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 Glu2