Staphylococcus aureus

J Dairy Sci, 1988 Nov, 71(11), 3112 - 9
Bovine vitamin A and beta-carotene intake and lactational status . 1 . Responsiveness of peripheral blood polymorphonuclear leukocytes to vitamin A and beta-carotene challenge in vitro; Tjoelker LW et al.; Dietary vitamin A and beta-carotene were assessed on their interaction with lactational status to influence neutrophil function in vitro . Cows were fed 1) 53,000 IU or 2) 213,000 IU vitamin A, or 3) 53,000 IU vitamin A plus 400 mg beta-carotene/cow per d from 6 wk before to 2 wk after dry off . Blood neutrophils were isolated the day of dry off and 2 wk after dry off and incubated with retinol, retinoic acid, or beta-carotene . Phagocytosis and kill of Staphylococcus aureus were measured . Across all treatments, kill was higher after dry off than before dry off . Phagocytosis tended to be lower after dry off than before in cows fed vitamin A only . In vitro, 10(-6) M beta-carotene stimulated phagocytosis after dry off and kill before dry off in cows fed vitamin A only . In general, retinol and retinoic acid suppressed phagocytosis but did not affect kill . Neutrophils from cows fed high amounts of vitamin A were more susceptible to in vitro suppression than those from cows fed adequate amounts of vitamin A . Therefore, vitamin A and beta-carotene supplementation interacts with lactational status to influence the responsiveness of bovine neutrophils to vitamin challenge in vitro.

Zentralbl Bakteriol Mikrobiol Hyg {A}, 1988 Nov, 270(1-2), 115 - 21
A new chromogenic assay for direct detection of staphylocoagulase; Bulanda M et al.; Tube test for detection of staphylococcal coagulase, despite of many disadvantages, is commonly used in clinical microbiology for identification of Staphylococcus aureus . In this paper a new chromogenic method for detection of the coagulase directly in staphylococcal cultures is described and evaluated on the basis of a comparison with the standard tube assay . The chromogenic assay appeared to be as sensitive as the tube test but results of the former one can be read in a few hours without any apparatus.

J Burn Care Rehabil, 1988 Nov-Dec, 9(6), 613 - 5
Use of nonsterile gloves for routine noninvasive procedures in thermally injured patients; Sadowski DA et al.; A total of 26 boxes of gloves were analyzed to determine if using nonsterile gloves for routine noninvasive procedures was sufficient for thermally injured patients and if the risk of infection increased . All of the study boxes had some organism present on or in the used box; the most common type found was Staphylococcus aureus . Eleven of the 13 subjects (85%) had specific antibiotic-resistant strains of S . aureus present on cultures obtained from open wounds . Seven (64%) of these corresponded to the glove boxes assigned to that patient . The remaining four boxes of gloves had no S . aureus present . In all of the boxes of gloves that had positive S . aureus cultures, 100% of the resistant strains occurred after it was first cultured from the patient . As a result, nonsterile gloves can be used safely for routine non-invasive procedures in the thermally injured patient . It is imperative to avoid using a common box of gloves for two or more patients to prevent the transfer of organisms between patients.

Zentralbl Bakteriol Mikrobiol Hyg {A}, 1988 Nov, 269(3), 387 - 94
{In vitro bactericidal effects of ofloxacin on S . aureus}; Korting HC et al.; Two Staphylococcus aureus (S . aureus) strains with differing susceptibility to penicillin G as expressed by the minimum inhibitory concentration were exposed in vitro over a period of 8 h to the continuously changing concentrations of ofloxacin achieved in human serum and cantharides blister fluid (CBF) after the single oral application of 600 mg . In both strains bacterial density was rapidly, but not totally reduced: a reduction by 99 percent took no longer than about 1 and 1.5 h resp . facing the serum level profile and about 1.5 h facing the tissue level profile . The maximum reduction of staphylococcal density in percent (kn) amounted to 0.0007 and 0.0034 and to 0.0038 and 0.0047 resp . Thus not only the serum level profiles but also the skin tissue level profiles obtainable in man proved highly effective in vitro against S . aureus irrespective of the bacterial suspectibility against penicillin G . Therefore oral ofloxacin should prove useful in staphylococcal diseases especially in so far as cutaneous tissue is involved.

Med Hypotheses, 1988 Nov, 27(3), 215 - 6
The role of oxygen in toxic shock syndrome; Mahoney JR Jr; The etiology of toxic shock syndrome (TSS) has been extensively investigated in recent years . It is generally accepted that the causative organism is Staphylococcus aureus . Certain strains of this bacterium produce one or more toxins which are thought to be responsible for the symptoms associated with TSS . There is no general agreement, however, as to why menstruating women who use tampons are of particular risk for developing this disease . More recently, TSS has also been associated with the use of absorbent contraceptive sponges . In this paper I will present evidence that the introduction of oxygen into the vaginal cavity during tampon insertion is responsible in part for the development of TSS . Furthermore, I will discuss several methods for the safe removal of this exogenous oxygen.

J Antibiot (Tokyo), 1988 Nov, 41(11), 1602 - 16
Synthesis and biological activity of a new cephalosporin, BMY-28232 and its prodrug-type esters for oral use; Kamachi H et al.; The synthesis and structure-activity relationships of 7-{(Z)-2-(2-aminothiazol-4-yl)-2-hydroxyiminoacetamido}-3-{( Z)-1- propenyl}-3-cephem-4-carboxylic acid (BMY-28232), its 3-alkenyl analogs (6 and 7) and O-substituted derivatives of the oxyimino moiety (10) are described, as well as the oral pharmacokinetics and in vivo activities of the 1-acetoxyethyl ester of BMY-28232 (BMY-28271) and its analogous esters (11) . The 3-alkenyl groups were introduced by the Wittig reaction of the ylide (2) prepared from the 3-chloromethyl cephem (1) to afford the Z (main) and E (minor) isomers regarding the 3-side chain . The O-substituted derivatives (10) were prepared by 7-N-acylation of the 7-amino cephem (4a) with the corresponding O-substituted side chain acids (8) . The prodrug esters (11) were prepared by esterification of BMY-28232 with an appropriate halide . BMY-28232 was the most active among the 3-alkenyl analogs tested against Gram-negative organisms and much more active than the O-substituted derivatives against Gram-positive bacteria . BMY-28271 showed good oral bioavailability (66%) and good in vivo efficacy in mice against infections of Staphylococcus aureus Smith (PD50, 0.68 mg/kg) and Escherichia coli Juhl (0.54 mg/kg).

Contraception, 1988 Nov, 38(5), 573 - 8
Humoral immunity in oral contraceptive users . II . In vitro immunoglobulin production; Bisset LR et al.; The worldwide acceptance of steroid-based oral contraception makes it imperative that the effect of these agents on the immune system is understood . Nevertheless, information regarding the effect of steroid-based oral contraception on humoral immunoregulation is limited . In this report the in vitro production of IgG and IgM is measured following stimulation with either the T-dependent activator pokeweed mitogen (PWM) or the T-independent activator fixed/killed Staphylococcus aureus Cowan I (StaCw) . No significant differences are observed between the in vitro IgG or IgM levels following stimulation with PWM or StaCw for females taking steroid-based oral contraceptives and females not taking steroid-based oral contraceptives . We conclude that humoral immunoregulation is unaltered in steroid-based oral contraceptive usersPIP: Production of IgG and IgM immunoglobulins in vitro by purified monocytes isolated from the blood of women taking oral contraceptives was assayed by the sensitive ELISA method . The subjects were taking pills containing ethinyl estradiol and levonorgestrel . Washed mononuclear cell suspensions containing no more than 5% polymorphonuclear cells were incubated 7 days with either formaldehyde-killed Staphylococcus aureus strain Cowan I or T-cell dependent pokeweed mitogen . No significant differences were observed in immunoglobulin production between pill users and controls .

Am J Med, 1988 Nov, 85(5), 615 - 8
Clinical significance and pathogenesis of hyperbilirubinemia associated with Staphylococcus aureus septicemia; Quale JM et al.; PURPOSE: Our goal was to examine the clinical significance of hyperbilirubinemia in patients with Staphylococcus aureus endocarditis . In addition, preliminary data concerning the possible mechanism of cholestasis observed during S . aureus septicemia are presented . PATIENTS AND METHODS: This study had two parts: a clinical investigation and a laboratory investigation . In the former, patients with endocarditis were identified through chart review . Those with admission total serum bilirubin levels of 2.0 mg/dl or greater were considered to have hyperbilirubinemia . In the latter investigation, the hepatic storage capacity and transport maximum for sulfobromophthalein (BSP), an organic dye that is rapidly taken up and excreted by the liver, were determined by measuring the change in serum concentration and the corresponding hepatic removal rate at various BSP infusion rates . Measurements were conducted before and after the infusion of Escherichia coli-derived lipopolysaccharide in some rabbits, after the infusion of resuspended S . aureus in others, and after the infusion of lipoteichoic acid in the remainder . RESULTS: Eleven of 47 consecutive patients with S . aureus endocarditis were noted to have hyperbilirubinemia without clinical or laboratory evidence of hepatic bacterial infection . Compared with the remaining 36 patients, these 11 patients had a significantly lower mean platelet count and a higher serum creatinine level and white blood cell count . Although none of the 47 patients were hypotensive on admission, four of the 11 hyperbilirubinemic patients died of overwhelming sepsis, compared with two of the 36 remaining patients (p less than 0.05) . When one of the clinical isolates of S . aureus or lipoteichoic acid was infused into conscious rabbits, there was a marked decrease in the hepatic transport maximum and an increase in the relative hepatic storage capacity of sulfobromophthalein . Similar changes were noted following the administration of lipopolysaccharide . CONCLUSION: Our data suggest that the presence of hyperbilirubinemia in patients with S . aureus sepsis may identify persons at high risk of dying from overwhelming sepsis . It further suggests that lipoteichoic acid may play an important role in causing defective hepatic excretory function that is responsible for hyperbilirubinemia.

J Pediatr, 1988 Nov, 113(5), 826 - 30
Role of respiratory syncytial virus in early hospitalizations for respiratory distress of young infants with cystic fibrosis; Abman SH et al.; To determine the frequency of respiratory syncytial virus (RSV) as the cause of hospitalization for acute pulmonary exacerbations in young infants with cystic fibrosis (CF), and to assess the clinical effects of RSV infections, we prospectively followed 48 children with a diagnosis of CF after identification by newborn screening . At a mean follow-up age of 28.8 months (range 5 to 59), 18 infants (38%) had been hospitalized a total of 30 times for acute respiratory distress . At the time of admission, 18 infants (60%) were less than 12 months, 8 (27%) between 12 and 24 months, and 4 more than 2 years of age . The RSV was identified in seven hospitalized infants, as determined by fluorescent antibody, immunoassay, or culture . Before admission with RSV infection, one of the seven infants had chronic respiratory signs, none had Brasfield chest x-ray scores below 20, and a previous throat culture was positive for Staphylococcus aureus in one infant . Hospitalizations were prolonged (mean duration 22 days), and were characterized by significant morbidity, with three infants (43%) requiring mechanical ventilation and five infants (71%) requiring home oxygen therapy for persistent hypoxemia at discharge . At a mean follow-up age of 26 months, these infants more frequently have chronic respiratory signs (p less than 0.01) and lower chest radiograph scores (p less than 0.05) than other CF infants . These findings demonstrate that RSV is an important cause of early acute respiratory tract morbidity in young infants with CF, and suggest the need for studying new strategies to implement early and aggressive antiviral therapy in young infants with CF.

Chest, 1988 Nov, 94(5), 1088 - 90
Osler's nodes on the dorsum of the foot; Watanakunakorn C; Osler's nodes developed on the dorsum of the left foot of a patient with enterococcal endocarditis and on the dorsum of the right foot of another patient with Staphylococcus aureus endocarditis . The lesions were erythematous, raised, and very painful, but they resolved completely . There was no evidence of other embolic phenomena at the time the Osler's node appeared.

Plasmid, 1988 Nov, 20(3), 271 - 5
Nucleotide sequence of a staphylococcal plasmid gene, vgb, encoding a hydrolase inactivating the B components of virginiamycin-like antibiotics; Allignet J et al.; The nucleotide sequence of a 1883 bp fragment isolated from a resistance plasmid harbored by a Staphylococcus aureus clinical isolate and carrying the gene, vgb, encoding a hydrolase inactivating the B components of virginiamycin family has been determined . The sequence contains one open reading frame which extends from the ATG codon at nt 641 to a TGA codon at nt 1537 and which potentially codes for a protein of 33.035 Da . This value is in agreement with the apparent size (33 kDa) of the protein observed, in minicell extracts . Inactivation of the B components of the virginiamycin antibiotics as well as resistance to these antibiotics were expressed in a virginiamycin sensitive mutant of Escherichia coli recipient containing the gene on a high copy number plasmid.

J Clin Immunol, 1988 Nov, 8(6), 513 - 20
Immunomodulation by indoleamines: serotonin and melatonin action on DNA and interferon-gamma synthesis by human peripheral blood mononuclear cells; Arzt ES et al.; Different concentrations of indoleamines, serotonin and melatonin, inhibited phytohemagglutinin stimulated DNA synthesis . Thus, 10(-3) to 10(-4) M of either indoleamine acted at the optimal phytohemagglutinin concentration, while 10(-3) to 10(-7) M acted at suboptimal phytohemagglutinin levels . The serotonin effect was reversed by the serotonergic S1-S2 receptor antagonist methysergide but not by the S2 antagonist ketanserin . This indicates that only the S1 receptor is involved in the inhibitory effect . Inhibition of lymphoproliferation by indoleamines was also exerted on pokeweed mitogen and protein A from Staphylococcus aureus stimulations . Serotonin and melatonin also inhibited phytohemagglutinin and protein A from Staphylococcus aureus induction of interferon-gamma synthesis . The initial uptake of Ca2+ was not affected by indoleamines, suggesting that it is not the mechanism of their inhibitory effects . As interferon-gamma induced tryptophan uptake by T lymphocyte- and macrophage-depleted populations, and tryptophan is the metabolic precursor of serotonin and melatonin, a new immunoregulatory circuit is postulated.

Eur J Immunol, 1988 Nov, 18(11), 1699 - 704
Effect of recombinant human interleukin 4 on spontaneous in vitro human IgE synthesis; Yang XD et al.; Recombinant human interleukin 4 (IL 4) alone enhanced the spontaneous IgE synthesis in cultures of peripheral blood leukocytes (PBL) from atopic patients as well as from nonatopic individuals, suggesting the existence of preactivated PBL sensitive for IL 4 . Preactivated cells were also obtained by stimulation with Staphylococcus aureus strain Cowan I (SAC) . However, co-stimulation of PBL by IL 4 with SAC or anti-IgM antibody and pokeweed mitogen did not result in an enhanced IgE synthesis . Optimal IL 4 concentrations for the induction of IgE synthesis coincided with optimal proliferative responses in PBL . The effect of IL 4 was not isotype specific, and in terms of protein even more IgG and IgM antibodies were formed . The effect of IL 4 on IgE synthesis was counteracted by very low concentrations of interferon-gamma (IFN-gamma), suggesting that both IL 4 and IFN-gamma might be decisive cytokines for the human in vitro IgE synthesis.

Blood, 1988 Nov, 72(5), 1553 - 9
Diversity in inhibitory effects of IFN-gamma and IFN-alpha A on the induced DNA synthesis of a hairy cell leukemia B lymphocyte clone reflects the nature of the activating ligand; Mongini P et al.; A hairy cell leukemia population was used as a clonal model for studying the direct immunomodulatory effects of recombinant interferon-alpha A (rIFN-alpha A) and rIFN-gamma on human B-cell proliferation . The leukemic cell population KON was notably quiescent when incubated in medium alone but was induced to significant in vitro DNA synthesis when cultured with any of four activators of human B cells: anti-IgM antibody, Staphylococcus aureus cells (SAC), phorbol myristate acetate (PMA), or B-cell growth factor (BCGF) . While both rIFN-gamma and rIFN-alpha A exhibited suppressive effects on these responses, their inhibitory patterns were distinct and reciprocal . Thus, rIFN-gamma exclusively suppressed anti-IgM-and SAC-induced leukemic DNA synthesis, and rIFN-alpha A significantly suppressed only PMA- and BCGF-induced DNA synthesis . The effects of the rIFN preparations were ablated in the presence of IFN type-specific monoclonal antibodies . Kinetic analyses and pulsing studies revealed that inhibition was most notable when cells were exposed concomitantly to IFN and the activating ligand . That the diverse effects of IFN-gamma and IFN-alpha A are manifested on a single B-cell clone was confirmed by Southern blot analysis of restriction enzyme-digested KON cell DNA with a JH-specific probe . These studies suggest that the therapeutic potential of the two types of IFN may be influenced by the nature of the extracellular ligands in the leukemic mileau that promote leukemic clonal expansion.

J Clin Microbiol, 1988 Nov, 26(11), 2391 - 4
Identification of Escherichia coli serotype O157 strains by using a monoclonal antibody; Perry MB et al.; The O157 antigenic determinant of Escherichia coli serotype O157:H7, an important bacterial pathogen, resides in the polysaccharide portion of its cellular lipopolysaccharide component which, from structural studies, was identified as a linear polymer of a repeating tetrasaccharide unit composed of D-glucose, L-fucose, 2-acetamido-2-deoxy-D-galactose, and 4-acetamido-4,6-dideoxy-D-mannose residues (1:1:1:1) . Hybrid cells producing monoclonal antibodies against the E . coli O157 antigen were obtained by fusion of myeloma cells with lymphocytes from BALB/c mice immunized with killed E . coli O157:117 cells . Clones were selected for binding specificity with purified O polysaccharide . One monoclonal antibody used in direct slide agglutinations or in coagglutination reactions with Staphylococcus aureus Cowan 1 cells sensitized with the affinity column-purified antibody accurately detected all strains of E . coli O157 tested . This selected monoclonal antibody did not agglutinate E . coli strains such as E . coli O7 and E . coli O116 or other bacteria which are known to give positive agglutinations with conventional polyclonal E . coli antisera . These results indicate that the monoclonal antibody is a superior specific-typing reagent.

Zentralbl Bakteriol Mikrobiol Hyg {A}, 1988 Nov, 270(1-2), 76 - 82
Complex typing of methicillin-resistant Staphylococcus aureus (MRSA); Witte W et al.; To discriminate between methicillin-resistant Staphylococcus aureus from 5 nosocomial outbreaks and from sporadic nosocomial infections, the efficacy of a complex typing scheme by phage typing, biochemical typing, resistance phenotype, plasmid profiles, plasmid patterns and attribution of resistance determinants to the chromosome was studied . In addition to the International Basic Set and experimental phages 88-93, 10 experimental phages from the Public Health Laboratory Service, Colindale, London, were used for phage-typing . The 10 experimental phages from PHLS in particular, in combination with plasmid profiles and plasmid patterns, were of special discriminative value.

Jpn J Antibiot, 1988 Nov, 41(11), 1758 - 73
{Studies on imipenem/cilastatin sodium in the perinatal period}; Cho N et al.; Pharmacokinetic, bacteriological and clinical studies of imipenem/cilastatin sodium (IPM/CS) were carried out in the perinatal period, and the results obtained are summarized as follows: Effects of IPM on bacterial growth curves of Staphylococcus aureus (sensitive and resistant strains) and Escherichia coli (sensitive strains) in amniotic fluid were determined . The bactericidal effects of IPM increased in amniotic fluid and remarkable increases against resistant strains were demonstrated . IPM and CS were detected promptly after intravenous drip infusion to pregnant women, and reached peak levels shortly after administration in maternal plasma . Placental penetration of IPM and CS to the fetus was favorable . After intravenous drip infusion of 500 mg/500 mg of IPM/CS, drug concentrations in the umbilical cord plasma and the amniotic fluid exceeded MICs against main pathogenic organisms . Clinically, IPM/CS was effective in the treatment of perinatal infections without any side effect . The above results demonstrated that IPM/CS is a clinically useful antibiotic for the prophylaxis and the treatment of perinatal infections.

Circ Shock, 1988 Nov, 26(3), 257 - 65
Staphylococcus aureus-induced shock: a pathophysiologic study; Hinshaw LB et al.; The purpose of the present study was to determine the effects of lethal intravenous infusions of Staphylococcus aureus (SA) in adult dogs . Animals were maintained under anesthesia for 6 hr and observed until death following the 1-hr infusions of SA organisms . Findings unique to SA administration compared to those with Escherichia coli were the absence of significant necrosis of the mucosal intestinal glands of the small and large intestines; widespread intravascular colonization of bacteria in lung, heart, kidney and adrenal tissues often associated with neutrophil sequestration, microabscess formation, and necrosis; relative constant blood pressure, fibrinogen, fibrin degradation products (FDP), serum glutamic pyruvic transaminase (SGPT), blood (serum) urea nitrogen (BUN), creatinine, pH, and PO2, all of which remained relatively unchanged for 6 hr . Rapid early increases were observed in temperature, respiration rate, lactate, and hematocrit, while PCO2, platelet and white blood cell concentrations decreased . Results suggest unique qualitative differences in responses to Staphylococcal-induced shock compared to those caused by gram-negative bacteria.

J Bacteriol, 1988 Nov, 170(11), 5337 - 43
Cloning and expression in Escherichia coli of genes encoding a multiprotein complex involved in secretion of proteins from Staphylococcus aureus; Adler LA et al.; The genes encoding the multiprotein membrane-bound ribosomal protein (MBRP) complex (mrp genes), associated with membrane-bound ribosomes in Staphylococcus aureus, were cloned in Escherichia coli . All four components (molecular sizes 71, 60, 46, and 41 kilodaltons) of the MBRP complex were expressed from an 8.5-kilobase DNA fragment as judged by Western blot (immunoblot) analysis . The order of the individual genes within the cloned DNA fragment was determined by deletion mutagenesis and subcloning of various restriction fragments . Three RNAs, transcribed from the same DNA strand, were identified within the MBRP-coding region: one large RNA of approximately 5.9 kilobases, presumably coding for all four MBRP components, and two minor RNAs, coding for MBRP-71 and MBRP-60 . The two minor RNAs seemed to be transcribed from promoters within the large transcription unit . Attempts to make insertional inactivations of the mrp genes with an internal 600-base-pair DNA fragment of the MBRP-coding region as a target were unsuccessful, presumably because such insertions are lethal.

J Clin Microbiol, 1988 Nov, 26(11), 2395 - 401
Immunoblots, antimicrobial resistance, and bacteriophage typing of oxacillin-resistant Staphylococcus aureus; Mulligan ME et al.; An immunoblotting system was developed for typing of oxacillin-resistant Staphylococcus aureus . Clinical isolates recovered during a 40-month period at a single institution were evaluated with this typing scheme . Results were compared with susceptibility patterns and with bacteriophage typing results for 100 clinical isolates and with plasmid fingerprints for 14 isolates . Immunoblotting was found to be a useful method with good reproducibility that distinguished seven major groups of patient isolates that were clinically and epidemiologically related . Susceptibility patterns showed specific correlations with other typing results but were inferior to immunoblotting and phage typing for differentiating major groups of organisms . Plasmid profiles failed to distinguish two major groups that were readily identified by immunoblots and phage typing . There was evidence of increasing antimicrobial resistance of endemic hospital strains . Immunoblotting correlated well with phage typing, offered an alternative method for typing isolates that could not be typed by phage typing, and was superior to susceptibility testing and plasmid profiles for distinguishing different groups of oxacillin-resistant S . aureus at our institution.

Zh Mikrobiol Epidemiol Immunobiol, 1988 Nov, (11), 16 - 8
{Isolation of strains of a new ecological variant of Staphylococcus aureus from asses}; Orzuev MI et al.; 47 coagulase-positive staphylococcal strains were isolated from the nasal smears of 175 healthy donkeys . In accordance with the schemes of Akatov--Devriese and Mayer-Witte--Akatov, 10.6% of the cultures were classified with the coagulase-positive species S . hyicus and 89.3%, with the species S . aureus . Out of S . aureus strains, 11.9% were found to have the characteristics of ecovar hominis, while 16.2% of the cultures could not be classified with definite ecovars . Most of the strains (71.4%) were found to differ from the known ecovars of S . aureus in their biological properties . For this reason, the above strains were classified with the new ecovar asinae . The authors propose to make the existing S . aureus identification scheme (the scheme of Mayer-Witte--Akatov) more complete by adding the tests for hyaluronidase and phosphatase.

Eur J Immunol, 1988 Nov, 18(11), 1753 - 60
Functional interaction between B cell subpopulations defined by CD23 expression; Armitage RJ et al.; Using the CD23 monoclonal antibody (mAb) MHM6 and sheep anti-mouse Ig bound to magnetic beads we have obtained highly purified populations of MHM6+ and MHM6- tonsil B cells . We have found that the increased expression of MHM6 reactivity seen on B cells after activation results from up-regulation of antigen on cells already weakly positive and not from expression of new antigen on the previously negative population . The strong proliferative responses of MHM6+ cells seen in the presence of anti-IgM (alpha mu) and interleukin 4 (IL4) or the CDw40 mAb G28-5, and with Staphylococcus aureus Cowan I (SAC), and to a lesser extent with the phorbol ester 12-O-tetradecanoylphorbol 13-acetate (TPA), resemble that seen among unfractionated B cells . In contrast, the MHM6- population cultured alone responds only weakly to alpha mu + G28-5 or SAC and exhibits virtually no response to alpha mu + IL4 or TPA . With all these mitogenic stimuli, tritiated thymidine uptake by the MHM6- population is augmented three- to sixfold by the addition of mitomycin C (MC)-treated MHM6+ cells . Pretreatment of cells with anti-leukocyte functional antigen 1 mAb has little effect on the subsequent proliferation of the MHM6- population but shows cell contact to be critical for the proliferation of MHM6+ cells . Such pretreatment has revealed that the functional interaction observed between MHM6+ and MHM6- cells is dependent on both cell contact and the presence of an MHM6+ cell-derived soluble component . We have found that addition of soluble CD23, purified from Epstein-Barr virus-transformed lymphoblastoid cell line supernatant, increases the proliferative response of MHM6- tonsil B cells to mitogenic stimuli in the presence of inactivated MHM6+ cells but has no effect on proliferation when MHM6+ cells are absent . By way of contrast to normal B lymphocytes, we have examined functional responses of prolymphocytic leukemia (PLL) B cells . Although these cells, when freshly isolated, show comparable levels of CD23 expression to normal B cells, this expression is not increased upon activation . In addition, in contrast to normal B cells, the PLL MHM6- population cultured alone shows a strong proliferative response to various mitogenic stimuli, comparable to that of MHM6+ or unfractionated cells, and this response is not augmented by the addition of MC-treated MHM6+ cells . Thus, a novel functional interaction is described between normal, but not leukemic, B cell populations defined by their expression of CD23.(ABSTRACT TRUNCATED AT 400 WORDS)

J Vasc Surg, 1988 Nov, 8(5), 600 - 5
Prevention of vascular prosthetic infection with an antibiotic-bonded Dacron graft; Shue WB et al.; Surfactant-mediated antibiotic bonding was used in an animal model of aortic prosthetic infection . Control grafts, control plus parenteral oxacillin, and oxacillin-bonded Dacron grafts were challenged by local inoculation with Staphylococcus aureus . Ninety percent of controls, 80% of parenteral antibiotic recipients, and only 30% of antibiotic-bonded Dacron grafts became infected (p less than 0.01, p less than 0.03) . Antibiotic-bonded grafts were also superior in terms of suture line cultures and patency . In separate experiments in a subcutaneous pouch model, antibiotic bonding significantly improved the median infective dose of Dacron over that of controls and Dacron soaked in cephalosporin . These studies demonstrate that antibiotic-bonded Dacron implants are highly resistant to infection . A multicenter clinical trial is planned.

Arch Dermatol, 1988 Nov, 124(11), 1691 - 700
Topical antibiotics in dermatology; Hirschmann JV; Topical antibiotics are safe and effective in certain conditions, primarily acne, rosacea, and nasal carriage of Staphylococcus aureus . They are useful in impetigo only when it is of limited extent . Their efficacy in other pyodermas is unclear, although mupirocin is probably effective in many cases . In "infected eczema" that does not require systemic therapy they seem to add little to what topical corticosteroids alone achieve . They are ineffective in reducing the incidence of significant infection with indwelling intravenous catheters . They are safe preparations, but extensive use, especially in closed populations, may encourage the emergence of resistant bacteria.

Proc Natl Acad Sci U S A, 1988 Nov, 85(21), 8012 - 6
Ethylene-regulated expression of a tomato fruit ripening gene encoding a proteinase inhibitor I with a glutamic residue at the reactive site; Margossian LJ et al.; We report the isolation from tomato (Lycopersicon esculentum) of an ethylene-responsive member of the proteinase inhibitor gene family . DNA sequence analysis of a full-length cDNA clone indicates that the ethylene-responsive gene is distantly related to the tomato proteinase inhibitor I gene, having 53% sequence identity . The predicted amino acid sequence reveals 47% and 45% sequence identity with the tomato and potato proteinase inhibitor I polypeptides, respectively . Additionally, the ethylene-responsive inhibitor has evolved a completely different pattern of gene expression and inhibitory specificity than other members of the inhibitor I family . Gel blot hybridization experiments show that, unlike the tomato proteinase inhibitor I gene, it is not induced in wounded leaves . In contrast, it is activated by the plant hormone ethylene in leaves and during fruit ripening . Furthermore, the ethylene-responsive inhibitor exhibits a novel reactive site, having glutamic acid as the P1 residue . This suggests that the ethylene-responsive proteinase inhibitor does not react with chymotrypsin, as does proteinase inhibitor I, but that it reacts with proteolytic enzymes that cleave at glutamic residues, such as the Staphylococcus aureus V8 proteinase, for which no inhibitors are known . Finally, isolation and analysis of a genomic clone reveals that the ethylene-responsive proteinase inhibitor gene is tightly linked to another, yet unidentified, coordinately expressed gene . We discuss these results with regard to the function and evolution of proteinase inhibitor genes in tomato.

Res Rep Health Eff Inst, 1988 Nov, (20), 1 - 38
Modulation of pulmonary defense mechanisms against viral and bacterial infections by acute exposures to nitrogen dioxide; Jakab GJ; The scientific literature suggests that ambient levels of nitrogen dioxide increase susceptibility to respiratory infections . However, this association has not been conclusively demonstrated . The epidemiologic data regarding this relationship are inconclusive because these studies have used parameters of "acute respiratory illness" that are not necessarily related to infectious episodes . Previous animal studies have used either mortality after bacterial infection with virulent bacteria or decreased rate of intrapulmonary killing of bacteria with low virulence . Studies using appropriate bacterial and viral challenge organisms, with morbidity as an endpoint, provide a better basis for extrapolation to humans . The investigations in animals suggest a relationship between nitrogen dioxide and increased susceptibility to respiratory infection, but studies in which functional parameters of host resistance to such infections have been used are few . The aim of this work was to determine the threshold level of acute nitrogen dioxide exposure that would induce increased susceptibility to, and increased severity of, viral and bacterial infections . Physiologic parameters of host resistance to respiratory infections were used as endpoints . A composite picture was developed of dose-response relationships between nitrogen dioxide and the impairment of a spectrum of defense parameters in the murine respiratory tract against viral and bacterial challenges . The salient findings of this study are as follows: (1) the intrapulmonary killing of Staphylococcus aureus was impaired at 5 ppm of nitrogen dioxide; (2) this effect was found at 2.5 ppm or less when nitrogen dioxide exposure was superimposed on lungs predisposed to lowered resistance through immunosuppression with corticosteroids; (3) the adverse effect of nitrogen dioxide occurred at lower concentrations when exposure followed bacterial challenge; and (4) during the course of murine Sendai virus infection, exposure to nitrogen dioxide for four hours per day did not alter the infection in the lungs, but rather it enhanced lung pathology . The implications of these findings are that the antibacterial defenses of the lungs are susceptible to the inhibiting effects of short acute exposures of lower concentrations of nitrogen dioxide when the lungs are predisposed by bacteria present or, even more so, by immunosuppression . The alveolar macrophage phagocytic system is the defense component of the lungs that is most susceptible to the adverse effects of nitrogen dioxide . The finding that nitrogen dioxide increases virus-associated lung damage suggests that the increased severity of the disease process results from the proliferation of the virus to high titers, rather than from alterations of the infective process.

J Gen Microbiol, 1988 Nov, 134 ( Pt 11), 2857 - 66
Structural and evolutionary relationships of beta-lactamase transposons from Staphylococcus aureus; Gillespie MT et al.; A comparison of the beta-lactamase elements detected on three classes of large plasmids together with the chromosomes of penicillin-resistant Staphylococcus aureus revealed substantial physical and genetic relatedness . In most cases, beta-lactamase production could be associated with the presence of a DNA segment of approximately 6.7 kb . Analysis showed that the plasmid-borne determinants constitute nearly identical transposons or transposon-like elements . An element indistinguishable from one of these, Tn4002, which is carried by the pSK1 family of plasmids in clinical isolates from Australian hospitals, was also identified on the staphylococcal chromosome and is implicated in an evolutionary cycle of transposition between chromosomal and extrachromosomal sites in Australian strains of multiresistant S . aureus.

Biochem J, 1988 Nov 1, 255(3), 817 - 24
Characterization of the autophosphorylation of chicken gizzard caldesmon; Scott-Woo GC et al.; Caldesmon, an actin- and calmodulin-binding protein of smooth muscle, is a protein serine/threonine kinase capable of Ca2+/calmodulin-dependent autophosphorylation {Scott-Woo & Walsh (1988) Biochem . J . 252, 463-472} . Phosphorylation nullifies the inhibitory effect of caldesmon on the actin-activated Mg2+-ATPase activity of smooth-muscle myosin {Ngai & Walsh (1987) Biochem . J . 244, 417-425} . We have characterized the kinase activity of caldesmon of chicken gizzard smooth muscle . Autophosphorylation requires Ca2+/calmodulin, but is unaffected by other second messengers (Ca2+/phospholipid/diacylglycerol, cyclic AMP or cyclic GMP), and is inhibited by the calmodulin antagonists chlorpromazine and compound 48/80, with 50% inhibition at 39.8 microM and 12.0 ng/ml respectively . Half-maximal activation of autophosphorylation occurs at 60-80 nM-Ca2+ and 0.14 microM-calmodulin, and maximal activity at 0.14-0.18 microM-Ca2+ and 1 microM-calmodulin; activation is gradually lost at higher Ca2+ and calmodulin concentrations . Autophosphorylation is pH-dependent, with maximal activity over the range pH 7-9, and requires free Mg2+ in addition to the MgATP2- substrate . The Km for ATP is 15.6 +/- 4.1 microM (mean +/- S.D., n = 4), and kinase activity is inhibited by increasing ionic strength {half-maximal inhibition at I = 0.094 +/- 0.009 M (mean +/- S.D., n = 4)} . Autophosphorylation does not affect the rate of hydrolysis of caldesmon (free or bound to calmodulin) by alpha-chymotrypsin . However, a slight difference in peptides generated from phospho- and dephospho-forms of caldesmon is observed . The binding of phospho- or dephospho-caldesmon to F-actin protects the protein against chymotryptic digestion, but does not alter the pattern of peptide generation . Characterization of proteolytic fragments of caldesmon generated by alpha-chymotrypsin and Staphylococcus aureus V8 protease enables localization of the phosphorylation sites and the kinase active site within the caldesmon molecule.

Mol Microbiol, 1988 Nov, 2(6), 709 - 17
Nucleotide sequence analysis of 2''-aminoglycoside nucleotidyl-transferase ANT(2'') from Tn4000: its relationship with AAD(3'') and impact on Tn21 evolution; Schmidt FR et al.; Aminoglycoside 2''-O-nucleotidyltransferase (AAD(2'')) mediates bacterial resistance to dibekacin, gentamicin, kanamycin, sisomicin and tobramycin . Its coding sequence, aadB, is part of Tn21-related transposon, Tn4000 . Nucleotide sequence analysis revealed the presence of an open reading frame capable of specifying a protein of 177 amino acids with a calculated molecular weight of 21,240 . The predicted amino acid sequence revealed up to 27% homology to that of three nucleotidyltransferases of type AAD(3''), which are widely distributed among Gram-negatives, and to the AAD(9) from Staphylococcus aureus transposon Tn554 . The regions flanking aadB suggest that its insertion into Tn21 arose from a site-specific recombination event adjacent to the aadA gene.

J Virol, 1988 Nov, 62(11), 4303 - 6
Receptors for Theiler's murine encephalomyelitis virus: characterization by using rabbit antiviral antiserum; Rubio N et al.; An immunological assay was developed to characterize the binding of Theiler's murine encephalomyelitis virus to BHK-21 cell receptors . After absorption of the virus and formaldehyde fixation, rabbit antibodies and Staphylococcus aureus protein A labeled with 125I formed a specific complex on the surfaces of the cells . The optimal multiplicity of infection in this system was 10 PFU per cell . The virus was internalized at 33 and 37 degrees C, but internalization did not take place at 25 or 4 degrees C . The binding was proportional to the number of cells and was significant within 30 s . Cell surface receptors were still active after fixation, and only intact viruses were bound, as demonstrated by the lack of binding of the purified, isolated virion proteins VP1, VP2, and VP3.

Zh Mikrobiol Epidemiol Immunobiol, 1988 Nov, (11), 85 - 9
{The specificity of erythrocyte staphylococcal diagnostic agents in relation to the presence of protein A in sensitin preparations}; Deriabin PN et al.; Protein A has been detected in Staphylococcus aureus muriatic extracts by means of the passive hemagglutination test and the passive hemagglutination inhibition test with IgG erythrocyte diagnosticum . The absence of perceptible amounts of protein A, capable of interaction with the Fc fragment of IgG, on the surface of erythrocyte diagnosticum obtained from S . aureus muriatic extract with the use of amidol has been proved by different methods . The use of antierythrocytic IgG antibody bridge has shown that protein A in S . aureus muriatic extract is bound with other species-specific antigens, in particular with teichoic acid.

Res Vet Sci, 1988 Nov, 45(3), 353 - 9
Modulation of IgG subclass expression during antibody responses in sheep; Kerlin RL et al.; Experiments were undertaken to investigate IgG subclass expression during epitope-specific antibody responses in sheep . Animals were immunised with conjugates of bovine serum albumin-DNP (BSA-DNP), or killed Staphylococcus aureus-DNP (Sa-DNP) alone or with muramyl dipeptide (MDP), dextran sulphate (DXS), or staphylococcal exotoxins (toxin) . Sheep received two injections of the same preparation intracutaneously at six weeks interval . Total and IgG subclass-specific anti-DNP, and anti-carrier (BSA or S aureus) antibody levels were measured by ELISA in blood taken at weekly intervals before and after immunisation . Anti-DNP antibody levels in animals given Sa-DNP alone were considerably greater than in those immunised with BSA-DNP alone . Toxin suppressed antibody responses to DNP and both carriers; MDP suppressed anti-hapten antibody responses below the levels obtained with antigen alone . Neither toxin nor MDP significantly altered the IgG subclass profile of antibody to DNP bound to either carrier . DXS did not significantly change total levels of anti-DNP antibody measured in sheep given BSA-DNP or S aureus-DNP . However, DXS promoted IgG1 and suppressed IgG2 anti-DNP antibody responses during the secondary response to Sa-DNP but not to BSA-DNP.

Infect Immun, 1988 Nov, 56(11), 2861 - 5
In vivo activation of neutrophil function in hamsters by recombinant human granulocyte colony-stimulating factor; Cohen AM et al.; The in vivo effect of Escherichia coli-derived recombinant human granulocyte colony-stimulating factor on neutrophil function was studied in golden Syrian hamsters . Significant increases in superoxide generation and specific binding of N-formylmethionyl-leucyl-phenylalanine were observed in neutrophils isolated 4 h following a single subcutaneous injection of the factor (30 micrograms/kg) . However, phagocytotic activity was not significantly stimulated in hamsters treated with the factor . Recombinant human granulocyte colony-stimulating factor hastened the recovery of peripheral neutrophil counts in animals made leukopenic by prior treatment with cyclophosphamide . Beginning several hours after infection, resistance to lethal infection following intraperitoneal injection of Staphylococcus aureus was increased when neutropenic animals were treated daily with the factor . This protective effect was associated with increased peritoneal neutrophil counts and a decreased incidence of positive peritoneal bacterial cultures at 24 h after the start of treatment . These results suggest that recombinant human granulocyte colony-stimulating factor may be a useful adjunct in the treatment of bacterial infections in neutropenic patients.

Ann Intern Med, 1988 Oct 15, 109(8), 619 - 24
Right-sided Staphylococcus aureus endocarditis in intravenous drug abusers: two-week combination therapy; Chambers HF et al.; STUDY OBJECTIVE: To determine the efficacy of short-course combination regimens for selected cases of Staphylococcus aureus endocarditis in intravenous drug abusers . DESIGN: Open study of nafcillin and tobramycin or vancomycin and tobramycin administered for 2 weeks with no further therapy . SETTING: County hospital . PATIENTS: Consecutive sample of 53 intravenous drug abusers with relatively uncomplicated right-sided S . aureus endocarditis, defined by clinical and echocardiographic criteria, and without renal insufficiency, extrapulmonary metastatic infectious complications requiring prolonged therapy or surgery for cure, meningitis, methicillin-resistant organism, aortic or mitral valve involvement, or pregnancy . INTERVENTIONS: Nafcillin, 1.5 g intravenously every 4 hours, plus tobramycin, 1 mg/kg body weight intravenously every 8 hours, administered for 2 weeks . Vancomycin, 30 mg/kg per day intravenously, in two or three divided doses, was used instead of nafcillin for patients allergic to penicillin . MEASUREMENTS AND MAIN RESULTS: Forty-seven of 50 patients (94%; 95% CI, 87 to 99+) treated with the nafcillin and tobramycin combination were cured . Only 1 of 3 patients treated with vancomycin plus tobramycin (33%, 95% CI, 2 to 86) was cured . CONCLUSIONS: Selected patients with S . aureus endocarditis can be treated safely and effectively with a 2-week course of nafcillin plus tobramycin . Only one of three patients treated with vancomycin plus tobramycin was cured, but three patients are too few to define with confidence the efficacy of this regimen.

Eur J Biochem, 1988 Oct 15, 177(1), 125 - 33
Refolding of bacteriorhodopsin . Protease V8 fragmentation and chromophore reconstitution from proteolytic V8 fragments; Sigrist H et al.; Staphylococcus aureus protease V8 cleaves bacteriorhodopsin to two main fragments, V-1 and V-2 . Proteolytic digestion of the purple membrane integrated protein is carried out in the presence of limited amounts of sodium dodecyl sulfate (0.5 g detergent/g bacteriorhodopsin) . The fragment V-1 includes the arylisothiocyanate binding site (Lys41) . The V-2 fragment comprises the two C-terminal transmembrane segments of bacteriorhodopsin . Improved renaturation of bacteriorhodopsin and the ternary complex, reformed from its V8 proteolytic fragments, is attained by peptide extraction in chloroform/methanol/0.1 M ammonium acetate and subsequent incorporation into phospholipid/detergent micelles . In the presence of retinal, V8 fragments reform chromophoric ternary complexes . Light-adapted reconstituted chromophores absorb incident light at 560 nm . Protein secondary structures are partially conserved in the course of solvent extraction and are restored in the reconstituted system . Vesicles prepared from the reconstituted complexes show light-dependent proton translocation activity.

Biochim Biophys Acta, 1988 Oct 14, 962(3), 308 - 16
Mode of action of tetrahydrolipstatin: a derivative of the naturally occurring lipase inhibitor lipstatin; Borgstrom B; Tetrahydrolipstatin is a specific lipase inhibitor derived from lipstatin, a lipid produced by Streptomyces toxytricini . In addition to pancreatic lipase, it is shown in the present study that tetrahydrolipstatin also inhibits human gastric lipase, carboxyl ester lipase (cholesterol esterase) of pancreatic origin and the closely related bile-salt-stimulated lipase of human milk . It does not inhibit the exocellular lipase from Rhizopus arrhizus or a lipase recently isolated from Staphylococcus aureus . In the presence of a water-insoluble substrate, such as tributyrin, the inhibition has the characteristics of an irreversible inactivation of the uncompetitive type, thus indicating that an enzyme.substrate.inhibitor complex is formed, which cannot undergo further reaction to yield the normal product . This reaction probably takes place at the aqueous/oil interface of the substrate . In aqueous solution, in the absence of substrate, the inhibition of carboxyl ester lipase by tetrahydrolipstatin has the characteristics of being reversible, and finally becomes of a temporary nature analogues to the trypsin-trypsin inhibitor system . It is suggested that an enzyme-inhibitor complex of an acyl-enzyme type is formed that is slowly hydrolysed, with water as the final acceptor, leaving an intact enzyme and an inactive form of the inhibitor . The enzyme thus consumes the inhibitor, which undergoes a chemical conversion, as indicated by a change in mobility in an appropriate thin-layer chromatographic system, indicating an increase in hydrophilicity . Evidence is presented that the reaction product is an acid and that the functional group of tetrahydrolipstatin is the beta-lactone reacting with the active site of the enzyme.

Biochim Biophys Acta, 1988 Oct 12, 956(3), 224 - 31
Histone H1 structure probed by Staphylococcus aureus V8-proteinase; Bohm L et al.; Proteolytic digestion of calf thymus histone H1 with Staphylococcus aureus V8-proteinase under structuring conditions generates one major limit peptide P1 which consists of approx . 170 residues . Edman degradation establishes the N-terminal sequence as: Leu-Ile-Thr-Lys-Ala-Val-Ala-Ala-Ser-Lys . Chymotryptic fingerprinting shows that the C-terminal part of the H1 molecule is fully preserved . The peptide therefore comprises the residues H1 (42-210) . The Glu-41 cleavage is extremely unusual as it occurs in the structured G-domain which is known to be resistant to proteinases (Hartman, P . G., Chapman, G . E., Moss, T . and Bradbury, E . M . (1977) Eur . J . Biochem . 77, 45-71; Bohm, L., Sautiere, P., Cary, P . D . and Crane-Robinson, C . (1982) Biochem . J . 203, 577-582) . The V8-proteinase cleavage product H1 (42-210) shows only 20% folding as compared to 95-99% folding shown by the peptides H1 (34-121), H1 (31-210) and H1 (33-210) . Folding of the G-domain thus critically depends upon the presence of the eight residues 33-41 amongst which the Gly-Pro-Pro sequence at position 36-38 and a beta-turn predicted at position 35 are considered to be particularly important . The location of the cleavage site in the G-domain renders Staphylococcus aureus V8-proteinase suitable as a structural probe.

FEBS Lett, 1988 Oct 10, 238(2), 419 - 23
Insulin-induced decrease in 5'-nucleotidase activity in skeletal muscle membranes; Klip A et al.; Insulin releases inositol phosphoglycans from myocytes in culture {(1986) Science 233, 967-972}, which display insulinomimetic activity . Because 5'-nucleotidase is anchored to the membrane through inositol-containing phospholipid glycans, we investigated whether insulin could release the enzyme from the membrane . Membranes prepared from hindquarter muscles of rats perfused with insulin showed a 23% decrease in 5'-nucleotidase activity . Isolated membranes from muscle exposed to insulin in vitro also showed a small but reproducible decrease (9%) in 5'-nucleotidase activity relative to unexposed controls . Phospholipase C from Staphylococcus aureus released 60% of the membrane-bound 5'-nucleotidase . We propose that insulin may activate an endogenous phospholipase C that cleaves phospholipid-glycan-anchored proteins.

Biochim Biophys Acta, 1988 Oct 7, 971(3), 266 - 74
The bactericidal effects of the respiratory burst and the myeloperoxidase system isolated in neutrophil cytoplasts; Odell EW et al.; Neutrophil polymorphonuclear leucocytes kill bacteria by oxygen-dependent and oxygen-independent mechanisms . Many potentially toxic mechanisms have been described, but the complexity of the phagosomal environment and the synergy between oxidative and non-oxidative systems hamper the investigation of individual bactericidal mechanism in whole cells . Neutrophil cytoplasts are greatly depleted of granule proteins and permit the investigation of the bactericidal effects of the respiratory burst in isolation . In this study they have been used to examine the role of the respiratory burst and myeloperoxidase in oxygen-dependent killing of Staphylococcus aureus . Cytoplasts generated oxygen radicals at comparable rates to human neutrophils and phagocytosed but did not kill S . aureus . The selective reconstitution of the myeloperoxidase-hydrogen peroxide-halide system by coating bacteria with myeloperoxidase conferred on cytoplasts the ability to kill intracellular bacteria . However, extracellular killing by diffusible bactericidal factors was not detected in this system.

J Biol Chem, 1988 Oct 5, 263(28), 14552 - 8
Biosynthesis of the plasma membrane H+-ATPase of Neurospora crassa; Aaronson LR et al.; The plasma membrane H+-ATPase of Neurospora is a 100-kDa integral membrane protein which appears, on the basis of hydropathy analysis of its amino acid sequence, to span the lipid bilayer at least eight times . To investigate the assembly and processing of the ATPase, a full-length cDNA has been constructed for use in in vitro transcription and translation experiments . Comparison of three different forms of the ATPase (nascent protein, nascent protein cotranslationally inserted into membranes, and mature protein) revealed no difference in electrophoretic mobility . Furthermore, the nascent and mature forms gave identical peptide patterns after partial proteolysis with Staphylococcus aureus V8 protease, suggesting that the ATPase does not contain an NH2-terminal signal peptide which is cleaved upon membrane insertion . Consistent with this interpretation, the NH2-terminal peptide has been purified from a tryptic digest of the ATPase and found to lack only the initiator methionine residue; the penultimate alanine is acetylated based on analysis by fast atom bombardment mass spectroscopy . Although the ATPase contains one potential site of N-linked glycosylation, its electrophoretic mobility was unchanged following digestion with endoglycosidase H and it did not incorporate {3H}mannose or bind concanavalin A . Thus, the Neurospora plasma membrane-ATPase appears to undergo minimal post-translational processing, and its membrane insertion is probably mediated by internal sequences.

Biochemistry, 1988 Oct 4, 27(20), 7773 - 7
Complete amino acid sequence of the thioesterase domain of chicken liver fatty acid synthase; Yang CY et al.; The complete amino acid sequence of thioesterase domain of chicken liver fatty acid synthase has been determined by sequencing peptides produced by trypsin, Staphylococcus aureus V8 protease, and cyanogen bromide cleavage . The thioesterase domain consists of 300 amino acid residues . All of the tryptic peptides of the thioesterase domain were isolated and sequenced, except the segment covered from position 109 to position 124 . Peptides resulting from digestion by Staphylococcus aureus V8 protease and cyanogen bromide cleavage filled the missing part and overlapped the complete sequence of the entire thioesterase domain . The NH2 terminus of the thioesterase domain was determined to be lysine by sequencing the whole domain up to 20 residues while the COOH terminus was identified as serine through carboxyl peptidase Y cleavage . The active site of the thioesterase domain of chicken fatty acid synthase was suggested to be the serine on position 101 according to its homology with other serine-type esterases and proteases which have a common structure of -Gly-X-Ser-Y-Gly- with the variable amino acids X and Y disrupting the homology.

J Immunol Methods, 1988 Oct 4, 113(1), 101 - 11
Cell-specific heterogeneity in sensitivity of phosphatidylinositol-anchored membrane antigens to release by phospholipase C; Low MG et al.; Cell surface antigens thought to be linked to the membrane via phosphatidylinositol (PI) are incompletely, and variably, released by treatment with phosphatidylinositol-specific phospholipase C (PI-PLC) . The basis for this was investigated with cloned tumor cell lines and PI-PLCs isolated from two species of bacteria . Residual Thy-1 antigen, which was detectable by flow cytometry, remained on all thymoma cell lines after exposure to very high concentrations of either purified enzyme . A majority of the presumptive PI anchored molecules on all of the cell lines was sensitive to release by PI-PLC derived from Bacillus thuringiensis . However, cell lines differed dramatically in the ease with which PI-PLC from Staphylococcus aureus liberated the same surface antigens . This heterogeneity was determined at the single cell level because at least five different PI-anchored antigens exhibited similar behavior on a given cell line or transfected subclones of it . The two phospholipases differed with respect to molecular mass, serological cross-reactivity and sensitivity to inhibition by NaCl and detergents . These observations suggest that the two types of PI-PLC may have distinct substrate specificities or sensitivities to environmental conditions which account for the difference in their ability to act on PI-anchored proteins in particular cell types . Such enzymes should continue to be important tools for investigating the method and significance of attachment of lymphocyte surface glycoproteins . In particular, the S . aureus PI-PLC can be used to demonstrate and investigate a previously unrecognized heterogeneity in cells which express PI-anchored molecules.

Obstet Gynecol, 1988 Oct, 72(4), 559 - 64
Development of wound infections among women undergoing cesarean section; Emmons SL et al.; Sixty consecutive wound infections were studied among 1104 women undergoing cesarean section . Wound infections caused by cervical-vaginal flora were associated with prolonged labor, particularly with greater duration of fetal monitoring and number of vaginal examinations, and with organisms isolated from the endometrium at cesarean section . In contrast, women with wound infections caused by Staphylococcus aureus had neither prolonged labor nor S aureus isolated at cesarean section . The 25% of wound infections associated with S aureus represent potentially preventable conditions that presumably arise from exogenous sources.

Immunol Lett, 1988 Oct, 19(2), 153 - 7
Up-regulation of receptors for IgA on activated human B lymphocytes; Millet I et al.; Expression of receptors for the Fc part of IgA (Fc alpha R) by T lymphocytes was recently shown to be up-regulated after activation by T cell mitogens in the absence of IgA . We describe a similar increase on activated human B lymphocytes . Fc alpha R were determined by labelling with human secretory IgA (0.5 mg/ml) and flow cytometry analysis after staining with fluoresceinated goat anti-IgA or goat anti-secretory component F(ab')2 fragments . B-enriched cell suspensions were prepared from peripheral blood or tonsils and activated by Staphylococcus aureus Cowan I, anti-IgM antibodies or E . coli lipopolysaccharide . All three activators increased the percentage of Fc alpha R positive cells although only the former induced significant DNA synthesis . Finally recombinant interleukin 1 (10 nM) and interleukin 2 (10 IU/ml) but not interleukin 4 (300 units/ml) nor low-molecular-weight B cell growth factor induced an increase of Fc alpha R expression . The data show that Fc alpha R can be up-regulated on human B cells in the absence of exposure to IgA.

Aust Paediatr J, 1988 Oct, 24(5), 275 - 9
The changing pattern of severe neonatal staphylococcal infection: a 10-year study; Tam AY et al.; Forty-two cases of severe staphylococcal infection occurring over a 10-year period in the neonatal unit at Queen Mary Hospital are described . There was a 4.5-fold increase in incidence in the latter half of the study period, when methicillin-resistant Staphylococcus aureus (MRSA) emerged . The isolated MRSA were also resistant to gentamicin, but sensitive to vancomycin, fusidic acid, co-trimoxazole and amikacin . Comparison between MRSA and methicillin-sensitive cases showed that the former was associated with a longer hospital stay after diagnosis . Overall mortality was 9.5% . Two cases with meningitis died . MRSA is at least as virulent as its methicillin-sensitive counterparts . The treatment implications of severe neonatal staphylococcal infection are discussed.

J Appl Bacteriol, 1988 Oct, 65(4), 329 - 37
Sensitivity of methicillin-resistant Staphylococcus aureus strains to some antibiotics, antiseptics and disinfectants; Al-Masaudi SB et al.; The effects of antibiotics, antiseptics and disinfectants against some methicillin-resistant (MRSA) and methicillin-sensitive (MSSA) Staphylococcus aureus strains have been studied . The MRSA and MSSA strains were equally sensitive to phenols, esters of para(4)-hydroxybenzoic acid and chlorhexidine but MRSA strains were slightly more resistant to quaternary ammonium compounds and considerably more so to dibromopropamidine isothionate . Some MRSA strains were also resistant to phenylmercuric nitrate (but not another organomercurial, thiomersal), mercuric chloride and cadmium chloride . All MRSA strains produced beta-lactamase . Strains from the Royal Free Hospital, London were highly resistant to beta-lactam antibiotics, erythromycin, trimethoprim and tetracyclines but were sensitive to other antibiotics . One strain from the University Hospital of Wales, Cardiff was resistant to gentamicin but sensitive to tetracycline and trimethoprim.

J Rheumatol, 1988 Oct, 15(10), 1539 - 46
In vitro immunoglobulin production by lymphocytes of patients with juvenile rheumatoid arthritis: effects of Staphylococcus aureus Cowan I stimulation and monocyte depletion; Oen K et al.; Pokeweed mitogen (PWM) and Staphylococcus aureus Cowan I (SAC) were used to stimulate in vitro IgG and IgM production by lymphocytes of 27 patients with juvenile rheumatoid arthritis (JRA) . Twelve had reduced stimulation indices for PWM stimulated cultures of T and non-T cells . Stimulation with SAC resulted in increased IgM production in half (5/10); and partial removal of monocytes resulted in improved PWM induced IgM production in 5/7 . IgG production was less easily improved . The results of our study suggest that while PWM induced Ig production may be reduced, B cells responding to SAC may function normally in some patients with JRA . In others, monocyte mediated suppression may account for reduced responses to PWM.

Am J Infect Control, 1988 Oct, 16(5), 185 - 92
Cost of nosocomial infection: relative contributions of laboratory, antibiotic, and per diem costs in serious Staphylococcus aureus infections; Wakefield DS et al.; This study reports an analysis of the relative importance of laboratory antibiotic, and per diem costs of caring for 58 patients with serious Staphylococcus aureus nosocomial infections . Laboratory costs accounted for 2%, antibiotics for 21%, and per diem costs for 77% of total infection-related costs . Only 45% of patients were hospitalized for additional days specifically because of infection, but these patients stayed an average of 18 extra days . Nosocomial infections with S . aureus resistant to penicillinase-resistant penicillins (PRP) were more frequently associated with additional infection-related days of hospitalization than were PRP-susceptible infections . The cost of PRP-resistant infections was also significantly greater than PRP-susceptible infections, primarily because of the costs of additional days of hospitalization . Rational strategies to control costs of nosocomial infection should focus on two approaches: (1) prevention and (2) reduction of acute hospital days attributable to infections.

Eur J Immunol, 1988 Oct, 18(10), 1491 - 8
Antigen-independent activation of T cells mediated by a novel cell surface heterodimer (Tp135-145); Isler P et al.; A new heterodimeric structure, Tp135-145, which can mediate interleukin 2 (IL2) production and Ca2+ mobilization by Jurkat cells is described . This structure was identified by a monoclonal antibody, MX24, on the surface of either T3/TcR+ or T3/TcR- human T cell lines as well as on B cell lines . Biochemical studies showed that antibody MX24 precipitated two polypeptide chains of 135 and 145 kDa, respectively, in lysates from 125I-labeled T cells . After reduction the 135-kDa polypeptide chain shifted to 140 kDa, whereas the molecular mass of the other polypeptide remained unchanged . The apparent molecular masses of the desialylated polypeptides differed by 5 kDa . No common peptide fragments between the two polypeptide chains were found after limited proteolysis by Staphylococcus aureus V8 protease . The expression of Tp135-145 was independent of the expression of the T3/TcR molecular complex . Incubation of Jurkat cells with anti-TcR or anti-T3 monoclonal antibody induced complete modulation only of the T3/TcR complex but not of Tp135-145 . Conversely complete modulation of Tp135-145 was observed after incubation of these cells with MX24 antibody . Functional studies showed that anti-Tp135-145 antibody MX24 induced high levels of IL2 production in Jurkat cells . In addition, incubation of these cells with MX24 resulted in Ca2+ mobilization from internal stores . In peripheral blood, Tp135-145 was found to be expressed by 39%-76% of resting T cells in individual donors . Two-color flow microfluorimetry showed that the Tp135-145+ cells were equally distributed on the CD4+ and CD8+ subsets . Incubation of peripheral blood T cells with antibody MX24 resulted in IL2 production and cell proliferation . Taken together these results suggest that Tp135-145 is a novel surface molecule involved in antigen-independent pathway of T cell activation.

J Gen Virol, 1988 Oct, 69 ( Pt 10), 2505 - 16
Humoral and cytotoxic T cell immune responses to internal and external structural proteins of simian virus 5 induced by immunization with solid matrix-antibody-antigen complexes; Randall RE et al.; Monoclonal antibodies (MAbs) were complexed to solid matrices and used to purify virus proteins from simian virus 5 (SV5)-infected tissue culture cells, ideally in such a way as to bind equimolar amounts of antigen to antibody . The resulting solid matrix-antibody-antigen (SMAA) complexes were then used as immunogens and successfully induced specific humoral and cytotoxic T cell responses . By attaching more than one MAb to the solid matrix and using such complexes to purify the respective proteins multivalent immunization was achieved . Analysis of the cytotoxic T cell response of immunized animals indicated that both surface and internal SV5 structural proteins can act as target antigens . Immunization with SMAA complexes, in the absence of adjuvant, induced higher levels of antibody than the antigen alone precipitated on alum . However, immunization with SMAA complexes resulted in relatively less antibody being produced to the antigenic determinant through which the protein is coupled, via antibody, to the SMAA complex compared with the amount of antibody produced against other antigenic determinants on that protein . The particulate solid matrix used to form the SMAA complexes in most of the experiments was a 'fixed' and killed suspension of the Cowan A strain of Staphylococcus aureus, although preliminary results indicated that Protein A-Sepharose could also be used as a solid matrix . Prior immunization with S . aureus alone did not reduce the level of the immune response to the appropriate antigen on subsequent immunization with S . aureus-antibody-antigen complexes . In fact on immunization of mice with these complexes the level of antibody produced to the S . aureus matrix itself was less when S . aureus-antibody-antigen complexes were used as immunogens than when S . aureus or S . aureus-antibody complexes were used . Furthermore, rabbits immunized with S . aureus-mouse MAb-antigen complexes showed a vigorous immune response to the antigen.

J Exp Med, 1988 Oct 1, 168(4), 1321 - 37
Interleukin 4 inhibits the proliferation but not the differentiation of activated human B cells in response to interleukin 2; Defrance T et al.; The combined effect of IL-4 and IL-2 on proliferation of anti-IgM antibody or Staphylococcus aureus strain Cowan I (SAC)-preactivated B cells was investigated . It was observed that in most cases, rIL-2 used at optimal concentration induced higher levels of tritiated thymidine ({3H}TdR) uptake than rIL-4 used at optimal concentration . When rIL-4 and rIL-2 were added together, it was repeatedly found that B cell proliferation induced by rIL-2 was significantly reduced and was, in most cases, comparable with the proliferation induced by rIL-4 alone . Cell cycle studies demonstrated that rIL-4 significantly reduced the number of cells entering S and G2/M phases of the cell cycle upon rIL-2 stimulation . B cell blasts preincubated for 24 or 48 h with rIL-4 displayed a reduced proliferation in response to rIL-2 . In contrast, preculture of resting B cells with rIL-4 did not impair their subsequent proliferation in response to rIL-2 plus insolubilized anti-IgM antibody . This suggests that rIL-4 can only exert its inhibitory effect once B cells have received an activation signal . The differentiative activity of rIL-2 measured on B cell blasts preactivated for 2 d with SAC was not altered by rIL-4, which suggests that rIL-4 did not exert its inhibitory activity on rIL-2-induced B cell proliferation by enhancing rIL-2-mediated differentiation . Delayed addition of a neutralizing anti-IL-4 antiserum demonstrated that a period of contact of at least 24 h between IL-4 and B cell blasts was necessary for the development of the antagonistic effect of IL-4 on IL-2-mediated growth of activated B cells . These data demonstrate that IL-4 antagonizes the B cell growth-promoting effect of IL-2 without affecting the differentiation of preactivated B cells in response to IL-2.

J Exp Med, 1988 Oct 1, 168(4), 1225 - 35
The Bb fragment of complement factor B acts as a B cell growth factor; Peters MG et al.; The process of B cell growth and differentiation into plasma cells is highly regulated and may be influenced by a large number of inflammatory mediators, including complement components . We have studied the regulatory influence of Bb, a 60-kD peptide created during the cleavage of complement Factor B by Factor D and C3b . Purified Bb alone had no effect on proliferation and differentiation of human splenic or tonsillar B cells . However, when B cells were activated by Staphylococcus aureus Cowan I (SAC), Bb enhanced proliferation in a dose-dependent manner . Bb also enhanced proliferation when cocultured with SAC and suboptimal concentrations of purified 60-kD B cell growth factor (HMW-BCGF), a previously described lymphokine that is known to possess growth-promoting activity . However, Bb had no effect on cells treated with optimal concentrations of HMW-BCGF . Like HMW-BCGF, Bb's major effect was on the larger in vivo activated B cells . Half-maximal enhancement of proliferation was reached at a Bb concentration of 1-10 nM . Of note is the fact that antibody to Factor B recognized HMW-BCGF, and an mAb to HMW-BCGF also recognized Factor B and Bb, but not Ba . Moreover, radiolabeled Bb bound saturably to activated B cells and to an EBV-transformed human B cell line . The binding of Bb was inhibited by HMW-BCGF but not by Ba or IgG . Thus, Bb is antigenically and functionally related to HMW-BCGF, and can act as a B cell growth and differentiation factor at potentially physiologic concentrations . These data suggest that Bb may be important in amplifying the immune response in areas of inflammation . Since complement activation occurs at inflammatory sites long before induction of HMW-BCGF synthesis, Bb may be an early signal for the clonal expansion of antigen-activated B cells.

Clin Immunol Immunopathol, 1988 Oct, 49(1), 159 - 65
Differential effect of anti-HLA class I monoclonal antibodies on resting and in vivo-induced B lymphocytes; de la Sen ML et al.; The role of HLA class I antigens in B cell triggering was investigated by analyzing the effect of monoclonal antibodies (MAbs) directed to such molecules on the in vitro function of resting and in vivo-induced lymphoblastoid (LB) B cells . Staphylococcus aureus Cowan I (SAC)-induced proliferation of high-density B lymphocytes was markedly inhibited by W6/32 MAb (reactive with a monomorphic determinant contributed by an alpha-chain and beta 2-microglobulin) but not by FG2/2 MAb (reactive with beta 2-microglobulin) . The inhibition was not due to either a toxic effect or a change in the response kinetics . In contrast, LB B cells' spontaneous DNA synthesis and IgG production was not altered by such MAb, although these cells possessed surface HLA class I antigens . These findings suggest that HLA class I molecules are involved in the activation process of resting but not mature B lymphocytes.

Biochem Genet, 1988 Oct, 26(9-10), 543 - 55
Purification and characterization of cytoplasmic creatine kinase isozymes of Xenopus laevis; Robert J et al.; The soluble creatine kinase isozymes CK-II, CK-III, and CK-IV from Xenopus laevis have been purified to apparent homogeneity and their subunits characterized by means of molecular weight, peptide pattern, and dissociation-reassociation experiments . CK-III and CK-IV are homodimeric isozymes whose subunits are distinct in both molecular weight (42,000 and 41,000, respectively) and Staphylococcus aureus V8 peptide pattern . In dissociation-reassociation experiments, those two subunits do form active heterodimeric isozymes with one another or with rabbit M-CK subunits . Hybrid CK-III/IV isozymes occur also during embryonic differentiation and in adult heart muscle, whereas most other adult tissues contain only homodimeric CK-III or CK-IV isozymes . The CK-II isozyme is a heterodimer composed of one CK-III subunit and another subunit specific to CK-II (Mr = 41,000) . Neither in vivo nor in vitro does this subunit seem able to form homodimers or heterodimers with CK-IV and rabbit M-CK subunits . If we take into account the apparent association of CK-I isozyme with cellular organelles, these results corroborate earlier statements and suggest that the CK isozyme system of X . laevis is encoded by at least four differentially regulated genomic loci.

Immunol Lett, 1988 Oct, 19(2), 127 - 32
Effect of serine proteinase from Staphylococcus aureus on in vitro stimulation of human lymphocytes; Prokesova L et al.; In a broad concentration range (0.1-100 micrograms/ml) the serine proteinase (SP) from Staphylococcus aureus has no cytotoxic effect on human peripheral blood lymphocytes and does not stimulate them in culture . However, it affects the action of a number of polyclonal activators . In a concentration of 100 micrograms/ml SP completely eliminates blastic transformation after stimulation with B cell mitogens (NDCM, S . aureus and Escherichia coli), lowers the blastic transformation after stimulation with PWM and SPA, and does not affect the blastic transformation after stimulation with PHA . SP (100 micrograms/ml) reduces the concentration of Ig in stimulated cultures (stimulation with PWM, NDCM, S . aureus and E . coli) far below the Ig level of unstimulated controls . This effect can be ascribed to an influence on cell stimulation, not to the proteolytic cleavage of secreted Ig, although SP can partially digest Ig . The effect on lymphocyte stimulation takes place when the SP is added to the culture together with the mitogen, or 18 h after the mitogen . This implies that SP does not affect the first stage of stimulation.

Chemioterapia, 1988 Oct, 7(5), 306 - 8
In vitro activity of sulbactam/ampicillin and ampicillin against methicillin-sensitive and methicillin-resistant Staphylococcus aureus and Staphylococcus epidermidis; Fabbri A et al.; Minimum inhibitory concentrations (MICs) of ampicillin and ampicillin + sulbactam (1:1) against 165 strains of Staphylococcus aureus and 72 strains of Staphylococcus epidermidis have been evaluated . The activity of the combination was very good . A concentration of 16 micrograms/ml + 16 micrograms/ml inhibited 96.9% of S . aureus and the 100% of S . epidermidis strains (at the same concentration ampicillin alone inhibited only 55.15% and 56.9% of S . aureus and S . epidermidis strains respectively) . Activity against methicillin-resistant S . aureus (14.5%) was poor, whereas against methicillin-resistant S . epidermidis (67.2%) the combination maintained high efficacy.

Zh Mikrobiol Epidemiol Immunobiol, 1988 Oct, (10), 22 - 4
{Effect of the ABO blood group phenotype on the Staphylococcus aureus bacterial carrier state}; Veselov AIa et al.; 326 employees of 4 medical institutions (1 regional hospital, 2 city hospitals and a maternity clinic) were examined for the presence of S . aureus carriership . Examinations were made every 3 months for 3 recent years . The results of these examinations were compared with the distribution of the blood groups in the AB0 system among the carriers . Constant and malignant carrier state was detected mainly in persons with blood group A.

Appl Environ Microbiol, 1988 Oct, 54(10), 2583 - 5
Is free halogen necessary for disinfection?
Williams DE, Elder ED, Worley SD.
The principle of Le Chatelier was used in demonstrating that 3-chloro-4,4-dimethyl-2-oxazolidinone (compound 1) itself kills Staphylococcus aureus rather than the very small amount of free chlorine in hydrolysis equilibrium with compound 1 . On the other hand, when the N-bromo analog of compound 1 (compound 1B) was used as the disinfectant, the mixture of combined compound 1B and free bromine formed in the hydrolysis equilibrium provided disinfection . When the hydrolysis equilibrium for 1B was suppressed to the level at which a negligible amount of free bromine remained in solution, combined compound 1B was much more efficacious than combined compound 1 at killing S . aureus.

Appl Environ Microbiol, 1988 Oct, 54(10), 2486 - 91
Outermost-cell-surface changes in an encapsulated strain of Staphylococcus aureus after preservation by freeze-drying; Ohtomo T et al.; The effects of drying time during freeze-drying on the outermost cell surface of an encapsulated strain of Staphylococcus aureus S-7 (Smith, diffuse) were investigated, with special attention paid to capsule and slime production . To quantify capsule and slime production, capsule antigen production and cellular characteristics such as growth type in serum-soft agar, cell volume index, and clumping factor reaction were examined . After freeze-drying the colonial morphology of strain S-7 was altered from a diffuse to a compact type in serum-soft agar . In accordance with these changes, the titer of the clumping factor reaction increased while the cell volume index, capsule and slime production, and capsule antigen production were markedly decreased in parallel with the period of freeze-drying . The ability of the strain to adhere to collagen, fibrinogen, and soybean lectin was also compared before and after freeze-drying . Fibrinogen levels slightly increased when 10% skim milk and 2% honey were used as cryoprotective agents and showed a remarkable increase when 0.05 M phosphate buffer was used as a control . Also, the ability of strain S-7 to adhere to soybean lectin declined, whereas no changes were observed for collagen under any conditions . Strain S-7 was phage nontypable before freeze-drying but the number of typable cells increased after freeze-drying; phage-typable cells reacted to phage 52 alone after 5 h of freeze-drying, but additional cells also proved to be phage typable to phage 42E after 10 h . Electron micrographs indicated that strain S-7, an encapsulated strain, was converted to an unencapsulated state after freeze-drying.(ABSTRACT TRUNCATED AT 250 WORDS)

Can J Vet Res, 1988 Oct, 52(4), 445 - 50
The use of liposomally-entrapped gentamicin in the treatment of bovine Staphylococcus aureus mastitis; MacLeod DL et al.; The effect of incorporation of gentamicin in liposomes on intracellular killing of Staphylococcus aureus was studied in vitro in cultured bovine mammary macrophages, and in experimental bovine mastitis . Liposomes were prepared by reverse-phase evaporation and ranged in size from 0.1 to 1.0 micron in diameter (mean 0.51 micron), with an encapsulation efficiency of gentamicin of 27.4% . Liposomes were taken up by in vitro cultured macrophages but intracellular killing of S . aureus over 12 h was not significantly enhanced when treatment with liposomally-entrapped gentamicin was compared to free gentamicin . Treatment of experimentally-induced S . aureus mastitis in five lactating Holstein cows (20 quarters) failed to show significant differences in bacterial counts when treatment with liposomally-entrapped gentamicin was compared to treatment with free gentamicin or blank liposomes plus free gentamicin . Gentamicin concentrations exceeded the in vitro determined minimum inhibitory concentration for 48 h when quarters were treated with 50 mg gentamicin on two occasions 24 h apart.

Am J Vet Res, 1988 Oct, 49(10), 1681 - 7
Serum and synovial fluid steady-state concentrations of trimethoprim and sulfadiazine in horses with experimentally induced infectious arthritis; Bertone AL et al.; The tarsocrural joints of 11 horses were inoculated with 1.2 to 2.16 x 10(6) viable Staphylococcus aureus organisms susceptible to a trimethoprim-sulfadiazine (TMP-SDZ) combination with minimal inhibitory concentration (MIC) of 0.25 micrograms of TMP/ml and 4.75 micrograms of SDZ/ml . Antimicrobial treatment consisted of oral administration of a TMP-SDZ combination--30 mg/kg of body weight given once daily (group-1 horses) or 60 mg/kg given as 30 mg/kg every 12 hours (group-2 horses) . Paired serum and synovial fluid samples were obtained before intra-articular inoculation with the S aureus, after inoculation with S aureus but before antimicrobial treatment, and after inoculation at various hourly intervals after oral administration of the TMP-SDZ combination . The TMP-SDZ combination was administered daily in the 2 dosages for 21 days . Samples were collected after day 3 of repetitive drug administration so that drug steady-state concentration would have been achieved . Serum and synovial fluid samples were analyzed for TMP and SDZ concentrations . Administration of the TMP-SDZ combination at a dosage of 30 mg/kg once daily was not effective in maintaining TMP or SDZ concentrations above the MIC of TMP-SDZ for the S aureus (0.25 and 4.75 micrograms/ml for TMP and SDZ, respectively) in the infected synovial fluid or in maintaining adequate TMP concentration in the serum . The alternative use of the TMP-SDZ combination at a dosage of 60 mg/kg given as 30 mg/kg every 12 hours was effective in maintaining serum and synovial fluid concentrations of TMP and SDZ that were greater than the MIC for the infective organism.(ABSTRACT TRUNCATED AT 250 WORDS)

J Clin Microbiol, 1988 Oct, 26(10), 2187 - 90
Evaluation of various antibiotics for induction of L forms from Staphylococcus aureus strains isolated from bovine mastitis; Owens WE; Forty-five strains of Staphylococcus aureus were treated with 11 antibiotics and tested for induction to L forms . Thirty-seven strains were induced to L forms with at least one antibiotic, while eight strains produced no L forms under the conditions used . L forms were induced only with beta-lactam antibiotics and with a combination of penicillin and streptomycin . Novobiocin induced no true L forms but induced intermediate forms from seven strains . Strains resistant to penicillin yielded L forms only with beta-lactam antibiotics active against penicillin-resistant organisms . Strains failing to yield L forms were susceptible to the antibiotics used, and resistance appeared to play no role in the induction of L forms with these strains.

J Bone Joint Surg Am, 1988 Oct, 70(9), 1383 - 92
A histological study of acute hematogenous osteomyelitis following physeal injuries in rabbits; Whalen JL et al.; In young rabbits, the effects of an intravenous injection of Staphylococcus aureus alone, and in combination with a traumatic injury of the proximal tibial physis, were studied by light and electron microscopy . Metaphyseal osteomyelitis and radiographic changes were seen within forty-eight hours after the injury in all rabbits that had a growth-plate disruption and bacteremia . An intravenous injection of bacteria alone produced no morphological or microbiological evidence of infection . In the absence of trauma, normal tibiae were sterile after forty-eight hours . Foreign-body particles have been shown to accumulate in the fine vessels that are adjacent to the growth plate, but we found no similar deposition of bacteria or evidence of phagocytic removal in this area . Phagocytosis of bacteria by neutrophils did not appear to be impaired in the distal third of the metaphysis, but a delayed inflammatory response that allowed proliferation of bacteria and destruction of tissue was observed in the proximal two-thirds of the metaphysis after trauma.

J Bone Joint Surg Am, 1988 Oct, 70(9), 1341 - 7
The prevention of infection in open fractures . An experimental study of the effect of antibiotic therapy; Worlock P et al.; Using an experimental rabbit model of a contaminated open fracture of the tibia that was fixed with an intramedullary pin, we assessed the effect of a single dose of cephradine in preventing post-traumatic osteomyelitis in which the infecting organism was Staphylococcus aureus . We paid particular attention to the effect of a delay in giving the antibiotic . The frequency of osteomyelitis in the animals in a control group (no antibiotic) was 91 per cent . When a single injection of cephradine was given one hour before inoculation with the bacteria, the rate was 30 per cent, a statistically significant reduction (p less than 0.01) . When cephradine was not administered until one to four hours after inoculation with the bacteria, the average rate of osteomyelitis was 51 per cent, a 40 per cent reduction compared with the rate for the control group . The effect of the antibiotic therefore persisted even when the initial dose was delayed for four hours after bacterial inoculation.

Ann Surg, 1988 Oct, 208(4), 451 - 9
More is better . Antibiotic management after hemorrhagic shock; Livingston DH et al.; Previous reports suggest that standard antibiotic prophylaxis is ineffective in reducing the incidence of wound infection after hemorrhagic shock . This study investigated the use of larger and longer doses of antibiotic in a model of staphylococcal infection after hemorrhagic shock . Sprague-Dawley rats resuscitated from hemorrhagic shock were injected with either 10(6), 10(8) or 10(10) Staphylococcus aureus subcutaneously . Five treatments were investigated: 1) control (no antibiotic), 2) short-course cefazolin (CEF) (SHORT), 30 mg/kg intraperitoneal (IP), 30 minutes before and 4 hours after inoculation, 3) long-course CEF (LONG), 30 mg/kg IP, 30 minutes before and 4 hours after inoculation, and thereafter, every 8 hours for 3 days, 4) mega-CEF (MEGA) 200 mg/kg IP, 30 minutes before and 4 hours after inoculation, and 5) mega-long CEF (MEGA-LONG), 200 mg/kg IP, 30 minutes before and 4 hours after inoculation, and thereafter, every 8 hours for 3 days . Abscess number, weight, and diameter were measured on Day 7 . At the 10(6) inoculum, SHORT was effective in both shocked and unshocked animals . In the 10(10) group, all antibiotic regimens decreased the 100% mortality that followed shock without treatment, but they had little effect on abscess formation . In unshocked rats at the 10(8) inoculum, SHORT was effective in reducing abscess number, diameter, and weight (all p less than 0.05 vs . control) . After hemorrhagic shock, SHORT did not decrease abscess frequency, but it did diminish abscess diameter . LONG significantly decreased abscess diameter and abscess weight (both p less than 0.05) . After shock, both MEGA and MEGA-LONG reduced abscess number (p less than 0.05 vs . control) and MEGA-LONG was superior to all other regimens at the 10(8) inoculum . These experimental data show that increasing both the dose and duration of antibiotic administration is more effective than standard short-course antibiotic prophylaxis in preventing experimental infection after hemorrhagic shock.

J Antibiot (Tokyo), 1988 Oct, 41(10), 1374 - 94
Synthesis and structure-activity relationships in the cefpirome series . I . 7-{2-(2-Aminothiazol-4-yl)-2-(Z)-oxyiminoacetamido}-3-{(su bstituted-1-pyridinio)methyl}ceph-3-em-4-carboxylate s; Lattrell R et al.; 7-{2-(2-Aminothiazol-4-yl)-2-(Z)-oxyiminoacetamido}-3-{(s ubs tituted-1-pyridinio)methyl}ceph-3-em-4-carboxylates II are a group of beta-lactam antibiotics with extraordinary high antibacterial activity . The promising member of this group, cefpirome (HR 810, II-1) is a candidate for clinical use . Synthetic pathways to II starting from cefotaxime derivatives I or 7-aminocephalosporanic acid (7-ACA) are described . A preferred method for the conversion of I to II or 7-ACA to precursors III respectively employs iodotrimethylsilane and an excess of the pyridine base . Structure-activity studies reveal an optimum overall activity in the series of pyridines with fused saturated and unsaturated rings or cyclopropyl- and alkoxy substituents . Favorable oxyimino substituents are methyl, ethyl, difluoromethyl and carbamoylmethyl groups . Acidic substituents lead to decreased activity against Staphylococcus aureus SG 511 . Introduction of halogen in the thiazole nucleus causes improvement of activity against the K1 beta-lactamase producing Klebsiella aerogenes 1082 E strain.

J Allergy Clin Immunol, 1988 Oct, 82(4), 535 - 43
Leukotriene C4 generation from human eosinophils stimulated with IgG-Aspergillus fumigatus antigen immune complexes; Cromwell O et al.; Sepharose beads coated with IgG stimulate eosinophils to produce leukotriene C4 (LTC4) . This observation has been extended with specific immobilized IgG/antigen immune complexes to elicit mediator generation . An extract of Aspergillus fumigatus was covalently coupled to Sepharose beads and incubated with the IgG fraction of immune serum from patients with allergic bronchopulmonary aspergillosis . These beads elicited generation of 7.72 +/- 1.7 pmol of LTC4 immunoreactive material (n = 5) from 1 X 10(6) normal eosinophils of greater than 86% purity, and significantly less LTC4 (0.73 +/- 0.19 pmol per 10(6) cells; n = 3) was produced by eosinophils after incubation with beads treated with IgG from normal nonimmune serum . The maximum antibody-dependent release achieved represented approximately 20% of that induced by the calcium ionophore (A23187) . LTC4 was measured by radioimmunoassay and validated by reverse-phase high-performance liquid chromatography . The amount of LTC4 generated was dependent on the concentration of A . fumigatus-specific IgG, and mediator release was completely abolished by prior adsorption of the IgG fraction onto Sepharose-protein A (Staphylococcus aureus) . Grass pollen-specific IgG antibody/antigen complexes, in combination with Sepharose beads, also triggered generation of LTC4 immunoreactive material . There was no evidence to suggest that IgE/A . fumigatus immune complexes triggered LTC4 generation, although IgE myeloma protein, in association with Sepharose beads, was a weak stimulus . The efficacy of the IgG immune complex-dependent stimulation of eosinophils suggests a possible physiologic mechanism whereby these cells could participate in the inflammatory changes associated with allergic bronchopulmonary aspergillosis and similar allergic disorders.

Jpn J Antibiot, 1988 Oct, 41(10), 1517 - 37
{Clinical evaluation of cefpodoxime proxetil in the treatment of skin and soft tissue infections . A double blind comparison of cefpodoxime proxetil and cefaclor}; Yura J et al.; In order to objectively evaluate the effectiveness, safety and usefulness of the new oral cephem cefpodoxime proxetil (CS-807, CPDX-PR) for the treatment of skin and soft tissue infections, a double-blind comparative study was undertaken using cefaclor (CCL) as the control drug . CPDX-PR and CCL were administered for 7 days at daily doses of 400 mg (divided into 2 portions) and 750 mg (divided into 3 portions), respectively . A total of 243 patients (118 in the CPDX-PR group and 125 in the CCL group) was treated in this study . The effectiveness, safety and usefulness were evaluated in 222 (106 in the CPDX-PR group and 116 in the CCL group), 234 (113 in the CPDX-PR group and 121 in the CCL group) and in 223 patients (107 in the CPDX-PR group and 116 in the CCL group), respectively . There were no differences in patients' backgrounds between the 2 groups, except for the presence or the absence of surgical treatments . The results we obtained are summarized below: 1 . In the evaluation of clinical efficacy by the subcommittee, excellent, good, fair and poor efficacy were observed in 36, 43, 17 and 10 patients in the CPDX-PR group, respectively; the efficacy rate was, therefore, calculated to be 74.5% . As for the CCL group, respective results were observed in 50, 39, 17 and 10 patients, indicating an efficacy rate of 76.7% . There was no significant difference between the 2 groups . Improvement rates judged by physicians in charge were 80.2% in the CPDX-PR group and 88.8% in the CCL group . Moreover, no significant difference in diseases or severity were found between the 2 groups . 2 . As for the bacteriological efficacy, the 2 groups showed high elimination rates, as 90.1% and 91.6% of the disease causing bacteria were eliminated in the CPDX-PR group and in the CCL group, respectively . Elimination rates in single infections with Staphylococcus aureus were determined to be 85.7% in the CPDX-PR group and 85.0% in the CCL group . 3 . Although 6 patients in the CPDX-PR group and 2 patients in the CCL group developed side effects, which were mainly gastrointestinal symptoms, there was no significant difference in the incidence of side effects between the 2 groups . Abnormal laboratory values were found in 5 patients in the CPDX-PR group and 1 patient in the CCL group . 4 . There was no significant difference in the usefulness between the 2 groups.(ABSTRACT TRUNCATED AT 400 WORDS)

Pathol Biol (Paris), 1988 Oct, 36(8), 956 - 8
{Course of the resistance of Staphylococcus aureus to pefloxacin . A study based on 782 strains isolated in 1985 and 1986}; Jean-Pierre H et al.; We studied the evolution of the resistance of Staphylococcus aureus to pefloxacin during 1985 and 1986 in eight hospital units . The pefloxacin resistant Staphylococcus aureus, which amounted to 8.9% in 1985 reached 27% in 1986 . They are found mainly among the methicillin-resistant Staphylococcus . There is no obvious relationship between this resistance and the consumption of pefloxacin in each of the units.

J Laryngol Otol, 1988 Oct, 102(10), 877 - 82
Septic cavernous and lateral sinus thrombosis: modern diagnostic and therapeutic principles; Tveteras K et al.; The incidence of both lateral and cavernous sinus thrombophlebitis has been significantly reduced in the antibiotic era . Since septic cavernous sinus thrombosis (CST) is mainly a complication of facial abscesses and septic lateral sinus thrombosis (LST) is almost invariably due to chronic otitis media, both conditions are of clinical relevance to the otolaryngologist . The predominant bacterium in septic CST is Staphylococcus aureus whereas in septic LST the bacteriology is very similar to that found in chronic otitis media . The diagnosis of septic CST can be established in most cases after thorough clinical examination, and contrast computerized tomography (CT) using the coronal projection usually confirms the clinical diagnosis . The signs and clinical course of septic LST are non-specific and the final diagnosis rests upon radiological investigations including CT-scan . The treatment of both conditions consists of broad-spectrum antibiotics, including beta-lactamase resistant penicillin in cases of septic CST . Most cases of septic LST also require surgical intervention . Two cases of septic intracranial sinus thrombosis are presented . The need for early diagnosis and treatment of this potentially lethal condition is emphasized.

J Infect Dis, 1988 Oct, 158(4), 702 - 9
Implications of acquired oxacillin resistance in the management and control of Staphylococcus aureus infections; Massanari RM et al.; Refinements in testing for resistance to penicillinase-resistant penicillins (PRP) in Staphylococcus aureus have resulted in confusion in classifying isolates as PRP susceptible or resistant . Specifically, a group of organisms has been identified that produce large amounts of beta-lactamase and appear borderline resistant . These organisms have been called "occult resistant" or "acquired oxacillin-resistant" S . aureus (AORSA) . A retrospective study was conducted to evaluate the implication of this in vitro phenomenon in managing patients with AORSA infections . Among 134 patients with S . aureus infections, 89 were infected with oxacillin-susceptible S . aureus (OSSA), 26 with AORSA, and 19 with oxacillin-resistant S . aureus (ORSA) . There were no significant differences in outcomes when OSSA and AORSA infections were treated with PRP (chi 2MH = .990; P = .32) . These results do not suppor the contention that AORSA infections should be managed differently from OSSA infections . Identifying AORSA may not be helpful in guiding antimicrobial therapy or predicting the outcome of infections with AORSA.

Jpn Circ J, 1988 Oct, 52(10), 1191 - 200
Isolation and characterization of two beta-type cardiac myosin in the canine atrium; Komuro I et al.; Recently, we demonstrated that more beta-type myosin heavy chain (HC) was expressed in the overloaded atrium, and that there were 2 structurally different beta-type myosin heavy chains in the bovine heart . To determine the existence of the 2 beta-type HC in other animals and to clarify the characteristics of these beta-type HCs, we produced tricuspid regurgitation and pulmonary stenosis in the canine heart, and performed an immunological study using 3 monoclonal antibodies, 2 beta-type specific antibodies (HMC14 and 50) and 1 alpha-type specific antibody (CMA19) . In an immunohistochemical study, serial cryostat sections revealed that some myofibers reacted with HMC50 (HC beta 2), but almost no fibers were labeled with HMC14 in the normal atrium . However, in overloaded atria, not only HC beta 2 but the HC, reacted with HMC14 (HC beta 1) . By affinity chromatography, HC beta 2 was fractionated from normal atrial myosin using HMC50 and HC beta 1 was fractionated from overloaded atrial myosin using HMC14 . These 2 HC beta's were subjected to digestion by alpha-chymotrypsin, staphylococcus aureus V8 protease, and cyanogen bromide, and proved to have different peptide fragments . In respect to enzymatic properties, the Ca2+-activated ATPase activities of HC beta 1 and beta 2 were almost the same but lower than that of HC alpha . We concluded that the isozymic transition of HC alpha to HC beta in the atrium was experimentally induced by hemodynamic overload and that HC beta 1, which was hardly recognized in the normal atrium but highly induced by overload, was structurally different from HC beta 2, as expressed in the normal atrium.

Appl Environ Microbiol . 1988 Oct;54(10):2590.
Stabilities of lyophilized Staphylococcus aureus typing bacteriophages; Zierdt CH; Staphylococcus aureus bacteriophages (25 phages) were lyophilized in aliquots 12 to 18 years ago and stored in vacuo at -20 degrees C . Eight viruses each lost one log titer, while seventeen retained the original titers . The use of lyophilized phages provided more reproducible phage typing and reduced by 75% the complexity and cost . This important test is thus made feasible for more laboratories.

J Am Acad Dermatol, 1988 Oct, 19(4), 673 - 8
Staphylococcus aureus colonization of burrows in erythrodermic Norwegian scabies . A case study of iatrogenic contagion; Shelley WB et al.; Scanning electron microscopy demonstrated extensive bacterial colonization of scabies burrows honeycombing the stratum corneum of an elderly woman with erythroderma . Cultures of scybala revealed hemolytic Staphylococcus aureus, possibly responsible for the erythroderma . Epidemiologic data revealed a trail of scabies through two nursing homes and one hospital during the 2-year period that physicians believed she had a drug-induced erythroderma.

J Clin Endocrinol Metab, 1988 Oct, 67(4), 749 - 54
In vitro induction of anti-thyroid microsomal antibody-secreting cells in peripheral blood mononuclear cells from normal subjects; Iitaka M et al.; Secretion of immunoglobulin G (IgG) and IgM antithyroid microsomal antibodies (AMA) was induced in vitro by coculturing non-T cells (B lymphocytes) and autologous CD4 (helper/inducer) cells from normal subjects stimulated with pokeweed mitogen (PWM) or a combination of human thyroid microsomal antigen (McAg) and Staphylococcus aureus Cowan I (SAC) strain . With PWM stimulation, AMA production was induced in more IgM-secreting cells (AMA-M) than IgG-secreting cells (AMA-G) . However, McAg plus SAC stimulation resulted in similar numbers of AMA-G- and AMA-M-secreting cells . PWM induced a significantly greater number of both AMA-M (and generalized IgM)-secreting cells than did McAg plus SAC, while the number of AMA-G-secreting cells induced by the two stimuli were similar . There were no significant differences between autologous or allogeneic CD4 cells from normal subjects or patients with autoimmune thyroid disease (AITD) when cocultured with B cells from normal subjects in terms of helper activity in the induction of AMA-M- or IgM-secreting cells with PWM stimulation . However, with McAg plus SAC, CD4 cells from patients with AITD induced a significantly greater number of AMA-M-secreting cells than did autologous or allogeneic CD4 cells from normal subjects . There was no difference in helper activity between autologous and allogeneic normal CD4 cells in the induction of generalized IgM-secreting cells regardless of the stimulus used . Normal autologous or allogeneic CD8 (suppressor/cytotoxic) cells cocultured with normal B cells and autologous CD4 cells suppressed the induction of AMA-M-secreting cells by PWM stimulation . On the other hand, CD8 cells from patients with AITD suppressed the induction of AMA-M-secreting cells significantly less effectively . All CD8 cells suppressed the induction of IgM-secreting cells equally well . We conclude that 1) B lymphocytes from normal subjects are capable of producing autoantibodies in vitro in the presence of CD4 cells; 2) the helper activity of CD4 cells from patients with AITD to induce AMA-M secreting cells is greater than that of normal CD4 cells with thyroid antigen stimulation; and 3) this helper activity may be due to relatively impaired suppressor activity in thyroid antigen-specific CD8 cells from patients with AITD, whereas the immunoregulatory function of CD8 cells from normal subjects appears to play an important role in the maintenance of self-tolerance.

J Hosp Infect, 1988 Oct, 12(3), 225 - 33
Simple peroperative antimicrobial chemoprophylaxis in elective neurosurgical operations; Ingham HR et al.; From August 1981 to February 1982 postoperative infections due to different strains of penicillin-resistant Staphylococcus aureus occurred in 20 of 467 patients (4.3%) undergoing elective cranial and spinal operations . These infections were not attributable to defects in procedures or the theatre environment, therefore chemoprophylaxis was instituted . In the following 8 months, when patients were given penicillin G and sulphadiazine for 5 days commencing immediately postoperatively, S . aureus infections occurred in five of 579 patients (0.9%) . In a subsequent randomized uncontrolled study, infections occurred in six of 265 patients receiving penicillin (2.3%), three of 270 receiving penicillin and sulphadiazine (1.1%) and one of 45 receiving erythromycin (2.2%) immediately postoperatively for 5 days . In a further study in which 587 patients received penicillin for 5 days commencing immediately preoperatively, infections due to S . aureus occurred in six (1.1%) . Infections due to gram-negative organisms were seen in five (0.4%) of 1167 patients in the two uncontrolled studies.

J Hosp Infect, 1988 Oct, 12(3), 151 - 62
Colonization priority among Staphylococcus aureus strains--correlation with phage-type; Rosdahl VT et al.; We have studied the distribution of phage-type patterns among strains of Staphylococcus aureus isolated from patients in a burns unit . From 51 patients the same phage-type was isolated from succeeding swabs during the observation period . In 20 patients new types were introduced, but the original strain remained . In 23 patients the first strain was replaced by one other strain, in eight patients two or more . Strains of type 95 seemed to have a high colonization priority, whereas strains of group III had a low one . In 1986 phage-typing was performed on two or more S . aureus strains from the same patient, in 4561 instances . Recurrence of strains of the same phage-type pattern was demonstrated in 70% of the patients when the first and the fourth sample were compared . The "newer epidemic" strains of phage-type 95 and of the 94,96 complex had the highest percentage of recurrence (more than 80%) when adjacent samples were compared, and 68-69% when the first and the fourth sample were compared . The good colonization capacity of these strains might be one of the explanations why they occur frequently today although they are resistant only to penicillin.

Cell Immunol, 1988 Oct 1, 116(1), 195 - 215
Induction of suppressor cells to T- and B-cell proliferative responses and immunoglobulin production by monoclonal antibodies recognizing the CD3 T-cell differentiation antigen; Kunicka JE et al.; Monoclonal antibodies (mAb's) recognizing the CD3 T-cell differentiation antigen induced the generation of suppressor cells . These cells inhibited (1) proliferative responses of human peripheral blood mononuclear cells (PBMC) to PHA and allogeneic cells in mixed leukocyte culture; (2) proliferative responses of purified E-rosette-negative cells to Staphylococcus aureus Cowans I; and (3) de novo immunoglobulin synthesis and secretion in the pokeweed mitogen (PWM)-induced differentiation system . Monoclonal antibodies recognizing other T-cell differentiation antigens (anti-Leu 2a, anti-Leu 3a, and anti-Leu 5) did not induce the generation of suppressor cells, even at very high antibody concentrations . Statistically significant differences were not observed in the ability of the OKT3 and anti-Leu 4 mAb's to induce suppressor cells . Monocytes were not required for the generation of anti-CD3-induced suppressor cells . F(ab')2 fragments of the OKT3 mAb's were equally effective when compared with intact antibody molecules in inducing suppressor cells, although they did not induce proliferative responses . Proliferation was not required for the induction of suppressor cells . Irradiation (2500 rad) of PBMC before incubation with the anti-CD3 mAb did not affect the generation of suppressor cells . Furthermore, anti-CD3-induced suppressor cells were radioresistant . Addition of recombinant IL-2 to the cultures of responding cells and suppressor cells did not reverse the suppression . In vitro treatment of anti-CD3-induced suppressor cells with either the OKT4 mAb plus complement or the OKT8 mAb plus complement partially decreased the suppression of proliferative responses of PBMC to PHA or allogeneic cells in mixed lymphocytes culture . However, treatment with both OKT4 and OKT8 mAb's plus complement or the OKT11 mAb plus complement completely abolished the suppression . These results suggest that the suppressor cells are of the T11+T4+T8- and T11+T4-T8+ phenotypes . In other experiments, T4+T8- and T8+T4- cells were isolated from PBMC treated for 48 hr with anti-CD3 mAbs . Both these two populations significantly inhibited proliferative responses of autologous PBMC to PHA and de novo immunoglobulin synthesis and secretion by mixtures of purified T4 and B cells from normal donors, in the PWM-induced differentiation system . These results demonstrate that anti-CD3-induced suppressor cells are of the T4 or T8 phenotype . Treatment of purified T4+T8- and T8+T4- cells with anti-CD3 mAb's resulted in the generation of suppressor cells, suggesting that the precursors of the anti-CD3-induced suppressor cells can belong to either of these two populations.(ABSTRACT TRUNCATED AT 400 WORDS)

J Clin Lab Immunol, 1988 Oct, 27(2), 97 - 102
Myeloperoxidase secretion during phagocytosis: a case of a patient with impaired bactericidal activity; Edwards SW et al.; We describe a case of a 5-year-old male patient with prolonged and extensive osteomyelitis of the left femur . Staphylococcus aureus was grown from blood cultures taken upon admission and also from pus drained from an incised hip joint . A defect in immune function was suspected and neutrophil function was assessed . Chemotaxis and phagocytosis were normal, but phagocytosed S . aureus were not killed as efficiently as in control neutrophils . No inherent defect in the ability of these neutrophils to generate reactive oxidants was observed, but an unusual luminol-dependent chemiluminescence response was obtained during phagocytosis of latex beads or opsonized S . aureus: This was characterized by an initial rapid, but transient increase occurring within 1 min of addition of phagocytic stimulus . Whereas during phagocytosis of latex beads by control neutrophils less than 1% of the total myeloperoxidase activity was detected extracellularly, up to 15% was released from the patient's neutrophils . We propose that release of myeloperoxidase from the patient's neutrophils during phagocytosis reduces the intraphagosomal concentration of this enzyme and thus impairs the efficiency of intracellular killing of S . aureus.

Inflammation, 1988 Oct, 12(5), 515 - 24
Thiourea and dimethylthiourea decrease human neutrophil bactericidal function in vitro; Jackson JH et al.; Addition of thiourea (TU) or dimethylthiourea (DMTU) decreased killing of Staphylococcus aureus, 502A, and decreased concentrations of hydrogen peroxide (H2O2), and hydroxyl radical (.OH), but not superoxide anion (O2-.) or lysozyme concentrations, in mixtures containing human neutrophils in vitro . Addition of TU or DMTU also decreased concentrations of H2O2, .OH, or hypochlorous acid (HOCl) in neutrophil-free mixtures exposed to beta-D-glucose and glucose oxidase, gamma irradiation, or HOCl, respectively . Our results suggest that TU or DMTU can decrease neutrophil-mediated killing of bacteria by inhibiting O2 metabolite-dependent bactericidal mechanisms.

Inflammation, 1988 Oct, 12(5), 447 - 53
Variable effect of toxic shock toxins from different sources on neutrophil function in vitro; Berger EM et al.; Toxic shock syndrome toxins (TSST) are 23-30 kD proteins that have been isolated from incubation media of strains of Staphylococcus aureus cultured from patients with toxic shock syndrome (TSS) . Injection of TSST into animals produces many of the symptoms that characterize TSS including shock, fever, and multiple organ failure . We found that addition of increasing concentrations of TSST-1-VP1035-16A, but not TSST-PEC, TSST-SEC, staphylococcal enterotoxin A or B, progressively decreased human neutrophil bactericidal activity against S . aureus, 502A in vitro . TSST-1-VP1035-16A, but not the other toxins, also decreased superoxide anion and hydrogen peroxide concentrations in mixtures containing neutrophils and phorbol myristate acetate (PMA) in vitro . The results indicate that various preparations of TSST have different effects on neutrophil function in vitro and, accordingly, may have different effects in other in vitro and in vivo models of TSS.

J Med Microbiol, 1988 Oct, 27(2), 117 - 23
Characterisation of methicillin-resistant Staphylococcus aureus isolates by restriction endonuclease digestion of chromosomal DNA; Jordens JZ et al.; Clinical isolates of Staphylococcus aureus from the London Hospital were characterised by genetic analysis of antibiotic-resistance determinants and by restriction endonuclease digestion of chromosomal DNA and compared with isolates from elsewhere in the UK . Restriction enzyme digestion of chromosomal DNA confirmed that a single strain of methicillin-resistant S . aureus (MRSA) persists at the London Hospital, although its antibiotic-resistance profile and plasmid carriage are not constant . Methicillin-sensitive isolates, on the other hand, each had readily distinguishable and unique DNA restriction patterns . The DNA restriction digest pattern of the London Hospital MRSA isolates was identical to that of "epidemic" (E) MRSA isolates from the Thames regions . By contrast, other MRSA isolates had DNA restriction patterns which differed from those of EMRSA isolates and from each other . These results confirm the discriminatory value of restriction pattern analysis as a typing method.

Infect Immun, 1988 Oct, 56(10), 2702 - 8
Association of toxic shock toxin-1 determinant with a heterologous insertion at multiple loci in the Staphylococcus aureus chromosome; Chu MC et al.; Most Staphylococcus aureus strains associated with toxic shock syndrome and producing toxic shock syndrome toxin 1 (TSST-1) require tryptophan because of a genetic defect in tryptophan biosynthesis . The association between TSST-1 production and tryptophan auxotrophy was not correlated with the phage type, the colonization site, or the disease status of the patient from whom the isolate came . Protoplast fusion and transformation mapping located the genetic determinant of TSST-1 production (tst) very close to the trp operon in such strains and very close to tyrB in a Trp+ TSST-1+ strain . Southern blot hybridization of ClaI-restricted chromosomal DNA with a tst-specific probe revealed a common homologous segment in all of the Trp+ strains with tst linked to tyrB . These results confirmed that the tst determinant in Trp- strains is located at one site, whereas in Trp+ TSST-1+ strains the determinant is located elsewhere on the S . aureus chromosome . It is suggested that the TSST-1 determinant is associated with the insertion of a transposonlike segment into several sites on the S . aureus chromosome.

Gen Physiol Biophys, 1988 Oct, 7(5), 467 - 73
The structure of Staphylococcus aureus alpha-toxin-induced ionic channel; Krasilnikov OV et al.; Polyethylene glycols (PEG) with molecular weight less than or equal to 3000 were shown to effectively protect human erythrocytes from osmotic lysis induced by alpha-staphylotoxin (ST) . PEG with MW less than 3000 do not change the conductivity of ion channels induced by ST in bilayer lipid membranes (BLM) . Changing the bilayer from a pure phosphatidylcholine (PC) to a negatively charged phosphatidylserine (PS) film results in an asymmetry of the current-voltage characteristics . This is evidenced by the asymmetrical position of the ST-channel pore in bilayer membranes . The results obtained allow to conclude that the ST-channel is an interprotein pore filled with water (with an inner diameter of 2.5-3 nm and a length of approximately 10 nm) . It is composed of six molecules of alpha-toxin from Staphylococcus aureus . The ST-channel incorporates into a membrane with only one mouth in contact with the polar lipid heads and the other one protruding 4.5-5 nm from the bilayer plane in water solution.

Biochem J, 1988 Sep 15, 254(3), 919 - 22
Accumulation of acyl-enzyme intermediates during turnover of penicillins by the class A beta-lactamase of Staphylococcus aureus PC1; Pratt RF et al.; The interaction of dansylpenicillin with the class A Staphylococcus aureus PCI beta-lactamase yielded an accumulating intermediate with fluorescence enhanced beyond that of the substrate . Acid quenching of the reaction mixture yielded a denatured enzyme with 1 molar equivalent of dansyl group covalently bound to it . A similar quenching experiment with the PC1 beta-lactamase and {14C}benzylpenicillin yielded an enzyme with 1 molar equivalent of 14C covalently bound . These data indicate that in turnover of S-type penicillins by the PC1 beta-lactamase deacylation is rate-determining . This has not indicate previously been demonstrated for a class A beta-lactamase.

J Am Vet Med Assoc, 1988 Sep 15, 193(6), 683 - 6
Management of a severely comminuted fracture of the third metacarpal bone in a horse; Orsini JA et al.; A 4-year-old Standardbred stallion sustained a severely comminuted fracture involving the second, third, and fourth metacarpal bones . The fracture was repaired using two 14-hole broad dynamic compression plates positioned at 90 degrees to one another, allowing one plate to protect the other in the bending mode . An autologous cancellous bone graft obtained from the tuber coxae was added at the site of the defect in the mid- to upper third of the third metacarpal bone . Complications associated with the fixation included a Staphylococcus aureus infection 5 months after surgery, laminitis that developed in the opposite forelimb 6 months after the fracture, and septic tenosynovitis in the contralateral hind limb . The septic tenosynovitis prolonged hospitalization for a total of 20 months . Both postoperative problems resolved when the stallion returned to breeding use.

J Immunol, 1988 Sep 15, 141(6), 2072 - 8
A novel IFN-gamma regulated human melanoma associated antigen gp33-38 defined by monoclonal antibody Me14/D12 . I . Identification and immunochemical characterization; Giuffre L et al.; A novel melanoma-associated differentiation Ag whose surface expression can be enhanced or induced by IFN-gamma was identified by mAb Me14/D12 . Testing of numerous tumor cell lines and tumor tissue sections showed that Me14/D12-defined Ag was present not only on melanoma but also on other tumor lines of neuroectodermal origin such as gliomas and neuroblastomas and on some lymphoblastic B cell lines, on monocytes and macrophages . Immunoprecipitation by mAb Me14/D12 of lysates from {35S}methionine-labeled melanoma cells analyzed by SDS-PAGE revealed two polypeptide chains of 33 and 38 KDa, both under reducing and nonreducing conditions . Cross-linking experiments indicated that the two chains were present at the cell surface as a dimeric structure . Two-dimensional gel electrophoresis showed that the two chains of 33 and 38 KDa had isoelectric points of 6.2 and 5.7, respectively . Treatment of the melanoma cells with tunicamycin, an inhibitor of N-linked glycosylation, resulted in a reduction of the Mr from 33 to 24 KDa and from 38 to 26 KDa . Peptide maps obtained after Staphylococcus aureus V8 protease digestion showed no shared peptides between the two chains . Although biochemical data indicate that Me14/D12 molecules do not correspond to any known MHC class II Ag, their dimeric structure, tissue distribution, and regulation of IFN-gamma suggest that they could represent a new member of the MHC class II family.

J Immunol, 1988 Sep 15, 141(6), 2000 - 5
Human recombinant IL-4 induces activated B lymphocytes to produce IgG and IgM; Defrance T et al.; In this report, we describe a novel biologic activity of IL-4 namely, its ability to induce activated human B cells to produce IgM . Staphylococcus aureus Cowan I-activated blasts prepared from high density tonsil B cells were found to secrete IgG and IgM, but no IgE, when cultured in the presence of rIL-4 . The differentiating activity of rIL-4 was totally blocked by a neutralizing anti-IL-4 antiserum, therefore demonstrating that the IgG/IgM-inducing activity of rIL-4 was an intrinsic property of IL-4 . rIL-4 was only minimally inducing Ig production of blasts prepared from low density B cells, whereas it induced B cell blasts prepared from high density B cells to secrete a high amount of Ig . Delayed additions of the neutralizing anti-IL-4 antiserum demonstrated that a 48-h contact between IL-4 and B cell blasts was required for optimal Ig production . The IL-4-mediated IgG and IgM production was neither suppressed by IFN-gamma nor by anti-CD23 mAb 25, whereas these agents have been shown earlier to inhibit IgE production of enriched B cells cultured in the presence of IL-4 . These data indicate that the IgG/IgM-inducing activity of IL-4 is not regulated like the IL-4-induced IgE production by enriched B cells.

Biochem J, 1988 Sep 15, 254(3), 765 - 71
Resynthesis of sphingomyelin from plasma-membrane phosphatidylcholine in BHK cells treated with Staphylococcus aureus sphingomyelinase; Allan D et al.; About 60-65% of the total sphingomyelin in intact BHK cells is in a readily accessible pool which is rapidly degraded by Staphylococcus aureus sphingomyelinase . No more sphingomyelin is broken down in cells which have been fixed with glutaraldehyde or lysed with streptolysin O, suggesting that all the sphingomyelin which is available to the enzyme is on the cell surface . The inaccessible pool of sphingomyelin does not equilibrate with the plasma-membrane pool, even after prolonged incubation . Experiments using {3H}-choline show that much more phosphocholine is released from the intact cells treated with sphingomyelinase than can be accounted for by breakdown of the original cell-surface pool of sphingomyelin; the excess appears to be a consequence of the breakdown of sphingomyelin newly resynthesized at the expense of a pool of phosphatidylcholine which represents about 8% of total cell phosphatidylcholine and may reside in the plasma membrane . This would be consistent with resynthesis of cell-surface sphingomyelin by the phosphatidylcholine: ceramide phosphocholinetransferase pathway, which has previously been shown to be localized in the plasma membrane . However, in {3H}palmitate-labelled cells there appeared to be no accumulation of the diacylglycerol expected to be produced by this reaction, and no enhanced synthesis of phosphatidate or phosphatidylinositol; instead there was an increased synthesis of triacylglycerol . A similar increase in labelling of triacylglycerol was seen in enzyme-treated cells where the sphingomyelinase was subsequently removed, allowing resynthesis of sphingomyelin which occurred at a rate of about 25% of total sphingomyelin/h . Treatment of BHK cells with sphingomyelinase caused no change in the rates of fluid-phase endocytosis or exocytosis as measured with {3H}inulin.

J Mol Biol, 1988 Sep 5, 203(1), 183 - 95
Structure of fatty acid synthetase from the Harderian gland of guinea pig . Proteolytic dissection and electron microscopic studies; Kitamoto T et al.; Limited proteolysis and electron microscopic observation of fatty acid synthetase from the Harderian gland of guinea pig was performed to elucidate the higher-order structures of this multifunctional protein . Staphylococcus aureus V8 protease dissected the 250,000 Mr subunit of fatty acid synthetase into 120,000, 70,000, 35,000 and 30,000 Mr fragments, which were aligned in this order from the NH2 terminus . Some of the protease-resistant fragments produced with elastase, trypsin and lysyl endopeptidase were purified and fragment-specific antibodies (A40L, A33E and A25T) were prepared . A25T and A33F specifically bound the 35,000 and 30,000 Mr fragments, and A40L recognized the region between the 120,000 and 70,000 Mr fragments . Electron microscopic studies employing rotary shadowing, unidirectional shadowing and negative staining revealed that the overall dimension of the enzyme was 22 nm x 15 nm x 7 nm, and that two elongated subunits mainly composed of three subregions were in contact with each other at a few, three at most, points with two holes between them . The outer two attachment sites were often not in contact, indicating a certain flexibility of subunits at their ends . Immunocomplexes composed of fatty acid synthetase and fragment-specific antibodies were isolated and observed under the electron microscope . The attachment sites of A40L and A33E were located at the end of the minor and the major axes of the ellipsoidal contour of the molecule, respectively . Based on these results, the three-dimensional structure of animal fatty acid synthetase is discussed.

S Afr Med J, 1988 Sep 3, 74(5), 223 - 4
Methicillin-resistant Staphylococcus aureus at Tygerberg Hospital; Peddie EF et al.; During 1985 Staphylococcus aureus was isolated from blood cultures of 74 patients at Tygerberg Hospital who were suffering from serious illness compatible with systemic spread of the organism . Twenty-six isolates (35%) were community-acquired and none were methicillin-resistant, while 48 were hospital-acquired of which 23 (48%) were methicillin-resistant . Methicillin resistance appears to be a problem confined to hospital isolates of S . aureus.

Proc Natl Acad Sci U S A, 1988 Sep, 85(18), 7008 - 12
Sequencing of proteins from two-dimensional gels by using in situ digestion and transfer of peptides to polyvinylidene difluoride membranes: application to proteins associated with sensitization in Aplysia; Kennedy TE et al.; We have developed a method for obtaining partial internal amino acid sequence data from proteins isolated directly from preparative two-dimensional polyacrylamide gels . Proteins from a crude cell homogenate are separated using preparative two-dimensional polyacrylamide gel electrophoresis . Then, the gel is stained with Coomassie blue and the protein spots of interest are cut out . The in situ protein is digested with Staphylococcus aureus V8 protease in a second polyacrylamide gel and the peptides are separated by one-dimensional polyacrylamide gel electrophoresis . The peptides are then electroblotted onto a polyvinylidene difluoride membrane, visualized using Coomassie blue, cut out, and sequenced using an automated gas phase sequencer . Using this method, we have obtained amino acid sequence data for two proteins that are altered after long-term sensitization: actin and Aplysia protein 407 . In addition, we have obtained amino acid sequence data for rat protein 425, a protein that appears to be homologous to Aplysia protein 407.

Clin Orthop, 1988 Sep, (234), 211 - 6
Brodie's abscess . A long-term review; Stephens MM et al.; In 20 patients with 21 Brodie's abscesses, a long-term review revealed that 13 occurred in the second decade of life . All had local symptoms for six weeks or more . The tibia was involved in 11 cases and seven of these were in the proximal metaphysis . The erythrocyte sedimentation rate (ESR) was elevated in only six cases . When the ESR was more than 40 mm per hour, recurrence was more likely . Staphylococcus aureus was cultured from 11 abscesses . Curettage and antibiotics for six weeks were adequate for treatment in most cases . However, lesions larger than 3 cm in diameter should be grafted, and patients with an elevated ESR require more aggressive decompression and prolonged antibiotic therapy . Lesions within the neck of the femur pose particular anatomic problems and should not be approached laterally . All cases were followed to full bone maturity . No significant leg length inequality was clinically or roentgenologically apparent . If an abscess was juxtaphyseal, deformity of the epiphysis could develop.

Arch Otolaryngol Head Neck Surg, 1988 Sep, 114(9), 1000 - 2
Osteomyelitis of the clavicle; Alessi DM et al.; Osteomyelitis of the clavicle is a rare entity and can occur as a complication of head and neck surgery . Ten consecutive cases of the clavicular osteomyelitis were reviewed at the University of California Medical Center, Los Angeles, over the past seven years . Six cases were associated with prior surgical procedures, and five cases presented as chronic wound drainage . One case was related to a pharyngocutaneous fistula following a supraglottic laryngectomy . Four patients presented with acute symptoms resulting from hematogenous spread, and two of the four patients had Staphylococcus aureus on blood cultures . Long-term intravenous antibiotic therapy (six to eight weeks) was used to successfully treat cases of hematogenously spread osteomyelitis . Wide surgical debridement was the mainstay of treatment in the chronic conditions, with antibiotic therapy having a secondary role . Myocutaneous flaps were required in two patients who had had surgery and antecedent radiotherapy . To conclude, the surgeon should be aware that osteomyelitis of the clavicle can occur as a complication of head and neck procedures . In addition, the treatment of the chronic form of clavicular osteomyelitis is surgical debridement and possible flap reconstruction.

Plast Reconstr Surg, 1988 Sep, 82(3), 480 - 5
Muscle flaps in the management of vascular grafts in contaminated wounds: an experimental study in dogs; Cruz NI et al.; This study evaluates in an animal model the efficacy of muscle flaps in protecting the fabric vascular prosthesis when placed in contaminated wounds . A total of 20 adult mongrel dogs received a 2-cm interpositional polytetrafluoroethylene (PTFE) graft to each femoral artery at the groin level . During the surgical procedure, the wounds were inoculated with a Staphylococcus aureus suspension containing either 1 x 10(4) or 1 x 10(5) organisms per milliliter . In half the animals, the grafts were wrapped with a distally based sartorius muscle flap before a standard two-layer closure was completed . One month after the surgery, all the animals were sacrificed and quantitative cultures were performed of the grafts and wounds . The muscle flaps were capable of protecting the vascular prosthesis with inoculums up to 1 x 10(4) organisms (p less than 0.05), but at greater bacterial contamination their efficacy was no longer significant.

Z Naturforsch {C}, 1988 Sep-Oct, 43(9-10), 656 - 64
On the relationships between molecular conformation, affinity towards penicillin-binding proteins, and biological activity of penicillin G-sulfoxide; Beise F et al.; The binding capacity of penicillin G-sulfoxide towards the penicillin-binding proteins (PBP) of Staphylococcus aureus H was studied . The sulfoxide and its parent compound, penicillin G, differ only in two aspects, the sulfur-bound oxygen and an altered conformation of the five-membered thiazolidine-ring system . These minor alterations of the penicillin structure resulted in a drastical decrease of binding activity (about two orders of magnitude) of the sulfoxide derivative towards its target enzymes . Furthermore, the sulfoxide did not exhibit the selectivity of subinhibitory doses for PBP 3, as could be observed for penicillin G . The biological consequences of this behaviour were monitored via growth curves, uptake of cell wall label, and analysis of the cell wall . Binding studies revealed that comparable growth inhibition and impairment of cell wall label uptake were achieved by at least a 100-fold higher penicillin G-sulfoxide concentration, compared to its parent compound . In cell wall analysis, the application of high doses of the antibiotics, i.e . nearly saturated PBP, verified the above mentioned observation . Surprisingly, small but significant differences in cell wall composition occurred using subinhibitory doses, probably due to the altered affinity towards PBP 3, supporting the hypothesis of an important role of this PBP in peptidoglycan transpeptidation.

Jpn J Antibiot, 1988 Sep, 41(9), 1205 - 11
{In vitro susceptibilities of clinical isolates of Staphylococcus aureus}; Igari J et al.; In vitro susceptibilities of 350 strains of Staphylococcus aureus to ampicillin (ABPC), methicillin (DMPPC), cloxacillin (MCIPC), cefazolin (CEZ), cefmetazole (CMZ), cefmenoxime (CMX), latamoxef (LMOX) and 5 non-beta-lactam antimicrobial agents were determined according to the standard method of Japan Society of Chemotherapy . Frequencies of the appearance of resistant organisms (MIC greater than or equal to 12.5 micrograms/ml) to beta-lactam antibiotics were 45% for ABPC, 27% for DMPPC, 11% for MCIPC, 24% for CEZ, 15% for CMZ, 36% for CMX and 51% for LMOX . To non-beta-lactam antimicrobial agents, resistant strains appeared at 31% to gentamicin, 15% to amikacin, 0.6% to minocycline (MINO), 1% to norfloxacin (NFLX) and 65% to fosfomycin (FOM) . More than 80% of DMPPC-resistant strains were also resistant to LMOX, CMX, ABPC, FOM, and CEZ, but most of those were susceptible to MINO and NFLX . Incidence of DMPPC-CEZ resistant S . aureus was 23% of the 350 strains tested . As stated above, multiply resistant strains of S . aureus are present throughout different hospitals in Okinawa.

Jpn J Surg, 1988 Sep, 18(5), 576 - 9
Double valve replacement for infective endocarditis in a hemodialysis patient: a case report; Kinoshita K et al.; A 57-year-old man who was in end stage renal failure underwent an aortic and mitral valve replacement for the progression of cardiac dysfunction, secondary to Staphylococcus aureus infective endocarditis . Cardiac surgery was performed using a Hemo-Concentrator during cardiopulmonary bypass, 82 months following the initiation of hemodialysis . This is the second report in the literature of a successful double valve replacement for infective endocarditis and congestive heart failure in a chronic hemodialysis patient.

Am J Vet Res, 1988 Sep, 49(9), 1494 - 6
Prevalence and characterization of Staphylococcus aureus in young goats; de Centorbi ON et al.; Thirty-six Staphylococcus aureus isolates recovered from 35 of 204 young goats at slaughter were characterized . All isolates were susceptible to cephalothin, clindamycin, chloramphenicol, gentamicin, kanamycin, and amikacin . All but 2 were susceptible to erythromycin and tetracycline, and 19 and 20 were susceptible to penicillin and ampicillin, respectively . Thirteen isolates were classified as biotype A, 9 isolates were classified as biotype B, 8 isolates were classified as biotype C, and 6 isolates were classified as intermediate between B and C or were not biotypable . Six biotype A isolates were enterotoxigenic; 4 produced enterotoxin B, 1 produced enterotoxin C, and 1 produced enterotoxin D . Two biotype B strains produced enterotoxin B, and all 8 biotype C isolates produced enterotoxin C and the toxic shock syndrome toxin-1.

Drug Intell Clin Pharm, 1988 Sep, 22(9), 695 - 6
Oxacillin-associated hypokalemia; Schlaeffer F; Antibiotic-induced hypokalemia does not occur frequently, but has been described with aminoglycosides, amphotericin B, and ureido penicillins . A patient with Staphylococcus aureus bacteremia who developed severe hypokalemia while on high doses of oxacillin is presented . To our knowledge this is the first reported case of oxacillin-associated hypokalemia.

Can J Microbiol, 1988 Sep, 34(9), 1050 - 7
Molecular analyses of conjugative, gentamicin-resistance plasmids from staphylococcal clinical isolates; Zorbas I et al.; Gentamicin-resistant Staphylococcus aureus and Staphylococcus epidermidis strains which were isolated from infants with staphylococcal bacteremia were analyzed for the presence of self-transmissible gentamicin-resistance (Gmr) plasmids . Conjugative GMr plasmids of approximately 43.8-63 kilobases (kb) were found in all S . aureus strains . Inter- and intra-species transfer of Gmr plasmids by conjugation was observed from S . aureus to S . aureus and to S . epidermidis recipient strains . However, neither inter- nor intra-species transfer of gentamicin resistance by conjugation was observed with nine out of nine S . epidermidis donor strains which were mated with either S . epidermidis or S . aureus recipient strains . These conjugative Gmr plasmids were unable to comobilize a smaller (15-kb) plasmid present in all but two S . aureus clinical isolates . Many of the conjugative Gmr plasmids also carried genetic determinants for kanamycin, tobramycin, neomycin, and ethidium bromide resistance, and for beta-lactamase synthesis . EcoRI restriction endonuclease digests of the S . aureus Gmr conjugative plasmids revealed three different digestion patterns . Four EcoRI restriction endonuclease digestion fragments of 15, 11.4, 6.3, and 4.6 kb in size were common to all plasmids . These plasmids and conjugative Gmr staphylococcal plasmids from other geographical regions shared restriction digestion fragments of similar molecular weights . DNA hybridization with biotinylated S . aureus plasmid pIZ7814 DNA revealed a high degree of homology among these plasmids . A 50.9-kb plasmid from one of the nonconjugative S . epidermidis clinical isolates showed homology with the probe DNA but lacked a portion of a 6.3-kb fragment which was present in all conjugative plasmids and believed to carry much genetic information for conjugation.

Spine, 1988 Sep, 13(9), 1042 - 8
A comparison between magnetic resonance imaging and scintigraphic bone imaging in the diagnosis of disc space infection in an animal model; Szypryt EP et al.; In a controlled prospective study, 33 rabbits were used to compare the efficacy of magnetic resonance imaging (MRI) to scintigraphy in diagnosing pyogenic infection of the intervertebral disc . A suitable inoculum of Staphylococcus aureus (10(8) organisms) was injected into a test level while a similar volume of sterile culture medium was injected into a control disc in each animal . Plain radiographs, MRI, and scintigraphic bone images then were performed at regular intervals after operation . The imaging studies were interpreted blind, as was the final histologic assessment . Five animals died of respiratory complications following sedation . In the remaining 28 animals, 23 developed infection at the test level, and four developed infection at the control disc . Magnetic resonance imaging was found to be more sensitive than scintigraphy in diagnosing discitis, particularly in the early stages of the disease . The overall results showed MRI had a sensitivity of 93%, specificity of 97%, and accuracy of 95% . There were two false-negative results and one false-positive . In contrast, scintigraphy had a sensitivity of 41%, specificity of 93%, and accuracy of 68% . There were two false-positive and 16 false-negative results . Plain radiographs had a sensitivity of 82%, specificity of 93%, and accuracy of 88% . There were two false-positive and five false-negative results . The MRI appearance of discitis was characteristic from an early stage in the disease, and preceded the changes on scintigraphy and plain radiographs by several days in the majority of cases.

J Clin Microbiol, 1988 Sep, 26(9), 1810 - 3
Sensitive immune dot blot test for diagnosis of Chlamydia trachomatis infection; Mearns G et al.; The sensitivity and specificity of an immune dot blot test (IDBT), which relies on a 125I-labeled genus-specific monoclonal antibody to detect the Chlamydia lipopolysaccharide (LPS) antigen, were improved by pretreatment of specimens with proteinase K . This enzyme destroys protein A and therefore eliminates false-positive reactions caused by the presence of Staphylococcus aureus . Proteinase K treatment also improved the ability of the assay to detect the Chlamydia LPS antigen . When the improved IDBT was compared with culture for detection of C . trachomatis in 1,394 urogenital specimens obtained from a genitourinary medicine clinic, the overall sensitivity was 96%, and LPS antigen was detected in 76 of 83 (92%) specimens that yielded less than 10 inclusions in culture . The specificity and positive and negative predictive values of the test were 97, 81.5, and 99%, respectively . Of 123 conjunctival swabs, 7 were positive by both tests and 4 swabs were positive only by IDBT . This improved IDBT provides a simple, reliable alternative to culture for the detection of C . trachomatis in urogenital and conjunctival specimens.

Biochem J, 1988 Sep 1, 254(2), 419 - 26
Characterization of {3H}palmitate- and {3H}ethanolamine-labelled proteins in the multicellular parasitic trematode Schistosoma mansoni; Wiest PM et al.; Biosynthetic labelling experiments with cercariae and schistosomula of the multicellular parasitic trematode Schistosoma mansoni were performed to determine whether {3H}palmitate or {3H}ethanolamine was incorporated into proteins . Parasites incorporated {3H}palmitate into numerous proteins, as judged by SDS/polyacrylamide-gel electrophoresis and fluorography . The radiolabel was resistant to extraction with chloroform, but sensitive to alkaline hydrolysis, indicating the presence of an ester bond . Further investigation of the major 22 kDa {3H}palmitate-labelled species showed that the label could be recovered in a Pronase fragment which bound detergent and had an apparent molecular mass of 1200 Da as determined by gel filtration on Sephadex LH-20 . Schistosomula incubated with {3H}ethanolamine for up to 24 h incorporated this precursor into several proteins; labelled Pronase fragments recovered from the three most intensely labelled proteins were hydrophilic and had a molecular mass of approx . 200 Da . Furthermore, reductive methylation of such fragments showed that the {3H}ethanolamine bears a free amino group, indicating the lack of an amide linkage . We also evaluated the effect of phosphatidylinositol-specific phospholipase C from Staphylococcus aureus: {3H}palmitate-labelled proteins of schistosomula and surface-iodinated proteins were resistant to hydrolysis with this enzyme . In conclusion, {3H}palmitate and {3H}ethanolamine are incorporated into distinct proteins of cercariae and schistosomula which do not bear glycophospholipid anchors . The {3H}ethanolamine-labelled proteins represent a novel variety of protein modification.

J Appl Physiol, 1988 Sep, 65(3), 1372 - 6
Bacterial adherence to human endothelial cells; Thomas PD et al.; The adult respiratory distress syndrome (ARDS) is frequently caused by exposure of the lung endothelium to circulating endotoxin (lipopolysaccharide, LPS) and pulmonary infections frequently develop during the course of ARDS . The present studies demonstrate that LPS and interleukin 1 (IL-1, a mediator released by endothelial cells after exposure to LPS) enhance the adherence of Staphylococcus aureus to human umbilical vein endothelial cells . gamma-Interferon, another mediator that induces expression of some cell surface antigens on endothelial cells, had no effect on bacterial adherence . The adherence of bacteria to endothelium was increased by prior opsonization of the bacteria with fresh human serum and was reduced by prior absorption of the serum with bacteria before the use of the serum for opsonization . The capacity of LPS to increase bacterial adherence was time dependent and was maximally expressed after 6 h of exposure; it was blocked by exposure of endothelial cells to LPS in the presence of reduced temperature or dactinomycin (Actinomycin D) . These observations suggest that circulating LPS not only can trigger the development of ARDS but also may predispose the lung to the development of pulmonary infections by increasing adherence of bacteria to endothelium.

J Antimicrob Chemother, 1988 Sep, 22(3), 377 - 84
Elimination of nasal carriage of methicillin-resistant Staphylococcus aureus with mupirocin during a hospital outbreak; Hill RL et al.; During a hospital outbreak of methicillin-resistant Staphylococcus aureus (MRSA), involving more than 200 patients, 40 patients and 32 hospital staff who were stable nasal carriers of MRSA received topical application of 2% mupirocin, formulated in a white soft paraffin and lanolin ointment, to their anterior nares for five days . Nasal carriage was eliminated in all patients and staff, usually within the first 48 h of treatment . Of the 40 patients, 36 remained clear of nasal MRSA for the duration of their follow-up (mean = 2 weeks) and four became re-colonized one to five weeks after their course . Immediately after the course, the number of patients with MRSA isolated from wounds and wrists fell from 16 to 7, and from 16 to 3, respectively . Of the 32 staff, all were negative one week after the course, and of the 22 still available for follow-up at eight weeks, all were consistently negative (mean period of follow-up = 7.8, range = 1-20 weeks) . Four patients and five staff were re-colonized with MRSA between one to five, and two to twelve weeks, respectively, after treatment . Overall, in the post-treatment follow-up, 98.6% of the staff-weeks and 90.1% of the patient-weeks were free of nasal MRSA . MICs of mupirocin for both pre and post treatment isolates were all 0.03 or 0.06 mg/l . The elimination of nasal MRSA by mupirocin, and the introduction of isolation facilities, were associated with the control of the outbreak.

Zh Mikrobiol Epidemiol Immunobiol, 1988 Sep, (9), 24 - 8
{The growth and protein A accumulation of Staphylococcus aureus A-676 on a medium made from nonfood raw material}; Dubinina GP et al.; In this research the experimental analytical method of balancing culture media, based on the determination of the yield of the target product, i.e . staphylococcal protein A, has been used . The medium, developed in the course of this research work on the basis of a hydrolysate of protein-containing waste products of the tanning industry, makes it possible to achieve the growth of producer strains similar to that achieved in peptone-yeast medium . The parameters of the growth of S . aureus strain A-676 and the biological activity of protein A obtained in the newly developed medium have been studied.

Rev Infect Dis, 1988 Sep-Oct, 10(5), 1035 - 7
Delayed rupture of the radial artery caused by catheter-related sepsis; Arnow PM et al.; Arterial catheter-related sepsis was detected in six adult patients during 45 months by prospective surveillance of blood culture results . In two of the patients, infection of the radial artery caused delayed arterial rupture and either hemorrhage or pseudoaneurysm formation despite prompt catheter removal and antibiotic treatment . Features present in these two cases but not in the four uncomplicated cases were infection caused by Staphylococcus aureus and persistence of systemic signs of infection for more than 2 days after catheter removal . A review of the literature showed that these two features have been noted in almost all adult patients previously reported to develop arterial rupture as a complication and not in patients with uncomplicated arterial catheter-related sepsis . Identification of patients at increased risk for this complication may facilitate prompt diagnosis of arterial rupture or exploration and possible resection of an infected segment of artery before rupture occurs.

J Antimicrob Chemother, 1988 Sep, 22 Suppl C, 159 - 66
Use of ofloxacin in open fractures and in the treatment of post-traumatic osteomyelitis; Ketterl R et al.; In two open prospective studies, the efficacy and tolerance of ofloxacin in the prevention of infection in patients with open fractures (n = 58) and in the treatment of chronic post-traumatic osteomyelitis (n = 115) were examined . In the study with open fractures, bone and/or soft tissue infection occurred in only four cases (6.5%) . During an observation period of at least 12 months, post-traumatic osteomyelitis was seen in two patients with III degree open fractures (9%), while in the groups with I degree and II degree open fractures no bone infection could be found . Therefore, the rate of post-traumatic osteomyelitis related to all patients was 3.3% . In the second study with 115 patients suffering from chronic post-traumatic osteomyelitis 141 different Gram-positive and Gram-negative pathogens were isolated . 73% were Gram-positive cocci with Staphylococcus aureus in more than 50% of the cases . An elimination rate of more than 90% was found in the Gram-positive and Gram-negative bacteria, leading to a clinical cure in 85% and a recurrence of infection in 5% of the cases . The tolerability of ofloxacin was excellent . No drug-related allergic reactions were observed . Diarrhoea and headache occurred in less than 2% of patients . With adequate surgical treatment, ofloxacin proved to be a useful antimicrobial agent in the prevention and therapy of bone infection.

Proc Soc Exp Biol Med, 1988 Sep, 188(4), 444 - 50
Role of prostaglandins in controlling plasma fibronectin levels; Cheng CY et al.; Fibronectin is a normal glycoprotein component of plasma, interstitial fluid, and extracellular matrix which has binding sites for collagen, gelatin, actin, glycosaminoglycans, fibrin, Staphylococcus aureus, and some cells . Since it is a dimer, it can crosslink these substances to each other or to extracellular components of basement membrane, thereby affecting many physiological processes . The level of circulating fibronectin is markedly reduced following even moderate blunt or operative trauma, thermal injury, starvation, advanced cancers, hemorrhage, etc . Replacement therapy has been tried with some success in patients who become septic following multiple injuries . The reduction in plasma fibronectin has been attributed to several causes including consumption by binding to cell debris at the site of injury, binding to circulating cell debris and its subsequent removal by elements of the phagocytic system, and degradation by proteolytic cleavage . However, the amount of fibronectin removed from circulation raises some question about this . In this paper, we used indomethacin, ibuprofen, imidazole, and essential fatty acid deprivation to inhibit the synthesis of prostaglandins in young adult rats . Thirty minutes after ip administration of one of the inhibitors, the rats were subjected to a midline laparotomy and mild intestinal manipulation . Blood samples were taken at intervals following closure of the incision and analyzed for fibronectin . In all cases, the normal decline in plasma fibronectin seen in untreated rats was abrogated by inhibiting prostaglandin synthesis . Since imidazole specifically inhibits thromboxane A synthesis, this strongly suggests that thromboxanes directly or indirectly control the trauma-induced reduction in circulating fibronectin . This was confirmed by ip injection of thromboxane into the rats which resulted in a decline in plasma fibronectin levels.

Ann Thorac Surg, 1988 Sep, 46(3), 353 - 5
Acute purulent mediastinitis and sternal osteomyelitis after closed chest cardiopulmonary resuscitation: a case report and review of the literature; Mensah GA et al.; Numerous complications have been associated with cardiopulmonary resuscitation . Acute purulent staphylococcal mediastinitis and sternal osteomyelitis are, however, unusual and do not appear to have been reported previously in association with closed chest resuscitation . Sternal fracture during chest compressions and subsequent hematogenous seeding of the resultant retrosternal hematoma with Staphylococcus aureus led to purulent mediastinitis and sternal osteomyelitis in our patient . The source of bacteremia may have been a resolving phlebitis at an intravenous catheter insertion site . Early diagnosis, aggressive surgical debridement, and antibiotic therapy were key to a successful outcome.

J Biochem (Tokyo), 1988 Sep, 104(3), 375 - 82
The amino acid sequence of ribonuclease N1, a guanine-specific ribonuclease from the fungus Neurospora crassa; Takahashi K; The complete amino acid sequence of ribonuclease N1 (RNase N1), a guanine-specific ribonuclease from a fungus, Neurospora crassa, was determined by conventional protein sequencing, using peptide fragments obtained by tryptic digestion of cyanogen bromide-treated RNase N1 and by Staphylococcus aureus V8 protease digestion of heat-denatured RNase N1 . The results showed that the protein is composed of a single polypeptide chain of 104 amino acid residues cross-linked by two disulfide bonds and has a molecular weight of 11,174: (sequence; see text) (Disulfide bonds: C2-C10, C6-C103) The amino acid sequence was homologous with those of RNase T1 (65% identity) and related microbial RNases.

Proc Natl Acad Sci U S A, 1988 Sep, 85(17), 6543 - 6
95- and 25-kDa fragments of the human immunodeficiency virus envelope glycoprotein gp120 bind to the CD4 receptor; Nygren A et al.; 125I-labeled gp120 (120-kDa envelope glycoprotein) from the BH10 isolate of human immunodeficiency virus is cleaved to a limited extent with the glutamate-specific protease from Staphylococcus aureus . After disulfide bond reduction, fragments with approximate molecular masses of 95, 60, 50, and 25 kDa are produced . Tests for binding to CD4-positive cells show that only two fragments, the 95- and 25-kDa peptides, are observed in cleavage products that retain the selective binding capacity of gp120 . Radiosequence analysis of the fragments after sodium dodecyl sulfate/polyacrylamide gel electrophoresis and electroblotting demonstrates that the 95-kDa fragment lacks the N-terminal region of gp120 and starts at position 143 of the mature envelope protein . The 50-kDa fragment starts at the same position . The 25-kDa binding fragment was similarly deduced to be generated as a small fragment from a cleavage site in the C-terminal part of gp120 . The identifications of these fragments demonstrate that radiosequence analysis utilizing 125I-labeled tyrosine residues can function as a useful and reliable method for small-scale determination of cleavage sites in proteins . Combined, the data suggest domain-like subdivisions of gp120, define at least two intervening segments especially sensitive to proteolytic cleavage, and demonstrate the presence of a functional region for receptor binding in the C-terminal part of the molecule.

J Bacteriol, 1988 Sep, 170(9), 4033 - 9
Staphylococcal enterotoxin B gene is associated with a discrete genetic element; Johns MB Jr et al.; The chromosomal location of the enterotoxin B gene in Staphylococcus aureus is unknown . Southern hybridization analysis of the chromosomal DNA from several enterotoxin B (SEB)-producing strains has shown that at least 26.8 kilobases (kb) of DNA is associated with the enterotoxin B gene (entB) . We have found that one end of the entB element is located approximately 1.5 kb downstream of the entB gene . The chromosomal region adjacent to this end of the entB element was found to be homologous in several SEB-producing (SEB+) and SEB-nonproducing (SEB-) S . aureus strains . The chromosomes of all the SEB+ strains studied were homologous for at least 24 kb upstream of the entB gene . Some naturally occurring SEB- strains lacked the entire entB element, while others showed variable homology to the region upstream of the entB gene . These data suggest that the entB gene is part of a discrete genetic element that is at least 26.8 kb in size.

Endocrinology, 1988 Sep, 123(3), 1264 - 73
Enzyme binding-inhibiting assay for iodothyronine 5'-monodeiodinase (5'-MD) and its application to isolation of complementary deoxyribonucleic acid clones for the 5'-MD in rat liver; Boado RJ et al.; To identify and/or quantify type I T4 5'-monodeiodinase (5'-MD) immunologically, polyclonal antibodies were produced by immunization of rabbits with solubilized microsomal proteins (SMP) from rat liver . Pilot studies showed that the antibody binds to, but does not neutralize, rat liver enzyme . We have employed the polyclonal antibody to develop a 5'-MD enzyme binding-inhibiting assay (MBIA) . For this purpose, active, inactive, or synthetic 5'-MD was preincubated with rabbit antibody and removed by Staphylococcus aureus protein-A (Staph-A) . The 5'-MD-binding sites that were left on the Staph-A-bound rabbit antibody were assayed by adding active 5'-MD in fresh liver SMP . After centrifugation of Staph-A, the unbound 5'-MD enzyme activity was measured in the supernatant using {125I}rT3 in the presence of dithiothreitol . Incubation with 3.2 +/- 0.9 micrograms (mean +/- SEM; n = 4) rat liver SMP inhibited the binding of active 5'-MD to protein-A-bound antibody by 50% . Based on doses with similar 50% binding inhibitory activity, liver SMP were 2.3 times more potent than liver microsomes and 19 times more potent than liver homogenate; liver nuclei and mitochondria showed little or no inhibition of the binding of 5'-MD to antibody . The relative 5'-MD content of liver, kidney, pituitary, placenta, and cerebral cortex based on relative potency in the MBIA approximated 100:25:8:3:3 . The threshold of the MBIA approximated 0.9 micrograms liver SMP/tube . The coefficient of variation approximated 2% within an assay and 6% between assays . To obtain a cDNA clone for 5'-MD we used the SMP antibody to screen a rat liver lambda gt11 cDNA expression library . Of 16 positive (antibody-reactive) clones that were isolated, the fusion proteins from only 2 (no . 23 and 54) inhibited the binding of 5'-MD in rat liver SMP to protein-A-bound rabbit antibody in a dose-dependent manner . Western blot analysis showed that the molecular size of liver protein encoded by clones 23 and 54 approximates 31K . Sequence analysis showed that clone 23 insert is 804 basepairs long, contains a single long open-reading frame, and is identical to clone 54 . Southern blot analysis showed that clones 23 and 54 did not cross-hybridize DNA from other SMP antibody-positive clones . Northern blot analysis using clone 23 insert cDNA as the probe showed a hybridization band corresponding to a mRNA approximating 2.8 kilobases.(ABSTRACT TRUNCATED AT 400 WORDS)

Mol Gen Genet, 1988 Sep, 214(1), 108 - 11
Positioning ribosomes on leader mRNA for translational activation of the message of an inducible Staphylococcus aureus cat gene; Dick T et al.; The expression of the chloramphenicol (Cm) - inducible Cm acetyltransferase gene (cat) of the staphylococcal plasmid pUB112 is regulated at the translational level . The leader mRNA preceding the cat coding sequence can form a stable hairpin structure, in which the cat Shine-Dalgarno sequence is masked . Previous work showed that translation of a short leader peptide terminating within the stem of the inhibitory secondary structure is required for basal Cm acetyltransferase (CAT) synthesis and its inducibility . In the present study we shortened this leader peptide by introducing ochre codons in its coding sequence and found that synthesis of the N-terminal part of the leader peptide, terminating directly 5' to the stem, is sufficient to mediate basal and inducible CAT synthesis . Amino acid substitution in this region of the leader peptide abolished inducibility . We suggest that the 5' region of the leader peptide coding sequence specifies a particularly Cm-sensitive translation that represents the Cm-sensor mechanism for cat gene induction.

J Nutr, 1988 Sep, 118(9), 1158 - 64
Decreased amino acid requirements of growing chicks due to immunologic stress; Klasing KC et al.; Experiments were conducted to determine the influence of immunologic stress on methionine and lysine requirements of growing chicks . Immunologic stress was elicited by injection of either Escherichia coli lipopolysaccharide or heat-killed Staphylococcus aureus every other day for 6 d . In the first experiment, diets were formulated to provide methionine levels of 0.30, 0.50 and 0.70% . In the second experiment, diets contained 0.75, 0.90 or 1.2% lysine . In chicks fed amino acid-sufficient diets, those chicks injected with immunogens had slower growth, lower feed intake and poorer efficiency of feed utilization than those injected with saline . The decreases due to immunogens were diminished in chicks fed amino acid-deficient diets . The methionine requirements of saline- and immunogen-injected chicks were above 0.5% and between 0.3 and 0.5%, respectively; the lysine requirements were greater than 0.95% and between 0.7 and 0.95%, respectively . Thus immunogen injection decreased methionine and lysine requirements, probably because of a decreased need of amino acids for growth and tissue accretion . Immunogen-induced depression in serum zinc and increase in serum copper levels were ameliorated by lysine or methionine deficiencies . Compared with saline-injected chicks, immunogen-injected chicks had significantly higher serum interleukin-1 (IL-1) activity by 53% when fed the methionine-sufficient diet, but they did not have significantly greater IL-1 levels when fed the methionine-deficient diet . These observations indicate that the diminished expression of immunologic stress in amino acid-deficient chicks is due to an impaired immune response.

J Bacteriol, 1988 Sep, 170(9), 4365 - 72
Cloning, characterization, and sequencing of an accessory gene regulator (agr) in Staphylococcus aureus; Peng HL et al.; We have previously identified a gene in Staphylococcus aureus, agr, whose activity is required for high-level post-exponential-phase expression of a series of secreted proteins . In this paper, we describe the cloning of this gene in Escherichia coli by using an inserted transposon (Tn551) as a cloning probe . The cloned gene, consisting of a 241-codon open reading frame containing the site of the transposon insertion, was recloned to an S . aureus vector, pSK265, and shown to be functional in S . aureus . Activity was evaluated by determinations of alpha-hemolysin, beta-hemolysin, and toxic shock syndrome toxin-1 production in early-stationary-phase cultures . The cloned gene showed considerable variation with respect to different exoproteins and different host strains compared with the chromosomal agr determinant; this variation could not be attributed to the higher copy number of the cloned gene and probably reflects inapparent subtleties of the regulatory system.

Biochemistry, 1988 Aug 23, 27(17), 6512 - 6
Staphylococcal phosphoenolpyruvate-dependent phosphotransferase system: purification and characterization of the mannitol-specific enzyme IIImtl of Staphylococcus aureus and Staphylococcus carnosus and homology with the enzyme IImtl of Escherichia coli; Reiche B et al.; Enzyme IIImtl is part of the mannitol phosphotransferase system of Staphylococcus aureus and Staphylococcus carnosus and is phosphorylated by phosphoenolpyruvate in a reaction sequence requiring enzyme I (phosphoenolpyruvate-protein phosphotransferase) and the histidine-containing protein HPr . In this paper, we report the isolation of IIImtl from both S . aureus and S . carnosus and the characterization of the active center . After phosphorylation of IIImtl with {32P}PEP, enzyme I, and HPr, the phosphorylated protein was cleaved with endoproteinase Glu(C) . The amino acid sequence of the S . aureus peptide carrying the phosphoryl group was found to be Gln-Val-Val-Ser-Thr-Phe-Met-Gly-Asn-Gly-Leu-Ala-Ile-Pro-His-Gly-Thr-Asp- Asp . The corresponding peptide from S . carnosus shows an equal sequence except that the first residue is Ala instead of Gln . These peptides both contain a single histidyl residue which we assume to carry the phosphoryl group . All proteins of the PTS so far investigated indeed carry the phosphoryl group attached to a histidyl residue . According to sodium dodecyl sulfate gels, the molecular weight of the IIImtl proteins was found to be 15,000 . We have also determined the N-terminal sequence of both proteins . Comparison of the IIImtl peptide sequences and the C-terminal part of the enzyme IImtl of Escherichia coli reveals considerable sequence homology, which supports the suggestion that IImtl of E . coli is a fusion protein of a soluble III protein with a membrane-bound enzyme II . In particular, the homology of the active-center peptide of IIImtl of S . aureus and S . carnosus with the enzyme IImtl of E . coli allows one to predict the N-3 histidine phosphorylation site within the E . coli enzyme.

Biochemistry, 1988 Aug 23, 27(17), 6453 - 7
Drosophila acetylcholinesterase: demonstration of a glycoinositol phospholipid anchor and an endogenous proteolytic cleavage; Haas R et al.; The presence of a glycoinositol phospholipid anchor in Drosophila acetylcholinesterase (AChE) was shown by several criteria . Chemical analysis of highly purified Drosophila AChE demonstrated approximately one residue of inositol per enzyme subunit . Selective cleavage by Staphylococcus aureus phosphatidylinositol-specific phospholipase C (PI-PLC) was tested with Drosophila AChE radiolabeled by the photoactivatable affinity probe 3-(trifluoromethyl)-3-(m-{125I}iodophenyl)diazirine {( 125I}TID), a reagent that specifically labels the lipid moiety of glycoinositol phospholipid-anchored proteins . Digestion with PI-PLC released 75% of this radiolabel from the protein . Gel electrophoresis of Drosophila AChE in sodium dodecyl sulfate indicated prominent 55- and 16-kDa bands and a faint 70-kDa band . The {125I}TID label was localized on the 55-kDa fragment, suggesting that this fragment is the C-terminal portion of the protein . In support of this conclusion, a sensitive microsequencing procedure that involved manual Edman degradation combined with radiomethylation was used to determine residues 2-5 of the 16-kDa fragment . Comparison with the Drosophila AChE cDNA sequence {Hall, L.M.C., & Spierer, P . (1986) EMBO J . 5, 2949-2954} confirmed that the 16-kDa fragment includes the N-terminus of AChE . Furthermore, the position of the N-terminal amino acid of the mature Drosophila AChE is closely homologous to that of Torpedo AChE . The presence of radiomethylatable ethanolamine in both 16- and 55-kDa fragments was also confirmed . Thus, Drosophila AChE may include a second posttranslational modification involving ethanolamine.

Gene, 1988 Aug 15, 68(1), 53 - 62
Expression and secretion of interferon-alpha 1 by Streptomyces lividans: use of staphylokinase signals and amplification of a neo gene; Noack D et al.; A gene coding for mature human interferon, IFN-alpha 1, fused to the expression and secretion signals of a staphylokinase gene (sak) derived from Staphylococcus aureus phage 42D, was inserted into the Streptomyces promoter probe vector pIJ487 . Streptomyces lividans transformed with the recombinant plasmid (pMG341) secreted biologically active IFN-alpha 1 into the culture medium . Expression of the IFN-alpha 1 gene was at least on the translational level directed by the sak signals since numerous upstream stop codons would have prevented the formation of a fusion protein . Long-term continuous chemostat cultivation under various limitation conditions was used to select clones with an IFN-alpha 1 yield increased about 60-100-fold (1-2 x 10(5) IU/ml) . The increase in IFN-alpha 1 formation was accompanied by spontaneous amplification of the adjacent neo gene, but not of the remaining plasmid DNA . Examination of the DNA sequence around the endpoints of the amplified region revealed almost identical stem-loop structures followed by an octanucleotide direct repeat.

APMIS, 1988 Aug, 96(8), 732 - 4
The Tween 80 reaction does not correlate to triglyceride lipase production of Staphylococcus aureus; Rollof J et al.; This comparison of Tween 80 reaction with triglyceride lipase production comprises 357 Staphylococcus aureus strains from various clinical sources . Tween 80 is a water-soluble substrate, and thus estimates esterase activity . Lipase production was assayed using water-insoluble substrates (radiolabelled triglyceride emulsions) . Only a minor proportion of Tween 80-positive strains had significant triglyceride lipase production (greater than 5 mU/10(9) bacteria) . The discrepancy between the two methods was most pronounced in subgroups of samples with low frequency of lipase production, e.g . from isolates from superficial locations such as nasal mucosa and impetigo . Owing to its low specificity, the Tween 80 reaction therefore seems unsuitable as a marker for lipase activity in clinical isolates of Staphylococcus aureus.

Ann Rheum Dis, 1988 Aug, 47(8), 695 - 6
Osteomyelitis presenting as a swollen elbow in a patient with the acquired immune deficiency syndrome; Goh BT et al.; A patient suffering from the acquired immune deficiency syndrome (AIDS), who developed swelling of the left elbow four weeks after Staphylococcus aureus septicaemia is reported . The cause was osteomyelitis of the olecranon process.

Am J Med, 1988 Aug, 85(2), 172 - 6
Staphylococcus aureus bacteremia and recurrent staphylococcal infection in patients with acquired immunodeficiency syndrome and AIDS-related complex; Jacobson MA et al.; PURPOSE: An increased incidence of Staphylococcus aureus bacteremia has recently been described in patients with the acquired immunodeficiency syndrome (AIDS) . However, other risk factors for community-acquired S . aureus bacteremia (including intravenous drug abuse and lymphedema) were present in nearly all these AIDS-related cases of S . aureus infection . Our purpose was to review cases of S . aureus bacteremia that occurred in patients with AIDS or AIDS-related complex (ARC) who did not have a recent history of intravenous drug use, lymphatic obstruction, or neutropenia . PATIENTS AND METHODS: Patients at San Francisco General Hospital between October 1984 and October 1987 with blood culture results positive for S . aureus were identified . A review of this group revealed 22 cases of S . aureus bacteremia that occurred in 18 patients with an underlying diagnosis of AIDS or ARC, none of whom had a recent history of intravenous drug use, lymphedema secondary to Kaposi's sarcoma, or neutropenia . RESULTS: An intravenous catheter was the single most important risk factor for S . aureus bacteremia and was identified as the source for bacteremia in 16 (73 percent) of the 22 episodes . Based on 1986 outpatient clinic records, we calculated an incidence of S . aureus bacteremia occurring in non-intravenous-drug-using male AIDS or ARC patients, 18 to 44 years old, that was 5.4 episodes/1,000 patients . Although the mean duration of appropriate antibiotic therapy was 18 days, late metastatic complications of S . aureus bacteremia occurred in six (35 percent) of 17 AIDS/ARC patients who survived initial antibiotic therapy . CONCLUSION: Non-intravenous-drug-using AIDS and ARC patients (especially those with indwelling venous catheters) appear to be at high risk for S . aureus bacteremia, with a higher late metastatic complication rate than that reported for recent historical control subjects.

Proc Natl Acad Sci U S A, 1988 Aug, 85(15), 5429 - 33
Probing the peptide binding site of the cAMP-dependent protein kinase by using a peptide-based photoaffinity label; Miller WT et al.; A peptide-based photoaffinity label for the catalytic subunit of the cAMP-dependent protein kinase was prepared from the amino acid p-benzoyl-L-phenylalanine {L-Phe(pBz)} . By using solid-phase peptide synthesis methodology, DL-Phe(pBz) was incorporated into the cAMP-dependent protein kinase substrate Leu-Arg-Arg-Ala-Ser-Leu-Gly in place of the phosphorylatable serine . The diastereomeric peptides were separated by reverse-phase HPLC . The peptide substrate analog containing L-Phe(pBz) had a Ki of approximately 110 microM at pH 7.5 . When photolyzed at 350 nm in the presence of the enzyme, this peptide caused time- and concentration-dependent inactivation . Radioactive acetylated L-Phe(pBz) peptide was used to establish the binding stoichiometry of peptide to enzyme; these results, together with protection experiments, showed the photoaffinity labeling to be specific (approximately 1:1) . To identify the residues that were modified on the catalytic subunit, the photoinactivated enzyme was cleaved with CNBr and V8 protease (Staphylococcus aureus) . The resulting peptide fragments were purified by HPLC and were sequenced; these experiments identified the modified residues as Gly-125 and Met-127 . This region of the cAMP-dependent protein kinase catalytic subunit contains many residues that are conserved in serine- and tyrosine-protein kinases.

J Vasc Surg, 1988 Aug, 8(2), 143 - 6
Kinetics of cefuroxime in the groin wound after vascular prosthetic implantation; Sorensen TS et al.; The kinetics of cefuroxime in serum and wound fluid were investigated in the period after vascular prosthetic implantation . Cefuroxime was administered as intravenous bolus injections in doses of 0.75 gm (five patients) or 1.5 gm (five patients) . The concentration of cefuroxime in wound fluid increased after the injection and reached a maximal level corresponding to serum concentration levels with 1.5 hours . The subsequent elimination of cefuroxime from the wound fluid closely paralleled the elimination from the serum . The wound fluid concentrations were found to be greater than the minimum inhibitory concentration for Staphylococcus aureus and Escherichia coli in 8 and 5 hours, respectively, after injection of 0.75 gm of cefuroxime, and in 11 and 7 hours, respectively, after injection of 1.5 gm of cefuroxime.

J Med Microbiol, 1988 Aug, 26(4), 251 - 5
Observations on the resistance to drying of staphylococcal strains; Beard-Pegler MA et al.; Death rates have been determined for staphylococcal strains dried on cotton blanket material and stored at room temperature in the dark and in the light . Methicillin-resistant Staphylococcus aureus (MRSA) strains that produced a golden pigment and had a wide distribution within the hospital survived for longer periods than MRSA strains that produced little pigment and had a restricted local distribution . Death rates of methicillin-sensitive strains of S . aureus at day 7 were similar to those of the general epidemic MRSA strains, and there was no significant difference between the death rates at day 7 of the local epidemic MRSA strains and the coagulase-negative strains.

Infect Immun, 1988 Aug, 56(8), 2193 - 7
Small, antibacterial and large, inactive peptides of neutrophil granules share immunoreactivity to a monoclonal antibody; Marzari R et al.; Monoclonal antibodies were raised against a bactericidal protein fraction that was purified from an extract of bovine neutrophil granules and that was previously shown (A . Savoini, R . Marzari, L . Dolzani, D . Serrano, G . Graziosi, R . Gennaro, and D . Romeo, Antimicrob . Agents Chemother . 26:405-407, 1984) to inhibit bacterial DNA synthesis . One of these antibodies, BP97, was covalently linked to Affi-Gel 10 and was used for immunoaffinity chromatography of granule extracts . Elution of the bound proteins, followed by reversed-phase high-performance liquid chromatography, generated several peptides whose molecular weights fell in the range of 1,600 to 64,000 and which reacted to BP97 but not to other mouse monoclonal or polyclonal antibodies . The reaction to BP97 appeared to be specific, inasmuch as a full panel of cationic oligo- or polypeptides was not recognized by this monoclonal antibody . Among the purified granule polypeptides, the more cationic ones (with molecular weights of 4,300 to 8,000) inhibited the growth of Escherichia coli at a MIC of 12 to 50 micrograms/ml . In addition, a 1,600-molecular-weight, highly cationic peptide also inhibited the growth of Staphylococcus aureus and Staphylococcus epidermidis at MICs of 3 to 8 micrograms/ml.

Int J Food Microbiol, 1988 Aug, 7(1), 25 - 30
The detection of enterotoxin and toxic shock syndrome toxin-1 production by strains of Staphylococcus aureus with commercial RPLA kits; Wieneke AA; Two reversed passive latex agglutination kits (SET-RPLA and TST-RPLA) were used to detect enterotoxin and toxic shock syndrome toxin-1 (TSST-1) production by 334 strains of Staphylococcus aureus isolated from clinical sources and from outbreaks of food poisoning . The results were compared to those obtained with the traditional gel diffusion method . All strains positive by gel diffusion were also positive by RPLA . Seventeen strains produced low levels of TSST-1 or enterotoxins C or D which were only detected by the RPLA method . Some non-specific reactions were obtained with the SET-RPLA kit, but not with the TST-RPLA kit.

Am J Infect Control, 1988 Aug, 16(4), 141 - 6
Attempts to eradicate methicillin-resistant Staphylococcus aureus colonization with the use of trimethoprim-sulfamethoxazole, rifampin, and bacitracin; Roccaforte JS et al.; Retrospective review of 197 patients with methicillin-resistant Staphylococcus aureus (MRSA) identified 47 in whom a regimen for eradication of MRSA colonization could be evaluated . The patients were elderly (mean age, 67.7 years), with 53% transferred from another institution and 53% treated in an intensive care unit . A mean of 47.1 days of hospitalization with an average of 4.9 antibiotics preceded the first MRSA culture . The usual regimen (mean, 6.0 days) was oral trimethoprim-sulfamethoxazole, 160/800 mg twice daily, oral rifampin, 600 mg once daily, and bacitracin ointment three times a day . Eradication succeeded in 40 patients, 9 relapsed, and MRSA persisted in 7 . Twenty-four of 25 nares sites were cleared but only 16 of 22 other sites . MRSA infection eventually developed in 36% . No adverse reactions to the eradication regimen were noted . Although this treatment for MRSA carriage was safe and effective, decreased efficacy outside the nares and relapse limited its value.

Immunology, 1988 Aug, 64(4), 593 - 8
Early lymphocyte activation molecule defined by the monoclonal antibody MLR-3: biochemical and functional studies; Delia D et al.; The MLR-3 monoclonal antibody reacts with activated but not with resting lymphocytes . We report that MLR-3 identifies an early activation molecule since its binding is detectable on T cells 1.5-2 hr after in vitro activation . Its expression, therefore, does not require DNA synthesis and precedes, by many hours, that of the receptors for interleukin-2 (IL-2R) and transferrin (TF-R) . The MLR-3 antigen is also found on activated thymocytes (including the large early thymic CD3- subset) and B cells . The majority of T- and B-lymphoblastoid cell lines, as well as the myeloid and erythroid cell lines HL60, GM1 and K562, are MLR-3+; conversely, non-haemopoietic cell lines are MLR-3 negative . Seventy percent of B-cell chronic lymphocytic leukaemia and 15% of B non-Hodgkin's lymphomas (B-NHL) are MLR-3+ . On tissue sections MLR-3 is reactive with epithelia, sweat glands, hair follicles and Henle's loop but not with vessels, connective, endothelium and many other tissues . In vitro studies show that MLR-3 (1-100 micrograms/ml) significantly alters the thymidine uptake of mitogen-treated lymphocytes:augmentation is found when T and B cells are induced with TPA-Ionomycin and reduction when induced with phytohaemoagglutinin (PHA) or Staphylococcus aureus Cowan strain 1 (SAC), respectively . On SDS-PAGE, MLR-3 immunoprecipitates a disulphide-linked heterodimer of MW 29,000-35,000: both subunits are glycosylated, phosphorylated and exhibit a pI of 4.1 and 5.0, respectively . Our data, particularly the in vitro results, suggest that the MRL-3 molecule could have an important role in the early hours of activation for the progression of resting lymphocytes into mitosis.

Clin Invest Med, 1988 Aug, 11(4), 279 - 85
Effect of HLA-DR positive thyrocytes on in vitro thyroid autoantibody production; Iwatani Y et al.; The function of HLA-DR positive thyrocytes on thyroid autoantibody production has been examined to test the hypothesis that such HLA-DR positive thyrocytes may initiate or aggravate autoimmune thyroid disease . Thyrocytes were cultured (precultured) with leucoagglutinin (which stimulated thyrocyte expression of HLA-DR, beta 2-microglobulin (beta 2-m) and thyroid microsomal antigens) and then cocultured with peripheral blood mononuclear cells . Thyroid antibody production by the latter was then measured . There was no evidence of induction or enhancement of thyroid-microsomal and thyroglobulin autoantibody production in supernatants from the cocultures of autologous peripheral blood mononuclear cells and HLA-DR positive thyrocytes from normal controls and patients with Graves' disease . Furthermore, stimulation of B lymphocytes from patients with autoimmune thyroid disease with a combination of Staphylococcus aureus Cowan I plus supernatants from autologous cocultures of peripheral blood mononuclear cells and HLA-DR positive thyrocytes from normal controls and Graves' disease, produced significantly less microsomal antibody and thyroglobulin antibody than similar cocultures with HLA-DR negative thyrocytes, although total immunoglobulin G (IgG) was similar in both groups . The effect of supernatants from allogeneic cocultures on microsomal antibody thyroglobulin antibody and total IgG production was no different between HLA-DR positive and HLA-DR negative thyrocytes . These data suggest that HLA-DR positive thyrocytes may have a protective role against thyroid autoimmunity rather than a pathogenic role for it.

J Protein Chem, 1988 Aug, 7(4), 377 - 98
Local interactions favor the native 8-residue disulfide loop in the oxidation of a fragment corresponding to the sequence Ser-50-Met-79 derived from bovine pancreatic ribonuclease A; Milburn PJ et al.; A 30-residue peptide was obtained from ribonuclease A by chemical cleavage with cyanogen bromide, subsequent sulfitolysis with concomitant S-sulfonation, and finally enzymatic cleavage with Staphylococcus aureus protease . The peptide was converted to the free thiol form by reductive cleavage of the S-sulfo-protecting groups with D,L-dithiothreitol . This peptide consisted of residues 50-79 of the native sequence of ribonuclease A, with the exception that methionine-79 had been converted to homoserine . Included in this sequence are residues cysteine-65 and cysteine-72, which form a disulfide bond in the native enzyme, as well as cysteine-58 . This molecule may form one of three possible intramolecular disulfide bonds upon thiol oxidation, viz . one loop of 15 and 2 of 8 residues each . These isomeric peptides were prepared by oxidation with cystamine, 2-aminoethanethiolation of residual thiols, and fractionation by reverse-phase high-performance liquid chromatography . Disulfide pairings were established by mapping the tryptic fragments and confirming their composition by amino acid analysis . After protracted incubation under oxidizing conditions at 25.0 degrees C and pH 8.0, the 26-member ring incorporating the native disulfide bond between residues 65 and 72 is the dominant product . Assuming that equilibrium is established, we infer that local interactions in the sequence of ribonuclease A significantly stabilize the native 8-residue disulfide loop with respect to the non-native 8-residue loop (delta G degree = -1.1 +/- 0.1 kcal mole-1) . The implications of this observation for the oxidative folding of the intact protein are discussed.

Postgrad Med J, 1988 Aug, 64(754), 606 - 9
A descriptive survey of uncontrolled methicillin-resistant Staphylococcus aureus in a twin site general hospital; Barrett SP et al.; Over a five year period beginning in 1981, during which control measures were applied intermittently, the incidence of methicillin-resistant Staphylococcus aureus (MRSA) isolates increased steadily within a twin site general hospital . A retrospective chart review of 154 patients identified in 1984-1985 showed that the MRSA 'definitely' contributed to three deaths (2%) and 'probably' contributed to a further 15 (10%) . The prolonged median duration of hospital admission (22 days) before first isolation of MRSA, together with the clustering of cases in time on certain wards, suggested that most, if not all, affected patients acquired the MRSA in hospital . As the virulence of MRSA in our outbreak appeared the same as that reported from teaching hospitals, MRSA control measures need to be comprehensively applied in general hospitals.

J Ultrastruct Mol Struct Res, 1988 Aug, 100(2), 194 - 200
The projection structure of alpha-toxin from Staphylococcus aureus in human platelet membranes as analyzed by electron microscopy and image processing; Olofsson A et al.; Most strains of Staphylococcus aureus produce alpha-toxin, a 33-kDa membrane active protein which is considered to be an important virulence factor of this bacterium . When alpha-toxin interacts with membranes an oligomeric from of the toxin can be seen by electron microscopy as characteristic ring structures in the membrane . A two-dimensional study of these annular structures, incorporated in membranes of human platelets, was performed, introducing a partly new method for rotational alignment of individual particles . It is shown that the averaged oligomer consists of six subunits . At neutral pH the outer diameter of the ring is about 75 A . The stain-filled pore or cavity in the center has a diameter of about 25 A . The size of the hexamer is increased if the pH is lowered.

Biol Chem Hoppe Seyler, 1988 Aug, 369(8), 715 - 25
The amino-acid sequence of rabbit Cu-Zn superoxide dismutase; Reinecke K et al.; The primary structure of Cu-Zn superoxide dismutase from rabbit liver was investigated . The reduced and S-carboxymethylated enzyme was treated with cyanogen bromide, trypsin or Staphylococcus aureus proteinase V8 . The resulting peptides were separated by high-performance liquid chromatography and sequenced by automated Edman degradation . With the exception of the N- and C-terminus the complete sequence was established by means of overlapping peptides . The N-terminus is blocked and thus not susceptible to Edman degradation . The amino-acid composition of the tryptic N-terminal peptide corresponds to that of the cytoplasmatic Cu-Zn superoxide dismutases of other mammals investigated . The chromatographic behaviour of these N-terminal peptides on a reversed phase C18 column is also identical, thus suggesting also for the rabbit Cu-Zn superoxide dismutase the N-terminal sequence Ac-Ala-Thr-Lys . The C-terminus was demonstrated to have the sequence -Ile-Ala-Pro by enzymatic degradation with carboxypeptidase Y . The complete amino-acid sequence of the rabbit Cu-Zn superoxide dismutase consists of 152 amino-acids and shows the expected homology to other Cu-Zn enzymes published so far . The aspartate and six histidine residues known to complex the metal ions are conserved at homologous positions . This also applies for the arginine residue near the C-terminus which is supposed to direct the anionic superoxide radical towards the active centre of the enzyme . The amino acid sequence of the rabbit Cu-Zn superoxide dismutase corresponds to those of other mammals in more than 80% of its amino-acid residues . From a total of 152 amino-acid residues the rabbit shares with rat 128, with mouse 130, with horse 127, with pig 126/127, with cattle 130 and with man 131 amino acids in homologous positions . However the Cu-Zn superoxide dismutases of closely related mammals like rats and mice differ in only five amino acid residues of their sequence . A phylogenetic closer relatedness between lagomorphs and rodents than between other orders of mammals, could not be derived from the sequence data given . Rather rodents and lagomorphs are to be considered as two evolutionary independent orders of mammals.

Antibiot Khimioter, 1988 Aug, 33(8), 587 - 90
{Synergistic action of combinations of penicillin and perhydroacridine derivatives}; Chelysheva GM; The effect of combinations of perhydroacridine derivatives such as 10-nitroso-trans-anti-cys-perhydroacridine (MT-2) and 10-isopropylamino-trans-anti-cys-perhydroacridine (MT-6) with benzylpenicillin on growth of two penicillinase-producing strains of Staphylococcus aureus was studied . The combination components were added to the culture simultaneously or at different periods i.e . the second component was added after preliminary exposure of the bacterial cells to the first component for 1-6 hours at 37 degrees C . It was shown that duration and efficacy of the combination synergistic action were directly proportional to the time of the component addition . The highest synergistic action of the combinations was observed when both the components were simultaneously added to the staphylococcal culture . The combinations were less efficient when the bacterial cells were preliminarily incubated with the perhydroacridines . Addition of the perhydroacridine derivatives after the strain contact with the penicillin resulted in elimination of the combination synergistic action . Thin-layer chromatography did not reveal complexing between the penicillin and perhydroacridines.

Chemioterapia, 1988 Aug, 7(4), 274 - 7
Worldwide assessment of the activity of cefotetan against clinical isolates; Turner PJ et al.; Data on the antibacterial activity of cefotetan collected since 1985 have been compiled into a computer database and an analysis of results on more than 100,000 clinical isolates presented . The studies were conducted in 285 hospitals located throughout Europe, South Africa, Australia and the USA . The results demonstrate the effectiveness of cefotetan as a broad spectrum agent with significant activity against clinically important aerobes and anaerobes . This is exemplified by the susceptibility to cefotetan of 92% of 14,315 strains of Staphylococcus aureus, 99% of 24,103 strains of Escherichia coli and 93% of 1,841 strains of Bacteroides fragilis.

Chemioterapia, 1988 Aug, 7(4), 271 - 3
Cefotetan: antibacterial activity against Staphylococcus aureus in the presence of human serum; Edwards JR; Cefotetan is a broad spectrum cephamycin antibiotic with a long serum half-life (3-4.5 h): this is explained, in part, by serum-protein-binding (SPB) of 88% . The rate of kill of Staphylococcus aureus by cefotetan was assessed in human serum or broth containing reducing concentrations of drug simulating those seen following a 1 g intravenous dose to man . Cefotetan was bactericidal in serum despite the assumed concentration of unbound drug never reaching the minimum inhibitory concentration (MIC) for the test strain . In a separate study, ceftriaxone (SPB 96%) was more active in broth (MIC 4 mg/1) than cefotetan (MIC 16 mg/1) . In 100% serum the minimum bactericidal concentration (MBC) of ceftriaxone was 64 mg/1 whilst the MBC for cefotetan was 16 mg/1.

Am J Vet Res, 1988 Aug, 49(8), 1210 - 3
Sources of variation introduced into a phagocytosis assay as a result of the isolation of neutrophils from bovine blood; Paape MJ et al.; A study was conducted to examine sources of variation introduced into a phagocytosis assay as a result of the isolation of neutrophils from bovine blood, including variation attributable to isolation of neutrophils from blood, variation between duplicate determinations of percentage phagocytosis, and the variation in the ability of neutrophils isolated from blood (over repeated collections from the jugular vein) to phagocytose . For the phagocytosis assay, jugular venous blood from each of 4 cows was divided into 2 equal portions . The neutrophils were isolated by lysis of red blood cells with 0.2% sodium chloride . The neutrophils (2 x 10(7)) were incubated in duplicate with 32P-labeled Staphylococcus aureus ({32P}SA; 2 x 10(8)) in skimmed milk samples (2.5% final concentration) prepared from 4 cows . This process was repeated thrice on neutrophils isolated from 4 cows at 2-week intervals . The proportions of variation in percentage of 32P-labeled S aureus phagocytosed between duplicate neutrophil isolations and between duplicate assay determinations were 0 and 1% . Differences among skimmed milk sources and among runs, using blood neutrophils taken at different times from the same donor cow, accounted for 62 and 36% of the total variation . The results indicated that variation arising from blood neutrophil isolation introduced into a phagocytosis assay within a single-day trial is of no concern . The large variation among skimmed milk sample sources indicated differences among cows in the ability of their milk to support phagocytosis . The variation in neutrophil isolations over time for any cow was considered too large to allow for evaluation of physiologic and environmental effects on phagocytosis of neutrophils isolated from blood.

J Pediatr Surg, 1988 Aug, 23(8), 779 - 81
Osteomyelitis of the cervical spine presenting as a neurenteric cyst; Ein SH et al.; A healthy 3-week-old baby girl developed a cyanotic spell that required intubation and ventilation . During part of her initial emergency examination and treatment, a neck mass was felt, and a positive blood culture grew staphylococcus aureus . She was transferred to the ICU, and was ventilated and treated with intravenous cloxacillin . Bronchoscopy showed a paralyzed left cord . Computerized tomography (CT) scan of her neck showed a midline mediastinal mass (behind the compressed trachea and esophagus), that extended from C7 to the carina . Because of the suspicion of an abnormal C7 vertebral body, diagnosis of a neurenteric cyst was made, and a myelogram showed a complete block at the T1 level and an absent C7 vertebral body . There were no neurologic signs . Her right knee then became red and swollen, and x-rays showed a lytic area in the distal femur . This knee was explored under general anesthesia, and an osteomyelitis found and drained . Several days later, a barium swallow showed the mediastinal mass pushing the esophagus to the left, but several more cervical vertebrae were "missing," and the diagnosis of osteomyelitis of the cervical spine was confirmed . The mediastinal staphylococcal abscess was then drained through the neck . Follow-up has been unremarkable over the last 4 years.

Zh Mikrobiol Epidemiol Immunobiol, 1988 Aug, (8), 79 - 82
{Determination of antibodies to mycobacterial antigens in tuberculosis by erythrocyte immunoadsorption with a rosette marker}; Plaskin DIu et al.; A solid-phase enzyme immunoassay system for the determination of antibodies to mycobacterial antigens, based on the method of erythrocyte immunoadsorption in microchambers for immunological reactions, has been developed . To detect antibodies specifically bound with the solid-phase antigen, the affinity rosettes of Staphylococcus aureus strain Cowan I, carrying protein A, with erythrocytes conjugated with human gamma globulin have been used . The significant correlation of the titers of 34 sera, determined by means of erythrocyte immunoadsorption, with extinction values obtained in the solid-phase enzyme immunoassay of antibodies to Mycobacterium tuberculosis has been established . The coincidence of the results in 92% of cases has been noted.

Eur J Clin Microbiol Infect Dis, 1988 Aug, 7(4), 505 - 10
Interference of Staphylococcus aureus lipase with human granulocyte function; Rollof J et al.; The influence of purified Staphylococcus aureus lipase on granulocyte function and morphology was studied . The lipase itself was strongly chemotactic; in addition preincubation of granulocytes with low concentrations of lipase enhanced the directed movement, as assayed in the agarose system . Higher concentrations of lipase, in contrast, gave a progressive reduction of granulocyte chemotaxis; at 12 micrograms lipase per ml, cells were almost immobilized . Phagocytic killing of Staphylococcus aureus by granulocytes preincubated with lipase was reduced in a dose-dependent manner . At 12 micrograms lipase per ml almost no staphylococcal killing occurred . This was mainly accounted for by a reduction of bacterial uptake, but some decrease in intragranulocytic killing was also noted . These functional alterations, which can all be ascribed to an interference with membrane functions, were associated with marked changes of the granulocyte surface structure, which was denuded and lacked normal microvilli . The effects of lipase were partly retained after heat inactivation of lipase activity, indicating that the effects of staphylococcal lipase on granulocyte function are not due to enzymatic activity alone . These effects of lipase may be an important virulence factor and contribute to the preferential location of lipase-producing Staphylococcus aureus strains at deep sites of infection.

Br J Dermatol, 1988 Aug, 119(2), 189 - 98
Staphylococcal colonization in atopic dermatitis and the effect of topical mupirocin therapy; Lever R et al.; Forty-nine patients with atopic dermatitis entered a double blind placebo controlled cross-over study of mupirocin, a new topical antistaphylococcal antibiotic . Forty-five patients were evaluable . Quantitative bacteriological assessment before treatment showed that heavy colonization of the skin with Staphylococcus aureus was present in nearly all patients even in the absence of overt infection . However, the bacterial count was significantly reduced by 2 weeks' treatment with topical mupirocin, but not by the placebo . Moreover, a significant reduction of clinical severity was also observed after treatment with mupirocin, which was maintained over the following 4 weeks, although recolonization occurred during this period, with bacterial counts rising to pre-treatment levels . Despite recolonization, clinical deterioration was not observed during the trial period . No serious side-effects were observed . Phage typing showed that 50% of patients carried more than one bacterial phage type . Recolonization in eight patients (17%) was with a 'new' strain that had not previously been isolated.

J Immunol, 1988 Aug 1, 141(3), 1034 - 40
Cloning and expression of the H chain V region of antibody OVB3 that reacts with human ovarian cancer; Gallo MG et al.; Hybridoma OVB3 produces an antibody (IgG2b kappa) that reacts with an Ag present on the surface of almost all human ovarian carcinomas . An EcoRI fragment of genomic DNA containing the rearranged H chain V region of monoclonal OVB3 was cloned from a lambda gtWES library and then sub-cloned into a pGEM4 vector . To show that the cloned DNA sequence did encode the V region of a functional antibody, the DNA fragment was inserted into plasmid pSV-VNP gamma SNase in place of VNp to produce pSV-VOVB3 gamma SNase . This plasmid was then transfected into variant OVB3 hybridoma cells, which no longer produced the H chain of antibody OVB3, and functional antibody was secreted by the recipient cells . The recombinant chimeric antibody bound to ovarian cancer cells and contained Staphylococcus aureus nuclease activity, proving that a functional V region gene had been cloned.

J Gen Microbiol, 1988 Aug, 134 ( Pt 8), 2179 - 88
Molecular cloning and genetic analysis of the determinant for gamma-lysin, a two-component toxin of Staphylococcus aureus; Cooney J et al.; The gamma-lysin determinant of Staphylococcus aureus strain Smith 5R has been cloned in phage lambda and plasmid vectors in Escherichia coli . Genetic evidence is presented which demonstrates that gamma-lysin requires the co-operative action of two polypeptides expressed by the closely linked hlgA and hlgB genes . Recombinants expressed haemolytic activity in agarose medium but not in agar, a known property of gamma-lysin . Haemolysis was inhibited by antiserum raised against the 32 kDa component of gamma-lysin, but not by anti-alpha-, anti-beta- or anti-delta-lysin serum . Subcloning and transposon Tn5 mutagenesis identified a 3.5 kb region which was necessary for gamma-lysin expression in E . coli . Two genes (hlgA and hlgB) were mapped and their polypeptide products identified . Non-haemolytic Tn5 mutants fell into two groups based upon complementation tests done between extracts of mutants in vitro and also between extracts of mutants and components of gamma-lysin purified from S . aureus culture supernates . Immunoblotting showed that some mutants in group A (defective in expression of hlgA) did not express a 32 kDa polypeptide which was synthesized by the parental haemolytic recombinant and by mutants in group B . Minicell analysis suggested that the products of the hlgB gene were proteins of 38 kDa and 36 kDa . The smaller molecule co-migrates with a protein in a fraction of the S . aureus culture supernate containing component B of gamma-lysin . The 38 kDa polypeptide is probably an unprocessed precursor . Southern hybridization demonstrated that the hlgA and hlgB genes are closely linked in the chromosome of several strains of S . aureus.

J Appl Bacteriol, 1988 Aug, 65(2), 153 - 61
Comparative studies of selective media for recovery of Staphylococcus aureus from natural waters; Borrego JJ et al.; Several selective media currently used for the enumeration of Staphylococcus aureus from different sources were evaluated in order to establish their quantitative recovery, specificity and degree of selectivity, using different types of water samples . The highest selectivity and reliability in the enumeration of Staph . aureus from the samples was obtained on Borrego-Florido-Romero-0 (BFR-0) and KRANEP agars . The method that produced the highest recovery of Staph . aureus was BFR-0 agar with membrane filter and incubation at 36 degrees C for 48-72 h.

Anaesth Intensive Care, 1988 Aug, 16(3), 338 - 41
Effect of altered environmental temperature on established infection; Lewis GB; An alteration of ten degrees Celsius in environmental temperature significantly alters the mortality from severe established Escherichia coli and Staphylococcus aureus infection in mice . Many patients with severe infection are nursed in wards without air conditioning . It is suggested that even modest levels of environmental stress may influence their recovery.

J Antimicrob Chemother, 1988 Aug, 22(2), 207 - 11
Drug transfer into cerebrospinal fluid after simultaneous administration of ampicillin with ceftriaxone or ceftazidime in rabbits with Staphylococcus aureus meningitis; Okura K et al.; Meningitis was induced in white rabbits with a non beta-lactamase producing strain of Staphylococcus aureus . There was no significant difference in the CSF level of ceftazidime after simultaneous administration of ceftazidime with ampicillin or after ceftazidime alone . In contrast the percentage CSF penetration of ampicillin decreased from 8.81% after administration of ampicillin alone to 2.77% after the simultaneous administration of ampicillin with ceftazidime . The ratio of AUC in CSF and serum was 19.4% after ampicillin alone and 7.71% after simultaneous administration with ceftazidime . When ceftriaxone was combined with ampicillin there was no effect on the CSF penetration of either drug . Cephalosporins have unpredictable effects on the CSF concentration of ampicillin, probably due to variable effects on the penetration of ampicillin from blood to CSF.

J Clin Microbiol, 1988 Aug, 26(8), 1555 - 60
Enzyme-linked immunosorbent assay for Escherichia coli heat-stable enterotoxin type II; Handl C et al.; The gene for Escherichia coli heat-stable enterotoxin type II (STII) was fused to the genes for protein A from Staphylococcus aureus and beta-galactosidase in two different expression systems . Antibodies raised in rabbits against the protein A-STII fusion protein recognized the beta-galactosidase-STII fusion protein . The latter fusion protein was used as the immobilized antigen in an enzyme-linked immunosorbent assay (ELISA) for detection of STII . The correlation between the results of the ELISA and the intestinal loop test in piglets was 95%, suggesting that the ELISA can be used to reliably detect STII.

Circ Shock, 1988 Aug, 25(4), 245 - 58
The effects of chronic bacteremia on the metabolic response to exercise in the conscious rat; Musch TI et al.; Cardiac function was indirectly assessed in a rat model of chronic sepsis by measuring maximal oxygen uptake (VO2max) during exercise . A subcutaneous abscess cavity was created in rats by implanting a gauze sponge in the hindquarter area . Animals with sterile abscesses were compared with rats that had the abscess cavity infected with Escherichia coli and Bacteroides fragilis (gram-negative group) or these two organisms plus Staphylococcus aureus (gram-positive/negative group) . VO2max was measured during exercise before and after the abscess cavity was infected . After a carotid artery cannula was placed, heart rate, mean arterial pressure, arterial blood gases, and acid-base status were measured in three groups of rats (sterile abscess, gram-negative, gram-positive/negative) during submaximal and maximal exercise . Results demonstrated that the infected rats were febrile and had elevated white blood cell counts and intermittent bacteremia, while rats with sterile abscesses did not . VO2max was similar in all groups of rats both before and after the abscess cavity was infected . The metabolic and hemodynamic variables measured during submaximal and maximal exercise in the different groups of rats were similar . These results do not support the contention that gram-positive and/or-negative bacteremia produce cardiac dysfunction during early sepsis.

Immunology, 1988 Aug, 64(4), 709 - 14
Opsonin-dependent and independent surface phagocytosis of S . aureus proceeds independently of complement and complement receptors; Gordon DL et al.; We examined the mechanism of surface phagocytosis of Staphylococcus aureus by human polymorphonuclear leucocytes (PMN) . Surface phagocytosis of unopsonized bacteria occurred, but was significantly enhanced by the presence of serum . The serum requirement was low, and a maximal effect occurred with serum concentrations of 0.25-0.5% . The opsonic effect of serum was not removed by heat inactivation of complement but was adsorbed, at low serum concentrations, by protein A, indicating that opsonin-dependent surface phagocytosis requires IgG but not C3 . The requirement of opsonin-dependent surface phagocytosis for IgG was demonstrated further with purified IgG preparations as the sole opsonin . Activation of PMN by N-formyl-methionyl-leucyl-phenylalanine (FMLP) or phorbol myristate acetate (PMA) increased opsonin-independent surface phagocytosis by 47% and 66%, respectively, but had no effect on opsonin-dependent surface phagocytosis . Blockade of the PMN iC3b receptor (CR3), which has lectin-like properties, by a panel of monoclonal antibodies against the alpha- and beta-chains of CR3 did not inhibit the surface phagocytosis of opsonized or unopsonized S . aureus, and one antibody (NIMP-R10) enhanced opsonin-independent surface phagocytosis . These results indicate that the mechanism of surface phagocytosis is quite different to that observed in suspension assays . Opsonin-independent surface phagocytosis occurs and is enhanced by PMN activation, opsonin-dependent surface phagocytosis is dependent on IgG and not complement, and neither opsonin-independent nor -dependent surface phagocytosis proceeds through CR3.

J Hosp Infect, 1988 Aug, 12(2), 91 - 101
Methicillin-resistant Staphylococcus aureus bacteraemia in Hong Kong; Cheng AF et al.; In the first 22 months of operation at the Prince of Wales Hospital, 26 (46%) of 56 hospital-acquired Staphylococcus aureus bacteraemias were due to methicillin-resistant organisms (MRSA) . There were 10 plasmid profiles amongst 24 of the MRSA strains analysed . MRSA bacteraemias were first seen in the hospital 1 year after opening when the isolation rate of MRSA from all sites had risen to about 1% of patient admissions . During the last 3 months of the study period, 17 out of 18 S . aureus bacteraemias were due to methicillin-resistant strains . Patients with MRSA bacteraemia were significantly more likely than those with methicillin-sensitive S . aureus bacteraemia to have had a severe underlying disease, a poor clinical prognosis, prolonged hospitalization, and prior antimicrobial therapy, especially with aminoglycosides . They also had a significantly longer hospital stay after infection, a significantly higher cost of antimicrobial therapy and a higher mortality rate . The lower mortality rate in MRSA patients treated with vancomycin (18%) compared with those treated with other antimicrobials (40%) confirms that, at present, vancomycin is the treatment of choice for invasive MRSA infections.

Blood, 1988 Aug, 72(2), 672 - 8
B-cell proliferative and differentiative responses after autologous peripheral blood stem cell or bone marrow transplantation; Kiesel S et al.; In this study the authors have evaluated B-cell function after autologous peripheral-blood stem cell transplantation (ABSCT) and autologous bone marrow (ABMT) transplantation . The B-enriched fractions of peripheral blood from ten normal subjects and 22 autografted patients (11 patients after ABMT, eight patients after ABSCT, and three patients after ABSCT followed by ABMT) were investigated . Time postgrafting ranged from 1 to 34 months . Proliferative responses to anti-mu antibody, Staphylococcus aureus Cowan 1 (SAC), and low molecular weight (mol wt) 12-Kd B-cell growth factor (BCGF) were measured . Differentiative responses to the same factors were assessed by quantifying in vitro immunoglobulin (IgG/IgM) production . The authors found no difference in B-cell function between the ABMT and the ABSCT patient groups . Compared to the B cells of normal subjects, only five out of 22 autografted patients showed a normal proliferative response to all agents used, while nine out of 22 did not respond to any signals . Eight out of 22 patients displayed various defects of B-cell response . However, in vitro IgG/IgM secretion of predominantly IgG subclass was normal in 19 out of 22 patients . This in vitro ability to produce Ig was reflected by the patients' normal serum IgG/IgM levels, whereas serum IgA levels were low . The authors speculate that there may be 2 B-cell populations: the normal in vitro Ig production and in vivo serum IgG may come from the stimulation of a small number of re-infused pre-committed memory B cells while, in parallel, immature B cells develop from autografted hematopoietic progenitor cells.

Antimicrob Agents Chemother, 1988 Aug, 32(8), 1192 - 5
DNA gyrase of Staphylococcus aureus and inhibitory effect of quinolones on its activity; Takahata M et al.; DNA gyrase from Staphylococcus aureus FDA 209P was partially purified by lysostaphin treatment, saturation with ammonium sulfate, and affinity chromatography on heparin-Sepharose and with a concentrator (Centricon 30) . It was found to consist of two subunits: alpha and beta . The ability of new quinolone antibacterial agents such as norfloxacin, ofloxacin, and ciprofloxacin to inhibit DNA gyrase activity and cell growth was investigated . The inhibitory effects of the new quinolones against the activity of S . aureus DNA gyrase were in parallel with their antibacterial activities . The 50% inhibitory doses of norfloxacin, ofloxacin, and ciplofloxacin were 0.34, 0.31, and 0.24 micrograms/ml, respectively, while the 50% inhibitory doses of nalidixic acid and cinoxacin, which were less active against S . aureus FDA 209P, were 100 micrograms/ml or more.

Antimicrob Agents Chemother, 1988 Aug, 32(8), 1174 - 81
Genetic analysis of gentamicin resistance in methicillin- and gentamicin-resistant strains of Staphylococcus aureus isolated in Dublin hospitals; Storrs MJ et al.; Methicillin- and gentamicin-resistant strains of Staphylococcus aureus isolated in Dublin hospitals have been classified into groups I, II, and III based on resistance to antimicrobial agents and plasmid profiles . Each group expresses a characteristic level of resistance to gentamicin, tobramycin, and sisomicin . Enzyme assays showed that resistant strains expressed 2"-aminoglycoside phosphotransferase-6'-aminoglycoside transferase activity by a determinant which is known to be chromosomally located . The gentamicin resistance (Gmr) determinants were transferred from group I, II, or III strains by transduction into a laboratory strain where each expressed the same low level of resistance . This finding suggests that high-level resistance in some clinical strains is due to a second, unlinked resistance mechanism . No evidence was obtained by hybridization experiments that clinical isolates or spontaneous mutants expressing high-level Gmr carried more than one copy of the Gmr determinant, thus eliminating the possibility that a gene dosage effect was responsible for high-level resistance . Hybridization experiments with transductants and wild strains suggested that the Gmr determinant was located at homologous sites in wild strains from different groups, although restriction site differences were observed in flanking sequences . Electron microscope analysis of a cloned Gmr determinant and genetic evidence suggested that a Dublin clinical isolate harbored a transposon very similar to Tn4001.

Pediatr Res, 1988 Aug, 24(2), 213 - 6
Phagocytosis-associated functions in neonatal monocyte-derived macrophages; Speer CP et al.; Using a Teflon culture system we analyzed different aspects of cell maturation and phagocytic activities in neonatal monocyte-derived macrophages . Morphological and cytochemical characteristics as well as the protein composition of neonatal macrophages were identical with those of adult controls; actin binding protein (265,000 Da), myosin (210,000), alpha-actinin (102,000) and actin (42,000) could be identified in cells from either source . All phagocytic functions were shown to be perfectly normal in neonatal macrophages when compared with adult cells: random migration, chemotactic response to zymosan-activated serum and formyl-methionyl-leucyl-phenylalanine, ingestion, and killing of Staphylococcus aureus, phagocytosis-associated chemiluminescence, production of oxygen intermediates (superoxide anion, O2-; hydrogen-peroxide, H2O2) . Phorbol myristate acetate-stimulated O2(-)-generation by 1 x 10(5) macrophages was 11.8 +/- 4.7 nmol/h for neonates, and 10.2 +/- 3.9 for controls; production of H2O2 was 7.6 +/- 3.5 nmol/h in neonatal macrophages and 6.4 +/- 2.8 in adult controls . It is unlikely, then, that the increased susceptibility of human neonates to systemic bacterial infections can be related to an abnormality in the essential phagocyte functions of macrophages.

J Clin Microbiol, 1988 Aug, 26(8), 1549 - 54
Particle agglutination assays for rapid detection of fibronectin, fibrinogen, and collagen receptors on Staphylococcus aureus; Naidu AS et al.; Latex beads (0.8-micron diameter; Difco Laboratories) were coated with fibronectin, fibrinogen, collagen type I, or denatured collagen (gelatin) and evaluated in a particle agglutination assay (PAA) for the rapid detection of fibronectin, fibrinogen, or collagen binding to Staphylococcus aureus . These assays were compared with a commercial test for detecting the binding of fibrinogen and immunoglobulin G (Staphaurex) . Bacterial cells (approximately 10(10) cells per ml) suspended in 0.02 M potassium phosphate buffer (pH 6.8) caused the clumping of standard fibronectin, collagen, gelatin, and fibrinogen latex suspensions within 2 min on glass slides . The test results were scored semiquantitatively from strongly positive ( ) to weakly positive (+) and negative (-) reactions . The negative PAA reactions corresponded to a median value of 11.5% relative to the binding of 125I-labeled protein to strain Cowan 1, indicating the high sensitivity of the test . The reactions with fibronectin and fibrinogen latex suspensions and with Staphaurex were optimal for cells grown on tryptic soy and brain heart infusion broth media . Blood agar was optimal for reactions with collagen and gelatin latex suspensions . Media containing high salts (mannitol salt agar and staphylococcus medium 110) enhanced the tendency of cells to autoaggregate . These assays were also clinically evaluated on 187 S . aureus isolates . The PAA reagents were stable, and the assays were highly specific, sensitive, and reproducible, thus making PAA suitable for the rapid screening of the binding of various bacterial pathogens to serum and connective-tissue proteins.

Scand J Immunol, 1988 Aug, 28(2), 177 - 84
Cytomegalovirus-infected adherent cells interact synergistically and antagonistically with Staphylococcus aureus protein A in vitro; Paulin T et al.; Cytomegalovirus (CMV) has been shown to exert suppressive effects on the immune response but also to have mitogenic properties . A bacterial product, protein A from Staphylococcus aureus (SpA) was chosen to study possible interactions in vitro between bacterial products and adherent cells incubated with infectious CMV and ultraviolet light (UV)-inactivated CMV . Small amounts of infectious CMV potentiated SpA-induced DNA synthesis and Ig secretion measured by induction of plaque-forming cells (PFC) . The reason for this may be that CMV in small amounts may act in synergism with the non-specific mitogen SpA . UV-inactivated CMV did not influence these responses except for a markedly enhanced PFC induction with SpA in lymphocytes from seronegative individuals . This remarkable synergism with SpA was also seen in enriched B cells . No synergism was seen in lymphocytes from seropositive donors . Large amounts of infectious CMV markedly reduced SpA-induced immune responses . Preliminary data suggest that the immunosuppressive effects are mediated by an interleukin 1 inhibitory factor . CMV was not shown to be a polyclonal B-cell activator but may, possibly in small amounts, act as such together with bacterial products, which would explain certain autoimmune phenomena . To conclude, CMV could in interaction with a bacterial product generate both synergistic and suppressive effects on immune response.

Biochemistry, 1988 Jul 26, 27(15), 5586 - 92
Evidence for the extramembranous location of the putative amphipathic helix of acetylcholine receptor; Dwyer BP; Evidence has been obtained demonstrating that the peptides GVKYIAE and AIKYIAE found in the potential amphipathic helices of the alpha and beta subunits, respectively, of acetylcholine receptor are not buried in the membrane . The peptide KYIAE was synthesized, and polyclonal antibodies were prepared against a conjugate of bovine serum albumin and synthetic peptide . An immunoadsorbent capable of binding and subsequently releasing peptides ending with the sequence-YIAE was produced by attaching these specific antibodies to agarose . Native acetylcholine receptor was labeled with pyridoxal phosphate and Na{3H}BH4 . The labeled protein was stripped of phospholipid and digested with the protease from Staphylococcus aureus strain V8 . The digest was submitted to immunoadsorption to isolate the labeled indigenous peptides . As a control, alpha and beta polypeptides prepared by gel filtration of a solution of acetylcholine receptor in detergent were stripped of detergent and labeled with pyridoxal phosphate and Na{3H}BH4 in the presence of 8 M urea . The labeled alpha and beta polypeptides were digested and submitted to immunoadsorption . The specific radioactivities of the indigenous peptides from the alpha and beta subunits labeled under native and denaturing conditions were nearly equal . In similar experiments using isethionyl (2', 4'-dinitrophenyl)-3-amino-propionimidate as the labeling agent, the indigenous peptides from native and denatured receptor were also labeled to the same extent . Since these peptides are labeled to the same extent whether or not the protein is denatured, they cannot be buried in the membrane.

J Biol Chem, 1988 Jul 25, 263(21), 10300 - 3
Modulation of the effects of mutations in the basic region of the OmpA signal peptide by the mature portion of the protein; Lehnhardt S et al.; Oligonucleotide-directed site-specific mutagenesis was used to study the structure-function relationship of the positively charged amino terminus of the Escherichia coli outer membrane protein OmpA signal peptide . Mutations were isolated which reduced the overall charge of the amino-terminal region from +2 (wild type) to +1, 0, and -1, as well as one mutation from Thr to Ser at position 4 . DNA encoding the wild type and mutant OmpA signal peptides was then fused in-frame to DNA encoding the mature regions of Staphylococcus aureus nuclease A and TEM beta-lactamase . In the case of both the beta-lactamase and nuclease fusions, normal processing was no longer observed when the charge at the amino terminus was reduced to zero or made negative . Differences between the two hybrid proteins were observed in the case of the Thr to Ser mutation . As expected, this mutation had no effect on the beta-lactamase hybrid; however, the processing rate of the nuclease hybrid protein was reduced to nearly one-half . Furthermore, this effect was essentially reversed when a Lys residue at position 3 was deleted . A model is presented which explains the differing effects of a signal peptide mutation on the secretion of different hybrid proteins based on kinetic differences in the translocation of the nuclease and beta-lactamase proteins.

Am J Med, 1988 Jul 25, 85(1A), 44 - 8
Cefoperazone versus ceftazidime monotherapy of nosocomial pneumonia; Mangi RJ et al.; Cefoperazone and ceftazidime monotherapy were compared in a randomized, prospective evaluation of patients with nosocomial pneumonia . These antibiotics were equally effective, with an overall successful treatment rate of 45 of 62 (73 percent) for cefoperazone-treated patients and 50 of 63 (79 percent) for ceftazidime-treated patients (p = 0.41) . There was no difference in the incidence of side effects (including hypoprothrombinemia), superinfections, or colonization of the oropharynx with yeast, enterococcus, Staphylococcus aureus, or resistant gram-negative bacilli . When antibiotic administration, and laboratory costs are considered, cefoperazone is less expensive than ceftazidime . Both cefoperazone and ceftazidime are effective therapy for nosocomial pneumonia.

Biochim Biophys Acta, 1988 Jul 21, 942(2), 280 - 94
Properties of ion channels formed by Staphylococcus aureus delta-toxin; Mellor IR et al.; The delta-toxin of Staphylococcus aureus has been investigated in terms of its potential to form ion channels in planar lipid bilayers formed at the tip of patch electrodes . Channel formation has been shown to occur for delta-toxin concentrations in the range 0.1 to 2.0 microM . In 0.5 M KCl, two major classes of channels were seen--'small' with conductances of 70-100 pS, and 'large' with a conductance of approx . 450 pS . Current-voltage relationships for lipid bilayers containing several delta-toxin channels revealed both voltage-dependent and independent components to channel gating . Reversal potential measurements showed the channels to be cation selective . In the presence of 3.0 M KCl, the channel gating kinetics were complex, with multiple open and closed states . The results are interpreted in terms of a model for the channel consisting of a hexameric cluster of alpha-helical delta-toxin molecules.

J Biol Chem, 1988 Jul 15, 263(20), 9573 - 5
Structure and bactericidal activity of an antibiotic dodecapeptide purified from bovine neutrophils; Romeo D et al.; Cytoplasmic granules of neutrophils store a variety of cationic polypeptides, which exert in vitro a potent antibacterial action and are potentially involved in host defense mechanisms . From an acid extract of bovine neutrophil granules we have purified over 2,000-fold a dodecapeptide exhibiting bactericidal activity against both Escherichia coli and Staphylococcus aureus at 10(-7)-10(-5) M concentration . The purification procedure involved only two steps of ion-exchange and reversed-phase chromatography . The peptide, named bactenecin, has the amino acid sequence, Arg-Leu-Cys-Arg-Ile-Val-Val-Ile-Arg-Val-Cys-Arg, maintained in a cyclic structure by a disulfide bond between the two cysteine residues . Computer modeling of the dodecapeptide resulted in a conformation in which the chain adopts an antiparallel extended structure forming a gamma turn at residue 7.

J Biol Chem, 1988 Jul 15, 263(20), 9589 - 97
Amino acid sequence of thioredoxin isolated from rabbit bone marrow determined by tandem mass spectrometry; Johnson RS et al.; The amino acid sequence of the thioredoxin isolated from rabbit bone marrow was determined chiefly by high performance tandem mass spectrometry and fast atom bombardment mass spectrometry combined with manual Edman degradation . The sequences of peptides generated by digestion with trypsin alone or in combination with Staphylococcus aureus protease V8 or thermolysin were determined from their collision-induced dissociation mass spectra . Alignment of these sequences and additional sequence information were obtained from the collision-induced dissociation mass spectra of peptides obtained from digestion of the intact protein with S . aureus protease V8 and alpha-chymotrypsin . The resulting sequence of 104 residues is as follows: Val-Lys-Gln-Ile-Glu-Ser-Lys-Ser-Ala-Phe-Gln- Glu-Val-Leu-Asp-Ser-Ala-Gly-Asp-Lys-Leu-Val-Val- Val-Asp-Phe-Ser-Ala-Thr-Trp-Cys-Gly-Pro-Cys-Lys- Met-Ile-Lys-Pro-Phe-Phe-His-Ala-Leu-Ser-Glu-Lys- Phe-Asn-Asn-Val-Val-Phe-Ile-Glu-Val-Asp-Val-Asp- Asp-Cys-Lys-Asp-Ile-Ala-Ala-Glu-Cys-Glu-Val-Lys- Cys-Met-Pro-Thr-Phe-Gln-Phe-Phe-Lys-Lys- Gly-Gln-Lys-Val-Gly-Glu-Phe-Ser-Gly-Ala-Asn-Lys- Glu-Lys-Leu-Glu-Ala-Thr-Ile-Asn-Glu-Leu-Leu.

Biochem J, 1988 Jul 15, 253(2), 489 - 96
Polyclonal and monoclonal antibodies to human lysyl hydroxylase and studies on the molecular heterogeneity of the enzyme; Myllyla R et al.; Human placental lysyl hydroxylase gave two bands in SDS/polyacrylamide-slab-gel electrophoresis: a broad, diffuse, major band corresponding to an apparent Mr of 80,000-85,000, and a sharp minor band with Mr 78,000 . Mouse and chick-embryo lysyl hydroxylases gave only the broad, diffuse band, whereas the sharp band could not be detected . Polyclonal antibodies were prepared to the two bands of the human enzyme separately, and monoclonal antibodies were prepared to the whole purified enzyme preparation . Both types of polyclonal antibody inhibited and precipitated the enzyme activity, and both stained the two polypeptide bands in immunoblotting after SDS/polyacrylamide-gel electrophoresis . Only one out of five monoclonal antibodies inhibited the enzyme activity, whereas they all precipitated the activity when studied with antibody coupled to Sepharose . All five monoclonal antibodies stained the whole broad band in immunoblotting, and at least three of them also stained the sharp band . Peptide maps produced from the two polypeptide species by digestion with Staphylococcus aureus V8 protease were highly similar . Experiments with endoglycosidase H demonstrated that the Mr-80,000-85,000 polypeptide contains asparagine-linked carbohydrate units, which are required for maximal lysyl hydroxylase activity . The data suggest that the lysyl hydroxylase dimer consists of only one type of monomer, the heterogeneity of which is due to differences in glycosylation.

Schweiz Med Wochenschr, 1988 Jul 12, 118(27-28), 1053 - 5
{Fatal peripheral catheter phlebitis}; Widmer A et al.; A 75-year-old man suffered from suppurative thrombophlebitis as a complication of a peripheral venous catheter (1.2 x 45 mm Teflon) . In spite of rapid removal of the catheter at the time of clinical diagnosis of phlebitis and adequate antibiotic treatment, the Staphylococcus aureus sepsis developed into lethal endocarditis . The risk of thrombophlebitis can be minimized by limiting (less than 72 hours) the duration of cannulation . If pus is detected within the lumen of the vein, surgical excision of the involved vein remains the treatment of choice.

Biochemistry, 1988 Jul 12, 27(14), 5227 - 33
Purification of homologous protein carboxyl methyltransferase isozymes from human and bovine erythrocytes; Gilbert JM et al.; We have purified the two major isozymes of the L-isoaspartyl/D-aspartyl protein methyltransferase from both human and bovine erythrocytes . These four enzymes all have polypeptide molecular weights of approximately 26,500 and appear to be monomers in solution . Each of these enzymes cross-reacts with antibodies directed against protein carboxyl methyltransferase I from bovine brain . Their structures also appear to be similar when analyzed by dodecyl sulfate gel electrophoresis for the large fragments produced by digestion with Staphylococcus aureus protease V8 or when analyzed by high-performance liquid chromatography (HPLC) for tryptic peptides . The structural relatedness of these enzymes was confirmed by sequence analysis of a total of 433 residues in 32 tryptic fragments of the human erythrocyte isozymes I and II and of the bovine erythrocyte isozyme II . We found sequence identify or probable identity in 111 out of 112 residues when we compared the human isozymes I and II and identities in 127 out of 134 residues when the human and bovine isozymes II were compared . These results suggest that the erythrocyte isozymes from both organisms may have nearly identical structures and confirm the similarities in the function of these methyltransferases that have been previously demonstrated.

Biochemistry, 1988 Jul 12, 27(14), 5188 - 93
Isolation of multiple forms of DNA polymerase delta: evidence of proteolytic modification during isolation; Lee MY; The subunit structures of a number of human placenta DNA polymerase delta preparations were investigated by Western blotting with polyclonal antisera and by activity staining following polyacrylamide gel electrophoresis . When immunoblots and activity stains were performed on different enzyme preparations, putative catalytic subunits of (a) 170, (b) 120, or (c) 50-70 kilodaltons (kDa) were observed . It was also observed that the lower molecular weight forms could be generated upon storage of the preparations . Western blotting of human placental tissue extracts showed that the major immunoreactive polypeptide was 160-170 kDa . Treatment of the extracts with trypsin or Staphylococcus aureus V8 protease led to the generation of immunoreactive polypeptides of lower molecular weights . These studies suggest that the 120-kDa and lower forms of the enzyme are generated via uncontrolled proteolysis and provide a rationale for the observation of different apparent subunit structures previously reported for DNA polymerase delta . In addition, these findings suggest that DNA polymerase delta has a catalytic domain which resides in a protease-resistant domain.

Biochemistry, 1988 Jul 12, 27(14), 5141 - 9
Mapping of the nucleotide-binding sites in the ADP/ATP carrier of beef heart mitochondria by photolabeling with 2-azido{alpha-32P}adenosine diphosphate; Dalbon P et al.; 2-Azido{alpha-32P}adenosine diphosphate (2-azido{alpha-32P}ADP) has been used to photolabel the ADP/ATP carrier in beef heart mitochondria . In reversible binding assays carried out in the dark, this photoprobe was found to inhibit ADP/ATP transport in beef heart mitochondria and to bind to two types of specific sites of the ADP/ATP carrier characterized by high-affinity binding (Kd = 20 microM) and low-affinity binding (Kd = 400 microM) . In contrast, it was unable to bind to specific carrier sites in inverted submitochondrial particles . Upon photoirradiation of beef heart mitochondria in the presence of 2-azido{alpha-32P}ADP, the ADP/ATP carrier was covalently labeled . After purification, the photolabeled carrier protein was cleaved chemically by acidolysis or cyanogen bromide and enzymatically with the Staphylococcus aureus V8 protease . In the ADP/ATP carrier protein, which is 297 amino acid residues in length, two discrete regions extending from Phe-153 to Met-200 and from Tyr-250 to Met-281 were labeled by 2-azido{alpha-32P}ADP . The peptide fragments corresponding to these regions were sequenced, and the labeled amino acids were identified . As 2-azido-ADP is not transported into mitochondria and competes against transport of externally added ADP, it is concluded that the two regions of the carrier which are photolabeled are facing the cytosol . Whether the two photolabeled regions are located in a single peptide chain of the carrier or in different peptide chains of an oligomeric structure is discussed.

Biochemistry, 1988 Jul 12, 27(14), 5352 - 65
Tubulin structure probed with antibodies to synthetic peptides . Mapping of three major types of limited proteolysis fragments; de la Vina S et al.; We synthesized five peptides homologous to the potentially antigenic positions alpha(214-226), alpha(430-443), alpha(415-443), beta(241-256), and beta(412-431) of the porcine brain tubulin sequences . These peptides were successfully employed to raise tubulin-cross-reactive antibodies . The antibodies are specific of the regions of tubulin spanned by the peptides . They react specifically with the tubulin bands in immunoblots and with microtubules in immunofluorescence assays of cytoskeletons . The peptides of the C-terminal regions have also been employed to localize determinants recognized by two available monoclonal antibodies to tubulin in the positions alpha(415-430) and beta(412-431), respectively . In a first application of the anti-peptide antibodies, we have mapped the fragments of limited proteolysis of purified calf brain tubulin by trypsin, chymotrypsin, papain, thermolysin, subtilisin, and protease V8 from Staphylococcus aureus . Thirty-seven peptides have been identified, of which 32 have been unequivocally aligned into the tubulin sequences on the basis of their antigenic reactivity . There are three major, well-defined zones of preferential cleavage by the proteases: the C-termini and two internal zones in each chain . C-Terminal cleavages of both chains by subtilisin do not remove the antigenic reactivity of the zones alpha(415-430) and beta(412-431) . C-Terminal cleavages by protease V8 are preferential of beta-tubulin . All six proteases tested cleave alpha- and/or beta-tubulin at one or both of the internal zones . These zones are located roughly at one-third and two-thirds of the chain length in both subunits . Therefore, a model of the tubulin monomers is proposed which consists of three major, proteolytically defined, compact regions (N-terminal, middle, and C-terminal thirds) and the cleavable zones . This model is discussed with the tubulin structural information presently available.

J Biol Chem, 1988 Jul 5, 263(19), 9437 - 42
Generation of biologically active interleukin-1 beta by proteolytic cleavage of the inactive precursor; Black RA et al.; Interleukin-1 beta (IL-1 beta) is derived from an inactive precursor by proteolytic cleavage . To study IL-1 beta processing, we expressed the precursor in Escherichia coli, partially purified it, and used it as a substrate for various potentially relevant protease preparations . The precursor alone was virtually inactive, but incubation with membranes from human monocytes or myeloid cell lines yielded a 500-fold increase in IL-1 bioactivity . Western blot analysis of the incubated material showed that the 31,000-Da precursor is broken down to three major products, ranging from 17,400 to about 19,000 Da . The most active of these products is the smallest one, and it co-migrates during electrophoresis with mature IL-1 beta . Four purified known proteases were also tested for their effect on precursor IL-1 beta, and none of these products co-migrated with the mature protein . Chymotrypsin and Staphylococcus aureus protease yielded slightly larger products, which were highly active . Elastase and trypsin yielded substantially larger products, and these had little IL-1 activity . The products of three of the known proteases were identified by NH2-terminal sequencing . These results show conclusively that proteolysis of precursor IL-1 beta generates biological activity and that the cleavage must occur close to the mature NH2 terminus.

J Biol Chem, 1988 Jul 5, 263(19), 9298 - 303
Immunoglobulin binding by the regular surface array of Aeromonas salmonicida; Phipps BM et al.; The cell surface of Aeromonas salmonicida is covered by a regular surface array composed of a single species of protein, the A-protein (Phipps, B . M., Trust, T . J., Ishiguro, E . E., and Kay, W . W . (1983) Biochemistry 22, 2934-2939) . The array, known as the A-layer, is the key virulence factor for this organism . Cells containing the A-layer specifically bound rabbit IgG and human IgM with high affinity (KD = 1.0 X 10(-6) M and 3.3 X 10(-6) M, respectively), but neither isogenic A-protein-deficient strains nor an Aeromonas hydrophila strain also possessing a regular surface array had binding activity . Selective removal of A-protein at pH 2.2 inactivated IgG binding . Structurally intact IgG was requisite for binding since both Fab and Fc fragments were inactive . Aeromonas A-protein did not share the same IgG binding sites as Staphylococcus aureus protein A . Purified A-protein bound IgG only weakly, but reassembled A-layer regained binding activity . Protein modification and perturbation of the A-layer indicated that no single amino acid residue was critical for binding, and that the binding site consisted of a native arrangement of at least four A-protein monomers in the layer.

J Adolesc Health Care, 1988 Jul, 9(4), 325 - 30
Staphylococcus aureus endocarditis in a previously healthy adolescent; Truxal BA et al.; Infectious endocarditis is occasionally a complication of Staphylococcus aureus sepsis in previously well individuals with no heart disease or history of intravenous drug use . We report a case of a 16 year old who developed Staphylococcal sepsis and endocarditis probably as the result of neglected paronychia of her toes . Despite adequate antibiotic therapy, the infectious process destroyed her aortic valve, thereby producing aortic regurgitation complicated by cerebral embolism . Aortic valve replacement surgery was required . Endocarditis should always be sought with S . aureus bacteremia . Intravenous high-dose antibiotic therapy for at least 4 weeks is the recommended therapy.

J Clin Microbiol, 1988 Jul, 26(7), 1425 - 7
Spurious oxacillin resistance in Staphylococcus aureus because of defective oxacillin disks; Boyce JM et al.; Clinical isolates of Staphylococcus aureus wee inappropriately categorized as intermediate or resistant to oxacillin on the basis of tests with two lots of oxacillin disks . The potency of one lot was tested and found to be below accepted limits . Routine quality control tests failed to detect the defective disks.

J Clin Microbiol, 1988 Jul, 26(7), 1331 - 4
Enzyme-linked immunosorbent assay for detection of leukocidin toxin from Staphylococcus aureus in bovine milk samples; Loeffler DA et al.; An enzyme-linked immunosorbent assay was developed for the detection of leukocidin toxin from Staphylococcus aureus . The minimum concentration of leukocidin detectable with the assay was 30 ng/ml . The enzyme-linked immunosorbent assay was found to be a more sensitive method, by a mean of 45-fold, for leukocidin detection than was observation of cytolytic effects of the toxin on bovine neutrophils . A mean toxin concentration of 974 ng/ml was required to produce observable cytolytic effects on neutrophils . Although the enzyme-linked immunosorbent assay was able to detect leukocidin in milk samples from toxin-infused mammary glands, the toxin was detectable in only 2 of 27 S . aureus-infected milk samples (7%) from cows with chronic staphylococcal mastitis . To determine whether leukocidin antibodies in the mastitic milk samples were preventing toxin detection, leukocidin was mixed with milk with a high antileukocidin antibody titer (from a vaccinated cow) and evaluated with the immunoassay . Leukocidin was readily detected in this sample, indicating that milk antileukocidin antibodies were not sufficient to prevent detection of any leukocidin present in the mastitic milk samples . Failure to detect leukocidin in most mastitic milk samples with this assay indicated that, if leukocidin is produced in the bovine mammary gland during chronic staphylococcal mastitis, the concentration of the toxin may be too low to produce cytolytic effects on neutrophils.

J Clin Microbiol, 1988 Jul, 26(7), 1257 - 62
Prospective study of microbial colonization of the nose and skin and infection of the vascular access site in hemodialysis patients; Kaplowitz LG et al.; We conducted a prospective study of nasal and skin floras in 71 patients receiving chronic hemodialysis . We wished to determine whether a sterile skin preparation technique was more effective than a clean technique in removing microorganisms from the skin of the vascular access site . We also examined the effect of administration of antibiotics and status of patient hygiene on microbial flora . The presence of Staphylococcus aureus in the nose had a low predictive value for the simultaneous presence of the microorganism on the skin . The status of skin colonization can be accurately assessed only by culture of the skin . Sterile technique was no more effective at removing microorganisms from skin than was clean technique . Antibiotics significantly affected nasal flora but not skin flora . S . aureus was significantly more likely to remain on the skin after application of an antiseptic in patients with poor hygiene than in patients with good hygiene (P = 0.002) . Patients with poor hygiene also had a significantly higher concentration of S . aureus on the skin of the vascular access site after application of antiseptic than patients with good hygiene (P = 0.005) . We found no evidence to support a change from clean to sterile technique for skin preparation, but improvement in personal hygiene may be an effective strategy for prevention of vascular access infections.

Cornell Vet, 1988 Jul, 78(3), 281 - 300
Studies on gangrenous mastitis in goats; Abu-Samra MT et al.; One hundred and fifty goats were diagnosed as having proven gangrenous mastitis . The disease was categorized into early, intermediate and late stages . Gangrenous mastitis in goats is typified by a sudden onset, dark hyperemia, and edema with progressive discoloration of the distal part of the udder . The disease affected lactating goats but not the dry ones . Coagulase positive Staphylococcus aureus was isolated from 60% of composite and half milk samples obtained from the diseased goats . The histopathological changes mainly comprised proliferation of connective tissue, thrombosis and necrosis involving a group of lobules . Treatment of the early and intermediate stages of the disease was successful through the administration of systemic and intramammary terramycin together with diuretics and topical antiseptic cream . The late stage of the disease was successfully treated only through surgery.

Cornell Vet, 1988 Jul, 78(3), 243 - 52
The effects of corticosteroid administration on the migration, phagocytosis and bactericidal capacity of equine neutrophils; Morris DD et al.; Neutrophil function was evaluated in six clinically normal adult horses, immediately before and 3-6 hours after they were given one dose of hydrocortisone sodium succinate (1 mg/kg body weight) . Random migration, stimulated migration to zymosan-activated serum, bacterial phagocytosis and bactericidal capacity of neutrophils were determined in vitro . The mean indices of stimulated migration (net migration and migration ratio) were significantly greater after CS administration (net migration = 62 +/- 23 micron; migration ratio = 11.5 +/- 6.7) than before CS administration (net migration = 44 +/- 10 micron; migration ratio = 6.0 +/- 3.1; P less than 0.05) . Random migration, bacterial phagocytosis and bactericidal capacity of neutrophils were unchanged by CS therapy . Results from this study suggest that the migration of equine neutrophils is influenced, but not impaired, after one dose (1 mg/kg) of hydrocortisone sodium succinate and that the latter causes no change in the ability of equine neutrophils to phagocytize and kill Staphylococcus aureus.

Am J Trop Med Hyg, 1988 Jul, 39(1), 103 - 4
Milkborne gastroenteritis due to Staphylococcus aureus enterotoxin B from a goat with mastitis; Gross EM et al.; Three children developed Staphylococcus aureus food poisoning due to enterotoxin type B following ingestion of milk from a goat with overt mastitis.

Med Decis Making, 1988 Jul-Sep, 8(3), 165 - 74
Is there a role for surgery in the acute management of infective endocarditis? A decision analysis and medical claims database approach; Abrams HB et al.; In the absence of good clinical evidence from a randomized trial, the authors performed a decision analysis to determine the potential value of early elective surgery (OPNOW) for patients with left-sided Staphylococcus aureus infective endocarditis . Initial impressions (before performance of decision analysis) and initial runs at the formal models using probability estimates derived from clinicians suggested that OPNOW (i.e., within a few days of starting antibiotics) offered no advantage over attempted medical cure (WAIT) (life expectancy: WAIT = 325 weeks; OPNOW = 255 weeks) . Extensive sensitivity analyses identified critical variables that needed further empirical estimation . The Manitoba Health Services Commission database identified 127 incident cases of endocarditis between April 1, 1979, and March 31, 1985, enabling estimation of values for these critical variables . With these estimates, the early surgery strategy appeared much better than the previous analyses had suggested (life expectancy: WAIT = 208 weeks, OPNOW = 256 weeks) . The authors believe that this approach of combining decision analysis with medical claims databases is useful as an alternative or precursor to randomized trials, especially where the resource requirements and logistic difficulties of performing randomized trials are great.

Eur J Biochem, 1988 Jul 1, 174(4), 663 - 70
New insights into maitotoxin action; Sladeczek F et al.; Maitotoxin (3 ng/mol) induced a massive uptake of 45Ca2+ into BC3H1 cells . This effect exhibits a lag phase of 3 min . Inositol diphosphate formation occurred concomittantly with the 45Ca2+ uptake but inositol monophosphate formation was found only after a 5-min delay following toxin addition . Maitotoxin-induced 45Ca2+ influxes could not be blocked by either 1 microM verapamil, 1 microM nifedipine or 1 mM La3+ but was blocked by Zn2+ (IC50 = 41 microM) . In addition to inositol phosphate formation and 45Ca2+ uptake, maitotoxin stimulated a large uptake of Na+ and a great loss of K+ in BC3H1 cells . In the absence of Ca2+ (1 mM EGTA) none of the four maitotoxin effects could be detected . After restoration of Ca2+, the maitotoxin effects reappeared even when the toxin itself was no longer present . The divalent cation, Co2+ (1 mM), inhibited ion movements induced by maitotoxin and also digitonin (8.1 microM) . The toxin action showed a very pronounced pH dependence . At low pH, maitotoxin was inactive . The dose-response curves for H+ ion inhibition of maitotoxin-induced Ca2+ uptake showed a shift to the right when determined in the absence of HCO3- and HCO3-/Cl- ions . It was concluded that the primary action of maitotoxin in BC3H1 cells was a pore-forming or channel-forming activity of a non-classical type . Some properties of maitotoxin resemble those of alpha-latrotoxin, others those of pore-forming agents such as melittin or alpha-toxin of Staphylococcus aureus.

J Bacteriol, 1988 Jul, 170(7), 2954 - 60
Cloning and nucleotide sequence of the type E staphylococcal enterotoxin gene; Couch JL et al.; The gene for staphylococcal enterotoxin type E (entE) was cloned from Staphylococcus aureus into plasmid vector pBR322 and introduced into Escherichia coli . A staphylococcal enterotoxin type E-producing E . coli strain was isolated . The complete nucleotide sequence of the cloned structural entE gene and the N-terminal amino acid sequence of mature staphylococcal enterotoxin type E were determined . The entE gene contained 771 base pairs that encoded a protein with a molecular weight of 29,358 which was apparently processed to a mature extracellular form with a molecular weight of 26,425 . DNA sequence comparisons indicated that staphylococcal enterotoxins type E and A are closely related . There was 84% nucleotide sequence homology between entE and the gene for staphylococcal enterotoxin type A; these genes encoded protein products that had 214 (83%) homologous amino acid residues (mature forms had 188 {82%} homologous amino acid residues).

Infect Immun, 1988 Jul, 56(7), 1785 - 91
Alteration of sleep in rabbits by Staphylococcus aureus infection; Toth LA et al.; Abundant evidence suggests that sleep might be altered during infectious disease, although the relationship between sleep and infectious disease has never been examined systematically . To address this issue, we determined the effects of Staphylococcus aureus infection on rabbit sleep . Rabbits inoculated intravenously with S . aureus demonstrated the expected physiological changes consistent with a state of infectious disease (e.g., lymphopenia, neutrophilia, and fever), as well as time-dependent changes in sleep patterns . The sleep changes were characterized initially by increases in (i) the time spent in slow-wave sleep, (ii) the electroencephalographic slow-wave amplitudes during slow-wave sleep, and (iii) the duration of individual bouts of slow-wave sleep . At 20 to 36 h after inoculation, sleep responses fell to levels below corresponding control values for 6 to 12 h . At 6 to 10 h after inoculation, rapid-eye-movement sleep was suppressed and remained at low levels throughout the remainder of the 48-h recording period . These effects of bacterial infection on sleep were attenuated by antibiotic (cephalothin) therapy . Inoculation with killed bacteria produced similar changes in sleep and other physiological parameters, although significantly higher numbers of organisms were required to produce equivalent responses . We postulate that changes in sleep may represent an adaptive response of the host to infectious disease.

Arch Intern Med, 1988 Jul, 148(7), 1608 - 10
Pyomyositis in a patient with the acquired immunodeficiency syndrome; Gaut P et al.; Pyomyositis is an acute bacterial infection of skeletal muscle . It is a common disease in the tropics; fewer than 50 cases of pyomyositis have been reported in the continental United States . Most patients are healthy males, although the disease has been reported in diabetics and in the immunocompromised . This article presents the first detailed known reported case of pyomyositis in a patient with the acquired immunodeficiency syndrome; Staphylococcus aureus was the etiologic agent.

J Med Microbiol, 1988 Jul, 26(3), 189 - 97
Evaluation of electrophoretic methods for typing methicillin-resistant Staphylococcus aureus; Gaston MA et al.; Three electrophoretic methods of typing methicillin-resistant Staphylococcus aureus (MRSA) strains--plasmid profiles (PP), whole-cell protein profiles (WCPP) and immunoblotting profiles (IP)--were evaluated and compared with phage typing . The results obtained with isolates from 12 outbreaks were compared both within the outbreaks, to determine the consistency of results, and between outbreaks . There was generally good agreement between the typing methods but in only six outbreaks did all four methods indicate the same relationship between isolates . WCPP comprised more than 50 bands; when differences occurred, they were seen in only a few bands . In contrast, IP comprised only one or two major bands and the differences were much easier to interpret . The PPs of many of the isolates were similar; many isolates contained a plasmid of mol . wt (18-25) x 10(6) . In several outbreaks both WCPP and IP showed minor differences between isolates that were not apparent with phage typing . When comparisons were made between the 12 index strains and an isolate representing the London epidemic MRSA strain, phage typing and WCPP were the most discriminatory methods; both gave nine distinct patterns, whereas there were eight IPs and only six PPs amongst the 13 strains . It was concluded that both WCPP and IP could provide valuable epidemiological data on MRSA and that IP was the easiest of the three methods to interpret.

J Lab Clin Med, 1988 Jul, 112(1), 16 - 22
Specific binding of Staphylococcus aureus to cultured porcine cardiac valvular endothelial cells; Johnson CM et al.; Infective endocarditis usually occurs in patients who have had previous cardiac damage or who have congenitally abnormal hearts . However, this infection may afflict otherwise normal individuals, and it is often caused by Staphylococcus aureus . In these individuals, interactions between circulating microorganisms and resident cardiac endothelial cells may initiate the infection . In the present studies we established an assay to measure in vitro binding of S . aureus to porcine cardiac valve endothelial cells . We found that this interaction was specific and saturable with respect to time . In contrast, there was no specific binding of Escherichia coli, an organism that rarely causes endocarditis . Exogenous fibronectin had no effect on specific binding of S . aureus, and heat-killed organisms adhered equally well as viable bacteria . Fixation of the endothelial cells with formalin abolished all specific binding . Soluble components from bacterial extracts inhibited S . aureus binding in dose-dependent fashion . These observations suggest that circulating S . aureus may interact with specific sites on cardiac endothelial cells, thereby potentially initiating infective endocarditis.

Ann Emerg Med, 1988 Jul, 17(7), 736 - 8
Toxic shock syndrome following diagnostic peritoneal lavage; Catapano M et al.; We report the case of a 15-year-old girl who developed high fever, syncope, abdominal pain, nausea and vomiting, myalgia, pharyngitis, and a desquamating rash eight days after a diagnostic peritoneal lavage . The diagnostic peritoneal lavage wound was erythematous and tender . Incision of the site yielded 10 mL of exudate that cultured Staphylococcus aureus . The patient was treated with a first-generation cephalosporin and recovered without sequelae . To our knowledge, this is the first reported case of toxic shock syndrome following diagnostic peritoneal lavage.

Reg Immunol, 1988 Jul-Aug, 1(1), 56 - 61
Recombinant bovine interferon-alpha I 1 inhibits the migration of lymphocytes from lymph nodes but not into lymph nodes; Kalaaji AN et al.; Recombinant bovine interferon-alpha I 1 (IFN-alpha) was tested for effects on lymphocyte migration into and out of the lymph nodes of sheep . Chronic lymph drainage methods were used to monitor the efferent lymph of single lymph nodes of unanaesthetized animals . When IFN-alpha was injected into the drainage area of the lymph node, infused directly via an afferent lymphatic, or injected intravenously, a marked decrease in cell output was observed . Nanogram quantities of IFN-alpha produced an effect that lasted several hours . To test whether this effect was specific for lymphocytes, we first stimulated a lymph node with Staphylococcus aureus so that large numbers of neutrophils and lymphocytes were present in efferent lymph . When IFN-alpha was then injected, the lymphocytes disappeared from efferent lymph but the neutrophils did not . Normal efferent lymph cells were labeled with radioisotopes and injected intravenously during the IFN-alpha induced effect . The entry of lymphocytes from the blood was significantly enhanced at a time coinciding with the suppressed cell exit in the efferent lymph . Recovery data indicated that the lymphocytes were retained temporarily in the lymph node but were not destroyed . Furthermore, the bovine IFN-alpha does not appear to be conspicuously antigenic in sheep . It is not yet certain whether IFN-alpha enhances the interactions of cells within the lymph node thereby potentiating immunologic reactions.

J Clin Immunol, 1988 Jul, 8(4), 296 - 306
Intestinal nodular lymphoid hyperplasia in patients with common variable immunodeficiency: local accumulation of B and CD8(+) lymphocytes; Van den Brande P et al.; Common variable immunodeficiency (CVI) with hypogammaglobulinemia is often complicated by nodular lymphoid hyperplasia of the intestine . In this study the lymphoid constituents of intestinal nodular hyperplasia of five CVI patients were characterized with monoclonal antibodies . Few CD4(+) but abundant CD8(+) T lymphocytes were found around the follicles . The follicles were populated mainly by B cells expressing surface IgM . A few cells in the lamina propria expressed Leu7 . No intracytoplasmic immunoglobulin-containing plasma cells were seen . Peyer's patches in gut biopsies from controls were also composed of follicles with B lymphocytes . A ring of T lymphocytes surrounded the follicles . CD4(+) helper cells largely outnumbered CD8(+) cells in this ring . Moreover, plasma cells were present in the lamina propria and the mixed cell zone covering the follicles . In peripheral blood of the patients, B cells were present in normal proportions but they could not be induced to produce IgG in vitro by T cell-dependent (pokeweed mitogen) or T cell-independent (Staphylococcus aureus Cowan I) mitogens . In two of the patients, IgM production could be induced in vitro . Peripheral blood T cells were predominantly CD8(+) in three of the five patients, and in these same patients an increase in suppressor-cell activity of peripheral blood T cells on immunoglobulin production was observed . The data demonstrate a block in B-cell differentiation in the gut and in peripheral blood . Whether the local increase in CD8(+) cells in the nodular lymphoid hyperplasia is a primary event or is secondary to chronic immune stimulation and whether it contributes to local inhibition of B-cell differentiation remain to be investigated.

Blood, 1988 Jul, 72(1), 116 - 20
Further specificity characterization of von Willebrand factor inhibitors developed in two patients with severe von Willebrand disease; Lopez-Fernandez MF et al.; Circulating inhibitors against von Willebrand factor (vWF) that show the properties of heterologous IgG antibodies have been described in a few patients with severe von Willebrand disease (vWD) . The present study provides further characterization of inhibitors from two patients with severe vWD . Inhibitors in both, like polyclonal rabbit antibody, detected all sizes of multimers and the complex structure of each multimer from platelets and plasma of normal individuals as well as from plasma of patients with IIA, IIB, and IIC vWD . Both inhibitors and the rabbit antibody reacted mainly with the intact 225-Kd vWF subunit and the 189-H and 140-Kd fragments in contrast to monoclonal antibodies specific for vWF fragments that detected a higher relative proportion of 176-Kd fragment . Furthermore, all these antibodies recognized fragment III, although one inhibitor and rabbit polyclonal antibody reacted poorly and the other inhibitor did not react at all with reduced fragment II of vWF digested with Staphylococcus aureus V-8 protease . These data suggest that although human inhibitors from severe vWD patients may behave, to some extent, as polyclonal heterologous antibodies against native vWF, the former show striking differences in their target specificity as well as a much broader specificity than that described for human factor VIII inhibitors.

Vet Surg, 1988 Jul-Aug, 17(4), 182 - 5
Chlorhexidine diacetate and povidone-iodine cytotoxicity to canine embryonic fibroblasts and Staphylococcus aureus; Sanchez IR et al.; Chlorhexidine diacetate and povidone-iodine were evaluated for fibroblast toxicity on a primary line of canine embryonic fibroblasts, and for bactericidal efficacy against Staphylococcus aureus . The cultured fibroblasts or S . aureus were exposed for 30 minutes to incremental dilutions of 0.5 and 0.0005% chlorhexidine diacetate, 5.0 to 0.05% povidone-iodine, or physiologic buffered saline as a control . To determine survival, fibroblasts were trypsinized and counted; S . aureus colonies were counted on brain-heart infusion agar . Survival for both groups was expressed by calculating the number of living cells in test dilutions as a percentage of the number in control cultures . Fibroblast survival occurred at chlorhexidine concentrations less than 0.013% and at povidone-iodine concentrations less than 0.5% (p less than 0.05) . Significant S . aureus survival (p less than 0.05) was noted at chlorhexidine concentrations less than 0.05% and povidone-iodine concentrations less than 1.0% . These data showed that all bactericidal concentrations of chlorhexidine diacetate and povidone-iodine were lethal to canine embryonic fibroblasts in vitro, whereas non-lethal concentrations allowed significant bacterial survival.

Rev Argent Microbiol, 1988 Jul-Sep, 20(3), 125 - 35
{Inhibitory effect of Staphylococcus aureus on the growth of rat sarcoma}; Dlugovitzky DG et al.; We have studied the effect upon the growth of a transplantable rat sarcoma, of an intraperitoneal inoculation of peritoneal exudate cell (PEC), spleen cells (SC) and non-adherent spleen cells (N-ASC) obtained from S . aureus previously inoculated rats and the same cells from normal rats (NPEC, NSC, NN-ASC) . Inbred, adult rats were inoculated with 800 x 10(6) bacteria, killed by tyndalization . After 7 days, treated and normal animals were sacrificed and PEC, SC and N-ASC were obtained . Different groups of animals were inoculated with these cell populations . Simultaneously the rats received a s.c . inoculum of a transplantable sarcoma (S-E 100) . Tumor size was measured on days 4, 7, 14, 21 and 28 after tumor challenge . A significant tumor growth inhibition was found with all three cell populations, but no effect was observed with normal cells . Tumor size on different days and tumor growth curves clearly demonstrate this effect . We conclude that Staphylococcus aureus inoculum as well as the cells from animals previously challenged with the bacteria induce tumor growth inhibition . The possibility that protein A of S . aureus may be involved in this phenomenon, or that the mechanism of tumor growth inhibition is mediated by an activation of different cell populations is discussed.

Diagn Microbiol Infect Dis, 1988 Jul, 10(3), 145 - 8
In vitro susceptibility to 10 antibiotics of methicillin-resistant (MRSA) Staphylococcus aureus in Chile; Montiel F et al.; The in vitro activity of 10 antibiotics was determined for 231 strains of methicillin resistant (MRSA) Staphylococcus aureus . Oxacillin was a very good antibiotic to determine methicillin-resistance . Its agreement with methicillin-resistance was in all the strains tested . On the contrary, the correlation with nafcillin was established only in 95% of the strains tested . Cloxacillin and flucloxacillin are not good methicillin-resistance indicators . The strains tested against macrolides, such as erythromycin, and lincosamides, such as lincomycin and clindamycin presented a susceptibility of 68, 78, and 80%, respectively . All tested strains were susceptible to vancomycin.

Res Vet Sci, 1988 Jul, 45(1), 16 - 21
Vaccination against experimental staphylococcal mastitis in ewes; Watson DL; Experiments were carried out in ewes using a new vaccine developed for the prevention of mastitis caused by Staphylococcus aureus . The vaccine comprised three major components: (i) killed S aureus cells which had been cultured to induce synthesis of pseudocapsule; (ii) toxoided staphylococcal beta haemolysin and (iii) the adjuvant dextran sulphate . Ewes systemically vaccinated twice during pregnancy developed significantly elevated circulating levels of IgG1 and IgG2 anti-pseudocapsule antibody, as well as increased serum titres of anti-beta haemolysin . Five different strains of S aureus were used to challenge both vaccinated and control ewes by the intramammary route during the ensuing lactation . The incidence of acute gangrenous mastitis and nonacute, clinical mastitis was significantly lower in vaccinated than in control groups after challenge with each strain . Vaccinated ewes produced significantly more milk than control ewes after challenge with four of the five strains of S aureus.

Rev Infect Dis, 1988 Jul-Aug, 10 Suppl 2, S351 - 5
Surface proteins of Staphylococcus aureus; Cheung AL et al.; Crude cell walls of Staphylococcus aureus of potential immunologic or epidemiologic importance were prepared for study by mechanically disrupting whole cells and applying differential centrifugation . Cell-wall proteins were then released from the wall by lysostaphin digestion and analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis . Distinct electrophoretic protein patterns were found for each of the 12 serologically distinct strains examined . There were two major groups of cell-wall proteins identified in the 45-57-kilodalton (kdal) and 100-190-kdal range, respectively . Proteins in the 45-57-kdal range appeared to represent protein A variants of different apparent molecular mass . Localization experiments with 125I labeling demonstrated the surface location of most of the proteins . These data suggest that selected surface-exposed proteins may be important in defining host immune responses to S . aureus infections.

Med Trop (Mars), 1988 Jul-Sep, 48(3), 243 - 7
{Antibiotic sensitivity of strains of Staphylococcus aureus isolated in Lome}; De Souza C et al.; Use of antibiotics in infectious diseases is frequent . But such a use is not always based on antibiograms and is often not justified . One of the serious consequences of it against public health is the selection of multiresistant strains . So, the authors carried out some researches to determine the profiles of resistance against antibiotics of the strains of S . aureus which is a pathogen frequent in Togo . Sensitivity of the strains was studied in vitro by diffusion method with gelose . The results show that out of 29 antibiotics tested, the 7 most efficient are: rifampicin (91.5 p.c.), sisomicin (91 p.c.), gentamicin (85.64 p.c.), dibekacin (85.8 p.c.), rifamycin (85.71 p.c.) . On the contrary, these strains are resistant against tetracycline (96.02 p.c.), colistin (93.39 p.c.) and doxycycline (92.39 p.c.) . consequently, selective pressure of these three antibiotics is very high at Lome . Restriction or temporary suppression of their use commands attention in view of preventing the spread of such resistances.

Bioorg Khim, 1988 Jul, 14(7), 893 - 904
{Ribonuclease Fl1 from Fusarium lateriticum . Isolation, substrate specificity and amino acid sequence}; Bezborodova SI et al.; Extracellular RNase Fl1 has been purified from the culture filtrate of Fusarium lateritium . The enzyme has been obtained in the electrophoretically homogeneous state with the yield about 90% and 300 fdd degree of purification . RNase Fl1 is a guanyl specific enzyme (EC 3.1.27.3) with the specific activity on RNA 1420 units/mg of protein . The total primary structure of the RNase has been determined by the automated Edman degradation of two non-fractionated peptide hydrolysates produced by trypsin and Staphylococcus aureus protease and of the hydroxylamine cleavage products of the protein . It was shown that hydroxylamine converts the RNase Fl1 N-terminal residue, pyroglutamic acid, into the hydroxyamic acid derivative sensitive to Edman degradation . RNase Fl1 consists of 105 amino acid residues (Mr 10,852) and is a structural homologue of the Fus . moniliforme RNase F1, differing from the latter by 15 amino acid substitutions outside the enzyme active site.

J Clin Microbiol, 1988 Jul, 26(7), 1283 - 6
Staphylococcal scalded skin syndrome in two immunocompetent adults caused by exfoliatin B-producing Staphylococcus aureus; Opal SM et al.; An exfoliatin B-producing strain of Staphylococcus aureus was isolated from two adults with typical staphylococcal scalded skin syndrome (SSSS) . One patient developed desquamation after a local staphylococcal infection of the hand, and the other developed exfoliation after nosocomially acquired staphylococcal endocarditis . Neither patient was immunocompromised, had evidence of renal insufficiency, or manifested other potential risk factors for SSSS . Purified toxin, isolated from the causative organisms, produced a Nikolsky sign in neonatal mice . The toxins were shown to be exfoliatin B by biochemical and immunologic methods and heretofore had been described only in children with SSSS . Analysis of plasmid DNAs from both strains revealed a 23-megadalton plasmid with identical restriction endonuclease digestion fragments . One isolate belonged to phage group II (3B/3C/6/7/47/54/55), whereas the other isolate belonged to phage groups I and III (7/29/52/52A/53/54/80) . The observation that a non-phage group II exfoliatin-producing strain of S . aureus may produce SSSS in adults indicates the need to better define the diagnostic criteria for SSSS . Immunocompetent adults may remain susceptible to some strains of exfoliatin B-producing S . aureus.

J Surg Res, 1988 Jul, 45(1), 37 - 43
Interferon-gamma restores immune competence after hemorrhagic shock; Livingston DH et al.; Hemorrhagic shock increases the susceptibility to infection in both clinical and laboratory settings . Hemorrhagic shock also is associated with a decreased production of interferon-gamma (IFN-gamma), a potent modulator of immune function . We investigated the effect of IFN-gamma both alone and in addition to antibiotic prophylaxis upon infection following hemorrhagic shock . Sprague-Dawley rats were bled to a mean arterial pressure of 45 mm Hg for 45 min and then were resuscitated with shed blood and normal saline . Abscess formation was induced 1 hr later by subcutaneous injection of 1 X 10(8) Staphylococcus aureus . Four treatments were investigated: (1) control; (2) recombinant rat IFN-gamma, 7500 units, 30 min after inoculation and daily for 3 days; (3) cefamandole (CEF) nafate, 30 mg/kg, 30 min before and 4 hr after inoculation; and (4) IFN-gamma + CEF as in (2) and (3) . Abscess size, weight, and quantitative bacterial counts were measured 7 days after inoculation . Hemorrhagic shock increased mean abscess size from 11.7 +/- 2.8 to 14.1 +/- 1.9 mm (P less than 0.05), in untreated rats . IFN-gamma alone resulted in minor changes in abscess formation in both shocked and unshocked animals . Shock rendered CEF ineffective in reducing abscess size . IFN-gamma + CEF significantly reduced abscess size (14.1 +/- 1.9 to 8.1 +/- 1.8 mm) and weight (771 +/- 214 to 252 +/- 132 mg) and decreased bacterial count after shock to 12% of control (all P less than 0.05) . These data demonstrate that hemorrhagic shock impairs antibiotic efficacy; however, the addition of IFN-gamma restores the ability of host defenses to combat bacterial infection.

Rev Med Univ Navarra, 1988 Jul-Sep, 32(3), 163 - 8
{Protein A: an immunomodulator in cancer patients}; Subira ML et al.; Protein A defined as one of the components of the wall of Staphylococcus Aureus Cowans 1 has shown its effects as modifier of the immune response: it induce changes in cellular receptors, polyclonal activation of T and B lymphocytes, liberation of lymphokines, increase in the production of gamma-interferon (probably as the result of activation of NK cells) . In the present work we have studied NK activity and its modifications by Protein A and gamma-interferon in blood of 50 patients with solid tumours . The method used was the Cr 51 liberation assay . The effectors cells were non-adherent lymphocytes treated with different doses and times of incubation of Protein A and gamma-interferon . As target cells we utilized the cellular line K-562 for NK activity and autologous tumoral cells isolated from surgical specimen for tumour-specific cytotoxicity . The results obtained allow us to state the following conclusions: 1 . NK activity against K-562 cells is not always a specific parameter of such activity . 2 . Tumour-specific cytotoxicity can vary according to the histological type of tumour . 3 . Protein A is a potent inducer of tumour-specific cytotoxicity in a higher degree than gamma-interferon.

J Antimicrob Chemother, 1988 Jul, 22 Suppl A, 45 - 62
Clinical application of the newer beta-lactam antibiotics; Turck M; A variety of new beta-lactam antibiotics is in use today . A considerable number of new penicillins have extended spectra which include Pseudomonas and Klebsiella spp . and other important nosocomial organisms . In addition, clavulanic acid, a beta-lactamase inhibitor, has been added to some of these compounds to enhance further their spectrum against Staphylococcus aureus and other bacteria . The first-, second- and third-generation cephalosporins are described . Second-generation cephalosporins have an extended spectrum, covering Gram-negative organisms . Third-generation compounds have both improved beta-lactamase stability and better Gram-negative cover . Other types of beta-lactam antibiotics have recently been developed, such as imipenem and aztreonam, which also have broad activity against Gram-negative bacteria.

Antimicrob Agents Chemother, 1988 Jul, 32(7), 1012 - 8
Enhanced phagocytosis, killing, and serum sensitivity of Escherichia coli and Staphylococcus aureus treated with sub-MICs of imipenem; Adinolfi LE et al.; The influence of pretreatment of Escherichia coli and Staphylococcus aureus with sub-MICs of the new beta-lactam antibiotic imipenem on phagocytosis and killing by murine peritoneal macrophages and the susceptibility of these organisms to serum bactericidal activity were studied . The effects of imipenem, a round form inducer in gram-negative rods, and piperacillin, a filamentous form inducer, were compared . Bacteria grown in the presence of sub-MICs of imipenem or piperacillin were incubated for 30 min with macrophage monolayers in the absence of antibiotic . Phagocytosis, killing, and survival within macrophages were evaluated by microbiological and fluorescence microscope assays . Bacteria grown in the presence of a sub-MIC of imipenem were phagocytized and killed in numbers significantly higher than untreated or piperacillin-treated bacteria were . Intracellular bacteria pretreated with a sub-MIC of imipenem were also readily killed by lymphokine-activated macrophages . Prior treatment with a sub-MIC of imipenem resulted in an increased susceptibility of E . coli but not S . aureus to the bactericidal activity of immune serum . Imipenem treatment and immune serum acted synergistically to enhance phagocytosis and killing . The data indicate that exposure of E . coli and S . aureus to a sub-MIC of imipenem enhances the susceptibility of these potential pathogens to cellular and humoral host defense mechanisms.

Clin Exp Immunol, 1988 Jul, 73(1), 155 - 9
The effect of random blood transfusions on immunoglobulin production by peripheral blood mononuclear cells from uraemic patients; Degiannis D et al.; The effect of blood transfusion on humoral immunity in chronic renal failure was studied by examining immunoglobulin production in vitro, in patients awaiting renal transplantation . Pokeweed mitogen (PWM) induced IgG plaque formation was normal in non-transfused uraemic patients while both spontaneous and Staphylococcus aureus Cowan I (SAC) induced immunoglobulin production were reduced . Five to ten units of third party blood transfusion reduced PWM-driven B cell differentiation, but had no effect on SAC-induced plaque formation, while spontaneous production of immunoglobulin was either enhanced or unaffected . As it is known that the response to SAC is less affected by suppressor T cell activity than that to PWM, these differences in the inhibitory effects of blood transfusion on B cell differentiation are further evidence that transfusion may act by increasing suppressor T cell activity.

J Laryngol Otol, 1988 Jul, 102(7), 656 - 7
Bacterial tracheitis in a young adult; Ruddy J; A previously healthy young adult presented with inspiratory stridor and hoarseness but minimal dysphagia . Indirect laryngoscopy and lateral neck X-rays confirmed a diagnosis of membranous tracheitis . This responded to humidification, antibiotics and steroids . Secretions removed at direct laryngoscopy sent for culture grew Staphylococcus aureus . The literature is reviewed.

Oral Surg Oral Med Oral Pathol, 1988 Jul, 66(1), 124 - 9
Total joint replacement: a consideration for antimicrobial prophylaxis; Cioffi GA et al.; Infection is the principal and most devastating complication of total joint replacement, resulting in long periods of hospitalization, staggering costs, loss of the implant, disastrous physical impairment, and even death . Staphylococcus aureus and Staphylococcus epidermidis account for more than 50% of late infections . Animal studies have shown that joint implants are at a high risk of becoming infected via a metastatic hematogenous route during transient bacteremias . Because cephalosporins have been established as the perioperative and intraoperative agents of choice to prevent infections related to total joint replacement, oral cephalosporins are the drugs of choice to minimize the potential for the metastatic infection of prosthetic joints associated with transient dental bacteremias . Clindamycin is preferred for patients who are allergic to the cephalosporins or who may have a cross-allergy between penicillin and the cephalosporins.

Surgery, 1988 Jul, 104(1), 41 - 8
Opioids modulate human neutrophil and lymphocyte function: thermal injury alters plasma beta-endorphin levels; Deitch EA et al.; To investigate the role of opioids in the acquired immune dysfunctional state that occurs after burns or trauma, plasma beta-endorphin levels were measured serially in nine severely burned patients, and the effect of four different opioids on normal neutrophil and lymphocyte function was quantitated . The rationale for these studies is that the neuroendocrine system appears capable of interacting with and modulating immune function . The plasma levels of beta-endorphin increased to higher than normal during the first 36 hours after burn (15 versus 3.4 pmol/L, p less than 0.05) but quickly returned toward normal . Morphine had the most profound effect on in vitro neutrophil function; it decreased neutrophil chemotaxis but increased neutrophil bactericidal activity for Staphylococcus aureus, as well as resting and zymosan-stimulated oxygen consumption . Other opioids (naloxone, met-enkephalin, and beta-endorphin) had no direct effect on neutrophil chemotaxis or bactericidal activity . Both naloxone and met-enkephalin increased neutrophil oxygen consumption in a dose-dependent fashion, whereas beta-endorphin impaired neutrophil oxygen consumption . None of the opioids altered resting lymphocyte blastogenesis . The only opioid that impaired the ability of normal lymphocytes to respond to mitogen stimulation at physiologically relevant doses was beta-endorphin . These results, documenting that beta-endorphin levels are altered after thermal injury and that opioids can modulate normal neutrophil and lymphocyte function in vitro, support the concept that changes in neuroendocrine activity may occur and potentially alter immune function.

J Vasc Surg, 1988 Jul, 8(1), 1 - 9
Experimental colonization of a polyester vascular graft with Staphylococcus aureus: a quantitative and morphologic study; Leport C et al.; Colonization of a polyester (Dacron) vascular graft by Staphylococcus aureus 209P-R was studied . Twenty-five dogs had thoracoabdominal aortic bypass . After intervals of 2 hours (three dogs), 8 days (five dogs), 1 month (six dogs), 2 months (six dogs), or 6 months (five dogs), a bacteremic challenge was produced by intravenous injection of 6 x 10(8) colony-forming units of S . aureus . Two hours later grafts were removed and cut into 10 fragments, each submitted to bacterial counts and scanning electron microscopic studies . Results of bacterial counts were expressed in colony-forming units (CFU) per square centimeter of graft segment (median {lower to upper quartiles}) . Normal canine aortas (n = 2) used as controls trapped no bacteria . Colonization of Dacron grafts varied according to the duration of graft function (p less than 0.01): after 2 hours, 4416.5 CFU (1158 to 9073 CFU); after 8 days, 1515 CFU (963 to 2893 CFU); after 1 month, 199 CFU (86 to 538 CFU); after 2 months, 615 CFU (243 to 1407 CFU); and after 6 months, 1 CFU (1 to 5 CFU) . Heavily colonized fragments were observed for duration of graft function of 2 months or less, whereas at 6 months all the fragments trapped fewer than 50 CFU/cm2 of graft segment . Scanning electron microscopy showed that colonization was closely associated with healing . Staphylococcal entrapment was related to the amount of fibrin deposits, which were especially abundant where the thrombotic matrix was unorganized and on bare polyester filaments . Graft colonization is especially to be feared in the first weeks after graft implantation, an observation which may help to define guidelines for preventing hematogenous vascular graft infection.

J Hosp Infect, 1988 Jul, 12(1), 29 - 34
Cross-infection between animals and man: possible feline transmission of Staphylococcus aureus infection in humans?
Scott GM, Thomson R, Malone-Lee J, Ridgway GL.
An outbreak of epidemic methicillin-resistant Staphylococcus aureus occurred on a rehabilitation geriatric ward . Intensive screening of patients and staff revealed an unusually high carriage rate in the nursing staff (38%), thought to be related to a ward cat which was heavily colonized from the environment . Infection control measures and removal of the cat led to rapid resolution of the outbreak.

J Pharm Pharmacol, 1988 Jul, 40(7), 509 - 11
Sustained anti-adherence activity of taurolidine (Taurolin) and noxythiolin (Noxyflex S) solutions; Blenkharn JI; Taurolidine (2% w/v) and noxythiolin (1% w/v and 2.5% w/v) solutions inhibit the adherence in-vitro of Escherichia coli and Staphylococcus aureus to human epithelial and fibroblast cells . This effect, demonstrable after 30 min exposure of cells to test drugs, persists after removal of the active compound . Significantly reduced adherence of bacteria is apparent for 5 h after taurolidine treatment and for 6 h after treatment with 2.5% noxythiolin . Anti-adherence activity of taurolidine and noxythiolin may contribute to the observed clinical efficacy of these agents.

APMIS, 1988 Jul, 96(7), 596 - 600
Lipoxygenase products in experimental septic arthritis in dogs; Herlin T et al.; Induction of an experimental septic arthritis was performed in the juvenile dog knee by an intra-articular injection with Staphylococcus aureus . From plasma, lipoxygenase (LO) products of arachidonic acid were separated by reversed-phase high-performance liquid chromatography (RP-HPLC) after extraction of lipids . The plasma samples did not contain UV-detectable amounts of LO products other than the LTB4 metabolite 20-COOH-LTB4, and 12-HETE . Small amounts of chemokinetically active material were found coeluting with the eluate fraction of 20-COOH-LTB4, LTB4 and 12-HETE after RP-HPLC . After 48 hours of septic arthritis, resulting in a massive acute joint inflammation, no significant changes in LO products were observed.

Proc Natl Acad Sci U S A, 1988 Jul, 85(13), 4874 - 8
Recombinant human interferon-gamma reconstitutes defective phagocyte function in patients with chronic granulomatous disease of childhood; Sechler JM et al.; Monocytes from 19 of 30 patients with the classic phenotype of chronic granulomatous disease of childhood (CGD) responded to 3 days of treatment in culture with recombinant human interferon-gamma (rHuIFN-gamma) at 100 units/ml by producing superoxide after stimulation with phorbol 12-myristate 13-acetate . Cells from 15 of 16 patients with cytochrome b-positive CGD (15 with autosomal and 1 with X chromosome-linked inheritance) and cells from 4 of 14 patients with cytochrome b-negative CGD (13 with X chromosome-linked and 1 with autosomal recessive inheritance) responded . Subcutaneous rHuIFN-gamma (0.01-0.05 mg/m2) administered as a single dose, daily or every other day, for five or six doses to 3 patients whose phagocytes responded to rHuIFN-gamma in vitro resulted in significant improvement in phagocyte bactericidal activity against Staphylococcus aureus and increases in superoxide production . Studies on 1 patient's cells indicated the increases in superoxide production correlated with increased membrane cytochrome b . The effects of rHuIFN-gamma persisted for more than a week following cessation of therapy . Thus, we have demonstrated a partial correction in vivo of these CGD patients' phagocyte defect with rHuIFN-gamma . Moreover, the data suggest that a significant proportion of patients with CGD will respond to rHuIFN-gamma with augmentation of phagocyte microbicidal function.

J Immunol, 1988 Jul 1, 141(1), 164 - 73
Inhibitory influence of IL-4 on human B cell responsiveness; Jelinek DF et al.; The role of IL-4 in human B cell activation, proliferation, and differentiation was examined . rIL-2, but not rIL-4, was able to promote maximum proliferation and generation of Ig-secreting cells in cultures of highly purified B cells stimulated with Cowan I Staphylococcus aureus (SA) . Addition of rIL-4 to rIL-2-supported cultures of SA-stimulated peripheral blood, spleen, or lymph node B cells dramatically suppressed both proliferation and differentiation . Results from experiments in which rIL-4 was added to culture at progressively later times indicated a requirement for rIL-4 to be present during the first 2 days of a 5-day incubation to cause inhibition of responsiveness . When a two-stage culture system was utilized, rIL-4 was found to support proliferation or differentiation of B cells initially activated with SA for 2 days only minimally . However, rIL-4 did not inhibit responses of SA preactivated B cells supported by IL-2 . The presence of rIL-4 during the initial 48-h activation of B cells with SA and rIL-2 resulted in a profound inhibition of the ability of the activated B cells to respond subsequently to rIL-2 or lymphokine-rich T cell supernatants . A similar 48-h incubation with rIL-4 alone without SA had no effect on subsequent B cell responsiveness . The presence of rIFN-gamma during B cell activation decreased the inhibitory effect of IL-4 . Other cytokines including IFN-alpha, IL-1, and commercially available low m.w . B cell growth factor also diminished the inhibitory effect of IL-4 . These results indicate that IL-4 inhibits the capacity of human B cells to be activated maximally by SA and rIL-2 and therefore suggest a new immunomodulatory role for this cytokine.

J Neurosurg, 1988 Jul, 69(1), 110 - 4
The pathogenesis of spinal epidural abscess: microangiographic studies in an experimental model; Feldenzer JA et al.; An experimental model of spinal epidural abscess was developed in rabbits by injecting Staphylococcus aureus into the posterior thoracolumbar epidural space . This model has been shown to reproduce the neurological, bacteriological, and radiological aspects of the human disease . In this study, the effect of the infectious epidural mass on the vasculature of the spinal cord in paraplegic rabbits was studied using microangiographic techniques . The normal vascular anatomy of the rabbit spinal cord was defined in control experiments . Vascular proliferation was demonstrated in the epidural space surrounding the abscesses . Anterior and paired posterior spinal arteries remained patent in paraplegic rabbits with mild or moderate spinal cord compression and in some cases of severe compression . In animals with severe compression, the anterior epidural venous plexus remained patent, but the dorsal spinal vein was occluded . Occlusion of perforating arteries occurred only with extreme spinal cord compression . These data indicate that the initial neurological deficit associated with experimental spinal epidural abscess is not due to vascular thrombosis.

Gene, 1988 Jun 30, 66(2), 183 - 92
High-level expression of a functional human fibrinogen gamma chain in Escherichia coli; Bolyard MG et al.; Human fibrinogen gamma chain has been expressed intact at high levels in Escherichia coli . The construction of the expression plasmid, p253, is described . Synthesis of the recombinant protein is isopropyl-beta-D-thiogalactopyranoside-dependent and is driven by the tac promoter . Western analysis of E . coli lysates demonstrates a novel protein of approx . 45 kDa which cross-reacts with antisera to human fibrinogen gamma chains . The protein is not soluble in common E . coli lysis buffers and becomes soluble in 6 M guanidine.HCl or 6 M urea . Initial insolubility is due to interchain disulfide bond formation and to noncovalent interactions . Induced cells examined by phase-contrast microscopy contain dense inclusion bodies . A known function of the gamma chains of human fibrinogen is the clumping of Staphylococcus aureus Newman D2C cells {Hawiger et al., Biochemistry (1982) 1407-1413} . We demonstrate that suspensions of recombinant gamma chains retain the ability to clump cells from this strain of S . aureus.

Biochemistry, 1988 Jun 28, 27(13), 4753 - 60
Amino acid sequences of substrate-binding sites in chicken liver fatty acid synthase; Chang SI et al.; The amino acid sequences of three essential regions of chicken liver fatty acid synthase have been determined: that around 4'-phosphopantetheine ("carrier" site), the substrate "loading" site containing serine, and a "waiting" site for the growing fatty acid containing cysteine . The amino acid sequence of the 4'-phosphopantetheine region was determined for the acetyl-, malonyl-, hydroxybutyryl-, and butyryl-enzyme with peptides obtained by hydrolysis of the enzyme with trypsin and Staphylococcus aureus (V8) protease . The sequence region around the essential serine was obtained for the acetyl- and malonyl-enzyme . The N-terminus of the tryptic peptide was blocked . However, the same sequence is obtained for the acetyl- and malonyl-peptide after S . aureus protease digestion, suggesting that the enzyme contains a single acyl transferase rather than two separate transacylases . The sequence around the cysteine was obtained by use of a radioactive iodoacetamide label . An unusual sequence of three serines adjacent to the cysteine was found . The strong similarities between peptides from different species for all three of the regions suggest that the multifunctional polypeptides from yeast and animals have evolved from the monofunctional enzymes of lower species.

J Biol Chem, 1988 Jun 25, 263(18), 9050 - 4
Immunological characterization of thioltransferase from pig liver; Gan ZR et al.; Polyclonal antibodies against pig liver thioltransferase were raised in a New Zealand rabbit . These antibodies completely neutralized the thioltransferase activity of the homogeneous enzyme and that in the crude cytosolic homogenate at an equivalent titer . The antibodies also cross-reacted equally with calf thymus glutaredoxin and calf liver thioltransferase, but not with Escherichia coli thioredoxin, suggesting that thioltransferase and glutaredoxin from the same species are identical . Immunoblotting analysis of the cytosolic proteins from 14 different pig tissues revealed that most pig tissues contain a 12-kDa protein which reacts with these antibodies . This protein is found in greater abundance in stomach, small intestine, liver, skeletal muscle, kidney, heart, lung, and cerebral cortex, whereas retina, cerebellum, spleen, pancreas, and thymus have low levels of the protein . No reactive protein was detected in the lens . The tissue distribution of the protein was also determined by assay of the enzyme activity and was generally in good agreement with that obtained from the immunoblotting survey . Pig liver thioltransferase was cleaved by trypsin, chymotrypsin, Staphylococcus aureus V8 protease, and cyanogen bromide . The selected peptides purified by reversed phase high performance liquid chromatography or ion exchange fast protein liquid chromatography were subjected to reaction with the polyclonal antibodies against pig liver thioltransferase . Four antigenically reactive fragments were detected by dot-blotting analysis . These peptides are located in the first 30-amino acid residues from the NH2 terminus and the sequence from amino acid residues 39-67, indicating that the active site of the enzyme, Cys22 and Cys25, is located on one of the antigenic determinant domains.

J Am Vet Med Assoc, 1988 Jun 15, 192(12), 1721 - 5
Efficacy of clindamycin in the treatment of Staphylococcus aureus osteomyelitis in dogs; Braden TD et al.; The efficacy of clindamycin in the treatment of experimentally induced, posttraumatic Staphylococcus aureus osteomyelitis was studied in dogs . At the end of the experiment, bacteria could not be isolated from bone marrow of 15 of 16 (93.7%) dogs treated with clindamycin, whereas bacteria could not be isolated from similar specimens obtained from 6 of 13 (46.1%) untreated dogs . None of the 16 dogs treated with clindamycin had histopathologic evidence of osteomyelitis at the end of the experiment . Five of the 13 untreated control dogs had histopathologic evidence of osteomyelitis . The recovery rate was 31% in untreated dogs, whereas 94% of dogs treated with clindamycin recovered from osteomyelitis . Clindamycin, 11 mg/kg of body weight, given orally, q 12 h, for 28 days, was efficacious in the treatment of experimentally induced, posttraumatic S aureus osteomyelitis in dogs.

Biochem J, 1988 Jun 15, 252(3), 683 - 91
Effect of specific proteolytic cleavages on tubulin polymer formation; Serrano L et al.; The capacity for self-polymerization and shape of the tubulin polymers assembled after digestion with trypsin, Pronase, chymotrypsin, subtilisin, Staphylococcus aureus proteinase V8 and proteinase K were investigated . Digestion with trypsin, Pronase or chymotrypsin resulted in a decrease in the ability of tubulin for self-assembly, whereas limited proteolysis with subtilisin, S . aureus proteinase V8 or proteinase K resulted in an increase in such ability . The shape of the assembled polymers varied from typical microtubules (after the treatment with trypsin or Pronase) to sheets (after the treatment with chymotrypsin) and from hooked microtubules with a constant polarity (after the treatment with subtilisin) to the disappearance of a defined polarity of such polymers (after the treatment with S . aureus V8 proteinase or proteinase K) . These results indicate that the tubulin C-terminal regions are involved in the regulation of microtubule polymerization, shape, directional growth and lateral interactions between tubulin protofilaments.

Antibiot Khimioter, 1988 Jun, 33(6), 440 - 3
{Antibiotic AL-87 induction of changes in the fatty acid composition of phospholipids and neutral lipids in a sensitive strain of Staphylococcus aureus 209P}; Smirnov VV et al.; Under the action of the sub-bacteriostatic concentration (0.02 microgram/ml) of antibiotic AL-87 there formed in all the fractions of phospholipids and neutral lipids of S . aureus 209P unsaturated branched fatty acids not detected in the control and the content of shorter chain saturated branched fatty acids increased . This means that there were changes leading to increased lipid fluidity . The findings showed that fatty acids of the neutral lipids and phospholipids were involved in regulation of the bacterial cell functional state and participated in this case in providing increased membrane fluidity and permeability.

Antimicrob Agents Chemother, 1988 Jun, 32(6), 932 - 5
Antiseptic and antibiotic resistance plasmid in Staphylococcus aureus that possesses ability to confer chlorhexidine and acrinol resistance; Yamamoto T et al.; Plasmid pSAJ1 from a methicillin- and gentamicin-resistant strain of Staphylococcus aureus had am molecular size of 50 kilobases and conferred resistance not only to kanamycin, gentamicin, tobramycin, amikacin, benzalkonium chloride, acriflavin, and ethidium bromide but also to chlorhexidine . In addition, the cloned antiseptic resistance gene(s) manifested acrinol resistance in Escherichia coli.

J Antimicrob Chemother, 1988 Jun, 21(6), 773 - 86
Evaluation of antibiotic effectiveness against Staphylococcus aureus surviving within the bovine mammary gland macrophage; Sanchez MS et al.; Bovine mastitis due to Staphylococcus aureus may become chronic and refractory to antibiotic therapy because of the organism's ability to survive within the mammary gland macrophages and/or polymorphonuclear neutrophils (PMNs) . Therefore, phagocytosis and killing of S . aureus by bovine udder macrophages, udder and blood neutrophils and blood monocytes were studied . Gland and blood PMNs were about equally effective at phagocytosing (2.5 log reduction in supernatant) and killing the bacteria (92% reduction of viable bacteria by two hours) . Gland macrophages phagocytosed at a lower rate (1.5 log reduction) and were less effective at killing the bacteria (73% reduction by two hours) . Blood monocytes phagocytosed and killed S . aureus at the lowest rate . An udder macrophage monolayer system was developed and used to evaluate the ability of antibiotics to kill surviving intracellular S . aureus . This assay was similar to our previously described system with blood PMNs . Several classes of antibiotics were investigated . These included naphthalenic ansamycin, lincosaminide, tetracycline, coumarin, peptide, paulomycin, quinolone, macrolide, cephalosporin, and penicillin-class antibiotics . The activity of these compounds was compared to positive (rifampicin), negative (cloxacillin), and nonantibiotic treated controls . Only naphthalenic ansamycin class antibiotics, paulomycin, paldimycin and ciprofloxacin caused significant reduction in viable intracellular bacteria in the macrophage system . These results were similar to those obtained in the blood PMN monolayer system . Because a low intraphagolysosomal pH could affect an antibiotic's ability to kill intracellular bacteria by affecting the drug itself or inhibiting bacterial growth, the effect of low pH on the minimum inhibitory concentration and the minimum lethal concentration of clindamycin and rifampicin against three strains of S . aureus was also tested . While the activity of clindamycin at pH 5.0 compared to pH 7.0 was not affected greatly, the activity of rifampicin was greatly enhanced at acidic pH . These results suggest that at least some of the excellent activity of rifampicin for intracellular S . aureus is due to potentiation of its activity in the intracellular acidic compartment of the phagolysosome.

Chem Phys Lipids, 1988 Jun, 47(2), 117 - 22
Substrate specificity of Staphylococcus aureus (TEN5) lipases with isomeric oleoyl-sn-glycerol ethers as substrates; Kotting J et al.; For the first time fully protected substrates with only one hydrolyzable ester bond have been used to analyze the substrate specificity of microbial lipases . In these substrates the ester is attached to the glycerol molecule in a precisely defined position . The use of three different substituents generates chirality and thus allows the analysis of positional specificities of individual lipases . Therefore, these new substrates have been used to study the enzymatic activities of two closely related lipases isolated from Staphylococcus aureus (TEN5) designated the 44 and 43 kDa lipase . The lipases, especially the 44 kDa molecule, show a high specificity for the hydrolysis of the ester in the sn-1 position (S-configuration), which is hydrolyzed by a factor of ten faster than that in the sn-3 position . In addition, the study demonstrates for the first time that the rate of hydrolysis of a fatty acid ester attached to the sn-2 position of glycerol by microbial lipases depends on the configuration of the substrate molecule.

Am J Vet Res, 1988 Jun, 49(6), 851 - 5
In vitro depression of bovine lymphocyte function by treatment of cultured bovine lymphocytes with physiologic concentrations of hydrocortisone; Ojo-Amaize EA et al.; The effect of hydrocortisone (hydrocortisone sodium succinate) on bovine lymphocyte blastogenesis in response to Staphylococcus aureus antigens and phytohemagglutinin was measured in vitro . Lymphocytes isolated from the blood of cows were treated for 6 to 8 days with physiologic hydrocortisone concentrations known to be inducible by environmental stress (10 ng/ml), acute clinical mastitis (25 ng/ml), or adrenocorticotropin treatment (45 ng/ml) . All 3 concentrations of hydrocortisone caused a depression (P less than 0.01) in lymphocyte blastogenesis in response to phytohemagglutinin and S aureus antigen extract . Hydrocortisone concentrations as low as 10 pg/ml caused a depression in the lymphocyte blastogenic response to phytohemagglutinin . Marked variation existed among cows in the normal response of their nontreated lymphocytes and in the degree of depression of lymphocyte function after the in vitro treatment with hydrocortisone . Macrophage depletion experiments showed that the suppressive effect of hydrocortisone was not mediated by induction of suppressor macrophages . The data suggest that T-cell function was impaired directly by hydrocortisone treatment.

J Antimicrob Chemother, 1988 Jun, 21 Suppl D, 101 - 6
Efficacy and tolerability of erythromycin acistrate and erythromycin stearate in acute skin infections of patients with atopic eczema; Salo OP et al.; The efficacy and tolerability of a new erythromycin derivative, erythromycin acistrate (EA), were compared with that of erythromycin stearate (ES) in 42 patients with infected atopic eczema . The dosage of EA was 400 mg tid and that of ES 500 mg tid . The duration of treatment ranged from five to 12 days . The patients were hospitalized and evaluated before treatment and on the last day in hospital . The infective pathogen was usually Staphylococcus aureus in both groups . Without local antibacterial treatment both drugs eradicated the bacteria in more than 60% of the cases . Gastrointestinal side effects were frequently reported with both drugs, more often in the ES- than in the EA-group, but the difference was only statistically significant (p less than 0.05) with respect to diarrhoea . One patient in each group discontinued treatment because of gastrointestinal side effects . No elevations in liver enzymes of clinical significance were reported in either group.

J Clin Microbiol, 1988 Jun, 26(6), 1236 - 7
Chromatofocusing in the purification of staphylococcal enterotoxin D; Lei Z et al.; A chromatofocusing procedure for the purification of staphylococcal enterotoxin D was developed . The purification included the removal of the toxic protein from culture supernatant fluids of Staphylococcus aureus 1151m by batch adsorption with CG-50 resin, chromatofocusing on Polybuffer Exchanger 94, and gel permeation chromatography on Sephacryl S-200 . The purity of the staphylococcal enterotoxin D obtained was approximately 98%.

J Clin Microbiol, 1988 Jun, 26(6), 1223 - 4
Rapid detection (4 h) of methicillin-resistant Staphylococcus aureus by a bioluminescence method; Park CH et al.; A 4-h bioluminescence method for methicillin susceptibility determination was compared with reference methods . Of the Staphylococcus aureus strains tested, 80 were methicillin resistant, 180 were methicillin susceptible, and 10 were borderline susceptible . There was 100% correlation between bioluminescence and reference methods for methicillin-susceptible and methicillin-resistant strains . All borderline-susceptible strains were identified as methicillin resistant by bioluminescence.

J Steroid Biochem, 1988 Jun, 29(6), 559 - 69
Comparative analysis of estrogen receptors covalently labeled with an estrogen and an antiestrogen in several estrogen target cells as studied by limited proteolysis; Elliston JF et al.; Estrogen receptors covalently labeled with the estrogen affinity label {3H}ketononestrol aziridine (KNA) or with the antiestrogen affinity label {3H}tamoxifen aziridine (TAZ) were subjected to limited proteolysis with trypsin, alpha-chymotrypsin, and Staphylococcus aureus V8 protease and then analyzed on 10-20% sodium dodecyl sulfate-polyacrylamide gradient gels followed by fluorography . The similar molecular weights of intact receptors (Mr 66,000 daltons) and the proteolytic digest patterns indicate extensive homology among estrogen receptors from MCF-7 human breast cancer cells, GH4 rat pituitary cells and rat uterus when liganded with estrogen or antiestrogen . Each protease generated a distinctive ladder of estrogen receptor fragments, and the fragmentation patterns were virtually identical for estrogen receptors labeled with estrogen (KNA) or antiestrogen (TAZ) . Each protease yielded a relatively "resistant" receptor fragment of about 28,000-35,000 daltons . Trypsin and chymotrypsin at higher concentrations generated a much smaller 6,000-8,000 dalton digest product that still contained the {3H}KNA- or {3H}TAZ-labeled receptor binding site . Moreover, the receptor digest patterns were similar for estrogen receptors from the three different target cells . Our studies suggest considerable structural relatedness among these three estrogen receptors and also indicate that these two affinity labels bind to a similar, perhaps identical, region of the receptor molecule.

J Clin Pathol, 1988 Jun, 41(6), 671 - 5
Automated screening of blood cultures with the Malthus microbiological growth analyser; Brown DF et al.; A total of 3347 blood cultures from patients in all hospital wards were examined on a Malthus microbiological growth analyser and by a conventional system . There was no significant difference in the total numbers of positive cultures of clinical importance between the two systems (p greater than 0.05) . Staphylococcus aureus, however, was isolated more often by the conventional method (p less than 0.05) . Failure of the automatic detection routine limited the potential of the Malthus system for earlier detection of positive cultures . Daily visual examination of Malthus curves and subculture of bottles not promptly attached to the apparatus were necessary to avoid missing some positive cultures . False positive rates were 13% for the Malthus system and 2% for the conventional system . The contamination rate was considerably lower in the Malthus system (p less than 0.001) . Further development would be necessary for the apparatus to be acceptable for routine screening of blood cultures.

Proc Natl Acad Sci U S A, 1988 Jun, 85(11), 3975 - 9
Role for intracellular proteases in the processing and transport of class II HLA antigens; Blum JS et al.; Human B-lymphoblastoid cell lines (B-LCL) incubated with the protease inhibitor leupeptin accumulate complexes of class II HLA antigens with a series of Mr 21,000-23,000 basic proteins termed leupeptin-induced proteins (LIP) . The appearance of class II antigen-associated LIP coincides with the disappearance of class II antigen-associated invariant (I) chain . Glycopeptides generated by in vitro proteolysis of LIP and I chain using Staphylococcus aureus V8 protease are identical as determined by electrophoresis in sodium dodecyl sulfate . These results suggest that LIP is a proteolytic product derived from the I chain and are consistent with the view that further in vivo proteolysis of LIP by a leupeptin-sensitive enzyme normally facilitates its release from class II antigens . Incubation of B-LCL with monensin, which traps class II antigens and associated I chain in the Golgi apparatus, or chloroquine, which neutralizes intracellular acidic compartments and inhibits I-chain dissociation, blocks the leupeptin-induced appearance of LIP . Treatment of LIP with endoglycosidases F and H shows that both of its N-linked oligosaccharides are in the complex form, indicating that proteolysis of class II antigen-associated I chain to generate LIP occurs in a late-Golgi or post-Golgi compartment . The compartment in which these proteolytic events occur may be identical to the site in macrophages and B lymphocytes where foreign antigens are processed and interact with class II HLA molecules.

Pediatr Clin North Am, 1988 Jun, 35(3), 613 - 24
Methicillin-resistant Staphylococcus aureus: pediatric perspective; Kline MW et al.; Methicillin-resistant S . aureus has emerged as a nosocomial pathogen of major importance in pediatric patients . Infection occurs most often in hospitalized individuals with underlying predisposing medical conditions . Any body site may be involved, and bacteremia frequently occurs concomitantly . Vancomycin is the antibiotic of choice for serious MRSA infections; PRPs and cephalosporins generally are not effective . The likelihood of an adverse outcome of infection increases with the severity of an underlying condition and delay in institution of appropriate therapy . Infection control measures have met with only limited success in eradicating MRSA from the hospital environment . Methicillin-resistant S . aureus is likely to remain of considerable clinical significance to physicians caring for seriously ill children.

J Korean Med Sci, 1988 Jun, 3(2), 45 - 50
In vitro activities of eight antibiotics against methicillin-resistant S . aureus and S . epidermidis strains isolated in Korea; Chang WH et al.; Staphylococcus aureus and Staphylococcus epidermidis strains isolated at eight large medical centers in Korea were examined for methicillin resistance and resistance to eight other antibiotics; cefazolin, cefamandole, cefuroxime, cefoxitin, cefotaxime, moxalactam, penicillin G and vancomycin . Methicillin resistance was found in 296 of 1225 strains (24.2%) of S . aureus and 126 of 348 strains (36.2%) of S . epidermidis . Methicillinresistant strains were isolated from all sources with the frequency of isolation ranging from 11% to 60% . From pleural effusion, throat swab and blood, methicillin-resistant strains of S . aureus were more frequently isolated with statistical significance (Chi-squared test, 95% confidence) . Almost all of Methicillin-resistant S . aureus (MRSA) and S . epidermidis (MRSE) strains were multiply resistant to one or more tested eight antibiotics . However only 7(2.4%) of 296 MRSA strains and 2(1.6%) of 126 MRSE strains were resistant to vancomycin . Vancomycin was the most effective antibiotic against staphylococcal isolates as well as MRSA and MRSE.

Appl Environ Microbiol, 1988 Jun, 54(6), 1541 - 9
Plasmid profiles as indicators of the source of contamination of Staphylococcus aureus endemic within poultry processing plants; Dodd CE et al.; A total of 530 strains of Staphylococcus aureus were isolated from the defeathering machinery of a chicken processing plant and from neck skin samples of carcasses at different stages of processing in two visits 4 weeks apart . Eleven different plasmid profiles were detected in the isolates, eight being common to both visits . The plasmid profiles of the strains forming the majority of the population on the freshly slaughtered birds were rarely present in the strains isolated from the pluckers (except at the entry to the first plucker) and were present in only a small proportion of the strains isolated from carcasses after plucking . However, the profiles from the strains isolated from the pluckers on both visits were different from those forming the majority of the population on the incoming birds but formed the major part of the carcass flora after plucking, suggesting that such strains were endemic . These strains were found as a small proportion of the isolates made from the incoming birds, suggesting that this was the route by which the endemic strains were introduced into the plant . Such endemic strains exhibited a clumping growth, even in liquid shake culture, which may have made it easier for them to become established on the pluckers and to resist cleaning and disinfection . This clumping phenotype was correlated with the presence of a 7.5-megadalton plasmid.

Infect Immun, 1988 Jun, 56(6), 1470 - 4
Acidic fibroblast growth factor modulates Staphylococcus aureus adherence to human endothelial cells; Blumberg EA et al.; Alteration of human endothelial cells may increase their susceptibility to staphylococcal invasion and thus may contribute to the development of intravascular staphylococcal disease . Acidic fibroblast growth factor, a potent regulator of endothelial cell function, had a significant effect on Staphylococcus aureus infection of cultured human endothelial cells . Three of four S . aureus strains had diminished adherence to endothelial cells when the latter were grown in the presence of acidic fibroblast growth factor (P less than 0.05) . The diminished adherence was time dependent, maximal at 72 h, and independent of the initial bacterial inoculum . A twofold enhancement of S . aureus adherence was observed when endothelial cells were pretreated with heparitinase . Adherence was unaffected by endothelial cell activation by interleukin-1 or endotoxin . Thus, acidic fibroblast growth factor exerted a protective effect, deterring S . aureus adherence to cultured endothelial cells . Endothelial cell heparan sulfate was also directly involved in the adherence process . Subtle modulations of endothelial cells can significantly affect the ability of S . aureus to adhere to and then infect these cells . Similar alterations may contribute to the ability of S . aureus to infect endovascular tissue in vivo.

Southeast Asian J Trop Med Public Health, 1988 Jun, 19(2), 309 - 16
Childhood melioidosis in northeastern Thailand; Pongrithsukda V et al.; Eighteen cases of childhood melioidosis in Northeastern Thailand were reviewed . The mean age was 6.8 years with a range from eight months to 15 years . Twelve cases (66.7%) had localized melioidosis, six of which had pneumonia . Three patients were diagnosed as pharyngocervical melioidosis, the newly recognized syndrome . Nine cases (50.0%) had associated diseases including dengue hemorrhagic fever (DHF) in five cases . In all five cases, melioidosis was diagnosed during the convalescent stage as a cause of pyrexia with or without pneumonia . Pseudomonas pseudomallei strains isolated from 12 patients were all sensitive to chloramphenicol, cotrimoxazole and kanamycin . Ceftazidime, cefotaxime and ceftriaxone were also active against all six isolates tested . Three cases died, all were diagnosed as disseminated septicaemic melioidosis at postmortem . The overall mortality rate was 16.7% . The septicaemic form of melioidosis can resemble many diseases such as septicaemia due to Staphylococcus aureus or gram-negative organisms other than P . pseudomallei while the localized from may mimic pulmonary tuberculosis . A high index of clinical suspicion is required in making a diagnosis of melioidosis, particularly in areas where the disease is endemic.

Southeast Asian J Trop Med Public Health, 1988 Jun, 19(2), 175 - 85
Simple serological tests for detecting classical heat labile enterotoxin (LT-I) of Escherichia coli; Chaicumpa W et al.; Diarrhoea caused by enterotoxigenic Escherichia coli (ETEC) remains a problem in Southeast Asia . At present, no routine laboratories as yet are available for ETEC detection . In this study, attempts were made to produce reagents for use in simple serological tests for detecting LT . The serological methods were the Biken, the staphylococcal coagglutination and the reverse passive hemagglutination tests . For the Biken test, medium was prepared locally by mixing constituents as described previously by Honda et al., (1981) . Anti-CT-B subunit was prepared by immunizing a rabbit with commercial CT-B subunits (Sigma) . Other chemical reagents e.g . colistin, lincomycin etc . were obtained from the local supplies . Using the locally made reagents to detect LT from 100 WHO reference strains of E . coli by the Biken test, it was found that the test had 100%, 92%, 96%, 100% and 92.5% of specificity, sensitivity, accuracy, positive predictive value and negative predictive value, respectively . Protein A rich Staphylococcus aureus from the stock culture of the Department of Microbiology and Immunology, Faculty of Tropical Medicine, Mahidol University were grown in suitable medium i.e . blood agar containing lincomycin (BA-Lin) . Suitable amount of the rabbit anti CT-B subunit (0.1 ml) was used to sensitize each ml of the formalinized, heat-fixed bacteria . The sensitized bacteria were used for detecting LT in the lysates of the 100 E . coli reference strains . The lysates were prepared by growing the E . coli strains on BA-Lin medium for 8 hours, then a loopful of each strain was inoculated into colistin solution (20,000 unit/ml).(ABSTRACT TRUNCATED AT 250 WORDS)

J Gen Microbiol, 1988 Jun, 134 ( Pt 6), 1465 - 9
The expression in Staphylococcus aureus of cloned DNA encoding methicillin resistance; Inglis B et al.; A 4 kb fragment of chromosomal DNA was cloned from a clinical strain of methicillin-resistant Staphylococcus aureus . It comprises part of a section of the chromosome that was lost when the strain was cured of resistance to methicillin and to other antimicrobial agents . The fragment mediates an increased level of methicillin resistance when inserted into a shuttle vector and transformed back into the sensitive strain generated when the original DNA was deleted.

J Gen Microbiol, 1988 Jun, 134 ( Pt 6), 1455 - 64
Amplification of a section of chromosomal DNA in methicillin-resistant Staphylococcus aureus following growth in high concentrations of methicillin; Matthews PR et al.; Growth of two independently isolated strains of methicillin-resistant Staphylococcus aureus (MRSA) in increasing concentrations of methicillin (step-selection) resulted in increased resistance in these strains . When chromosomal DNA from the step-selected variants was probed using DNA sequences previously demonstrated to be associated with methicillin resistance in MRSA strains, amplification of the homologous chromosomal sequence was identified . Growth of these step-selected strains in the absence of methicillin resulted in loss of the amplified sequence, while the original sequence remained . There are differences between the two strains in the stability of maintenance of amplified sections . Prolonged storage of the variants on a high concentration of methicillin resulted in loss of amplified sections without concomitant loss of methicillin resistance . Thus amplification may be only one of at least two molecular mechanisms available to S . aureus to increase methicillin resistance in response to step-selection . Probing of cells of the highly resistant sub-population of a heterogeneously resistant MRSA strain showed that duplication of this mec-associated DNA is not involved in the mechanism of heteroresistance.

Microb Pathog, 1988 Jun, 4(6), 443 - 53
Cytotoxic effects of ingested Staphylococcus aureus on bovine endothelial cells: role of S . aureus alpha-hemolysin; Vann JM et al.; Previously we observed that Staphylococcus aureus phagocytized by cultured bovine endothelial cells do not proliferate intracellularly, but are cytotoxic to bovine endothelial cells . To investigate S . aureus virulence factors which may be produced intracellularly and cause lysis of endothelial cells, we tested S . aureus mutants defective in production of one or more potential virulence factors and corresponding parent strains for cytotoxicity to endothelial cell monolayers subsequent to being ingested . Following incubation of endothelial cell monolayers with S . aureus for 3.5 h, cultures were supplemented with lysostaphin to destroy extracellular but not intracellular S . aureus . At subsequent times, viability of endothelial cells was assayed by retention of 3H-adenine and the number of intracellular S . aureus was measured . The cytotoxic activity of S . aureus culture supernatants was also characterized . The results indicate that S . aureus alpha-hemolysin is cytotoxic to bovine endothelial cells and plays an important role in the damage suffered by bovine endothelial cell monolayers following ingestion of S . aureus . Ingestion of alpha-hemolysin-producing S . aureus by endothelial cells in vivo might be expected to result in destruction of endothelium followed by development of platelet-fibrin vegetations . This possible sequence of events is compatible with the frequently fulminant course of S . aureus endocarditis.

Antimicrob Agents Chemother, 1988 Jun, 32(6), 919 - 21
Efficacy of pefloxacin-fosfomycin in experimental endocarditis caused by methicillin-resistant Staphylococcus aureus; Thauvin C et al.; The efficacies of pefloxacin, fosfomycin, and both of these agents in combination against methicillin-resistant Staphylococcus aureus were assessed in a rat endocarditis model . The combination prevented emergence of the fosfomycin and pefloxacin resistance seen in 36 and 4%, respectively, of animals receiving either agent alone and was more effective than either agent in sterilizing cardiac vegetations.

Biochimie, 1988 Jun, 70(6), 783 - 9
Desmin heterogeneity in the main electric organ of Electrophorus electricus; Costa ML et al.; Desmin, the muscle-specific intermediate filament protein was purified from the main electric organ of Electrophorus electricus . It is shown that pure desmin can be separated into 5 isoforms presenting different isoelectric points . These isoforms have similar molecular weight, react with an antibody directed against desmin and generate identical peptides after digestion with protease V8 from Staphylococcus aureus.

Eur J Clin Microbiol Infect Dis, 1988 Jun, 7(3), 400 - 3
Occurrence of methicillin-resistant Staphylococcus aureus in a department of orthopedic surgery 1970 to 1986; Ipsen T et al.; The incidence of methicillin-resistant (and multiply-resistant) strains of Staphylococcus aureus in the Department of Orthopedic Surgery at Odense University Hospital fell from 15% of all strains in 1970 to less than 0.5% in 1986 . This decrease occurred more slowly in the orthopedic department than in the rest of the hospital . A retrospective survey of 30 episodes of infection due to methicillin-resistant Staphylococcus aureus from 1979 to 1986 indicates that this slower decrease may be due to the relatively frequent occurrence of chronic infections in orthopedic surgery.

Infect Control Hosp Epidemiol, 1988 Jun, 9(6), 255 - 60
Cost-effective eradication of an outbreak of methicillin-resistant Staphylococcus aureus in a community teaching hospital; Rao N et al.; During an eight-month period, 25 hospitalized patients became infected or colonized with methicillin-resistant Staphylococcus aureus (MRSA) in a 464-bed acute care, medical-surgical teaching hospital . There were only five cases during the eight months prior to the outbreak period (P less than 0.0001) . Initial measures, including category-specific isolation and education, did not limit the spread of the outbreak of a strain of MRSA . This prompted institution of additional measures including (1) strict isolation of all infected and colonized cases; (2) prospective microbiological surveillance to detect additional cases; (3) multiple site cultures of identified cases to determine the extent of colonization; (4) employee and environment surveillance; (5) antibiotic decolonization of patients and employees; and (6) educational efforts . The highest number of personnel carriers were noted in one of the critical care units where most of the cases occurred . The decolonization protocol was 100% effective for personnel carriers . The incidence of nosocomial cases of MRSA fell to zero in the five months following the implementation of the strategy . The cost of the entire eradication process was approximately half that of treating a single MRSA bacteremia.

J Biol Response Mod, 1988 Jun, 7(3), 283 - 95
Functional roles of gamma interferon in proliferation and differentiation of human B cell subpopulations; Xia X et al.; We have studied the function of interferon-gamma (IFN-gamma) on human B cell proliferation and differentiation . When B cell subpopulations were separated by Percoll gradient centrifugation and stimulated by Staphylococcus aureus Cowan I (SAC), these subpopulations responded differently to lymphokines . Small B cells (60/80% Percell) were stimulated to proliferate by IFN-gamma alone . Large B cells (50/60% Percoll) did not respond to IFN-gamma but proliferated in response to B cell growth factor (BCGF) free of interleukin-2 (IL-2) and IFN-gamma . Although IFN-gamma alone could not induce the differentiation of SAC-activated B cells and did not support the growth of large B cells, it enhanced the proliferation and differentiation of both subpopulations in the presence of BCGF and IL-2 . Also, IFN-gamma induced the expression of IL-2 receptor on B cells . Pretreatment of B cells with IFN-gamma for 48 h had a minor effect on the proliferation but significantly enhanced the differentiation in the presence of BCGF and IL-2; therefore, IFN-gamma may act as a differentiation factor . However, in a late stage of culture, IFN-gamma inhibited B cell differentiation . Our experimental data suggest that IFN-gamma is a growth factor for distinct subpopulation of SAC-activated human B cells and enhances the proliferation and differentiation and the expression of IL-2 receptor on B cells.

J Cell Biol, 1988 Jun, 106(6), 1927 - 36
Subcellular localization and quantitation of the major neutrophil pertussis toxin substrate, Gn; Bokoch GM et al.; The subcellular distribution of G protein subunits in the neutrophil was examined . Cells were nitrogen cavitated and subcellular organelles fractionated on discontinuous sucrose gradients . The presence of GTP-binding regulatory protein (G protein) alpha and beta/gamma subunits in each organelle was determined using three methods of analysis: specific binding of guanine nucleotide, ADP ribosylation by pertussis toxin, and immunoblot analysis with subunit-specific G protein antibodies . Both plasma membrane and cytosolic G protein components were detected . In contrast, neither the specific nor the azurophilic granules contained detectable G protein . Based on the ability of exogenous G protein beta/gamma subunits to increase the ADP ribosylation of the cytosolic form of G protein and upon the hydrodynamic behavior of the cytosolic protein, it is likely that this represents an uncomplexed G protein alpha subunit . Proteolytic mapping with Staphylococcus aureus V8 protease suggests the soluble alpha subunit is from Gn, the major pertussis toxin substrate of human neutrophils . Using quantitative analysis, the levels of the 40-kD G protein alpha subunit and of the 35/36-kD beta subunit in the neutrophil membrane were determined.

J Cell Sci, 1988 Jun, 90 ( Pt 2), 307 - 15
Spectrin-like proteins in the paraflagellar rod structure of Trypanosoma brucei; Schneider A et al.; A polyclonal, monospecific rabbit antibody to human erythrocyte spectrins cross-reacted with two sets of proteins (a doublet of 180/200K and a triplet of 67-66-65K; K = 10(3) Mr) in the parasitic protozoon Trypanosoma brucei brucei . Except for the 66K protein, the cross-reacting proteins are localized in the flagellum, on the basis of evidence from cell fractionation and immunofluorescence microscopy . Immunogold labelling and electron micrographs further revealed that the spectrin-like proteins are confined to the paraflagellar rod structure . The spectrin-like proteins with apparent molecular weights of 180 and 200 share homology with spectrin band 1, since V8-protease from Staphylococcus aureus generated similarly sized, antigenic peptides from these proteins . The results indicate homology between the cross-reacting proteins and human red cell spectrin.

Biokhimiia, 1988 Jun, 53(6), 918 - 24
{Determination of rate constants for the antigen-antibody reaction . Kinetic characteristics of interaction of soluble and corpuscular antigen with specific antibodies}; Bobrovnik SA et al.; Using the previously developed procedure, the rate and equilibrium constants for the interaction of antibodies with soluble or corpuscular antigens were determined . Cow milk casein was used as a soluble antigen, while heat-inactivated Staphylococcus aureus cells--as a corpuscular antigen . The values of kinetic constants for the above reactions are close to those for various antigen--antibody pairs obtained by other investigators.

Mol Biochem Parasitol, 1988 Jun, 29(2-3), 251 - 60
The 140/130/105 kilodalton protein complex in the rhoptries of Plasmodium falciparum consists of discrete polypeptides; Cooper JA et al.; Four monoclonal antibodies (MAbs) recognise an antigen localised in the rhoptries of Plasmodium falciparum merozoites using both indirect immunofluorescence assay and immunoelectron microscopy with immunogold labeling . All MAbs immunoprecipitated bands at 140, 130 and 105 kDa from {35S}methionine-labeled parasites; however, one MAb immunoblotted only the 130 kDa protein and another MAb immunoblotted the 105 kDa protein . The affinity purified antigen complex consisted of proteins of 140, 130, 105 and 98 kDa . The individual proteins were subjected to peptide mapping with Staphylococcus aureus V8 protease; the 98 kDa protein was a degradation product of the 105 kDa protein and the 140, 130, and 105 kDa proteins were found to be unrelated . The antigen complex was synthesised at the mid trophozoite stage and was considered to be soluble as judged by release from mature schizonts by freeze/thaw lysis . One of the MAbs inhibited parasite growth and/or merozoite invasion of erythrocytes, in vitro, to a small but significant extent.

Agents Actions, 1988 Jun, 24(1-2), 130 - 6
Enhanced antibacterial resistance in neutropenic mice treated with human recombinant interleukin-1 beta; Gladue R et al.; Human Recombinant IL-1 was investigated for its ability to increase non-specific resistance to Staphylococcus aureus in neutropenic mice . Mice, rendered neutropenic with cyclophosphamide and then administered IL-1 intraperitoneally, demonstrated enhanced resistance to subsequent challenge with S . aureus as measured by increased survival and bacterial clearance . No protective effects were observed with heat inactivated IL-1 . Efficacy was observed only when both IL-1 and the subsequent bacterial challenge were administered into the same site . Despite the observed protective effects, animals treated with IL-1 did not have increased numbers of blood leukocytes or peritoneal phagocytes prior to infection or at the times coincident with bacterial clearance . Based upon these observations, enhanced activity of resident macrophages may be responsible for the protective effects observed in IL-1 treated mice.

J Immunol, 1988 Jun 1, 140(11), 3736 - 44
T cell-dependent activation of B cell proliferation and differentiation by immobilized monoclonal antibodies to CD3; Hirohata S et al.; The capacity of mAb directed at the CD3 molecular complex (64.1) to induce T cell-dependent B cell proliferation and differentiation was examined . Coculture of B cells with mitomycin C-treated T4 cells (T4 mito) stimulated by immobilized 64.1 resulted in marked B cell proliferation and Ig-secreting cells (ISC) generation in the absence of any additional stimulation . The magnitude of the B cell responses induced by immobilized 64.1-stimulated T4 mito was far greater than that induced by other stimuli, such as Staphylococcus aureus plus factors produced by mitogen-activated T cells, PWM, or soluble 64.1 . The induction of maximal B cell responsiveness required direct contact between activated T cells and responding B cells . Of note, immobilized 64.1 also induced B cell proliferation and ISC generation in the presence of mitomycin C-treated T8 cells . By contrast, immobilized 64.1 stimulated T4 or T8 cells that had not been treated with mitomycin C induced very modest ISC generation and suppressed B cell responses supported by T4 mito even in the presence of exogenous IL-2 or factors produced by mitogen-activated T cells . The interactions between T and B cells in these cultures not only induced B cell responses, but also enhanced the production of IL-2 by activated T cells . Increased IL-2 production was facilitated when culture conditions afforded the opportunity for contact between B cells and activated T cells . These results indicate that the establishment of interactions between B cells and anti-CD3-stimulated T4 or T8 cells provides all of the signals necessary for proliferation and differentiation of B cells without other stimuli and also augments the production of lymphokines by the activated T cells . The data emphasize the role of Ag-nonspecific interactions between B cells and T cells in promoting polyclonal responses of both cell types.

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