J Trauma, 1985 Oct, 25(10), 1004 - 6 Toxic shock syndrome in a scald burn victim; Farmer BA et al.; A child with 12% total body surface area superficial and partial-thickness burns was admitted to the Oregon Burn Center . Within 48 hours of admission, signs and symptoms of toxic shock syndrome (TSS) were present including high spiking fevers, vomiting, diarrhea, hypotension, conjunctival hyperemia, and a diffuse macular erythroderma . Cultures of skin pustules and burn wounds grew Staphylococcus aureus . This strain has been shown to produce staphylococcal enterotoxin B (SEB) . This case appears to be the first reported of toxic shock syndrome in a burn victim caused by staphylococcal enterotoxin B. J Med Microbiol, 1985 Oct, 20(2), 275 - 8 The expression of capsule in serum-soft agar by Staphylococcus aureus isolated from human clinical sources; Opdebeeck JP et al.; Staphylococcus aureus isolates from human clinical sources were incubated for various times in modified 110 medium and tested for production of capsule by the serum-soft agar technique . Ten (5.7%) of 175 isolates were encapsulated after incubation for 24 h . A more detailed examination of 77 isolates showed that incubation period affected the production of capsule . After 2 h, 31% were encapsulated, but after 6 h and 24 h this decreased to 17% and 4% respectively . Rapid passage in vitro induced the expression of capsule in four of 50 unencapsulated strains . Only three of 20 encapsulated strains could be typed with standard antisera. J Med Microbiol, 1985 Oct, 20(2), 249 - 53 Protein-mediated adhesion of Staphylococcus aureus to silicone implant polymer; Barrett SP; Investigation was made of the role of protein A and clumping factor in the adhesion of Staphylococcus aureus to the silicone polymer used for manufacture of cerebrospinal fluid shunting systems . The two proteins were judged to contribute non-specifically to adhesion . S . aureus was also shown to be capable of hydrophobic binding, but this was found to be distinct from the demonstrated protein-mediated adhesion. Infect Immun, 1985 Oct, 50(1), 304 - 9 Expression of the cloned toxic shock syndrome toxin 1 gene (tst) in vivo with a rabbit uterine model; de Azavedo JC et al.; Toxic shock syndrome (TSS) toxin 1 (TSST1) is produced by strains of Staphylococcus aureus associated with TSS . Purified TSST1 induces in rabbits a shock-like illness with many features similar to TSS in humans . These symptoms were also induced by TSST1-producing bacteria in diffusion chambers implanted in the rabbit uterus . Naturally occurring TSST1+ strains and a TSST1- strain harboring a pE194-derived plasmid carrying the cloned TSST1 determinant tst gave the same symptoms . TSST1- strains and a TSST1- strain carrying a pE194-tst plasmid with a deletion of the tst gene had no effect in rabbits . The results with the plasmid-carrying TSST1+ and TSST1- strains, which were isogenic apart from tst, show that the toxin is responsible for the illness in rabbits and suggest that it is a major factor in the pathogenesis of TSS. J Clin Microbiol, 1985 Oct, 22(4), 501 - 4 Evaluation of mannitol salt agar with oxacillin as a screening medium for methicillin-resistant Staphylococcus aureus; Lally RT et al.; We evaluated the use of mannitol salt agar with oxacillin for use as a primary screening medium for the simultaneous detection and identification of methicillin-resistant Staphylococcus aureus in clinical surveillance specimens . Oxacillin agar dilution susceptibility tests with mannitol salt agar and Mueller-Hinton agar were performed in parallel with disk-agar diffusion testing on 95 oxacillin-susceptible and 105 oxacillin-resistant S . aureus stock isolates . MICs were found to be comparable, showing distinct separation of susceptible and resistant isolates into two groups with MICs of less than or equal to 2 and greater than or equal to 32 micrograms/ml, respectively . In accord with these findings, 4 micrograms of oxacillin per ml was selected for use in the screening medium . For performance evaluation, mannitol salt agar with 4 micrograms of oxacillin per ml was compared with mannitol salt agar without oxacillin by performing parallel screening tests on 153 clinical surveillance specimens . For detection of methicillin-resistant S . aureus, mannitol salt agar with 4 micrograms of oxacillin per ml was as sensitive as mannitol salt agar without oxacillin and required significantly fewer confirmatory tests . For primary identification of methicillin-resistant S . aureus, mannitol salt agar with 4 micrograms of oxacillin per ml was 6.4% false-positive and 1.1% false-negative, with a 93.6% positive predictive value . These findings indicate that mannitol salt agar with 4 micrograms of oxacillin per ml can be used as a reliable and cost-effective screening medium for the simultaneous detection and identification of methicillin-resistant S . aureus in clinical surveillance specimens. Clin Exp Immunol, 1985 Oct, 62(1), 112 - 20 OKT4+ and OKT8+ T lymphocytes produce soluble factors that can modulate growth and differentiation of human B cells; Sakane T et al.; The human T cell subset(s) responsible for the production of soluble factors that can modulate B cell growth and B cell differentiation was investigated in the present study . For this purpose, highly purified OKT4+ and OKT8+ lymphocytes were stimulated with phytohaemagglutinin and phorbol myristate acetate . Subsequently, culture supernatants were analysed for B cell growth factor (BCGF) activity and B cell differentiation factor (BCDF) activity in the following systems: 1) maintenance of a proliferative state of Staphylococcus aureus of strain Cowan I (SAC)-stimulated B cells and 2) induction of plaque-forming cell responses of SAC-stimulated B cells . Mitogenic stimulation led to production of equivalent amounts of either BCGF activity or BCDF activity from both of the OKT4+ and OKT8+ subsets . These findings may provide a basis for further studies of the molecular mechanisms as well as cellular interactions involved in human B cell activation, proliferation and differentiation. Antimicrob Agents Chemother, 1985 Oct, 28(4), 467 - 72 Double-blind, placebo-controlled study of oxacillin combined with rifampin in the treatment of staphylococcal infections; Van der Auwera P et al.; A total of 101 patients with proven Staphylococcus aureus infection were included in a double-blind, placebo-controlled study; this study compared oxacillin (12 g/day, intravenously) or vancomycin (2 g/day, intravenously) plus rifampin (1,200 mg/day, orally) with oxacillin or vancomycin plus placebo . We evaluated 65 patients . Of the patients tested, 33 received oxacillin plus rifampin (13 bacteremias), and 32 received oxacillin plus placebo (16 bacteremias) . Clinical cure was achieved in 61% of the patients treated with oxacillin plus rifampin and in 56% of the patients treated with oxacillin plus placebo . Improvement was noted in 27 and 25%, respectively, and failure occurred in 9 and 18%, respectively . These differences were not statistically significant . Bacteriological failure occurred in 3 and 28%, respectively (P less than 0.05) . None of the failures within the rifampin-treated group was associated with the emergence of a rifampin-resistant mutant . The rates of superinfection were similar in both groups . The geometric means of the serum bactericidal activity after 1, 6, and 11 h were, respectively, 22, 17, and 9 after treatment with oxacillin plus rifampin and 25, 3.4, and 2.3 after treatment with oxacillin plus placebo . It was suggested that the addition of rifampin to oxacillin or vancomycin might only be beneficial to severely ill patients. Eur J Clin Microbiol, 1985 Oct, 4(5), 478 - 82 Comparison of a biphasic medium plus routine early subculture with a slide blood culture system; Degaute C et al.; The accuracy of a biphasic medium plus routine early subculture on chocolate agar was compared with that of a slide blood culture system for detecting facultatively anaerobic bacteria in blood cultures . Over a three month period 2,745 pairs of blood cultures were examined . Bacteremia was diagnosed in 91 patients . Twelve cases would have been missed by using the slide blood culture system alone, in contrast to only three by the biphasic medium alone . The two techniques detected the same amount of contaminant bacteria . Although the slide blood culture system required less time in general, the biphasic medium proved more accurate for isolating Staphylococcus aureus (p = 0.005) and detected gram-positive bacteria more rapidly (p = 0.002). J Bone Joint Surg Am, 1985 Oct, 67(8), 1236 - 44 The influence of skeletal implants on incidence of infection . Experiments in a canine model; Petty W et al.; We have performed experiments in 187 dogs in order to evaluate the effect of commonly used implant materials on rate of infection . We opened the femoral canal with a hand drill and awl, instilled a suspension of bacteria, and then inserted one of the implants . The implants--stainless-steel and cobalt-chromium alloys, high-density polyethylene, prepolymerized polymethylmethacrylate, and polymethylmethacrylate polymerized in vivo--were compared with no implant (control) . The effect of the different implants on the incidence of infection with Staphylococcus epidermidis, Staphylococcus aureus, and Escherichia coli was compared by determining the number of bacteria required to produce infection in 50 per cent of the femora . All of the implants were significantly more likely than the controls to be associated with infection with Staphylococcus aureus . Polymethylmethacrylate polymerized in vivo was found to be significantly more likely than all other implants to be associated with infection with Escherichia coli and Staphylococcus epidermidis . In addition to evaluating all specimens bacteriologically, we carried out a histological evaluation, and found that infection was highly correlated with an increased inflammatory response for all three bacteria . However, even with this highly statistically significant correlation, the correlation was not absolute; when only limited portions of randomly selected specimens of tissue were examined, the correlation was reduced. Am Surg, 1985 Oct, 51(10), 577 - 9 Infectivity of vascular sutures; Scher KS et al.; Bacterial adherence to vascular sutures was evaluated in vitro using radioactively labeled Staphylococcus aureus . The following suture materials were tested: polypropylene, silicone-treated braided polyester, and Teflon-treated braided polyester . Significantly fewer bacteria adhered to the monofilament polypropylene than either of the braided polyester sutures . There was no significant difference between silicone-treated and Teflon-treated polyester from the standpoint of bacterial adherence . Vascular sutures were evaluated in vivo using a mouse wound model . Sutures were tested with and without knots . When tested without knots, fewer bacteria were recovered from wounds containing polypropylene suture compared to either of the braided materials, although this apparent advantage did not prove to be statistically significant . When studied with knots, the differences among types of suture were much less marked and, again, not significant . The purported contribution of the monofilament structure of a suture to its infection resistance may have been overstated. Immunol Invest, 1985 Oct, 14(5), 421 - 6 Induction of sIg-negative B lymphocytes for differentiation by PWM; Zhu LP et al.; It has been reported in the literature that PWM-responsive B lymphocytes are sIgM+, sIgD-, lack receptors for mouse erythrocytes, and are larger than PWM-negative lymphocytes . This paper describes the observation of B cells among the thymocytes from a patient with myasthenia gravis . In the thymocytes, actually no sIg+ cell was found, and Staphylococcus aureus Cowen strain I could not induce the mononuclear cells in the thymus to differentiate . It was surprising that 6.9 per cent of the cells were BA-1 positive cells . At the same time, in the presence of PWM, 9680 +/- 555 cells out of 1 X 10(-6) thymocytes could be induced to differentiate to immunoglobulin-secreting cells (ISC) in culture . This result suggests that PWM might be able to induce sIg- B lymphocytes to differentiate as well. Blood, 1985 Oct, 66(4), 848 - 58 The expression of the Hodgkin's disease associated antigen Ki-1 in reactive and neoplastic lymphoid tissue: evidence that Reed-Sternberg cells and histiocytic malignancies are derived from activated lymphoid cells; Stein H et al.; Ki-1 is a monoclonal antibody (raised against a Hodgkin's disease-derived cell line) that, in biopsy tissue affected by Hodgkin's disease, reacts selectively with Reed-Sternberg cells . The expression of Ki-1 antigen has been analyzed by immunocytochemical techniques in a wide range of human tissue and cell samples, including fetal tissue, malignant lymphomas (290 cases), and mitogen- and virus-transformed peripheral blood lymphocytes . The antigen was detectable on a variable proportion of cells in all cases of lymphomatoid papulosis and angio-immunoblastic lymphadenopathy and in 28% of the cases of peripheral T cell lymphomas (including lympho-epithelioid lymphomas) . It was also expressed (more strongly) on tumor cells in 45 cases of diffuse large-cell lymphoma, most of which had originally been diagnosed as malignant histiocytosis or anaplastic carcinoma, because of their bizarre morphology . However, all of these cases lacked macrophage and epithelial antigens . Thirty-five cases expressed T cell-related antigens (associated in nine cases with the coexpression of B cell-related antigens), seven bore B cell-related antigens alone, and three were devoid of T and B cell markers . DNA hybridization with a JH specific probe showed a germline configuration in 11 cases of T cell phenotype, in two cases lacking T and B cell antigens, and in one case of mixed T/B phenotype, while rearrangement was found in two cases of clear B cell type and in one mixed T/B case . Expression of the Ki-1 antigen could be induced, together with interleukin 2 (IL 2) receptor, on normal lymphoid cells of both T and B cell type by exposure to phytohemagglutinin, human T leukemia viruses, Epstein-Barr virus, or Staphylococcus aureus . The results obtained indicate that Ki-1 antigen is an inducible lymphoid-associated molecule that identifies a group of hitherto poorly characterized normal and neoplastic large lymphoid cells . Tumors comprised solely of these cells show both morphological and immunological similarities to the neoplastic cells in Hodgkin's disease . This suggests that both disorders represent the neoplastic proliferation of activated lymphoid cells of either T cell or, less commonly, B cell origin . Disorders in which only a minority of cells express Ki-1 antigen (lymphomatoid papulosis, angio-immunoblastic lymphadenopathy, and certain T cell lymphomas) probably represent lesions in which only some of the abnormal cells have transformed into an "activation state." In direct support of this view is the finding that the Ki-1 expression in these lesions is accompanied by the expression of HLA-DR and IL 2 receptors. Clin Immunol Immunopathol, 1985 Oct, 37(1), 13 - 21 Staphylococcal protein A bound to Sepharose 4B is mitogenic for T cells but not B cells from rabbit tissues; Kraft SC et al.; The mitogenicity of protein A from Staphylococcus aureus bound to Sepharose 4B (SpA-S) was tested against mononuclear cells from normal rabbit spleens and Peyer's patches after using Sephadex G-10 adherence chromatography to deplete macrophages and Sephadex G-200 anti-rabbit F(ab')2 immunoabsorbent columns to obtain B-enriched lymphocytes . Macrophage-depleted unseparated lymphocytes and the B-cell-depleted (T-cell-enriched) fractions from both tissues consistently showed good mitogenic responses to SpA-S . In contrast, the B-cell-enriched populations from either tissue did not respond to SpA-S . Supplementary experiments employing glass adherence and T-cell autorosetting did not support the possibility that the more efficient immunoabsorbent method of B-cell isolation had in some way caused the unresponsiveness to SpA-S . These results indicate that SpA-S is mitogenic for rabbit tissue T cells but will not serve as a T-independent mitogen for rabbit tissue B cells. J Med Microbiol, 1985 Oct, 20(2), 139 - 45 A resistance determinant to nucleic acid-binding compounds in methicillin-resistant Staphylococcus aureus; Emslie KR et al.; Recent isolates of methicillin-resistant Staphylococcus aureus from Australia and several other countries carry a plasmid coding for high levels of resistance to propamidine isethionate and low levels of resistance to cetyltrimethyl-ammonium bromide . The reasons for the acquisition and retention of such a determinant are not known . To define the properties of this resistance determinant in more detail, the minimum inhibitory concentrations of a series of cationic agents, including ethidium bromide, were determined and a cationic-resistance profile prepared for several strains of S . aureus and their isogenic, sensitive derivatives . The compounds for which resistance is coded by this determinant all bind to nucleic acids . Hence the determinant has been designated the NAB (nucleic acid-binding compounds)-resistance determinant. Ric Clin Lab, 1985 Oct-Dec, 15(4), 323 - 9 Selective C3 deficiency due to C3 nephritic factor in an apparently healthy girl; Tedesco F et al.; Routine laboratory investigations performed on the serum of an 8-year-old girl examined because of a moderate degree of iron-deficiency anemia showed a markedly reduced C3 level . More detailed complement studies revealed a selective C3 deficiency, as indicated by the almost undetectable C3 concentration tested by both hemolytic and immunochemical assays and by the normal or slightly reduced levels of all the other complement components . The hemolytic activity of the serum was restored by the addition of partially purified C3 component . The isolated C3 deficiency could be attributed to the presence of a C3-cleaving activity in the serum of the propositus . This activity was identified as C3 nephritic factor (C3NeF) since it was heat-stable, was absorbed by Cowan I strain of Staphylococcus aureus and was-eluted in the IgG fraction after DEAE-chromatography of the serum . The levels of H and I factors of the alternative pathway in the serum of the propositus and of C3 in the serum samples of her parents and two siblings were found to be within the normal range . The previous clinical history of the girl and the follow-up for a period of approximately 5 years showed that she was apparently healthy and did not reveal clinical and/or laboratory evidence of glomerulonephritis, lipodystrophy or repeated bacterial infections usually associated with the presence of C3NeF in the serum. Immunology, 1985 Oct, 56(2), 329 - 35 Autocrine models of B-lymphocyte growth . I . Role of cell contact and soluble factors in T-independent B-cell responses; Gordon J et al.; The requirements for triggering human B cells to DNA synthesis by T-independent polyclonal activators were examined . Optimal S phase entry of purified resting B cells infected with Epstein-Barr virus (EBV) or confronted with killed particles of Staphylococcus aureus Cowan Strain I (SAC) required a high density of cells in culture . Experiments varying culture vessel geometry and culture volumes revealed that the initial limiting quantity was a soluble activity generated in the B-cell cultures . A parallel observation was noted in the requirements for the sustained growth of EBV-transformed lymphoblasts . Autostimulatory soluble factors harvested from such cultures were able to augment DNA synthesis in low density cultures of resting cells triggered by EBV or SAC . Below a critical cell number, however, soluble factors by themselves, were not sufficient either for supporting primary B-cell responses or for maintaining the proliferation of transformed lymphoblasts . By employing conditions which encouraged cell contact it was found that a second, non-harvestable factor requiring cell proximity for its action was also necessary to promote B-cell growth . The implications of these findings for autocrine and paracrine models of B-cell activation are discussed. J Med Microbiol, 1985 Oct, 20(2), 157 - 67 Susceptibility to antimicrobial agents and analysis of plasmids in gentamicin- and methicillin-resistant Staphylococcus aureus from Dublin hospitals; Coleman DC et al.; Methicillin- and gentamicin-resistant Staphylococcus aureus (MGRSA) strains isolated from Dublin Hospitals were classified into two groups (phenotypes) . Phenotype-I strains expressed high level resistance to gentamicin and were susceptible to fusidic acid; strains resistant to tetracycline harboured a 3 X 10(6)-mol . wt plasmid . Strains in phenotype II usually expressed low level resistance to gentamicin, were resistant to fusidic acid and often harboured a (22-24) X 10(6)-mol . wt plasmid that specified resistance to ethidium bromide, tetracycline, kanamycin, neomycin and trimethoprim, or to combinations of these markers . A few phenotype-II strains expressed higher levels of resistance to gentamicin and other aminoglycosides . All MGRSA strains carried a 21 X 10(6)-mol . wt plasmid conferring resistance to penicillin, ethidium bromide, cadmium and mercury . Gentamicin resistance was invariably chromosomal and all strains carried chromosomal resistance to methicillin, erythromycin, streptomycin and spectinomycin . Several methicillin-resistant S . aureus (MRSA) strains isolated before the emergence of gentamicin resistance harboured a 21 X 10(6)-mol . wt penicillinase plasmid with the same restriction endonuclease profile as that from some MGRSA strains . Some MRSA strains carried other plasmids related to those found in MGRSA strains. J Gen Virol, 1985 Oct, 66 ( Pt 10), 2279 - 83 A hypothesis accounting for the effect of the host cell on neutralization-resistant virus; Kjellen L; Evidence is presented showing that a monkey anti-enterovirus 71 immune serum contains several antibody populations which differ in their mode of function . One population reduces infectivity, although inefficiently, by interactions at exposed antigenic sites and can be detected by measuring residual virus infectivity after mixtures of virus and antibody have been allowed to interact . Another antibody population, which is unaffected by the immunosorbent Staphylococcus aureus (Cowan I strain), appears to attach to its antigenic site(s) only after interactions between enterovirus 71 and host cells have already begun . In view of the transience of (presumed) conformational changes in the invading viruses, demonstration of this type of antibody activity requires a particular host cell system . This second type of antibody neutralization could be detected on RD cells but not on green monkey kidney cells. J Virol, 1985 Oct, 56(1), 298 - 302 Proteins associated with human parainfluenza virus type 3; Jambou RC et al.; The polypeptides associated with human parainfluenza virus type 3 were identified . Five proteins were present in detergent- and salt-resistant viral cores . Of these, three proteins designated NP0, NP1, and NP2 of 68,000, 58,000, and 52,000 daltons, respectively, were stably associated with 50S RNA in CsCl gradient-purified nucleocapsids . The amounts of NP1 and NP2 were variable, and these proteins were shown to be structurally related to the major nucleocapsid protein (NP0) by partial Staphylococcus aureus V8 protease mapping . The other core proteins included a 240K protein designated L (candidate for the viral polymerase) and an 84K protein designated as the phosphoprotein (P) on the basis of a predominant incorporation of Pi . The viral envelope had four prominent proteins (72, 53, 40, and 12K) under reducing conditions of electrophoresis . The 72 and 53K proteins were specifically labeled with {3H}glucosamine and {3H}mannose . When sulfhydryl reagents were removed, a new 62K protein was visualized in place of the 72, 53, and 12K proteins . The 53 and 12K proteins were interpreted to be the two subunits (F1 and F2) of the fusion protein, and the 72K protein was designated as the HN (hemagglutinin-neuraminidase) glycoprotein . The unglycosylated 40K protein represented the viral matrix protein (M) . Immunoprecipitation of infected cell lysates with rabbit hyperimmune antiserum against purified virus confirmed the viral origin of these polypeptides. Zh Mikrobiol Epidemiol Immunobiol, 1985 Oct, (10), 43 - 5 {Incidence of staphylococcal carriage in pregnant women}; Kazanskaia GM et al.; In 1,494 pregnant women (on weeks 32-36) staphylococcal carriership was studied . One-third of the women under investigation were found to be carriers, their bacterial discharge increasing in spring and summer . The isolated strains proved to be polyresistant to antibiotics and belonged to epidemic strains . About a half of Staphylococcus aureus strains could not be identified with the use of the international set of phages. J Med Microbiol, 1985 Oct, 20(2), 147 - 55 Lysogenicity of methicillin-resistant strains of Staphylococcus aureus; Beard-Pegler MA et al.; The lysogenic status of 23 strains of methicillin-resistant Staphylococcus aureus, isolated at the Royal Prince Alfred Hospital, Sydney, since 1980, was studied . Twenty strains, belonging to the four predominant phage types isolated in this hospital, carried the same lysogenic phage which we have designated C . Three other phages were isolated from five strains belonging to phage type 84/85/90 . The presence of phage C had little effect on the phage-typing pattern of the strains . Similarly, lysogenization with the other three phages did not result in a significant change in phage-typing patterns . However, when strain 1489, isolated in 1969, was lysogenized with these three phages, there was a change in phage-typing pattern . Lysogenization of this strain with phage 47T resulted in a marked loss of sensitivity to both group-I and group-III phages . The lysogenic status of these methicillin-resistant strains of S . aureus was compared with that of strains isolated between 1967 and 1970 . There was no evidence that the strains isolated recently were either related to, or derived from, the earlier ones. J Bacteriol, 1985 Oct, 164(1), 397 - 400 Bacteriophages in L form of Staphylococcus aureus; Schmid EN; Lysogenicity and phage typability of Staphylococcus aureus L-form cells are described . Spontaneously produced phages were found in thin sections of S . aureus L colonies . The dimensions of the tail and head resemble those of the morphological group BIII2 (T . Krzywy, I . Durlakowa, A . Kucharewcz-Krukowska, S . Krynski, and S . Slopek, Zentralbl . Bakteriol . Mikrobiol . Hyg . Abt . 1 Orig . Reihe A 250:287-295, 1981) . Phages 3A and 3C of the international typing set lysed both the bacillary and L form of S . aureus. Acta Pathol Microbiol Immunol Scand {B}, 1985 Oct, 93(5), 359 - 63 Hydrophobized wound dressing in the treatment of experimental Staphylococcus aureus infections in the young pig; Wadstrom T et al.; Standard wounds (2 X 2 cm) produced with a dermatome, followed by a thermal injury in four-week-old piglets were inoculated with Staphylococcus aureus strains . Infection developed in each wound within a few days . Wounds were then dressed separately with cellulose dressing or the same dressing substituted with a hydrophobic ligand (stearylic acid) . Other wounds were treated with active charcoal dressing (ActisorbR) or DebrisanR . Signs of healing, including vanishing of inflammation around the wounds, were noticed already on days 3 and 4 in the wounds treated with the hydrophobized dressing, and no infection was noticed in such wounds on days 5 to 6, while all other wounds were still showing signs of infection . It is concluded that active removal of S . aureus cells in the hydrophobized dressing probably decreases the number of multiplying bacteria in wounds, causing a rapid onset of wound healing. Thorac Cardiovasc Surg, 1985 Oct, 33(5), 300 - 3 Prophylactic vancomycin versus placebo in arterial prosthetic reconstructions; Jensen LJ et al.; Vascular reconstructive surgery with the placement of prosthetic material caudal to the diaphragm is occasionally associated with postoperative wound infection . These infections often lead to amputation and can be lethal . Only a few published reports contain information on the value of prophylactic antibiotic treatment with these operations, but its use is common throughout the world . To investigate this problem, a prospective, double blind, randomized study of vancomycin versus placebo in 128 vascular graft operations caudal to the diaphragm was conducted from June, 1982 to July, 1984 . The difference in infection rate was significant (2 p = 0.0008) in favor of the vancomycin group . Fourteen wound infections (21.2%) were found in the placebo group, 3 of which (4.5%) were prosthesis infections . Among the 62 vancomycin-treated patients, one case of superficial wound infection (1.6%) and no cases of prosthesis infection were found . The most common pathogen was Staphylococcus aureus . The study has demonstrated that vancomycin, a narrow spectrum antibiotic, in an ultra-short regimen (one gram one hour before surgery and one gram 4 hours later) is an effective prophylactic agent against postoperative wound infection . Temporary and, in most cases, doses-related side effects were seen in 7.9% of the patients treated with vancomycin. J Immunol, 1985 Oct, 135(4), 2654 - 60 Fine specificity, structure, and proteolytic susceptibility of the human lymphocyte receptor for IgE; Peterson LH et al.; Properties of the Fc receptor for IgE (FC epsilon R) on cultured human B lymphoblastoid cells (RPMI 8866) were studied . Specificity for human IgE (hIgE) was demonstrated by inhibition studies with both Fc epsilon R+ intact cell and detergent-solubilized receptor preparations . No interaction of the FC epsilon R with other hIg classes or with rodent IgE was seen . In other studies, 3,3-dithiobis(sulfosuccinimidyl) propionate was used to cross-link hIgE to 125I surface-labeled 8866 cells . After detergent solubilization, the 125I receptor components were isolated by immunoprecipitation, and receptor peptides of 83 and 46 kilodalton kD were demonstrated by SDS-PAGE in the presence of reducing agents . Cross-linking performed after detergent solubilization gave identical results . Tryptic maps of the 83 and 46 kD polypeptides were identical with respect to surface-iodinated peptides; this indicates a structural homology between these components . The 83 kD component was more difficult to elute from IgE affinity columns, potentially because of an increased number of IgE binding sites per FC epsilon R molecule . Limited proteolysis studies of the purified FC epsilon R with papain and V8 protease from Staphylococcus aureus demonstrated that a 16 kD FC epsilon R fragment was rapidly produced . This component was also seen after papain treatment of intact cells, and it retained the ability to interact with anti-FC epsilon R antisera and, at least in the absence of detergent, with hIgE affinity columns . Potential relationships between the FC epsilon R and lymphokines that modulate the IgE response (IgE-binding factors) are discussed. Lancet, 1985 Sep 28, 2(8457), 705 - 8 Methicillin-resistant Staphylococcus aureus in Dublin 1971-84; Cafferkey MT et al.; Between 1971 and 1975, methicillin-resistant Staphylococcus aureus (MRSA) caused sporadic infection in eight Dublin hospitals although a case of bacteraemia was not recorded until 1976 . From then on gentamicin-resistant MRSA rapidly became endemic in Dublin hospitals . The frequency of MRSA bacteraemia reached a peak in 1979-82 but MRSA infection remains an important problem . The most effective antimicrobial agent in treatment of invasive infection was vancomycin; little drug toxicity was seen . Where appropriate, concomitant surgical treatment such as debridement and drainage was usually necessary . Infection control measures directed at eliminating carriage proved effective in reducing spread . Molecular analysis showed two distinct MRSA phenotypes with similar phage-typing patterns . Gentamicin resistance was chromosomally encoded. N Z Med J, 1985 Sep 25, 98(787), 804 - 6 Intravascular cannula-related sepsis: two years experience; Thomas MG et al.; Thirteen episodes of intravascular cannula-related sepsis were seen during a two year period . Staphylococcus aureus was the most common pathogen . Fungaemia occurred in four patients all receiving total parenteral nutrition . Culture of the cannula tip and swabs of the insertion site were useful in confirming the diagnosis . Three patients with infection due to S aureus died while receiving treatment for their infections . Care with insertion and maintenance of intravascular cannulae should reduce the frequency of this iatrogenic infection. J Biol Chem, 1985 Sep 25, 260(21), 11811 - 6 Production of antibodies directed against Escherichia coli helicase III and the molecular cloning of the helicase III gene; Matson SW et al.; Helicase III from Escherichia coli has been purified to near homogeneity using single-stranded DNA-dependent adenosine nucleoside 5'-triphosphatase activity as an assay to monitor the purification . The denatured form of this 18.5-kilodalton polypeptide, isolated on a preparative polyacrylamide gel run in the presence of sodium dodecyl sulfate, has been used as an antigen to direct the production of rabbit anti-helicase III antibodies . The antibodies obtained fail to inhibit directly either the helicase activity or the DNA-dependent adenosine nucleoside 5'-triphosphatase activity of helicase III . However, when the antigen-antibody complex is removed from solution by binding to Staphylococcus aureus cells with subsequent sedimentation, there is excellent correlation between the loss of both enzymatic activities and the loss of the helicase III polypeptide . The anti-helicase III antibodies have been used as a reagent to probe immunologically a library of E . coli DNA fragments inserted into the plasmid pBR322 for expression of the helicase III antigen . The gene encoding helicase III has been localized on a 2.0-kilobase pair PvuII-EcoRI fragment . Bacterial cells harboring a multicopy plasmid containing this fragment overproduce the helicase III antigen approximately 100-fold. Biochim Biophys Acta, 1985 Sep 20, 831(1), 1 - 7 Monoamine oxidase A and monoamine oxidase B activities are catalyzed by different proteins; Smith D et al.; Monoamine oxidases A and B (amino: oxygen oxidoreductase (deaminating) (flavin-containing), EC 1.4.3.4) have been identified in the outer membranes of rat liver mitochondria by their covalent reaction with the inhibitor, {3H}pargyline . On analysis by polyacrylamide gel electrophoresis under denaturing conditions . Monoamine oxidase A was found to migrate more slowly that monoamine oxidase B . Proteins which correspond to monoamine oxidases A and B (as identified by the electrophoretic distribution of covalently bound {3H}pargyline) were excised from the gels . Subsequent analysis showed that both monoamine oxidase A and monoamine B had been highly purified by this procedure . Electrophoretic analysis of the peptides produced by limited proteolysis with bovine trypsin, alpha-chymotrypsin, Staphylococcus aureus V8 proteinase and cyanogen bromide indicate that monoamine oxidases A and B have different amino acid sequences. J Biol Chem, 1985 Sep 15, 260(20), 11348 - 56 Monoclonal antibodies as probes for determining the microheterogeneity of the link proteins of cartilage proteoglycan; Caterson B et al.; Monoclonal antibodies were raised against Swarm rat chondrosarcoma link protein 2 . Two of the resultant hybridomas (9/30/6-A-1 and 9/30/8-A-4) were used in structural analyses of the link proteins . The 9/30/6-A-1 monoclonal antibody recognized an epitope which was only present on rat chondrosarcoma link protein 2 . This epitope was absent in rat chondrosarcoma link protein 3 obtained after trypsin or clostripain treatment of rat chondrosarcoma proteoglycan aggregate, indicating that proteolytic digestion either removed or modified the epitope . Contrasting this, the 9/30/8-A-4 monoclonal antibody recognized an epitope present in link protein(s) 1, 2, or 3 isolated from cartilage of several animal species (rat, bovine, human, and chicken) . Rat chondrosarcoma link protein 2 was digested with Staphylococcus aureus V8 protease, and the resulting peptides were fractionated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and subjected to immunolocation analyses . The 9/30/6-A-1 and 9/30/8-A-4 monoclonal antibodies recognized epitopes in two different halves of the link protein molecule . The 9/30/8-A-4 monoclonal antibody was used to identify proteolytic cleavage peptides common to the individual link proteins (1, 2, or 3) purified from cartilage proteoglycans of several animal species . Digestion of rat chondrosarcoma link protein 2 with endoglycosidase H or alpha-mannosidase increased its electrophoretic mobility to that of link protein 3 and removed or altered the determinant recognized by the 9/30/6-A-1 monoclonal antibody, indicating that a high-mannose oligosaccharide chain was part of the antigenic determinant . The 9/30/8-A-4 monoclonal recognition of epitope was unaffected by endo- or exoglycosidase treatment . Endo- and exoglycosidase treatment of bovine nasal cartilage link proteins also altered their electrophoretic mobility, indicating that high-mannose oligosaccharide structures on the various link proteins (1, 2, or 3) accounted for the microheterogeneity observed in sodium dodecyl sulfate-polyacrylamide gels. Biochemistry, 1985 Sep 10, 24(19), 5203 - 8 Comparative characterization of thyroid hormone receptors and binding proteins in rat liver nucleus, plasma membrane, and cytosol by photoaffinity labeling with L-thyroxine; Dozin B et al.; Photoaffinity labeling with underivatized thyroxine (T4) was used to identify and compare the T4 binding proteins in rat liver cytosol, nuclear extract, and purified plasma membrane . When these subcellular fractions were incubated with a tracer concentration of {125I}T4, irradiated with light above 300 nm, and individually analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the radioactivity profiles revealed the presence of T4 binding proteins of molecular masses of 70, 52, 43, 37, 30, and 26 kilodaltons (kDa) in cytosol, of 96, 56, 45, and 35 kDa in nuclear extract, and of 70, 44, and 30 kDa in plasma membrane . Competition experiments performed in the presence of a 1000-fold excess of unlabeled T4 demonstrated that these binding proteins display different hormone binding activities . The similar electrophoretic mobilities of some binding proteins present in the different subcellular fractions, i.e., the 70-, 43-45-, and 30-kDa proteins, suggested that these proteins might be identical . However, double-labeling experiments in which plasma membrane, nuclear extract, and cytosol were photolabeled with either {125I} or {131I}T4 and mixed, two at a time, in all possible combinations showed that from one cellular fraction to another, the radioactivity peaks corresponding to the approximately 70-, 43-45-, and 30-kDa proteins were not superimposed . Their relative positions on the gel differed by one or two slices, which indicated differences in molecular mass of 1.9-3.6 kDa . Moreover, enzymatic digestion with Staphylococcus aureus V8 protease of these three proteins, prepared from each subcellular fraction, yielded dissimilar peptide patterns.(ABSTRACT TRUNCATED AT 250 WORDS) J Biol Chem, 1985 Sep 5, 260(19), 10671 - 9 Amino acid sequence of a proline-rich phosphoglycoprotein from parotid secretion of the subhuman primate Macaca fascicularis; Oppenheim FG et al.; The complete amino acid sequence of the macaque proline-rich phosphoglycoprotein (MPRP) was determined by automated Edman degradation of the protein, fragments F-1 and F-2 derived from the protein by an intrinsic salivary protease, and chymotryptic, tryptic, Staphylococcus aureus V8 protease, and endoproteinase lysine-C peptides . MPRP contains 115 amino acid residues including phosphorylated serine at residues 1, 2, 6, 12, and 15, and 6 O-glycosidic carbohydrate units at residues 69, 75, 87 (threonine) and 96, 103, and 106 (serine) . The Mr of the polypeptide moiety of the protein is 12,656 . The amino-terminal domain contains all 5 phosphoserine residues and most of the other negatively charged and hydrophilic residues, whereas the carboxyl-terminal domain contains 24 of 25 proline residues, and 6 O-glycosidic oligosaccharides . Comparison of MPRP with the four major anionic proline-rich proteins (PRPs) from human glandular secretion shows that 57% of the amino acid residues are identical if gaps are introduced to maximize homology, suggesting that these proteins are phylogenetically related . Significant structural and functional differences occur between the macaque and human proteins . MPRP has 5 phosphoserines, PRPs have 2 . MPRP is a glycoprotein, PRPs are not . MPRP inhibits the spontaneous precipitation (primary precipitation) of calcium phosphate salts from supersaturated solutions in addition to inhibiting seeded crystal growth (secondary precipitation) (Oppenheim, F . G., Offner, G . D., and Troxler, R . F . (1982) J . Biol . Chem . 257, 9271-9282), whereas PRPs inhibit only secondary precipitation . MPRP is the only major anionic proline-rich protein in macaque glandular secretion; in contrast, there are four major anionic PRPs and these display a genetic polymorphism . The significance of these structural differences with respect to biological function and the possible relationship of MPRP to salivary mucins are discussed. J Biol Chem, 1985 Sep 5, 260(19), 10748 - 60 Rat hepatic cytosolic phosphoenolpyruvate carboxykinase (GTP) . Structures of the protein, messenger RNA, and gene; Beale EG et al.; The primary structure of the messenger RNA coding for cytosolic phosphoenolpyruvate carboxykinase was determined by sequencing cDNA and genomic DNA and by primer extension of the mRNA . The molecule is 2624 nucleotides in length; this includes 143 nontranslated nucleotides at the 5' end and 615 nontranslated nucleotides at the 3' end . The 3' nontranslated sequence contains a 102-base pair region of alternating purine-pyrimidine nucleotides (the majority of which are UpG dinucleotides), several direct repeats and palindromic sequences, and 8 CpG dinucleotides . The corresponding segment of the phosphoenolpyruvate carboxykinase gene thus has characteristics which favor the formation of Z-DNA . The amino acid sequence of phosphoenolpyruvate carboxykinase was deduced from the mRNA sequence and confirmed by fast atom bombardment mass spectrometric analysis of peptides generated with trypsin and Staphylococcus aureus V8 protease . The protein consists of 621 amino acids and has a molecular weight of 69,289 . Charon 4A lambda bacteriophage clones containing genomic DNA coding for phosphoenolpyruvate carboxykinase were isolated from a library of partial HaeIII digests of rat liver DNA . Two clones, lambda PC112 and lambda PC103, contained the entire coding region in 15-kilobase inserts and were used to subclone the gene into pBR322 as EcoRI, BamHI, or SstI-KpnI fragments . Using these subclones, the structure of the phosphoenolpyruvate carboxykinase gene was determined by S1 nuclease mapping, R-loop analysis, and DNA sequencing . The gene is composed of 10 exons and 9 introns with a total length of 6.0 kilobases . The transcription initiation site of the gene was determined by a combination of in vitro transcription in a HeLa cell lysate system, primer extension of mRNAPEPCK, and S1 nuclease mapping . In vitro transcription of purified DNA templates revealed three RNA polymerase II-dependent start sites . Two sites were separated by 600 base pairs on the coding strand and the third site was on the noncoding strand . The products of S1 nuclease mapping and primer extension from a BglII site were compared in order to determine which of the coding strand initiation sites was expressed in vivo . In both cases a 69-base pair fragment was generated and the 5' end of this corresponded to a thymidine residue identified in a sequence ladder of the genomic DNA coding strand . We conclude that mRNAPEPCK synthesis initiates with an adenine residue 69 base pairs 5' of the BglII site; this corresponds to the 3' most transcription initiation site determined in vitro. J Immunol Methods, 1985 Sep 3, 82(1), 131 - 40 A one-plate assay for macrophage bactericidal activity; Peck R; Human peripheral blood monocytes purified by counterflow centrifugal elutriation (CCE) showed an enhanced capacity to kill Listeria Monocytogenes and Staphylococcus aureus after activation with interferon-gamma (IFN gamma) . Bactericidal activity was evaluated by a rapid colorimetric microassay based on the reduction of a tetrazolium dye, MTT . The amount of dye reduced was directly proportional to the number of viable bacteria present in each microwell . Comparison with a bacterial titration plate containing serial dilutions from stock of a known titer allowed calculation of bacterial numbers before and after exposure to macrophages . Results were read in a multiscan plate reader and compared closely with those obtained by serial dilution on agar plates (i.e., colony counts) . The entire assay, i.e., culturing the monocytes, treatment with IFN gamma, addition of the bacteria, and quantitation of surviving viable bacteria, was performed in a single microtiter plate. J Biochem (Tokyo), 1985 Sep, 98(3), 687 - 94 The multiplicity of human pancreatic secretory trypsin inhibitor; Kikuchi N et al.; Four forms of pancreatic secretory trypsin inhibitor (PSTI; A1, A2, B, and C) were purified from human pancreatic juice . According to sequence results, the primary structure of B was different from that reported earlier (Greene, L.J., et al . (1976) Method Enzymol . 45, 813-825) at two positions, i.e . Asn21----Asp21, Asp29----Asn29 . A1 and A2 were deamidated forms of B judging from peptide mappings with Staphylococcus aureus V8 protease . Gln45 in B was replaced by Glu in A1 and Gln51 in B was replaced by Glu in A2 . C was an inhibitor lacking five amino acid residues from the amino terminal of B . B and C inhibited human cationic trypsin activity stoichiometrically with similar dissociation constants, but A1 and A2 showed poorer trypsin inhibitory activity than B and C. Appl Environ Microbiol, 1985 Sep, 50(3), 696 - 7 Transport and processing of staphylococcal enterotoxin A; Christianson KK et al.; A larger-molecular-weight precursor of enterotoxin A was found in membranes of Staphylococcus aureus and was shown to be the kinetic precursor to the extracellular form of the toxin . Subcellular fractionation revealed that mature enterotoxin A was transiently associated with the cell wall before being released to the extracellular environment. Antimicrob Agents Chemother, 1985 Sep, 28(3), 456 - 7 Oscillating tolerance in synchronized cultures of Staphylococcus aureus; Holzhoffer S et al.; Cells of synchronized cultures of Staphylococcus aureus showed an oscillating MBC/MIC ratio when tested with penicillin G . Although the MICs did not differ significantly throughout the cell cycle, the MBC was at its maximum when actively dividing cells were inoculated. J Antimicrob Chemother, 1985 Sep, 16(3), 335 - 9 Effects of clindamycin in combination with rifampicin on clindamycin-susceptible and clindamycin-resistant Staphylococcus aureus; Watanakunakorn C; The effects of the combination of clindamycin and rifampicin against 21 strains of clindamycin-susceptible and 19 strains of clindamycin-resistant Staphylococcus aureus were studied by the time-kill method . For the clindamycin-susceptible strains, clindamycin prevented the re-growth of Staph . aureus in the presence of rifampicin . For the clindamycin-resistant Staph . aureus strains, indifference was demonstrated against the majority of strains, with only a few strains showing synergism or antagonism. J Clin Microbiol, 1985 Sep, 22(3), 452 - 4 Nasal carriage of Staphylococcus aureus and antistaphylococcal immunoglobulin E antibodies in atopic dermatitis; Falanga V et al.; Twelve atopic dermatitis patients were studied to investigate the relationship between levels of antistaphylococcal immunoglobulin E antibodies in serum and quantitative cultures of Staphylococcus aureus strains from the anterior nares and chronic lesions . A positive correlation was found between logarithmic counts of S . aureus strains from the anterior nares and levels of antistaphylococcal immunoglobulin E in serum . The observation is important for understanding the pathophysiology of atopic dermatitis. Infect Immun, 1985 Sep, 49(3), 765 - 9 Virulence of Staphylococcus aureus in a mouse mastitis model: studies of alpha hemolysin, coagulase, and protein A as possible virulence determinants with protoplast fusion and gene cloning; Jonsson P et al.; Mutants of a genetically well-characterized strain of Staphylococcus aureus {SA113(83A)} were isolated after mutagenization . Alpha-hemolysin- (hla), coagulase- (coa), and protein A- (spa) negative mutants were characterized by more than 90 biochemical tests for production of extracellular proteins and biochemical profile to exclude pleiotropy . Protoplast fusion was then used to isolate double-defective (hla and coa) recombinants and recombinants with regained properties, i.e., production of alpha-hemolysin and coagulase . Studies of such mutants and recombinants in the mouse mastitis model showed that one alpha-hemolysin {SA113(83A) hla-5} and one coagulase-negative {SA113(83A) coa-147} mutant were lower in virulence compared with the wild-type strain SA113(83A) . The double-negative mutant SA113(83A) hla-5 coa-147 showed a drastic decline in virulence and only induced very mild changes, as determined by microscopic examinations of infected mammary gland tissue . The recombinant with regained properties, however, was as virulent as the wild-type strain . This suggests that alpha-hemolysin and coagulase are virulence determinants of S . aureus . A high-level protein A-producing mutant (U300) showed the same virulence as the parent strain SA113(83A) in this model . One low virulence protein A-negative mutant (U320) did not markedly increase in virulence when a plasmid containing the cloned gene for protein A (pSPA15) was introduced into this mutant . By these and earlier observations, it seems likely that protein A is not an important virulence determinant in mastitis of mice . The reduced virulence of the protein A-negative mutant U320 compared with the wild-type SA113(83A) may be due to pleiotropic loss of some other unknown virulence determinant(s) . Our data confirm earlier findings that pleiotropic changes are common in protein A-negative mutants. South Med J, 1985 Sep, 78(9), 1140 - 1 Pacemaker-associated sepsis: successful medical management; Tylman TA et al.; We have presented a case of Staphylococcus aureus septicemia in a patient with a transvenous pacemaker who responded to conservative medical therapy. J Infect Dis, 1985 Sep, 152(3), 521 - 8 Interaction between human monocytes and penicillin G in relation to the antibacterial effect on Staphylococcus aureus; van den Broek PJ et al.; The influence of the presence of monocytes on the effect of penicillin G on Staphylococcus aureus was studied . Conditions were varied in such a way that phagocytosis and intracellular killing of S . aureus by monocytes did not occur, was short lasting and limited, or long lasting and extensive . Synergism between penicillin G and monocytes was observed in the absence of and during limited phagocytosis and intracellular killing by the monocytes . When phagocytosis and intracellular killing were optimal, addition or antagonism between penicillin G and monocytes was observed . Enhancement of the antibacterial effect of penicillin G was also obtained in the presence of monocyte supernatant (the cell-free medium in which monocytes had been incubated for 3 hr) . This indicates that monocytes secrete a factor that enhances the antibacterial activity of penicillin G . Serum inhibits the activity of this factor. Clin Orthop, 1985 Sep, (198), 231 - 9 Methicillin-resistant Staphylococcus aureus osteomyelitis; Sheftel TG et al.; In five patients, the diagnosis of methicillin-resistant Staphylococcus aureus (MRSA) osteomyelitis was made by clinical and roentgenographic methods and confirmed by bone biopsy cultures . The treatment was staged according to the anatomic setting of the infection and the systemic and local competence of the host . Seven episodes of osteomyelitis were encountered in the five patients . Two patients had persistence of their infection and were successfully treated by additional surgical debridement, antibiotics, and adjunctive hyperbaric oxygen . Vancomycin was administered to all patients . The daily dosage of vancomycin ranged between 100 mg and 2.0 gm . The length of vancomycin therapy ranged from 19 to 56 days . Five of seven biopsy specimens grew bacterial organisms in addition to MRSA . The MIC of vancomycin for MRSA ranged between 0.39 and 1.56 micrograms/ml . Osteomyelitis was arrested in five of seven episodes, and follow-up evaluations ranged from two to 35 months . Two of five (40%) patients receiving the combination of vancomycin and tobramycin developed signs of renal toxicity . Renal function returned to normal after discontinuation of the antibiotics . MRSA osteomyelitis is usually acquired by spread from a contiguous focus of infection and is often polymicrobic in nature . Treatment with vancomycin or vancomycin plus tobramycin when the infection was polymicrobic was effective . The combination of vancomycin plus tobramycin is potentially nephrotoxic. Appl Environ Microbiol, 1985 Sep, 50(3), 634 - 7 Application of cross-linked carboxymethyl cellulose degradation by beta-glucosidase and vaginal microbes to toxic shock syndrome; Sierks MR et al.; Eleven bacterial and two yeast strains, four of which were previously identified as having activity on a lightly cross-linked carboxymethyl cellulose (CLD-2) found in one type of superabsorbent tampon, were grown on a variety of substrates, most containing cellulosics . None produced detectable amounts of cellulases, but all elaborated beta-glucosidase . None of these 13 strains nor 3 commercially obtained beta-glucosidase preparations could hydrolyze CLD-2, although a commercial cellulase and two other bacterial preparations known to produce cellulases could . Based on these results, it appears that previous work suggesting that the degradation of CLD-2 by vaginal microbes and beta-glucosidase is implicated in the production by Staphylococcus aureus of toxin causing toxic shock syndrome must be reevaluated. J Clin Microbiol, 1985 Sep, 22(3), 459 - 61 Misidentification of mucoid variants of Staphylococcus aureus by standard laboratory techniques; Notarnicola SM et al.; Ten atypical strains of Staphylococcus aureus were examined and compared with previously characterized S . aureus strains . Five of the strains were sent to two hospital microbiology laboratories for identification . All of the mucoid strains were misidentified when tested by standardized coagulase, DNase, and mannitol fermentation criteria, as well as by an automated identification method, but were accurately identified by the API system (Analytab Products, Plainview, N.Y.) . These results illustrate the possibility of misidentification when reliance is placed on a few key characteristics rather than a thorough characterization of unusual Staphylococcus isolates. J Clin Microbiol, 1985 Sep, 22(3), 445 - 7 Method for the serological typing of the capsular polysaccharides of Staphylococcus aureus; Karakawa WW et al.; A method is described for typing Staphylococcus aureus capsular polysaccharides that is based on direct bacterial cell agglutination and immunoprecipitation of cell extracts with monospecific antisera . Encapsulated strains were identified by their inagglutinability with teichoic acid antisera . The typing sera reacted specifically with extracts of eight prototype strains. Arch Biochem Biophys, 1985 Sep, 241(2), 577 - 89 Complete amino acid sequence of copper-zinc superoxide dismutase from Drosophila melanogaster; Lee YM et al.; The complete amino acid sequence of the Drosophila melanogaster Cu,Zn superoxide dismutase subunit has been determined by automated Edman degradation . Sequence analyses were performed on the intact S-carboxymethylated protein, two fragments derived from CNBr cleavage, and three peptides recovered from mouse submaxillary protease digestion of the reduced and S-carboxymethylated enzyme . The peptides were aligned by characterizing peptides yielded by trypsin and Staphylococcus aureus V8 protease . All the peptides studied were purified exclusively by reverse-phase columns of HPLC and were analyzed with an improved liquid-phase sequencer . A molecular weight of 15,750 (subunit) was calculated from the 151 residues sequenced . The amino acid sequence of the Drosophila superoxide dismutase subunit is compared with that of four other eucaryotes: man, horse, cow, and yeast . Comparison of the five primary structures reveals very different rates of evolution at different times . Copper-zinc superoxide dismutase appears to be a very erratic evolutionary clock . Val-Val-Lys-Ala- Val-Cys-Val-Ile-Asn-Gly-Asp-Ala-Lys-Gly-Thr-Val-Phe-Phe-Glu-Gln- Glu-Ser-Ser-Gly-Thr-Pro-Val-Lys-Val-Ser-Gly-Glu-Val-Cys-Gly-Leu- Ala-Lys-Gly-Leu-His-Gly-Phe-His-Val-His-Glu-Phe-Gly-Asp-Asn-Thr- Asn-Gly-Cys-Met-Ser-Ser-Gly-Pro-His-Phe-Asn-Pro-Tyr-Gly-Lys-Glu- His-Gly-Ala-Pro-Val-Asp-Glu-Asn-Arg-His-Leu-Gly-Asp-Leu-Gly-Asn- Ile-Glu-Ala-Thr-Gly-Asp-Cys-Pro-Thr-Lys-Val-Asn-Ile-Thr-Asp-Ser- Lys-Ile-Thr-Leu-Phe-Gly-Ala-Asp-Ser-Ile-Ile-Gly-Arg-Thr-Val-Val-Val- His-Ala-Asp-Ala-Asp-Asp-Leu-Gly-Gln-Gly-Gly-His-Glu-Leu-Ser-Lys- Ser-Thr-Gly-Asn-Ala-Gly-Ala-Arg-Ile-Gly-Cys-Gly-Val-Ile-Gly-Ile- Ala-Lys. J Cell Biol, 1985 Sep, 101(3), 824 - 9 Cathepsin D precursors in clathrin-coated organelles from human fibroblasts; Schulze-Lohoff E et al.; Coated vesicles were isolated from metabolically labeled human fibroblasts with the aid of affinity-purified antibodies against human brain clathrin and Staphylococcus aureus cells . The material adsorbed to the S . aureus cells was enriched in clathrin . When the S . aureus cells bearing the immunoadsorbed material were treated with 0.5% saponin, extracts containing the precursor form of cathepsin D were obtained . The extraction of the precursor was promoted in the presence of mannose 6-phosphate . Material adsorbed to S . aureus cells coated with control immunoglobulins was nearly free of clathrin and contained a small amount of the cathepsin D precursor (less than 20% of that adsorbed with anti-clathrin antibodies) . The extraction of this cathepsin D precursor was independent of mannose 6-phosphate and was complete after a brief exposure to saponin . The amount of cathepsin D precursor in coated membranes varied between 0.4 and 2.5% of total precursor . Analysis of pulse chase-labeled fibroblasts revealed that cathepsin D was only transiently associated with coated membranes . The mean residence time of cathepsin D precursor in coated membranes was estimated to be 2 min . These observations support the view that coated membranes participate in the transfer of precursor forms of endogenous lysosomal enzymes to lysosomes. Anal Biochem, 1985 Sep, 149(2), 322 - 30 Peptide mapping of bovine pancreatic ribonuclease A by reverse-phase high-performance liquid chromatography . II . A two-dimensional technique for determination of disulfide pairings using a continuous-flow disulfide-detection system; Thannhauser TW et al.; A procedure, developed for the cleavage and reversible blocking of disulfide bonds of proteins by S-sulfonation in preparation for peptide mapping, was applied to ribonuclease A . The complete peptide maps of sulforibonuclease A using limited Staphylococcus aureus protease digestion, tryptic digestion, and tryptic followed by chymotryptic digestion are presented . A description is given of an adaptation of the sulfonation procedure which forms the basis of a sensitive (5-pmol detection limit) and quantitative (+/- 5%) disulfide-detection system for the continuous monitoring of HPLC column effluents for disulfide-containing compounds . The sulfonation procedure, peptide maps, and disulfide-detection system are the key ingredients in a two-dimensional reverse-phase HPLC technique for the determination of disulfide pairings . The applicability of this technique is demonstrated by determining the known disulfide pairings of ribonuclease A . It is also shown that there is no disulfide interchange under the digestion conditions used . This technique is suitable for determining the distributions of disulfide pairings in the intermediates present in the oxidative folding of disulfide-containing proteins. Plasmid, 1985 Sep, 14(2), 152 - 61 Ribosomal RNA methylation in Staphylococcus aureus and Escherichia coli: effect of the "MLS" (erythromycin resistance) methylase; Thakker-Varia S et al.; Classical acquired resistance to erythromycin in Staphylococcus aureus ("MLS," or macrolide-lincosamide-streptogramin, resistance) was shown by Weisblum and colleagues to be a direct consequence of the conversion of one or more adenosine residues of 23S rRNA, within the subsequence(s) GA3G, to N6-dimethyladenosine (m62A) . The methylation reaction is effected by a class of methylase, whose genes are typically plasmid- or transposon-associated, and whose synthesis is inducible by erythromycin . Using a recently obtained clinical MLS isolate of S . aureus, we have further defined the methylation locus as YGG X m62A X AAGAC; and have shown that this subsequence occurs once in the 23S RNA and that it is essentially completely methylated in all copies of 23S RNA that accumulate in induced cultures . Similar findings were obtained with laboratory S . aureus strains containing two well-characterized evolutionary variants (ermB, ermC) of MLS methylase genes . Analyses of a strain of E . coli containing the ermC gene indicated that the specificity of the methylase gene was unchanged, but that its expression was muted . Even after prolonged periods of induction, the strain manifested only partial resistance to erythromycin, and only about one-third of the copies of the MLS subsequence were methylated in such "induced" cultures . Since the E . coli 23S RNA sequence is known in its entirety, localization of the MLS subsequence is in this case unambiguous; as inferred by homology arguments applied earlier to the S . aureus data, the subsequence is in a highly conserved region of 23S RNA considered to contribute to the peptidyl transferase center of the ribosome. Antibiot Med Biotekhnol, 1985 Sep, 30(9), 684 - 7 {Potentiation of the action of antibiotics by ultrasound}; Ukhov AIa et al.; The aim of the study was to investigate the action of ultrasound on antimicrobial drugs and to estimate the effect of antibacterial drugs and ultrasound used in combination and alone . In the first series of the experiments it was shown that under the action of ultrasound such antibiotics as benzylpenicillin, streptomycin, ampicillin, lincomycin, monomycin, rifampicin and gentamicin, and antiseptic drugs such as furacin, rivanol and iodinol did not change their antibacterial (bacteriostatic and bactericidal) properties with regard to Staphylococcus aureus and Escherichia coli . In the second series of the experiments it was shown that ultrasound potentiated by 1.7-10.8 times the antibacterial effect of the drugs . Probably, ultrasound promoted closer contacts of the drugs with the microbial cell . In the third series of the experiments it was shown that ultrasound lowered to 24.1 per cent the count of the bacterial cells capable of multiplication, while ampicillin lowered their count to 52.6 per cent . When the antibiotic and ultrasound were used in combination the count of the bacterial cells capable of multiplication was lowered to 5.2 per cent . Therefore, the combined use of the antibacterial drug and ultrasound resulted in a 4.48-fold increase of the total effect. Ann Ophthalmol, 1985 Sep, 17(9), 560 - 4 Influence of drug vehicle on ocular contact time of sulfacetamide sodium; Hanna C et al.; Sulfacetamide sodium drops and ointments were applied to the conjunctiva of patients and volunteers . Tear samples were taken and analyzed for sulfacetamide sodium content . The minimal inhibitory concentrations of the drug in para-aminobenzoic acid-free media were determined to be between 20 and 50 micrograms/mL against Escherichia coli and Staphylococcus aureus, and the effective concentrations of the sulfa were taken as 50 micrograms/mL . The concentration of sulfacetamide sodium in tears fell to the 50 micrograms/mL level in 30 minutes, two hours, and five and a half hours after the single application of 10 microL of 15% and 30% drops and 25 microL of all three ointments, respectively . The ocular contact time of the sulfa was increased by increasing the concentration of drug . The makeup of the three ointments differed greatly, yet the ocular contact time was similar. Virus Res, 1985 Sep, 3(2), 153 - 63 In vitro translation of the major capsid polypeptide from Ustilago maydis virus stain P1; Dalton RE et al.; Double-stranded RNA (dsRNA) from Ustilago maydis virus strain P1 was translated in vitro using a nuclease-treated rabbit reticulocyte lysate system . Following heat denaturation of the H2 double-stranded RNA segment in 90% dimethyl sulfoxide and incubation in the cell free extract, a primary translation product was observed which showed the same molecular weight and co-migrated with viral coat protein on 10% SDS-polyacrylamide gels . The in vitro product of the H2 dsRNA segment could also be immunoprecipitated with antibodies prepared against viral coat protein . Limited proteolysis of the in vitro product and authentic viral coat protein using Staphylococcus aureus V8 protease produced similar peptide patterns on SDS gels . In vitro translation products from other dsRNA segments that make up the P1 viral genome could not be precipitated by antibody to viral coat protein . These results complement the genetic data that indicated that information for coat formation and maintenance was contained within the H segments of dsRNA. J Antimicrob Chemother, 1985 Sep, 16(3), 349 - 58 Penetration of beta-lactam antibiotics into cardiac vegetations, aorta and heart muscle in experimental Staphylococcus aureus endocarditis: comparison of ceftazidime, cefuroxime and methicillin; McColm AA et al.; The penetration of ceftazidime, cefuroxime and methicillin into cardiac vegetations, normal aorta wall tissue, heart muscle and heart muscle tissue fluid was measured in rabbits with Staphylococcus aureus (853E) endocarditis . After a 100 mg/kg intramuscular injection, ceftazidime attained significantly higher peak concentrations than cefuroxime and methicillin in all compartments with the exception of aorta where no difference was observed between ceftazidime and methicillin . The half life of ceftazidime in each compartment (50-65 min) was approximately twice that of cefuroxime and methicillin . The area under the concentration/time curve for ceftazidime in vegetations over a post dosing 6 h period was approximately five times greater than that for cefuroxime and three times that for methicillin . Ceftazidime remained for a longer time period in all compartments, however cefuroxime persisted the longest at supra-MIC concentrations in all compartments except serum . Compared to cefuroxime and methicillin where vegetation and aorta tissue levels were similar, ceftazidime attained peak levels in vegetations which were more than double those found in aorta tissue . Analysis of the percentage penetration from serum into vegetation, suggested that the higher concentrations of ceftazidime in vegetation tissue were probably not a function of increased fluid content relative to undamaged aorta but were more likely caused by an intrinsically better penetrating capacity for vegetation tissue. Proc Natl Acad Sci U S A, 1985 Sep, 82(17), 5850 - 4 Molecular cloning of staphylococcal enterotoxin B gene in Escherichia coli and Staphylococcus aureus; Ranelli DM et al.; We have cloned the Staphylococcus aureus entB gene in Escherichia coli, using pBR322 as the vector plasmid; however, no detectable staphylococcal enterotoxin B (SEB) was produced by the E . coli clones . When the entB gene was placed downstream from the strong lambda phage promoter, PR, SEB was synthesized at readily detectable levels in E . coli . Interestingly, mature SEB was almost exclusively present in the cytoplasmic fraction . The SEB precursor was found associated with the cell membrane . The entB gene was introduced back into S . aureus, and the clones were shown to produce SEB . The entB gene has been located to a 2.1-kilobase-pair region . Maxam-Gilbert sequencing of a part of the entB gene yielded a DNA sequence that corresponds to the known amino acid sequence of SEB . Southern hybridization experiments showed that the entB gene was present on identical restriction fragments in the chromosomes of SEB-producer strains . The entB gene is absent from SEB-nonproducer strains. Proc Natl Acad Sci U S A, 1985 Sep, 82(17), 5732 - 6 Amino acid sequence of the heat-stable inhibitor of the cAMP-dependent protein kinase from rabbit skeletal muscle; Scott JD et al.; The amino acid sequence of rabbit skeletal muscle heat-stable inhibitor of the cAMP-dependent protein kinase has been determined by microsequencing techniques . Proof of the structure involved a series of nonoverlapping tryptic fragments for primary identification of 86% of the amino acids . Complementary fragments generated by cleavage with chymotrypsin, Staphylococcus aureus V8 proteinase, and mast cell proteinase II contributed to proof of the structure . The inhibitor is a single polypeptide chain of 75 residues and has a molecular weight of 7829 . It lacks tryptophan, proline, and sulfur-containing amino acids . The amino terminus of the inhibitor is blocked by an unidentified group . The amino-terminal region of the molecule contains the kinase inhibitory domain, and synthetic peptides based on the sequence of residues 11-30 are potent competitive inhibitors of the cAMP-dependent protein kinase {Scott, J . D., Fischer, E . H., Demaille, J . G . & Krebs, E . G . (1985) Proc . Natl . Acad . Sci . USA 82, 4379-4383} . Residues 14-22 show considerable homology to the "hinge-regions" of the regulatory subunits of the cAMP-dependent protein kinase . The remainder of the molecule shows no similarity to the known amino acid sequence of any protein. Helv Paediatr Acta, 1985 Sep, 40(4), 277 - 84 Surgery and granulocyte transfusions for life-threatening infections in chronic granulomatous disease; Fanconi S et al.; We report two patients with chronic granulomatous disease (CGD) and life-threatening infections: a 10 10/12-year-old boy had Aspergillus fumigatus spondylitis with destruction of the 11th vertebral body and paravertebral abscess formation, and an 8 5/12-year-old boy had multiple Staphylococcus aureus hepatic abscesses with subphrenic abscess formation . Both patients failed to respond to intense antimicrobial therapy but showed a remarkable recovery following surgical drainage combined with granulocyte transfusions . These results suggest that antimicrobial therapy and surgical drainage followed by granulocyte transfusions may be the ideal mode of treatment for severe infections in patients with CGD. Antimicrob Agents Chemother, 1985 Sep, 28(3), 397 - 403 Role of an altered penicillin-binding protein in methicillin- and cephem-resistant Staphylococcus aureus; Utsui Y et al.; About 80% of methicillin- and cefazolin-resistant strains of Staphylococcus aureus isolated clinically in Japan in 1982 retained their resistance even after elimination of penicillinase-encoding plasmids . The penicillin-binding proteins (PBPs) of the penicillinase-free, methicillin- and cephem-resistant subclones of Staphylococcus aureus (MRSA) were compared with those of spontaneous susceptible revertants which had been obtained by the replica method after 10 subcultures in drug-free media . A new PBP fraction (PBP2') having a molecular weight of 78,000 and low binding affinities for various beta-lactam antibiotics was found in MRSA exclusively . The levels of resistance of MRSA strains were reduced markedly by culturing them at 43 degrees C or at pH 5.2 or both . We found that the binding capacity of PBP2' for 14C-labeled penicillin G was decreased by preincubation of the membrane fractions of MRSA strains at 43 degrees C for 60 min and that the amount of PBP2' in MRSA strains grown at pH 5.2 was less than that the amount of PBP2' in MRSA strains grown at pH 7.0 . Temperature- and pH-dependent expression of resistance in MRSA is likely to reflect the temperature sensitivity and neutral pH-dependent production of the specific PBP fraction (PBP2') . We suggest that MRSA strains can grow in the presence of beta-lactam antibiotics because of the low affinities of the specific PBP2' fraction for various beta-lactam antibiotics. Zh Mikrobiol Epidemiol Immunobiol, 1985 Sep, (9), 77 - 80 {Dynamics of staphylococcal infection in guinea pigs with delayed hypersensitivity to Staphylococcus aureus surface antigens}; Pashutin SB et al.; The relationship between delayed hypersensitivity (DH) to S . aureus surface antigens and the intensity of the infectious process induced by the sublethal infection of guinea pigs with S . aureus was studied . The protective effect, manifested by a decrease in the staphylococcal contamination of the spleen tissue and by an increase in the level of the activation of lymphocytes, was shown to correlate with DH induced by inactivated staphylococcal cells . In infected guinea pigs having DH to different staphylococcal antigens the disease either took a more severe course (in cases of DH to cell wall or peptidoglycan) than in the animals subjected only to infection, or no aggravation of the disease was observed (in cases of DH to protein A). Pharmazie, 1985 Sep, 40(9), 650 - 1 Combined action of amoxycillin and dicloxacillin against Staphylococcus aureus in vitro; Yousef RT et al.; Combination of amoxycillin (1) and dicloxacillin (2) showed synergistic bacteriostatic and bactericidal activities against 15 clinical isolates of Staphylococcus aureus, including 3 beta-lactamase-producing strains. Biochem J, 1985 Sep 1, 230(2), 353 - 61 Amino acid sequence of the trypsin-generated C3d fragment from human complement factor C3; Hellman U et al.; Human C3d (try-C3d), prepared from trypsin-digested C3, was fragmented by cleavage with CNBr . Eight peptides were defined and separated by h.p.l.c . on reversed-phase columns . By automatic Edman degradation the complete sequences of five peptides and partial sequences of three peptides were determined . To obtain overlapping peptides the latter three fragments were digested with trypsin, chymotrypsin or Staphylococcus aureus V8 proteinase, after which the fragments were separated on reversed-phase columns . Two of the CNBr-cleavage peptides were completely sequenced, and 70% of the sequence of the remaining CNBr-cleavage peptide was determined . The non-sequenced part represents a very hydrophobic segment of try-C3d . The sequence data obtained represent 90% of the primary structure of try-C3d . Alignment of the CNBr-cleavage fragments was made easier by comparison with the cDNA sequence of mouse pro-C3 {Wetsel, Lundwall, Davidson, Gibson, Tack & Fey (1984) J . Biol . Chem . 259, 13857-13862} . Comparison of try-C3d with the equivalent part of human C4B revealed an extensive sequence homology in the N-terminal half of the molecules. Arthritis Rheum, 1985 Sep, 28(9), 1008 - 15 Impaired B cell proliferation by Staphylococcus aureus Cowan 1 in patients with systemic lupus erythematosus; Sawada S et al.; We examined the proliferative response to Staphylococcus aureus Cowan 1 (SAC) by enriched peripheral blood B cells from patients with systemic lupus erythematosus (SLE) . Responses of B cells from patients with active and inactive SLE were significantly lower than those of B cells from normal individuals . Hyporesponsiveness to SAC was not observed in healthy family members of SLE patients . This hyporesponsiveness did not correlate with prednisolone therapy and could not be attributed to serum factors; it did correlate with the presence of suppressor monocytes . However, we could not exclude the possibility of enhanced sensitivity of SLE B lymphocytes to suppressive signals delivered by the monocytes. Infect Immun, 1985 Sep, 49(3), 664 - 9 C1q, a subunit of the first component of complement, enhances binding of plasma fibronectin to bacteria; Sorvillo JM et al.; The interaction of plasma fibronectin with C1q of the complement system has been demonstrated in the past several years . In addition, the antibody-independent binding of C1q to bacteria, as well as the binding of plasma fibronectin to bacteria, is well documented . This study examines whether the binding of C1q to bacteria enhances the interaction of C1q and bacteria with plasma fibronectin . Highly purified 125I-C1q bound to several species of bacteria in the absence of antibody . The binding of 125I-C1q to bacteria was saturable and specific since the addition of unlabeled C1q inhibited binding while the presence of bovine serum albumin did not . Bacteria which had been pretreated with either buffer or unlabeled C1q were tested for their ability to bind 125I-fibronectin . When bacteria were preincubated with buffer, Staphylococcus aureus bound fivefold more 125I-fibronectin than did Escherichia coli . However, preincubation of E . coli with C1q increased the binding of 125I-fibronectin by up to 20-fold, whereas pretreatment of S . aureus with C1q increased fibronectin binding by only twofold . These results were confirmed by immunoblotting studies which demonstrated the presence of C1q, as well as an increase in fibronectin antigens on the C1q-treated bacteria as compared with the level of fibronectin on buffer-treated bacteria . In addition, preincubation of 3H-labeled bacteria with C1q enhanced their attachment to fibronectin-coated surfaces but not to albumin-coated surfaces . The biological consequences of these observations are discussed. EMBO J, 1985 Sep, 4(9), 2295 - 300 Regulation of the inducible chloramphenicol acetyltransferase gene of the Staphylococcus aureus plasmid pUB112; Bruckner R et al.; Analyses of deletion mutants of the gene for chloramphenicol (Cm) acetyltransferase (CAT) carried by the staphylococcal plasmid pUB112 revealed a regulatory region, which is indispensable for Cm-inducible cat gene expression, located 70 bp in front of the CAT-coding sequence . This region consists of a possible ribosome binding site followed by an open reading frame coding for a peptide of nine amino acids and overlaps partially with an inverted repeat capable of forming a stem-loop structure . Deletion of the ribosome binding site and of parts of the open reading frame abolishes inducibility and results in a low-level cat gene expression, if the inverted repeat remains intact . Deletion of the 5' part of the possible stem leads to high-level constitutive CAT synthesis . The inverted repeat, therefore, exhibits negative control on cat gene expression whereas the preceding ribosome binding site is needed to enhance CAT synthesis in the presence of an inducer . These results suggest that translation of a leader peptide is a prerequisite for Cm-induced cat gene expression and that ribosome stalling on cat leader mRNA caused by Cm opens the stem-loop structure thereby releasing its negative effect on CAT synthesis. Antimicrob Agents Chemother, 1985 Sep, 28(3), 381 - 8 Evaluation of oxacillin tolerance in Staphylococcus aureus by a novel method; Woolfrey BF et al.; A novel agar dilution plate-count procedure for the quantitative measurement of bacterial inhibition and killing is described . For Staphylococcus aureus versus oxacillin, by the agar dilution plate-count procedure it was found that only 1 of 20 clinical isolates and 1 of 7 allegedly tolerant reference isolates met the conventional definition of tolerance . By using inocula of 10(5) CFU per plate, most isolates were demonstrated to have subpopulations of cells which, although inhibited, persisted for 24 h in concentrations significantly above their MICs . The persister percentages at 24 h appeared to be strain dependent, and all persisters exhibited the paradoxical effect . For each isolate, the persister percentage markedly decreased after action by oxacillin for 48 h, and the paradoxical effect was greatly diminished . Our findings suggest that tolerance is an artificial and arbitrary concept that does not adequately characterize the inhibition and killing dynamics associated with the persister phenomenon. J Clin Microbiol, 1985 Sep, 22(3), 467 - 9 Use of the Autobac system for detection of methicillin-resistant Staphylococcus aureus; de Rautlin de la Roy Y et al.; Using 490 strains of Staphylococcus aureus divided into methicillin-susceptible, -resistant, and -heteroresistant varieties, we compared the results obtained by the agar disk method with those obtained with the automated Autobac system . Susceptible strains exhibited a perfect correlation, whereas there were numerous discrepancies with resistant and still more with heteroresistant varieties . When incubation was increased to 18 h at 37 degrees C (Autobac incubation temperature), 35 degrees C, or 30 degrees C, these differences disappeared, but other problems may arise when incubation is prolonged, especially with erythromycin . We thus recommend carrying out two readings, a normal one after 3 h of incubation and a special reading after 18 h, solely for the detection of heteroresistance to methicillin. J Diarrhoeal Dis Res, 1985 Sep, 3(3), 145 - 8 Chronic diarrhoea in Nigerian children; Yakubu AM et al.; PIP: To study the etiology of chronic childhood diarrhea among Nigerian children, 142 patients, aged 6 months to 5 years, with diarrhea for at least 1 month, were evaluated; the study took place during January-December 1983 at the Ahmadu Bello University Teaching Hospital, Zaria, Northern Nigeria . Enteropathogenic agents were identified in stools of 90 (63%) patients . Giardia lamblia and Entamoeba histolytica were most commonly detected, representing 41% and 23%, respectively, of all parasitic pathogens . In children with negative stool microscopy, chronic diarrhea was associated with primary lactose intolerance (2 cases), abdominal tuberculosis (2 cases), hyponatremia, low serum albumin, anemia due to sickle cell disease, or Staphylococcus aureus infection . In contrast with chronic diarrhea etiologies reported among children in Europe and North America, infections were the major cause of chronic childhood diarrhea among these children . In general, it is accepted that intestinal infection usually produces acute diarrhea--and that, if the host fails to mount a competent immune response, if there is repeated exposure to infectious agents, or if severe infection damages a substantial proportion of absorptive cells, then severe, protracted diarrhea may result . The high case fatality rate of 9% in this series was associated with specific infectious complications of septicemia, bronchopneumonia, lobar pneumonia and measles . Severe malnutrition also worsened the prognosis in chronic diarrhea . The results indicate that early detection and treatment of amebiasis and giardiasis is a useful approach in the treatment of chronic diarrhea cases among children . J Gen Virol, 1985 Sep, 66 ( Pt 9), 2017 - 27 The virion proteins and ultrastructure of Staphylococcus aureus bacteriophages; Lee JS et al.; The number and size of the major virion polypeptides have been determined by SDS-PAGE for the 22 Staphylococcus aureus phages of the International Typing Set, plus phages 11 and 80 alpha . Virion ultrastructure was examined by electron microscopy after negative staining with ammonium molybdate . In addition, serogroup B phages were disrupted and fractionated into head, tail-tube and baseplate components and major polypeptides assigned to these substructures . The number and size of the polypeptides correlated closely with the division of aureophages into four serogroups (A, B, F, L), although serogroup L was represented in the set by only a single phage (187) . Apart from serogroup B, however, the polypeptide patterns did not reflect differences between lytic groups . Within serogroup B, polypeptide analysis yielded characteristic patterns for lysogroups I, II and III . Ultrastructural analyses confirm the data provided by polypeptide analysis . Thus, phages from the four serogroups can be identified on the basis of tail-tube length alone, although the differences between phage 187 and members of lysogroups I and III in serogroup B were less than 20 nm, or approximately 12% of the total length . Serogroup A virions differed from those of the other serogroups in that all members of the typing set in this group had elongate, rather than isometric, heads. J Antibiot (Tokyo), 1985 Sep, 38(9), 1246 - 50 Cloning and expression of an APH(3')-III phosphotransferase from Staphylococcus aureus in Streptomyces lividans; Crameri R et al.; An aminoglycoside 3' type III phosphotransferase derived from Staphylococcus aureus plasmid pRN1956 was cloned on the high copy number Streptomycetes vectors pIJ702 and pIJ704 . Streptomyces lividans transformants carrying the hybrid plasmids show a resistance pattern towards aminoglycoside antibiotics comparable to the resistance pattern of S . aureus . The APH(3')-III with expanded spectrum of resistance, is a useful additional marker for gene cloning in Streptomycetes. Eur J Cell Biol, 1985 Sep, 38(2), 312 - 21 Immuno-isolation of vesicles using antigenic sites either located on the cytoplasmic or the exoplasmic domain of an implanted viral protein . A quantitative analysis; Gruenberg J et al.; In this study, we present a new general approach for immuno-isolation: a foreign integral membrane protein, the G-protein of vesicular stomatitis virus (VSV), is implanted into the plasma membrane for subsequent immuno-isolation . A quantitative analysis was accomplished using the erythrocyte plasma membrane as a model system . The virus was artificially bound to the membrane via a lectin and subsequently fused at low pH . Vesicles of two opposite orientations were prepared from erythrocytes with fused G-protein . Right-side-out and inside-out vesicles expose the exoplasmic and the cytoplasmic domains of the G-protein on their surfaces respectively . In immuno-isolation experiments antibodies against each of the domains of the G-protein were used . Vesicles were presented to an immunoadsorbent (ImAd) consisting of a solid support with appropriate antibodies bound to its surface . Two commonly used immunoadsorbents prepared from either polyacrylamide beads or fixed Staphylococcus aureus cells were compared and found to have identical immuno-isolation efficiencies . It was possible to control and quantitate the amount of implanted antigen . Therefore, we were able to show that the critical antigen density required for immuno-isolation is 50 G molecules/micron2 plasma membrane surface area for both types of vesicle/antibody couples . This analysis showed that vesicles presenting either the cytoplasmic or the exoplasmic domain of the G-protein are immuno-isolated with the same efficiency. Proc Natl Acad Sci U S A, 1985 Sep, 82(17), 5666 - 70 Amino acid sequence of the UP1 calf thymus helix-destabilizing protein and its homology to an analogous protein from mouse myeloma; Williams KR et al.; A complete amino acid sequence has been determined for the UP1 single-stranded DNA binding protein from calf thymus that was first described by G . Herrick and B . M . Alberts {(1976) J . Biol . Chem . 251, 2124-2132} . Peptides required to establish the UP1 sequence were isolated by reversed-phase HPLC of digests produced by endoproteinase Lys-C, trypsin, chymotrypsin, Staphylococcus aureus V8 protease, and cyanogen bromide cleavage of UP1 . The purified peptides were coupled to aminopolystyrene prior to solid-phase sequencing . UP1 contains 195 amino acids and has a molecular weight of 22,162 . UP1 has a blocked NH2 terminus and contains a single NG,NG-dimethylarginine residue near its COOH terminus . Gas-phase sequencing of tryptic peptides derived from an analogous protein from mouse myeloma cells {Planck, S . R . & Wilson, S . H . (1980) J . Biol . Chem . 255, 11547-11556} revealed that this mouse helix-destabilizing protein shares a high degree of sequence homology with UP1 . Of the 59 amino acids in the mouse protein that have so far been found to be homologous with UP1, 48 correspond exactly to sequences found in UP1 . Most of the 11 differences that have been found between the sequences of these two proteins are conservative in nature, involving primarily the interchange of chemically similar amino acids . One 9-residue mouse sequence that is not obviously homologous to UP1 may be a result of the larger size of the mouse myeloma protein as compared to UP1 . Since none of the UP1 or mouse myeloma helix-destabilizing protein sequence appears to be homologous to that of any previously sequenced protein, we presume that these two proteins represent a distinct class of single-stranded nucleic acid binding proteins that probably play a role in metabolism of single-stranded RNA or DNA in vivo. J Hosp Infect, 1985 Sep, 6(3), 285 - 92 Control of a hospital outbreak of methicillin-resistant Staphylococcus aureus infections: value of an isolation unit; Shanson DC et al.; An outbreak of methicillin- and gentamicin-resistant Staphylococcus aureus infections started in a university hospital after a patient, who was not known to be colonized, was admitted . During a 3-month period 15 surgical or geriatric patients and five staff were found to be infected or colonized by the epidemic strain in five surgical/orthopaedic wards, a geriatric ward, the intensive care unit and the isolation unit . Difficulties in controlling the outbreak arose when two patients who initially had negative bacteriological screening results were returned to general wards . Both patients were subsequently shown to be colonized and caused outbreaks which led to the further closure of two general wards . There was strong circumstantial evidence to suggest that physiotherapy staff were involved with the spread of the epidemic strain . Control of the outbreak was achieved by more strict isolation of 'negative' patient contacts as well as colonized/infected patients and increasing the level of staffing on the separate isolation unit. Schweiz Med Wochenschr, 1985 Aug 31, 115(35), 1196 - 9 {Frequency of toxic-shock-syndrome-toxin producing strains of Staphylococcus aureus in the Basel region}; Vischer WA et al.; An attempt was made to explain the fact that toxic shock syndrome (TSS) is far less often observed among Swiss women than in the USA . To this end, 353 patients were examined for vaginal infection with Staphylococcus aureus, and 131 strains of Staphylococcus aureus of various origins tested for the formation of "toxic shock syndrome toxin one" (TSST-1) . In addition, the patients were questioned about the use of tampons for menstrual hygiene . Taking all age groups into consideration, 2.3% of the 353 patients, and 4.8% of those younger than 27 years, were Staphylococcus aureus carriers . 19.8% of the tested strains of Staphylococcus aureus produced TSST-1 . Since these figures are comparable with the US statistics, they afford no explanation for the difference in the incidence of TSS . On the other hand, it seems very probable that the much less common use of tampons, especially the highly absorbent variety, could be responsible . It is also possible that TSS is less often recognized and reported in Switzerland. Biochem Biophys Res Commun, 1985 Aug 30, 131(1), 484 - 91 Synergism between diacylglycerols and calcium ionophore in the induction of human B cell proliferation mimics the inositol lipid polyphosphate breakdown signals induced by crosslinking surface immunoglobulin; Guy GR et al.; Resting human tonsillar B cells were stimulated to divide by heat killed Staphylococcus aureus Cowan strain 1 which was shown to induce hydrolysis of phosphatidylinositol 4, 5-bisphosphate known to give rise to diacylglycerol and an increase in cytosolic free calcium . Addition of the diacylglycerols, 1-oleoyl-2 acetyl glycerol or sn-1, 2-dioctanoylglycerol, together with the calcium ionophore ionomycin to B cell cultures induced marked cell proliferation whereas these agents were ineffective when used alone . Both diacylglycerols were shown to compete with {3H} phorbol 12,13 dibutyrate in binding to protein kinase C . These data support the hypothesis that synergism between cytosolic calcium and endogenous diacylglycerol, which activates protein kinase C, is involved in signal transduction in the proliferation of human B cells. J Biol Chem, 1985 Aug 25, 260(18), 10185 - 91 Tyrosine 264 in the recA protein from Escherichia coli is the site of modification by the photoaffinity label 8-azidoadenosine 5'-triphosphate; Knight KL et al.; The photoaffinity label 8-azidoadenosine 5'-triphosphate (N3-ATP) was used to covalently modify the recA protein from Escherichia coli within its ATP-binding site . We have previously demonstrated that N3-ATP modification of recA protein is specific for the ATP-binding site and have isolated a unique tryptic peptide (T31), spanning residues 257-280, that contains the exclusive site of attachment of this ATP analog (Knight, K . L., and McEntee, K . (1985) J . Biol . Chem . 260, 867-872) . We performed a secondary proteolytic digestion of the {alpha-32P}N3-ATP-labeled T31 peptide using Staphylococcus aureus V8 protease and purified the resulting peptide fragments by high-pressure liquid chromatography (HPLC) . Based on a comparison of the amino acid compositions of all purified fragments and sequence analysis of one labeled fragment we determined that Tyr-264 is the exclusive site of N3-ATP attachment in recA protein . Photoaffinity labeling of recA protein was also performed in the presence of single-stranded DNA . Following trypsin treatment and separation of peptides by HPLC we showed that tryptic peptide T31 contained the exclusive site of N3-ATP attachment . A secondary proteolytic digestion was performed on both {alpha-32P}N3ATP-modified T31 and unmodified T31 using alpha-chymotrypsin . Comparison of the HPLC profiles and amino acid compositions of the resulting fragments was consistent with Tyr-264 as the exclusive site of N3-ATP attachment to recA protein. J Biol Chem, 1985 Aug 25, 260(18), 10177 - 84 Affinity labeling of a tyrosine residue in the ATP binding site of the recA protein from Escherichia coli with 5'-p-fluorosulfonylbenzoyladenosine; Knight KL et al.; We have covalently modified the recA protein from Escherichia coli with the adenine nucleotide analog 5'-p-fluorosulfonylbenzoyladenosine (5'-FSBA) . The rate at which the protein is modified shows a sigmoidal dependence on the concentration of 5'-FSBA suggesting that binding of the analog is characterized by positive cooperativity . Covalent modification of the protein results in irreversible inactivation of its single-stranded DNA-dependent ATPase activity such that 100% inactivation is achieved when 25% of the enzyme monomers have been modified . Attachment of 5'-FSBA is specific for the ATP-binding site of recA protein as judged by the following criteria: (i) attachment of the affinity label to the protein appears to saturate at 1 mol of 5'-FSBA/mol of protein; (ii) binding of 5'-FSBA to recA protein is inhibited by ATP and competitive inhibitors of its ATP hydrolytic activity, e.g . adenosine-5'-O-(thiotriphosphate), ADP, UTP, and GTP, but not by adenosine; (iii) attachment of 5'-FSBA to the protein occurs at a single site as determined by high pressure liquid chromatography peptide separation . Following trypsin digestion of recA protein that had been covalently modified with {3H}5'-FSBA we isolated a single labeled peptide (T31) containing the exclusive site of 5'-FSBA attachment . A secondary proteolytic digestion was performed on both 5'-FSBA modified T31 and unmodified T31 using Staphylococcus aureus V8 protease, and by comparison of the amino acid compositions of the resulting peptides we identified Tyr-264 as the exclusive site of 5'-FSBA attachment in recA protein. Br Med J (Clin Res Ed), 1985 Aug 24, 291(6494), 501 - 4 Impaired phagocytic activity of neutrophils in patients receiving haemodialysis: the critical role of iron overload; Waterlot Y et al.; The metabolic burst (as measured by the spontaneous and stimulated nitroblue tetrazolium tests), the phagocytosis of heat inactivated bakers' yeast and of Staphylococcus aureus, the killing of Staph aureus, and the myeloperoxidase activity of polymorphonuclear neutrophils were studied in 11 patients receiving maintenance haemodialysis . Of these patients, six were polytransfused and had high serum ferritin concentrations (mean 5940 (SD 2925) micrograms/l; group 1), and five had normal serum ferritin values (mean 171 (116) micrograms/l; group 2) . Patients in group 1 had a history of more infectious episodes (0.167 v 0.025 per patient per month) and significantly more genitourinary infections (p = 0.015) than those in group 2 . Phagocytosis and myeloperoxidase activity were severely reduced in group 1 but normal in group 2 . Percentages of neutrophils ingesting one or more particles together with the index of phagocytosis in patients' serum were inversely correlated with serum ferritin concentrations . Four patients in group 1 were treated with desferrioxamine, and after six to 18 weeks of treatment phagocytosis and myeloperoxidase activity had returned to normal in three of them . These data suggest that in patients receiving haemodialysis iron overload due to multiple transfusions plays an important part in the mechanisms underlying the susceptibility to bacterial infections, mediated at least partially through impaired neutrophil function. J Biol Chem, 1985 Aug 15, 260(17), 9567 - 73 The primary structure of thioredoxin from the filamentous cyanobacterium Anabaena sp . 7119; Gleason FK et al.; Thioredoxin from the cyanobacterium Anabaena 7119 serves as electron donor to ribonucleotide reductase and as a protein disulfide reductase . This small, heat-stable protein was found to have structural and functional similarities to thioredoxins from both bacterial and mammalian sources . We here report the complete primary structure of Anabaena thioredoxin . The structure was determined by analysis of peptides obtained after cleavage with cyanogen bromide, Staphylococcus aureus protease, and trypsin . The protein consists of 106 residues with the following amino acid sequence: Ser-Ala-Ala-Ala-Gln-Val-Thr-Asp- Ser-Thr-Phe-Lys-Gln-Glu-Val-Leu-Asp-Ser-Asp-Val-Pro-Val-leu-Val-Asp-Phe- Trp-Ala-Pro-Trp-Cys-Gly-Pro-Cys-Arg-Met-Val-Ala-Pro-Val-Val-Asp-Glu- Ile-Ala-Gln-Gln-Tyr-Glu-Gly-Lys-Ile-Lys-Val-Val-Lys-Val-Asn-Thr-Asp- Glu-Asn-Pro-Gln-Val-Ala-Ser-Gln-Tyr-Gly-Ile-Arg-Ser-Ile-Pro-Thr-Leu- Met-Ile-Phe-Lys-Gly-Gly-Gln-Lys-Val-Asp-Met-Val-Val-Gly-Ala-Val-Pro- Lys-Thr-Thr-Leu-Ser-Gln-Thr-Leu-Glu-Lys-His-Leu . The sequence of Anabaena thioredoxin shows a definite homology to the protein from Escherichia coli, with 49% residue identities occurring in the proteins when aligned at the active site disulfide. Anal Biochem, 1985 Aug 15, 149(1), 153 - 62 Isolation, characterization, and amino-terminal amino acid sequence analysis of human neutrophil elastase from normal donors; Heck LW et al.; Human neutrophil elastase from normal donors has been purified using an isolation procedure which included sequential sodium chloride extraction, Aprotinin-Sepharose affinity chromatography, CM-cellulose ion-exchange chromatography, and AcA44 gel filtration chromatography . The inclusion of this last purification step was crucial for separating inactive lower molecular weight species from the active forms of neutrophil elastase and resulted in a higher specific activity of the final preparation . Sodium dodecyl sulfate-polyacrylamide gradient gel electrophoresis of the reduced purified protein demonstrated three polypeptides of Mr 31,000, 28,000, and 27,500 . Four polypeptides were resolved on acid gel electrophoresis; each of the four possessed amidolytic activity . Furthermore, peptide analysis of Staphylococcus aureus V8 protease digests indicated that these polypeptides are structurally related to each other and represent microheterogeneity of the purified protein . The apparent isoelectric points of these four forms as determined by two-dimensional electrophoresis range from 6.1 to 6.7 . By utilizing microsequencing techniques, the first 40 residues of neutrophil elastase have been determined and compared with the reported sequence of elastase isolated from leukemic myeloid cells . In addition, a high degree of homology was found within the amino-terminal regions of neutrophil elastase and the serine proteinases porcine elastase, bovine chymotrypsin, human factor D, and the beta chain of plasmin. Aust J Exp Biol Med Sci, 1985 Aug, 63 ( Pt 4), 463 - 72 A plasmid for diffusible pigment production in Staphylococcus aureus; Townsend DE et al.; Production of diffusible pigment in a strain of Staphylococcus aureus (WG260) has been associated with a plasmid of c.26 megadaltons which can be co-transduced with inducible erythromycin and spectinomycin resistance . However, the diffusible pigment plasmid can be lost from the transductants without loss of inducible erythromycin and spectinomycin resistance . The evidence suggests that the resistance determinants are located on an element similar or identical to transposon Tn554, and, during transduction of the diffusible pigment plasmid, a copy of the resistance determinants always inserts into the chromosome of the transductant . This is the first report of such an element occurring on a plasmid in a clinical isolate of S . aureus . To date, the diffusible pigment plasmid has been isolated only as the open-circular conformation of plasmid DNA and is probably a highly relaxable plasmid . The production of the orange, diffusible pigment was shown to be independent of colony pigmentation. Acta Orthop Scand, 1985 Aug, 56(4), 327 - 9 Antibiotic prophylaxis in lower limb amputation; Moller BN et al.; We have prospectively studied the effect of 1-day prophylactic antibiotic therapy in lower-limb amputation for ischaemia . Twenty-seven patients were treated with Meticillin 1 g X 4 intravenously on the day of operation; 23 control patients did not receive any antibiotics . Eight patients in the control group had postoperative wound infections compared to none in the Meticillin group . Seven patients were re-amputated because of infection . Preoperatively, Staphylococcus aureus was isolated in 5/8 of the patients in the Meticillin and 6/8 in the control group . In the postoperatively infected stumps, S . aureus occurred in 6/8 of the patients in the control group, and one patient developed gas gangrene. Liver, 1985 Aug, 5(4), 205 - 11 Detection of two forms of HBeAg (free-and IgG-bound HBeAg) in patients with HBe antigenemia using staphylococcus bearing protein A; Sagnelli E et al.; Protein A-bearing Staphylococcus aureus organisms (STA) were used to separate free HBeAg from IgG-bound HBeAg . Free HBeAg was detected in the supernate while IgG-bound HBeAg could be liberated from the pellets using MgCl2 or a glycine buffer . HBeAg was determined by radioimmunoassay and the results expressed as patient's cpm/normal control's cpm ratio (S/N ratio) . This ratio was demonstrated to be proportionate to the antigen concentration and used as a titer of HBeAg . Sera of 40 HBsAg-negative healthy volunteers and 82 HBeAg-positive patients who were either asymptomatic HBsAg carriers or had various diseases including chronic persistent hepatitis (CPH), chronic active hepatitis (CAH), and renal disease undergoing hemodialysis, were tested for free HBeAg and IgG-bound HBeAg . Patients with CAH from two different countries were compared . Free HBeAg was detected in all patients but one, IgG-bound HBeAg was detected with similar prevalences (from 56% to 67%) in HBsAg asymptomatic carriers, hemodialysis patients, CPH and Italian CAH patients . In contrast, all CAH patients from New York, who had frequently been exposed to HBV infection, had detectable levels of IgG-bound HBeAg, with the highest S/N ratios observed in the study, and frequently showed an unfavorable outcome . In AVH due to HBV and delta agent co-infection, IgG-bound HBeAg was detected in two of four patients only in the initial stage of the disease . The data reported indicate that a separate determination of free- and IgG-bound HBeAg may have clinical value. Acta Pathol Microbiol Immunol Scand {B}, 1985 Aug, 93(4), 307 - 13 Antibody response against whole Staphylococcus aureus in patients with staphylococcal septicaemia and endocarditis investigated by ELISA; Jarlov JO et al.; A whole cell Staphylococcus aureus enzyme-linked immunosorbent assay (ELISA) for the detection of IgG antibodies against S . aureus has been developed . To avoid non-specific binding of IgG to protein A, the protein A-poor strains of S . aureus, E 1369 and Wood 46, were used as antigens . One-hundred and eighty serum samples from 120 patients with S . aureus endocarditis, non S . aureus endocarditis, S . aureus septicaemia and non S . aureus septicaemia were tested together with sera from 155 healthy controls . The sensitivity was similar for the E 1369 ELISA and the Wood 46 ELISA and positive test values were detected in 84.2% of patients with S . aureus endocarditis and 41.2% of patients with complicated S . aureus septicaemia . No distinction could be found between complicated and uncomplicated S . aureus septicaemia . The E 1369 ELISA was more specific showing cross reactions with sera from patients infected with other bacteria than S . aureus in 3.6% . Furthermore, the reproducibility was better for the E 1369 ELISA with a coefficient of variation at 0.054 . The absence of need for purified antigens makes the whole S . aureus ELISA easy, rapid and cheap . Therefore, we suggest the whole S . aureus ELISA as a good alternative to previously reported assays using purified cell wall antigens. Curr Eye Res, 1985 Aug, 4(8), 905 - 12 Bovine lens calmodulin . Isolation, partial characterization and calcium-independent binding to lens membrane proteins; van den Eijnden-van Raaij AJ et al.; Calmodulin has been isolated from calf lens fiber cells . Like other vertebrate calmodulins lens calmodulin shows a calcium-dependent mobility shift on SDS-polyacrylamide gels and forms immune complexes with antiserum, raised against vertebrate calmodulin . Via the gel overlay technique radioiodinated calmodulin from lens or bovine brain was found to bind to the main intrinsic protein (MIP) and the 17.5 kDa protein of lens fiber membranes in a calcium-independent manner . After proteolytic digestion of lens fiber membranes with trypsin or Staphylococcus aureus V8 protease the calmodulin-binding activity of MIP is retained . This result indicates that the small polypeptide fragment of MIP, which is accessible to proteolytic attack, apparently is not the attachment point for calmodulin . Two additional calmodulin-binding proteins (MW 14 kDa and 16.5 kDa) are observed in junction-enriched fiber membrane fractions . These junction-specific proteins are bound to the membrane via calcium . In addition to MIP and the 17.5 kDa protein they are possibly involved in the calcium-dependent regulation of lens fiber junctions . The 14 and 16.5 kDa proteins are also present in epithelial membranes, prepared from freshly obtained calf lens epithelia . Whereas in the latter membranes the two proteins form part of the four 14-17 kDa major protein components, these proteins are absent in membranes from cultured lens epithelial cells . The epithelial 14 kDa and 16.5 kDa proteins thus appear to be junction-specific . The capacity of the latter proteins to bind calmodulin in the presence and absence of calcium indicates that these junction-specific proteins are very similar, if not identical, to the corresponding fiber junctional proteins.(ABSTRACT TRUNCATED AT 250 WORDS) Curr Eye Res, 1985 Aug, 4(8), 821 - 9 Efficacy of topical chloramphenicol and tobramycin ophthalmic solutions in preventing severe Staphylococcus aureus keratitis in rabbits; Edwards JG et al.; Various marketed chloramphenicol ophthalmic solutions were compared and various dilutions of Tobrex Ophthalmic Solution were tested for effectiveness in a Staphylococcus aureus rabbit keratitis model . Anesthetized rabbits were each infected intracorneally with 10(4) Staphylococcus aureus ATCC 29737 cells . Treatment groups consisted of five or six rabbits (10 or 12 eyes) per group . One group of rabbits was infected but not treated (Positive Control Group) . Topical dosing of commercially available ophthalmic solutions was accomplished by depositing 0.1 mL of a color-coded test solution into the lower cul-de-sac of each eye . Dosing begin one hour after the mid-infection time and continued for a total of nine hourly treatments . Twenty-four hours after infection the rabbit eyes were graded (masked) using standard slit-lamp scoring procedures . The slit-lamp scores for five of eight ocular parameters were used to calculate an eye score value for each rabbit eye . The five ocular parameters were selected, based on previous Stepwise Discriminant Computer Analysis of over 300 rabbit eyes infected with Staphylococcus aureus ATCC 29737 and treated with various antibiotics . The eye score values for each group were averaged and the treatments were compared for significant differences in efficacy using the nonparametric, Wilcoxon-Mann-Whitney Rank Sum Test.(ABSTRACT TRUNCATED AT 250 WORDS) J Infect Dis, 1985 Aug, 152(2), 286 - 92 A new serological assay for Staphylococcus aureus infections: detection of IgG antibodies to S . aureus lipase with an enzyme-linked immunosorbent assay; Christensson B et al.; Purified Staphylococcus aureus lipase was used as antigen in an enzyme-linked immunosorbent assay (ELISA) that detected IgG antibodies in 169 patients with infections due to S . aureus, in 122 patients with infections not due to S . aureus, and in 167 healthy controls . Eighty-eight percent (21 of 24) of the patients with endocarditis due to S . aureus showed a positive level of antibody to lipase or a significant change in antibody titer during the first month, as did 89% (17 of 19) and 28% (5 of 18) of the patients with complicated and uncomplicated septicemia due to S . aureus, respectively . The specificity for S . aureus infections was high; only one patient in the non-S . aureus endocarditis and septicemia groups showed a significant rise in antibody titer, and this rise did not reach a positive antibody level . Patients with recurrent furunculosis or chronic osteomyelitis due to S . aureus responded in only 15% and 23% of cases, respectively . We suggest that the antibody-to-lipase ELISA could