoin-amino acid detection (subpicomole range) was achieved by fitting our commercial sequencers with a simple sample transfer device which permitted the analysis of the total phenylthiohydantoin-amino acid derivative . N-Terminal amino acid sequence data was obtained for thirteen electroblotted proteins . All of these sequences positively matched those of proteins of known structure listed in the available protein sequence databases . Approximately 40% of the electroblotted proteins did not yield N-terminal sequence information, presumably because they had blocked N-termini (either naturally or artifactually) . Internal amino acid sequence information was obtained from three proteins isolated by preparative 2-DE . This was achieved by in situ digestion of the proteins in the gel matrix with Staphylococcus aureus V8 protease, electrophoresis of the generated peptides in a one-dimensional gel, electrotransfer of the peptides to a polyvinylidene difluoride membrane and microsequence analysis of the electroblotted peptides.

J Formos Med Assoc, 1990 Oct, 89(10), 896 - 900
Staphylococcus aureus endocarditis in drug addicts: report of 2 cases; Chen YC et al.; Two young men presented with prolonged hectic fever and chills followed by chest pain, dyspnea and hemoptysis . The chest films revealed multiple lung infiltrates, and blood cultures yielded Staphylococcus aureus . Echocardiographic examination confirmed the diagnosis of tricuspid valve endocarditis . Multiple punctate lesions in the bilateral inguinal areas and dragon tattoos over the forechest gave rise to the suspicion of drug abuse . After prolonged antimicrobial therapy, bacteremia was eliminated, and elective vegetectomy and valvuloplasty were performed on one of the patients . The other one suffered recurrent episodes of pulmonary embolism . Disappearance of the large vegetation was disclosed by echocardiography . Both of them eventually regained their health with the abstinence of drugs . This report illustrates two typical cases of infective endocarditis in drug addicts.

J Hosp Infect, 1990 Oct, 16(3), 231 - 9
The impact of methicillin- and aminoglycoside-resistant Staphylococcus aureus on the pattern of hospital-acquired infection in an acute hospital; Meers PD et al.; Infections due to methicillin- and aminoglycoside-resistant Staphylococcus aureus (MARSA) appeared in a new teaching hospital shortly after it opened . The effect this had on the pattern of hospital-acquired infections in the four years that followed is described . No control measures were applied and MARSA became endemic . New infections appeared at a rate of about four for each 1000 patients discharged . It established itself at different levels of incidence in various specialist units, patients under intensive care being most severely affected . MARSA was implicated in half of all hospital-acquired infections due to S . aureus but it was not more pathogenic than its more sensitive counterpart . It had little impact on the life of the hospital.

J Bacteriol, 1990 Oct, 172(10), 5783 - 8
Cloning and expression of a Staphylococcus aureus gene encoding a peptidoglycan hydrolase activity; Jayaswal RK et al.; A gene of Staphylococcus aureus PS47 encoding lytic activity was cloned and expressed in Escherichia coli . Deletion analysis of a recombinant plasmid carrying a 7.4-kilobase-pair fragment (kbp) of S . aureus DNA suggested that the gene was located within a 2.5-kbp EcoRI-XbaI fragment . Analysis of extracts of E . coli harboring recombinant plasmids on denaturing polyacrylamide gels containing purified cell walls of S . aureus showed a clearing zone by a polypeptide of apparent Mr 23,000 . The release of dinitrophenylalanine but not reducing groups from purified cell walls by a cell extract of recombinant E . coli suggested that we had cloned an N-acetylmuramyl-L-alanine amidase.

Antimicrob Agents Chemother, 1990 Oct, 34(10), 1925 - 31
Assessment of effects of protein binding on daptomycin and vancomycin killing of Staphylococcus aureus by using an in vitro pharmacodynamic model; Garrison MW et al.; Initial clinical trials with daptomycin (2 mg/kg per day) were prematurely suspended because of unexplained treatment failures in patients with bacteremia who were treated with daptomycin, despite in vitro data indicating that the gram-positive cocci causing the infection were susceptible to daptomycin . One explanation for these clinical failures may relate to the relatively high degree of daptomycin protein binding (94%) . To evaluate the impact of protein on daptomycin activity, a two-chamber in vitro pharmacodynamic model was used to study and compare the interaction between Staphylococcus aureus (clinical isolate) and either daptomycin or vancomycin, each in the presence and absence of physiologic human albumin concentrations . Low-dose (2 mg/kg) daptomycin, high-dose (6 mg/kg) daptomycin, and 10 mg of vancomycin per kg beta-phase elimination serum-concentration-versus-time curves were simulated by using this in vitro pharmacodynamic model . The bacterial kill rates by all three regimens were decreased in the presence of albumin (P less than 0.0002) . The average times required for a 99% kill of the initial S . aureus inocula (approximately 5 x 10(7) CFU/ml) without albumin were 0.81 (low-dose daptomycin), 0.33 (high-dose daptomycin), and 6.18 (vancomycin) h . The average times required for a 99% kill of S . aureus with albumin were 7.66 (low-dose daptomycin), 0.95 (high-dose daptomycin), and 10.52 (vancomycin) h . These data demonstrate that, depending on the concentration of daptomycin, the presence of albumin can profoundly diminish the bactericidal activity of daptomycin.

Chem Pharm Bull (Tokyo), 1990 Oct, 38(10), 2880 - 1
Mode of action of the gramicidin S analogs lacking hydrophilic amino acid residues on biomembranes; Katsu T et al.; The gramicidin S analog lacking basic ornithine residues, cyclo(-Val-Ala-Leu-delta Phe-Pro-)2 (where delta Phe represents alpha, beta-dehydrophenylalanine), increased the K+ permeability of human erythrocytes and Staphylococcus aureus similarly to the parent gramicidin S . This analog altered the normal discoid shape of human erythrocytes to an invaginated form . The direction of the shape change was opposite to the case of gramicidin S causing crenated cells . We suppose that the analog accumulated predominantly into the inner half monolayer of membrane and destabilized the membrane structure, resulting in a break in the membrane.

J Med Microbiol, 1990 Oct, 33(2), 91 - 6
Application of small fragment restriction endonuclease analysis (SF-REA) to the epidemiological fingerprinting of Staphylococcus aureus; Haertl R et al.; Total cell DNA of 14 isolates of Staphylococcus aureus from patients of an intensive care unit (ICU) and 180 unrelated strains was examined by restriction endonuclease analysis (REA) . EcoRI-generated DNA fragments were either subjected to conventional REA on agarose gels and stained with ethidium bromide or separated by polyacrylamide gel electrophoresis and visualised by silver staining (SF-REA) . Both methods were compared for inter-strain discriminatory ability, reproducibility and handling . All DNA-cleavage patterns of unrelated strains clearly differed from each other when subjected to SF-REA . In contrast, all S . aureus isolates from the ICU gave identical restriction fragment patterns . These findings supported the suspicion of nosocomial infection in these patients . Conventional REA proved the identity of the ICU isolates, but it failed to differentiate between some of the unrelated strains . Therefore SF-REA of total cell DNA seemed to be superior . It has proved to be a very useful technique for studying the epidemiology of S . aureus in hospitals.

FEBS Lett, 1990 Oct 1, 271(1-2), 131 - 6
Importance of the alpha 3-fragment of complement C4 for the binding with C4b-binding protein; Hessing M et al.; The human regulatory complement component C4b-binding protein (C4BP) is a multimeric plasma protein, which regulates the classical pathway of the complement system . C4BP functions as a cofactor to factor 1 in the degradation of C4b and accelerates the decay rate of the C4b2a complex . Previously, we have demonstrated that monoclonal antibodies (C4-2 and 9) directed against the alpha'-chain of C4b inhibit the binding of C4b to C4BP . In order to identify the structural domain of C4b that binds C4BP, proteolytic fragments of C4 were generated with trypsin and Staphylococcus aureus V8 protease . Sodium dodecyl sulfate polyacrylamide gel electrophoresis, immunoblotting and amino acid sequence analysis of the proteolytic fragments reactive with the anti-C4 mAb's revealed that the residues Ala738-Arg826 of the alpha 3-fragment of C4b are important for the interaction with C4BP.

Gene, 1990 Sep 28, 94(1), 37 - 43
Cloning, expression and characterization of the Sau3AI restriction and modification genes in Staphylococcus carnosus TM300; Seeber S et al.; The genes encoding the restriction enzyme (ENase) and modification enzyme (MTase) of Staphylococcus aureus 3A (recognition sequence 5'-GATC-3') have been cloned in Staphylococcus carnosus TM300 using the vector pCA44 . Clones carrying both genes were isolated from DNA libraries prepared with MboI + BamHI . The DNA region encoding M.Sau3AI was subcloned on a 3.66-kb EcoRV fragment in vector pT181mcs . Plasmids purified from the clones were resistant to digestion with Sau3AI, indicating that the sau3AIM gene was expressed and the product was functional in S . carnosus . Cell lysates of clones with both activities encoded on plasmid pSEM7, cut DNA with the same pattern as Sau3AI, showing that the sau3AIR gene was also expressed and the ENase was functional in S . carnosus . Sequence analysis shows that both genes are transcribed in the same direction and encode polypeptides with calculated Mrs of 56,477 for R.Sau3AI and 47,300 for M.Sau3AI . Efforts to clone one or both genes in Escherichia coli have so far failed.

Biochem J, 1990 Sep 15, 270(3), 651 - 7
The amino acid sequence of cytosolic aspartate aminotransferase from human liver; Doyle JM et al.; 1 . The cytosolic aspartate aminotransferase was purified from human liver . 2 . The isoenzyme contains four cysteine residues, only one of which reacts with 5,5'-dithiobis-(2-nitrobenzoic acid) in the absence of denaturing agents . 3 . The amino acid sequence of the isoenzyme is reported, as determined from peptides produced by digestion with trypsin and with CNBr, and from sub-digestion of some of these peptides with Staphylococcus aureus V8 proteinase . 4 . The isoenzyme shares 48% identity of amino acid sequence with the mitochondrial form from human heart . 5 . Comparisons of the amino acid sequences of all known mammalian cytosolic aspartate aminotransferases and of the same set of mitochondrial isoenzymes are reported . The results indicate that the cytosolic isoenzymes have evolved at about 1.3 times the rate of the mitochondrial forms . 6 . The time elapsed since the cytosolic and mitochondrial isoenzymes diverged from a common ancestral protein is estimated to be 860 x 10(6) years . 7 . Experimental details and confirmatory data for the results presented here are given in a supplementary paper that has been deposited as a Supplementary Publication SUP 50158 (25 pages) at the British Library Document Supply Centre, Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem . J . (1990) 265, 5.

J Biol Chem, 1990 Sep 15, 265(26), 15867 - 73
Amino acid and cDNA sequence of bovine phosducin, a soluble phosphoprotein from photoreceptor cells; Lee RH et al.; Vertebrate photoreceptor cells contain a soluble phosphoprotein, phosducin, which complexes with the beta, gamma subunits of the GTP-binding protein, transducin . Light-induced changes in cyclic nucleotide levels modulate the phosphorylation of phosducin by protein kinase A . The complete amino acid sequence of purified phosducin from bovine retinas was determined by Edman degradation from overlapping polypeptides derived from enzymatic digestion by trypsin and Staphylococcus aureus V8 protease or from chemical degradation by cyanogen bromide . Excluding the unidentified group which blocks the NH2 terminus, phosducin contains 245 amino acids with a calculated molecular weight of 28,185 and isoelectric point of pH 4.5 . Phosducin is enriched with acidic and sulfur-containing amino acids, having 32 glutamic acid, 16 aspartic acid, 9 methionine, and 5 cysteine residues . It also contains 24 serine and 8 threonine residues, of which only serine 73 is located within a consensus phosphorylation sequence (-RKMS(P)QV-) for cyclic nucleotide-dependent protein kinase . Secondary structure analysis predicts the presence of 62% alpha-helix, 22% beta-sheet, and 16% random coil, with eight turns . Computer-aided searches of protein data banks revealed no apparent homology to any sequenced protein except that coded by a MEKA cDNA clone (Kuo, C-H., Akiyama, M., and Miki, N . (1989) Mol . Brain Res . 6, 1-10) which deviates from the confirmed phosducin sequence in the last 15 amino acids . Sequence analysis of a cDNA clone for bovine retinal phosducin confirmed that the MEKA clone deviation resulted from an unidentified cDNA guanosine nucleotide, a shifted reading frame and a premature stop codon.

J Immunol, 1990 Sep 15, 145(6), 1681 - 6
Frequency analysis of human peripheral blood B cells producing IgM-rheumatoid factor . Differential effects of stimulation with monoclonal antibodies to CD3 and Staphylococcus aureus; Hirohata S et al.; The extent and nature of IgM-rheumatoid factor (RF) precursors within normal human B cells were examined by utilizing two different polyclonal B cell stimulators, Staphylococcus aureus Cowan I (SA) and immobilized mAb to the CD3 molecular complex (64.1) . In cultures stimulated with SA, B cells produced IgM-RF in the presence of T4 cells, factors generated from mitogen-activated T cells (TF), or IL-2 . Similarly, in cultures stimulated with immobilized anti-CD3, T4 cells that had been treated with mitomycin C (T4 mito) induced the production of large amounts of IgM-RF . Limiting dilution analyses revealed that the precursor frequencies of IgM-RF-producing cells induced by SA + TF and by immobilized anti-CD3-activated T4 mito were 0.008 +/- 0.001/100 B cells (n = 7) and 0.043 +/- 0.004/100 B cells (n = 6) (mean +/- SEM), respectively . Of note, the proportion of IgM-secreting cells that produced IgM-RF was much greater in cultures stimulated with SA + TF (30 to 61%) than that noted in cultures containing immobilized anti-CD3-stimulated T4 mito (1.0 to 3.9%) . When B cells were co-stimulated with both SA and immobilized anti-CD3-activated T4 mito, the frequency of IgM-RF producing cells increased further to 0.12 to 0.27/100 B cells (4.6 to 21.2% of IgM-producing cells) . These results indicate that both SA and immobilized anti-CD3 are potent stimulators of IgM-RF precursors . Moreover, the combination of SA and immobilized anti-CD3 provides a very potent in vitro signal for IgM-RF elaboration, inducing the production of this autoantibody from 1 to 3 in 1000 circulating normal B cells.

Arch Surg, 1990 Sep, 125(9), 1177 - 80
The effect of venous obstruction in infected pedicle flap; Mann R et al.; A new model of soft-tissue infection is used to investigate the effect of the local wound environment on the septic focus . Island pedicle flaps were raised on the buttock of 24 adult ewes and multiply inoculated with Staphylococcus aureus . Flaps with bacterial inoculation, without compromise of venous outflow, showed distal necrosis (mean +/- SEM percent of surface area, 25.8% +/- 8.6%) and developed septic foci with bacterial counts one log less than the amount injected . Flaps with inoculation and venous outflow obstruction underwent subtotal necrosis (mean percent of surface area, 73.3% +/- 11.2%) and had counts two logs higher than the nonobstructed flaps but without discrete septic foci . Flaps without inoculation, with or without venous obstruction, survived completely . Venous outflow obstruction is shown herein to potentiate tissue necrosis by raising bacterial counts in a septic focus and preventing defensive abscess formation by the host.

Am J Kidney Dis, 1990 Sep, 16(3), 252 - 5
Percutaneous cyst puncture in the treatment of cyst infection in autosomal dominant polycystic kidney disease; Chapman AB et al.; Cyst infection in autosomal dominant polycystic kidney disease (ADPKD) poses a difficult diagnostic and therapeutic problem . We describe a serious indolent cyst infection due to Staphylococcus aureus, which was successfully diagnosed and treated with repeated percutaneous cyst drainage and intravenous (IV) antibiotic therapy . Cyst aspiration also permitted monitoring of cyst antibiotic levels and evidence of active infection.

Am J Kidney Dis, 1990 Sep, 16(3), 216 - 23
The role of continuous ambulatory peritoneal dialysis in end-stage renal failure due to multiple myeloma; Korzets A et al.; A study in 10 patients (eight male, two female; mean age 61.9 +/- 10.7 years) suffering from multiple myeloma (MM) and end-stage renal failure (ESRF) is detailed . Continuous ambulatory peritoneal dialysis (CAPD) was the preferred mode of chronic dialysis in all the patients . Survival after diagnosis was 32.2 +/- 23.9 months . Survival after starting dialysis was 24.6 +/- 20.6 months . All patients on CAPD were adequately dialyzed and in good fluid control . Peritonitis was the main problem on CAPD (one episode per 5.6 patient-months) . The majority of peritonitis episodes responded to intraperitoneal antibiotic therapy . One patient with Staphylococcus aureus peritonitis, septicemia, and neutropenia secondary to chemotherapy, died . Recommendations for prophylaxis and treatment of peritonitis are given . Three patients were transferred to hemodialysis . The use of subclavian vein catheters during hemodialysis was associated with a high incidence of gram-positive septicemia . Alkylating agent-based chemotherapy resulted in hematological responses in five patients . Survival after diagnosis in those responders was 47.4 +/- 25.6 months, compared with 17.0 +/- 7.2 months in the nonresponders (P less than 0.05) . All responders subsequently relapsed . Four patients died with progressive myeloma . Bone marrow suppression resulted in a high blood transfusion requirement, neutropenia, and thrombocytopenia associated with bleeding into the gastrointestinal tract and central nervous system . Uremic myeloma patients can be adequately dialyzed using CAPD . Those patients who do not have an initial hematological response have a poor prognosis.

J Bone Joint Surg Am, 1990 Sep, 72(8), 1240 - 4
Pyomyositis in a temperate climate . Presentation, diagnosis, and treatment; Hall RL et al.; The cases of eighteen patients who were treated for pyomyositis between 1970 and 1988 were evaluated . The diagnosis was often delayed because other primary diagnoses were considered, including muscle strain, synovitis, thrombophlebitis, and neoplasm, and because the symptoms were vague and prolonged (maximum duration, one year) . The muscles around the hip and thigh were most commonly involved (twelve patients), and Staphylococcus aureus most commonly grew on culture (twelve patients) . Computed tomography aided in the accurate diagnosis of the infection and of the extent of involvement . Incision, drainage, and antibiotic therapy eradicated the infection in all patients, and they had no residual functional limitations and minimum residual symptoms.

Am J Dis Child, 1990 Sep, 144(9), 1026 - 30
Bronchiolitis in tropical south India; Cherian T et al.; In a prospective hospital-based study of 328 children under 5 years of age with acute lower respiratory infections, 114 (35%) were diagnosed to have acute bronchiolitis . Of them, 87 (76%) were less than 1 year and 107 (94%) were less than 2 years of age . Signs of severe lower respiratory infections, namely tachypnea (respiratory rate greater than 50/min) and subcostal retraction, were present in 95% and 93%, respectively . Of 88 children of whom roentgenographs were taken, 30 (34%) had evidence of pneumonia . No clinical signs discriminated between those with and without pneumonia . By culture or immunofluorescence antigen detection, viruses were found in 81 (71%) children with bronchiolitis; respiratory syncytial virus was the most common agent, found in 65 (57%) . Parainfluenza viruses were the next most common, found in 12 (11%) . Most cases of bronchiolitis occurred in outbreaks during the rainy months of August through November, coinciding with respiratory syncytial virus outbreaks . Although bacterial culture of blood was done in 56 children, no respiratory pathogen was isolated . In one child with bronchiolitis and consolidation, postmortem lung aspirate yielded Staphylococcus aureus . Thus, bronchiolitis is primarily a viral syndrome in this tropical region, just as it is in temperate regions . Eight (7%) children died (all were infants); 5 had roentgenographic pneumonia and the remaining had other abnormalities contributing to death; all had been treated with antibiotics . Since one third of lower respiratory infections are bronchiolitis, and among infants under 1 year of age bronchiolitis comprises 47% of all lower respiratory infection cases, criteria for antibiotic management must take into account the availability of roentgenographic investigation.

Lab Invest, 1990 Sep, 63(3), 341 - 9
Staphylococcal alpha-toxin-induced vascular leakage in isolated perfused rabbit lungs; Seeger W et al.; alpha-Toxin, the major, pore-forming exotoxin of Staphylococcus aureus, caused acute hypertension when perfused through blood-free rabbit lungs (21) . This reaction is mediated by pulmonary thromboxane generation, for which toxin-induced calcium flux into target cells with subsequent stimulation of arachidonic acid metabolism is predominantly responsible . In the present study, we investigated the effects of alpha-toxin on the integrity of the lung microvasculature . Thromboxane generation was inhibited in all experiments to suppress the development of pulmonary hypertension . Application of low alpha-toxin concentrations (5 to 40 ng/ml) induced protracted, severe vascular leakage in a dose-dependent manner . After a lag period of 40 to 120 minutes, gravimetrically determined capillary filtration coefficients progressively increased to greater than 10-fold values, and this was followed by pronounced weight gain of the isolated organs . These physiologic alterations were paralleled by dose- and time-dependent structural changes documented by electron microscopic examination of perfusion-fixed lungs . Increasing electron density of microvascular endothelial cell nuclei and subsequently of their cytoplasma was noted, followed by detachment of these cells from the mutual endoepithelial basal lamina . Edema was localized in the blood-gas exchange area, in contrast to hydrostatically induced lung fluid accumulation . These results identify pulmonary microvascular endothelium cells as highly susceptible targets for attack by alpha-toxin . Given a similar sensitivity of human endothelial cells, alpha-toxin might directly contribute to the pathogenesis of acute respiratory failure under conditions of severe infection with Staphylococcus aureus.

J Clin Invest, 1990 Sep, 86(3), 942 - 51
Neutrophil bactericidal activity against Staphylococcus aureus adherent on biological surfaces . Surface-bound extracellular matrix proteins activate intracellular killing by oxygen-dependent and -independent mechanisms; Hermann M et al.; The activation patterns of surface adherent neutrophils are modulated via interaction of extracellular matrix proteins with neutrophil integrins . To evaluate neutrophil bactericidal activity, Staphylococcus aureus adherent to biological surfaces were incubated with neutrophils and serum, and the survival of surface bacteria was determined . When compared to albumin-coated surfaces, the bactericidal activity of neutrophils adherent to purified human extracellular matrix was markedly enhanced (mean survival: 34.2% +/- 9.0% of albumin, P less than 0.0001) despite similar efficient ingestion of extracellular bacteria . Enhancement of killing was observed when surfaces were coated with purified constituents of extracellular matrix, i.e., fibronectin, fibrinogen, laminin, vitronectin, or type IV collagen . In addition to matrix proteins, the tetrapeptide RGDS (the sequence recognized by integrins) crosslinked to surface bound albumin was also active (survival: 74.5% +/- 5.5% of albumin, P less than 0.02), and fibronectin-increased killing was inhibited by soluble RGDS . Chemiluminescence measurements and experiments with CGD neutrophils revealed that both oxygen-dependent and -independent bactericidal mechanisms are involved . In conclusion, matrix proteins enhance intracellular bactericidal activity of adherent neutrophils, presumably by integrin recognition of RGDS-containing ligands . These results indicate a role for extracellular matrix proteins in the enhancement of the host defense against pyogenic infections.

Crit Care Med, 1990 Sep, 18(9), 974 - 9
Effect of hyperbaric oxygen on murine neutrophil and T-lymphocyte functions; Gadd MA et al.; We investigated the effect of repeated hyperbaric oxygen (HBO) exposure on PMN and T-lymphocyte functions in a murine model . Animals received eight 90-min exposures twice daily to 2.4 ata and 100% or 10% oxygen . Control animals were maintained in room air . On the ninth day spleens and peritoneal cell exudates were harvested . Phagocytosis was measured by flow-cytometric analysis of the ability of PMN to engulf formalin-killed, fluorescence-labeled Staphylococcus aureus . PMN-killing capacity was measured by the ability of PMN to undergo an oxidative burst after stimulation with N-formyl methionyl-leucyl-phenylalanine, phorbol myristate, or opsonized zymosan . T-lymphocyte subpopulations were identified using monoclonal antibodies and two-color flow cytometry after 48 h stimulation with phytohemagglutinin, and lymphocyte proliferation was measured by 3 H-thymidine incorporation . We found PMN phagocytosis and oxidative burst were unchanged after HBO treatment . Lymphocyte proliferation was decreased, and an activated population of CD8+ T cells appeared after mitogen stimulation . We conclude that, although PMN function is not affected by prior HBO, lymphocyte proliferation is decreased.

Ann Emerg Med, 1990 Sep, 19(9), 987 - 90
The effect of local anesthetics on bacterial proliferation: TAC versus lidocaine; Martin JR et al.; We compared the effect of topical 0.5% tetracaine, 1:2,000 epinephrine, and 11.8% cocaine (TAC) with 1% lidocaine infiltration on bacterial proliferation in experimental lacerations . Forty-eight lacerations were made on the backs of Hampshire pigs, inoculated by injection with infectious doses of Staphylococcus aureus and randomly anesthetized with either topical TAC or lidocaine infiltration . Wounds were sutured, and quantitative cultures were obtained by excision after 48 hours . The mean log10 bacteria per gram of tissue for wounds anesthetized with TAC was 6.818 (95% confidence interval {CI}, 6.07 to 7.54) compared with 6.820 (95% CI, 5.91 to 7.75) for those treated with lidocaine; this difference was not significant (P less than .05 by paired two-tailed t test) . The probability of failing to detect an intergroup difference of 0.5 log10 bacteria per gram was less than .0001 . TAC does not increase bacterial proliferation more than lidocaine infiltration in contaminated experimental porcine lacerations.

J Infect Dis, 1990 Sep, 162(3), 759 - 62
Characterization of a widespread strain of methicillin-susceptible Staphylococcus aureus associated with nosocomial infections; McMurray LW et al.; Thirty Staphylococcus aureus isolates from five hospitals were determined to exhibit borderline susceptibility to the antistaphylococcal penicillins . Of the isolates submitted for phage typing, 96% belonged to phage group 94/96, and 96% possessed a common plasmid, pBW15 . Also, four reference borderline-susceptible isolates from the Centers for Disease Control belonged to phage group 94/96 and possessed pBW15 . Screening of 43 other phage group 94/96 isolates demonstrated that 36 (84%) contained pBW15 and exhibited the borderline phenotype . In contrast, pBW15 was not identified among 10 penicillin-susceptible, 10 methicillin-resistant, and 40 penicillin-resistant but non-borderline-susceptible S . aureus . These data show a close association between pBW15, phage group 94/96, and the borderline-susceptible phenotype . Furthermore, these isolates were associated with infections in multiple institutions, suggesting the widespread dissemination of a clinically important and pathogenic strain of S . aureus.

J Infect Dis, 1990 Sep, 162(3), 705 - 10
Binding affinity for penicillin-binding protein 2a correlates with in vivo activity of beta-lactam antibiotics against methicillin-resistant Staphylococcus aureus; Chambers HF et al.; The beta-lactam antibiotics ticarcillin, nafcillin, imipenem, and ampicillin, which differ in antibacterial activity against methicillin-resistant strains of Staphylococcus aureus, were examined for affinity to penicillin-binding protein (PBP) 2a, which mediates methicillin resistance . The relative efficacy of each antibiotic was compared to vancomycin in a rabbit model of aortic valve endocarditis caused by either a methicillin-susceptible or methicillin-resistant strain of beta-lactamase-producing S . aureus . beta-lactamase inhibitors clavulanate and sulbactam were used in combination with ticarcillin and ampicillin, respectively . All beta-lactam antibiotics were effective against the susceptible strain . beta-lactam antibiotic activity in vitro and in vivo against the resistant strain correlated with its affinity for binding to PBP 2a . Lack of efficacy of beta-lactam antibiotics for the resistant strain was due to an inability to eradicate the resistant subpopulation of cells . Vancomycin was the most effective agent.

Cell Immunol, 1990 Sep, 129(2), 341 - 50
Human T cell leukemia virus type I induces DNA synthesis and immunoglobulin secretion in human lymphocyte cultures; Boyer V et al.; Viral particles obtained from HTLV-I (human T cell leukemia virus, type I)-transformed T cell lines induced immunoglobulin production by normal peripheral blood lymphocytes . Conversely, no immunoglobulin could be detected in the supernatant medium in purified B cells cultivated with HTLV-I, suggesting that the presence of T cells is mandatory for HTLV-I to induce B cell polyclonal activation . The T cell help was mediated by soluble factors, as indicated in experiments showing that cell-free conditioned medium from T lymphocytes activated by HTLV-I was able to induce B cell proliferation and differentiation . Furthermore, a direct effect of HTLV-I on B cell proliferation was demonstrated when viral particles were added to purified B cells together with suboptimal doses of Staphylococcus aureus Cowan strain I (SAC) . These observations show that an immediate early effect of HTLV-I infection was exerted on B cells, mainly in a T cell-dependent manner . Such an effect may account for the hypergammaglobulinemia observed in HTLV-I seropositive individuals, and in patients with HTLV-I-associated neurological disorders.

Antimicrob Agents Chemother, 1990 Sep, 34(9), 1862 - 3
Increasing resistance of Staphylococcus aureus to ciprofloxacin; Daum TE et al.; We demonstrated the marked emergence of resistance to ciprofloxacin among Staphylococcus arueus strains isolated at the Ann Arbor Veterans Administration Medical Center . All S . aureus isolates tested from 1984 to 1985 were susceptible, whereas 55.1% of methicillin-resistant and 2.5% of methicillin-susceptible strains from 1989 had high-level resistance to ciprofloxacin.

Antimicrob Agents Chemother, 1990 Sep, 34(9), 1703 - 6
Evidence of a novel staphylococcal mec-encoded element (mecR) controlling expression of penicillin-binding protein 2'; Tesch W et al.; A region was identified on the methicillin resistance determinant (mec) isolated from Staphylococcus epidermidis and cloned into Staphylococcus carnosus which was responsible for a novel downregulation of the expression of methicillin resistance . The presence of this region reduced the overall expression of methicillin resistance and the synthesis of the mec-encoded penicillin-binding protein 2' (PBP 2') in S . carnosus . This region was located by Bal31 deletion mutagenesis upstream of the structural gene for PBP 2' . Deletions within this region resulted in higher levels of expression of methicillin resistance and increased levels of PBP 2' synthesis . We tentatively called this region mecR . Analysis of selected Mcr strains of Staphylococcus aureus and S . epidermidis by Southern hybridization suggested that the natural occurrence of two types of mec resistance determinants differ by the presence or absence of mecR-specific sequences.

Antimicrob Agents Chemother, 1990 Sep, 34(9), 1655 - 9
Efficacy of short courses of oral novobiocin-rifampin in eradicating carrier state of methicillin-resistant Staphylococcus aureus and in vitro killing studies of clinical isolates; Arathoon EG et al.; Methicillin-resistant Staphylococcus aureus (MRSA) is an important nosocomial infection problem . Colonization appears to be more common than invasive disease is . Eradication of colonization or the carrier state could limit the spread of MRSA, thus reducing the potential for mortality and morbidity in other patients . The detection of patients with MRSA infection in a rehabilitation ward led to a study of the combination of novobiocin-rifampin in vivo and in vitro . We found that 300 mg of rifampin plus 500 mg of novobiocin orally twice daily for 5 days, in 18 courses of treatment given to 12 patients, resulted in the clearing of MRSA in 79% of the evaluable courses and 81% of the evaluable sites . A second course cleared MRSA from one of the patients with a treatment failure . Side effects were not noted . All 18 pretherapy isolates were susceptible to either drug in vitro, but 1 of 2 posttherapy isolates was rifampin resistant . Timed-kill studies demonstrated that the rate of killing was the same with either drug alone or both drugs together . Pretherapy isolates from treatment successes or failure were killed at the same rate by the drug combination . However, with the rifampin-resistant isolate killing ceased after 48 h . Results of this study suggest that previously untreated patients are likely to have isolates that are susceptible to the combination of drugs and that the combination is commonly effective in eradicating MRSA carriage . Since the regimen is orally administered, and thus convenient, in conjunction with other measures it has the promise of reducing the spread of MRSA in hospitals.

FEMS Microbiol Lett, 1990 Sep 1, 59(1-2), 55 - 63
Characterization of a small cryptic plasmid isolated from a methicillin-resistant strain of Staphylococcus aureus; Walters JA et al.; The nucleotide sequence of a small (1613 bp) plasmid, pOX2000, isolated from a methicillin-resistant strain of Staphylococcus aureus has been determined . The sequence contains only one large ORF and the predicted amino acid sequence shows homology to the REP proteins of some other small staphylococcal plasmids . In addition there are two palindromic sequences, palA and palJ, that are similar to but not identical with the palindromes known from other staphylococcal plasmids to be involved in lagging strand initiation and possibly leading strand termination, respectively . Preliminary functional analysis of pOX2000 has been carried out by assessing the effect of interrupting the sequence at three unique restriction endonuclease sites . The plasmid pOX2000, and its relationship to other small staphylococcal plasmids, is discussed.

Indian J Med Res, 1990 Sep, 91, 349 - 54
Monocyte/macrophage functions & humoral response in blood & bronchoalveolar lavage fluid of pulmonary tuberculosis patients; Goyal M et al.; Subsegmental bronchoalveolar lavage (BAL) was performed in 33 patients with active pulmonary tuberculosis and five control subjects . Phagocytosis by monocytes and alveolar macrophages was studied, and in addition serum and BAL immunoglobulin and complement levels were also determined . The phagocytic activity of blood monocytes was depressed in pulmonary tuberculosis patients as compared to controls, 37.8 +/- 2.3 per cent; 50.7 +/- 4.2 per cent and 32.9 +/- 3.6 per cent for sheep RBC's, latex and Staphylococcus aureus respectively compared to 66.7 +/- 6, 54.8 +/- 2.2 and 68.3 +/- 3.5 per cent respectively in controls; the differences being significant for sheep RBC's (P less than 0.05) and Staph . aureus (P less than 0.001) . However, phagocytosis was not impaired in BAL macrophages (P greater than 0.05) . In patients no significant alteration in serum immunoglobulin and complement levels was observed except that levels of C4 component of complement were increased in patients with far advanced lesions (98.5 +/- 33.7 mg/dl compared to 78.7 +/- 7.9 mg/dl; P less than 0.05) . While IgM and C4 component of complement could not be detected in BAL fluid the levels of IgA were significantly increased in pulmonary tuberculosis patients (65.5 +/- 50.5 mg/dl compared to 39.9 +/- 13.3 mg/dl in control; P less than 0.05) . Since IgA secreted in the BAL fluid is mostly synthesised locally, increased levels of this immunoglobulin could be of value in determining activity of the disease.

Nippon Saikingaku Zasshi, 1990 Sep, 45(5), 845 - 9
{Induction by Staphylococcus aureus L-form of tumor necrosis factor-alpha from macrophages}; Matsu-Ura I et al.; Induction of tumor necrosis factor-alpha (TNF-alpha) by Staphylococcus aureus L-form was investigated . The supernatant of a macrophage culture mixed with S . aureus L-form showed a potent cytotoxic activity to L cells . Addition of anti TNF-alpha antibody inhibited completely the cytotoxic activity of the supernatant, indicating that the activity might be due mostly to TNF-alpha . To investigate localization of TNF-alpha production, the membranes of hypotonicity treated L-form were layered on a step-gradient composed of an upper and lower layers of 35% and 50% sucrose, respectively . The membranes were banded at the interface of 35% and 50% of sucrose . The activity of TNF-alpha production of the membrane fraction was 10-times higher than that of the soluble fraction.

J Virol Methods, 1990 Sep, 29(3), 279 - 89
An indirect sandwich ELISA utilising F(ab')2 fragments for the detection of African horsesickness virus; du Plessis DH et al.; African horsesickness virus (AHSV), an important disease of equines is caused by an orbivirus . Because of the need to contain the spread of the disease, it is often essential to make a rapid diagnosis . For this purpose, an ELISA capable of detecting viral antigen in animal tissue and in cell culture fluid was developed . Immobilised F(ab')2 fragments prepared by digestion of AHSV-specific IgG with pepsin were used to trap virus from tissue homogenates or cell culture supernatant . After addition of intact IgG as detecting antibody, Staphylococcus aureus protein A labelled with horseradish peroxidase was added to allow visualisation of the reaction . Polyclonal antibodies directed against either whole AHSV or viral core particles were suitable as detecting antibodies . On the other hand, a monoclonal antibody that was specific for a major core protein, VP7, gave a much weaker signal in the ELISA . All known AHSV serotypes were recognised in the F(ab')2-ELISA by polyclonal antisera against either whole virus particles or viral cores . Immunoprecipitation of AHSV structural polypeptides showed that such antisera contained populations of antibodies directed against core proteins . The F(ab')2-ELISA has potential as a diagnostic technique for AHSV infections.

Jpn J Antibiot, 1990 Sep, 43(9), 1621 - 8
{A clinical study on S6472, sustained release preparation of cefaclor, dermatological area}; Kawanami T et al.; We conducted a clinical investigation on the use of S6472, an sustained release preparation of cefaclor, in the dermatological area . Thirty-two patients serving as subjects were divided into group I (14 cases, including folliculitis and acne pustulora), group II (16 cases, including furuncle, furunculosis and carbuncle) and others (2 cases) . One or 2 wrappers (375 or 750 mg) of S6472 were orally administered to patients twice daily after meals . In group I, the efficacy rate was 100% . Clinical results were as follows: Excellent (6 cases), good (8 cases), fair (0), and poor (0) . In group II, the efficacy rate was 93.8%: Excellent (10 cases), good (5 cases), fair (1 case), and poor (0) . A decrease of the bacteriological effect was observed in 1 case with Staphylococcus epidermidis . A superinfection was noted in 1 case Staphylococcus aureus . The causative bacteria were all eradicated in the remaining 24 cases . No side effects were detected . From the above results, S6472 appears to be a useful preparation, twice daily oral administration has excellent effects against dermatological infections.

Yakugaku Zasshi, 1990 Sep, 110(9), 658 - 64
{Studies on FK482 (Cefdinir) . III . Synthesis and structure-activity relationships of 7 beta-{(Z)-2-aryl-2-hydroxyiminoacetamido}-3-vinyl-3- cephem-4-carboxylic acid derivatives}; Inamoto Y et al.; The synthesis, antibacterial activity and oral absorption of the 7 beta-{(Z)-2-aryl-2-hydroxyiminoacetamido}-3-vinylcephalosporins (Ia--e) are described . All of these compounds exhibited excellent activity against Staphylococcus aureus . Against Gram-negative bacteria FK482 exhibited more excellent activity than the other compounds (Ia--e) . These compounds except Ie showed good oral absorption . The relationship between the oral absorption rates and the lipophilicity of these cephalosporins is discussed.

J Dairy Sci, 1990 Sep, 73(9), 2372 - 8
Relationships among vitamin E, selenium, and bovine blood neutrophils; Hogan JS et al.; Effects of vitamin E and selenium supplementation on in vitro phagocytosis and intracellular kill of bacteria by bovine neutrophils were investigated . Diets were not supplemented with vitamin E and selenium during the dry period and first 21 d of lactation . Cows were then assigned to one of four treatment diets for 30 d . Treatment diets were either unsupplemented or supplemented with vitamin E, selenium, or both vitamin E and selenium, in a 2 x 2 factorial arrangement . Peripheral blood neutrophils were isolated from each cow on lactation d 51 . Vitamin E supplementation of diets increased intracellular kill of Staphylococcus aureus and Escherichia coli by neutrophils . Intracellular kill of S . aureus was greater in neutrophils isolated from selenium supplemented cows than in neutrophils from cows without supplemental selenium . Intracellular kill of E . coli did not differ between neutrophils from selenium supplemented and selenium unsupplemented cows . Ability of neutrophils to phagocytize either S . aureus or E . coli was independent of vitamin E and selenium.

FEMS Microbiol Immunol, 1990 Sep, 2(2), 65 - 73
Detection of Staphylococcus aureus infection by enzyme-linked immunosorbent assay and immunoblotting, using high molecular weight staphylococcal proteins; Ryden C et al.; Two high molecular weight staphylococcal proteins, fibronectin-binding protein and a Mr 200,000 protein, were investigated as antigens for serodiagnosis of staphylococcal infections . Sera from patients with staphylococcal infections and from controls were subjected to immunoblot analysis with staphylococcal lysate proteins to identify staphylococcal antigens to which patients with staphylococcal infections specifically exhibited antibodies . One such protein was found in the Mr 200,000 region . This protein was purified and used as antigen in ELISA and compared with other antigens, namely fibronectin-binding protein(s) (FNBP, Mr 185,000), alpha-toxin and teichoic acid . Sera from patients with staphylococcal infections contained antibodies to the high molecular weight proteins in higher titers than sera from patients with non-staphylococcal infections or healthy subjects . Based on their amino-acid compositions and different abilities to bind fibronectin it was concluded that the Mr 200,000 protein and FNBP were not identical.

Zh Mikrobiol Epidemiol Immunobiol, 1990 Sep, (9), 80 - 5
{The characteristics of the protective and antigenic activities of purified staphylococcal anatoxin in trials with mice of different genotypes}; Semenova IB et al.; The protective potency of purified staphylococcal toxoid by the survival rate of immunized mice challenged with the culture of Staphylococcus aureus strain L-1726 and the antigenic properties of the toxoid, determined by the level of antitoxin in the blood of mice and by the intensity of cell-mediated immunity in the spleen-cell migration inhibition test, were studied . The experiments were made on CBA and C57BL/6 mice . Purified staphylococcal toxoid was shown to possess antigenic and protective properties in a wide range of doses between 0.15 and 15 binding units per mouse . The protective effect of the toxoid in CBA mice was manifested in the presence of circulating antibodies and cell-mediated reaction or only in the presence of the toxoid . In C57BL/6 mice the protective effect of the toxoid was less pronounced and appeared in combination with the induction of cell-mediated immunity in the presence of an extremely low antibody level (0.062 I.U.).

Semin Respir Infect, 1990 Sep, 5(3), 226 - 32
Approach to therapy of respiratory infections in the critical care setting; Pennington JE; Therapy of pneumonia in the critical care setting includes intravenous antibiotics and supportive care . Since the etiologic agent of infection may not be clear, empiric broad-spectrum antibiotic regimens are often used . Combinations of beta-lactam and aminoglycoside agents are particularly popular regimens due to the high incidence of gram-negative bacillary and Staphylococcus aureus pneumonias in the critical care unit . Several new approaches to treatment of pneumonia in the critical care setting are being evaluated, including single-agent empiric coverage using a broad-spectrum beta-lactam agent; broad-spectrum quinolones, such as ciprofloxacin; intrabronchial aminoglycoside instillation therapy; and passive immune therapy with immunoglobulins and monoclonal antibodies.

Semin Respir Infect, 1990 Sep, 5(3), 157 - 72
Nosocomial pneumonia in the 1990s: update of epidemiology and risk factors; Craven DE et al.; Hospital-acquired pneumonia is the second most common nosocomial infection in the United States . Aspiration appears to be the major route for the entry of microorganisms into the lower respiratory tract . Nosocomial pneumonia may be caused by bacteria, viruses and fungi . Aerobic gram-negative bacilli and Staphylococcus aureus are the most common etiologic agents, but infection is usually polymicrobial . Risk factors for nosocomial pneumonia include host variables, colonization with nosocomial pathogens, and impaired response of pulmonary defenses to the microbial challenge . Bacteria causing nosocomial pneumonia may be part of the patient's endogenous flora, originate from the hands of hospital personnel, or result from the use of invasive devices . The mechanically ventilated patient has multiple risk factors that contribute to the high rate of nosocomial pneumonia . An understanding of the epidemiology and risk factors for nosocomial pneumonia is fundamental for implementation of preventive strategies to reduce patient morbidity, mortality, and hospital costs.

Vet Immunol Immunopathol, 1990 Sep, 26(1), 1 - 12
Selective alteration of bovine neutrophil responses by recombinant bovine interleukin-1 beta; Canning PC et al.; The effects of recombinant bovine interleukin-1 beta (rBIL-1 beta) upon in vitro bovine neutrophil functions were determined . Exposure of peripheral blood neutrophils to various concentrations of rBIL-1 beta induced dose dependent suppression of the phagocyte's ability to migrate under agarose . Preincubation of neutrophils with rBIL-1 beta did not influence their ability to ingest radiolabelled Staphylococcus aureus nor did it induce hydrogen peroxide production or elastase release . However, pretreatment of phagocytes with rBIL-1 beta did result in a dose-dependent enhancement of opsonized zymosan-induced H2O2 production . In contrast, rBIL-1 beta had no effect upon the ability of opsonized zymosan-stimulated neutrophils to release elastase from primary granules . Pretreatment of neutrophils with rBIL-1 beta for as little as 15 min was sufficient to induce suppression of migration and enhancement of opsonized zymosan-induced H2O2 production . These results suggest rBIL-1 beta is capable of directly modulating selected neutrophil activities . In addition, rBIL-1 beta appears to augment the phagocyte's oxidative metabolic responses to subsequent stimulation by microbial antigens.

J Microsc, 1990 Sep, 159 ( Pt 3), 277 - 83
Flow cytometric analysis of the functional ability of neutrophils from patients with autoimmune neutropenia; Macey MG et al.; Flow cytometry has been used to evaluate the functional ability of neutrophils and the expression of IgG Fc receptors (FcRII and FcRIII) in autoimmune neutropenia . Quantification of the neutrophil oxidative burst was made by assaying the production of 2'7'-dichlorofluorescein (DCF) from non-fluorescent 2'7'-dichlorofluorescein trapped within the cell, by flow cytometric analysis of cellular fluorescence . In the present study the DCF assay was used to examine the response of neutrophils to stimulation by opsonized and non-opsonized Staphylococcus aureus . In addition, the rate of uptake of S . aureus labelled with the red nuclear dye propidium iodide was determined . The presence of surface-bound immunoglobulin, which may affect the phagocytic capacity of the neutrophil, was also measured . No correlation between the neutrophil count and level of membrane-bound IgG or the rate of bacterial uptake was found . The studies were performed on twenty patients with autoimmune neutropenia, twelve with other autoimmune disorders and fourteen normal controls . The rate of uptake of bacteria was considered in relation to the expression of FcRII and FcRIII . Good correlation was found with the level of expression of FcRII, the major receptor for neutrophil activation, and the rate of uptake of bacteria (r = 0.64).

Am J Otol, 1990 Sep, 11(5), 347 - 53
Hyperbaric oxygen management of chronic staphylococcal osteomyelitis of the temporal bone; Schweitzer VG; Hyperbaric oxygen (HBO) has proven efficacious in the adjunct management of selected otolaryngologic problems: radiation therapy-induced osteoradionecrosis of the temporal bone, malignant external otitis, mandibular osteoradionecrosis and refractory osteomyelitis, soft tissue head and neck necrotizing fasciitis, compromised skin flaps and grafts, acute air or gas embolism, and otologic barotrauma . Herein is described the management of an insidious Staphylococcus aureus osteomyelitis of the temporal bone by pre- and postoperative HBO in conjunction with surgical debridement . The possible application of angiogenic agents and tetracycline bone-labeling in combination with hyperbaric oxygen therapy in the management of refractory neurotologic disease will be discussed.

Kokyu To Junkan, 1990 Sep, 38(9), 903 - 7
{A case of acute pericarditis secondary to mediastinitis}; Kawai S et al.; A 45-year-old man who complained of swallowing disturbance and chest pain in inspiration phase was admitted for evaluation of "pericarditis" . A chest X-ray film on admission disclosed a wide mediastinal shadow and pleural effusion on the right side . Bilateral tonsils were swollen, and covered with pus . A Computed tomogram of the chest showed a shadow of exudate contained with air in the mediastinum . Mediastinal drainage, tonsillectomy and drainage of fistula from pre-tracheal space to upper mediastinum were immediately performed . Staphylococcus aureus was confirmed from the sputum and mediastinal effusion . Thus, acute mediastinitis was confirmed as an etiological diagnosis of "pericarditis" in this patient.

Rinsho Byori, 1990 Sep, 38(9), 998 - 1004
{Methicillin-resistant Staphylococcus aureus infection in the Kagoshima University Hospital--special attention to prevalence in otolaryngological infectious disease}; Uchizono A et al.; A retrospective study of the characteristics of MRSA (methicillin resistant S . aureus) in Kagoshima University Hospital and its ENT department was reported . There were 231 samples (104 strains) MRSA in all S . aureus 448 samples from Jan . to Sep . 1989 in Kagoshima University Hospital, in which 58 strains (55.8%) were coagulase type II and 35 strains (33.7%) were type VII . The much more predominant numbers of MRSA were observed in all the cases inpatients clinics in surgery, pediatrics, urology, gynecology and dermatology than internal medicine as well as outpatients clinics . In ENT department of this hospital, only 15.4% of all S . aureus were MRSA . On the other hand, bacteriological survey showed 26% of MRSA out of all S . aureus which were detected at Tokyo clinical research center on otorrhea in the patients with chronic otitis media as multicenter clinical trial of Japan . Two cases involved by MRSA after postoperative administration of many types of antibiotics were also demonstrated . These MRSAs were producing type II coagulase and very high concentrations of MICs to many antibiotics were observed . But these cases were not severe and MINO (minocyclin) or GM (gentamicin) were so effective . The effect of combined therapy of some antibiotics or antimicrobial agents was discussed briefly.

Rinsho Byori, 1990 Sep, 38(9), 990 - 7
{Nosocomial infection with methicillin-resistant Staphylococcus aureus}; Kusano N et al.; Methicillin-resistant Staphylococcus aureus (MRSA) isolated in our hospital between 1986 and 1989 were mainly examined for their susceptibility to various antibiotics and coagulase types . The isolation frequency of MRSA among S . aureus isolated from clinical specimens has been steadily increasing; 37.8% in 1986, 49.8% in 1987, 60.6% in 1988 and 63.2% in 1989 . This trend was particularly noticeable in the specimens associated with the respiratory tracts . The isolation rate of MRSA in the surgery and pediatrics wards was higher than that in the internal medicine ward . More than 80% of MRSA were coagulase type II, which were multi-resistant to penicillins, cephems, aminoglycosides and macrolides, and sensitive to MINO, new quinolones and VCM . These epidemic strains were also isolated from the nose of medical staff and from air samples in the wards . These findings suggest that the hospital environment including the patients and hospital personnel is extensively contaminated with multi-resistant MRSA of coagulase type II . Measures should be taken for prevention and control of nosocomial infection with MRSA in the whole hospital.

Rinsho Byori, 1990 Sep, 38(9), 983 - 9
{Mechanism of beta-lactam-resistance in methicillin-resistant Staphylococcus aureus}; Okonogi K; The role of penicillin-binding protein (PBP) 2' in the expression of beta-lactam-resistance was investigated using methicillin-resistant Staphylococcus aureus (MRSA) strains with different level of resistance . Both high- and moderate-level MRSA produced very similar PBP 2' with low affinities for beta-lactam antibiotics . Affinities of antibiotics for PBP 2' (I50, concentration which inhibits {14C} benzylpenicillin-binding by 50%) correlated well with their antibacterial activities (MIC) in a high-level MRSA, but did not in a moderate-level MRSA . High-level MRSA contained a larger amount of PBP 2' than moderate-level MRSA, and the amount of PBP 2' decreased by increasing the temperature of the culture; the extent of decrease was larger in a strain which was sensitive at 37 degrees C than a strain which exerted relatively high level resistance even at 40 degrees C . A cephamycin-resistant, methicillin-sensitive strain began to synthesize PBP 2' by adding cephamycin-type antibiotics to the medium and consequently acquired resistance to methicillin . Latent MRSA producing no PBP 2' generated clones which produced PBP 2' constitutively and were highly resistant to all beta-lactams . These results suggest that the presence of PBP 2' is critical for the expression of beta-lactam-resistance in MRSA and the degree of the resistance depends mainly on the amount of PBP 2' which differs from strain to strain and is influenced by environments such as temperature and the presence of inducer.

Rinsho Byori, 1990 Sep, 38(9), 1005 - 15
{Methicillin-resistant Staphylococcus aureus (MRSA) infection--significance of MRSA in respiratory tract infection}; Shigeno Y et al.; We have examined background factors in MRSA infection in cases in which S . aureus had been isolated from sputa . The incidence of isolation of S . aureus was high and still increasing in expectorated sputa, and causative organisms in the cases of pneumonia and autopsied lungs . A significant correlation was observed between high incidence of isolation of S . aureus and abuse of third-generation cephems . MRSA isolation rates of inpatients was higher than that of outpatients . Among the inpatients such cases with severe underlying diseases and prolonged admission showed the highest incidence of isolation of MRSA . There seemed to be a correlation between distribution of patients with S . aureus and that of rooms with S . aureus in the air . This suggests nosocomial infection . Although MRSA was frequently isolated from sputa, most cases showed no signs of infection, and this suggested that they had been transient colonization . Such antimicrobial agents as rifampicin, teicoplanin, vancomycin reveal excellent antibacterial activity against MRSA and minocycline, ofloxacin were moderately effective . The physician must be informed of the significance of MRSA, because their understanding of MRSA still remains insufficient.

Infect Control Hosp Epidemiol, 1990 Sep, 11(9), 479 - 85
Frequent acquisition of multiple strains of methicillin-resistant Staphylococcus aureus by healthcare workers in an endemic hospital environment; Opal SM et al.; Methicillin-resistant Staphylococcus aureus (MRSA) has been an endemic nosocomial pathogen at the VA medical center (VAMC) in Providence, Rhode Island since 1981 . From 1985 to 1987, more than 30% of all unique S aureus isolates were methicillin resistant . To evaluate the frequency of acquisition of MRSA isolates by healthcare workers, we compared the antimicrobial susceptibility patterns, multilocus enzyme genotypes and plasmid profiles of isolates recovered from nasal and hand cultures from VAMC nurses and house staff on rotation at the VAMC with those of clinical isolates from patients at the VAMC and four other affiliated hospitals . Fifty-six percent of ward nurses cultured (n = 112) were colonized with S aureus, of which 65% was methicillin resistant . Six isolates of MRSA were identified on the initial culturing of house staff (n = 65); 16 MRSA isolates were recovered at the end of a four-week rotation (p less than .02) . Phenotypic and genotypic analyses demonstrated that numerous distinct MRSA strains were recovered in the study period . The incidence of MRSA among clinical isolates at the VAMC and affiliated institutions was remarkably constant throughout the three-year study period . Moreover, despite regularly sharing resident physicians, interns and medical students, MRSA isolates were commonly recovered at the other university-affiliated hospitals . Our study failed to reveal evidence of significant interhospital transmission of MRSA isolates by healthcare workers . While healthcare workers may contribute to the dissemination of MRSA within institutions, they appear to be less important in spreading MRSA between institutions.

Eur J Clin Microbiol Infect Dis, 1990 Sep, 9(9), 649 - 53
Use of a ribosomal RNA gene probe for the epidemiological study of methicillin and ciprofloxacin resistant Staphylococcus aureus; Hadorn K et al.; Conventional bacteriophage typing was combined with ribotyping in the analysis of methicillin and ciprofloxacin resistant Staphylococcus aureus strains isolated in increasing frequency since the introduction of the new 4-quinolones as therapeutic agents in the Tel-Aviv Medical Center . Whole-cell DNA was digested with EcoRI and HindIII restriction endonucleases . Agarose gel electrophoresis, Southern blotting, and hybridization by biotinylated probe DNA coding for ribosomal RNA revealed 7 to 14 bands . Analysis of the patterns established a single DNA type in EcoRI as well as in HindIII digests for all strains except one . Control strains from other sources differed in their band patterns . Bacteriophage typing confirmed the results of DNA typing . Thus, the frequent occurrence of staphylococcal isolates resistant to 4-quinolones in the hospital was not due to mutational development of resistance in many strains, but to the spread of a resistant strain.

Br Vet J, 1990 Sep-Oct, 146(5), 443 - 8
Characteristics of Staphylococcus aureus from subclinical bovine mastitis in Brazil; Lopes CA et al.; A total of 127 Staphylococcus aureus strains isolated from milk samples of cows with subclinical bovine mastitis was examined for biotype, phage pattern, in-vitro antibiotic susceptibilities and ability to produce enterotoxins . The majority of the strains showed features consistent with bovine rather than human origin . All strains were sensitive to the antibiotics tested, except penicillin and streptomycin . Enterotoxigenicity was observed in 6 (4.7%) strains and only enterotoxins A and C were produced.

Zentralbl Veterinarmed B, 1990 Sep, 37(7), 556 - 60
Milk whey induction of agglutination in ovine and bovine mastitis Staphylococcus aureus; Baselga R et al.; A total of 59 mastitis staphylococcic strains were tested for growth agglutination upon supplementation of growth media with ovine and bovine milk whey and mammary secretions from dry cows . Differences were observed when comparing bacterial species or origins (ovine vs . bovine) of bacteria and whey . All of the ovine and bovine S . aureus strains tested, but only 4 among 22 other ovine mastitis staphylococcic strains, showed growth agglutination in Todd Hewitt broth (THB) supplemented with greater than or equal to 30% (v/v) ovine milk whey . None of the strains agglutinated during growth in regular THB medium . Ovine whey had an agglutination induction capacity higher than bovine whey (P less than 0.005), concerning the number of responsive ovine and bovine S . aureus strains . There were no differences between whey samples from different ewes with regard to their capacity to induce agglutination . Ovine S . aureus strains were more responsive than bovine strains of this bacterial species, concerning the number of responsive strains (P less than 0.001) to bovine whey (greater than or equal to 30% in THB), the proportion of responsive strains at low (10%) ovine whey concentration (P less than 0.001), and the strength of reaction (precipitation timing and clump size) . Secretions from dry cows systematically induced agglutination in all of the bovine and ovine S . aureus strains tested.

South Med J, 1990 Sep, 83(9), 1092 - 5
Pyomyositis and human immunodeficiency virus infection; Blumberg HM et al.; Pyomyositis is a bacterial infection of skeletal muscle usually caused by Staphylococcus aureus and characterized by localized muscle pain, swelling, and tenderness . The disease is endemic in the tropics . Though only approximately 50 cases have been reported from the continental United States, pyomyositis has been increasingly recognized here in the last decade . We report two patients with human immunodeficiency virus (HIV) infection and pyomyositis, and review five previously reported cases . Given the predisposition of patients with the acquired immunodeficiency syndrome (AIDS) or AIDS-related complex (ARC) for infections caused by S aureus, pyomyositis may become increasingly more common in temperate areas.

Antimicrob Agents Chemother, 1990 Sep, 34(9), 1777 - 9
Insertional inactivation of the mec gene in a transposon mutant of a methicillin-resistant clinical isolate of Staphylococcus aureus; Matthews P et al.; All clinical strains of methicillin-resistant Staphylococcus aureus (MRSA) examined so far contain the mec gene and its product, the penicillin-binding protein (PBP) 2A . Yet the same strains show tremendous variation in the phenotypic expression of antibiotic resistance (MIC), which is under the control of a set of additional, auxiliary genes . Thus, the quantitative contribution of the mec gene to the resistance phenotype of MRSA is not known, and no mutants with the lesion located within the mec gene have been described . We subjected a highly resistant MRSA strain to transposon mutagenesis with the erythromycin resistance transposon Tn551, and a mutant expressing greatly decreased methicillin resistance (RUSA4) was selected to characterize the transposon insertion site . The results indicate that the Tn551 insertion site in mutant RUSA4 is between base pairs 1000 and 1400 of the sequence encoding PBP 2A . Thus, the uniform and greater than 200-fold drop in the methicillin MIC (4 micrograms/ml) for this mutant relative to that for the parent strain (MIC greater than or equal to 800 micrograms/ml) must be related to the inactivation of the PBP 2A gene . The results provide the first unequivocal evidence for the importance of PBP 2A as a quantitative contributor to the MIC for MRSA.

J Clin Microbiol, 1990 Sep, 28(9), 1898 - 902
Staphylococcus aureus colonization and infection in patients on continuous ambulatory peritoneal dialysis; Pignatari A et al.; Staphylococcus aureus is the most common cause of peritonitis in patients undergoing peritoneal dialysis in Brazil . Using restriction endonuclease analysis of plasmid DNA, we investigated the importance of chronic carriage of S . aureus in the development of peritonitis in patients on continuous ambulatory peritoneal dialysis at the Division of Nephrology, Escola Paulista de Medicina, Sao Paulo, Brazil . A total of 117 isolates (30 patients) of S . aureus were available for typing, including 51 isolates (22 patients) from the nares, 58 isolates (27 patients) from pericatheter skin, and 8 isolates (6 patients) from peritoneal fluid, from patients with peritonitis . Restriction endonuclease subtyping showed that although most patients harbored more than one subtype of S . aureus, in the majority of patients nasal and/or pericatheter skin isolates with identical restriction endonuclease digest patterns were recovered on more than one occasion . Furthermore, 95% of patients with both nasal and pericatheter colonization were colonized with the same subtypes at both sites . All of the patients with peritonitis were infected with a subtype which colonized the nares, pericatheter skin, or both . These results demonstrate the importance of an endogenous source of S . aureus in the development of continuous ambulatory peritoneal dialysis-associated peritonitis.

Prikl Biokhim Mikrobiol, 1990 Sep-Oct, 26(5), 602 - 8
{Isolation of SAU 96 I restrictase, its physical and catalytic properties}; Lebedev LR et al.; Restrictase Sau 96 I was isolated from Staphylococcus aureus PS 96 and purified by chromatography on DEAE-cellulose, phosphocellulose and hydroxylapatite . The preparation was studied by gel filtration on Toyopearl HW-55 and polyacrylamide gel electrophoresis under denaturing conditions . The active form of the enzyme is a dimer with a molecular weight of 54,000 +/- 5000 composed of two identical subunits . Catalytic properties of the restrictase were determine; the pH optimum is 8.5-9.0, the optimal concentration of NaCl and Mg2+ is 15-100 mM and 10 mM, respectively . Mn2+ ions at a concentration of 2 mM can replace Mg2+, while Zn2+, Ca2+, Cu2+ ions cannot replace Mg2+ . The optimal temperature is 30-43 degrees . Ethanol and glycerol at concentrations more than 10% inhibit the enzyme without changing its specificity; p-chloromercuribenzoate inhibits the enzyme at a concentration of 0.05 mM.

Zentralbl Bakteriol, 1990 Sep, 273(4), 518 - 30
Comparative study on the macrolides erythromycin and clarithromycin: antibacterial activity and influence on immune responses; Roszkowski K et al.; The in vitro activity of erythromycin and clarithromycin (a new macrolide antibiotic) on clinical bacterial isolates as well as their effects on the cellular and humoral immune responses in BALB/c-mice and on human granulocytes/monocytes was investigated . Treatment of BALB/c-mice for 7 days with these drugs did not influence the delayed type hypersensitivity to oxazolone nor the production of IgG and IgM immunoglobulins . In vitro, exposure of granulocytes to erythromycin resulted in increased phagocytosis only in higher concentrations (20 mcg/ml), whereas clarithromycin enhanced chemiluminescence response of granulocytes in concentrations ranging from 2.5-20 mcg/ml . Subinhibitory concentrations of both substances modified Staphylococcus aureus and made them more susceptible for granulocyte phagocytosis.

Infect Immun, 1990 Sep, 58(9), 2809 - 14
Pulmonary antibacterial defenses during mild and severe influenza virus infection; Nickerson CL et al.; Severe influenza virus infections with pneumonic involvement are known to predispose the lungs to bacterial superinfections due to dysfunctions in the alveolar macrophage (AM) phagocytic system . To determine whether milder forms of influenza without pneumonic involvement have a similar outcome, pulmonary antibacterial defenses and AM phagocytosis were compared in murine models of mild and severe influenza virus A/HK/68 infections . Bactericidal activity was quantitated by the intrapulmonary killing of Staphylococcus aureus following aerosol challenge, whereas the functional capacity of the AMs was determined by Fc-receptor-mediated phagocytosis . With the severe virus infection, maximal suppression of bactericidal activity occurred on day 8 of infection and correlated with impairment of AM phagocytosis . A lesser but significant degree of suppression of pulmonary antibacterial defenses and AM phagocytosis was observed on the third day of the mild virus infection . The data demonstrate that mild influenza virus infections that are limited to the upper respiratory tract also impair pulmonary antibacterial defenses and may predispose the lungs to bacterial superinfections.

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