Microbiologica, 1992 Jan, 15(1), 75 - 8 Association of coagulase and/or pigmentation with the virulence of a capsule-lacking Staphylococcus aureus in iron-compromised mice; Salamah AA; The effect of pigmentation and coagulase on the virulence of a capsule-lacking Staphylococcus aureus were studied using iron-compromised mice . The presence of either coagulase or pigmentation maintained the viability of this bacterium in the peritoneal cavity . Both coagulase and pigmentation, however, lead not only to maintained viability, but also to its proliferation in the peritoneal macrophages . The iron was found to enhance the virulence of this bacterium, but iron enhancement was contingent on the presence of either pigmentation or coagulase or preferably both. Arerugi, 1992 Jan, 41(1), 65 - 72 {Autoimmune response to thyroglobulin . Proliferative response to thyroglobulin fragments in low responder mice}; Hoshioka A et al.; Thyroglobulin (Tg) is one of the major thyroid autoantigens involved in autoimmune thyroiditis . The immune response of mice to Tg is genetically controlled by H-2-linked genes . To elucidate the regulation mechanism of autoimmune response to Tg in low responder mice, we studied the proliferative response of lymph node cells (LNC) to mouse Tg (MTg) and enzyme-digested MTg fragments . MTg was treated with Staphylococcus aureus V8 protease followed by separation of the fragments into 6 fractions (Fr1-Fr6: 264,000-17,000) by high performance liquid chromatography (HPLC), LNC from MTg immunized CBA/N (H-2k) mice, a high responder strain, proliferated in response to MTg and all fractions (Fr1-Fr6) of MTg fragments in vitro . In contrast, LNC from MTg immunized BALB/c (H-2d) and B10 (H-2b) mice, low responder strains, did not respond to native Tg but responded well to some smaller Tg fractions (Fr3, 4, 5) . In addition, when BALB/c mice were immunized with MTg Fr4 with a molecular weight of 63,000, LNC from BALB/c mice proliferated in response to MTg as well as MTg Fr4 . These findings suggest that T cells which are capable of responding to Tg do exist even in low responder mice and that the activation of these autoreactive T cells is suppressed by a regulatory cell subpopulation in low responder mice. J Pediatr Surg, 1992 Jan, 27(1), 101 - 2 Anterior mediastinal abscess complicating septic arthritis; Smith A et al.; An 11-year-old boy with septic arthritis of both knees presented with an anterior mediastinal abscess extending suprasternally . This was drained through a suprasternal incision and the mediastinal cavity was intermittently irrigated with povidone iodine solution and packed with gauze . Staphylococcus aureus was the responsible organism . Antibiotic therapy comprised of cloxacillin and gentamycin . Recovery was uneventful . This is, most probably, the first report on an anterior mediastinal abscess complicating a distant septic arthritis . As for any infective mediastinitis, early diagnosis and aggressive treatment is mandatory for a patient's survival. J Clin Immunol, 1992 Jan, 12(1), 36 - 44 Interleukin 4 inhibits polyclonal immunoglobulin secretion and cytokine production by peripheral blood mononuclear cells from rheumatoid arthritis patients; Briolay J et al.; Rheumatoid arthritis (RA) is associated with T- and B-cell dysfunction . Immunoglobulin (Ig) production is under the control of T cells and their derived cytokines such as interleukin 2 (IL-2) and IL-4 . Herein we studied the regulation of the production of immunoglobulins and cytokines by peripheral blood mononuclear cells from RA patients and controls . In the controls, IL-4 inhibited Ig production in response to Staphylococcus aureus and pokeweed mitogen stimulation . IL-2 induced maximal Ig production in association with Staphylococcus aureus, whereas it inhibited pokeweed mitogen-induced production . In patients, levels of Ig production in response to mitogens and cytokines were reduced, particularly for the response to IL 2 . The inhibitory effect of IL-4 on mitogen-induced Ig production was observed in RA patients as in the controls . Spontaneous production of IL-6 was increased in RA patients . These levels were correlated with indicators of active disease such as sedimentation rate and Ritchie index . IL-4 inhibited the production of IL-6, IL-1 beta, and tumor necrosis factor alpha (TNF alpha) by both controls and rheumatoid patients . Thus as first described for the T-cell response, mononuclear cells from RA patients display a reduced response to mitogens and cytokines which induce their B-cell differentiation into Ig-screening cells . However, IL-4 was able to inhibit Ig and cytokine production, properties suggesting a potential antiinflammatory role for this cytokine. Arch Fr Pediatr, 1992 Jan, 49(1), 51 - 3 {Neonatal cervical cord compression by a staphylococcal abscess}; Desportes V et al.; In a small for date premature newborn infant, septicemia caused by Staphylococcus aureus was observed on day 16 . In spite of an appropriate antimicrobial therapy, proteins of inflammation serum levels remained elevated for 2 weeks . The occurrence of cervical cord compression which could be related to a staphylococcal abscess explained the persisting inflammatory process . After puncturing of the abscess, immobilization and prolonged antibiotic treatment, outcome was favorable . This case demonstrates the difficulty of preventing abscesses related to staphylococcal septicemias and reports a very unusual site of abscess which could be well localized using MRI. Med Microbiol Immunol (Berl), 1992, 180(6), 273 - 8 Recombinant epidermolytic (exfoliative) toxin A of Staphylococcus aureus is not a superantigen; Fleischer B et al.; The epidermolytic (exfoliative) toxins produced by Staphylococcus aureus cause epidermolysis and skin blistering . In addition, they have been implicated to belong to the group of T lymphocyte stimulating molecules known as "superantigens" . Here we show that recombinant epidermolytic toxin A produced in S . aureus is not mitogenic for human and murine T lymphocytes . We discuss the possibility that minute contaminations of highly mitogenic exoproteins may cause the mitogenicity in several proteins that are reported to be superantigens. Medicine (Baltimore), 1992 Jan, 71(1), 52 - 7 Duration of fever during treatment of infective endocarditis; Lederman MM et al.; We reviewed the duration of fever among 123 patients treated for infective endocarditis at University Hospitals of Cleveland between 1972 and 1984 . One half of these patients became afebrile within 3 days after initiation of antibiotic therapy and nearly three quarters were afebrile after 1 week of therapy . After 2 weeks of therapy, nearly 90% had defervesced . Endocarditis due to Staphylococcus aureus or gram-negative bacilli, and culture-negative endocarditis, were associated with prolonged fever . Microvascular phenomena, major vessel embolization, or vegetations seen on 2-D echocardiogram also were associated with prolonged fever . Multivariate analysis revealed that only microvascular phenomena or major vessel embolization were independently associated with longer duration of fever . Endocarditis-associated mortality among patients who remained febrile after 1 week of therapy was 18%, and this was greater than the 2% mortality among patients who defervesced (p less than 0.002) . These data suggest that prolonged fever during treatment of infective endocarditis is often due to tissue infarction or vascular injury . Prolonged fever also identifies patients at higher risk of a fatal outcome. FEMS Microbiol Immunol, 1992 Jan, 4(2), 105 - 10 Antibody response to Staphylococcus aureus whole cell, lipase and staphylolysin in patients with S . aureus infections; Ryding U et al.; Three assays to measure antibodies against Staphylococcus aureus whole cells, lipase and staphylolysin were used to try to discriminate between complicated and uncomplicated S . aureus septicaemia . Sera were examined from 8 patients with S . aureus endocarditis, 23 patients with complicated S . aureus septicaemia, 12 patients with uncomplicated S . aureus septicaemia and 93 febrile non-septicaemic controls . No single assay could distinguish between complicated and uncomplicated S . aureus septicaemia . If the criterion for a positive result is defined as positive antibody level in the anti-lipase ELISA as well as in at least 1 of the other 2 assays, 10/31 patients with S . aureus endocarditis or complicated septicaemia were positive compared to 0/93 non-septicaemic patients and 0/12 patients with uncomplicated S . aureus septicaemia . Therefore, the combined use of serological assays in the diagnosis of complicated S . aureus septicaemia, one of which is the anti-lipase ELISA, is recommended. Infect Control Hosp Epidemiol, 1992 Jan, 13(1), 33 - 7 Clonal diversity of methicillin-resistant Staphylococcus aureus in an acute-care institution; Nicolle LE et al.; OBJECTIVE: To evaluate plasmid and chromosomal typing methods for differentiation of methicillin-resistant Staphylococcus aureus (MRSA) . DESIGN: Comparison of relatedness of strains using epidemiologic features, phage typing, and antimicrobial susceptibility with blinded assessment by molecular typing methods . Molecular typing methods included AccI and ClaI restriction endonuclease fingerprinting of chromosomal DNA and plasmid typing . SETTING: Tertiary-care teaching hospital . METHODS: Convenience sample of 10 MRSA strains, including 4 outbreak-associated and 6 sporadic strains of diverse epidemiologic origins without evidence of nosocomial transmission . RESULTS: Only 2 strains were phage typeable . The epidemic strain was distinguishable by its susceptibility pattern . The other strains were not consistently separable by phenotyping or plasmid typing methods but were distinct by chromosome typing . CONCLUSIONS: These observations document the diversity of origins of MRSA strains introduced into an acute-care institution . Chromosomal typing may be the preferred method for the determination of clonal origin of MRSA. J Dairy Sci, 1992 Jan, 75(1), 66 - 71 Colonization by Staphylococcus aureus on chapped teat skin: effect of iodine and chlorhexidine postmilking disinfectants; Fox LK; The abilities of common postmilking teat disinfectant solutions and a teat skin ointment to retard Staphylococcus aureus colonization and promote healing of chapped skin were studied . Ten Holstein cows, free of S . aureus intramammary infections and mammary epidermal colonization, were used . All teats were experimentally chapped with 1N sodium hydroxide and dipped in S . aureus broth culture of 5 x 10(6) cfu/ml after three consecutive milkings . Postmilking teat disinfectant treatments were applied, starting after the second application of S . aureus broth culture, and were continued for the next 19 milkings . Quarters of each cow received one of the following postmilking teat treatments: 1% iodine plus 10% glycerin solution; .5% chlorhexidine plus 4.87% glycerin solution; 1% chlorhexidine ointment; and no postmilking treatment . After milking and before treatment application, samples of skin swabbings of the teat end and teat side and samples of milk were collected . Daily teat skin chapping visual scores were made to assess chapping and healing . Teat skin chapping decreased with time, and the decline was similar among treatments . The concentration of S . aureus recovered from teat skin swabbing solutions decreased over time . Staphylococcus aureus were recovered at significantly lower concentrations from swabbing solutions of teats that were dipped with iodine and chlorhexidine solutions as compared with untreated teats . Staphylococcus aureus was in marginally higher concentrations on ointment-treated teats than on dipped teats . Results indicate the importance of using postmilking teat dips on chapped teats and suggest no advantage in applying a disinfectant ointment to chapped teats to reduce S . aureus colonization and improve skin healing. J Dairy Sci, 1992 Jan, 75(1), 19 - 26 Growth of Staphylococcus aureus and synthesis of enterotoxin during ripening of experimental Manchego-type cheese; Gomez-Lucia E et al.; To study the possible presence of staphylococcal enterotoxins in Manchego-type cheese, milk was inoculated with the enterotoxigenic Staphylococcus aureus collection strains FRI-100, S6, FRI-137, and FRI-472 to a final concentration of 10,000 to 25,000 cfu/ml . Cheese was prepared following the industrial specifications and ripened for 60 d . Batches were prepared with 1 and .1% lactic acid culture and labeled with the abbreviated name of the strain and the concentration of lactic acid culture . Mean staphylococcal counts in .1% lactic bacteria cheeses were usually more than 1 log higher than the corresponding 1% ones . Staphylococcal counts declined markedly after d 35 to 42, and, by the end of ripening, they had disappeared from some cheeses . Enterotoxins were present in five of the cheeses, three prepared with .1% and two with 1% lactic bacteria . Enterotoxins detected were A and D, the enterotoxins most commonly associated with human intoxication . The maximum level of enterotoxin A detected in cheese with strain FRI-100 and with the .1% culture was 222 ng/100 g of cheese; in cheese FRI-100 with 1%, 111 ng/100 g; in cheese S6 with .1%, 769 ng/100 g; and in cheese S6 with 1%, 33 ng/100 g . Maximum level of enterotoxin D detected in cheese FRI-472 with .1% was 38 ng/100 g. J Appl Bacteriol, 1992 Jan, 72(1), 16 - 20 Enterotoxin production in different Staphylococcus aureus biotypes isolated from food and meat plants; Isigidi BK et al.; Strains of Staphylococcus aureus isolated in Belgium and Zaire from food and from various sources in the meat industry were biotyped, phage typed and tested for staphylococcal enterotoxin (SE) production . Thirty of the 185 strains examined produced one or more SE, and 23 of these belonged to the human biotype . Most SE-positive strains belonged to phage groups III and Mixed, or were not typable . None of the poultry-like biotype strains, which were frequent in nasal carriers among workers in meat plants as well as in minced meat, produced enterotoxins . Avian biotype strains similarly were negative. Appl Environ Microbiol, 1992 Jan, 58(1), 6 - 13 Staphylococcal coagglutination, a rapid method of identifying infectious hematopoietic necrosis virus; Bootland LM et al.; A staphylococcal coagglutination test was developed for the rapid detection of infectious hematopoietic necrosis virus (IHNV) in cell cultures and infected fish . The test could be completed in 15 min but required a minimum IHNV titer of 10(6) PFU/ml to obtain a positive reaction . All IHNV isolates, representing the five electropherotypes taken from a wide variety of species and different geographic ranges, caused coagglutination of Staphylococcus aureus cells sensitized with rabbit polyclonal serum against the Round Butte IHNV isolate . The coagglutination reaction was blocked by preincubation of IHNV with homologous antiserum, and IHNV did not cause coagglutination of S . aureus cells sensitized with normal rabbit serum . In specificity tests, cells sensitized with rabbit anti-IHNV serum or normal serum did not coagglutinate in the presence of infectious pancreatic necrosis virus, viral hemorrhagic septicemia virus, cell culture medium components, or media from cultures of cell lines of salmonid and nonsalmonid origin . Most importantly, the coagglutination test was able to detect and identify IHNV directly from experimentally infected rainbow trout fry, the organs of naturally infected adult kokanee salmon and winter steelhead trout, and ovarian fluids of the winter steelhead trout . The coagglutination test is very suitable for field use, since it is inexpensive, simple to interpret, sensitive, and rapid and requires no specialized equipment. Appl Environ Microbiol, 1992 Jan, 58(1), 403 - 4 Novel assay for screening rifamycin B-producing mutants; Vohra RM; A simple method that allows easy identification of rifamycin B-producing strains is described . This method involves the use of an enzyme, rifamycin oxidase, which converts inactive rifamycin B to active rifamycin S . In this method, colonies to be tested are grown in pairs . The two colonies are then transferred to two plates seeded with a sensitive strain of Staphylococcus aureus, one plate of which contains the enzyme rifamycin oxidase . All paired colonies which show a larger inhibition zone diameter on the enzyme-containing plate are identified as rifamycin B producers. Ann Pediatr (Paris), 1992 Jan, 39(1), 27 - 36 {Surveillance of antibiotic therapy in a pediatric intensive care unit}; Borderon JC et al.; Since 1982, a pediatric intensive care unit for neonates and pediatric patients up to 15 years of age has prospectively recorded every instance of use of antimicrobials, with the reasons for use, clinical and bacteriological parameters, and outcome . This approach encourages strict adherence to established protocols and provides a basis for discussing the rationale of each antimicrobial course . Effectiveness of protocols is evaluated annually and modifications or additions are introduced, as appropriate . Patterns in the proportion and nature of antimicrobials used to treat hospital-acquired infections can be monitored on the basis of the data collected . Changes in the nature of antimicrobials used, which may have repercussions on pathogen resistance to antimicrobials, are also monitored . Reasons for use of antimicrobials are categorized as follows: A = primary infection: B = secondary infection acquired in the ICU; C: secondary infection acquired in another unit or in another hospital; D = prophylaxis . In 1987, 46% of neonates and 59% of patients above one month of age were given antimicrobial agents; these figures are similar to those recorded during the previous years . Reasons for antimicrobial therapy were as follows in neonates: A = 48.5%; B = 40%; C = 1.2%; D = 10.3%; in patients above one month of age corresponding figures were: A = 23%; B = 44%; C = 0.9%; D = 31.9% . Among the neonates, the ampicillin-aminoglycoside combinations accounted for 41.5% of treatments (1/4th of these treatments were continued); in the older patients, penicillin G and ampicillin were the most commonly used antibiotics . In all age groups, hospital-acquired infections were mainly respiratory tract infections (approximately 50% in neonates and 80% in patients above one month of age) . Staphylococcus aureus was the most prevalent organism; Pseudomonas was seen virtually only among the patients above one month of age with very prolonged endotracheal intubation . From 1983 through 1987, use of third-generation cephalosporins increased from 4.5% to 28.3% in neonates and from 5.5% to 9% for patients above one month of age . The changes identified over time should be interpreted in the light of changing patterns of disease; in particular, hospital-acquired infections among neonates increased twofold, probably as a result of the rising number of very-low-birth-weight infants. FEMS Microbiol Lett, 1992 Jan 1, 69(2), 161 - 4 An efficient microtest to study adherence of bacteria to mammalian cells; Aguilar B et al.; A simple, fast and highly reproducible microtest was developed for in vitro adherence studies . A rat epithelial cell line was investigated for the adherence of clinical and subclinical ovine and bovine Staphylococcus aureus strains isolated from mastitis . Staphylococcus aureus strains differed in their ability to adhere to epithelial cells, the degree of adherence being dependent on the concentration of bacteria used in the test. J Cardiovasc Surg (Torino), 1992 Jan-Feb, 33(1), 70 - 4 Prevention of prosthetic vascular graft infection by rifampicin impregnation of a protein-sealed Dacron graft in combination with parenteral cephalosporin; Avramovic J et al.; Antibiotic-impregnated prosthetic vascular grafts have been shown to be highly resistant to bacterial contamination in animal models . The aim of this study was to assess if graft infection in an animal model challenged with local bacterial contamination could be reduced further by the use of rifampicin soaking of a protein-sealed Dacron graft in addition to the prophylaxis provided by perioperative intravenous cephalosporin . Of 20 Merino sheep given cefoxitin intravenously before the induction of anaesthesia, ten had a rifampicin treated Dacron graft implanted into the common carotid artery (group 1), and ten received an untreated Dacron graft (group 2) . Before wound closure the grafts were inoculated with 1 ml of 10(8) colony forming units per ml of Staphylococcus aureus . After three weeks the grafts were removed and cultured . Group 1 sheep had fewer graft infections (two of 10) compared to group 2 (six of eight survivors), this difference being statistically significant (p = 0.03; Fisher exact test) . It is concluded that rifampicin treatment of protein-sealed Dacron grafts combined with intravenous cefoxitin provides more protection from contaminating Staphylococcus aureus than intravenous cefoxitin alone in this animal model. Int Immunol, 1992 Jan, 4(1), 43 - 51 Distinct antigen presenting cell-derived signals induce TH cell proliferation and expression of effector cytokines; Schmitz J et al.; In addition to the stimulus delivered by the specific interaction of the T cell receptor (TCR) and the antigen - MHC class II complex, activation of resting helper T lymphocytes (TH) requires several poorly defined accessory cell-derived co-stimulatory signals . Here we provide evidence that proliferation and expression of effector cytokine genes by TH cells are induced by distinct co-stimulatory signals . Normal murine splenic TH cells were activated by Staphylococcus aureus enterotoxin B (SEB) superantigen and various antigen presenting cells (APCs) to proliferate, differentiate into TH cells blasts, and secrete cytokines . Blast transformation and proliferation of TH cells is achieved with macrophages and other splenic APCs, like B cells . Expression of the cytokines interferon gamma (IFN gamma), IL-5, and IL-2 by TH cells, however, is to various degrees dependent on the presentation of SEB by macrophages . The requirement for macrophages is particularly striking for the expression of IFN gamma . Thus macrophages provide distinct co-stimulatory signals for cytokine secretion and proliferation . The results suggest that B cells induce clonal expansion of TH cells whereas macrophages additionally promote terminal differentiation of activated TH cells into TH effector cells. J Vasc Surg, 1992 Jan, 15(1), 187 - 93 Prevention of graft infection by bonding of gentamycin to Dacron prostheses; Haverich A et al.; To increase the efficacy of perioperative antibiotic prophylaxis in vascular surgery an experimental study including topical application of the gentamicin derivative EMD 46/217 and fibrin sealant as antibiotic carrier to Dacron prostheses was initiated . In vitro treatment of Dacron with gentamicin and fibrin was followed by constant antibiotic release for 3 weeks . In a subsequent animal study Dacron grafts were implanted in the aorta of 10 pigs after direct contamination with Staphylococcus aureus solution . One graft was pretreated with the antibiotic/fibrin compound, a second with the antibiotic alone . Grafts 3 (no pretreatment) and 4 (fibrin alone) served as controls . After 1 week the grafts and their corresponding implantation sites were excised for measurement of antibiotic content and for culture . The antibiotic content of grafts with the antibiotic/fibrin compound was 25.0 +/- 7.2 micrograms/gm wet weight, whereas Dacron pretreated with the antibiotic alone contained no measurable drug amounts except for one specimen (0.5 microgram/gm) (antibiotic/fibrin vs antibiotic, p less than .0005) . The corresponding implantation sites to antibiotic/fibrin grafts contained 1.07 +/- 0.54 microgram/gm antibiotic, whereas in only 2/10 implantation sites of antibiotic grafts low antibiotic levels were found (0.05 and 0.2 microgram/gm) (antibiotic/fibrin vs antibiotic, p less than 0.005) . All control grafts and 9/10 antibiotic grafts were infected . By contrast, only five were contaminated, and 5 of 10 remained sterile after culture (antibiotic/fibrin vs antibiotic, p less than 0.05) . This finding correlates with the antibiotic content in the Dacron . It is concluded that pretreatment of prosthetic Dacron grafts with the antibiotic/fibrin compound results in binding of sufficient amounts of antibiotic for at least 1 week.(ABSTRACT TRUNCATED AT 250 WORDS) J Immunol, 1992 Jan 1, 148(1), 1 - 6 Localization of the immunologic activity in the superantigen Staphylococcal enterotoxin B using truncated recombinant fusion proteins; Buelow R et al.; The exotoxins of certain strains of Staphylococcus aureus strains are able both to stimulate potent proliferation and induce anergy in T lymphocytes expressing the appropriate T cell Ag receptor V beta gene elements . Although T cell activation by the S . aureus enterotoxins requires the presence of accessory cells bearing class II Ag of the MHC, unlike the peptide fragments of nominal Ag, they contact the external surfaces of both the class II MHC and TCR molecules . This paper investigates the immunologically active domains of S . aureus enterotoxin B (SEB) using truncated fragments of rSEB expressed as a fusion protein with protein A . The results of the experiments reported here indicate that the minimal fragment of SEB able to stimulate and induce anergy in hemagglutinin-reactive human T cells expressing V beta 3.1 gene elements is located in the amino-terminal portion of the molecule within residues 1-138 . Deletion of the first 30 amino acid residues renders rSEB unable to stimulate T cells expressing V beta 3.1, whereas polyclonal T cells still respond to this molecule . This implies that the stimulation of several TCR-V beta families may be caused by the interaction with different regions of the toxin . The localization of immunologically active sites in the bacterial enterotoxins is needed to investigate both their biology and potential application as immunomodulatory agents. Adv Exp Med Biol, 1992, 311, 389 - 90 Effects of low ionic strength and calmodulin on the decline in maximal calcium-activated force in permeabilized smooth muscle; Crichton CA et al.; Maximal calcium-activated tension (Tmax) was measured from saponin-and alpha-toxin permeabilized strips of rat anococcygeus muscle stimulated repetitively . 50 mins after saponin treatment, Tmax had declined to 20% of the initial value . Calmodulin (1 microM) only partially reduced this decline . Lowering the ionic strength of the bathing solution from 0.2M to 0.07M markedly reduced the rate of decline of tension . Addition of calmodulin to the bathing medium of low ionic strength prevented the decline of force still further; 50 mins after permeabilization, Tmax declined to 80% of the original level . Loss of endogenous calmodulin should be prevented by permeabilization with alpha-toxin from Staphylococcus aureus . Nevertheless, Tmax still declined to approximately 50% of the original value after 50mins . However, negligible force reduction was observed if the bathing solution had an ionic strength of 0.07M. Microbiol Immunol, 1992, 36(5), 545 - 50 Conspicuous ingestion of Staphylococcus aureus organisms by murine fibroblasts in vitro; Usui A et al.; A conspicuous adhesion of Staphylococcus aureus organisms to murine cutaneous fibroblasts and NIH/3T3 cells cultured in vitro and subsequent ingestion of S . aureus organisms by these fibroblasts are described . In the present experimental system, only fibroblasts-adhering S . aureus organisms were efficiently ingested by fibroblasts unlike S . epidermidis and S . saprophyticus . These findings might suggest a correlation between the pathogenesis of S . aureus and its intracellular localization in non-professional phagocytes such as fibroblasts in a special reference to its higher pathogenicity than those of coagulase negative counterparts. Microbiol Immunol, 1992, 36(5), 445 - 53 Analysis of borderline-resistant strains of methicillin-resistant Staphylococcus aureus using polymerase chain reaction; Hiramatsu K et al.; Identification of methicillin-resistant Staphylococcus aureus by drug-susceptibility tests alone poses a serious problem, because a considerable number of clinical S . aureus isolates are borderline resistant to methicillin . To circumvent this problem, we have developed a quick and sensitive method of PCR amplification for the detection of mecA gene, which, coding for PBP2', is the specific genetic element of methicillin-resistant Staphylococcus aureus . This method made it possible to identify MRSA strains in a short time using as few as 30 cells as a starting material for template DNA . Using this method, we found that the strains of borderline methicillin-resistance could be accurately identified . We also found one S . aureus clinical strain, T3, which lacked mecA gene in spite of its resistance to methicillin. Microbiol Immunol, 1992, 36(5), 431 - 43 Intracellular localization of Staphylococcus aureus within primary cultured mouse kidney cells; Murai M et al.; Staphylococcus aureus Cowan I was incubated with monolayers of cells derived from several portions of mouse kidney, and found to be ingested by all types of the renal cells . Intracellular localization of S . aureus was determined by resistance of intracellular cocci against lysostaphin digestion and confirmed by electron microscopy . From renal medulla, three morphological variants of the hyperosmolarity-tolerant (HOT) cells were obtained . The rate of cocci-ingesting cells varied from 16.9% to 93.4% among these of the HOT cells at the end of 3-hr incubation . From renal cortex, three morphological variants of epithelial cells grew in medium RK-1 . Among them, only the cells on the edge of colony ingested Cowan I, while the epithelial cells on the center of colony ingested few cocci . Transferred from medium RK-1 to MEM supplemented with 10% FBS, part of the cortical cells changed into fibroblast-like appearance and obtained the capacity to ingest Cowan I . This result may indicate the correlation between ingesting capacity and cellular morphology . From a glomerulus, epithelial (GE) cells and fibroblast-like (GF) cells were obtained . The GE cells ingested not only S . aureus Cowan I but Staphylococcus epidermidis and Staphylococcus saprophyticus after 30-min incubation, while the GF cells, like both of the HOT cells and the cortical cells, ingested only S . aureus . These results suggest a possibility that S . aureus is located within nonprofessional phagocytes during its infection and intracellular coccus plays an important role in its pathogenicity. Chemotherapy, 1992, 38(3), 179 - 84 In vivo postantibiotic effect of isepamicin and other aminoglycosides in a thigh infection model in neutropenic mice; Minguez Minguez F et al.; The object of this work is to study in neutropenic mice the in vivo postantibiotic effect (PAE) of isepamicin, a new aminoglycoside, gentamicin and netilmicin on Staphylococcus aureus and Escherichia coli and in vivo killing kinetics using two different schedules (A and B) of isepamicin and gentamicin administration against S . aureus: (A) at time zero and every hour up to the 7th or 9th hour and (B) two doses only, at time zero and at the end of the PAE . The PAE of the three aminoglycosides was long (3-5 h), showing that of isepamicin to be the largest, especially on S . aureus . Both A and B treatment models show the same effectiveness for the two tested drugs . These results support the idea that the major significance of the PAE is in its application to dosing regimens. Arch Microbiol, 1992, 157(3), 242 - 8 Lethal and mutational effects of solar and UV radiation on Staphylococcus aureus; Chapple RM et al.; Strains of Staphylococcus aureus, an opportunistic pathogen commonly found on human skin, were exposed to sunlight and UV C radiation, and the lethal and mutational effects measured . Sunlight killed cells with an inactivation constant of 3 x 10(-5) per joule per square metre; UV C was much more lethal, giving an inactivation constant of approximately 0.1 per joule per square metre . Some strains tested showed a sensitivity to sunlight that was dependent on the growth phase of the cells, exponentially growing cells showing a greater sensitivity . Mutational effects of irradiation were measured by the appearance of mutants sensitive to methicillin following irradiation of a multiresistant strain . Mutants appeared at a frequency of 10(-3); this high frequency of mutation in the region of the mec gene has also been observed when multiresistant strains are subjected to nutritional or thermal stress . Mutants showed the same chromosomal alteration (seen in pulse-field gel electrophoresis of SmaI-digested DNA) whether induced by solar or UV C irradiation. J Orthop Trauma, 1992, 6(4), 401 - 6 The use of antibiotic-impregnated polymethylmethacrylate beads to prevent the evolution of localized infection; Seligson D et al.; The effect of antibiotic bead chains on the evolution of infection cannot be studied entirely in man due to the ethical problems of obtaining valid controls . Therefore, a model of musculoskeletal injury was devised in rabbits by making a paraspinal wound, fracturing a spinous process, and contaminating the wound with 1 x 10(4) colony-forming units/ml of Staphylococcus aureus . These contaminated wounds were treated with tobramycin-containing polymethylmethacrylate (PMMA) beads . Control rabbits were either treated with PMMA beads that did not contain antibiotic, treated with IM tobramycin, or left untreated . At 5 days, six of eight animals treated with antibiotic-impregnated beads had no recoverable organisms . Six of eight rabbits receiving IM tobramycin had wound infections, and five of five in whom non-antibiotic-containing beads had been implanted had significant wound infections, with one of the five dying of sepsis on the 3rd day of the experiment . The clinical course of infected controls was the same as the course of those animals receiving IM antibiotics and the same as those in whom beads without antibiotics were implanted . That is, the rabbits had grossly infected wounds and the organisms recovered were of the same type as those implanted . This research shows a highly statistically significant effect of tobramycin-containing antibiotic beads in retarding the evolution of an experimental Staphylococcus infection in rabbits. Respiration, 1992, 59(5), 265 - 71 Enhanced chemiluminescence with decreased antibody-dependent cellular cytotoxicity of human alveolar neutrophil in patients with adult respiratory distress syndrome; Lin CC et al.; In order to elucidate the intrinsic defects of inflammatory cells in adult respiratory distress syndrome (ARDS), 10 ARDS patients, 6 non-ARDS respiratory failure patients and 30 healthy controls were included in this study . The peripheral blood neutrophils (PBNs) were isolated from heparinized venous blood . Human alveolar neutrophils (HANs) were taken during bronchoalveolar lavage (BAL) . Chemiluminescence was then measured with a luminometer (LKB Wallac) and antibody dependent cellular cytotoxicity (ADCC) was assayed using a Cr51 cytotoxic assay . The bacterial killing test utilized heparinized PBNs and Staphylococcus aureus . The results showed that the chemiluminescence of PBNs and HANs was higher in ARDS patients than in the normal control group (p < 0.05) . However, ADCC levels in the PBNs and HANs of ARDS patients were much lower than in normal subjects (p < 0.01) . The bacterial killing ability of PBNs was also impaired in ARDS patients, compared with either the normal control subjects or non-ARDS respiratory failure patients . In conclusion, chemiluminescence is increased in both PBNs and HANs of ARDS patients, which could partly explain the pathogenesis of acute lung injury . Furthermore, the suppression of ADCC in both PBNs and HANs as well as the impaired bacterial killing of PBNs may explain the impaired bacterial defenses and the consequent increase in pulmonary infection among ARDS patients. Zh Mikrobiol Epidemiol Immunobiol, 1992, (9-10), 63 - 5 {The effect of the structural components of Staphylococcus aureus on the interaction of T- and B-lymphocytes}; Pozur VK et al.; The results of studies on the influence of staphylococcal peptidoglycan and cell walls on the cooperative interaction of T- and B-lymphocytes in the process of immune response to thymus-dependent antigen are presented . Peptidoglycan has been found to produce, depending on its dose, a suppressive and stimulating effect on the interaction of T- and B-cells . Cell walls exhibit only stimulating action under the same experimental conditions . The suppressive action of peptidoglycan is mediated by T-lymphocytes. Acta Derm Venereol Suppl (Stockh), 1992, 176, 132 - 4 Comparative effects of two topical antiseptics (chlorhexidine vs KMn04) on bacterial skin flora in atopic dermatitis; Stalder JF et al.; In order to determine the efficacy and tolerance of two topical antiseptics, chlorhexidine vs KMn04 (diluted at 1:20,000), we compared their bacteriological and clinical effects in a randomized trial on 20 children with Atopic Dermatitis (AD) treated with topical steroids (desonide) . After treatment, a clinical improvement was noted in the two groups, though without statistical differences . In vivo: Before treatment, Staphylococcus aureus (S.A.) density was high and predominant in both groups . After treatment, the decrease in S.A . was greater in the chlorhexidine group than in the KMn04 group, without significant difference . In vitro: At the clinical dilution used, there was a statistical difference (p < 0.05) between the number of killed bacteria in the chlorhexidine group (-3 log) and the number in the KMn04 group (-1 log) . This study confirms the role and importance of the choice of a topical antiseptic in the treatment of AD. Int Orthop, 1992, 16(4), 330 - 8 {One stage revision of infected total hip replacements with replacement of bone loss by allografts . Study of 90 cases of which 46 used bone allografts}; Loty B et al.; We usually manage infected total hip replacements by a one stage revision . We have reviewed 90 cases operated on before 1988 . Notable bone loss was frequently encountered and reconstructed using allografts . In all cases complete surgical debridement was performed and a cemented total hip replacement implanted . Bone allografts were used in 46 cases . Systemic antibiotics and antibiotic loaded cement were utilised . There were 17 failures (19%); one patient died soon after operation, there were 8 obvious infections (9%), 1 hip was thought to be infected although the prosthesis was not loose, and 7 femoral prostheses became loose of which 3 were due to mechanical failure and the remainder had no obvious explanation . Infection with staphylococcus aureus and pseudomonas had a statistical correlation with failure, but other factors including the preoperative status and the use of allografts did not appear to influence the result . Allograft femoral heads from the bone bank were found to be reliable for reconstruction of the acetabulum and small femoral defects . With major femoral bone loss we preferred massive irradiated cortical allografts . The quality of the bone reconstruction was probably the most important factor in the good functional results observed in 79% of cases . Effective surgical debridement and an appropriate antibiotic regime will allow a successful one stage revision procedure . The use of allografts gave a reliable reconstruction of the bone loss and was not associated with an increased rate of failure. Chemotherapy, 1992, 38(4), 211 - 7 Influence of exogenous factors and antibiotics on the cytoplasmic membrane proteins of Staphylococcus aureus; Cullmann W et al.; The influence of various nutrient media, growth temperature, phase of growth and subinhibitory concentrations of antibiotics from various groups on the cytoplasmic membrane proteins was studied in 6 clinical S . aureus isolates; further study included the comparison of the cytoplasmic membrane proteins in antibiotic-sensitive wild-type (parent) strains with their resistant counterparts . Resistant clones could be selected with a frequency ranging from 10(-7) to 10(-8) from the parent strain at twice the MIC . The following was observed from the study of six clinical isolates: (a) there was no uniform pattern of cytoplasmic membrane proteins among the isolates; (b) during the phase of growth, the number of high-molecular-weight proteins showed a decrease according to the duration of growth; (c) the addition of 1% w/v glucose to the growth medium resulted in the loss of an approximately 75 kD protein in five of the six strains; (d) subinhibitory concentrations of various antibiotics (1/4 of the MIC) resulted in major changes of cytoplasmic membrane proteins, when the strains were exposed to oxacillin, whereas this was not the case for erythromycin, rifampicin, gentamicin, trimethoprim, fleroxacin and vancomycin; (e) the resistant clones differed from their sensitive parent strains in their cytoplasmic membrane protein patterns . The alterations were not characteristic either for the strain or for the antibiotic considered, thus suggesting an 'unspecific' defense mechanism characterized by individual changes of the cytoplasmic membrane proteins. Scand J Infect Dis, 1992, 24(5), 607 - 12 Kinetics of serum levels of interleukin-6 in Staphylococcus aureus septicemia; Soderquist B et al.; The kinetics of Il-6 serum concentrations were analyzed in 17 patients with a culture verified Staphylococcus aureus septicemia . The analyses were performed using an antigen capture immunoassay . All patients had elevated IL-6 serum concentrations on admission . In most cases a subsequent rapid decrease to low levels was registered within 10 days . The IL-6 serum concentrations reflected the clinical course . In sera sampled on admission a relationship was found between IL-6 serum concentrations and levels of C-reactive protein. Miner Electrolyte Metab, 1992, 18(2-5), 174 - 85 Effect of malnutrition and uremia on impaired cellular host defence; Haag-Weber M et al.; There is a high incidence of infection in hospitalized patients with chronic renal failure . Nutritional and metabolic factors, as well as vitamins and trace elements are involved in impaired host defence and altered PMN function in dialysis patients . A circulating peptide (GIP) could be isolated from uremic serum that inhibits PMN glucose uptake, chemotaxis, oxidative metabolism and intracellular killing of Staphylococcus aureus . In addition to enhanced susceptibility to infection by impaired PMN function, uremic patients show profound defects of specific immune system represented by monocytes, B cells and T cells . T cells show decreased proliferation and Il-2 production on the one hand and enhanced Il-2 receptor expression on the other . Monocytes fail to elicit adequate help for T cell proliferation despite normal production of Il-1 and Il-6, but they produce elevated amounts of TNF alpha . B cells secrete decreased amounts of IgG and respond insufficiently to various vaccines . Malnutrition and uremia induce severe alterations of host defence and specific immune system if a combination of both these diseases occur. Zh Mikrobiol Epidemiol Immunobiol, 1992, (5-6), 4 - 7 {The effect of phospholipids on the adhesive properties of bacteria}; Tananova GV et al.; The present work shows that choline-containing phospholipids (lysophosphatidylcholine and lyso-1-alkyl-sn-glycerophosphocholine) inhibit the adhesion of some strains: Bacterium bifidum 1, B . adolescentis MC-42, B . longum B . 379M, Staphylococcus aureus P 209 and Klebsiella pneumoniae 52 . Phosphatidylcholine produces no effect on the adhesiveness of these strains, while platelet activation factor stimulates adhesiveness only in strain S . aureus 209 . The stimulating or inhibiting action of phospholipids on the adhesive process of microorganisms depends on the species of bacteria and on the concentration of reagents. Microbios, 1992, 71(287), 145 - 8 Inhibition of the electron transport system in Staphylococcus aureus by trimethylamine-N-oxide; Suzuki S et al.; Trimethylamine-N-oxide (TMAO) inhibited the growth of Staphylococcus aureus but not of S . epidermidis, which is the main flora in ripening squid, the Japanese traditional sea food Ika-shiokara . This selective inhibition of S . aureus was based on the inhibition of the electron transport system . The cytochrome fraction isolated from S . aureus was converted from the reduced to the oxidized form by the addition of TMAO . It is suggested that the inhibition occurred between cytochrome B and cytochrome o. Bull Soc Pathol Exot, 1992, 85(4), 271 - 5 {Staphylococcus aureus nosocomial infections in an intensive care milieu (1985-1989) in Tunis}; Ben Hassen A et al.; 221 strains of Staphylococcus aureus oxacillin resistant (MetiR) caused nosocomial infections were isolated from 1985 to 1989 in a medical intensive care unit . The survey of susceptibility to antibiotics was established according to the computerized data of disk susceptibility test . The resistance phenotypes to beta-lactams, aminoglycosides and macrolides were established for epidemiological study . S . aureus infections were mainly bacteraemia (31%) and peritonitis (12%) . These isolates were resistant to oxacillin with a high level (mean MIC 386 micrograms/ml) . Their resistance phenotypes were MLSBc (constitutive resistance to macrolides, lincosamine and streptogramines B) in 53% and S + KGT (resistance to streptomycin, kanamycin, gentamicin and tobramycin) in 61% . All the isolates were susceptible to pristinamycin and vancomycin (MIC 0.1 and 2 micrograms/ml) . These phenotypes related to the spread of multiply drug resistant strains were responsible of nosocomial outbreaks . Strains with the same pattern of resistance were isolated among the medical staff and in the environment . Infection control measures allowed to stop these outbreaks. Dermatology, 1992, 185(3), 175 - 80 Staphylococcus aureus leukocidin: a new virulence factor in cutaneous infections? An epidemiological and experimental study; Cribier B et al.; Panton-Valentine leukocidin (PVL) is a Staphylococcus aureus (SA) exotoxin, which kills human granulocytes and monocytes in vitro . Among 43 SA strains from cutaneous infections, 12 were PVL producers, whereas among 49 blood culture strains, only 1 produced PVL . Most PVL-producing strains (11/22) came from 22 primitive cutaneous infections, especially furuncles and abscesses, while only 1 PVL-producing strain came from 21 secondary infections of dermatoses such as bullous or pruritic diseases . Intradermal injections of PVL in rabbits induced edema, erythema and necrosis; histopathological changes at the injection sites consisted mainly in leukocytoclasis and vascular necrosis . All changes were dose dependent, and previous immunization of rabbits partially neutralized PVL-induced effects . Production of PVL in vivo after injections of bacteria seems to be low, and the histopathological lesions induced in the rabbit skin do not appear to be specifically related to PVL activity . However, PVL is a good candidate as a new virulence factor in cutaneous SA infections. Retina, 1992, 12(2), 108 - 12 Subretinal neovascularization secondary to choroidal septic metastasis from acute bacterial endocarditis; Munier F et al.; The clinical features of an infective embolic choroidopathy are described, from its early onset to late complications in a 45-year-old man with acute Staphylococcus aureus endocarditis of the aortic valve . Initial fundus examination revealed, in addition to fresh choroidal lesions, stigmata of a previous embolic episode secondary to endocarditis from Actinobacillus actinomycetemcomitans . The choroidal lesions were extremely asymmetrical, with a clear preference for localization in the left eye . Similar ocular findings were seen in a 78-year-old female with mitral valve prolapse and acute S . aureus endocarditis . In this case, however, choroidal involvement from septic emboli spread was bilateral and roughly symmetrical . Choroidal neovascular membranes arising in scars from choroidal septic emboli occurred in the macular area of the left eye of both patients, 10 months and 5 years after embolization, respectively. Int Orthop, 1992, 16(3), 237 - 9 Listeria monocytogenes infection in prosthetic joints; Allerberger F et al.; Listeria monocytogenes is increasingly recognised as a cause of prosthetic joint infection . These infections tend to be more insidious and indolent in contrast to similar infections with Staphylococcus aureus . They can occur in patients who are immunocompromised due to malignancy or other illness or in nonimmunocompromised elderly patients . Listeria infections should not be treated with cephalosporins and prolonged antimicrobial treatment is generally advised for prosthetic joint infections . We noted a high affinity of L . monocytogenes for foreign bodies . Three of twenty-seven cases 11%) of listeriosis under the period of investigation were associated with foreign bodies (hip prosthesis, knee prosthesis and arterial graft). Drugs Exp Clin Res, 1992, 18(3), 81 - 8 Effects of macrolides on ultrastructure of Staphylococcus aureus during postantibiotic phase; Watanabe T et al.; The postantibiotic effects (PAEs) of macrolide antibiotics, such as midecamycin acetate (Miocamycin, MOM), erythromycin (EM), josamycin (JM) and clarithromycin (CAM), on Staphylococcus aureus and the ultrastructure of the pathogen during the postantibiotic phase were investigated . After exposure to 2 x MIC for 2 h, MOM showed the longest PAE of 3.9 h, while EM, JM and CAM showed PAE durations 1.2, 2.5 and 1.9 h, respectively . On examining the serum levels of these agents in man, the longest PAE of 2.4 h was induced by exposure to MOM . JM and CAM induced PAEs for durations of 1.4 and 1.3 h, but EM hardly induced the PAE . The ultrastructure was examined by transmission electron microscopy, and thick cell walls with an undulating outer layer and a multiple thick cross-section were observed for 4 h after exposure to 2 x MIC of MOM for 2 h . After exposure to 2 x MIC of EM, JM and CAM for 2 h, ultrastructural changes were observed for 1, 2 and 2 h, respectively . The size of these cells was about 1.5 to 2 times larger than the normal cells . Ultrastructural changes in S . aureus were observed during the PAE phase of each macrolide. Adv Exp Med Biol, 1992, 319, 137 - 43 "Inverse" effects of cortisone in experimental infection of mice; Gillissen G; Hydrocortisone (HC) ameliorated the course of experimental infection with E . coli given intraperitoneally . In experiments with Staphylococcus aureus and treatment with endotoxin on the day of infection, HC had also a beneficial effect, however only when LPS was injected intraperitoneally and bacteria intravenously . With the footpad swelling reaction as a model for cellular immunity it could be shown that HC alone reduced the reactivity by 75 per cent . Under endotoxin treatment, however, HC induced further inhibition when LPS was given intraperitoneally the day of sensitization but changing to a stimulatory effect when LPS was given two days later . In case of intravenous application of LPS, HC had no effect. Ultrasound Med Biol, 1992, 18(6-7), 601 - 6 Conservation of bactericidal activity in ultrasound-exposed murine peritoneal phagocytic cells; Prohaska SS et al.; Murine peritoneal exudate cells (PEC), predominantly macrophages, were insonated in vitro with burst-mode ultrasound and assayed for their ability to phagocytose and kill Staphylococcus aureus . PEC were exposed at 37 degrees C in rotating tubes to 1-MHz, burst-mode (10 ms on, 10 ms off) ultrasound at 3.7 +/- 0.2 W/cm2 ISPTA (7.4 +/- 0.4 W/cm2 ISPBA) for 150 s . Bactericidal activity was assayed at 1, 2, and 3 h after exposure and subsequent 37 degrees C incubation with the bacteria for 20 min . In these experiments, which comprised 17 treated and 7 sham-treated control samples, there was no significant difference in results between treated and control samples (p > 0.29). Rev Med Chir Soc Med Nat Iasi, 1992 Jan-Jun, 96(1-2), 19 - 25 {Acute nosocomial bacterial meningitis . Apropos 27 cases}; Dimitriu SM et al.; Our series includes 27 patients with nosocomial meningitis (22 post neurosurgery and 5 post spinal puncture) of whom 15 with stated etiology (gram-negative bacilli and staphylococcus aureus in equal shares) . Under the treatment with chloramphenicol + rifampicin + gentamicin a cure was obtained in 22 cases, improvement in 3 cases and 2 patients died. Nephrologie, 1992, 13(4), 171 - 7 Peritonitis prevention in continuous ambulatory peritoneal dialysis; Luzar MA; Although peritonitis remains the major cause of morbidity in CAPD, peritonitis rates are declining in European and other countries . This article reviews approaches that are both decisive and promising concerning the prevention of peritonitis in CAPD . Clinical results with both reusable and single-use Y sets are discussed . These systems appear to have a significant impact on the reduction of intraluminal contamination, particularly Staphylococcus epidermidis . The importance of the flush-before-fill technique is reviewed in the context of the new disposable Y sets . In vitro studies confirm that 100 mls of fresh dialysate flushed from the new bag to the drainage bag at the appropriate time during the exchange can eliminate microorganisms that do not possess adherence factors, providing long periods of incubation are not encountered . Future prevention measures for the reduction of Staphylococcus aureus peritonitis are discussed in light of evidence identifying pre-CAPD nasal carriers as high risk patients for subsequent exit-site infection and S . aureus peritonitis . These measures include methods such as the application of antibiotics such as mupirocin to the anterior nares before and during CAPD . The roles of intraperitoneal IgG therapY and staphylococcal vaccination as additional therapeutic approaches to infection control in peritoneal dialysis are also discussed. Int Arch Allergy Immunol, 1992, 98(4), 339 - 42 Differential increase in 12-HETE release and CD29/CD49f expression of platelets from normal donors and from patients with atopic dermatitis by Staphylococcus aureus; Neuber K et al.; The generation of the arachidonic acid-derived inflammatory mediator 12-hydroeicosatetraenoic acid (HETE) and the expression of CD29 as well as CD49f from unstimulated and stimulated platelets has been studied in patients with atopic dermatitis (AD) as well as in healthy volunteers . Heat-killed clinical isolates of Staphylococcus aureus served as stimuli . Unstimulated platelets from patients with AD produced higher amounts of 12-HETE compared to platelets from normal donors . The absolute 12-HETE release from platelets of patients with AD was significantly higher compared to the control group after stimulation with heat-killed S . aureus, whereas the relative increase remained . The expression of CD29 and CD49f on unstimulated platelets of patients with AD was markedly enhanced compared to platelets from normal donors . Stimulation with S . aureus led to similar results as to the CD29 expression on normal and atopic platelets or to a markedly higher expression of CD49f on platelets from normal donors . As compared to platelets from normal donors the CD49f expression on atopic platelets was slightly enhanced by S . aureus . Our data emphasize that platelets may play an important role in the pathogenesis of AD by an increased preactivation and by an enhanced responsiveness to S . aureus which colonizes permanently the skin of patients with AD. Clin Neurol Neurosurg, 1992, 94(3), 253 - 5 Iatrogenic acute spinal epidural abscess with septic meningitis: MR findings; Shintani S et al.; A contaminated catheter used in epidural anesthesia in a 71-year-old female produced acute epidural abscess and septic meningitis . Methicillin-resistant Staphylococcus aureus (MRSA) was detected in a culture of the epidural pus . Both T1- and T2-weighted MR images showed low intensity mass lesion compressing the thecal sac behind the vertebral body L3 . The low intensity lesion was probably pus with gas component . In these low intensity lesions in MR findings with gas component, MR was superior to myelography because it visualized both the degree of compression to the thecal sac and extension of the lesion in all directions. Vet Res Commun, 1992, 16(2), 107 - 15 Polymorphonuclear neutrophil leukocyte function in clinical bovine patients and in cows with or without Staphylococcus aureus mastitis; Anderson KL et al.; A fluorochrome microassay was used to investigate peripheral blood polymorphonuclear leukocyte (PMNL) function in cattle . Glass-adherent PMNL were reacted with Staphylococcus aureus preincubated in 20% bovine serum for 30, 60 and 90 min . Coverslips were stained with acridine orange (AO) followed by crystal violet to quench extracellular bacterial fluorescence . PMNL function was evaluated by counting the number of dead (stained red with AO) and live (stained green with AO) S . aureus contained within 100 PMNL . A phagocytic index was calculated as the average number of bacteria contained within PMNL . The percentage killing of S . aureus was calculated from the average proportion of S . aureus within PMNL that were dead . Six clinically normal Holstein calves, 3-4 months of age, were sampled on 6 consecutive days . PMNL phagocytosis and killing did not vary significantly (p greater than 0.05) among repeated samplings per calf . PMNL function increased with increasing time of incubation of PMNL with S . aureus . Means (+/- SD) for percentage killing were 46.7 +/- 13.1, 57.4 +/- 11.6, and 62.1 +/- 9.8% for 30, 60 and 90 min of reaction, respectively . Means (+/- SD) for the phagocytic index were 2.9 +/- 0.8, 3.6 +/- 1.0, and 4.2 +/- 1.1 bacteria/PMNL for 30, 60 and 90 min of reaction, respectively . PMNL function was determined in 30 normal cattle of various breeds, age and sex, and these values were pooled to provide normal values for PMNL function . When values for bovine clinical patients (n = 25) with various diagnoses were compared with normal values (defined by the mean +/- 2SD for the 30 normal cattle) for PMNL function, only one patient was observed to exhibit PMNL hypofunction . A cow with disseminated intravascular coagulation in association with peracute coliform mastitis exhibited decreased PMNL killing capacity . Abnormal PMNL function was uncommon in the hospital population studied . Peripheral blood PMNL function was evaluated in lactating Holstein cows with (n = 15) or without (n = 15) chronic subclinical S . aureus mastitis . There was no significant (p greater than 0.05) difference in PMNL function among these cows. Avian Dis, 1992 Jan-Mar, 36(1), 149 - 53 A coagglutination assay with monoclonal antibodies for rapid laboratory identification of Mycoplasma synoviae; Morsy MA et al.; An agglutinating monoclonal antibody (MAb S2) specific for a 55,000-molecular-weight surface protein of Mycoplasma synoviae was developed by fusion of spleen cells from immunized BALB/c mice with P3X63 Ag8.653 myeloma cells . Immunogold labeling experiments confirmed the cell surface location of the MAb-recognized antigen . MAb S2-coated Staphylococcus aureus was used in a rapid slide coagglutination assay . Eleven M . synoviae strains, including the type strain WVU 1853, coagglutinated with MAb-coated S . aureus, but five M . gallisepticum strains (PG31, S6, R, F, and A5969) did not. Cytometry, 1992, 13(1), 83 - 9 Detection of granulocyte reactive oxygen species formation in whole blood using flow cytometry; Himmelfarb J et al.; We have developed a technique for analysis of granulocyte reactive oxygen species formation in whole blood using flow cytometry and two color immunofluorescence . This technique relies upon the use of specific fluorescent dye (LDS-751) to stain nucleated cells, eliminating erythrocytes from analysis . Using LDS-751, forward angle light scatter, and 90 degrees side scatter, a granulocyte gate, monocyte gate, and lymphocyte gate were identified . Analysis with multiple FITC conjugated monoclonal antibodies demonstrated greater than 95% purity of a flow cytometrically identified granulocyte population in whole blood without physical manipulation of the blood . Utilizing 2'7' dichlorofluorescein diacetate (DCFH-DA), we were able to measure granulocyte intracellular reactive oxygen species production . Dose response curves were obtained for the effect of granulocyte agonists phorbol myristate acetate, FMLP, and heat fixed Staphylococcus aureus on reactive oxygen species production . The techniques described in this paper should be useful for measuring granulocyte activation in vivo with flow cytometry. Comp Immunol Microbiol Infect Dis, 1992 Jan, 15(1), 41 - 6 Systemic profiles of antigen-specific lymphocytes in animals chronically exposed to staphylococcus antigen in the mammary region; Dobrzanski MJ et al.; Systemic profiles of lymphocytes were assessed in goats exposed chronically with Staphylococcus antigens in the supramammary region . Animals were inoculated three times subcutaneously in the right supramammary region with heat-killed Staphylococcus aureus antigen (HKS) at 1 month intervals . Prior to immunization and 1 week following each injection, 3 and 6 day cultures of peripheral blood mononuclear cells were made to determine proliferative responses of lymphocytes to HKS and the polyclonal T cell mitogen phytohemagglutinin (PHA) . Peripheral blood lymphocytes responded significantly to both HKS and PHA in 3 day cultures after the second injection and showed peak responses after the final immunization, suggesting that repeated local injection of S . aureus antigen at the supramammary region, can induce an anamnestic response to the antigen in the peripheral blood of these animals with a concomitant increase in the responsiveness to the polyclonal mitogen, PHA . In contrast, initial antigen challenge induced little, if any, increase in responses to the specific antigen or mitogen when compared to pre-injection states . These data may also suggest that non-reactivity of peripheral blood lymphocytes to the HKS antigen immediately after the primary injection of antigen may be the result of local retention of antigen-reactive cells at the sites of infection. AIDS, 1992 Jan, 6(1), 55 - 63 HIV-induced, HIV-specific in vitro antibody response by B-cells from HIV-seropositive individuals; Delfraissy JF et al.; OBJECTIVE: Recent studies have shown that B-cells from HIV-infected patients can secrete anti-HIV antibodies in vitro and that they represent 20-40% of immunoglobulin (Ig)-secreting B-cells in vivo . This study was designed to investigate the precise role of HIV in this in vitro antibody production . DESIGN AND METHODS: B-cells from HIV-infected patients {asymptomatic, n = 28; symptomatic (AIDS), n = 14}, from seronegative adult volunteers (n = 22) and subjects at high risk for HIV infection (n = 15) were cultured in vitro in the presence of pokeweed mitogen, Staphylococcus aureus cowan or HIV, and T-cells or interleukins (IL) . Non-specific Ig production and specific anti-HIV antibody (Ab) production were measured by enzyme-linked immunosorbent and Western blot assays . RESULTS: We found that HIV induced a specific response in cultured B-cells from seropositive patients, in contrast with cultured B-cells from uninfected normal individuals . The characteristics of the HIV-induced response differed from those of a spontaneous or a mitogen-induced response . Anti-HIV Ab production was optimal on day 8-10, when B-cells were cultured with recombinant IL-2 and recombinant interferon-alpha in the presence of infectious virus or recombinant gp160 Env protein . The anti-HIV Ab were mainly directed against Env proteins . Interaction of HIV with B-cells involved surface IgG but not CD4 antigen . Autologous CD8+ T-cells had a non-specific inhibitory effect . Both CD5+ and CD5- B-cells produced anti-HIV Ab . No anti-HIV Ab production was observed in B-cells from high-risk HIV-seronegative individuals . CONCLUSION: HIV (infectious virus or gp160) can induce B-cells from infected patients to secrete specific anti-HIV Ab in vitro. Immunology, 1992 Jan, 75(1), 136 - 42 Effects of FK506 and cyclosporin A on cytokine production studied in vitro at a single-cell level; Andersson J et al.; Mononuclear cells obtained from human blood were mitogen or antigen activated in vitro in the presence or absence of FK506 or cyclosporin A (CsA) . Cytokine production was studied at a single-cell level by ultraviolet (UV) microscopy of fixed permeabilized cells using cytokine-specific monoclonal antibodies (mAb) . Phenotypic characterization of the monokine-producing cells was achieved by two-colour immunofluorescent staining . Cytokine production after antigen activation with Staphylococcus aureus enterotoxin A (SEA) was significantly reduced . FK506 or CsA inhibited SEA-induced tumour necrosis factor-alpha (TNF-alpha) production both in monocytes (P less than 0.01) and in lymphocytes (P less than 0.001), at a drug concentration of 1-25 ng/ml for FK506 and 100-500 ng/ml for CsA . Lymphocyte synthesis of interleukin-2 (IL-2), interferon-gamma (IFN-gamma) and TNF-beta after SEA activation was also significantly reduced by either of the drugs . In contrast, endotoxin-induced monokine production (TNF-alpha and IL-6) after lipopolysaccharide (LPS) stimulation was unaffected by FK506 or CsA even when added in concentrations as high as 1000 ng/ml . When the cells were stimulated by phorbol ester (phorbol 12-myristate 13-acetate, PMA) plus calcium ionophore (ionomycin), FK506 and CsA inhibited, in a dose-dependent manner, the production of IL-2, IL-4, IL-5, IFN-gamma and TNF-alpha . The 50% inhibitory concentration (IC50) for FK506 or CsA on the cellular synthesis of the various cytokines varied between 0.6 and 1.0 ng/ml and 20 and 60 ng/ml, respectively . Further stimulation by addition of anti-CD28 mAb to the cultures resulted in an augmented IL-2 and IFN-gamma production which was resistant to both FK506 and CsA . This report delineates extensive similarities between the two drugs in mechanisms of immunosuppression by blockade of identical interleukin production . Depending on the mode of cell activation the two drugs inhibited not only cytokine production in lymphocytes but also antigen-induced monokine (TNF-alpha) production in macrophages, although the optimal immunomodulatory effect of FK506 was achieved at a concentration approximately 50-fold lower than that of CsA. Appl Microbiol Biotechnol, 1992 Jan, 36(4), 483 - 6 Production of recombinant human glucagon in the form of a fusion protein in Escherichia coli; recovery of glucagon by sequence-specific digestion; Ishizaki J et al.; Recombinant human glucagon was successfully produced with a high level of expression in Escherichia coli as a fusion protein with human interferon gamma . The synthetic gene was designed to release glucagon, which does not contain glutamic acid residues, from fusion protein with the Staphylococcus aureus strain V8 protease that specifically cleaves the peptide bond on the carboxyl side of the glutamic acid residue . The resulting glucagon was purified to homogeneity by a combination of C18 reverse-phase HPLC and ion-exchange HPLC . The yield of intact glucagon obtained from 11 of culture was approximately 12 mg . The structure of recombinant human glucagon was confirmed by HPLC and amino acid composition/sequence analyses. Acta Univ Palacki Olomuc Fac Med, 1992, 134, 7 - 9 Effects of static magnetic field on some pathogenic microorganisms; Grosman Z et al.; A high power superconductive magnetostatic system MAGNEX was used by the authors for two microbial strains Escherichia Coli and Staphylococcus Aureus, the magnetic induction values were 0.5-4 T and the exposure time was 30-120 minutes . In this paper morphological traits, standard biochemical tests, sensitivity and effects influence of some chosen sorts of antibiotics are considered and evaluated here . All the above given criteria and facts were analysed and the authors have come to the conclusion that magnetic DC field has no significant influence on the growth, the biochemical activity as well as on the antibiotics effects on the strains examined. Adv Perit Dial, 1992, 8, 283 - 7 A new catheter to prevent exit-site infection in peritoneal dialysis; Catizone L et al.; In order to prevent exit-site infection, we studied a new Tenckhoff-derived catheter, named the "Malpighi catheter," capable of avoiding sinus tract formation . The outer cuff of this new device is 3.5 cm long and is deliberately positioned half-extruded; in fact, half of the cuff remains outside the skin exit-site . The implantation technique is identical to that of the standard two-cuff Tenckhoff catheter . We implanted eight Malpighi catheters in 5 CAPD and 3 IPD patients . The observation period was 146 patient-months (range 14-23, M +/- SD 18.2 +/- 3.3) . We observed excellent adhesion between the outer cuff and surrounding tissue . Actually, by pulling the catheter the skin around the half-extruded cuff becomes cone-shaped, with the cone's apex tightly stuck to cuff and the sinus tract disappearing completely . Only one case of exit-site infection by Staphylococcus aureus and two cases of ulcer of the skin beneath the external part of the half-extruded cuff were observed . These complications were resolved completely . No catheter needed to be removed and there were no leakages . The histological study of the cuff showed a good infiltration of the dacron cuff by fibrous tissue . On the grounds of our preliminary experience, we believe that the absence of the sinus tract, the formation of an efficient mechanical and bacterial barrier and the reduction of exit-site infection incidence are all factors that encourage further research. Adv Perit Dial, 1992, 8, 265 - 8 Does catheter immobilization reduce exit-site infections in CAPD patients? Turner K, Edgar D, Hair M, Uttley L, Sternland R, Hunt L, Gokal R. This study was undertaken to ascertain the effectiveness of an immobilization device in reducing exit site infections (ESI) in CAPD patients, and whether immuno-suppressive therapy, diabetes, disconnect system and Staphylococcus aureus nasal carriage had any bearing on the incidence of ESI . Sixty-six patients having a Tenckhoff catheter placement were randomly allocated into one of three groups; immobilizer, tape and non-immobilized group . The groups were monitored for the incidence of ESI over a total period of 347 patient months . The results show no significant difference in infection rates between the three groups, nor do the factors mentioned have any bearing on ESI rate . Whilst immobilization is important, the ineffectiveness of this device was probably related to its design problem. Adv Perit Dial, 1992, 8, 253 - 7 Staphylococcus aureus nasal carrier status (SANCS) in CAPD patients; is it induced or favored by subcutaneous rHu-erythropoietin? Selgas R, Castro MJ, Bajo MA, Fernandez-Reyes MJ, Lopez-Revuelta K, Romero JR, de Alvaro F, Borrego F, Rinon C. Staphylococcus aureus nasal carriage status (SANCS) has been recognized as a risk factor for patients on CAPD, due to a higher probability of suffering peritoneal catheter infections . The use of subcutaneous drugs (insulin dependent diabetics, drug addicts, HD patients and antiallergic vaccines), has been associated with increased risk of SANCS . On CAPD, erythropoietin (EPO) is almost universally used by the subcutaneous route . The objective of this paper was to evaluate the incidence and prevalence of SANCS in 85 CAPD patients by means of nasal smear and the influence of SANCS on peritoneal and catheter infection rate . Patients were divided in four groups according to diabetic status and EPO treatment (mean dose 2000 u . twice a week) . The prevalence of SANCS in control groups was 30% in non-diabetics and 23% in diabetics . EPO treated patients showed a prevalence of SANCS of 39% in non-diabetics and 45% in diabetics due to the presence of 7 and 5 carrier patients respectively . SANCS patients (29% of the population), suffered 45% of peritonitis and 42% of exit-site infections caused by S . aureus . In a prospective part of the study, there was no difference in the frequency of developing positive cultures among EPO and control (30% of patients) . No male EPO treated patients developed SANCS . We conclude that it is necessary to monitor S . aureus nasal carrier status periodically in CAPD patients especially in women . Whether or not subcutaneous erythropoietin treatment is implicated pathogenetically with SANCS, is not clarified by our data because of the frequent spontaneous appearance of SANCS among CAPD patients.(ABSTRACT TRUNCATED AT 250 WORDS) Adv Perit Dial, 1992, 8, 242 - 5 Treatment of Staphylococcus aureus nasal carriers in CAPD with mupirocin; Perez-Fontan M et al.; We have studied the efficacy of topical Mupirocin for elimination of Staphylococcus aureus (Staph . aureus) nasal carriage in CAPD patients . Staph . aureus nasal carriers in our CAPD program were randomized to one of two groups: Group 1, treated with Mupirocin, and Group 2, treated with neomycin sulphate nasal ointment . The prevalence of Staph . aureus nasal colonization was 44% for patients (24/54) and 17% for dialysis partners (5/29) . Group 1 included 11 patients and 1 partner, and Group 2, 8 patients and 2 partners . In Group 1, the eradication rate was 100%, and the recolonization rate was 0, 8, 41, 55 and 66% at 1, 2, 3, 6 and 10 months . In Group 2, the eradication rate was 40%, with a recolonization rate of 0.25 and 75% at 1, 2 and 3 months . Re-treatment with mupirocin was successful in 66% of the cases, compared to 20% for neomycin . The MIC90 of mupirocin for Staph . aureus was 0.5 mcg/mL, with an increase to 4 mcg/mL towards the end of the study . During the study period, there was a very low incidence of Staph . aureus peritonitis or catheter-related infections in patients treated with mupirocin . Secondary effects of mupirocin were negligible . Mupirocin is more effective than neomycin sulphate for the elimination of Staph . aureus nasal colonization in patients undergoing CAPD . Periodic re-treatment is frequently necessary, given the significant recolonization rate. Adv Perit Dial, 1992, 8, 219 - 22 Effect of glucose concentration and dwell time of dialysis fluid on the antibacterial defense in the peritoneal cavity of rats; Calame W et al.; To study the influence of dialysis fluid on the antibacterial defense in the peritoneal cavity of rats, especially glucose concentration and dwell time, an experimental infection was developed . Rats were injected intraperitoneally with dialysis fluid with a glucose concentration of 1.36%, 2.27%, or 3.86% or physiological saline . Subsequently 1, 4 or 18 hr thereafter an inoculum of approximately 3 x 10(8) colony forming units of Staphylococcus aureus was administered intraperitoneally (i.p.) Next, 24 hr after the inoculation the number of viable bacteria was determined . Peritoneal cells (PC) isolated 1, 4 and 18 hr after the administration of dialysis fluid were tested for their capacity to kill S . aureus in vitro . A positive relation was observed between the glucose concentration and the number of bacteria isolated; the longer the dwell time the lower this number . In vitro PC isolated at the various intervals did not differ in their capacity to kill S . aureus . It is concluded that the glucose concentration in dialysis fluid impairs the antistaphylococcal defense in the peritoneal cavity of rats . A relatively long dwell time enhances this defense . These results cannot be explained by a lower capacity of the PC to kill S . aureus in vitro. J Hosp Infect, 1992 Jan, 20(1), 25 - 34 Spread of Staphylococcus aureus strains of phage-type 95 in Denmark 1968-1989; Schonheyder H et al.; The spread of Staphylococcus aureus strains of phage-type 95 was traced retrospectively in Denmark by the review of more than 15,000 S . aureus bacteraemia isolates (1957-88) and from data collected by phage-typing of c . 260,000 isolates from all body sites (1977-89) . The first two type 95 strains had been isolated from blood in 1968, and after an interval of 3 years there was a steady increase of bacteraemia strains all over Denmark . From 1977 to 1989 the incidence of type 95 strains among isolates from all body sites increased from 3.8 to 18.8% . Different patterns of increase were recorded in 13 major hospitals and in various clinical departments of two hospitals and these were further analysed . Conjunctival swabs gave the highest percentage of type 95 strains and those from abscesses gave the lowest percentage . Of the type 95 bacteraemia strains 90.4% were resistant to penicillin, but neither methicillin nor gentamicin resistance was recorded. Agressologie, 1992, 33(4), 188 - 90 {Our experience of infectious risk in prosthetic breast surgery}; Herman D et al.; This retrospective clinical study was made to help determine the signs of infectious or healing problems in candidates for prosthetic breast surgery . It concerns 80 patients with 108 breast implants (44 breast reconstructions, 21 cases of hypoplasia, 8 breast asymmetric, 20 secondary procedures, 15 of them being changes of expansion implants by definitive implants) . Thirty one infectious complications (28.7%) were noted . In 22 cases it consisted of a resolutive hyperthermia . Nine cases of local complications were noted, with four implant exposures (3%) . Bacteriological samples were analyzed and were positive in four out of nine (Staphylococcus aureus) . In 8 out of the 9 cases the initial procedure consisted in a breast reconstruction . All 8 patients underwent postoperative radiotherapy after the initial mastectomy (Patey) . The implant type (implantable, pre-filled, expansion prosthesis), its location (retromuscular), the type of surgery (asymmetrical breast musculocutaneous or fasciocutaneous flap) don't seem to be responsible in the genesis of the complications . This retrospective study could not help determine which protocol of antistaphylococcal antibiotherapy suits lest . Infectious problems arose with or without antibiotics. Agressologie, 1992, 33 Spec No 3, 154 - 6 {Bacteriological relationship between organ donor and recipient . A survey apropos of 100 brain dead patients}; Voltz C et al.; The aim of the study was to evaluate the relationship between germs found in samples systematically taken in the organ donors and germs found in the corresponding recipients in the 8 post-operative days . The 100 brain-dead patients received oxacillin (2g every 4 hours) when the bacteriological samples were taken . 41% of the donors were germ carriers . The germs were mainly located in the respiratory track (Staphylococcus aureus meti-S (34%) and Hemophilus (29%)) . Among the 86 patients harvested, 40 donors were germ carriers and gave 132 organs, 46 donors were not germ carrier and gave 151 organs . The comparison between these two groups showed no difference . Assessment of infection occurring in organ recipient in the 8 post-operative days (germ, location, evolution) showed no difference, whether organs were removed from germ carrier donors or no . Comparison between the germs found in germ carrier donors and those found in recipients with sepsis showed a similitude in three cases (2.2%) . In the other cases, there is no relationship between the germs found in the donors and post-operative sepsis in the recipient. Drugs Exp Clin Res, 1992, 18(9), 367 - 9 In-vitro activity of daptomycin against a worldwide collection of methicillin-resistant Staphylococcus aureus; Brumfitt W et al.; Minimum inhibitory and minimum bactericidal concentrations (MIC and MBC) of daptomycin and of vancomycin have been compared against 80 strains of methicillin-resistant Staphylococcus aureus isolated from many parts of the world . In the presence of 30 mg/l of Ca++, daptomycin at 4 mg/l killed all the strains tested, and was only slightly less active than vancomycin. Pediatrie, 1992, 47(7-8), 531 - 4 {Non traumatic acute suppurative mediastinitis}; Varlet F et al.; Acute mediastinitis is uncommon . When it occurs, it usually follows an esophageal perforation or thoracic surgery . We report on a case of a 2-year-old girl with non traumatic mediastinitis secondary to a pharyngitis, due to Staphylococcus aureus . The anatomic pathways that connect the cervical region with the mediastinum explains how the infection can spread within the cervicothoracic spaces . Knowledge of this anatomy is also important in evaluating the possible causes of symptoms and signs in this area, and the possible complications of infections in these regions . The treatment associates antibiotics and drainage of the abscess by either surgical procedure or computed tomography guided selective needle aspirations. Cytometry, 1992, 13(5), 525 - 31 Dose effects of LPS on neutrophils in a whole blood flow cytometric assay of phagocytosis and oxidative burst; Bohmer RH et al.; Whole blood phagocytosis (P) and oxidative burst (OB), a rapid and sensitive flow cytometric method for quantifying neutrophil activation, was modified for single laser systems by using propidium iodide labeled Staphylococcus aureus and 2',7' dichlorofluorescein diacetate . The purpose of the present study was to characterize this assay with respect to the stimulatory activity of bacterial lipopolysaccharide (LPS) on phagocytosis . Blood from healthy donors was preincubated with log doses of bacterial LPS B (0.1-1,000 ng/ml) or sterile pyrogen-free saline at 37 degrees C from 0-120 minutes . LPS increased both P and OB in a dose-dependent manner (up to 62 and 121%, respectively) at all time points tested, and this effect on P and OB could be detected even with no preincubation . This LPS-induced phagocytic activity could be blocked by the addition of polymyxin B (10 micrograms/ml) during preincubation . The priming effect of LPS was maximal at 45 min . P and OB were inhibited by preincubation with EDTA at doses greater than 2 mM (60 and 80% inhibition, respectively) . These observations are consistent with the exquisite sensitivity of the neutrophil to endotoxin . This method can evaluate neutrophil response to immunomodulatory and chemotherapeutic agents in a physiological milieu . These findings re-emphasize the necessity of using pyrogen-free reagents in any study of neutrophil function. Antimicrob Agents Chemother, 1992 Jan, 36(1), 10 - 6 Potentiation of antibacterial activity of azithromycin and other macrolides by normal human serum; Pruul H et al.; The interaction of azithromycin with normal human serum was examined in relation to serum protein binding, MIC, and kinetics of killing of bacteria . While the binding of azithromycin to serum proteins is low (8.5% at a concentration of 0.01 mM in 95% serum), the presence of 40% serum during the MIC test decreased MICs by 26-fold for serum-resistant Escherichia coli and 15-fold for Staphylococcus aureus . Erythromycin had a similar but lesser effect, while roxithromycin was less active against S . aureus in the presence of serum . The rate of killing of E . coli and S . aureus by azithromycin was increased in the presence of serum . The enhancement of antibiotic activity by serum was pH independent, and heat inactivation and preabsorption with homologous bacteria failed to inhibit enhancement by serum . The macromolecular incorporation of {3H}thymidine by E . coli continuously exposed to 2 micrograms of azithromycin per ml (0.25x the MIC) and 40% serum was decreased by 80% at pH 7.8 and by 48% at pH 7.2, while azithromycin alone failed to inhibit incorporation . Inhibition of nucleic acid biosynthesis at pH 7.2 in the presence of serum was also detected with sub-MICs of erythromycin, norfloxacin, and gentamicin but not roxithromycin . A diffusible serum factor was shown to interact with azithromycin to inhibit the growth of E . coli in an agar diffusion assay to detect antibiotic-serum synergy. Eur J Clin Microbiol Infect Dis, 1992 Jan, 11(1), 55 - 8 In vitro activity of sparfloxacin and three other fluoroquinolones against methicillin-resistant Staphylococcus aureus and Staphylococcus epidermidis; Smith KR et al.; Fluoroquinolones are variably active against methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-resistant Staphylococcus epidermidis (MRSE) . The purpose of this study was to test the in vitro susceptibility of 50 isolates each of MRSA and MRSE to four of the new fluoroquinolones--sparfloxacin, ciprofloxacin, ofloxacin and norfloxacin--and to see if resistance could be induced in five susceptible strains of each species by serial passages with increasing concentrations of drug . A standard microdilution technique was used to determine minimal inhibitory concentrations (MICs) and minimal bactericidal concentrations (MBCs) of each drug for each isolate . Agar dilution plates were also prepared containing concentrations of drug varying from one-half of the reported MIC to 128 times the reported MIC, and microorganisms persisting were serially passaged . Initially, 98% of the strains of MRSA were susceptible to the fluoroquinolones . MBCs were essentially identical to MICs . Similarly, 96% of the strains of MRSE were susceptible . Following exposure to increasing concentrations of each fluoroquinolone, resistance appeared to emerge less rapidly to sparfloxacin and ofloxacin than to norfloxacin and ciprofloxacin. Infect Immun, 1992 Jan, 60(1), 202 - 5 Pertussis toxin partially inhibits phagocytosis of immunoglobulin G-opsonized Staphylococcus aureus by human granulocytes but does not affect intracellular killing; Hiemstra PS et al.; The aim of the present study was to determine whether pertussis toxin (PT)-sensitive GTP-binding proteins (G proteins) are involved in the signal transduction pathway(s) used for phagocytosis and intracellular killing of bacteria by human granulocytes . Treatment of granulocytes with PT resulted in decreased phagocytosis of immunoglobulin G (IgG)-opsonized Staphylococcus aureus but did not affect subsequent intracellular killing of these bacteria . PT also caused a decrease in the extracellular release of superoxide anion (O2-) and hydrogen peroxide (H2O2) by granulocytes in response to S . aureus opsonized by IgG . However, neither the phagocytosis nor the intracellular killing of S . aureus opsonized by fresh serum was affected by PT, and the release of O2- was partially inhibited . The release of O2- in response to serum-treated zymosan, opsonized mainly by complement components, was also only partially inhibited by PT . It is therefore possible that PT inhibits responses mediated through complement receptors to a lesser extent than those mediated via Fc gamma receptors . The results of this study indicate that PT-sensitive G proteins are involved in the signal transduction pathways that mediate the phagocytosis of IgG-opsonized bacteria and the accompanying respiratory burst. Nurs Pract, 1992, 5(2), 2 - 7 Methicillin resistant Staphylococcus aureus and its implications for nursing practice: a literature review; Simpson S; The subject of methicillin-resistant Staphylococcus aureus (MRSA) and its implications for nursing practice are reviewed . Resistance of Staphylococcus aureus to antibiotics, and the emergence and incidence of MRSA both in the United Kingdom and abroad, are examined . Modes of transmission of MRSA by transfer of affected patients and, specifically, cross infection by staff are reviewed with methods to prevent cross infection, including screening of staff, handwashing techniques, barrier and cohort nursing, being examined . The costs of an MRSA outbreak, both financially and psychologically, to staff and patients are discussed . Guidelines for the control of epidemic MRSA are explained in the context of the above topics, and all areas are related to improving nursing practice. Drugs Exp Clin Res, 1992, 18(10), 415 - 22 Efficacy of subinhibitory concentration of pefloxacin in preventing experimental Staphylococcus aureus foreign body infection in mice; Carsenti-Etesse H et al.; Adhesion is the first step leading to colonization and infection of a foreign body (FBI) . To assess the ability of a subinhibitory concentration (subMIC) of pefloxacin (P) to prevent such infection, an experimental model was developed in Swiss albino mice . Subcuts of polyurethane catheters (Vygon) were placed in the peritoneal cavity of animals and 24 hours later, different inocula of an adherent strain of Staphylococcus aureus (SA) (MIC of P:0.8 mg/l) were injected i.p . Unexposed SA served as controls . Two days later the removed catheters, blood and spleen specimens were quantitatively cultured for bacterial content and identity . Infection was defined as more than 10 CFU/ml of SA recovered . Significant protection of mice, with lower dissemination, was found with inoculum sizes of 10(5) and 10(6) . These results suggest that subMICs of P may confer protection against foreign body infection. Med Dosw Mikrobiol, 1992, 44(3-4), 93 - 6 {Evaluation of susceptibility to antibiotics of Staphylococcus aureus strains resistant to methicillin}; Trzcinski K et al.; Methicillin-resistant strains of Staphylococcus aureus (MRSA) constitute a serious diagnostic and therapeutic problem . Over 500 strains of Staphylococcus aureus were tested for susceptibility to methicillin . By application of a screening method, 13.7% of these strains were classified as methicillin-resistant . Over 95% of these strains were isolated from hospital infections . Applying criteria of belonging of these strains to methicillin-resistance classes it was found that 49.3% belonged to class II, 31.2% to class III and 19.5% to class IV . Analysis of susceptibility to antibiotics of MRSA strains demonstrated significant differences between class II and between class III and IV in resistance to imipenem, gentamycin, erythromycin and tetracycline . All tested strains were susceptible to ciprofloxacin, ofloxacin, vancomycin and teicoplanin . The screening method (25 mg methicillin/l of TSA medium) results in obtaining of univocal results of determination of methicillin-resistance in S . aureus. Ginekol Pol, 1992, 63(7), 337 - 43 {Use of vancomycin in treating staphylococcal septicemia in newborns}; Godula-Stuglik U et al.; The authors report the results of vancomycin treatment of staphylococcal septicemia in 26 newborns, including 17 prematures and 9 full-term . It was found, that all coagulase-negative strains and 91% of Staphylococcus aureus were sensitive to this antibiotic . Complete cure in 65% and clinical improvement in 10% were obtained . Death (12%) in very low birth weight prematures were due to secondary complications of septicemia, congenital malformations and intracerebral haemorrhages . Administration of vancomycin in generally accepted doses caused no side effects and is worth in newborns, in which other antibiotics has been given unsuccessfully. Zh Mikrobiol Epidemiol Immunobiol, 1992, (11-12), 15 - 6 {The production of toxic shock syndrome toxins and enterotoxins by Staphylococcus aureus}; Petras P et al.; The production of toxins of the toxic shock syndrome and enterotoxins by S . aureus of clinical origin was studied by means of commercial standard antisera and standard toxins . The study revealed that among 511 strains isolated in Russia toxigenic ones constituted 41%, and among 592 strains isolated in Czechia toxigenic ones constituted 63% . The capacity for producing toxins may be used as an epidemiological marker. Autoimmunity, 1992, 14(1), 23 - 32 Human B cell differentiation induced by microbial superantigens: unselected peripheral blood lymphocytes secrete polyclonal immunoglobulin in response to Mycoplasma arthritidis mitogen; Crow MK et al.; Microbial superantigens (SA) activate a significant portion of the T cell repertoire based on their dual avidity for MHC class II antigens and T cell receptor (TCR) epitopes common to products of one or several TCR beta chain variable gene families . While SA that induce massive T cell proliferation and cytokine secretion have been implicated in clinical syndromes characterized by shock and generalized immunosuppression, SA activation of a more restricted T cell response may also have significant, perhaps immunostimulatory, effects on the immune system . To investigate this issue, we measured 3H-thymidine incorporation and polyclonal IgM and IgG secretion by normal human peripheral blood mononuclear cells (PBMC) cultured with a panel of microbial SA, including the Staphylococcus aureus-derived SA, SEA, SEB, SEC-1, SEC-2, SEC-3, SEE, TSST-1, and the Mycoplasma arthritidis-derived SA, MAM . The S . aureus-derived SA induce vigorous proliferation by PBMC, while optimal MAM-induced proliferation is significantly lower in magnitude . In all 12 subjects tested, mitogenic concentrations of MAM reproducibly stimulate unselected PBMC to secrete polyclonal IgM and IgG . In contrast, the S . aureus-derived SA induce Ig production only in cultures containing isolated B cell populations and either very low numbers of untreated autologous T cells, larger numbers of X-irradiated autologous T cells, or very low concentrations of the SA . No difference in the activation of helper (CD4) versus suppressor/cytotoxic (CD8) T cells by MAM and the S . aureus-derived SA was noted . Taken together, these data suggest that MAM's capacity to induce B cell differentiation correlates with its induction of a relatively weak proliferative response by unselected human T cells . MAM-like SA, when encountered in vivo, may result in a significant perturbation of the human immune system and potentially contribute to clinical syndromes characterized by immunostimulation and hypergammaglobulinemia. J Urol (Paris), 1992, 98(4), 228 - 31 {Kidney carbuncle: diagnostic, bacteriological and therapeutic considerations . Apropos of 11 cases}; Jemni L et al.; Renal carbuncle is a cortical lesion following bacteremia, occurring in patient without any urologic known problems . The analysis of 11 consecutive renal carbuncles showed that one should consider the diagnosis of renal carbuncle in young patients with flank pain, fever, and absence of significant leucocyturia . Our study confirms that renal carbuncle is always caused by staphylococcus aureus and that treatment is based on appropriate antibiotherapy . Isolation of the bacteria was difficult unless ponction of carbuncle under ultrasound control was performed . The usefulness of ultrasonography for the diagnosis of renal carbuncle and for its distinction from other suppurative renal lesions is emphasized. Acta Haematol Pol, 1992, 23(4), 253 - 8 {Evaluation of phagocytic activity of peripheral blood neutrophils in patients after kidney transplantation treated with cyclosporin and prednisone or with azathioprine with prednisone}; Hrycek A et al.; In 32 renal transplant patients neutrophil phagocytic activity was evaluated using living bacterial cells of Staphylococcus aureus Oxford 209 P strain . The total group of patients was divided into two subgroups according to the kind of the applied immunosuppressive treatment 1) cyclosporin--12 patients treated with cyclosporin A along with prednisone and 2) azathioprine--20 patients treated with azathioprine along with prednisone . The index of phagocytizing cells i.e . the percentage of neutrophils phagocytizing the bacterial cells was assessed in the investigation of phagocytosis . In the renal transplant patients as compared with the control group, statistically significant increase in the phagocytic of peripheral blood neutrophils in relation to the used bacterial cells of Staphylococcus aureus Oxford 209 P strain was found . Statistically significant differences in the investigated range were also present between the compared subgroups . In the cyclosporin subgroup statistically significant increase was revealed in the percentage of phagocytizing neutrophils. J Hyg Epidemiol Microbiol Immunol, 1992, 36(3), 327 - 36 In vitro stimulation of human lymphocytes by alpha, beta and delta toxins and toxoids of Staphylococcus aureus; Prokesova L et al.; Alpha, beta and delta toxins of Staphylococcus aureus stimulate human peripheral blood lymphocytes to blastic transformation and formation of IgM, IgG and IgA . The toxins are efficient at concentrations that are not toxic for the cells in culture . A dose of a toxin suitable for stimulation is 100 ng/ml but a stimulation can be observed also at 10 ng/ml, in the case of Ig formation even at a concentration of 1 ng/ml . Toxoids are approximately as effective to elicit blastic transformation as the toxins themselves, their efficiency to stimulate Ig formation being somewhat lower but significant . Alpha and delta toxins and toxoids at the appropriate concentration appear to act as medium-strength polyclonal activators of lymphocytes . Beta toxin and its toxoid are weak polyclonal activators. Microbiol Immunol, 1992, 36(12), 1233 - 7 An improved method for the serotyping of free coagulase from Staphylococcus aureus; Tajima Y et al.; The serotyping of free coagulase, one of the most reliable ways to identify strains of Staphylococcus aureus, and widely employed in Japan, has been improved by adding magnetite sand to the reaction mixture . Culture medium supernatant and a type-specific antibody are mixed in a well of a microtiter plate, and plasma-enriched bovine fibrinogen is treated with magnetite sand . The use of tranexamic acid and gum arabic in the reaction mixture also increases the sensitivity of the reaction . Finally, the plate is placed on a magnetic stirrer . If the type of the coagulase corresponds to that of the antibody, no clot formation will occur, and this is easily confirmed by the movement of the sand . Although the amount of reaction mixture required is much less than that for the conventional tube method, our new method is able to detect slight increases in viscosity of the reaction mixture due to fibrin formation even before complete clotting occurs, thus providing very high sensitivity . Clot formation can also be judged by observing a turbid mass of fibrin in the well (Hwang's method), but this approach is a little slower than our method involving immobilization of magnetite sand. Exp Clin Immunogenet, 1992, 9(2), 109 - 16 Failure of c-myc gene expression in B cells of some patients with common variable immunodeficiencies; Kondo N et al.; Many reports have shown that expression of the c-myc protooncogene represents an early event of lymphocyte activation . Calcium influx and activation of protein kinase C synergistically bypass the early signal transduction of lymphocyte activation . In this study, the c-myc message of B cells or B cell lines stimulated by 12-o-tetradecanoylphorbol-13-acetate (TPA), A23187, Staphylococcus aureus Cowan I (SAC), or anti-mu was not expressed or was poorly expressed in common variable immunodeficiency (CVID) patients whose B cells did not differentiate or only poorly differentiated to SAC plus recombinant interleukin 2, whereas the c-myc message of 1 CVID patient's B cells that differentiated well in IgM secretion to SAC plus recombinant interleukin 2 was well expressed when stimulated by TPA, A23187, SAC, or anti-mu . These results suggest that an abnormality exists in the early signal transduction process on some CVID patients' B cells and that it may be in the bypass by calcium influx and direct activation of protein kinase C. Cytobios, 1992, 71(284), 29 - 36 Regulation of Staphylococcus protease using complement, interferon and immunoglobulin as substrates; Kuo WN et al.; The effects of various agents on the cleavage of serum albumin, interferon, immunoglobulin and complement component C1q by the extracellular protease from Staphylococcus aureus were analysed by SDS-polyacrylamide gel electrophoresis . Arachidonic acid moderately stimulated the proteolysis of serum albumin, interferon and complement component . Phosphatidic acid effectively enhanced the proteolysis of serum albumin and IgG, whereas it inhibited the cleavage of IgM . The proteolysis of IgG was appreciably enhanced by sphingosine . In contrast, phosphatidyl choline and phosphatidyl glycerol were shown to have an inhibitory effect on the proteolysis of IgG and IgM . Phosphatidyl serine, phosphatidyl inositol and phosphatidyl ethanolamine also inhibited the proteolysis of IgG . The failure of any of these agents to exert a persistent effect on the cleavage of all substrates, revealed the complexity of the interactions among the agent, the substrate and the protease. J Biomater Sci Polym Ed, 1992, 4(2), 89 - 97 Inhibition by heparin and derivatized dextrans of Staphylococcus aureus adhesion to fibronectin-coated biomaterials; Vaudaux P et al.; Recent data on cardiovascular device-centered infections suggest that some plasma and extracellular matrix proteins contribute to bacterial adhesion and colonization on biomaterials . We previously developed an in vitro assay to study the Staphylococcus aureus adhesion-promoting effect of surface-adsorbed fibronectin on flat PMMA coverslips coated with a monolayer amount of fibronectin . We screened the potential anti-adhesive properties of a group of substituted dextrans, previously shown to exhibit potent anticoagulant and anticomplementary activities . In comparison to unsubstituted dextran which showed no significant (< 20%) adhesion inhibition at 1 mg/ml, dextrans increasingly substituted with carboxylic and benzylamide groups (CMBD) exhibited increasing anti-adhesive activities . Three CMBD derivatives showing an increasing proportion (5-14%) of benzylamide groups showed inhibition of bacterial adhesion increasing from 33 to 51% at 1 mg/ml . Another category of substituted dextrans having a variable proportion (2-26%) of sulfonated benzylamide groups (CMBDS) produced active inhibition of S . aureus adhesion . In comparison to these heparin-like dextran derivatives, native heparin produced inhibition values of S . aureus adhesion which were intermediate between those of CMBD and CMBDS compounds . Furthermore, the anti-adhesive activity was still expressed when substituted dextrans were preincubated with fibronectin-coated PMMA but washed away at the time when radiolabeled bacteria were added to the adhesion assay . This indicates that the anti-adhesive effects of CMBDS could be exerted at the level of the S . aureus binding site of fibronectin . In conclusion, S . aureus adhesion on fibronectin-coated biomaterials can be efficiently blocked in vitro by soluble compounds such as dextran derivatives. J Cardiovasc Pharmacol, 1992, 20 Suppl 12, S145 - 7 Lipoteichoic acid: a new inducer of nitric oxide synthase; Lonchampt MO et al.; Inducible nitric oxide (NO) synthase in vascular smooth muscle cells (SMCs) appears to play a major role for the diminished responsiveness to vasoconstrictors observed in endotoxemia . However, cardiovascular dysfunctions associated with septic shock are also observed in the absence of endotoxin (LPS) . Similar hemodynamic changes are produced either by a gram-negative bacteria (Escherichia coli) or by a gram-positive bacteria (Staphylococcus aureus), a microorganism without LPS, suggesting a common pathway leading to cardiovascular abnormalities . In the present study, we describe the induction of NO synthase in vascular SMCs by lipoteichoic acid (LTA), a component of the membrane of gram-positive bacteria . In cultured vascular SMCs, a 24-h incubation with LTA produced an increase in intracellular cyclic GMP . This effect was inhibited by methylene blue (MB), an inhibitor of guanylate cyclase . Incubation with a specific inhibitor of L-arginine, i.e., NG-nitro-L-arginine methyl ester (L-NAME), or depletion of L-arginine attenuated the LTA-induced cGMP production . A 5-h incubation of endothelium-free rings of rat aorta in the presence of LTA induced a loss of tonicity to the contractile response of phenylephrine . The contractions were restored by MB and by L-NAME . The effect of L-NAME was reversed by L-arginine . These results show that LTA, like LPS, expresses NO synthase in vascular SMCs. Ann Trop Paediatr, 1992, 12(3), 265 - 71 Paediatric empyema thoracis in Zaria, Nigeria; Hassan I et al.; From 1977 to 1986 at the Ahmadu Bello University Hospital in Zaria, the diagnosis of empyema thoracis was made in 150 children representing 0.35% of all paediatric admissions . Empyema was a complication of pneumonia in 98 of 100 cases reviewed, of whom 70 were 3 years old or younger . There was no sex or side preference . Fifty-nine patients were severely ill on admission, with a mean symptom duration of 21 days . In many of them, anaemia, protein-energy malnutrition, measles and gastro-enteritis were present as associated illnesses . Staphylococcus aureus was the most frequent bacterial isolate (39%), of which 83% of the cultures were sensitive to cloxacillin . Adequate rehydration, systemic antibiotics and tube thoracostomy drainage performed in 80 patients were more successful than repeated needle aspiration . Fourteen of the children died, six absconded and 80 were discharged home. J Mol Biol, 1991 Dec 20, 222(4), 857 - 9 Crystallization and preliminary X-ray analysis of the lactose-specific phosphocarrier protein IIAlac of the phosphoenolpyruvate: sugar phosphotransferase system from Staphylococcus aureus; Celikel R et al.; The IIA constituent of the lactose permease from Staphylococcus aureus has been crystallized in two different forms . Crystals of form I have been grown from polyethylene glycol 4000 with beta-octyl glucoside . They diffract to 3.0 A resolution and belong to space group C2 with unit cell dimensions a = 141.7 A, b = 130.7 A, c = 96.5 A and beta = 96.2 degrees . Form II crystals have been obtained from a solution containing polyethylene glycol 400, ammonium sulfate and manganese chloride . They diffract to at least 2.8 A resolution and belong to space group P2(1)2(1)2(1) with unit cell dimensions a = 89.9 A, b = 101.5 A and c = 90.9 A. Am J Med, 1991 Dec 30, 91(6A), 27S - 30S Mechanisms and frequency of resistance to temafloxacin; Appelbaum PC; The bactericidal activity of temafloxacin and other fluoroquinolones is attributed to inhibition of bacterial DNA gyrase (topoisomerase II), an enzyme that regulates supercoiling and uncoiling of DNA . Because of this unique bactericidal mechanism of action, resistance to fluoroquinolones is limited to spontaneous mutations . In vitro studies were conducted to determine the frequency at which spontaneous resistance to temafloxacin occurs and to determine whether cross-resistance to other antimicrobial agents develops . In 13 strains of bacteria, the frequency of spontaneous resistance mutation to concentrations of temafloxacin at four and eight times the minimal inhibitory concentration (MIC) ranged from less than 1 x 10(-10) to 1.4 x 10(-7) . Temafloxacin demonstrated a lower frequency of resistance to both methicillin-resistant and methicillin-sensitive Staphylococcus aureus compared with ciprofloxacin and ofloxacin . Cross resistance with other fluoroquinolones was observed. Am J Med, 1991 Dec 30, 91(6A), 15S - 18S In vitro activity of temafloxacin against gram-positive cocci including methicillin-resistant Staphylococcus aureus; Fuchs PC; Published reports on the in vitro activity of temafloxacin indicate its activity against gram-positive cocci is equal to or slightly greater than that of ciprofloxacin, and generally twofold greater than that of ofloxacin . The few ciprofloxacin-resistant Staphylococcus aureus strains reported were also resistant to temafloxacin . Because temafloxacin achieves higher plasma and tissue levels than ciprofloxacin, greater differences between minimal inhibitory concentrations and tissue levels occur with temafloxacin than with ciprofloxacin, which may provide temafloxacin with greater therapeutic efficiency against gram-positive coccal infections. J Biol Chem, 1991 Dec 25, 266(36), 24540 - 8 Determinants of the phagosomal pH in macrophages . In situ assessment of vacuolar H(+)-ATPase activity, counterion conductance, and H+ "leak"; Lukacs GL et al.; We studied the factors that determine the intraphagosomal pH (pHp) in elicited murine peritoneal macrophages . pHp was measured in situ by recording the fluorescence of covalently fluoresceinated Staphylococcus aureus ingested by the macrophages . Following spontaneous acidification of the phagosomes, passive (leak) H+ permeability was determined measuring the rate of change of pHp upon complete inhibition of the H+ pump with bafilomycin A1 . A significant, but comparatively low passive H+ permeability was detected . The existence of a passive H+ leak implies that continuous energy expenditure is required for the maintenance of an acidic pHp . In combination with ionophores, bafilomycin was also used to estimate the counterion permeability . The counterion conductance was found to be severalfold higher than the H+ leak . Ion substitution experiments in electropermeabilized cells and the inhibitory effects of quinine and 5-nitro-2-(3-phenylpropylamino)benzoic acid suggest that both monovalent anions and cations permeate the phagosomal membrane . The activity of the H+ pump was measured at various pHp levels . In the steady state, the rate of H+ pumping was considerably lower than counterion permeation . These findings suggest that the phagosomal membrane potential is insignificant . Consistent with this notion, increasing phagosomal conductance with ionophores failed to accelerate the rate of H+ pumping . Thus, the transmembrane delta pH is the predominant component of the proton-motive force across the phagosomal membrane in the steady state . The rate of H+ pumping was found to decrease steeply as the phagosomal lumen became acidified . Therefore, the pH sensitivity of the H+ pump, which possibly reflects a kinetic or allosteric effect, is the primary determinant of pHp. Biochemistry, 1991 Dec 24, 30(51), 11742 - 50 Primary structure of the catalytic subunit of calf thymus DNA polymerase delta: sequence similarities with other DNA polymerases; Zhang J et al.; The 125- and 48-kDa subunits of bovine DNA polymerase delta have been isolated by SDS-polyacrylamide gel electrophoresis and demonstrated to be unrelated by partial peptide mapping with N-chlorosuccinimide . A 116-kDa polypeptide, usually present in DNA polymerase delta preparations, was shown to be a degraded form of the 125-kDa catalytic subunit . Amino acid sequence data from Staphylococcus aureus V8 protease, cyanogen bromide, and trypsin digestion of the 125- and 116-kDa polypeptides were used to design primers for the polymerase chain reaction to determine the nucleotide sequence of a full-length cDNA encoding the catalytic subunit of bovine DNA polymerase delta . The predicted polypeptide is 1106 amino acids in length with a calculated molecular weight of 123,707 . This is in agreement with the molecular weight of 125,000 estimated from SDS-polyacrylamide gel electrophoresis . Comparison of the deduced amino acid sequence of the catalytic subunit of bovine DNA polymerase delta with that of its counterpart from Saccharomyces cerevisiae showed that the