J Infect Dis, 1987 Feb, 155(2), 242 - 6 Capsular polysaccharide antigenemia in rats with experimental endocarditis due to Staphylococcus aureus; Arbeit RD et al.; In vivo expression of the type 8 capsular polysaccharide (CP) of Staphylococcus aureus was studied in the rat model of aortic valve endocarditis . An enzyme-linked immunosorbent assay (ELISA) with monoclonal antibodies to type 8 CP was sensitive to 0.16 ng/ml in serum . All (23 of 23) animals infected with the prototype type 8 strain Becker had CP detected in one or more serum samples . Peak CP levels ranged from 0.16 to 240 ng/ml (median, 25 ng/ml) . Among the 14 rats alive on day 2, antigenemia and bacteremia were significantly correlated (r = .71, P less than .005) . CP was also present in serum of rats infected with three additional type 8 bacteremic isolates . All serum samples from animals infected with the type 5 prototype strain were negative in the type 8 ELISA . These studies document that the type 8 CP of S . aureus, previously identified only in vitro, is produced and released during in vivo infection and can be detected in serum of infected animals. J Neurochem, 1987 Feb, 48(2), 644 - 52 Expression of a 48-kilodalton growth-associated protein in the goldfish retina; Perrone-Bizzozero NI et al.; One of the most striking molecular correlates of optic nerve regeneration in the goldfish is the increased labeling of a 48 kilodalton (kD) acidic protein that is conveyed to the developing nerve endings from the retina by rapid axonal transport . The present study examined the biosynthesis and molecular characteristics of this protein . Retinas derived either from intact controls or from goldfish undergoing optic nerve regeneration (10-14 days postcrush) were pulse-labeled with {3H}proline or {35S}methionine, followed by subcellular fractionation and analysis of protein synthesis patterns by two-dimensional gel electrophoresis and fluorography . Synthesis of the 48-kD acidic protein (termed here GAP-48) was detected only in retinas that were undergoing axonal regeneration . Pulse-chase labeling experiments demonstrated that the protein undergoes a post-translational modification that requires 15-20 min . This processing could be selectively blocked by tunicamycin, an inhibitor of protein N-glycosylation . The protein was also found to incorporate low levels of phosphate in vitro . Thus, the differential appearance of GAP-48 in regenerating axons might be regulated either at the level of gene expression or by selective posttranslational processing in retinal ganglion cells . By the criteria of molecular weight, isoelectric point, anomalous migration properties on sodium dodecyl sulfate-polyacrylamide gels, phosphorylation, subcellular distribution, and the pattern of digestion products generated by Staphylococcus aureus V8 protease, GAP-48 appears to be equivalent to the B-50 (F-1) phosphoprotein of the mammalian brain. J Gen Microbiol, 1987 Feb, 133 ( Pt 2), 431 - 8 Immunological characterization of an exopolysaccharide from the Staphylococcus aureus strain Smith diffuse; West TE et al.; Exopolysaccharides (EXPs) of Staphylococcus aureus are associated with virulence in animal models . An EXP from the S . aureus strain Smith diffuse was previously detected in 64.3% of S . aureus clinical isolates . EXP was isolated from culture supernatants of this strain after DNAase, RNAase, phosphodiesterase I and lysostaphin treatment, and was further purified by cation-exchange and molecular-sieve chromatography . Isoelectric focusing revealed a pI of 3.6 for the EXP while the pI of teichoic acid was less than 2.7 . Crossed immunoelectrophoresis with homologous Smith diffuse antisera indicated that the EXP contained two immunological components . A major precipitin line persisted after the antisera had been absorbed with the non-EXP-producing variant strain, Smith compact, while the second component was removed . Tandem immunoelectrophoresis also demonstrated that the EXP was distinct from teichoic acid . The EXP contained 2-amino-2-deoxyglucuronic acid, glucose, mannose and galactose . No fatty acids or nucleic acids were present and total protein content was less than 2% . Teichoic acid could not be demonstrated in the EXP, thus further substantiating the immunological studies . S . aureus EXP isolated by the present method can be used for further serological and virulence studies. Jpn J Antibiot, 1987 Feb, 40(2), 349 - 56 {Antibacterial activities of arbekacin, a new aminoglycoside antibiotic, against methicillin-cephem-resistant Staphylococcus aureus}; Watanabe T et al.; The in vitro and in vivo antibacterial activities of a new aminoglycoside antibiotic, arbekacin (HBK), against methicillin-cephem-resistant Staphylococcus aureus (MRSA) were compared with those of gentamicin (GM), netilmicin (NTL) and amikacin (AMK) . The results obtained were summarized as follows: Compared to other aminoglycoside antibiotics, HBK had the highest antibacterial activities against clinically isolated MRSA (46 strains) . Therapeutic effects of HBK against experimental systemic infections with MRSA in mice, were superior to those of GM, NTL and AMK . The ED50's of GM, NTL and AMK were more than 2 mg/mouse . Therapeutic effects of HBK against experimental subcutaneous infections with MRSA in mice were also superior to those of GM, NTL and AMK. Am J Infect Control, 1987 Feb, 15(1), 1 - 6 Staphylococcus aureus bacteremia caused by infected intravenous catheters; Mylotte JM et al.; A prospective study of all cases of Staphylococcus aureus bacteremia (SAB) has been ongoing at our institution since April 1983 . Data collected as of December 1984 reveal that of 79 episodes of SAB identified, 28 (35%) were caused by infected intravenous catheters; 14 of these were peripheral catheters and 14 were subclavian catheters . Endocarditis or metastatic infection was not recognized; two patients, one with a prosthetic heart valve and the other with recurrent SAB, did not meet the criteria for these complications but the clinical circumstances were strongly suggestive . Six of the 28 patients (21%) died of their infection, and local complications (inflammation or purulent drainage at the insertion site) were common . Only four of 22 patients who recovered were administered antibiotics for greater than 14 days . We conclude that patients with SAB caused by an infected intravenous catheter have a low risk of endocarditis or metastatic infection when the infected catheter is promptly removed and no risk factors (valvular heart disease or a prosthetic valve) are present that favor development of endocarditis . The duration of antimicrobial therapy need not exceed 2 weeks in this situation . The mortality related to SAB, however, was high and emphasizes the need to develop methods to prevent this infection. J Clin Microbiol, 1987 Feb, 25(2), 412 - 5 Detection of intrinsically resistant (heteroresistant) Staphylococcus aureus with the Sceptor and AutoMicrobic systems; Hansen SL et al.; Modified procedures for the Sceptor Gram-Positive MIC Panel and the Vitek AutoMicrobic System GPS-M Card were evaluated for their ability to detect methicillin-resistant (heteroresistant) Staphylococcus aureus . A total of 398 clinical isolates (including 222 methicillin-resistant S . aureus) obtained from 10 hospitals were tested . Both systems had 2% NaCl in the oxacillin wells . Sceptor MIC panels were inoculated with an organism suspension prepared from an 18- to 24-h blood agar plate and were inoculated for a full 24 h at 35 degrees C before MICs were read . All methicillin-resistant S . aureus isolates were detected as resistant to oxacillin at greater than or equal to 8 micrograms/ml by the Sceptor method and at greater than 2 micrograms/ml by the Vitek method . All 176 oxacillin-susceptible, methicillin-susceptible S . aureus isolates were correctly distinguished from methicillin-resistant S . aureus isolates by Sceptor . However, with the Vitek system 29 methicillin-susceptible S . aureus isolates tested as falsely resistant to oxacillin and four isolates tested as falsely resistant to vancomycin . The modified testing procedure with the Sceptor system can be used reliably for accurate susceptibility testing of methicillin-resistant and methicillin-susceptible S . aureus . The Vitek GPS-M card does not accurately discriminate between methicillin-resistant and methicillin-susceptible S . aureus with an oxacillin breakpoint of greater than 2 micrograms/ml. Poult Sci, 1987 Feb, 66(2), 203 - 8 Major histocompatibility (B) complex-associated differences in the delayed wattle reaction to staphylococcal antigen; Cotter PF et al.; The influence of the major histocompatibility (B) complex on the delayed wattle reaction (DWR) to Staphylococcus aureus was studied in 109 segregants (B2/B2, B2/B5, and B5/B5) of a fourth generation cross between inbred Regional Poultry Research Laboratory lines 6(1) and 15(1) . Chickens were sensitized at 6 weeks of age with S . aureus antigen . One week later, DWR was evaluated by injecting the right wattle with S . aureus antigen . Thickness measurements were taken 4, 24, 48, 72, and 96 hr after injection . A quadratic equation model was used for each bird to calculate maximum wattle thickness, hour of maximum response, and rate of response development and decline . In males, the maximum wattle thickness in response to S . aureus antigen was significantly greater in B2/B5 heterozygotes (1.77 +/- .07 mm) than in either homozygote, B2/B2 (1.36 +/- .13 mm) or B5/B5 (1.39 +/- .08 mm) . Heterozygous males reached maximum response sooner and recovered more quickly than homozygous males but these differences were not statistically significant . In females, response developed later than in males but no B complex effect was detected in either rate of development or maximum response. Acta Orthop Scand, 1987 Feb, 58(1), 14 - 9 Rabbit model of septic arthritis; Riegels-Nielson P et al.; Eighty-five rabbits were injected in one knee with Staphylococcus aureus in order to study the time-related changes in untreated septic arthritis up to 3 months . In the synovial membrane a severe release of lysosomal enzymes was observed . The activity was mainly located in and around lining cells and leucocytes in the pannus demonstrating increasing destructive characteristics . This resulted in marginal erosion and undermining of the cartilage border visible from Day 5 continuing gradually to total joint destruction after 5 weeks . The glycosaminoglycan depletion was observed at the surface of the cartilage at Day 2 and was total after 2 weeks . Our infection model should permit comparison of different therapeutic measures. Geburtshilfe Frauenheilkd, 1987 Feb, 47(2), 104 - 6 {Toxic shock syndrome toxin-1 producing strains of Staphylococcus aureus in vaginal smears}; Dickgiesser N et al.; 21 Staphylococcus aureus strains were isolated from 391 vaginal swabs . This corresponds to a frequency of 5.3% . In females using tampons this frequency was 7%, in users of napkins 2.4% . 2 out of these 21 S . aureus strains produced toxic shock syndrome toxin-1 (TSST-1) . Both strains originated from women using tampons . Therefore, 5 out of 1000 women must be expected to carry TSST-1-producing S . aureus strains within their vagina. Antimicrob Agents Chemother, 1987 Feb, 31(2), 259 - 63 Comparative evaluation of A-56619, A-56620, and nafcillin in the treatment of experimental Staphylococcus aureus osteomyelitis; Mader JT et al.; A rabbit model for Staphylococcus aureus osteomyelitis was used to compare the results of treatment with A-56619 and A-56620, two new aryl-fluoroquinolones, and nafcillin . A-56619 (15 mg/kg) and A-56620 (20 mg/kg), both used for 28 days of treatment, were injected subcutaneously every 12 h, and nafcillin (40 mg/kg) was injected every 6 h . After treatment, S . aureus was found on bone marrow cultures from 19 of 20 control rabbits, 6 of 20 treated with A-56619, 14 of 20 treated with A-56620, and 8 of 20 treated with nafcillin . Drug concentrations in serum and uninfected and infected bone were measured 1 h after A-56619 and A-56620 injection and 30 min after nafcillin injection in a group of rabbits that had been infected for 3 to 4 weeks . The concentrations in infected bone were similar for all three drugs and were significantly higher than in uninfected bone . The results of this study showed that A-56619 had a high rate of eradication of S . aureus from infected bone and compared favorably to nafcillin. Vet Immunol Immunopathol, 1987 Feb, 14(2), 145 - 56 Use of enzyme-linked immunosorbent assay to measure bovine milk and serum antibodies to alpha toxin, beta toxin, and capsular antigens of Staphylococcus aureus; Loeffler DA et al.; An enzyme-linked immunosorbent assay (ELISA) procedure was used to quantitate milk and serum antibodies (IgG) to Staphylococcus aureus alpha and beta toxins, and S . aureus 2-8 and Smith diffuse strain capsular antigens . Milk samples were collected on two occasions . A comparison was made between levels of milk antibodies specific for the two toxins and capsular antigens for 41 cows that were infected with S . aureus on both sampling dates, and 18 cows not S . aureus-infected on either date . Staphylococcus aureus-infected cows were grouped according to somatic cell counts . All groups of infected cows, regardless of somatic cell counts, had significantly higher milk antibody levels to alpha and beta toxins than did the non-infected cows (P less than .002) . Serum samples taken for 13 infected and 4 non-infected cows also indicated that significant elevations in anti-alpha toxin and anti-beta toxin IgG were present in S . aureus-infected cows, compared to non-infected cows . A similar immune response was not seen to capsular antigens, however . No significant differences were present between the two groups of cows for either milk or serum antibodies to Smith diffuse strain capsular antigens . Milk antibodies to 2-8 capsule were significantly elevated only in infected cows with somatic cell counts greater than 10(6)/ml, compared to non-infected cows; no differences were present for serum antibodies to 2-8 capsule between infected and non-infected cows . These results indicate that significant increases in milk (and possibly serum) antibodies to alpha and beta toxins are present in cows with chronic staphylococcal mastitis, apparently resulting from a systemic immune response to these toxins . There does not appear to be a similar immune response to capsular antigens. Scott Med J, 1987 Feb, 32(1), 28 - 9 Toxic shock syndrome (TSS) in children; Galea P et al.; Toxic shock syndrome is uncommon in the prepubertal age group . Two children presented with pyrexia, macular erythroderma, vomiting, hypotension and rapid deterioration of consciousness . One child had severe neurological involvement . The diagnosis of toxic shock syndrome was established in both cases by the exclusion of other causes and by culturing staphylococcus aureus . We postulate that the neurological manifestations were caused by a direct neurotoxic action of the staphylococcal-produced toxin . Both children made a complete recovery. Infect Immun, 1987 Feb, 55(2), 428 - 32 Expression of staphylococcal enterotoxin C1 in Escherichia coli; Bohach GA et al.; The structural gene encoding staphylococcal enterotoxin C1 was cloned into Escherichia coli and localized on a 1.5-kilobase HindIII-ClaI DNA fragment by subcloning . The toxin was partially purified from E . coli clones and shown to be immunologically identical to enterotoxin C1 from Staphylococcus aureus . The cloned toxin also had the same molecular weight (26,000) and charge heterogeneity as staphylococcus-derived enterotoxin . Toxins from both sources were equally biologically active. J Clin Endocrinol Metab, 1987 Feb, 64(2), 246 - 54 Human anti-estrogen receptor antibodies: assay, characterization, and age- and sex-related differences; Mudarris A et al.; A binding assay was developed to measure the estrogen receptor (ER)-binding activity in serum . The method employed immunoglobulin G (IgG) adsorption with protein A-bearing Staphylococcus aureus cells before incubation with tritium-labeled ER . Specificity studies employing chromatographic and electrophoretic analyses suggested that an IgG antibody (Anti-ER) was responsible for the serum activity . Anti-ER was found in human, rat, and mouse serum and exhibited species cross-reactivity . The antibody recognized both the 8S and 5S forms of the ER . Anti-ER was measured in 262 individuals ranging in age from 1-85 yr . The antibody was detected in all serum samples examined, suggesting its natural occurrence . When the study group was divided into three arbitrary age groups (young, 1-13 yr; middle age, 13-51 yr; older, greater than 51 yr), significant differences in levels of antibody were found, with highest levels in the young, followed by the elderly, and lowest levels in the middle years . An examination of sex- as well as age-related differences in the population revealed a striking sex difference . Thus, the young male population had a lower level of antibody than the corresponding female population, and these levels in males declined throughout life to reach their lowest point in old age, whereas the high levels in young females declined in middle years and then increased significantly in the elderly . We postulate that the antibody is a subgroup of IgG of multifactorial etiology . The idiotypic network theory offers an explanation for the occurrence of these antibodies in normal serum, whereas an autoimmune mechanism could account for the secondary rise of anti-ER in the aging female population. Exp Hematol, 1987 Feb, 15(2), 115 - 8 Human stem cell colony-stimulating activity (CFU-GEMMCSA) in medium conditioned by leukemic B-lymphocytes; Merchav S et al.; Media conditioned by B-lymphocytes of patients with chronic lymphocytic leukemia (B-CLL) induced the formation of multilineage colonies (CFU-GEMM) in cultures of human target marrow cells . Maximum levels of stem cell colony-stimulating activity (CFU-GEMMCSA) were detected by day 3 of leukemic B-cell cultures, remaining constant thereafter . Stimulation of leukemic B-lymphocytes with B-cell mitogens such as Cowan strain Staphylococcus aureus (SAC) and anti-mu enhanced the production of CFU-GEMMCSA by these cells . 3H-thymidine uptake in response to these mitogens was detected only in SAC-stimulated cultures, thereby demonstrating that mitogen-induced enhancement of CFU-GEMMCSA production by leukemic B-lymphocytes is independent of DNA synthesis . Conditioned media of leukemic B-lymphocytes stimulated the formation of human marrow granulocyte-macrophage (GM-CFU) and early erythroid (BFU-E) colonies as well . These observations indicate a role for B-lymphocytes in the regulation of hemopoietic stem cell and progenitor cell growth in vitro. Acta Orthop Scand, 1987 Feb, 58(1), 4 - 13 Ultraclean air and antibiotics for prevention of postoperative infection . A multicenter study of 8,052 joint replacement operations; Lidwell OM et al.; To determine the value of ultraclean air in operating rooms, 8,052 operations for total hip- or knee-joint replacement were followed up for 1-4 years . For operations done in ultraclean air, bacterial contamination of the wound, deep joint sepsis, and major wound sepsis were substantially less than for operations done in conventionally ventilated rooms . Sepsis was also less frequent when prophylactic antibiotics had been given . The two precautions acted independently so that the incidence of sepsis after operation in ultraclean air and with antibiotics was much less than that when either was used alone . Wound sepsis was associated with an enhanced risk of joint sepsis . Staphylococcus aureus was the commonest joint pathogen, but infections with other organisms, often considered to be of low pathogenicity, were almost as numerous . Most S . aureus infections were traced to sources in the operating room. Immunopharmacology, 1987 Feb, 13(1), 73 - 86 Suppression of human lymphocyte responsiveness by forskolin: reversal by 12-O-tetradecanoyl phorbol 13-acetate, diacylglycerol and ionomycin; Shenker BJ et al.; Forskolin, a potent activator of adenylate cyclase, was examined for its ability to alter human peripheral blood lymphocyte (HPBL) activation by both mitogens and antigens . We found that forskolin, at concentrations ranging from 0.04 to 25 micrograms/ml, caused a dose-dependent inhibition of HPBL responses to mitogens (concanavalin A, phytohemagglutinin, pokeweed mitogen and Staphylococcus aureus) and to recall antigens (tetanus toxoid and streptokinase/streptodornase) . Inhibition was reflected in altered DNA, RNA and protein synthesis, including immunoglobulin production, and was not due to altered cell viability . Forskolin also induced a 19-fold increase in HPBL cyclic AMP levels at the same concentrations that suppressed HPBL function . To further define the mechanism(s) by which these elevations in cyclic AMP suppressed HPBL function, we tried to reverse these inhibitory effects with several agents; ascorbic acid, carbachol and levamisole had no effect . However, the phorbol ester, 12-O-tetradecanoyl phorbol 13-acetate, as well as L-alpha-1,2-dioleoyl diacylglycerol were able to completely reverse the inhibition . Furthermore, the Ca2+ ionophore, ionomycin, was also able to act synergistically with lower and less effective concentrations of 12-O-tetradecanoyl phorbol 13-acetate to reverse the inhibitory effects of forskolin . The data suggest that forskolin-induced elevations in cyclic AMP may lead to inhibition (or, more correctly, prevents the activation) of protein kinase C, presumably by inhibiting phospholipid turnover . Our studies suggest a linkage between these two opposing membrane-signal transduction systems with protein kinase C representing a pivotal point for various regulatory signals that ultimately control lymphocyte activation and function. Immunology, 1987 Feb, 60(2), 195 - 201 Frequency of B-lymphocyte transformation by Epstein-Barr virus decreases with entry into the cell cycle; Roome AJ et al.; The relationship between in vitro B-cell activation and transformation by Epstein-Barr virus (EBV) was studied . B cells were fractionated using discontinuous Percoll gradients to purify cells with resting morphology . Activation of resting cells for 24 hr with anti-Ig (mu chain specific) or Staphylococcus aureus Cowan I (SAC) resulted in transition of susceptible cells into the G1 phase of the cell cycle as shown by an increase in cell size, an increase in uridine incorporation and an increase in sensitivity to B-cell growth factor (BCGF) . Entry into S phase was achieved by extending the period of activation to 48-96 hr with high concentrations of SAC or anti-mu or using BCGF . SAC-activated cells entered S phase on Day 2 and anti-mu treated cells on Day 3 . Control (G0) cells and cell activated for varying lengths of time (G0/G1, G1/S) were exposed to EBV and plated in a limiting dilution assay to determine the frequency of EBV-transformable cells . Control cells and cells activated for 24 hr had a transformation frequency of 1-2% . With continued activation with SAC or anti-mu, however, transformation frequency decreased at a rate paralleling the entry of the population into S phase . Treating cells with low concentrations of anti-mu or SAC in combination with BCGF decreased the transformation frequency to levels lower than anti-mu or SAC alone, further suggesting that entry into S phase is accompanied by a reduction in transformability . These results indicate that resting B cells are highly susceptible to transformation, and that with in vitro activation into the cell cycle B cells become resistant to EBV transformation. J Dairy Res, 1987 Feb, 54(1), 1 - 5 Phage typing of Staphylococcus aureus associated with subclinical bovine mastitis; Mackie DP et al.; Six hundred and seventeen isolates of Staphylococcus aureus from subclinical clinical mastitis cases in 63 dairy herds in Northern Ireland were typed using a set of 25 phages . Ninety-four per cent of the isolates were typable, with nine phages, predominantly from groups I and III, being responsible for almost all of the lysis . Although 68 phage patterns were found, six of them typed 47.2% of the isolates . One strain accounted for 14.7% of the isolates, but the largest number of strains (44) was restricted to individual farms . The epidemiological significance of these findings for on-farm mastitis control is discussed. Clin Immunol Immunopathol, 1987 Feb, 42(2), 195 - 201 Immunoglobulin synthesis in myelodysplastic syndromes: normal B-cell and immunoregulatory T-cell functions; Merchav S et al.; Peripheral blood cells from patients with myelodysplastic syndromes were assayed for B-cell and immunoregulatory T-cell functions . The B/T cell ratio in myelodysplastic patients (n = 11) was significantly higher than in controls (n = 12) . These patients had a reduction in total T-cell (OKT3+) frequency and in T-cell subset (OKT4+/OKT8+) ratios . The response of patients' cells to both pokeweed mitogen (PWM) and phytohemagglutinin (PHA) was reduced, but patients' B cells responded normally to stimulation with Staphylococcus aureus Cowan (SAC) . The levels of IgG and IgM detected in 7-day culture supernatants of PWM-stimulated patient and control cells were similar . Normal B-cell and immunoregulatory T-cell functions were subsequently demonstrated in allogeneic co-culture combinations of enriched T and B cells from patients and controls . The data presented indicate that the frequent infections of myelodysplastic patients are not causally related to impaired humoral mechanisms . The data also favor the possibility that the stem cell disorder in these syndromes is functionally expressed at a subsequent stage to the lymphoid differentiation pathway. J Neurochem, 1987 Feb, 48(2), 455 - 62 Two monoclonal antibodies recognize Alzheimer's neurofibrillary tangles, neurofilament, and microtubule-associated proteins; Ksiezak-Reding H et al.; Two monoclonal antibodies that recognize Alzheimer's neurofibrillary tangles (ANTs), AD10 and AB18, have been characterized by immunoblotting against human and calf spinal cord neurofilament (NF) and calf brain microtubule preparations . Both antibodies bind to the 200-kilodalton (kd) (NF-H) and 160-kd (NF-M) but not to the 68-kd (NF-L) NF triplet proteins . They also bind to high-molecular-weight microtubule-associated proteins (MAPs) and tau . AD10 immunostains MAP2 and MAP1 families, whereas AB18 stains mainly MAP1 bands . Preincubation of intact filament preparation or nitrocellulose strips containing electroblotted NF proteins with Escherichia coli alkaline phosphatase completely blocks AD10 binding and partially blocks binding of AB18 . These results suggest that the determinants recognized by these antibodies are phosphorylated . Immunoblotting of peptide fragments generated by limited proteolysis of NF proteins with alpha-chymotrypsin and Staphylococcus aureus V8 protease shows that the localization of the antigenic determinants to AD10 and AB18 in NF-H is approximately 100 and 60 kd, respectively, away from the carboxy terminal, a region previously shown to form the NF projection side arm . In NF-M, the antigenic determinants to both antibodies are located also in the projection side arm, in a 60-kd polypeptide adjacent to the alpha-helical filament core . The results show that ANTs contain at least two phosphorylated antigenic sites that are present in NF and MAPs, a finding suggesting that ANTs may be composed of proteins or their fragments with epitopes shared by cytoskeletal proteins. Biochemistry, 1987 Jan 27, 26(2), 338 - 43 Functional role of proteolytic cleavage at arginine-275 of human tissue plasminogen activator as assessed by site-directed mutagenesis; Tate KM et al.; Activation of the zymogen form of a serine protease is associated with a conformational change that follows proteolysis at a specific site . Tissue-type plasminogen activator (t-PA) is homologous to mammalian serine proteases and contains an apparent activation cleavage site at arginine-275 . To clarify the functional consequences of cleavage at arginine-275 of t-PA, site-specific mutagenesis was performed to convert arginine-275 to a glutamic acid . The mutant enzyme (designated Arg-275----Glu t-PA) could be converted to the two-chain form by Staphylococcus aureus V8 protease but not by plasmin . The one-chain form was 8 times less active against the tripeptide substrate H-D-isoleucyl-L-prolyl-L-arginine-p-nitroanilide (S-2288), and the ability of the enzyme to activate plasminogen in the absence of fibrinogen was reduced 20-50 times compared to the two-chain form . In contrast, one-chain Arg-275----Glu t-PA has equal activity to the two-chain form when assayed in the presence of physiological levels of fibrinogen and plasminogen . Fibrin bound significantly more of the one-chain form of t-PA than the two-chain form for both the wild-type and mutated enzymes . One- and two-chain forms of the wild-type and mutated plasminogen activators slowly formed complexes with plasma protease inhibitors, although the one-chain forms showed decreased complex formation with alpha 2-macroglobulin . The one-chain form of t-PA therefore is fully functional under physiologic conditions and has an increased fibrin binding compared to the two-chain form. J Biol Chem, 1987 Jan 25, 262(3), 1312 - 9 Immunochemical detection of unique proteolytic fragments of the chick 1,25-dihydroxyvitamin D3 receptor . Distinct 20-kDa DNA-binding and 45-kDa hormone-binding species; Allegretto EA et al.; We have characterized proteolytic fragments of the chick intestinal 1,25-dihyroxyvitamin D3 (1,25-(OH)2D3) receptor, produced through either exogenous or endogenous protease action, utilizing a variety of physical and functional assays coupled to immunoblot detection methodology . Treatment of intestinal cytosol with increasing concentrations of trypsin resulted in a progressive diminishment of the 60-kDa receptor concomitant with the appearance of a 20-kDa fragment reactive by Western blot analysis to an anti-1,25-(OH)2D3 receptor monoclonal antibody . Cleveland analysis supported the receptor-origin of this 20-kDa fragment: a common immunoreactive species of 12 kDa could be generated by Staphylococcus aureus V8 protease treatment of the intact 60-kDa receptor as well as the 20-kDa proteolytic product . The 20-kDa fragment did not bind hormone but was capable of interacting with DNA-cellulose in a fashion identical to that of the 60-kDa receptor and, therefore, may contain the functional DNA-binding domain of the chick 1,25-(OH)2D3 receptor . Thus, this fragment likely represents the complement of a larger hormone-bound fragment that we have previously described (Allegretto, E . A., and Pike, J.W . (1985) J . Biol . Chem . 260, 10139-10145) . In contrast to the exogenous effect of trypsin, incubation of cytosol resulted in the time-dependent formation of an endogenous protease-derived fragment of 45 kDa . Cleveland analysis was consistent with the 60-kDa receptor derivation of the 45-kDa fragment . This species retained the hormone-binding site and the antibody determinant but was devoid of DNA-binding activity . Moreover, it generated neither the trypsin-dependent 20-kDa fragment nor the V8 protease-dependent 12-kDa species and, therefore, was derived from the opposite end of the receptor molecule . These data have facilitated the construction of a schematic model of the chick receptor in which the immunoreactive epitope is located between the functional domains for hormone binding and DNA binding. Biochim Biophys Acta, 1987 Jan 19, 927(1), 18 - 25 Influence of hypertonic solutions on catecholamine release from intact and permeabilized cultured chromaffin cells; Ladona MG et al.; Chromaffin cells purified from bovine adrenal medulla and maintained in primary culture were used to study the effects of hyperosmolarity on the nicotine- and high potassium-induced secretory response . A similar study was also performed on cells permeabilized with digitonin and with alpha-toxin from Staphylococcus aureus . Hyperosmolarity does not affect the spontaneous release of catecholamines from either intact cells or permeabilized cells . The nicotine-induced secretion and high potassium-induced secretion from intact cells are inhibited by hypertonic solutions; a 100% inhibition of net release was observed at 660 mOsm (sucrose as osmotic agent) . Veratridine- and the cation ionophore X537-A-induced release were both depressed under hyperosmotic conditions . Hyperosmolarity was shown to have reversible effects on the secretory response of intact cells . Finally, hyperosmolarity has intracellular effects on catecholamine release evoked by calcium from both detergent- and alpha-toxin-permeabilized cells . Our data show that hyperosmolarity has multiple effects on the cell membrane and the protein constituents associated with it, but has also a significant effect on intracellular reactions concerned with exocytosis. Eur J Biochem, 1987 Jan 15, 162(2), 279 - 86 Distribution of high-density lipoprotein 2 and 3 constituents during in vitro phospholipid hydrolysis; Perret BP et al.; Human high-density lipoproteins HDL2 (d = 1.068-1.125) and HDL3 (d = 1.125-1.210) doubly labelled with {3H}cholesterol/cholesteryl ester and with {acyl-14C}phosphatidylcholine were further incubated with phospholipases . Highly purified phospholipase A2 from Crotalus adamanteus allowed gradual degrees of lipolysis (30-90%) on both HDL2 and HDL3 . Moderate phospholipid hydrolyses were achieved using hepatic triacylglycerol lipase, partially purified from post-heparin plasma . Moreover, the latter enzyme seemed to exert a lysophospholipase activity, acting on the 2-acyl-sn-glycero-3-phosphocholine generated . A purified sphingomyelinase C from Staphylococcus aureus was also used and completely hydrolysed HDL sphingomyelin . After incubation, doubly labelled HDL2/HDL3 were reisolated in their appropriate density interval . In the presence of albumin, which bound most of the lipolysis products, phospholipolysis induced a phospholipid depletion of the particles and a heterogeneous partition of all HDL2 constituents between the HDL2 and HDL3 density intervals . Radioactivity distributions correlated with mass movements . The 'HDL3-like' particles isolated after HDL2 lipolysis were twice as rich in cholesterol as plasma HDL3 . No loss of apoprotein A1 was recorded due to phospholipolysis . In the absence of albumin, the density distributions of HDL2 or HDL3 constituents were unaffected by phospholipolysis, the products of lipolysis being reisolated with the stable particles . Control and treated HDL were also reisolated by equilibrium density gradient ultracentrifugation, gel chromatography or by gradient gel electrophoresis . Phospholipase treatment in the presence of albumin induced a shift of the HDL2 or HDL3 whole distribution towards particles of higher density and lower apparent size . Lipolysed HDL2 thus showed characteristics intermediate between those of HDL2 and HDL3 . So, phospholipolysis may affect the physical parameters of HDL particles, but additional pathways such as cholesterol movements and apoprotein loss must be linked to achieve the HDL2----HDL3 interconversion. S Afr Med J, 1987 Jan 10, 71(1), 25 - 7 Right-sided valvular infective endocarditis . A clinicopathological study of 29 patients; van der Westhuizen NG et al.; Out of 29 patients with right-sided infective endocarditis 13 had associated congenital heart disease and none was a drug addict . Staphylococcus aureus was the commonest organism isolated . The tricuspid valve was infected twice as commonly as the pulmonary valve . Only 11 out of the 29 patients had valvular infection potentially amenable to surgical therapy at the time of death . Ring abscesses were present in 10 patients. J Biol Chem, 1987 Jan 5, 262(1), 176 - 81 Purification of a beta 35 form of the beta gamma complex common to G-proteins from human placental membranes; Evans T et al.; The significance of the 36,000-35,000-dalton doublet of proteins referred to as the beta-subunit of the G-proteins remains unresolved . An immunological distinction between the 36,000 (beta 36)- and 35,000 (beta 35)-dalton proteins has been reported (Roof, D . J., Applebury, M . L., and Sternweis, P . C . (1985) J . Biol . Chem . 260, 16242-16249; Mumby, S . M., Kahn, R . A., Manning, D . R., and Gilman, A . G . (1986) Proc . Natl . Acad . Sci . U.S.A . 83, 265-269) . The availability of homogeneous preparations of beta 36 and beta 35 will facilitate studies designed to address the significance of the doublet structure . This manuscript presents the preparative purification of the beta 35 protein from a highly enriched source, human placenta . Unlike the beta gamma complex associated with G-proteins from placenta, the beta 35 preparations consist predominantly of the 35,000-dalton protein . The gamma-peptide associated with beta 35 is indistinguishable electrophoretically and immunologically from that associated with the placental G-protein oligomers . The beta 35 preparation and beta-subunit doublet exhibit similar specific activities in inhibiting human platelet adenylate cyclase activity . The preparations have proven useful for the generation of a panel of rabbit polyclonal antisera that recognize beta 35, Gt beta 36, and both beta 36 and beta 35 in the doublet associated with Gs, Gi, Go, and Gp . One antiserum generated to beta 35 recognizes the gamma-peptide associated with these preparations and human placental G-protein oligomers . The antiserum does not recognize Gt gamma . Protease digestion of Gt beta 36 and human placental beta 35 with Staphylococcus aureus V8 protease identified a unique peptide generated from beta 35 which is absent in beta 36 digests. Appl Environ Microbiol, 1987 Jan, 53(1), 88 - 91 Lysostaphin lysis procedure for detection of Staphylococcus aureus by the firefly bioluminescent ATP method; Tuncan EU et al.; The objective of this study was to examine the use of lysostaphin as an ATP-extracting agent for the estimation of Staphylococcus aureus cell number by a rapid bioluminescent ATP method . The results of the study showed that lysostaphin (22 U/ml) was able to lyse most of the S . aureus cells (greater than 99.9%) at room temperature in 1 min; ATP of S . aureus cells extracted by the lysostaphin lysis procedure was stable for 24 h in the presence of EDTA; there was a linear relationship between the ATP content and the number of S . aureus cells (ranging from 10(4) to 10(6) CFU/ml); and the lysis of S . aureus cells by lysostaphin allowed estimation of the number of S . aureus cells in mixed cultures and in meat samples. J Am Acad Dermatol, 1987 Jan, 16(1 Pt 2), 227 - 9 Toxic shock syndrome as a complication of dermatologic surgery; Huntley AC et al.; We report a case of toxic shock syndrome occurring after an excisional skin biopsy in an otherwise healthy woman . Her clean-appearing wound grew Staphylococcus aureus and was implicated as the source of toxin . Dermatologists should be aware that toxic shock syndrome may occur as a complication of simple dermatologic surgery. J Am Acad Dermatol, 1987 Jan, 16(1 Pt 2), 205 - 8 Catheter infections during continuous peritoneal dialysis; Dale SE et al.; A typical tunnel infection is described in a patient receiving continuous ambulatory peritoneal dialysis . This complication of peritoneal dialysis is important to study and manage well because the infection leads to removal of the catheter and interruption of therapy . Most infections are due either to Staphylococcus aureus or to S . epidermidis. J Clin Microbiol, 1987 Jan, 25(1), 87 - 90 Investigation by syringe method of effect of tampons on production in vitro of toxic shock syndrome toxin 1 by Staphylococcus aureus; Lee AC et al.; A syringe method was designed to test the effect of tampons on the growth of three toxic shock syndrome-associated strains of Staphylococcus aureus and their in vitro production of toxic shock syndrome toxin 1 (TSST-1) under different conditions . Various amounts of TSST-1 were recovered from different tampons inoculated with these strains . Generally, the addition of 10% porcine blood to the growth medium, incubation in the presence of 5% CO2, or the combination of these two factors resulted in the stimulation of TSST-1 production. Crit Care Med, 1987 Jan, 15(1), 47 - 50 Broviac catheterization in low birth weight infants: incidence and treatment of associated complications; Sadiq HF et al.; Fifty-two Broviac catheters were inserted in 40 preterm and eight term infants for 1733 days of catheter use . Thirty-six (69%) catheters were associated with complications of infection and/or thrombosis, a complication rate of 1/48 catheter days . The patients who developed complications were of a significantly lower gestational age and had a lower mean birth weight when compared with those who developed no complications . The incidence of catheter-related sepsis was 69% in the very low birth weight infants and only 20% in the infants with birth weights over 1500 g . Eighteen of the 26 catheter-associated infections were treated with antibiotics without catheter removal . Successful resolution of the infections with retention of the catheter occurred in 14 of the 18 episodes . Infections with Staphylococcus aureus constituted three of four treatment failures . Urokinase infusion was successful in causing thrombolysis in eight of the nine cases . Broviac catheters in neonates, and especially in preterm infants under 1500 g, are associated with a high incidence of complications . Our experience indicates that some complications can be selectively managed without sacrificing the venous access. Infection, 1987, 15(5), 351 - 3 Toxic shock syndrome toxin-1 (TSST-1): influence of its production by subinhibitory antibiotic concentrations; Dickgiesser N et al.; Using seven different toxic shock syndrome toxin-1 (TSST-1) producing Staphylococcus aureus strains, we examined the influence on growth and toxin production of subinhibitory concentrations of clindamycin, erythromycin, lincomycin, kanamycin, tetracycline and tunicamycin . The behaviour of six S . aureus (= W/MT-strains) was identical, the one of S . aureus MN8 was different in part . Using the different subinhibitory antibiotic concentrations, bacterial growth was inhibited by tunicamycin only . Toxin production was influenced by clindamycin, erythromycin, lincomycin, kanamycin and tetracycline without simultaneous changes in the number of cells; MN8 was more sensitive to clindamycin and lincomycin than W/MT strains . Very small differences or no differences at all were found between the two bacterial groups in experiments using erythromycin, kanamycin and tetracycline . Tunicamycin caused elevated TSST-1 concentrations of 100% (MN8) or 65% (W/MT) above the control level if used at concentrations of 4 mg/l or 16 mg/l respectively; this result is interpreted as a higher output of TSST-1 caused by the damage to the bacterial cell wall . From these results it is not possible to conclude that different mechanisms of regulation of TSST-1 expression exist between S . aureus MN8 and the other TSST-1 positive S . aureus strains. Gene, 1987, 58(1), 87 - 97 A gene fusion system for generating antibodies against short peptides; Lowenadler B et al.; A novel method to obtain specific antibodies against short peptides is described, involving synthesis of the corresponding oligodeoxynucleotides followed by cloning into a new set of fusion vectors, pEZZ8 and pEZZ18, based on two synthetic IgG-binding domains (ZZ) of Staphylococcus aureus protein A . The soluble gene fusion product thus obtained, can be collected from the culture medium of Escherichia coli and rapidly recovered in a one-step procedure by IgG affinity chromatography . The system was used to express a fusion protein consisting of the two Z fragments and the C-terminal part {amino acids (aa) 57-70} of human insulin-like growth factor I (IGF-I) . This 16-kDa protein was purified by affinity chromatography on IgG Sepharose and antibodies were raised in rabbits . The fusion protein elicited peptide-specific antibodies, as measured by solid-phase radioimmuno assay and Western blotting, reactive with both synthetic C-terminal peptide and the native human IGF-I protein . The results suggests that the gene fusion system can be used for efficient antibody production against short peptides encoded by synthetic oligodeoxynucleotides. Acta Med Scand, 1987, 222(2), 175 - 82 Antibiotic treatment of Staphylococcus aureus endocarditis . A review of 119 cases; Frimodt-Moller N et al.; Clinical and bacteriological information of Staphylococcus aureus endocarditis was reviewed in 119 cases from all over Denmark . Overall mortality was 71% . Survival correlated with antistaphylococcal treatment, short duration from onset of infection to start of treatment, and long duration of treatment . In spite of relevant treatment, mortality was significantly lower in cases infected with penicillin-susceptible strains than when penicillin-resistant strains were isolated . There were no differences in the effect of various anti-staphylococcal treatment regimens; in particular, there were no differences in mortality with regard to beta-lactam antibiotics alone as compared to beta-lactam antibiotics in combination with aminoglycosides . However, embolic manifestations occurred more often after start of treatment with combination therapy than with beta-lactam antibiotics alone. Vet Med Nauki, 1987, 24(6), 33 - 8 {Drug resistance and virulence of Staphylococcus aureus strains isolated from cows with endometritis}; Korudzhiiski N et al.; A study was carried out on the drug resistance as associated with the virulence of Staphylococcus aureus strains isolated from cows with endometritis . Hirota's method was employed to ascertain that resistance to penicillin, erythromycin, neomycin, and kanamycin was governed by plasmids . It was found at the same time that, fairly often, with the elimination of some of the resistant markers the virulence of the strain variants also dropped . However, these two properties did not correlate with each other. Chemotherapy, 1987, 33(5), 361 - 75 Gentamicin- and methicillin-resistant Staphylococcus aureus: in vitro susceptibility to antimicrobial drugs; Traub WH et al.; Thirteen nosocomially significant, gentamicin- and methicillin-resistant (GRMR) Staphylococcus aureus isolates, all of phage group III/M (lysotype 42E/47/53/54/75/77/83A/84/85/94/96), were uniformly resistant against augmentin, erythromycin, fosfomycin, gentamicin, methicillin, oxacillin, penicillin G, tetracycline, and tobramycin, but differed in susceptibility to cefamandole, ciprofloxacin, clindamycin, imipenem, josamycin, the synthetic chinolone Ro 23-6240, and ofloxacin . All isolates were susceptible to chloramphenicol, coumermycin, fusidic acid, novobiocin, rifampin, teicoplanin, trimethoprim-sulfamethoxazole (cotrimoxazole), and vancomycin . One isolate was of intermediate susceptibility to netilmicin . On a weight-for-weight basis, the 7 most active drugs were rifampin, coumermycin, cotrimoxazole, novobiocin, teicoplanin, fusidic acid, and vancomycin (in decreasing order) in terms of minimal inhibitory concentrations . With regard to minimal bactericidal concentrations, coumermycin, rifampin, vancomycin, teicoplanin, cotrimoxazole, ofloxacin, and ciprofloxacin (in decreasing order) were the 7 most potent antimicrobial drugs . Freshly defibrinated human blood {65% (v/v)} combined with chloramphenicol and rifampin, respectively, resulted in a weak additive effect (time kill curves) . Indifferent effects were observed following combination of blood with ciprofloxacin, cotrimoxazole, coumermycin, fusidic acid, imipenem, netilmicin, novobiocin, ofloxacin, compound Ro 23-6240, teicoplanin, and vancomycin . Rifampin combined with novobiocin, teicoplanin, and vancomycin, respectively, in the presence of 65% (v/v) human blood, resulted in an additive effect . Combinations of rifampin with 9 other antimicrobial drugs in blood yielded essentially indifferent effects. Chemotherapy, 1987, 33(5), 331 - 9 Effect of antibiotic protein binding on the killing rate of Staphylococcus aureus and on the paradoxical phenomenon; Odenholt I et al.; The killing kinetics of Staphylococcus aureus, exposed to various concentrations of dicloxacillin in broth and in broth with 40 g/l human albumin was studied . When the free concentrations of dicloxacillin were identical in the two media, no difference in killing capacity could be demonstrated at concentrations above MIC, indicating that only the free antibiotic fraction is antibacterially active . However, at concentrations identical to the MIC, a better bactericidal effect in the medium containing albumin was found . In experiments where equal total concentrations were compared in the two media, an increasing bactericidal effect in the medium containing albumin could be demonstrated at concentrations between 10-100 X MIC . The most probable explanation for this was a prominent paradoxical effect with increasing antibiotic concentrations on the killing rate of S . aureus in broth . This effect was neutralized in the presence of albumin due to the lower free antibiotic concentration in this medium. Infect Control, 1987 Jan, 8(1), 30 - 3 Staphylococcus aureus colonization and infection after discharge from a term newborn nursery; Wang EE et al.; During a 6-month period, term infants underwent nasal and umbilical swabbing within 96 hours of delivery and again at 1 to 18 (mean 3.8) weeks after discharge . Swabs were inoculated onto horse blood agar and all S aureus isolates were phage typed . Two hundred three infants were enrolled and follow-up was obtained for 181 (89%) . Thirty-two of 181 (17.6%) were initially colonized, of whom 12 (37.5%) were colonized on follow-up . Thirty-two of 181 became colonized subsequent to hospital discharge . No single phage type predominated . Twenty-five patients were diagnosed to have clinical illness--omphalitis (11), conjunctivitis (10), and pustulosis (5) . Six of these were colonized with S aureus initially, although organisms of the same phage type were not recovered from cultures obtained at the time of clinical illness . There was no significant difference in the rate of infections in colonized (19%) versus non-colonized (12%) infants . In five patients where S aureus was recovered at the time of symptoms, all organisms were acquired subsequent to discharge . We conclude that nursery colonization with S aureus did not lead to clinical illness, and clinical illnesses previously ascribed to S aureus frequently occur in the absence of these bacteria. Infect Control, 1987 Jan, 8(1), 24 - 9 Methicillin-resistant Staphylococcus aureus in a nursing home; Storch GA et al.; During a 13-month period, 25 residents of a nursing home were found to have positive cultures for methicillin-resistant Staphylococcus aureus (MRSA), including 17 with clinically significant infections . The outbreak came to attention in February 1985 when pneumonia was diagnosed in five residents during a 10-day period, and sputum cultures from all five were positive for MRSA . A survey revealed that nine (12%) of a sample of 74 residents and nine (7%) of 130 personnel had positive cultures for MRSA . Six of nine residents with MRSA detected in the culture survey had not been hospitalized for 6 or more months before the survey, suggesting acquisition of MRSA in the nursing home . Implementation of control measures was associated with a decreased occurrence but not complete elimination of new cases . MRSA in nursing homes is of concern because these institutions might serve as reservoirs for MRSA in the community . Further studies are required to define the magnitude of the problem, as well as optimal control measures. Infect Control, 1987 Jan, 8(1), 15 - 23 Outbreak due to methicillin- and rifampin-resistant Staphylococcus aureus: epidemiology and eradication of the resistant strain from the hospital; Bitar CM et al.; A methicillin- and rifampin-resistant strain of Staphylococcus aureus was introduced into a university hospital by interstate transfer of an infected surgical patient . An outbreak occurred, and 17 patients became infected or colonized with the epidemic strain . Reservoirs appeared to be patients who were infected or colonized with the resistant S aureus and possibly two nurses who were nasal carriers . The outbreak isolate was likely spread by contact with contaminated hands of personnel . A retrospective case-control study identified tracheostomy, debridement, and irrigation of wounds by power spray and prolonged nasogastric intubation as risk factors for acquisition of the epidemic strain . Analysis of factors by groups indicated that surgical procedures, wound care procedures and instrumentation of the respiratory tract were significantly associated with cases . The nasal carrier state was eradicated in two nurses by topical application of 5% vancomycin . The epidemic strain was eradicated from the hospital 8 months after it was introduced. Arch Dis Child, 1987 Jan, 62(1), 82 - 4 Methicillin resistant Staphylococcus aureus in milk; Parks YA et al.; Three separate outbreaks of gentamicin and methicillin resistant Staphylococcus aureus in a special care baby unit are described . The outbreaks ceased only after a milk bank worker was identified as a carrier of the strain . It is postulated that the infant milk feeds served as a vehicle of spread. Acta Neurochir (Wien), 1987, 86(3-4), 83 - 8 Intrathecal administration of netilmicin in gentamicin-resistant ventriculitis; Donauer E et al.; In ventriculitis caused by Gentamicin-resistant staphylococcus aureus and staphylococcus epidermidis, Netilmicin was administered intrathecally to 19 patients under continuous control of the Netilmicin concentration in cerebro-spinal fluid (CSF) . This therapy was able to bring these otherwise lethal infections under control, usually within 10 days . Pharmacokinetic studies with different doses have shown that doses of 2 X 3 mg are to be recommended in moderately severe cases of ventriculitis, and in most severe infections 3 X 3 mg daily intraventricularly for adults . In infants daily intraventricular injections of 2 X 0.4-0.5 mg Netilmicin are a sufficient dose to produce an effective antibiotic concentration level. Diagn Clin Immunol, 1987, 5(2), 62 - 8 A simultaneous flow cytometric measurement of neutrophil phagocytosis and oxidative burst in whole blood; Trinkle LS et al.; To quantitate phagocytosis and respiratory burst by polymorphonuclear leukocytes (PMNs), 1-ml aliquots of human whole blood were incubated with 50 microM dichlorofluorescin diacetate for 10 min at 37 degrees C prior to the addition of Texas Red-labelled Staphylococcus aureus (SA) . At 5-min intervals PMNs were analyzed by flow cytometry for ingested SA and resultant production of hydrogen peroxide, an oxidative burst metabolite . PMNs from infected patients or normals that had been primed for 1 hr with bacterial endotoxin displayed enhanced phagocytosis and oxidative burst activity compared to controls . PMNs from a patient with chronic granulomatous disease were less reactive than those of normals . The whole blood analysis is a simple, rapid, and inexpensive technique for the clinical evaluation of PMN function in a physiologic milieu. Scand J Infect Dis, 1987, 19(2), 195 - 203 Staphylococcus aureus bacteremia: significance of hyperbilirubinemia; Watanakunakorn C et al.; We studied 233 consecutive episodes of Staphylococcus aureus bacteremia in 230 patients between 1980 and 1984 at a community teaching hospital . Bacteremia was community-acquired in 78 episodes, acquired from nursing homes in 22 episodes and hospital-acquired in 133 episodes . The over-all mortality was 48.9% . Patients greater than or equal to 60 years had higher mortality (62.0%) than patients less than 60 years old (25.3%) . Hospital-acquired bacteremia was associated with a higher mortality (59.4%) than community-acquired bacteremia (29.5%) . The respiratory tract as the portal of entry of bacteremia was associated with a higher mortality (80.4%), as compared to 53.5% when the portal of entry was undetermined, and 28.1% when the portal of entry was other sources . Increasing serum creatinine levels were associated with increasing mortality: less than 88.4 mumol/l (26.5%), 97.2-168.0 mumol/l (51.1%), and greater than 176.8 mumol/l (67.9%) . Increasing serum bilirubin levels were also associated with increasing mortality: less than 17.1 mumol/l (40.6%), 18.8-49.6 mumol/l (57.1%), and greater than 51.3 mumol/l (84.2%) . Very high leukocyte counts were associated with higher mortality: greater than 20 X 10(9)/l (73.9%), 10-20 X 10(9)/l (37.4%), and less than 10 X 10(9)/l (45.8%). Chemotherapy, 1987, 33(4), 243 - 9 Clindamycin at subinhibitory concentrations enhances antibody- and complement-dependent phagocytosis by human polymorphonuclear leukocytes of Staphylococcus aureus; Veringa EM et al.; The influence of subinhibitory concentrations of clindamycin on opsonization and phagocytosis of Staphylococcus aureus was studied . S . aureus was grown overnight in the presence or absence of one half or one quarter of the minimal inhibitory concentration (MIC) of clindamycin . Radioactively labeled S . aureus was opsonized for various periods of time in different concentrations of normal serum, heated antiserum and serum of patients with agammaglobulinaemia or C3 deficiency . Complement- as well as antibody-dependent phagocytosis of the antibiotic treated S . aureus was significantly enhanced, compared to phagocytosis of the untreated control . Killing experiments showed that clindamycin-treated S . aureus was also better killed by the granulocytes than untreated S . aureus . The mechanism of action is likely to be an increased susceptibility of clindamycin-treated bacteria to antibody- and complement-dependent phagocytosis. Dev Comp Immunol, 1987 Winter, 11(1), 191 - 202 Partial characterization of chicken spleen cell culture supernatants stimulated with Staphylococcus aureus; Schoof DD et al.; The role of accessory cells in the proliferative response of chicken spleen cells to Staphylococcus aureus Cowan I (SAC) was examined . It was found that chicken spleen cells cultured with SAC produced a soluble molecule capable of causing proliferation when culture supernatants were added to spleen cells . The molecules responsible for this activity were stable in terms of exposure to extremes of heat and pH . Gel filtration of culture supernatants revealed biological activity, over a broad range of molecular weights, as measured by spleen cell proliferation . Similar findings were obtained when SAC was sonicated and evaluated following gel filtration . Exposure of culture supernatants to trypsin abrogated biological activity . The pivotal role of adherent cells in the generation of biologically active molecules is suggested by the ability of peritoneal exudate cells incubated with SAC to produce biologically active supernatants . In addition, the proliferative response of spleen cells to SAC was sensitive to chloroquine. Vutr Boles, 1987, 26(1), 33 - 7 {Analysis of the autopsy data on 48 cases of infectious endocarditis in patients over 14}; Prodanov A et al.; Forty eight cases with infections endocarditis (IE) are analyzed--27 males and 21 females (75% at the age of 31-60) . Only in 12.5% IE has developed on intact valvular apparatus . In 83.4% the preceding valvular pathology is rheumatic heart defect (RHD), and congenital cardiac malformations and valvular dysplasia--8.3% each . The mitral-aortic valvular defects predominate in RHD, followed by aortic, tricuspid and mitral ones . In 83.3% X IE is with valvular localization, in 4.2%--parietal and in 12.5%--on valvular prostheses . Vegetations are highly specific for valvular IE (100%) as well as ulcerations (90%) and perforations (45%) . Ruptures of extra-valvular structures are established in 8.3% . The inflammatory alterations in myocardium are in 62.5% . The pericardial changes are found in 14.6%, mainly in the cases with valvular prosthesis . Septic changes are established in all cases in the spleen and regeneration of bone marrow, and vascular changes--in 85.4% . The cases with embolism and infarctions predominate in the kidneys (79.1%), followed by the cases with Lohlein's focal nephritis (18.6%) . The inflammatory changes in liver are rare--6.9% . In 37.5% changes in brain were established, embolism according to incidence being first, followed by cerebral hemorrhages, cerebral abscesses and purulent leptomeningitis Staphylococcus aureus proved to be most frequent causative infectious agent established et necropsy. Microbiol Immunol, 1987, 31(1), 35 - 44 Electron microscopic demonstration and isolation of ribosomes in mesosomes from Staphylococcus aureus; Nakasone N et al.; Mesosomes of Staphylococcus aureus 209P were observed to be extruded as tubules upon protoplast formation by electron microscopy and isolated under hypertonic conditions to maintain their structural integrity by differential centrifugation followed by sucrose density gradient centrifugation . Isolated mesosomes were composed of long, branched tubules of irregular sizes and they were shortened during purification . Thin sections of isolated mesosomes showed that the mesosomal tubule was surrounded by a triple-layered membrane and contained ribosome-like particles in diameter of about 15 to 20 nm . These particles were isolated from purified mesosomal preparation by disrupting the mesosomal tubule with deoxycholate and Triton X-100 under hypotonic conditions followed by a linear sucrose density gradient centrifugation . Negatively stained preparations of the isolated particles revealed the same appearance as those of the ribosomes isolated from the cytoplasm . The mesosomal particles sedimented at 70S in sucrose gradients in the presence of 10 mM Mg2+, but they were dissociated into two subparticles, 50S and 30S subunits, upon lowering the Mg2+ concentration to 1 mM . These findings indicate that the mesosomal tubule is packed with ribosomes. Avian Dis, 1987 Jan-Mar, 31(1), 140 - 4 Preliminary studies on bacterial interference of staphylococcosis of chickens; Nicoll TR et al.; Previous studies indicated that Staphylococcus epidermidis strain 115 might be an effective interfering agent in reducing the rate of staphylococcosis in turkeys . In the present study, strain 115 was avirulent when administered to 3-day-old chicks by oral, aerosol, or intravenous route . Strain 115 adhered specifically to tracheal, lung, air-sac, and liver cells in vitro and interfered with subsequent colonization by virulent Staphylococcus aureus . In vivo colonization of lungs and livers of young chicks occurred following exposure to aerosols of strain 115 . Strain 115 interfered with the in vivo colonization of lungs and livers by virulent S . aureus. J Biochem (Tokyo), 1987 Jan, 101(1), 217 - 23 Positions of disulfide bonds in riboflavin-binding protein of hen egg white; Hamazume Y et al.; Riboflavin-binding protein of hen egg white (egg-white RBP) comprised 219 amino acid residues and nine disulfide bonds . To identify the locations of these bonds, the native protein was oxidized with cyanogen bromide and digested with trypsin, thermolysin, and Staphylococcus aureus V8 protease . The cystine-containing peptides were isolated by HPLC . Amino acid analyses and amino acid sequence analyses of the reduced pyridylethylated derivatives of the cystine peptides showed that seven of the disulfide bonds were as follows: Cys(24)-Cys(73), Cys(57)-Cys(138), Cys(64)-Cys(110), Cys(99)-Cys(169), Cys(116)-Cys(134), Cys(103)-Cys(152), Cys(167)-Cys(202) . The other two disulfide bonds were either Cys(5)-Cys(32) and Cys(33)-Cys(77) or Cys(5)-Cys(33) and Cys(32)-Cys(77). Antimicrob Agents Chemother, 1987 Jan, 31(1), 16 - 20 Oxacillin killing curve patterns of Staphylococcus aureus isolates by agar dilution plate count method; Woolfrey BF et al.; The bactericidal dynamics of oxacillin against four Staphylococcus aureus isolates with known 24-h "persister" percentages were studied by using the agar dilution plate count method . Isolates were selected to provide a representative spectrum whose individual 24-h trough intrinsic persister percentages ranged from greater than 1 to less than 0.01% . Resultant agar dilution plate count method killing curve patterns were found to be reproducible and served to characterize each isolate . The paradoxical effect was observed for each isolate, with paradoxical peaks tending to develop and diminish sequentially during the course of oxacillin action . The observed strain-dependent dynamics of oxacillin killing underscore the artifactual nature of the so-called tolerance phenomenon and negate the usefulness of MBCs and MBC/MIC ratios for the characterization of S . aureus isolates. Scand J Infect Dis, 1987, 19(1), 137 - 40 Successful non-surgical management of pyogenic liver abscess; Loh R et al.; A 5-month-old infant presented with high fever, irritability, and poor feeding . Examination revealed an enlarged liver and neutropaenia . Ultrasonography was unhelpful, but a diagnosis of liver abscess was confirmed by computed tomography on day 4 of the illness . Gallium scan was normal on day 4 but showed a resolving liver abscess on day 6 . Staphylococcus aureus was grown in blood culture . The infant recovered with antibiotic treatment alone. Respiration, 1987, 51(1), 1 - 9 Influence of experimentally induced airway inflammation on the reactivity of cat lung parenchymal strips to histamine and acetylcholine; Banovcin P et al.; The reactivity of cat lung parenchymal strips to histamine and acetylcholine in vitro was studied at various stages of inflammation of the airways induced experimentally by the intratracheal administration of turpentine oil or Staphylococcus aureus, both in aerosol form . With histamine, the contractile responses of the lung strips were enhanced depending on the stage of turpentine oil inflammation . Similarly, the lung strips responded more to histamine in the staphylococcal inflammation . The reactivity of the lung strips to acetylcholine remained mainly unaffected by either type of inflammation. J Assoc Off Anal Chem, 1987 Jan-Feb, 70(1), 35 - 8 Most probable number method for isolation and enumeration of Staphylococcus aureus in foods: collaborative study; Lancette GA et al.; Enumeration of Staphylococcus aureus in foods was collaboratively studied by comparing the present AOAC final action method, 46.062, which uses trypticase soy broth with 10% NaCl to a proposed replacement method which uses the same broth with 1% sodium pyruvate added . Fifteen collaborators analyzed uninoculated samples of milk, tuna salad, and ground turkey, as well as samples inoculated with low (10(2) cells/g), middle (10(4) cells/g), and high (10(6) cells/g) levels of S . aureus . The samples were frozen immediately to maintain the inoculated level of S . aureus in the food . A different strain of S . aureus was used for each food; heat-stressed S . aureus cells were used to inoculate the milk samples . The pyruvate-amended broth significantly (alpha = 0.05) increased enumeration of low, middle, and high levels of S . aureus from milk and ground turkey, and from tuna salad at middle and high levels . The pyruvate-amended media method has been adopted official first action to replace method 46.062. Nippon Sanka Fujinka Gakkai Zasshi, 1987 Jan, 39(1), 99 - 105 {Antimicrobial effect of human endometrial peroxidase on Escherichia coli and Staphylococcus aureus}; Iwasaku K et al.; Human endometrium had potent peroxidase activity and an antimicrobial effect . Relationships which existed between the human endometrial peroxidase and antimicrobial effect were studied . Human endometrial peroxidase activity was measured by the modified method of Himmelhoch . In the normal menstrual cycle, the peroxidase activity in the secretory phase was higher than in the proliferative phase . The peroxidase activity in endometrial carcinoma was remarkably higher than in the normal menstrual cycle . The organisms (E.coli, S.aureus) and peroxidase extract were incubated for 60 min at 37 degrees C . Bacterial viability was determined by the plate culture method . The viable cell count was decreased . Endometrial peroxidase had an antimicrobial effect on E.coli and S.aureus . But the antimicrobial effect on clinically isolated S.aureus exerted by the peroxidase was not effective . Although the peroxidase activity in the endometrial carcinoma was higher than in the normal endometrium, there was no detectable antimicrobial effect . Peroxidase of endometrial carcinoma was suspected of having some different characteristics from normal endometrium. Am J Clin Pathol, 1987 Jan, 87(1), 94 - 100 Measurement of plasma fibrin D-dimer levels with the use of a monoclonal antibody coupled to latex beads; Greenberg CS et al.; Recently, monoclonal antibody (DD-3B6) to fibrin D-dimer was prepared and coupled to latex beads to provide a specific test (Dimertest) for fibrinolysis . The purpose of this study was to evaluate the Dimertest assay as a clinical laboratory test for the measurement of plasma fibrin D-dimer derivatives . The Dimer-test assay specifically detected 2 micrograms/mL of purified fibrin D-dimer or fibrin D-dimer/fragment E complex added to afibrinogenemic plasma but did not detect 500 micrograms/mL of either fibrinogen fragments X, D, E, or 160 micrograms/mL cross-linked fibrinogen . The fibrin(ogen) degradation product (FDP) assays of American Dade or Wellcome Diagnostics detected 5.0 micrograms/mL of fibrin D-dimer and from 1 to 10 micrograms/mL of the other FDPs . Twenty-eight percent of 150 random plasma samples assayed from hospitalized patients were positive for fibrin D-dimer derivatives . Plasma samples from 152 patients suspected of having disseminated intravascular coagulation (DIC) were assayed for serum FDP (Wellcome Diagnostics) and plasma fibrin D-dimer derivatives . Samples from 69% of patients with serum FDP levels less than 10 micrograms/mL, and more than 90% of those with serum FDP levels greater than 10 micrograms/mL, were positive for fibrin D-dimer derivatives . Dimertest results were not modified by heparin, streptokinase, freeze-thawing, or clotting plasma . Serum fibrinogen-related antigens were immunoadsorbed from Dimer-test positive sera by anti-fibrinogen antibody and formalin-fixed Cowan I strain Staphylococcus aureus . Analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and protein blotting with the use of monoclonal antibody DD-3B6 demonstrated a protein band with similar mobility to purified D-dimer . The measurement of plasma fibrin D-dimer derivatives by the Dimertest assay is a rapid, sensitive, and specific laboratory test for fibrinolysis . The Dimertest assay has proven to be a useful addition to the clinical laboratory and should be helpful in the diagnosis and management of patients with diseases associated with fibrinolysis. Blood, 1987 Jan, 69(1), 308 - 15 Proliferative and differentiative responses of B cells from human marrow graft recipients to T cell-derived factors; Matsue K et al.; Upon activation, B cells express growth and differentiation receptors that permit them to proliferate and differentiate . The aim of this study is to define the nature of the intrinsic B cell defects found in marrow recipients using assays for B cell activation, proliferation, and differentiation . B cells from five short-term (less than three months postgrafting) and 15 long-term (greater than one year postgrafting) marrow recipients (ten with and five without chronic graft-v -host disease {GVHD}) were studied . T cell supernatants (T-sup) were prepared by stimulating normal T cells with 12-0-tetradecanoyl-phorbol-13-acetate (TPA) and phytohemagglutinin . Highly purified B cells were used to assess B cell proliferation responses to T-sup after Staphylococcus aureus Cowan I (SAC) activation and for B cell immunoglobulin production responses to T-sup stimulation after SAC activation . B cells from all five short-term patients and one long-term patient with chronic GVHD did not respond to any stimulation . B cells from two patients with chronic GVHD responded to SAC but had decreased proliferative and differentiative responses to T-sup . B cells from three of seven patients with chronic GVHD and two of five long-term healthy patients could proliferate but could not secrete immunoglobulin in response to SAC plus T-sup stimulation . Furthermore, there was a significant correlation between serum IgG and/or IgM in marrow recipients and the differentiative responses of their B cells to T-sup (P = 0.0075, Fisher's Exact) . B cell defects occur at various stages of maturation postgrafting . These defects include the failure to respond to the SAC activation signal, the failure to proliferate in response to T-sup, and the failure to differentiate in response to T-sup . These findings are probably due to the inability of B cells from certain marrow recipients to undergo a second round of ontogeny. Blood, 1987 Jan, 69(1), 27 - 32 Topology and order of formation of interchain disulfide bonds in von Willebrand factor; Wagner DD et al.; Interchain disulfide bonds between the subunits in von Willebrand factor (vWf) dimers and in vWf multimers have been studied using some unique features of the cultured human umbilical vein endothelial cell system . Ammonium chloride inhibition of multimerization of vWf allowed selective examination of vWf dimeric molecules, and monoclonal antibody against the vWf propolypeptide was used to separate pro-vWf dimers from mature dimers . After cleavage of dimers and multimers with Staphylococcus aureus V-8 protease, the location of interchain disulfide bonds in amino (N)-terminal or carboxyl (C)-terminal fragments was determined by gel electrophoresis under reduced and nonreduced conditions . The first interchain disulfide bonds formed during dimerization are in the C-terminal region of the subunits, whereas interdimer disulfide bonds are located in the N-terminal portion . These data confirm recent electron microscopic projections of disulfide bond locations and provide support to the hypothetical role of the propolypeptide in the multimerization process. Am J Perinatol, 1987 Jan, 4(1), 59 - 62 Antibacterial activity of amniotic fluid in the early third trimester . Its association with preterm labor and delivery; Nazir MA et al.; The purpose of this study was to determine the bacterial growth inhibitory property of amniotic fluid (AF) from patients in the early third trimester and to examine its relationship to preterm labor and delivery . A total of 38 AF samples from patients between 27 and 35 weeks gestation with intact membranes were studied . Antibacterial activity was tested against Escherichia coli, Staphylococcus aureus, and Bacteroides fragilis . Fifteen fluids were from patients in preterm labor who delivered near term after successful inhibition of labor . Five of these fluids (33%) were inhibitory to E . coli, five (33%) to S . aureus, and 11 (73%) to B . fragilis . Nine fluids were from patients who failed inhibition of labor and delivered preterm; none of these were inhibitory to E . coli, two (22%) to S . aureus, and only one (11%) to B . fragilis (P less than 0.005) . Two of the nine fluids from patients who failed inhibition of labor also yielded fusobacteria . The remaining 14 fluids were obtained from patients without preterm labor and exhibited antibacterial activity similar to that from patients with successful labor inhibition . Results suggest a relationship between the absence of AF antibacterial activity against B . fragilis and preterm delivery in patients with preterm labor and intact membranes. Arch Immunol Ther Exp (Warsz), 1987, 35(6), 781 - 5 The ability of granulocytes from patients with atopy to engulf neutral latex particles and Staphylococcus aureus; Matusiewicz R et al.; The granulocytes obtained from patients with atopic diseases were investigated for their ability to engulf neutral latex particles and bacteria (S . aureus) in the presence of antigenically compatible sera of healthy subjects . Similar tests were done using the granulocytes of healthy subjects and sera of the patients with atopic diseases, and the granulocytes of these patients in autologous sera . Granulocytes of healthy subjects suspended in autologous sera served as a control . In all patients IgE globulins were determined in the serum . It was observed that only granulocytes from the patients with atopy showed a defect in engulfing latex particles and S . aureus . This defect was independent of the serum in which these granulocytes were suspended, and of the IgE level in the patient's serum. Int J Immunopharmacol, 1987, 9(6), 719 - 23 1 alpha,25-Dihydroxyvitamin D3 inhibits proliferative response of T- and B-lymphocytes in a serum-free culture; Shiozawa S et al.; The contribution of 1 alpha,25-dihydroxyvitamin D3 to the proliferative response of human B- and T-lymphocytes was examined in a serum-free culture, in which B-cells were stimulated with Staphylococcus aureus Cowan I, and T-cells with phytohemagglutinin, respectively . 1 alpha, 25-dihydroxyvitamin D3 inhibited mitogen-induced B-cell proliferation at a dose of 10(-7) M (P less than 0.01) . T-cell proliferation was inhibited at the lower dose range between 10(-9) M and 10(-7) M . Thus, although 1 alpha, 25-dihydroxyvitamin D3 acts directly on B-cells, it appears that, under physiological circumstance, 1 alpha, 25-dihydroxyvitamin D3 regulates human B-cell growth indirectly through the effect on T-cells. Cancer Detect Prev Suppl, 1987, 1, 543 - 8 B-cell reactivity in homosexuals with persistent generalized lymphadenopathy (PGL); Kekow J et al.; In addition to the well known T-cell dysfunctions in AIDS, hypergammaglobulinaemia and autoimmune phenomena indicate an involvement of the B cell as well . Reports of HTLV-III/LAV-infected B cells suggest T-cell-independent B-cell abnormalities . To look for early B-cell dysfunctions, we examined a high-risk group of AIDS consisting of six homosexuals with PGL and HTLV-III/LAV antibodies, comparing these data to those of patients with AIDS/ARC and a normal control . In vitro studies included the B-cell proliferation response (3H-thymidine uptake) to Staphylococcus aureus Cowan I and the differentiation response (Ig secretion into culture supernatants) to T-cell-dependent/-independent polyclonal B-cell activators (PBAs) . Profound alterations were found in both the proliferation and the differentiation responses . The weak response even to T-cell-independent PBAs indicates a B-cell dysfunction that is not due only to a T-cell defect in patients with PGL, similar to that observed in AIDS. Cancer Detect Prev Suppl, 1987, 1, 43 - 9 Influence of factor substitution on the B-cell response in hemophiliacs; Kekow J et al.; Studies in hemophiliacs receiving factor concentrates demonstrated T-cell defects in vitro . Recently, B-cell dysfunctions were described in AIDS and pre-AIDS and in some hemophiliacs . To investigate the B-cell function in hemophiliacs in relation to factor substitution, we examined five patients with mild (substitution less than 20,000 U/year) and seven with severe (greater than 100,000 U/year) hemophilia A and compared the data with normal control individuals . The B-cell proliferative response (3H-thymidine uptake) to Staphylococcus aureus Cowan I and the differentiation response (Ig secretion into culture supernatants) to T-cell-dependent or -independent polyclonal B-cell activators (PBAs) were studied in vitro . In contrast to T-cell dysfunctions, which correlate with the amount of clotting factor concentrates, the B-cell proliferative response was not affected . Stimulation with PBAs however failed to increase elevated spontaneous IgG levels and showed a diminished increase in IgM levels in severe, but not in mild, hemophilia . Our data give evidence of a T-cell-independent B-cell dysfunction in asymptomatic hemophiliacs that correlates with factor substitution. Toxicon, 1987, 25(6), 637 - 47 Staphylococcus aureus alpha-toxin . 2 . Reduction of epidermal growth factor receptor affinity in PC12 cells; Lazarovici P et al.; Staphylococcus aureus alpha-toxin, at sub-cytotoxic concentrations, inhibits both the 125I-labeled epidermal growth factor (EGF) binding and autophosphorylation properties of EGF-receptors in PC12 cells . This inhibition occurred only in intact cells and is probably due to a decrease in the affinity of the receptor for EGF . Streptolysin S and parcelsin could mimic the alpha-toxin effect below cytotoxic concentrations, as measured by a 51Cr release assay . In contrast, other membrane perturbing toxins with different lipid specificity, such as tetanolysin and cobra direct lytic factor, inhibited {125I}EGF binding only at cytotoxic concentrations . Staphylococcal alpha-toxin also stimulated 3-fold the specific binding of a radioactive tumor-promoting phorbol ester (PDBu) to PC12 cells at concentrations similar to those required for the inhibition of {125I}EGF binding . Although the exact mechanism for the inhibition of EGF binding by alpha-toxin has not been established, our results suggest that protein kinase C may be involved in this time-dependent process. Immunology, 1987 Jan, 60(1), 45 - 50 Activation of B chronic lymphocytic leukaemia cells by Branhamella catarrhalis; Calvert JE et al.; Cells from the blood of patients with chronic lymphocytic leukaemia were cultured in the presence of two polyclonal activators of human B cells, the bacteria Branhamella catarrhalis (Bc) and Staphylococcus aureus Cowan 1 (SAC) . Although the magnitude of the responses varied, cells from seven of the eight patients studied were induced to proliferate in response to Bc . In contrast, the response to SAC was low or negligible in seven of the eight patients, and only one patient responded well to this mitogen . Bc was also effective in inducing secretion of IgM in cells from seven of the eight patients, and this was unaffected by removal of T cells . Fractionation of CLL cells on density gradients showed that the highest level of IgM production was induced in cells with a low buoyant density, whilst cells with a high buoyant density secreted little or no immunoglobulin in response to Bc . Together, these results demonstrate that Bc is an effective, T-independent activator of both DNA synthesis and immunoglobulin production in CLL cells. Immunology, 1987 Jan, 60(1), 19 - 28 Immunocompetence of normal human appendiceal lymphoid cells: in vitro studies; Kawanishi H; In order to study the activity of immunocompetent lymphocytes in normal human adult mucosal lymphoid tissues in the gut, viable mononuclear cells (MNC) from surgically removed appendices were isolated with a rapid and improved technique, and their immune reactivity determined in vitro . Nearly two-thirds of the isolated cells were B cells, composed of sIgM (30%), sIgA (19%) and sIgG (12%), less than one-third of cells being T cells, composed of OKT4+ helper/inducer T cells (18%) and OKT8+ suppressor/cytotoxic T cells (9%) . Isolated B cells were responsive to SAC (Cowan type I Staphylococcus aureus) with marked augmentation of IgG production, but not to lipopolysaccharides (LPS) . In the presence or absence of pokeweed mitogen (PWM) and protein A, all isotype-specific Ig production was augmented along with increasing ratios of autologous T cells to B cells, whereas in the presence of anti-HLA-DR antibodies the production of IgA and IgM was reduced to a great extent . Finally, in vitro induction of concanavalin A (Con A)-induced suppressor T (Ts) cells was less than that of helper T (Th) cells . Thus, normal adult appendiceal (Ap) lymphocyte reactivity is predominated by helper T (Th) cells; IgA isotype-specific Ig production appears to be largely regulated by the autoreactive Th subset . This responsiveness to exogenous stimuli may play an important role in the gut mucosal immune responsiveness. Arch Intern Med, 1987 Jan, 147(1), 89 - 93 B-cell activation and immunoregulation in end-stage renal disease patients receiving hemodialysis; Raskova J et al.; B-lymphocyte functions were studied in peripheral blood mononuclear cells of end-stage renal disease patients undergoing intermittent hemodialysis for longer than two years . T-cell-dependent B lymphocyte proliferation after pokeweed mitogen stimulation was low in half of the hemodialyzed patients . T cell-independent B cell response to Staphylococcus aureus, Cowan I, was also significantly reduced . Spontaneous production of immunoglobulin in cultures of peripheral blood mononuclear cells of uremic patients was comparable with that of healthy controls, but pokeweed mitogen-stimulated antibody secretion was significantly reduced with cells from patients undergoing hemodialysis . Helper T-cell functions in B-cell activation were also qualitatively deficient in uremic patients . It is concluded that B-cell activation and immunoregulation is defective in patients undergoing long-term hemodialysis. Arch Immunol Ther Exp (Warsz), 1987, 35(2), 169 - 73 The influence of synthetic tuftsin and its analogs on the function of granulocytes of children with acute lymphoblastic leukemia; Kazanowska B et al.; The effect of tuftsin, its synthetic analogs and arginine on phagocytosis of Staphylococcus aureus by granulocytes from leukemic children was investigated in vitro . The high stimulatory effect of tuftsin and arginine was shown . The decrease of phagocytosis of PMN from healthy subjects after preincubation with its analogs and arginine was observed, suggesting the regulatory effect of these peptides. Drugs, 1987, 34 Suppl 1, 124 - 30 Ofloxacin treatment in the management of chronic osteitis; Ketterl R et al.; In an open prospective study, the efficacy and tolerability of ofloxacin in the treatment of chronic post-traumatic osteitis was examined in 83 patients . 103 different pathogens were isolated . More than 75% were Gram-positive bacteria with Staphylococcus aureus occurring in 61% of cases . After ofloxacin treatment bacteriological elimination was more than 90% for both Gram-positive and Gram-negative species, leading to a clinical cure in 85% of patients . Reinfection occurred in 5% of patients . These values were obtained from follow-up examinations which were carried out at least 6 months after the end of therapy . The tolerability of ofloxacin was excellent and no drug-related allergic reactions or side effects were observed . In conjunction with adequate surgical treatment, ofloxacin proved to be a useful antibacterial agent in the therapy of chronic bone infection. J Leukoc Biol, 1987 Jan, 41(1), 78 - 82 Effect of alkylating agents on the clearance of Staphylococcus aureus from murine lungs; Nugent KM et al.; Alkylating agents have several effects on cellular host defense responses which could increase either the frequency or the severity of pulmonary infections . In addition, some of these agents directly injure lung parenchyma and could have effects on intrapulmonary killing processes independent of any effect on phagocyte number and function . We have used a murine model for staphylococcal clearance to evaluate the effect of cyclophosphamide and mechlorethamine on intrinsic lung defenses . Single doses of mechlorethamine (40 micrograms IV) or of cyclophosphamide (150 mg/kg IP) reduce peripheral blood neutrophil counts and spleen weights on day 3 after injection; with the exception of neutrophil counts in mechlorethamine-treated mice, these parameters returned to normal by days 10-12 . Both drugs reduced the number of alveolar macrophages recoverable by bronchoalveolar lavage on days 10-12 but not day 3 . Mechlorethamine delayed the clearance of Staphylococcus aureus 502A from the lung on both days 3 and 12, but cyclophosphamide did not alter clearance on either day 3 or 10 . The defect in clearance in mechlorenthamine-treated mice resolved by 3 wk after drug administration . These results demonstrate that alkylating agents do not have uniform effects on antibacterial processes in the murine lung . Since the mechlorethamine effect on staphylococcal elimination appears independent of its effect on macrophage numbers, these results suggest that staphylococcal clearance also depends on nonphagocytic host defense factors. Clin Ther, 1987, 10(1), 82 - 91 Parenteral antibiotic therapy at home: experience with intramuscular cefonicid; Nadworny HA et al.; Patients with serious infections often remain hospitalized solely to continue parenteral antibiotic therapy although many who are afebrile and medically stable could complete treatment in an outpatient setting . Potential advantages of outpatient treatment include lower costs and greater efficiency in the use of hospital beds, and economic and psychological benefits to patients . Cefonicid, a parenteral cephalosporin with an extended antibacterial spectrum and a prolonged serum half-life, was used to treat 39 patients with serious infections, mostly those of bone or soft tissues . Staphylococcus aureus was the predominant pathogen isolated (18 patients); nine gram-negative aerobes were observed in single instances . After indicated surgical procedures and inpatient intravenous antibiotic therapy, the patients were discharged to continue parenteral therapy at home with once-daily intramuscular doses of cefonicid . Mean duration of outpatient therapy was 16 days . The clinical response was satisfactory in all but two patients, one with relapse of a S aureus wound infection, the second with a gram-negative reinfection at the site of a bone cyst . Systemic and local tolerability of cefonicid were good . It is concluded that outpatient antibiotic therapy with intramuscular cefonicid can benefit both patients and hospitals. Trans R Soc Trop Med Hyg, 1987, 81(5), 829 - 32 Replication of Japanese encephalitis virus in monkey, pig and chick leucocyte cultures; Kedarnath N et al.; Japanese encephalitis (JE) virus replicated in monkey, pig and day-old chick leucocyte cultures . The titres obtained on days 3 to 5 after infection in monkey, pig and chick leucocyte cultures were comparable . Treatment of monkey leucocyte cultures with the mitogens phytohaemagglutinin P, pokeweed mitogen (PWM), formalinized Staphylococcus aureus (Cowan I) or concanavalin A and pig leucocytes with PWM did not significantly affect their ability to support replication of JE virus . No relationship was observed between the amount of {3H}thymidine incorporated in untreated or mitogen treated monkey or pig leucocyte cultures and the titres of JE virus in such cultures . The ability of monkey, pig and chick leucocyte cultures to support JE virus replication was abrogated following silica treatment . These findings suggest that monocytes may serve as one of the important sites of JE virus replication. Reprod Nutr Dev, 1987, 27(6), 999 - 1003 Effects of intrauterine infection by Staphylococcus aureus and Mycoplasma capricolum on the fertility of Nubian goats; Shallali AA et al.; Oestrous cycles were followed by blood progesterone titration and the occurrence of pregnancy was observed after mating in three groups of mature female Nubian goats . Two groups of 4 and 3 goats each were experimentally infected by an in utero injection of either a Staphylococcus aureus or a Mycoplasma capricolum strain of caprine origin . A third group of 4 goats served as a control . Both bacterial infections persisted in all experimental animals, except one, for at least 70 days . As a result, there is a shortening of the first two cycles and progesterone concentration was lower than in the controls . Only one inoculated goat became pregnant after the third oestrus . It is suggested that bacterial inflammation of the uterus may thus modify the oestrous cycle through the prostaglandin pathway. Magnesium, 1987, 6(6), 325 - 9 Hypomagnesemia, hypocalcemia, and toxic-shock syndrome . A case report; Rudick JH et al.; Hypocalcemia is common in toxic-shock syndrome (TSS); however, the role of magnesium deficiency in TSS remains to be defined . A previously healthy nurse on no maintenance medication developed hypocalcemia and hypomagnesemia in association with characteristic TSS, presenting with fever, headache, mental confusion, erythroderma, watery diarrhea and abnormal liver functions tests . Coagulase-positive Staphylococcus aureus as well as staphylococcal toxin were isolated from the cervix . Calcitonin levels were normal . Serum magnesium and calcium levels were low at presentation and later intravenous magnesium loading demonstrated a marked rise in 1,25-(OH)2 vitamin D and parathormone (PTH), with high retention of the infused load consistent with functional hypoparathyroidism . Intracellular magnesium deficiency may be a significant factor in the pathogenesis of hypocalcemia in TSS and warrants routine clinical consideration. Scand J Infect Dis, 1987, 19(6), 623 - 7 Bacteremic Staphylococcus aureus pneumonia; Watanakunakorn C; Medical records of 44 consecutive patients with bacteremic Staphylococcus aureus pneumonia between 1980 to 1984 in a 750-bed community teaching hospital were reviewed . 36 patients (82%) were 60 years or older and the median age was in the 8th decade . There was no seasonal variation in the prevalence . The infection was hospital-acquired in 29 patients (66%) . All patients had 1 or more (median 3) major underlying conditions . The chest roentgenogram was non-diagnostic; only 1 patient had empyema . Fever and leukocytosis were common . Many patients had renal insufficiency and hyperbilirubinemia . The overall mortality was 84%. Acta Microbiol Hung, 1987, 34(2), 139 - 45 Protective activity of two human intravenous immunoglobulin preparations in experimental infection with an encapsulated Staphylococcus aureus strain; Vassilev T et al.; The protective effect of two different human immunoglobulin preparations for intravenous use and one specific staphylococcal immunoglobulin for intramuscular application were compared in mice infected with the capsular Staphylococcus aureus Smith strain . Immunovenin is produced by partial fragmentation of IgG with plasmin; it contains about 60% intact IgG and 40% Fab and Fc fragments . Immunovenin-intact is produced by a polyethylene glycol (mol wt 6000) fractionation method followed by ion exchange chromatography and contains more than 95% intact IgG molecules . A specific staphylococcal immunoglobulin is obtained by a rivanol/ethanol fractionation method from selected sera with high anti-alpha toxin level . All three types of immunoglobulins induced similar degree of protection when their effect was determined as activity (ED50) per gram immunoglobulin. Ter Arkh, 1987, 59(10), 57 - 61 {Invasive treatment methods in therapy and angiogenic sepsis}; Gogin EE et al.; In 1850 patients with intravenous catheters, intravenous shunts and fistulas sepsis developed in 0.43% . Infection of the catheters and shunts was more frequent than sepsis development . The growth of microorganisms (Staphylococcus aureus, Gram-negative microbes) was noted during a bacteriological study of 300 removed catheters in 27.7% . Angiogenic sepsis was caused by Staphylococcus aureus . Peculiarities of the clinical picture of angiogenic sepsis were discussed . Its early diagnostic signs were described . Immunological changes in the patients with angiogenic sepsis were evaluated . The problems of pathogenesis and prevention of angiogenic sepsis were given due attention. Pediatr Cardiol, 1987, 8(4), 275 - 7 False aneurysm of the right ventricle due to endocarditis in a child; Garty BZ et al.; An eight-year-old boy with supravalvular pulmonic stenosis, supravalvular aortic stenosis, and ventricular septal defect developed Staphylococcus aureus endocarditis . The infection was complicated by formation of a false aneurysm of the right ventricular outflow tract, which was demonstrated by contrast echocardiogram . Surgical treatment was successful . This is a unique case of false aneurysm of the outflow tract of the right ventricle, because it is secondary to endocarditis without known previous trauma to the right ventricular wall. Zentralbl Mikrobiol, 1987, 142(5), 379 - 86 Status of membrane-filter procedure in the examination of water and milk for Staphylococcus aureus; Sabbour MM et al.; A comparative study was carried out to assess the performance of four selective media, namely Baird-Parker (B-P), Mannitol-Salt Agar (M.S.A.), Staphylococcus-110 (S-110), and Chapman-Stone (C.S) in solid and liquid forms for recovery of Staphylococcus aureus from water and milk, using membrane filter technique . The descending order of efficiency for demonstrating the presence of inoculated S . aureus from unsterilized Nile water was (B-P)--(M.S.A) + (S-110) and (C.S) medium in solid form and changed to (S-110)--(M.S.A)--(C.S) when used in the liquid form . With sterile Nile water, the descending order of efficiency of the tested media was (B-P) + (M.S.A)--(S-110) and (C.S) medium in the solid form and (M.S.A) + (S-110) and (C.S) in the liquid form . In testing the inoculated milk using solid media, (S-110) and (M.S.A) tend to show similar results and took intermediate position between (B-P) and (C.S) . When liquid media were used, the descending order of productivity was (S-110)--(M.S.A) and (C.S). Med Cutan Ibero Lat Am, 1987, 15(2), 140 - 4 {Angiolymphoid hyperplasia with eosinophilia . Apropos of a case of peculiar development: importance of its correct diagnosis and treatment}; Gatti JC et al.; We present the case of a patient thirty seven years old who has angiolymphoid hyperplasia with eosinophilia, diagnosis confirmed by histopathological examination, localized in ear's pavilion, ear's outer duct and beyond the ear . She has received intralesional injections of corticoid (because of a possible misdiagnosis) . This provoked the appearance of a pyogenous process with a profuse, purulent and fetid secretion (Staphylococcus aureus) resistant to many antibiotics which was finally controlled with trimethoprim-sulfamethoxazole . We controlled the disease with radiotherapy applications and afterwards partial cryosurgery. Prog Clin Biol Res, 1987, 240, 3 - 19 Staphylococcus aureus V8-protease catalyzed segment exchange reaction of alpha-chain of hemoglobin S: a semisynthetic approach for the preparation of variants of alpha-chain; Acharya AS et al.; The Glu(30)-Arg(31) peptide bond of alpha-chain of hemoglobin is readily and quantitatively hydrolyzed by Staphylococcus Aureus V8-protease at pH 4.0 and 37 degrees C . This region of the alpha-chain represents a 'permissible discontinuity region' of the chain within its tertiary interactions . Protease catalyzed reformation of peptide bonds in such permissible discontinuity regions of fragment complementing systems of proteins appears to be a useful procedure for the preparation of variants of the protein . Therefore, attempts have been now made to 'restitch' the segments alpha 1-30 and alpha 31-141 using V8-protease . The alpha-amino group of alpha 1-30 was selectively dihydroxypropylated (DHP) using {14C}-glyceraldehyde to follow the protease catalyzed reformation of globin . V8-protease catalyzed the condensation of N alpha-DHP-alpha 1-30 and alpha 31-141 to generate N alpha-DHP-alpha 1 141 in the presence of 30% propanol at pH 6.0 and 4 degrees C . The synthetic yield of N alpha-DHP-alpha 1-141 is about 55% in 48 hrs and remained nearly the same even after seven days . Under the same conditions alpha-globin was digested at the Glu(30)-Arg(31) peptide bond to nearly 40% . Thus, the amount of Glu(30)-Arg(31) peptide bond formed in 48 hrs appears to be the result of an equilibrium in the V8-protease catalyzed hydrolytic and synthetic reactions . The reformation of Glu(30)-Arg(31) bond appears to be very selective . V8-protease did not catalyze the peptide bond formation in an equimolar mixture of alpha 31-141 with either alpha 1-23, alpha 1-27, or alpha 28-30 all of which have glutamic acid as the carboxy terminal residue just as in alpha 1-30 . The high selectivity in the protease catalyzed formation of Glu(30)-Arg(31) peptide bond suggests that the segment alpha 1-30 of the intact alpha-globin could be exchanged with synthetic analogs of alpha 1-30 using V8-protease without actually isolating the fragment alpha 31-141 . Incubation of {14C}-N alpha-DHP-alpha 1-30 with alpha-globin at pH 6.0 and 4 degrees C along with V8-protease in 30% n-propanol indeed resulted in the exchange of the segment alpha 1-30 of alpha-globin with {14C}-N alpha-DHP-alpha 1-30 . This V8-protease catalyzed 'segment exchange reaction' should facilitate the preparation of alpha-chains with double mutations from the large number of naturally occurring alpha-chains with single mutation.(ABSTRACT TRUNCATED AT 400 WORDS) Microbiol Immunol, 1987, 31(7), 649 - 56 Induction of resistance in mice by the capsular polysaccharide antigens of Staphylococcus aureus; Yoshida K et al.; Mice actively immunized with capsular polysaccharides extracted from capsular type strains A, B, C, and D, determined by the serum-soft agar technique, were protected against lethal infection by homologous strains, but no animals survived infection by heterologous substance immunization even with at high doses . Passive protective antibody in rabbit antisera prepared using these strains was absorbed out only by homologous capsular polysaccharide in mice . These results indicated that resistance was specific for capsular polysaccharide . The substance contained mainly neutral sugar, small amounts of hexosamine, methyl-pentose, and phosphate although these amounts varied depending on the capsular types strains. C R Acad Sci III, 1987, 305(17), 623 - 6 {Expression, export and one-step purification of proteins by fusion to the MalE protein of E . coli}; Bedouelle H et al.; Enzymes can be fused at the C-terminal end of the maltose binding protein (MalE), at the genetic level . Expression of the hybrid proteins, under control of promoter malEp and of the constitutive activator, MalTc1, can be repressed by glucose . The hybrid proteins are localised either in the bacterial cytoplasm or periplasmic space, depending on whether MalE harbors a signal peptide mutation or not; as MalE, they can be purified in one step by chromatography on cross-linked amylose . The Staphylococcus aureus Nuclease and the Klenow portion of E . coli DNA-polymerase I keep their specific activities when fused to MalE. Acta Med Scand, 1987, 222(5), 465 - 70 Staphylococcus aureus bacteremia in patients with hematological malignancies and/or agranulocytosis; Espersen F et al.; A total of 6,253 cases of Staphylococcus aureus bacteremia, including 274 (4.4%) endocarditis cases, were registered in Denmark in the period 1975-1984 . Patients with hematological malignancies and/or agranulocytosis accounted for 479 of the bacteremia cases . The incidence of endocarditis in this group of patients was only 0.4% as compared to 4.7% in other patients with staphylococcal bacteremia (p less than 0.01) . The lower incidence of endocarditis complicating bacteremia in these patients may justify a shorter course of therapy than usually recommended for suspected endocarditis . Patients with hematological malignancies and other patients with agranulocytosis had a higher mortality (49 and 46%, respectively) than other patients with S . aureus bacteremia (33%) . The highest mortality was found in patients with multiple myeloma (71%, p less than 0.01), the lowest in patients with acute lymphocytic leukemia (28%, p less than 0.01) . The higher mortality in these patients may indicate that empiric antibiotic regimens in granulocytopenic patients should include a specific anti-staphylococcal agent. Infect Control, 1987 Jan, 8(1), 7 - 14 Prospective microbiologic surveillance in control of nosocomial methicillin-resistant Staphylococcus aureus; Walsh TJ et al.; A prospective microbiological surveillance (PMS) program was developed in a comprehensive hospital-wide effort for control of nosocomial methicillin-resistant Staphylococcus aureus (MRSA) . This PMS program entailed: active identification of colonized and infected patients; application of a screening microbiologic system for MRSA; isolation of colonized and infected patients; antibiotic decolonization of MRSA; and educational efforts . The PMS program was studied over three and one half years for its contribution to infection control of MRSA, early identification of nosocomial MRSA outbreaks, use of the highest yield surveillance culture sites, and cost effectiveness . Following initiation of the PMS program in December 1982, during an MRSA outbreak, the frequency of new MRSA cases declined from 14 to none by the end of a 3-month pilot study . The frequency of new MRSA cases stabilized at approximately 2 per month until October 1983, when the PMS system allowed prompt detection of a new outbreak of 11 cases . Following isolation and antibiotic decolonization, the frequency of cases again declined to 3 per month . A third outbreak in December 1985 again was promptly detected and controlled . Infection to colonization ratio decreased from a maximum of 1.5 during outbreaks to a minimum of 0.17 after outbreaks . Wounds and tracheostomy sites provided the greatest yield of detection of new cases of MRSA . During one 15-month period, 35 of the 43 new cases were detected initially at wounds and tracheostomy sites . No new MRSA cases were detected by a positive axillary or nares site alone . The estimated quarterly cost of outbreaks and infection paralleled the quarterly frequency of new MRSA cases.(ABSTRACT TRUNCATED AT 250 WORDS) Immunology, 1987 Jan, 60(1), 51 - 5 Enhancement of the B-cell response to Staphylococcus aureus Cowan strain 1 by natural human gamma interferon; Aoki N et al.; The effects of interferon (IFN) on the B-cell response to Staphylococcus aureus Cowan strain 1 (SAC) were studied comparatively with natural human IFN-alpha, IFN-beta, and IFN-gamma, employing equal units of their anti-viral activity . First, the response was investigated in peripheral mononuclear cells obtained from healthy individuals, and next, confirmed in cultures employing B-cell enriched populations derived from tonsils obtained at tonsillectomy from patients with chronic tonsillitis . B cells were purified by rosetting out T cells with sheep red cells followed by the removal of adherent cells on a plastic surface . The results show that the SAC-stimulated lymphoproliferative response was enhanced in the presence of IFN-gamma in a dose-related manner, at concentrations ranging from 10 to 1000 IU/ml, both in peripheral mononuclear cells and tonsillar B-cell enriched fractions . In contrast, IFN-alpha and IFN-beta did not enhance or suppress SAC-stimulated blastogenesis in either lymphocyte preparation . The enhancing effects specific to IFN-gamma were more remarkable in cultures stimulated with a suboptimal dose (0.002%) of SAC than when the optimal dose (0.005%) was employed. J Clin Microbiol, 1987 Jan, 25(1), 122 - 7 Antibody response to teichoic acid and peptidoglycan in Staphylococcus aureus osteomyelitis; Jacob E et al.; An enzyme-linked immunosorbent assay was used to evaluate the immunoglobulin G (IgG) response to Staphylococcus aureus crude teichoic acid (TA) and peptidoglycan (PG) in both rabbits and patients with osteomyelitis . In rabbits with experimental S . aureus osteomyelitis, elevated levels of IgG to TA were present in 13/18 (72%) of the serum samples obtained at 4 and 10 weeks postinfection . In contrast, only 5/18 (28%) of these sera were found to be positive for antibodies to PG . Of a total of 39 patients with confirmed S . aureus osteomyelitis (11 acute, 28 chronic), IgG to TA was elevated in 17 (44%), whereas antibodies to PG were found to be increased in only 1 (3%) . Cross-reacting antibodies to S . aureus TA were detected in only 1/18 (6%) of the patients with osteomyelitis caused by organisms other than S . aureus . These studies indicate that IgG to TA is more prevalent than IgG to PG in patients with staphylococcal osteomyelitis . Although these results are encouraging, a larger number of patients is required for an adequate evaluation of the TA enzyme-linked immunosorbent assay for the diagnosis and management of suspected S . aureus osteomyelitis. Int J Pept Protein Res, 1987 Jan, 29(1), 21 - 39 Amino acid sequence of UP1, an hnRNP-derived single-stranded nucleic acid binding protein from calf thymus; Merrill BM et al.; The UP1 single-stranded nucleic acid binding protein from calf thymus (Herrick, G . & Alberts, B.M . (1976) J . Biol . Chem . 251, 2124-2132) has recently been shown to be a proteolytic fragment derived from the A1 heterogeneous nuclear ribonucleoprotein (hnRNP) (Pandolfo et al . (1985) Nucleic Acids Res . 13, 6577-6590) . The NH2-terminus of the 22,162 dalton UP1 protein appears to be blocked, which suggests that UP1 represents the NH2-terminal two thirds of this 32,000 dalton hnRNP protein . The complete amino acid sequence for UP1 was derived from automated sequencing of peptides that were purified by HPLC from digests with trypsin, chymotrypsin, Staphylococcus aureus protease, endoproteinase Lys-C, and cyanogen bromide . Trichloroacetic acid precipitation followed by enzymatic digestion in 2 M urea proved to be the best approach for generating UP1 peptides . By carboxymethylating after, rather than before, digestion it was possible to avoid problems associated with the insolubility of the carboxymethylated UP1 . All of the resulting peptides in amounts varying from 2 to 15 nmol were coupled to aminopolystyrene prior to solid-phase sequencing . Using these methods, no difficulties were encountered in assigning glutamic acid residues or in completely sequencing peptides that contained up to 25-30 residues . The relative ease with which the UP1 protein was sequenced, requiring only about a year to complete, and the comparatively modest amount of protein required, less than 5 mg, attests to the usefulness of water soluble carbodiimide coupling and solid-phase sequencing for determining the primary structures of proteins . In addition to serving as a basis for determining structural relationships among various mammalian single-stranded nucleic acid binding proteins, the amino acid sequence of UP1 reveals that the A1 hnRNP protein contains a region of internal sequence homology that apparently corresponds to two independent nucleic acid binding sites. J Virol Methods, 1987 Jan, 15(1), 33 - 9 Detection of IgA antibody to EBV membrane antigen using Staphylococcus aureus preabsorbed sera is closely associated with nasopharyngeal carcinoma; Pi GH et al.; The presence of IgA antibody to membrane antigen (MA) of Epstein-Barr virus (EBV) was tested in sera from 48 nasopharyngeal carcinoma (NPC) patients, 40 patients with tumors other than NPC and 46 normal individuals . The sera were preabsorbed with Staphylococcus aureus (SPA) (strain no . 1800) prior to their use in the indirect immunofluorescence test . One hundred percent of the NPC patients had the IgA/MA antibody with a GMT of 1:141 . In patients with tumors other than NPC or normal individuals, IgA/MA antibodies were not detectable . The IgA/MA antibodies have been demonstrated in 6 NPC patients lacking detectable antibody levels in the indirect immunofluorescence test using nonabsorbed sera . Our data indicate that preabsorbtion of sera with SPA renders the diagnostic test significantly more sensitive for the detection of the nas