Staphylococcus aureus

J Immunol, 1989 Mar 1, 142(5), 1569 - 75
Immunomodulatory role of IL-4 on the secretion of Ig by human B cells; Splawski JB et al.; The effect of IL-4 on the production of Ig by human B cells was examined . Highly purified B cells were stimulated with Staphylococcus aureus (SA) and IL-4 alone or in combination with various other cytokines and the supernatants assayed for Ig by isotype-specific ELISA . IL-4 (10 to 100 U/ml) did not support Ig secretion by SA-stimulated blood, spleen, or lymph node B cells, whereas IL-2 supported the production of all isotypes including IgE . Moreover, IL-4 suppressed the production of all isotypes of Ig by B cells stimulated with SA and IL-2 including IgG1, IgG2, and IgE . IL-4-mediated suppression was partially reversed by IFN-gamma or -alpha and low m.w . B cell growth factor . TNF-alpha and IL-6 did not reverse the IL-4-induced suppression of Ig production . The inhibitory action of IL-4 on Ig production appeared to depend on the polyclonal activator used to stimulate the B cells . Thus, Ig secretion by B cells activated by LPS and supported by IL-2 was not inhibited by IL-4 . Whereas IL-4 alone supported minimal Ig production by LPS-activated B cells, it augmented production of all Ig isotypes in cultures stimulated with LPS and supported by IL-2 . IFN-gamma further enhanced production of Ig in these cultures . When the effect of IL-4 on the responsiveness of B cells preactivated with SA and IL-2 was examined, it was found not to inhibit but rather to promote Ig production modestly . A direct effect of IL-4 on the terminal differentiation of B cells was demonstrated using B lymphoblastoid cell lines . IL-4 was able to enhance the Ig secreted by an IgA-secreting hybridoma, 219 and by SKW6-CL-4, an IL-6-responsive IgM-secreting EBV transformed B cell line . These results indicate that IL-4 exerts a number of immunoregulatory actions on human B cell differentiation . It interferes with the activation of B cells by SA and IL-2, but promotes the differentiation of preactivated B cells, B cell lines, and B cells activated by LPS without apparent isotype specificity.

J Gen Virol, 1989 Mar, 70 ( Pt 3), 647 - 58
Mapping of neutralizing epitopes to fragments of the bovine coronavirus E2 protein by proteolysis of antigen-antibody complexes; Deregt D et al.; Neutralizing antigenic domains on bovine coronavirus gp100/E2 were mapped to fragments of this protein by proteolytic cleavage and fragment analysis . The procedure involved analysis of fragments generated after incubation of E2-monoclonal antibody complexes with various proteases . The smallest antibody-bound fragments obtained were a 50K fragment following Staphylococcus aureus V8 protease and submaxillary protease digestion, and a 37K fragment following trypsin digestion . Trypsin also produced a transient antibody-bound 50K fragment . A 40K fragment which was not bound by antibody was observed following digestions with all three proteases . The 50K fragments generated by V8, submaxillary protease and trypsin comigrated on gels and displayed the same altered mobility under non-reducing conditions, suggesting identity of these fragments and indicating the presence of disulphide linkages in these fragments . The 40K fragments generated by these three enzymes also comigrated and displayed the same altered mobility under non-reducing conditions . The 37K trypsin fragment contained both neutralizing domains, A and B.

Laryngorhinootologie, 1989 Mar, 68(3), 169 - 80
{Antigen-specific antibody formation in determining the immune reaction in patients with head and neck cancers}; Wustrow TP; Head and neck cancer patients are considered to be immunodeficient . The immunological reaction, however, has been analysed only in unspecific systems or by cell surface markers . For the determination of the functional immunoreactivity of donors and patients with head and neck cancer the antigen-specific antibody production in vitro has been studied . Mononuclear cells from peripheral blood were stimulated with sheep red blood cells and after 5-7 days the antigen-specific immune response was measured in the presence of complement by red blood cell lysis . For maximal antibody production in vitro, the human lymphocytes were activated by Staphylococcus aureus and Interleukin-1 (IL-1) . The IL-1 concentration was measured by the proliferation of murine (C3H/HeJ) thymocytes . Without IL-1 and macrophages which had been separated by plastic adherence no antigen-specific antibody formation was observed . As the cultures were saturated with IL-1 and as the antigen-specific antibody production reached the same level even in the absence of macrophages, the antibody formation is not influenced by an altered macrophage function . Patients with head and neck cancer showed in vitro a significant reduction of the antigen-specific antibody production to no longer detectable values . In addition age-matched, healthy donors with high alcohol and cigarette abuse had a significant decrease in their antigen-specific antibody production compared to controls without high alcohol and cigarette consumption.

J Biol Chem, 1989 Feb 25, 264(6), 3629 - 35
Mechanism of the transition from plant ferritin to phytosiderin; Laulhere JP et al.; Soluble and insoluble forms of ferritins have been purified from dry pea seeds by gel filtration . The insoluble form is called phytosiderin by analogy with animal hemosiderin . Native gel electrophoresis of these two forms have shown that the soluble one (ferritin) is homogenous in size and more compact than the insoluble one (phytosiderin) which is heterogenous in size . However, when iron is removed from these two classes of molecules (apoferritin), they have the same mobility in isopycnic centrifugations . Polyacrylamide-sodium dodecyl sulfate gel electrophoresis revealed a difference in their subunit composition: ferritin molecules are built up from a 28-kDa subunit and phytosiderin from a 26.5-kDa subunit . Partial proteolysis using a Staphylococcus aureus protease indicates a strong relationship between these two polypeptides . Intermediates between these two forms have also been characterized and are composed of both subunits in various amounts . Ferritin and phytosiderin are both able to incorporate iron in vitro into their mineral core . It is also shown that in vitro iron exchange induces ferritin degradation . This degradation is prevented by inhibitors of the Fenton cycle (iron chelates like o-phenanthroline and desferrioxamine B) and reduced by Tris, a radical scavenger . Under in vitro conditions of controlled radical damage the 28-kDa subunit is converted into the 26.5-kDa subunit . Purification of the 28-kDa subunit has allowed us to determine the NH2-terminal sequence . The NH2 extremity of the 26.5-kDa subunit is heterogenous, but the sequence of its main component is identical to the sequence of the 28-kDa subunit downstream residue Leu-21 . These data indicate that the 26.5-kDa subunit is produced by radical mediated damage leading to a series of cleavages in the NH2 terminal part of the 28-kDa subunit.

Biochemistry, 1989 Feb 21, 28(4), 1574 - 81
Kinetics and specificity of reductive acylation of lipoyl domains from 2-oxo acid dehydrogenase multienzyme complexes; Graham LD et al.; Lipoamide and a peptide, Thr-Val-Glu-Gly-Asp-Lys-Ala-Ser-Met-Glu lipoylated on the N6-amino group of the lysine residue, were tested as substrates for reductive acetylation by the pyruvate decarboxylase (E1p) component of the pyruvate dehydrogenase multienzyme complex of Escherichia coli . The peptide has the same amino acid sequence as that surrounding the three lipoyllysine residues in the lipoate acetyltransferase (E2p) component of the native enzyme complex . Lipoamide was shown to be a very poor substrate, with a Km much higher than 4 mM and a value of kcat/Km of 1.5 M-1.s-1 . Under similar conditions, the three E2p lipoyl domains, excised from the pyruvate dehydrogenase complex by treatment with Staphylococcus aureus V8 proteinase, could be reductively acetylated by E1p much more readily, with a typical Km of approximately 26 microM and a typical kcat of approximately 0.8 s-1 . The value of kcat/Km for the lipoyl domains, approximately 3.0 x 10(4) M-1.s-1, is about 20,000 times higher than that for lipoamide as a substrate . This indicates the great improvement in the effectiveness of lipoic acid as a substrate for E1p that accompanies the attachment of the lipoyl group to a protein domain . The free E2o lipoyl domain was similarly found to be capable of being reductively succinylated by the 2-oxoglutarate decarboxylase (E1o) component of the 2-oxoglutarate dehydrogenase complex of E . coli . The 2-oxo acid dehydrogenase complexes are specific for their particular 2-oxo acid substrates . The specificity of the E1 components was found to extend also to the lipoyl domains.(ABSTRACT TRUNCATED AT 250 WORDS)

J Biol Chem, 1989 Feb 15, 264(5), 2560 - 7
The complete amino acid sequence of the low molecular weight cytosolic acid phosphatase; Camici G et al.; This paper presents the complete amino acid sequence of the low molecular weight acid phosphatase from bovine liver . This isoenzyme of the acid phosphatase family is located in the cytosol, is not inhibited by L-(+)-tartrate and fluoride ions, but is inhibited by sulfhydryl reagents . The enzyme consists of 157 amino acid residues, has an acetylated NH2 terminus, and has arginine as the COOH-terminal residue . All 8 half-cystine residues are in the free thiol form . The molecular weight calculated from the sequence is 17,953 . The sequence was determined by characterizing the peptides purified by reverse-phase high performance liquid chromatography from tryptic, thermolytic, peptic, Staphylococcus aureus protease, and chymotryptic digests of the carboxymethylated protein . No sequence homologies were found with the two known acylphosphatase isoenzymes or the metalloproteins porcine uteroferrin and purple acid phosphatase from bovine spleen (both of which have acid phosphatase activity) . Two half-cystines at or near the active site were identified through the reaction of the enzyme with {14C} iodoacetate in the presence or in the absence of a competitive inhibitor (i.e . inorganic phosphate) . Ac-A E Q V T K S V L F V C L G N I C R S P I A E A V F R K L V T D Q N I S D N W V I D S G A V S D W N V G R S P N P R A V S C L R N H G I N T A H K A R Q V T K E D F V T F D Y I L C M D E S N L R D L N R K S N Q V K N C R A K I E L L G S Y D P Q K Q L I I E D P Y Y G N D A D F E T V Y Q Q C V R C C R A F L E K V R-OH.

Biochem J, 1989 Feb 15, 258(1), 205 - 9
Clavulanate inactivation of Staphylococcus aureus beta-lactamase; Rizwi I et al.; The interaction of clavulanic acid with beta-lactamase from Staphylococcus aureus was investigated, particularly with a view to determining whether conformational effects are involved . The inactivation at neutral pH is essentially stoichiometric, leading to an inactive species with an enamine chromophore . Two forms of the enamine were observed, the first-formed having a positive ellipticity with a maximum near 290 nm . This species slowly converted into the stable form of the inactivated enzyme that had a negative ellipticity with a minimum at 275 nm . This change in sign of the ellipticity of the enamine is consistent with the previously proposed cis-trans isomerization of the enamine {Cartwright & Coulson (1979) Nature (London) 278, 360-361) . Both the far-u.v.c.d . and the intrinsic viscosity of the inactivated enzyme indicated that negligible change in conformation of the enzyme accompanied inactivation . The rates of inactivation and enamine formation were compared at low temperatures, where the initial rates were slow enough to be monitored . The rate of loss of 95% of the catalytic activity was almost 100-fold faster than the rate of formation of the first-formed enamine species . The remaining 5% activity was lost with a rate comparable with that for formation of the initial enamine . The simplest explanation of these results is that a relatively stable acyl-enzyme intermediate builds up initially and more slowly partitions between turnover (hydrolysis) and enamine formation . The initially formed enamine is in the cis conformation but slowly isomerizes to the more stable trans form.

J Immunol Methods, 1989 Feb 8, 117(1), 53 - 8
Simultaneous measurement by flow cytometry of phagocytosis and hydrogen peroxide production of neutrophils in whole blood; Hasui M et al.; Bacterial ingestion and hydrogen peroxide production by polymorphonuclear leukocytes (PMN) were simultaneously measured using flow cytometry and 2',7'-dichlorofluorescein diacetate and fluorescent Staphylococcus aureus . Only 100 microliters of a whole blood specimen were required for these determinations, and the results were found to be independent of the absolute numbers of PMN, making the purification and adjustment of PMN numbers unnecessary . A positive correlation between phagocytosis and hydrogen peroxide production by individual PMN was demonstrated in healthy adults.

Biochemistry, 1989 Feb 7, 28(3), 964 - 8
Structure of polyubiquitinated histone H2A; Nickel BE et al.; We have recently demonstrated that trout liver histones H2A, H2B, and H2A.Z can be polyubiquitinated {Davie, J.R., Delcuve, G.P., Nickel, B.E., Moyer, R., & Bailey, G . (1987) Cancer Res . 47, 5407-5410} . In the present study we determined the arrangement of the ubiquitin molecules in polyubiquitinated histone H2A . Trout liver chromatin fragments . which had histone H1 removed, were digested with Staphylococcus aureus (V8 strain) protease which cleaves specifically on the carboxyl side of glutamic acid residues under the conditions used . The V8 protease readily degraded histone H2A and ubiquitinated (u) H2A at equivalent rates . One site in H2A and uH2A, the peptide bond between Glu 121 and Lys 122, was cleaved, yielding protein species cH2A and cuH2A, respectively . None of the other nucleosomal histones (H2B, H2A.Z, H3, and H4) including uH2B and uH2A.Z were sensitive to digestion . Trout liver histones cleaved with either V8 protease, histone H2A specific protease, or cyanogen bromide were resolved by two-dimensional gel electrophoresis and ubiquitinated peptides detected with anti-ubiquitin IgG . The results suggest that the major arrangement of ubiquitin in polyubiquitinated H2A is a chain of ubiquitin molecules joined to each other by isopeptide bonds to a ubiquitin molecule that is attached to the epsilon-amino group of lysine 119 of histone H2A.

Acta Orthop Scand, 1989 Feb, 60(1), 113 - 5
Antibiotic treatment insufficient for established septic arthritis . Staphylococcus aureus experiments in rabbits; Riegels-Nielsen P et al.; We treated septic arthritis of the knee in 38 rabbits with cloxacillin i.m . once and twice daily combined with probenecid for 7 or 21 days, respectively, or with only cloxacillin i.m . thrice daily for 7 days . The animals were killed weekly in groups up to 5 weeks after inoculation . Aspirated cultures obtained after 4 days of treatment were always negative . Histologic specimens revealed progressive joint destruction, but at a slower rate after frequent treatment independent of the period . We concluded that antibiotic therapy alone could not prevent destruction of articular cartilage once bacterial arthritis was established.

Ther Umsch, 1989 Feb, 46(2), 111 - 7
{Atopic neurodermatitis}; Thurlimann W; Neurodermatitis atopica is a skin disease caused by various different factors and which is characterized by the simultaneous presence of different clinical features . The treatment is directed towards the various aetiopathological factors and oriented on the clinical picture . The main symptom is dry skin, which has to be rehydrated and provided with an artificial film of grease . Dermosteroids are the most efficient antiinflammatory agents, but they can only be used as one of various components of a complete neurodermatitis therapy . New knowledge gained over the past few years has shown the important role of Staphylococcus aureus in this condition, and as a result antibiotic therapy has increased in importance . The use of soap, which was previously not recommended, is now considered to be a useful and beneficial addition to the treatment, thanks to the availability of new alkali-free, disinfectant soaps . In certain cases there is a causal connection with exposure to allergens . Besides treatment of the itching, which is crucial for the patient, stabilization of the psychological factor and, if necessary, changes in the patient's environmental situation are extremely important . Substitution of essential fatty acids and UV therapy are discussed.

J Antibiot (Tokyo), 1989 Feb, 42(2), 198 - 205
Fosfonochlorin, a new antibiotic with spheroplast forming activity; Takeuchi M et al.; A new antibiotic, fosfonochlorin, was found in the culture filtrate of four strains of fungi freshly isolated from soil samples . These strains were identified as Fusarium avenaceum, Fusarium oxysporum, Fusarium tricinctum and Talaromyces flavus . Fosfonochlorin was a low molecular weight antibiotic (MW 158), soluble in water and methanol, but insoluble in acetone, ethyl acetate and chloroform . It was named after its possession of phosphorus and chlorine atoms, each one molar in its structure . The structure was determined as chloroacetylphosphonic acid mainly by the 1H NMR and mass spectrometric analyses . It was moderately active against some species of Gram-negative bacteria and its synergistic effect with glucose-6-phosphate was observed on Staphylococcus aureus and Escherichia coli . Spheroplast formation of the susceptible organisms with this antibiotic suggested that it might inhibit their cell wall synthesis.

APMIS, 1989 Feb, 97(2), 166 - 74
Generation and characterization of monoclonal antibodies against Staphylococcus aureus thermonuclease; Brakstad OG et al.; Monoclonal antibodies (MAbs) against Staphylococcus aureus thermonuclease (TN) were raised by immunizing BALB/c mice with a commercial TN preparation . Six monoclones were generated producing MAbs specific for S . aureus TN as tested in Western blots and ELISA . They all combined with a 17 kD and a 21 kD protein, respectively, both of which showed DNase activity . All MAbs were of IgG1 isotype with kappa light chain . Competition ELISA showed that five of the MAbs recognized a total of three different binding sites of TN, designated I, II and III, respectively . Only the anti-site II MAbs inhibited the DNase activity . A MAb-based sandwich ELISA showed a lower detection limit for TN of approximately 0.5 ng/ml protein . Only S . aureus strains (culture supernatants) showed positive ELISA (31 positive/31 tested), although other tested gram positive cocci produced thermostable nucleases . The MAbs have potentials as reagents for rapid and specific detection of S . aureus.

South Med J, 1989 Feb, 82(2), 165 - 8
Evaluation of antibacterial sensitivity testing methods for methicillin-resistant Staphylococcus aureus in a dermatology outpatient population; McBride ME et al.; Over a period of one year, 1986-1987, 116 strains of Staphylococcus aureus were isolated from patients attending two outpatient dermatology clinics in Houston, Texas . The purpose of this study was to evaluate the adequacy of routine antibiotic sensitivity testing methods for detecting methicillin-resistant Staphylococcus aureus (MRSA) . The Kirby-Bauer disk diffusion method was compared with a commercially available screening medium containing 6 micrograms/ml of oxacillin and 4% NaCl . The minimal inhibitory concentration (MIC) of methicillin, oxacillin, and oxacillin with 4% NaCl to S aureus using the agar dilution method was also determined . Approximately 90% of S aureus strains produced beta-lactamase and were resistant to penicillin and ampicillin . By disk diffusion, no strains were resistant to methicillin, though diameters of zones of inhibition were between 10 and 14 mm in seven strains . All strains proved to be sensitive to methicillin by MIC determinations and on the oxacillin-NaCl screening medium . The MIC of methicillin was 2.5 micrograms/ml for the majority of strains of S aureus, between 0.16 and 0.31 microgram/ml for oxacillin, and 0.08 to 0.16 microgram for oxacillin with 4% NaCl . We concluded that the incidence of MRSA in an outpatient dermatology population is low, and a combination of disk diffusion and oxacillin-NaCl screening is adequate for testing sensitivity.

Neurology, 1989 Feb, 39(2 Pt 1), 173 - 8
Neurologic complications of endocarditis: a 12-year experience; Salgado AV et al.; We reviewed the neurologic complications in 113 patients with native and 62 patients with prosthetic valve endocarditis . Neurologic complications occurred with the same frequency (35.3% vs 38.7%) and distribution among the two groups . Death occurred in 20.6% of patients with neurologic complications and in 13.6% of patients without neurologic complications (p = 0.23) . Staphylococcus aureus endocarditis correlated statistically with the development of neurologic complications (p less than 0.01) and death (p less than 0.01) . Among 50 patients discharged from the hospital after receiving only medical treatment for native valve endocarditis, and followed for a mean period of 48 months, there was one patient with mitral valve prolapse and stroke . We conclude that (1) neurologic complications occur with the same frequency in native and prosthetic valve endocarditis, (2) S aureus endocarditis increases the risk of neurologic complications and death, (3) mortality is not significantly increased in patients with neurologic complications, and (4) an episode of treated native valve endocarditis does not increase the natural history of stroke in valvular disease.

J Med Microbiol, 1989 Feb, 28(2), 129 - 36
Protection of mice by a pseudodiffuse strain of Staphylococcus aureus possessing polyvalent capsular type antigen; Chomarat M et al.; Staphylococcus aureus strain MC31 showed pseudodiffuse growth in serum-soft agar and reacted with immune rabbit sera to strains Smith diffuse (capsular type A), NS58D (type B) and NS41D (type C) but not with strain NS68D (type D) in serum-soft agar . Immunisation of mice with 300 micrograms of cell-surface polysaccharide extracted from strain MC31 protected against lethal infection by strain MC31 and the strains of capsular types A, B and C . Immune rabbit serum prepared against strain MC31 passively protected mice against challenge infection with the homologous strain, but approximately 30 times more anti-MC31 serum was required to protect against infection with the strains of capsular types A, B and C . Absorption of the passive protective activity of immune sera raised against the three capsular type strains required at least 10 times the quantity of MC31 cell-surface polysaccharide than the quantity of cell-surface polysaccharide from the homologous capsular strain . Electronmicrographs of strain MC31 treated with ferritin-labelled antisera to the three capsular strains showed only small amounts of ferritin granules around the cell wall.

Surg Gynecol Obstet, 1989 Feb, 168(2), 138 - 42
An experimental study of susceptibility to infection after hemorrhagic shock; Livingston DH et al.; Hemorrhagic shock has been associated with an increased risk of infection after injury . The immediate and long term effects of hemorrhagic shock without tissue injury on the susceptibility of an animal to infection and the efficacy of antibiotic prophylaxis to prevent infection in this setting were examined . Sprague-Dawley rats were subjected to hemorrhagic shock (LD15) by bleeding to a mean arterial pressure of 45 millimeters of mercury for 45 minutes and were resuscitated with shed blood and normal saline solution . In one experiment, dorsal wounds were inoculated one hour before or after shock with either 10(6), 10(8) or 10(10) Staphylococcus aureus . In a second experiment, rats were infected at one hour, or one, three or five days after shock with 10(6), 10(7) or 10(8) S . aureus . Equivalent numbers of rats received cefamandole nafate prior to bacterial challenge . Hemorrhagic shock increased the susceptibility to wound infection at all inocula . Infection increased whether rats were wounded before or after shock, and this effect was sustained for up to three days . Antibiotic prophylaxis was of limited value in reducing the incidence of wound infection after shock.

J Virol, 1989 Feb, 63(2), 647 - 58
Characterization of human immunodeficiency virus type 2 envelope glycoproteins: dimerization of the glycoprotein precursor during processing; Rey MA et al.; Four glycoproteins with apparent molecular weights of 300,000, 140,000, 125,000, and 36,000 (gp300, gp140, gp125, and gp36) were detectable in human immunodeficiency virus type 2 (HIV-2)-infected cells . gp125 and gp36 are the external and transmembrane components, respectively, of the envelope glycoproteins of HIV-2 mature virions . gp300 and gp140 are only detectable in virus-infected cells . They have identical isoelectric points, suggesting that gp300 might be a dimeric form of the immature precursor, gp140 . The purified gp300 can be dissociated in a slightly acidic buffer to give rise to monomers of 140,000 molecular weight . Such dissociated monomers and the purified gp140 showed identical patterns of polypeptides after partial proteolysis with Staphylococcus aureus V8 protease . Pulse-chase experiments indicated that gp300 is formed after synthesis of gp140 and before the detection of the mature external envelope glycoprotein, gp125 . These results were confirmed by using various inhibitors of glycosylation and inhibitors of trimming enzymes . Dimer formation of the envelope glycoprotein precursor was also observed in cells infected with simian immunodeficiency virus (SIV), a virus closely related to HIV-2 . On the other hand, the envelope glycoprotein precursor of HIV-1 did not form a dimer during its processing . Therefore, dimer formation seems to be a specific property of HIV-2 and SIV envelope gene expression . Such transient dimerization of the glycoprotein precursor might be required for its efficient transport to the Golgi apparatus and for its processing.

Cent Afr J Med, 1989 Feb, 35(2), 327 - 9
Antibiotic resistance of Staphylococcus aureus isolates from community acquired soft tissue infections in Bulawayo; Oliver MJ et al.; Fifty cases of community acquired strains of Staphylococcus aureus causing superficial abscesses were tested for their sensitivity to commonly used antibiotics . Ninety-two percent of isolates were resistant to benzyl-penicillin and ampicillin . Possible causes for this very high level of resistance and the implications for antibiotic usage are discussed.

Eur J Cell Biol, 1989 Feb, 48(1), 19 - 26
A colloidal gold labeling technique for the direct determination of the surface area of eukaryotic cells; Kehle T et al.; We have developed a colloidal gold labeling technique for the direct quantitation of the cell surface area . The method is based on coating the cell surface with {195Au} colloidal gold-protein complexes followed by morphometric determination of the labeling density (gold particles/micron2 cell surface) and radiometric determination of the total number of gold particles bound per cell . The ratio of both values directly gives the cell surface area . The accuracy of the method was shown using Staphylococcus aureus cells as a model system, where the cell surface area determined with our assay (4.0 microns2) corresponded well to the value calculated from the radius of the cells (3.6 microns2) . In a more complex model system J-774 mouse macrophages were labeled with different amounts of {195Au} gold-protein complexes to show that the assay is independent of the degree of saturation of the cell surface binding sites . Both high (135 Au/microns2) and low (65 Au/microns2) labeling densities resulted in a surface area of about 1200 microns2 . The technique finally was applied to L-929 fibroblasts to determine the increase of the cell surface area when the cells change from a spherical to a flat monolayer state . We found that the cell surface area increased 3-fold during the spreading process . The results show that the colloidal gold labeling technique allows the direct determination of the surface area of complex eukaryotic cells . The technique is suitable for the quantitation of changes in the surface architecture known to occur in different functional states of eukaryotic cells.

Kyobu Geka, 1989 Feb, 42(2), 160 - 3
{A case report of tricuspid valve endocarditis due to Staphylococcus aureus with successful surgical treatment}; Takayama T et al.; A 14-year-old boy had previously received right temporal lobectomy under the diagnosis of a malignant brain tumor . About one month after lobectomy, ventriculo-peritoneum shunt and ventriculo-atrium shunt were placed because hydrocephalus was progressed . The patient subsequently had a high fever probably due to wound infection of the shunt operation . Several blood cultures demonstrated Methicillin Resistant Staphylococcus Aureus (MRSA) . Several sensitive antibiotics were administered for about 30 days, however these drugs were not effective . According to echocardiography, moderate tricuspid regurgitation and a large vegetation at the tricuspid valve were detected and isolated tricuspid valve endocarditis was diagnosed . Surgical intervention was necessary because of recurrent pulmonary emboli . After tricuspid valve replacement with a Bjork-Shiley mechanical valve (31 mm), fever subsided and the patient was discharged on the 38th postoperative day . It is concluded that the surgical indications of the tricuspid valve endocarditis are as follows: 1 . recurrent pulmonary emboli, 2 . refractory right heart failure, 3 . resistance against antibiotics.

Electrophoresis, 1989 Feb, 10(2), 152 - 7
Development of a database of amino acid sequences for proteins identified and isolated on two-dimensional polyacrylamide gels; Sweatt JD et al.; As part of our continuing studies into the biochemical basis of long-term changes in neuronal function in Aplysia, we have developed a simple method for obtaining amino acid sequence information from proteins isolated on two-dimensional gels . Proteins isolated on preparative two-dimensional gels are digested in situ with Staphylococcus aureus V8 protease, and the resulting peptides electrophoresed, transferred to a polyvinylidene difluoride membrane, and sequenced in a gas-phase sequencer . The method is simple and should be applicable to a variety of other systems where the development of a two-dimensional gel database is underway.

Zh Mikrobiol Epidemiol Immunobiol, 1989 Feb, (2), 68 - 71
{Features of contamination of the environment by Staphylococcus aureus carriers}; Riabinin NV; The specific features of the spread of S . aureus from carriers indoors were studied . In this study S . aureus cells were most frequently isolated from the floor, less frequently from the walls and furniture and even less frequently from the air . S . aureus were detected on environmental objects as early as 30 minutes after the carrier stayed and breathed in the room . The contamination of environmental objects increased when the carrier spoke loudly and made such respiratory acts as coughing and sneezing . After the carrier left the room the contamination of the environmental objects persisted for at least 14 hours (the term of observation), gradually decreasing . The degree of the contamination of environmental objects was inversely related to the distance of these objects from the carrier . The contamination of the environment was shown to be directly related to the concentration of S . aureus in the nasal cavity of carriers.

Antimicrob Agents Chemother, 1989 Feb, 33(2), 181 - 4
Ciprofloxacin therapy for methicillin-resistant Staphylococcus aureus infections or colonizations; Smith SM et al.; Thirty patients were treated for colonization or for skin and soft tissue infections caused by methicillin-resistant Staphylococcus aureus . Three treatment regimens were evaluated, each progressively more aggressive . Our regimen was 750 mg of ciprofloxacin twice daily for 5 days, the second regimen was 750 mg of ciprofloxacin twice daily for 10 to 14 days, and the final regimen was 750 mg of ciprofloxacin twice daily plus 300 mg of rifampin twice daily for 21 days . It appears that ciprofloxacin alone produced an initial eradication rate in at least one site in 50% of the patients, regardless of whether the treatment was for 5 or up to 14 days . All of the patients with eradication became recolonized within 1 week posttherapy . When rifampin was combined with ciprofloxacin, the eradication rate was 100% when the isolates were susceptible to both agents, and these patients remained free of methicillin-resistant S . aureus at 1-week and 1-month follow-ups.

Harefuah, 1989 Feb 1, 116(3), 145 - 6
{Nontropical pyomyositis}; Klein L et al.; Pyomyositis of the pectoralis major was diagnosed in a 79-year-old man and Staphylococcus aureus was grown from pus drained from the infected muscle . Bacteremia and overwhelming sepsis accompanied the infection, and the patient died despite early diagnosis, combined chemotherapy and surgical drainage . The incidence of pyomyositis has been increasing lately in temperate climates . To our knowledge, this is the first report of nontropical pyomyositis in Israel.

J Appl Bacteriol, 1989 Feb, 66(2), 153 - 9
Differentiation of Staphylococcus aureus from freshly slaughtered poultry and strains 'endemic' to processing plants by biochemical and physiological tests; Mead GC et al.; A comparison was made of 27 'endemic' strains of Staphylococcus aureus and 35 strains from freshly slaughtered birds, isolated at five commercial slaughterhouses processing chickens or turkeys . Of 112 biochemical and physiological tests used, 74 gave results which differed among the strains . Cluster analysis revealed several distinct groupings which were influenced by strain type, processing plant and bird origin; these included a single group at the 72% level of similarity containing most of the 'endemic' strains . In comparison with strains from freshly slaughtered birds, a higher proportion of 'endemic' strains produced fibrinolysin, alpha-glucosidase and urease and were beta-haemolytic on sheep-blood agar . The 'endemic' type also showed a greater tendency to coagulate human but not bovine plasma, and to produce mucoid growth and clumping . The last two properties, relevant to colonization of processing equipment, were less evident in heart infusion broth than in richer media or process water collected during defeathering of the birds.

Gen Comp Endocrinol, 1989 Feb, 73(2), 252 - 9
The complete amino acid sequence of growth hormone of an elasmobranch, the blue shark (Prionace glauca); Yamaguchi K et al.; The complete amino acid sequence of growth hormone (GH) from a phylogenetically ancient fish, the blue shark (Prionace glauca), was determined . The shark GH isolated from pituitary glands by U . J . Lewis, R . N . P . Singh, B . K . Seavey, R . Lasker, and G . E . Pickford (1972, Fish . Bull . 70, 933-939) was purified by reversed-phase high-performance liquid chromatography . The hormone was reduced, carboxymethylated, and subsequently cleaved in turn with cyanogen bromide and Staphylococcus aureus protease . The intact protein was also cleaved with lysyl endopeptidase and o-iodosobenzoic acid . The resulting peptide fragments were separated by rpHPLC and submitted to sequence analysis by automated and manual Edman methods . The shark GH consists of 183 amino acid residues with a calculated molecular weight of 21,081 . Sequence comparisons revealed that the elasmobranch GH is considerably more similar to tetrapod GHs (e.g., 68% identity with sea turtle GH, 63% with chicken GH, and 58% with ovine GH) than teleostean GHs (e.g., 38% identities with salmon GH and 42% with bonito GH) except for eel GH (61% identity), and substantiates the earlier finding derived from the immunochemical and biological studies (Hayashida and Lewis, 1978) that the primitive fish are less diverged from the main line of vertebrate evolution leading to the tetrapod than are the modern bony fish.

Gen Comp Endocrinol, 1989 Feb, 73(2), 242 - 51
The complete amino acid sequence of growth hormone from the sea turtle (Chelonia mydas); Yasuda A et al.; The complete amino acid sequence of growth hormone (GH) from a reptilian species (the sea turtle, Chelonia mydas) has been determined for the first time . The hormone was reduced, carboxymethylated, and subsequently cleaved in turn with cyanogen bromide and Staphylococcus aureus protease . The intact protein was also cleaved with lysyl endopeptidase and o-iodosobenzoic acid . The resulting fragments were exclusively separated by reversed-phase high-performance liquid chromatography and subjected to sequence analysis by automated gas-phase sequencer employing the Edman method . The sea turtle GH consist of 190 amino acid residues with two disulfide linkages formed between residues 52-160 and 180-188, and possesses a microheterogeneity, indicated by the presence or absence of an additional alanine residue at the N-terminus . Sequence identities of sea turtle GH to other species of GH are 89% with chicken GH, 79% with rat GH, 68% with blue shark GH, 58% with eel GH, 59% with human GH, and 40% with a teleostean GH such as chum salmon . On the basis of amino acid sequence comparisons, a molecular phylogenetic tree is proposed.

J Dairy Sci, 1989 Feb, 72(2), 540 - 4
Effect of segregation on prevention of intramammary infections by Staphylococcus aureus; Fox LK et al.; Effectiveness of segregating cows with Staphylococcus aureus intramammary infections was studied over 1 yr . Nine herds were split into control (n = 5) or segregated (n = 4) groups . Cows with S . aureus intramammary infections were milked last in segregated herds . Monthly milk samples were collected aseptically for microbiologic analysis . Mean incidences of S . aureus intramammary infections were 3.7 and 4.3 cases/100 cow-mo in segregated and control herds . The mean prevalence of S . aureus intramammary infections decreased in both segregated and control herds during the study . Mean percentages of cows with S . aureus intramammary infections at the beginning and end of the study were 33.7 and 21.5 in segregated herds and 25.3 and 15.0 in control herds . Cows in all herds with S . aureus intramammary infections were preferentially culled . There were no significant differences in incidence and prevalence of S . aureus intramammary infections between groups, suggesting that S . aureus intramammary infections can be controlled without segregation.

Diabetes Care, 1989 Feb, 12(2), 153 - 5
Carriage of Staphylococcus aureus and inflamed infusion sites with insulin-pump therapy; van Faassen I et al.; The most common complication of continuous subcutaneous insulin infusion (CSII) is inflammation at the infusion site . To determine possible risk factors to these infections, we studied several factors in the management of CSII and compared the pyogenic skin inflammation rate, the carriage rate of Staphylococcus aureus, and the HbA1 level among 50 CSII-treated diabetic patients, 50 diabetic patients on insulin injections, 48 diabetic patients on oral medication, and 40 healthy volunteers . There was no increased carriage rate of S . aureus among CSII-treated patients (42%) as compared with the other groups . An unexpected inverse relationship existed between HbA1 level and carriage rate in the CSII-treated group (HbA1 5-8%, n = 16, 69%; HbA1 8-10% n = 15, 40%; HbA1 greater than 10, n = 19, 21% P = .02) . Pyogenic skin inflammations were reported by 24 (48%) CSII-treated patients, of which 18 had infected infusion sites, 6 (12%) insulin injecting patients, 2 (4%) patients on oral medication, and 3 (8%) healthy volunteers (P less than .01) . The occurrence of inflamed infusion sites was not associated with carriage of S . aureus, the indwelling time of the needle, or the insulin dosage per day . There was an association, however, with the type of insulin preparation classified according to the added preservative: m-cresol-containing insulin (n = 24, 54%); methyl p-hydroxybenzoate-containing insulin (n = 26, 19%, P = .02) . We concluded that the carriage of S . aureus is not increased among diabetic patients on CSII treatment and is not a risk factor in the occurrence of inflammation at the infusion site.(ABSTRACT TRUNCATED AT 250 WORDS)

Cutis, 1989 Feb, 43(2), 140 - 2
Botryomycosis caused by Moraxella nonliquefaciens; Feldman SR et al.; Botryomycosis is a bacterial infection characterized by the presence of grains of organisms in the tissue . Although Staphylococcus aureus is the most common causative agent, other bacteria have been reported to cause botryomycosis . Several factors have been hypothesized to be important in the pathogenesis of botryomycosis, including foreign bodies, quantity and virulence of organisms, and host immunity . The resulting infection tends to be resistant to antibiotic therapy . We report a case of botryomycosis of the hand caused by Moraxella nonliquefaciens, an organism of low virulence not previously associated with botryomycosis, and discuss the factors that may have led to this infection . This infection was successfully treated by excision and grafting.

Appl Environ Microbiol, 1989 Feb, 55(2), 523 - 5
Cell-free mercury volatilization activity from three marine caulobacter strains; Ji GY et al.; Three mercury-resistant marine Caulobacter strains showed an inducible mercury volatilization activity . Cell-free mercury volatilization (mercuric reductase) from these three marine Caulobacter strains was characterized and compared with enzyme activities determined by plasmids of Escherichia coli and Staphylococcus aureus . The temperature sensitivity of the Caulobacter mercuric reductase was greater than that of mercuric reductase from other gram-negative sources . Cell-free enzyme activity required NADH or NADPH, with NADPH functioning much better at lower concentrations than NADH . The Km for the Caulobacter enzyme was 4 microM Hg2+ . Ag+ was a competitive inhibitor of Caulobacter mercuric reductase (Ki = 0.2 microM Ag+), as with previously studied enzymes . Arsenite was a noncompetitive inhibitor of the Caulobacter enzyme with a Ki of 75 microM AsO2-.

Transplantation, 1989 Feb, 47(2), 300 - 4
The effect of exogenous lymphokines on immunoglobulin production and lymphocyte proliferation in renal transplant patients; Cardella CJ et al.; Peripheral blood mononuclear cells from stable long-term kidney transplant patients were activated in vitro by Staphylococcus Aureus Cowan I (SAC) (a B cell mitogen) . The effect of exogenous interleukin 2 and/or B cell growth factor (BCGF) on these cells was measured by 3H-thymidine incorporation and immunoglobulin production . Both proliferation and Ig production were lower in SAC-activated cells from transplanted patients compared to controls (P less than 0.01) . BCGF significantly enhanced blastogenesis (P less than 0.01) and Ig production (P less than 0.01) in SAC-treated cells from either patients or controls; however, the SAC- and BCGF-treated cells of transplant patients did not reach normal proliferation or immunoglobulin production values (P less than 0.001, P less than 0.01, respectively) . The addition of IL-2 to SAC-activated cells also increased proliferation and Ig production both in controls (P less than 0.05) and patients (P less than 0.005) . However, cells from transplant patients treated with SAC and IL-2 did not reach the normal levels of proliferation or immunoglobulin production (P less than 0.05 for both) . IL-2 did not enhance the increase of immunoglobulin production brought about by BCGF . SAC-activated B cells from transplant patients do not proliferate normally and do not produce normal amounts of Ig . The addition of IL-2 and BCGF results in a partial but subnormal improvement in both proliferation and Ig production . We conclude that the B cell abnormality in transplant patients may be due to lack of T cell lymphokines and an intrinsic B cell defect . These results suggest that the administration of exogenous lymphokines to transplant patients with B cell dysfunction may be clinically useful.

J Bacteriol, 1989 Feb, 171(2), 684 - 91
Identification and cloning of the conjugative transfer region of Staphylococcus aureus plasmid pGO1; Thomas WD Jr et al.; The conjugative transfer (tra) genes of a 52-kilobase (kb) staphylococcal plasmid, pGO1, were localized by deletion analysis and transposon insertional inactivation . All transfer-defective (Tra-) deletions and Tn551 or Tn917 transposon insertions occurred within a 14.5-kb BglII fragment . Deletions and insertions outside this fragment all left the plasmid transfer proficient (Tra+) . The tra region was found to be flanked by directly repeated DNA sequences, approximately 900 base pairs in length, at either end . Clones containing the 14.5-kb BglII fragment (pGO200) and subclones from this fragment were constructed in Escherichia coli on shuttle plasmids and introduced into Staphylococcus aureus protoplasts . Protoplasts could not be transformed with pGO200E (pGO200 on the staphylococcal replicon, pE194) or subclones containing DNA at one end of the tra fragment unless pGO1 or specific cloned tra DNA fragments were present in the recipient cell . However, once stabilized by sequences present on a second replicon, each tra fragment could be successfully introduced alone into other plasmid-free S . aureus recipients by conjugative mobilization or transduction . In this manner, two clones containing overlapping fragments comprising the entire 14.5-kb BglII fragment were shown to complement each other . The low-frequency transfer resulted in transconjugants containing one clone intact, deletions of that clone, and recombinants of the two clones . The resulting recombinant plasmid (pGO220), which regenerated the tra region intact on a single replicon, transferred at frequencies comparable to those of pGO1 . Thus, all the genes necessary and sufficient for conjugative transfer of pGO1 are contained within a 14.5-kb region of DNA.

J Hosp Infect, 1989 Feb, 13(2), 149 - 59
A method to evaluate the cleaning and disinfectant action of surface disinfectants; Walder M et al.; Surface disinfection tests, used to evaluate new disinfectants, do not take into account the effects of detergents or of the mechanical cleaning process . We describe methods which evaluate both the disinfection and cleaning effect of disinfectants on organic matter . When testing alcohols at high concentrations (greater than or equal to 70%) on blood spots contaminated with Staphylococcus aureus, we found that the organisms were trapped and fixed to the test surface, probably due to denaturation of the blood . This gave a low inactivating factor (IF), as well as a poor subjective cleaning effect (SC) . If serum was used instead of blood, we observed less pronounced trapping, resulting in a high IF although the SC was still poor . When broth was used, both IF and SC were satisfactory . With alcohols at a concentration of 42%, trapping was markedly reduced which improved the SC in blood contamination, with serum or broth contamination trapping did not occur . However, 42% ethanol lost its killing effect (i.e . low IF), whereas 42% isopropanol still demonstrated a high IF.

J Hosp Infect, 1989 Feb, 13(2), 133 - 41
Plasmid analysis of 219 methicillin-resistant Staphylococcus aureus strains with uncommon profiles isolated in Lisbon; Cristino JA et al.; During the years 1986 and 1987, 219 methicillin-resistant Staphylococcus aureus (MRSA) strains were isolated in a Lisbon hospital . Antimicrobial susceptibilities and genetic analysis showed that resistance to penicillin, methicillin, erythromycin (inducible phenotype), tetracycline, gentamicin, tobramycin, kanamycin, streptomycin, neomycin and trimethoprim were chromosomally encoded . Plasmid DNA was absent from 38.8% of the isolates . Constitutive erythromycin resistance was coded by three plasmids of c . 2.3 Md, c . 2.0 Md and c . 1.6 Md . Chloramphenicol resistance was mediated by two plasmids of c . 2.9 Md and c . 1.8 Md . Small cryptic plasmids of c . 1.65 Md, c . 1.2 Md and c . 1.0 Md were also detected . The majority of the strains revealed antigen 17, were lysed by phages 75, 89 and/or 85, and were either devoid of plasmid DNA, or possessed a c . 1.6 Md plasmid coding for constitutive erythromycin resistance or a c . 1.0 Md cryptic plasmid . These observations cannot rule out that the MRSA Lisbon isolates are a homogeneous group of strains that might have a common origin, and seem to be different from MRSA recently isolated in other countries.

J Hosp Infect, 1989 Feb, 13(2), 117 - 20
Nasal carriage of Staphylococcus aureus treated with topical mupirocin (pseudomonic acid) in a children's hospital; Frank U et al.; 2% mupirocin ointment applied intra-nasally for 5 days was assessed for elimination of nasal carriage of Staphylococcus aureus in 31 staff members in a children's hospital . Three volunteers failed to complete the trial because of side effects, i.e . buccal reddening and swelling, and unpleasant taste . During treatment staphylococcal nasal carriage was not found in any case; of the 24 post-treatment nasal swabs taken 4 days after treatment 22 were still negative . Re-colonization with S . aureus of different phage types occurred in the remaining two cases.

J Am Vet Med Assoc, 1989 Feb 1, 194(3), 389 - 91
Isolation of enterotoxigenic Escherichia coli from a foal with diarrhea; Holland RE et al.; Enterotoxigenic Escherichia coli was isolated from a 3-day-old foal with diarrhea . The isolate was distinguished from nonpathogenic E coli by determining the presence of pili and enterotoxin production . A standard slide agglutination test was performed, using pooled antisera that contained antibodies against K99 and F41 pilus antigens, K87 capsular antigen, and 0101 somatic antigen . Agglutination of the antisera occurred in the presence of the isolate . Piliation was verified by use of negative-contrast electron microscopy . Further, the isolate produced a heat-labile enterotoxin-like antigen that cross-reacted with a reagent containing formalin-treated, heat-killed Staphylococcus aureus (cowan 1 strain) bearing anti-cholera antibodies . On the basis of the aforementioned procedures and the absence of other identifiable enteric pathogens, we believe that E coli was responsible for causing diarrhea in the foal.

Anal Biochem, 1989 Feb 1, 176(2), 344 - 9
A new method of plasmid DNA preparation by sucrose-mediated detergent lysis from Escherichia coli (gram-negative) and Staphylococcus aureus (gram-positive); Saha B et al.; A simple and cheap method of plasmid DNA preparation from both gram-positive (Staphylococcus aureus) and gram-negative (Escherichia coli) organism is presented here . In this method, in place of the high-priced chemicals lysostaphin and lysozyme which are commonly used for removal of cell-wall during plasmid DNA preparation from gram-positive and gram-negative bacteria, respectively, only sucrose has been used . Firstly, bacteria is treated with Trizma (pH 8.0) containing 100% sucrose (hypertonic solution) . Due to this osmotic shock, protoplasm covered by the plasma membrane of bacteria possibly shrinks and becomes detached from the cell-wall . Osmotically sensitive cells thus formed, from gram-positive (S . aureus) and gram-negative (E . coli) bacteria, are finally lysed by the lysis mixture, containing brij 58 and sodium deoxycholate . The lysate is centrifuged at 15,000 rpm for 30 min to pellet the cell debris . The supernatant containing plasmid DNA is treated with either polyethylene glycol or isopropanol . The precipitate which contains plasmid DNA is dissolved in a buffer containing Tris, EDTA, NaCl, and sodium dodecyl sulfate (pH 8.0); thus protein is denatured and removed . Finally, RNA is removed by RNase treatment . The average yield of staphylococcal plasmid DNA as well as plasmid pBR322 from E . coli HB101 in 100% sucrose-treated preparations is greater than that of lysostaphin- and lysozyme-treated preparations . This method is applicable for both large-scale and small-scale preparations . The substrate activity for restriction enzyme, cloning, transforming ability, and electron microscopic profile of the plasmid DNA prepared by this method remains unaltered.

Anal Biochem, 1989 Feb 1, 176(2), 255 - 60
Strong-cation-exchange sulfoethyl aspartamide chromatography for peptide mapping of Staphylococcus aureus V8 protein digests; Crimmins DL et al.; In two recent reports (D . L . Crimmins, J . Gorka, R . S . Thoma, and B . D . Schwartz (1988) J . Chromatogr . 443, 63-71; A . J . Alpert and P . C . Andrews (1988) J . Chromatogr . 443, 85-96) a sulfoethyl aspartamide column was shown to efficiently analyze peptides less than 25 residues in length which differ in the number of nominal positive charges at pH 3.0 . In particular, the elution order for a series of distinct peptides ranging in nominal charge from +1 to +7 was found to be monotonic in nature indicating that separation was primarily via a cation-exchange mechanism . The present study employs this chromatographic system to isolate and characterize major fragments of proteolytic digests . Six commercially available proteins of known sequence (myoglobin, beta-casein, concanavalin A, carbonic anhydrase, lentil lectin, and enolase) were digested with Staphylococcus aureus V8 to generate peptide fragments . The resulting mixture was chromatographed on a sulfoethyl aspartamide column to isolate major fragments which were then subjected to amino acid analysis and N-terminal sequencing . With complete proteolysis (i.e., peptide fragments terminating in either an aspartic or a glutamic acid) separation of the fragments should result from the sum of histidine, lysine, and arginine residues contained in each fragment . Most of the peptide fragments eluted at the expected time on the sulfoethyl aspartamide column . Those fragments with anomalous behavior resulted from incomplete cleavage or cleavage at nonacidic residues or were greater than 35 residues in length . Each proteolytic digest was also analyzed by standard reverse-phase C4 chromatography to compare the peptide maps for these two distinct chromatographic modes.

J Trop Pediatr, 1989 Feb, 35(1), 6 - 9
Phagocyte metabolic functions in iron deficiency anaemia of Indian children; Hasan SM et al.; Nitroblue tetrazolium test (NBT) and bactericidal activity of polymorphonuclear leucocytes was studied in 40 patients with iron deficiency anaemia (aged 0-12 years) . NBT test had a significant correlation with serum iron (P less than 0.001) in all cases of iron deficiency anemia . Haemoglobin levels less than 4 g/dl also correlated significantly with NBT score . Bactericidal activity with opsonin-coated Staphylococcus aureus was considerably decreased in severe anaemia, but no significant correlation was found with serum iron concentration (P less than 0.1) . The phagocytic and bactericidal activity of polymorphonuclear leucocytes is dependent upon serum iron levels and is considerably reduced in iron deficient state in anaemic children.

APMIS, 1989 Feb, 97(2), 175 - 80
Hemagglutination by Staphylococcus aureus . Studies on strains isolated from bovine mastitis; Lindahl M et al.; Staphylococcus aureus strains isolated from bovine mastitis exhibited hemagglutinating activity when cultivated on nutrient-poor, solid medium . Out of 56 strains, 41 (73%) hemagglutinated . S . aureus agglutinated sheep erythrocytes to a greater extent than it agglutinated either human, bovine or equine erythrocytes . Hemagglutination was more conspicuous with erythrocytes from certain sheep individuals, indicating blood-group specificity . Receptors on sheep erythrocytes were apparently of carbohydrate nature since periodate-treated erythrocytes were not agglutinated . Hemagglutinating properties were associated with the ability to adhere to human buccal cells . Crude staphylococcal extracts were able to partially inhibit bacterial adherence to epithelial cells.

J Bacteriol, 1989 Feb, 171(2), 874 - 9
Involvement of multiple genetic determinants in high-level methicillin resistance in Staphylococcus aureus; Murakami K et al.; A methicillin-susceptible, novobiocin-resistant strain of Staphylococcus aureus (RN2677; methicillin MIC, 0.8 micrograms/ml) was transformed with DNA prepared from highly and homogeneously methicillin-resistant S . aureus strains (methicillin MIC, greater than or equal to 400 micrograms/ml) or from heterogeneous strains in which the majority of cells had a low level of resistance (methicillin MIC, 6.3 micrograms/ml) . All methicillin-resistant transformants showed low and heterogeneous resistance (methicillin MIC, 3.1 micrograms/ml) irrespective of the resistance level of DNA donors . All transformants examined produced normal amounts of the low-affinity penicillin-binding protein (PBP) 2a, and methicillin resistance and the capacity to produce PBP 2a showed the same degree of genetic linkage to the novobiocin resistance marker with both homogeneous and heterogeneous DNA donors . Next, we isolated a methicillin-susceptible mutant from a highly and homogeneously resistant strain which had a Tn551 insertion near or within the PBP 2a gene and thus did not produce PBP 2a . With this mutant used as the recipient, genetic transformation of the methicillin resistance gene was repeated with DNA isolated either from highly and homogeneously resistant strains or from heterogeneous (low-resistance) strains . All transformants obtained expressed high and homogeneous resistance and produced PBP 2a irrespective of the resistance level of the DNA donors . Our findings suggest that (i) the methicillin resistance locus is identical to the structural gene for PBP 2a, (ii) although the ability to produce PBP 2a is essential for resistance, the MICs for the majority of cells are not related to the cellular concentration of PBP 2a, and (iii) high MICs and homogeneous expression of resistance require the products of other distinct genetic elements as well.

J Pak Med Assoc, 1989 Feb, 39(2), 35 - 8
The carrier state: methicillin-resistant Staphylococcus aureus . A hospital study "screening of hospital personnel" for nasal carriage of Staph aureus; Ashiq B; Methicillin resistant Staph Aureus (MRSA) were studied in a 300 bedded Central Government Hospital Rawalpindi, in which 291 staff members were screened by nasal swabbing . Of 125 cases carrying staph aureus 5 (1.78%) were methicillin resistant . They were treated with Bacitracin ointment to be applied to interior nares four times a day for one week . Hexachlorophane baths daily, chlorhexidine shampoo once daily for a week, and were taken off duty from wards for one day.

Eur J Clin Microbiol Infect Dis, 1989 Feb, 8(2), 153 - 6
Comparative evaluation of a latex test for the identification of Staphylococcus aureus; Stevens M et al.; A rapid latex agglutination test, Staphaurex, was tested for its ability to identify Staphylococcus aureus using 72 reference strains and 785 clinical isolates of the family Micrococcaceae . All reference strains of Staphylococcus aureus were Staphaurex-positive . Non-Staphylococcus aureus reference strains were negative . Using clinical strains, the results of the Staphaurex test were compared with the results of other tests commonly used to identify Staphylococcus aureus . A total of 393 clinical isolates were classified as Staphylococcus aureus . The Staphaurex, slide coagulase, tube coagulase/human plasma and tube coagulase/rabbit plasma tests correctly identified 98%, 93.6%, 93.6% and 97.5% of the Staphylococcus aureus strains, respectively . The performance of the Staphaurex test, in terms of sensitivity and specificity, was significantly better than the slide coagulase test . It was as sensitive and almost as specific as the tube coagulase rabbit test and more sensitive than the tube coagulase human test.

J Interferon Res, 1989 Feb, 9(1), 143 - 51
Recombinant bovine interferon-gamma as an immunomodulator in dexamethasone-treated and nontreated cattle; Roth JA et al.; Three dosages (0.1, 0.5, and 2.5 mg/animal, subcutaneously), of recombinant bovine interferon-gamma (rBoIFN-gamma) were evaluated for their in vivo influence on neutrophil function and lymphocyte blastogenesis in cattle . The optimal of the three dosages tested (0.5 mg/animal or 1.1 x 10(6) U/animal) was then evaluated for its influence on neutrophils and lymphocytes in both normal and dexamethasone-treated cattle . One animal, which received 2.5 mg of rBoIFN-gamma, died by 24 h after administration due to acute diffuse interstitial pneumonia with interlobular edema and emphysema . The two highest dosages used caused fever at 24 h and the highest dosage caused a decrease in lymphocyte blastogenesis at 24 h after administration . The influence of rBoIFN-gamma on neutrophil function was dose dependent and depended on the baseline values for neutrophil function . Random migration by neutrophils was consistently inhibited in animals that received 0.5 mg or more of rBoIFN-gamma . Staphylococcus aureus ingestion and antibody-dependent cell-mediated cytotoxicity by neutrophils was enhanced by rBoIFN-gamma treatment in both dexamethasone-treated cattle and in nondexamethasone-treated cattle, which had relatively low values for these parameters before treatment . Iodination by neutrophils was also enhanced by rBoIFN-gamma when either a suboptimal concentration of neutrophil stimulant was used or when the cattle were treated with dexamethasone . In summary, the rBoIFN-gamma had greater immunomodulator activity in immunosuppressed than in normal cattle . The in vivo influence of rBoIFN-gamma therefore depends on the physiologic status of the animal.

J Interferon Res, 1989 Feb, 9(1), 115 - 24
Products of stimulated monocytes enhance the activity of interferon-gamma; Gerrard TL et al.; We have investigated the interaction of interferon-gamma (IFN-gamma) with monocytes or products of stimulated monocytes . We have shown that IFN-gamma does not stimulate IFN-alpha production in monocytes . Staphylococcus aureus (SAC) but not lipopolysaccharide (LPS) induced IFN-alpha secretion by monocytes . However, it was observed that supernatants of monocytes stimulated with IFN-gamma in combination with either LPS or SAC had higher levels of antiviral activity than supernatants of monocytes stimulated only with IFN-gamma . Moreover, the degree of enhancement of antiviral activity was dependent on the dose of either LPS or SAC used to stimulate the monocytes . Supernatants of monocytes stimulated with LPS or SAC enhanced the antiviral activity of IFN-gamma but not IFN-alpha . Thus, LPS- or SAC-stimulated monocytes produced a factor(s) that augmented the biological activity of IFN-gamma . To identify the factor within stimulated monocyte supernatants that was responsible for this enhancement, several monokines were added to IFN-gamma . Tumor necrosis factor (TNF) significantly increased the antiviral activity of IFN-gamma, although TNF by itself had no antiviral activity . Interleukin 1 (IL-1) or granulocyte-monocyte colony-stimulating factor (GM-CSF) did not enhance the activity of IFN-gamma . Our data indicate that the interaction between IFN-gamma and monocytes is bidirectional . Not only can IFN-gamma activate monocytes, but products of stimulated monocytes also enhance the biological activities of IFN-gamma.

J Leukoc Biol, 1989 Feb, 45(2), 114 - 20
Staphylococcus aureus tetrapeptide with high chemotactic potency and efficacy for human leukocytes; Rot A et al.; A chemotactic tetrapeptide from culture fluids of Staphylococcus aureus was purified to homogeneity by reverse-phase high-pressure liquid chromatography . The peptide comprises equimolar methionine, leucine, isoleucine, and phenylalanine . It inhibited binding of fluoresceinated fMet-Leu-Phe-Lys to human monocytes, which showed that it interacted with the formyl-methionyl peptide receptor and suggested that it was a formyl-methionyl peptide . Based on a comparison of dose-response curves for inhibition of fluoresceinated fMet-Leu-Phe-Lys binding, the relative affinity of the peptide for the receptor was comparable to that of fMet-Leu-Phe-Lys . At optimal concentrations, chemotactic efficacy (percentage of monocytes migrating to the attractant) was 53 +/- 4%, in contrast to 36 +/- 3% for the reference attractant fMet-Leu-Phe . Since approximately 60% of human monocytes have formyl-peptide receptors, the bacterial peptide is capable of attracting all receptor-bearing monocytes.

J Clin Microbiol, 1989 Feb, 27(2), 335 - 6
Methicillin-resistant strains of Staphylococcus aureus resistant to quinolones; Schaefler S; Since January 1988, Staphylococcus aureus strains with a high level of quinolone resistance have been isolated at 17 hospitals and 15 nursing homes in New York City . The majority of these strains were methicillin resistant . The bacteriophage types and susceptibility to other antibiotics were similar to those of quinolone-susceptible strains isolated at the same hospitals.

J Virol, 1989 Feb, 63(2), 481 - 92
Transduction of the cellular src gene and 3' adjacent sequences in avian sarcoma virus PR2257; Geryk J et al.; When injected into chickens, a transformation-defective mutant of the Prague C strain of Rous sarcoma virus induced tumors at low incidence and after a long latency . One such tumor released a replication-defective virus designated PR2257 . We molecularly cloned and sequenced the proviral DNA from quail fibroblasts transformed by PR2257 . Comparison of PR2257 sequence with that of Prague C, cellular src, and 3' adjacent cellular DNA showed that the spliced version of the c-src gene and about 950 base pairs (bp) of 3'-flanking cellular DNA were transduced into PR2257 . This transduction eliminated nearly all replicative genes, since the gag gene splice donor site was linked to the splice acceptor site of the src gene and, on the 3' side, recombination occurred in the end of env gene . Insertion of two extra cytosines 23 bp before and 19 bp after the c-src stop codon resulted in an extension of the coding portion up to 587 amino acids, divergence of sequences after Pro-525 and replacement of Tyr-527 by a valine residue . In addition, it appears that the 5' and 3' untranslated regions of PR2257 result from multiple recombinations between exogenous and endogenous virus genomes . Limited digestion of p66src encoded by PR2257 with Staphylococcus aureus V8 protease yielded a V2 peptide (C-terminal moiety) with an apparent molecular mass of 31 kilodaltons, consistent with the 5.7-kilodalton increase expected from the DNA sequence . The structure of PR2257 suggests that the first step in the capture of c-src gene by avian lymphomatosis viruses is the trans splicing of the viral leader mRNA to exon 1 of c-src.

Cell Immunol, 1989 Feb, 118(2), 368 - 81
Regulatory role of CD19 molecules in B-cell activation and differentiation; de Rie MA et al.; Cluster of differentiation ({CD}) 19 antigens are B-cell-specific molecules expressed on virtually all human cells of the B-lymphocyte lineage except plasma cells . We produced a new anti-CD19 monoclonal antibody (McAb), CLB-CD19, that was used to study the role of CD19 molecules in B-cell activation . Anti-CD19 McAb induced mobilization of free intracellular calcium ({Ca2+}i) in Daudi cells, but not in normal spleen or tonsillar B cells, for which crosslinking with a second anti-mouse Ig antibody was not required . Anti-CD19 McAb inhibited B-cell proliferation induced by anti-IgM coupled to Sepharose beads . This inhibitory effect was overcome by the addition of nonmitogenic concentrations of phorbol myristate acetate . Anti-CD19 McAb did not interfere with Staphylococcus aureus- or B-cell growth factor-induced B-cell proliferation . Anti-CD19 McAb inhibited T-cell-dependent polyclonal B-cell differentiation in pokeweed mitogen-, interleukin 2-, or anti-CD3-driven culture systems . Delayed addition studies showed that once differentiation of B cells was induced, CD19 molecules lost their regulating function . Taken together, our results indicate that CD19 molecules play a regulatory role in B-cell proliferation and differentiation.

FEBS Lett, 1989 Jan 30, 243(2), 209 - 12
Complete amino acid sequence of the sarcoplasmic calcium-binding protein (SCP-I) from crayfish (Astacus leptodactylus); Jauregui-Adell J et al.; The complete amino acid sequence of the alpha chain of the dimeric sarcoplasmic Ca2+-binding protein (SCP-I = alpha 2) from crayfish (Astacus leptodactylus) has been determined by partial automatic sequencing of the peptides derived from tryptic digests of the protein after citraconylation or treatment with 1,2-cyclohexanedione . Overlapping peptides were obtained by cleavage with o-iodosobenzoic acid, or digestion with Staphylococcus aureus protease, thermolysin and pepsin . The acetylated N-terminus was identified by fast atom bombardment mass spectrometry . The monomeric protein contains 192 amino acids and has an Mr of 21,643 . The sequence shows the presence of three calcium-binding sites and perhaps of two others that may be degenerated.

Med Clin (Barc), 1989 Jan 21, 92(2), 47 - 51
{Prospective study of 75 episodes of sepsis in hemodialysed patients}; Martinez JA et al.; During a 38-month period the bacteremias developing in patients on hemodialysis from three centers of the Barcelona area were evaluated to assess their incidence, characteristics, and response to therapy . In the overall 13376 months of hemodialysis of the study, 75 episodes of bacteremia were detected in 64 patients; this amounts to an incidence of 5.6 episodes per 1000 hemodialysis months . The most common sources of becteremia were intravenous catheters (44%), which were mainly used as temporary vascular access, followed by the definitive vascular access (26%), the genitourinary system (10%), and the lung (6%) . Twenty-seven episodes of bacteremia developed in 24 patients in whom dialysis had been started in the two previous months (11% of the overall number of new patients), and, in them, 77% originated in an intravenous catheter, while this was the origin of bacteremia in only 23% of the remaining patients . 72% of bacteremias were caused by gram positive organisms, particularly Staphylococcus aureus and Staphylococcus epidermidis (60%), followed, in frequency order, by aerobic gram negative bacilli (25%), particularly Escherichia coli and Pseudomonas . Stpahylococci were significantly associated with the vascular access, either if this was a catheter or not (81% of instances), while gram negative bacilli were associated with sources different from the vascular access (48% of instances) . Severe complications included 2 cases of aortic valve endocarditis, one hemorrhagic shock caused by rupture of an infected vascular access, and one suppurative phlebitis associated with a hemodialysis catheter . Staphylococcal sepsis was randomly treated with vancomycin or vancomycin plus gentamicin, without differences in the results.(ABSTRACT TRUNCATED AT 250 WORDS)

Biochem Biophys Res Commun, 1989 Jan 16, 158(1), 319 - 25
alpha B subunit of lens-specific protein alpha-crystallin is present in other ocular and non-ocular tissues; Bhat SP et al.; alpha-Crystallin, a tissue specific structural protein of the ocular lens, is known to be composed of two subunits, alpha A and alpha B . By using a specific antibody in an immunoblotting procedure we have found that one of the subunits, alpha B is present in a number of non-lenticular tissues including the retina, heart, skeletal muscle, skin, brain, spinal cord and lungs . Interestingly, in the rat, this protein is present in significantly higher concentrations in adult than in fetal tissues and, with the exception of the lens, fetal and adult heart has the highest concentration among the tissues examined . That the protein in question is, in fact, alpha B, was confirmed a) by the remarkable similarity of Staphylococcus aureus protease peptide maps of the protein in the heart and purified alpha-crystallin and b) by the sequence analysis of a rat heart cDNA clone identified by the alpha B antibody . Based on these observations we conclude that while alpha A has a tissue-specific role, alpha B is a polypeptide of independent function not restricted to the ocular lens.

Med J Aust, 1989 Jan 16, 150(2), 65, 69 - 72
A national survey of antimicrobial resistance in Staphylococcus aureus in Australian teaching hospitals; Turnidge J et al.; An extensive survey of the in-vitro susceptibility of clinical isolates of Staphylococcus aureus to 18 antimicrobial agents was conducted over three separate periods during 1986-1987 in 14 teaching hospitals in major Australian cities . The survey aimed to document the prevalence of resistance to a wide variety of drugs that are important as antistaphylococcal agents or as epidemiological markers . More than 7500 isolates were examined . Nationally, the prevalence of resistance was 85.3% to penicillin G, 14.4% to methicillin, 14.0% to amoxycillin/clavulanate, 9% to cephalothin, 5.4% to cephamandole, 9.9% to cefotaxime, 25.0% to erythromycin, 11.2% to clindamycin, 21.7% to tetracycline, 13.0% to gentamicin, 1.9% to amikacin, 5.8% to chloramphenicol, 18.3% to trimethoprim, 0.6% to rifampicin, 3.0% to fusidic acid and 1.2% to novobiocin . For none of the strains was resistance to vancomycin confirmed by minimal-inhibitory-concentration determination . A high proportion of the resistances was harboured in methicillin-resistant Staph . aureus, except for resistance to penicillin G, erythromycin and tetracycline . The prevalence of methicillin resistance varied widely among the states: 25.2% in Queensland, 23.5% in Victoria, 12.6% in New South Wales/the Australian Capital Territory, 11.3% in South Australia and 0.4% in Western Australia . Isolates from blood cultures were slightly-more susceptible to antimicrobial agents than were isolates from other body sites . Six common profiles of resistance to penicillin G, methicillin, erythromycin, clindamycin and tetracycline accounted for more than 95% of the isolates that were tested against all five agents . Vancomycin remains the most important antistaphylococcal drug in areas where resistance to methicillin is common.

J Biol Chem, 1989 Jan 15, 264(2), 804 - 9
High level expression in Escherichia coli of the DNA-binding domain of the glucocorticoid receptor in a functional form utilizing domain-specific cleavage of a fusion protein; Dahlman K et al.; A fragment comprising the DNA-binding domain of the human glucocorticoid receptor has been expressed in a functional form in Escherichia coli as a fusion protein with protein A from Staphylococcus aureus . The DNA-binding domain was purified to apparent homogeneity by affinity chromatography on IgG-Sepharose and DNA-cellulose, a purification scheme which does not involve denaturation of the protein at any step . The DNA-binding domain was separated from the protein A part of the fusion protein by domain-specific enzymatic cleavage with chymotrypsin while immobilized on IgG-Sepharose . The recombinant protein has been characterized by amino acid analysis, NH2- and COOH-terminal sequence analysis, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and reactivity to iodoacetate and was found to correspond to the primary structure derived from the cDNA sequence . DNase I footprinting showed that the purified recombinant protein bound to the same DNA sequences on the mouse mammary tumor virus long terminal repeat as glucocorticoid receptor purified from rat liver does . About 10 times more recombinant protein, on a molar basis, was needed to obtain the same level of protection . However, the protection of the three different footprints (1.3, 1.4, and 1.5') by the recombinant protein differed greatly from that of the natural receptor, with virtually no protection of footprint 1.4 . This indicates cooperative binding of the natural receptor to adjacent footprints, dependent on other regions of the receptor than the DNA-binding domain.

J Biol Chem, 1989 Jan 5, 264(1), 40 - 4
Selective binding of L-thyroxine by myosin light chain kinase; Hagiwara M et al.; L-Thyroxine selectively inhibited Ca2+-calmodulin-activated myosin light chain kinases (MLC kinase) purified from rabbit skeletal muscle, chicken gizzard smooth muscle, bovine thyroid gland, and human platelet with similar Ki values (Ki = 2.5 microM) . A detailed analysis of L-thyroxine inhibition of smooth muscle myosin light chain kinase activation was undertaken in order to determine the effect of L-thyroxine on the stoichiometries of Ca2+, calmodulin, and the enzyme in the activation process . The kinetic data indicated that L-thyroxine does not interact with calmodulin but, instead, through direct association with the enzyme, inhibits the binding of the Ca2+-calmodulin complex to MLC kinase . L-{125I}Thyroxine gel overlay revealed that the 95-kDa fragment of chicken gizzard MLC kinase digested by chymotrypsin and all the fragments of 110, 94, 70, and 43 kDa produced by Staphylococcus aureus V8 protease digestion which contain the calmodulin binding domain retain L-{125I}thyroxine binding activity, whereas smaller peptides were not radioactive . Since MLC kinase is phosphorylated by cAMP-dependent protein kinase (2 mol of phosphate/mol of MLC kinase), the effect of L-thyroxine on the phosphorylation of MLC kinase also was examined . L-Thyroxine binding did not inhibit the phosphorylation of MLC kinase and, moreover, reversed the inhibition of phosphorylation obtained with the calmodulin-enzyme complex . These observations support the suggestion that L-thyroxine binds at or near the calmodulin-binding site of MLC kinase . L-Thyroxine may serve as a different type of pharmacological tool for elucidating the biological significance of MLC kinase-mediated reactions.

J Biol Chem, 1989 Jan 5, 264(1), 54 - 60
Enzymatic methylation of L-isoaspartyl residues derived from aspartyl residues in affinity-purified calmodulin . The role of conformational flexibility in spontaneous isoaspartyl formation; Ota IM et al.; We have investigated the formation of D-aspartyl and L-isoaspartyl (beta-aspartyl) residues and their subsequent methylation in bovine brain calmodulin by the type II protein carboxyl methyltransferase . Based on the results of studies with unstructured peptides and denatured proteins, it has been proposed that the major sites of carboxyl methylation in calmodulin are at L-isoaspartyl residues that originate from two Asn-Gly sequences . To test this hypothesis, we directly identified the sites of methylation in affinity-purified preparations of calmodulin by peptide mapping using the proteases trypsin, endoproteinase Lys-C, clostripain, chymotrypsin, and Staphylococcus aureus V8 protease . We found, however, that the major high-affinity sites of methylation originate from aspartyl residues at position 2 and at positions 78 and/or 80 . The methylatable residue in the first case was shown to be L-isoaspartate by comparison of the properties of a synthetic peptide corresponding to the N-terminal 13 residues substituted with an L-iso-Asp residue at position 2 . The second methylatable residue, probably derived from Asp78, also appears to be an L-isoaspartyl residue . These sites appear to be readily accessible to the methyltransferase and are present in relatively flexible regions of calmodulin that may allow the spontaneous degradation reactions to occur that generate L-isoaspartyl residues via succinimide intermediates . Interestingly, the four calcium binding regions, each containing 3-4 aspartyl and asparaginyl residues (including the two Asn-Gly sequences), do not appear to contribute to the high-affinity methyl acceptor sites, even when calcium is removed prior to the methylation reaction . We propose that methylatable residues do not form at these sites because of the inflexibility of these regions when calcium is bound.

Med J Aust, 1989 Jan 2, 150(1), 25 - 8
Bacterial infections among patients with diabetes in Papua New Guinea; Patel MS; This study reports the morbidity that resulted from bacterial infections in Melanesian patients with non-insulin-dependent diabetes who attended the Port Moresby General Hospital, Papua New Guinea, between January 1, 1982 and June 30, 1984 . Fifty-three of 160 patients with diabetes experienced 66 episodes of infection, 48 of which required inpatient hospital treatment . The average length of stay in hospital was 37.6 days per episode of infection . Of 88 patients who were newly-diagnosed as diabetic during this period, 30 patients initially had presented with a bacterial infection . The lower limb was the site that was infected most frequently, and Staphylococcus aureus and Klebsiella pneumoniae were the usual causative organisms . Eleven patients had bacterial gangrene of the foot; two of these patients were less than 23 years of age, and five patients were not known to have had diabetes previously . Five patients were suffering from pulmonary tuberculosis; the annual incidence of tuberculosis in this study group (12.5 cases/1000 patients) was about 11-times higher than that which has been reported for the general population . Thirteen patients with diabetes died in hospital during the study period . Infection was the cause of death in nine patients and three of these patients were less than 25 years of age . The morbidity of infection can be controlled if diabetes is sought more frequently in patients with infections, and if glycaemia can be controlled . This will have to be achieved through existing primary health-care structures, as resources for diabetes-specific preventive programmes in developing countries will be limited.

Rev Infect Dis, 1989 Jan-Feb, 11 Suppl 1, S90 - 5
Distribution and expression of toxic shock syndrome toxin 1 gene among Staphylococcus aureus isolates of toxic shock syndrome and non-toxic shock syndrome origin; Bonventre PF et al.; Toxic shock syndrome toxin 1 (TSST-1), plays a significant role in the pathogenesis of TSS . TSST-1 production is subject to physiologic and environmental constraints . Thus, DNA probes that detect the chromosomal gene encoding the toxin are of value diagnostically, epidemiologically, and for studies of gene expression . Several synthetic oligonucleotide probes complementary to two regions of the TSST-1 gene were used to ascertain the presence of this gene in the chromosomal DNA of 261 strains of S . aureus from various TSS-related and non-TSS-related sources . Isolates were from clinically confirmed menstrual and nonmenstrual cases of TSS and from healthy vaginal carriers of S . aureus . Other strains tested included clinical non-TSS isolates and food poisoning-associated staphylococcal isolates . Detection of the TSST-1 gene by the labeled gene probes correlated in all but two cases with production of TSST-1 . Ten Centers for Disease Control (CDC) strains that were isolated from TSS patients and did not produce TSST-1 were also examined, as were several strains of Staphylococcus epidermidis isolated from patients with suspected TSS . Neither group of strains possessed the TSST-1 gene . Finally, a 7-kilobase DNA restriction fragment of S . aureus containing the entire TSST-1 gene was transformed into Escherichia coli strains HB101 and DH5 alpha via a plasmid vector.

Rev Infect Dis, 1989 Jan-Feb, 11 Suppl 1, S313 - 5
Direct effects of purified staphylococcal toxic shock syndrome toxin 1 on myocardial function of isolated rabbit atria; Olson RD et al.; Toxic shock syndrome is associated with reversible cardiomyopathy . The cardiac dysfunction may be mediated by toxic shock syndrome toxin 1 (TSST-1), an exotoxin generated by Staphylococcus aureus . However, the effects of purified TSST-1 on cardiac function are unknown . In a study of the toxin's effect on myocardial function, TSST-1 (200 ng/mL) was added to muscle baths containing isolated rabbit atria . TSST-1 caused time-dependent inhibition of developed force (systolic function) but did not alter resting force (an index of muscle stiffness or cardiac compliance) . These data show that TSST-1 can directly inhibit myocardial function, but the significance of this effect in patients with toxic shock syndrome remains to be determined.

Rev Infect Dis, 1989 Jan-Feb, 11 Suppl 1, S231 - 6; discussion S236-7
Response of various animal species to experimental infection with different strains of Staphylococcus aureus; Kohrman KA et al.; An experimental infection program was conducted in rabbits, pigs, and baboons with toxic shock syndrome (TSS)-associated and non-TSS-associated strains of Staphylococcus aureus to produce an animal model for TSS . TSS-associated strains of S . aureus--whether positive or negative for TSS toxin 1 (TSST-1)--could not be distinguished from non-TSS-associated strains of S . aureus by means of the rabbit whiffle-ball infection model; therefore, limited pilot infection studies were conducted in pigs and baboons . Experimental conditions were optimized in both the pig and the baboon studies to maximize the chance of producing TSS . Pigs infected with TSS-associated S . aureus strain CDC-11 developed some of the clinical signs observed in TSS (fever, hypotension, diarrhea, and vomiting) . However, no changes were detected in clinical chemistry or hematology . Baboons infected with S . aureus strain CDC-11 showed only minimal signs of illness, i.e., lethargy, decreased food intake, and loose stools . TSS was not produced in pigs or baboons, even under optimal exposure conditions.

Rev Infect Dis, 1989 Jan-Feb, 11 Suppl 1, S214 - 7; discussion S217-8
Passive protection of rabbits infected with toxic shock syndrome-associated strains of Staphylococcus aureus by monoclonal antibody to toxic shock syndrome toxin 1; Scott DF et al.; The effectiveness of passive immunization was assessed in an infection model of toxic shock syndrome (TSS) in which monoclonal antibody to TSS toxin 1 (TSST-1) was administered intravenously to rabbits . Previously implanted infection chambers were inoculated with Staphylococcus aureus strains RN4710 and D4508 . The former strain carries the TSST-1 gene on plasmid pRN6201; the latter is a TSST-1-negative clinical isolate obtained from a patient with nonmenstrual TSS . Purified monoclonal antibody, MAb 8-5-7 (IgG), was administered in two doses of approximately 1.25 mg each 24 hours before and 24 hours after infection . MAb 8-5-7 provided complete protection against both the TSS-like syndrome and the mortality that occurred in unprotected rabbits infected with strain RN4710 but did not provide complete protection in rabbits infected with strain D4508; three of the five rabbits either displayed signs of illness or died despite treatment . Western-blot analyses of the extracellular proteins produced by strains RN4710 and D4508 that used MAb 8-5-7 as a probe revealed that only TSST-1 produced by RN4710 reacted with the antibody . Thus, if MAb 8-5-7 partially protected animals against infections with strain D4508, the protection appears to have been nonspecific.

Rev Infect Dis, 1989 Jan-Feb, 11 Suppl 1, S197 - 202
Effect of tampon wraps on production of toxic shock syndrome toxin 1; Robbins RN et al.; The influence of tampon wraps on the production and distribution of toxic shock syndrome toxin 1 (TSST-1) was investigated with use of a disk-membrane-agar method . Filter membranes (45 micron) overlaying agar medium in 50-mm petri dishes were spread-inoculated with a TSST-1-producing strain of Staphylococcus aureus and covered with 0.5 mL of whole citrated rabbit blood . Disks of tampon core materials with and without wraps were placed on the inoculated membranes and incubated at 37 degrees C in 5% CO2 in air . Eight wraps with each of 14 tampon core materials were tested to determine the amount of toxin in the agar layer, a quantity considered an indicator of the relative amounts of toxin available for absorption in vivo . The average level of toxin in the agar layer with any particular wrap compared with that in the agar layer with no wrap was higher with five wraps, lower with two wraps, and essentially the same with one wrap . The lower levels of toxin were obtained with one wrap that had a starch binder and with one that contained the surfactant Standamul 1414-E . Standamul 1414-E has been reported to be an inhibitor of growth of and production of TSST-1 by S . aureus.

Rev Infect Dis, 1989 Jan-Feb, 11 Suppl 1, S188 - 95; discussion S195-6
Growth of Staphylococcus aureus and synthesis of toxic shock syndrome toxin 1 in different in vitro systems; Kirkland JJ et al.; Various complex and synthetic bacterial growth media have been used to study the growth of Staphylococcus aureus and the production of toxic shock syndrome toxin 1 (TSST-1) under certain in vitro culture conditions . Because of the biochemical and nutritional differences between these media and human menses, a program was designed to determine the growth and metabolism of S . aureus under conditions that more closely approximate in vivo conditions . Human menses, an artificial menses developed to match human menses, whole human blood, and complex bacterial culture media (with and without added whole human blood) were tested for the ability to support the growth of S . aureus and the production of TSST-1 in vitro . In addition, the impact of other organisms, commonly isolated from the human vagina, on the growth of S . aureus and the production of TSST-1 was evaluated . Results show that the environmental conditions provided by human menses are more closely approximated by the artificial menses and that neither commercial bacterial growth media alone nor complex media plus 25% or 50% human blood provide a menses-like environment for the growth of S . aureus and the production of TSST-1 . Furthermore, the addition of a second organism to the S . aureus culture resulted in significant suppression of TSST-1 production, even when in vitro conditions were optimized for the production of TSST-1.

Rev Infect Dis, 1989 Jan-Feb, 11 Suppl 1, S182 - 6; discussion S186-7
Ecology of toxic shock syndrome: amplification of toxic shock syndrome toxin 1 by materials of medical interest; Tierno PM Jr et al.; Historically, the literature suggests that staphylococcal exoproteins, including enterotoxins, are stimulated by various physicochemical ecologic factors, many of which have been shown to stimulate production of toxic shock syndrome toxin 1 (TSST-1) . The propensity of different fibers and other substances to amplify TSST-1 production in toxic shock syndrome-associated strains of Staphylococcus aureus, as well as a comparative analysis of the underlying mechanisms of TSST-1 production, are reported . Two hundred twenty intravaginal devices or other products and materials and 60 experimental controls were examined for their propensity to induce TSST-1 production . Certain materials are superior to unaltered cotton in providing a more absorbent fiber--nutrients are efficiently drawn in, concentrating protein between fibers, and thereby creating an ideal physicochemical environment for the amplification of TSST-1 and other toxins . The greatest stimulation of TSST-1 was observed with (in decreasing order): polyester and carboxymethyl cellulose, polyacrylates, viscose rayon, gelatin foam, polyurethane, and cotton . No toxin was found with nasal tampons (polymer of polyvinyl acetal) or with vaginal cups (an elastomeric polymer) . Results are discussed in terms of specific ecologic parameters from historical as well as recent perspectives.

Rev Infect Dis, 1989 Jan-Feb, 11 Suppl 1, S157 - 66
Effect of magnesium on in vitro production of toxic shock syndrome toxin 1; James JF et al.; Batch and chemostat culture of Staphylococcus aureus strain S411 was conducted in an investigation of the role of Mg++ in the control of production of toxic shock syndrome toxin 1 (TSST-1) . Under both growth conditions, Mg++ influenced bacterial growth, and TSST-1 production was correlated with bacterial growth . The specific activity of TSST-1 (ng/mg yield, ng/mg total protein) increased with increasing concentrations of Mg++ and was maximal at physiologic levels of Mg++ . No production of TSST-1 was observed under anaerobic conditions . In chemostat cultures in which valine nutrient limitation was used with various levels of tryptophan in the chemically defined medium, tryptophan concentration controlled the production of TSST-1 by strain S411, regardless of the concentration of Mg++.

Rev Infect Dis, 1989 Jan-Feb, 11 Suppl 1, S151 - 6
Production of toxic shock syndrome toxin 1 by Staphylococcus aureus under aerobic and anaerobic conditions and the effect of magnesium ion limitation; Taylor D et al.; A toxic shock syndrome isolate of Staphylococcus aureus was grown in a chemostat in a defined synthetic medium of six amino acids, glucose, two vitamins, and salts . Steady states were achieved under limiting and replete magnesium ion conditions as well as with and without oxygen . The biomass and the amounts of toxic shock syndrome toxin 1, acid phosphatase, proteinase, hyaluronate lyase, and total exoprotein were estimated at each condition . Under magnesium limitation all variables, measured as specific production rates, were reduced--apart from proteinase--compared with the magnesium-replete condition . A similar pattern of results occurred with comparison of anaerobically and aerobically grown cells.

Rev Infect Dis, 1989 Jan-Feb, 11 Suppl 1, S145 - 49; discussion S149-50
Effect of trace metals on the synthesis of toxic shock syndrome toxin 1; Reeves MW; The effect of 10 trace metals on the production of toxic shock syndrome toxin 1 (TSST-1) was studied in a metal-deficient chemically defined medium (CDM) with toxic shock syndrome (TSS) and non-TSS strains of Staphylococcus aureus . When individual metals were tested, only Mg++ stimulated cell growth and TSST-1 production . TSST-1 yield was responsive to the concentration of Mg++ in the medium, with a maximal yield occurring at 0.1 mM . When other metals were studied in the presence of Mg++ with and without 1 mM ethylenediamine tetraacetic acid, it was found that Ca++, Co++, Cu++, Fe++, Mn++, molybdate-, Ni++, vanadate-, and Zn++ produced some stimulation of either cell growth or TSST-1 synthesis or both . The stimulating effect of Mg++ on TSST-1 synthesis was significantly enhanced by adding 0.3 mM Zn++ and 0.003 mM Fe++ . TSST-1 production by 11 strains of S . aureus in CDM with optimal concentrations of trace metals was two to seven times greater than that produced in brain-heart infusion broth.

Rev Infect Dis, 1989 Jan-Feb, 11 Suppl 1, S130 - 5; discussion S135-6
Identification of a new toxin from a strain of Staphylococcus aureus isolated from a patient with nonmenstrual toxic shock syndrome; Hall WW et al.; The extracellular products of a toxic shock syndrome toxin 1 (TSST-1)-negative isolate of Staphylococcus aureus (strain D4508) from a patient with nonmenstrual toxic shock syndrome were shown to possess toxic activity capable of producing shock and death when injected into rabbits . Fractionation of the extracellular products and the production of polyclonal antibodies to individual polypeptides identified the toxin as a single polypeptide of molecular weight 26,000 . Immunoblot analyses showed the toxin to be antigenically related yet distinct from enterotoxins A, B, C, D, and E and unrelated to TSST-1.

Eksp Onkol, 1989, 11(1), 73 - 4, 79
{Effect of staphylococcal enterotoxin A on the development of Lewis lung carcinoma in mice}; Shcheglovitova ON et al.; The effect of Staphylococcus aureus enterotoxin A (SEA) was studied for its effect on the development of the Lewis carcinoma in mice . It was shown that administration of SEA immediately after the appearance of the primary node in mice after transplantation of tumour cells led to insignificant inhibition of the node growth and a slight decrease of tumour metastasizing into the lungs . Inoculation of mice after the appearance of the primary node with 1/microgram of SEA 5 times a week significantly increased their survival rate . The lack of the marked effect of SEA appears to be associated with the disturbance of the immune interferon system functioning in tumour-bearing mice, since the production of serum interferon induced by SEA in mice with tumours was considerably lower than in the intact ones.

Res Vet Sci, 1989 Jan, 46(1), 1 - 4
Prevalence of staphylococcal species in four dairy herds; Watts JL et al.; The prevalence of staphylococcal species isolated from bovine mammary glands was determined in four dairy herds . Staphylococcus aureus and S hyicus were the predominant organisms isolated from cows in a herd with a bulk milk somatic cell count (SCC) greater than 900 X 10(3) . One herd with a bulk milk SCC of 565 X 10(3) had a high incidence of S aureus while the predominant coagulase-negative species were S epidermidis and S hyicus . S hyicus predominated in two herds with bulk milk SCC less than 200 X 10(3); prevalence of S aureus was low . The impact of herd management practices such as post-milking teat antisepsis on distribution of staphylococcal species is discussed.

Hautarzt, 1989 Jan, 40(1), 19 - 22
{Specific IgE to Staphylococcus aureus in patients with atopic dermatitis}; Kapp A et al.; It is a well-known feature of atopic dermatitis (AD) that the patient's skin is heavily colonized by Staphylococcus aureus . S . aureus-derived antigens may be important triggers of the immune response and may significantly contribute to the genesis of the cutaneous pathology of AD . Therefore, serum samples of 52 patients with AD, all of whom had signs of moderate to severe disease activity, were tested for antistaphylococcal IgE antibodies with RAST discs coupled to antigens derived from Wood 46 strain . Total IgE concentrations and specific IgE to nine different common allergens were also determined . Only 2 patients showed significant levels of specific IgE antibodies to S . aureus (RAST class greater than or equal to 2) . Both these patients were found to have high total IgE and significant levels of specific IgE to all nine common allergens tested . One of the patients had marked eosinophilia . We conclude that the presence of specific IgE to S . aureus is not correlated with the disease activity in AD . Specific antistaphylococcal IgE does not represent an important diagnostic feature in AD, but may be of importance for the detection of subgroups within patients affected by AD.

Acta Paediatr Scand, 1989 Jan, 78(1), 56 - 61
Staphylococcus aureus bacteraemia in children below the age of one year . A review of 407 cases; Espersen F et al.; The bacteriology, epidemiology, infection types and mortality were reviewed in 407 cases of Staphylococcus aureus bacteraemia in the period 1967-1984 in children less than one year old . The number of bacteraemia cases in this age group increased in the seventies, but only 4% of the bacteraemias seemed to be due to epidemic spreading . Neonates might be at higher risk for S . aureus bacteraemia than older children less than one year old, and nearly all infections in the neonatal period were hospital-acquired . The mortality was 24%, and did not correlate with type of infection, but in neonates it correlated with low birthweight.

Lab Anim Sci, 1989 Jan, 39(1), 51 - 5
Chlorine dioxide sterilization of implanted right atrial catheters in rabbits; Dennis MB Jr et al.; The disinfection of right atrial catheters in situ using chlorine dioxide was investigated . Catheters were implanted into rabbits (Oryctolagus cuniculi) and colonized by inoculation of Staphylococcus aureus, Sa-80, into the lumen . All of the catheters were colonized and the difference in numbers of bacteria recovered from animals destined for the control and disinfection groups was not significant . Animals were assigned randomly to the control or disinfection group . Treatment consisted of filling the catheter lumen of the disinfection group with chlorine dioxide and of the control group with sterile physiological saline daily for 15 minutes . In addition, both groups received systemic antimicrobial therapy . Cultures of blood withdrawn from the catheters and by venipuncture were negative for five of the nine control group animals after treatment for 5 days . Four control group catheters failed, after from 3 to 21 treatments, without ever achieving negative cultures . All nine animals in the disinfection group had negative cultures after treatment for 5 days . Subsequently, one animal from each group reverted to positive cultures . All nine control group catheters failed during the study, compared to only three disinfection group catheters (p less than 0.01) . At necropsy, culture of cardiac blood, thrombi and catheter tubing sections demonstrated colonization of six in the control group and only one in the disinfection group (p less than 0.05) . Rabbits tolerated the chlorine dioxide disinfection well and no adverse signs were noted.

Indian J Med Res, 1989 Jan, 89, 40 - 2
Phagocytic function of monocytes in murine model of Echinococcus granulosus of human origin; Wangoo A et al.; Phagocytic function of monocytes was studied in E . granulosus infected Swiss albino mice, with haemolysin coated sheep erythrocytes, Staphylococcus aureus, latex particles and- Echinococcus antigen coated latex particles . No significant difference in percentage phagocytic activity was observed in mice in the early stages of infection when compared to non-infected controls (P greater than 0.05) . However, in the later stages of infection, increase in the phagocytic activity was noticed . The activity was more prominent against latex particles coated with Echinococcus antigen (P less than 0.001) . Although the significance of these responses is not clear, this study showed an intact inflammatory response with certain degree of specificity.

J Med Microbiol, 1989 Jan, 28(1), 25 - 32
Characterisation of methicillin-resistant isolates of Staphylococcus aureus by analysis of whole-cell and exported proteins; Thomson-Carter FM et al.; Thirty-four isolates of methicillin-resistant Staphylococcus aureus (MRSA) from patients in Glasgow Royal Infirmary were studied . Whole-cell protein profiles obtained by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) were compared with banding patterns produced by immunoblots of exported proteins . Human plasma was used as a source of staphylococcal antibodies for the immunoblots . SDS-PAGE of whole-cell extracts did not usefully distinguish different isolates of MRSA . Reproducible banding patterns were obtained by immunoblots of exported proteins . Analyses of immunoblots by use of the Dice coefficient demonstrated that isolates of MRSA could be divided into two main groups . Immunoblots of exported proteins provided a rapid, reproducible and sensitive method for characterisation of MRSA.

J Med Microbiol, 1989 Jan, 28(1), 15 - 23
A new methicillin- and gentamicin-resistant Staphylococcus aureus in Dublin: molecular genetic analysis; Carroll JD et al.; In June 1985 two new strains of methicillin- and gentamicin-resistant Staphylococcus aureus (MGRSA) were isolated in a Dublin hospital . Of these, one strain spread rapidly, affecting a total of 65 patients during the following 18 months, and subsequently spread to a second Dublin hospital . Detailed laboratory studies, including plasmid screening, plasmid restriction enzyme digest pattern analysis, hybridisation analysis, location of resistance determinants, and bacteriophage typing with a set of experimental S . aureus typing phages, demonstrated that the "new" MGRSA organisms, termed Phenotype III Dublin isolates, were completely distinct from, and unrelated to, the MGRSA strains responsible for serious nosocomial infections in Dublin hospitals during the decade before June 1985 . These Phenotype III isolates were very similar to MGRSA organisms isolated in a Baghdad hospital during 1984 . Data from plasmid curing, plasmid transfer and hybridisation experiments indicated that 20% of the Phenotype-III isolates expressed chromosomally encoded, high level resistance to ethidium bromide (MIC 120 micrograms/ml), and that this was possibly due to chromosomal integration of a penicillinase-like plasmid.

Proc Soc Exp Biol Med, 1989 Jan, 190(1), 79 - 86
Evaluation of an anti-inflammatory factor derived from hyperimmunized cows; Owens WE et al.; An anti-inflammatory factor isolated from milk of hyperimmunized cows was analyzed in vitro and in vivo . Macrophages collected from lacteal secretions of a unimmunized nonlactating cow showed increased ability to kill phagocytosed Staphylococcus aureus when incubated with the anti-inflammatory factor . Mice injected intraperitoneally with 10 mg/kg of anti-inflammatory factor demonstrated an increased LD50 to S . aureus when challenged intraperitoneally . Injected mice also demonstrated significantly (P less than 0.05) less mammary inflammation and involution and increased clearance of S . aureus when challenged intramammarily . Quantitative histologic analysis of mammary tissues from mice injected with anti-inflammatory factor demonstrated significantly (P less than 0.05) more lumen, less interalveolar connective tissue, and less leukocytic infiltration compared with control mice . Mammary glands of mice injected with anti-inflammatory factor and challenged with S . aureus also contained fewer colony-forming units than control mice . The product appeared to exert its effect on the nonspecific defense system via modulation of leukocyte function.

Clin Immunol Immunopathol, 1989 Jan, 50(1 Pt 1), 100 - 8
Effects of retinoids on human thymus-dependent and thymus-independent mitogenesis; Dillehay DL et al.; The effects of all-trans-retinoic acid (RA), 13-cis-retinoic acid, and N-(4-hydroxyphenyl)retinamide on the mitogenic responses of various populations of human lymphocytes have been evaluated . Superoptimal concentrations of mitogens allowed the greatest RA-induced potentiation of lymphocyte proliferation . All three retinoids at concentrations as low as 5 x 10(-14)M significantly potentiated the proliferation of adenoidal and tonsillar lymphocytes stimulated by pokeweed mitogen (PWM) . However, the responses of adenoidal and tonsillar lymphocytes to Staphylococcus aureus Cowan strain A were not potentiated by retinoids . Retinoids also caused significant potentiation of proliferation of PWM-stimulated peripheral blood lymphocytes (PBL) . However, endpoint concentrations of retinoids required to significantly potentiate PBL proliferative responses to PWM were much higher than required for potentiation of adenoidal or tonsillar lymphocytes . PBL responses to concanavalin A (Con A) were significantly potentiated by retinoid concentrations as low as 10(-8) to 10(-10) M . Retinoid-potentiated responses were also observed wi Con A-stimulated thymocytes, but the endpoint concentrations required for significant potentiation were 10-fold higher than required to potentiate PBL responses to Con A . These data indicate that the sensitivity of lymphocytes to the retinoid-mediated potentiation of mitogenesis depends on the lymphoid compartment from which the cells are obtained . Tonsillar and adenoidal lymphocytes were the most responsive of the lymphocytes tested to the retinoid-induced potentiation of PWM responses . In addition, retinoids appear to selectively potentiate T cell-dependent proliferative activity.

Am J Dis Child, 1989 Jan, 143(1), 34 - 9
Epidemic methicillin-gentamicin-resistant Staphylococcus aureus in a neonatal intensive care unit; Reboli AC et al.; Between October 1985 and August 1986, 49 isolates of methicillin-resistant Staphylococcus aureus (MRSA) were obtained from 26 neonates in the neonatal intensive care unit (NICU) at the Medical University Hospital, Charleston, SC . Sites of MRSA isolation were the respiratory tract (33%); nasopharynx (12%); gastrointestinal tract (12%); eye (8%); blood (6%); and catheter tips, wounds, or umbilicus (29%) . Very low birth weight was a significant risk factor for MRSA acquisition . All isolates had the same phage type (47/54/75/83A), antibiogram, and whole-cell protein profile . Agarose gel electrophoresis of all 49 isolates disclosed a plasmid level of approximately 45 X 106 daltons (45 megadaltons) in ten different isolates and no plasmid DNA in 39 isolates . Cultures of NICU personnel failed to disclose MRSA carriers and environmental cultures for MRSA were negative . Ten selected isolates showed lower minimal bactericidal concentrations for hexachlorophene than for chlorhexidine . Standard infection-control measures such as contact isolation, hand washing with chlorhexidine, and cohorting (when possible) failed to contain the epidemic . Ultimately, eradication of MRSA from the NICU was associated with the institution of hexachlorophene hand washing.

Infect Immun, 1989 Jan, 57(1), 231 - 4
Determination by western blot (immunoblot) of seroconversions to toxic shock syndrome (TSS) toxin 1 and enterotoxin A, B, or C during infection with TSS- and non-TSS-associated Staphylococcus aureus; Whiting JL et al.; Serum antibody responses to toxic shock syndrome (TSS) toxin 1 (TSST-1) and staphylococcal enterotoxins A, B, and C were determined by western blot (immunoblot) analysis of acute- and convalescent-phase paired sera from 18 TSS- and 31 non-TSS-associated Staphylococcus aureus infections . Compared with non-TSS cases, seroconversion to TSST-1 was significantly more frequent among both menstrual (5 of 8 versus 1 of 31; P less than 0.001) and nonmenstrual (3 of 10; P less than 0.05) patients . Seroconversion to staphylococcal enterotoxin A was also more frequent among both menstrual (2 of 8 versus 0 of 31; P less than 0.05) and nonmenstrual (2 of 9; P less than 0.05) TSS patients . In general, patients with TSS associated with TSST-1-positive S . aureus were more likely to seroconvert exclusively to TSST-1 (4 of 12 versus 0 of 6; P = 0.16), whereas those associated with TSST-1-negative S . aureus were more likely to seroconvert exclusively to enterotoxins (3 of 6 versus 0 of 11; P less than 0.05) . Concurrent seroconversions to multiple exoproteins were more frequent among both menstrual (3 of 8; P less than 0.05) and nonmenstrual (2 of 9; P less than 0.05) TSS patients compared with persons without TSS (0 of 31) . These data suggest but do not prove that enterotoxins (especially staphylococcal enterotoxin A) in addition to TSST-1 may be involved in both menstrual and nonmenstrual TSS . Furthermore, since exposure to multiple exoproteins is more likely to occur during TSS-associated than non-TSS-associated S . aureus infections, the possibility of additive or synergistic effects of these putative toxins in the pathogenesis of TSS should be further explored.

J Hyg Epidemiol Microbiol Immunol, 1989, 33(3), 277 - 82
Incidence of enterotoxin producing Staphylococcus aureus among pyogenic skin infections; Naidu S et al.; Out of the total 68 S . aureus strains isolated and studied from pyoderma patients, 33 (48.5%) strains produced enterotoxin . Isolates from IED, impetigo and folliculitis exhibited high degree of enterotoxigenicity . SE-C and its combinations with other enterotoxins was common . 60.6% of the SE producers were found phage nontypable . Typable enterotoxigenic strains were associated with III, IV and mixed phage groups . S . aureus var . hominis and S . aureus var . bovis are the prevalent subspecies types and potent SE producers among pyogenic skin infections.

Acta Derm Venereol Suppl (Stockh), 1989, 144, 83 - 7
IgE antibody to sweat in atopic dermatitis; Adachi K et al.; Of 45 patients with atopic dermatitis skin-tested with their own sweat, 43 showed positive immediate-type skin reactions to titres between 1 and 256 . Of 22 non-atopic patients 18 showed negative reactions . Skin reactivity of the atopic patients to the sweat and house dust did not run parallel . Radioallergosorbent test (RAST) using the sweat collected from a healthy subject detected IgE antibody in 24 atopic patients with a score from 0.5 to 3.5, whereas all the control subjects showed the score 0 . This IgE antibody to sweat did not cross-react with the mite extract (Dermatophagoides farinae) or Staphylococcus aureus . These results indicate that atopic patients have specific IgE antibody to sweat.

Chemotherapy, 1989, 35(5), 345 - 50
Paradoxical effect of cloxacillin and benzylpenicillin against clinical isolates in Staphylococcus aureus; Odenholt I et al.; The occurrence of a paradoxical effect of cloxacillin and benzylpenicillin in 7 strains of Staphylococcus aureus was studied with time killing curves in vitro . The isolates were exposed to 1, 2, 10 and 100 x MIC of the antibiotics . A paradoxical effect was found in 5/7 strains with cloxacillin and in 4/7 with benzylpenicillin . All strains were killed faster with 2 x MIC than with 100 x MIC . All strains that showed a paradoxical effect with cloxacillin did so already at 10 x MIC . For benzylpenicillin the effect occurred at 10 x MIC for 2 strains and at 100 x MIC for 2 . A single MBC/MIC ratio will not reveal the paradoxical effect.

Scand J Infect Dis, 1989, 21(3), 343 - 4
Tropical pyomyositis in a Greek adult; Meletis J et al.; A case of tropical pyomyositis in a 24-year-old Greek is reported . The patient presented with high fever and swelling of the left thigh, generalized lymphadenopathy and multiple infiltrations in both lung fields on X-ray of the chest . Multifocal muscle abscesses were detected by CT scan of the left thigh and gluteal area . Staphylococcus aureus was identified in cultures of the purulent material which was surgically drained . The patient was subsequently treated with appropriate antibiotics . Lack of familiarity with this disease caused diagnostic confusion and delayed the initiation of treatment.

Arch Dermatol Res, 1989, 281(1), 31 - 4
Analysis of the proinflammatory property of epidermal cyst contents: chemotactic C5a anaphylatoxin generation; Terui T et al.; We investigated in vitro the contents of epidermal cysts for complement activation and found that they activated complement mainly through the alternative pathway . Chemotactic C5a anaphylatoxin produced by the cyst contents after contact with serum most likely plays a significant role in the initiation and aggravation of inflammation in ruptured epidermal cysts . Our additional study disclosed that components of three representative follicular resident microorganisms (Pityrosporum ovale, Propionibacterium acnes, and Staphylococcus epidermidis) also produced C5a anaphylatoxin mainly through the alternative pathway; the C5a production was more vigorous than that by a virulent pathogen, Staphylococcus aureus . These results suggest that accidental colonization of the cyst contents by these follicular microbial flora further augments the inflammatory changes in ruptured epidermal cysts.

Wiad Parazytol, 1989, 35(2), 121 - 6
{Observation of the processes of phagocytosis and digestion of Staphylococcus aureus by Trichomonas vaginalis using a transmission electron microscope}; Szreter H et al.; Trichomonas vaginalis was incubated with bacteria for 30 min . After centrifugation of Staphylococcus aureus, incubation of T . vaginalis was continued for 40 min., 120 min . and 6 hrs . It was found that bacteria have been intensively phagocytated through whole surface of protozoan . Phagosome with bacteria at different stages of degradation, up to defecation of undigested residues were observed . It was observed that some bacteria, apparently not susceptible to digestion, were immersed directly in cytoplasm (outside vacuoles).

Diagn Microbiol Infect Dis, 1989 Jan-Feb, 12(1), 57 - 65
Evaluation of cefotaxime alone and in combination with desacetylcefotaxime against strains of Staphylococcus aureus that produce variants of staphylococcal beta-lactamase; Stratton CW et al.; We evaluated cefotaxime (CTX) alone and in combination with its metabolite, desacetylcefotaxime (dCTX) against strains of Staphylococcus aureus that produce the four recognized variants of staphylococcal beta-lactamase and a beta-lactamase-producing isolate characterized by the expression of borderline resistance to methicillin . Although macrodilution MICs revealed that dCTX was less active than CTX against these strains (geometric means of 16 micrograms/ml and 4 micrograms/ml, respectively), the addition of clinically achievable concentrations of dCTX to CTX resulted in a reduction in the observed CTX MICs . This effect was similar to although less pronounced than that obtained by combining clavulanic acid with cefazolin . The increased antistaphylococcal activity noted by MIC determinations was confirmed with kill-kinetic studies . Determination of the relative rates of hydrolysis of selected cephalosporins showed that neither CTX nor dCTX were appreciably hydrolyzed by the variant staphylococcal enzymes . Evaluation of the effect of CTX and dCTX upon the staphylococcal beta-lactamases demonstrated that neither agent inhibited the destruction of a 100 microM solution of nitrocefin, although the reduction of CTX and cefazolin MICs by low concentrations of dCTX suggests that the dCTX metabolite may act as a competitive inhibitor of beta-lactamase . These observations may explain the previously demonstrated clinical efficacy of CTX used alone for the treatment of serious infections caused by S . aureus.

Res Vet Sci, 1989 Jan, 46(1), 84 - 9
Ovine opsonins for Staphylococcus aureus cell wall and pseudocapsule; Watson DL; Blood serum, milk whey and purified IgG1 and IgG2 antibody preparations from sheep immunised with live or killed Staphylococcus aureus vaccines were assayed for opsonising activity against S aureus using ovine mammary neutrophils as effector cells . Serum had much higher levels of opsonins than whey; whey did not inhibit the potent opsonising activity of hyperimmune serum . IgG1 antibodies against cell wall antigens of in vitro-grown S aureus and pseudocapsular antigens of in vivo-grown S aureus had low opsonising activity . IgG2 antibody against in vitro-grown S aureus surface antigens had significantly greater opsonising activity than IgG1 of this specificity . The most powerful opsonising activity was associated with IgG2 antibody against the pseudo-capsular antigens produced by S aureus grown under in vivo, or simulated in vivo cultural conditions.

Arch Biochem Biophys, 1989 Jan, 268(1), 338 - 48
Isolation of a Staphylococcus aureus beta-lactamase-dicloxacillin complex and kinetic studies on the reactivation of the enzyme; Hardy LW et al.; Exposure of the beta-lactamase from Staphylococcus aureus to the slowly reacting substrates cloxacillin or dicloxacillin results in time-dependent inactivation of the enzyme . Methods for the rapid separation of a beta-lactamase-dicloxacillin complex from excess inhibitor, using centrifuged columns of Sephadex G-25 or DEAE-Sephadex G-25, are described . The enzyme-dicloxacillin complex releases active enzyme, with specific activity identical to that of untreated enzyme, after storage at pH 7.5 at 15 degrees C . Full reactivation was accompanied by the release of 0.8 eq of hydrolyzed dicloxacillin . The complex is stable for up to 40 h when stored at pH 3 at 4 degrees C . The reactivation process, which occurs with first-order kinetics at 15 degrees C and pH values between 4 and 8, displays a pH dependence with apparent pKa's of 4.6 and 8.5, and a limiting value of the reactivation rate constant of 0.022 min-1 . Deviation from first-order kinetics at pH 9 is consistent with a competing irreversible inactivation of the enzyme at that pH . This behavior differs substantially from that of the similarly inactivated beta-lactamase I from Bacillus cereus, whose rate of reactivation is independent of pH, but which undergoes irreversible denaturation at acidic pH {A . L . Fink, K . M . Behner, and A . K . Tan (1987) Biochemistry 26, 4248-4258} . Addition of hydroxylamine to the S . aureus beta-lactamase-dicloxacillin, complex stimulates the rate of reactivation by a maximum of 35% . This effect is hyperbolically dependent on the concentration of hydroxylamine with half-maximal stimulation at 2.8 mM . The Km for ampicillin hydrolysis catalyzed by the partially reactivated enzyme is identical to that measured for catalysis by the untreated enzyme . We discuss our observations in relation to models for the transient inhibition process.

Infect Immun, 1989 Jan, 57(1), 48 - 54
Enhancement of antibacterial resistance of neutropenic, bone marrow-suppressed mice by interleukin-1 alpha; McIntyre KW et al.; The effect of recombinant human interleukin-1 alpha (IL-1) on the resistance of normal and bone marrow-suppressed mice against bacterial infection was evaluated . IL-1 induced neutrophilia and enhanced the resistance of normal mice against acute, systemic intraperitoneal infection with Klebsiella pneumoniae and methicillin-resistant Staphylococcus aureus . Mice with cyclophosphamide-induced bone marrow suppression were neutropenic and exhibited increased susceptibility to infection . Treatment of neutropenic C57BL/6 and C3H/HeJ mice with IL-1 before infection accelerated recovery of peripheral neutrophil counts and stimulated resistance against infection . Increases in neutrophils and enhancement of resistance induced by IL-1 were both dose and time dependent . Both neutrophilia and augmented resistance to infection were eliminated by a second dose of cyclophosphamide administered during the IL-1 treatments . Bone marrow-suppressed mice treated with IL-1 showed, at 4 h postinfection, greater increases in peripheral blood neutrophils and in numbers of peritoneal exudate neutrophils than suppressed mice treated with vehicle . The data suggest that the IL-1-stimulated recovery of myelopoiesis is an important factor in the enhancement of antibacterial resistance in bone marrow-suppressed, neutropenic mice . These findings indicate that IL-1 may be efficacious in limiting the duration of the neutropenia and of the increased risk for the development of bacterial infection associated with bone marrow suppression.

AJR Am J Roentgenol, 1989 Jan, 152(1), 131 - 6
Dyke award . Gd-DTPA-enhanced MR imaging of experimental bacterial meningitis: evaluation and comparison with CT; Mathews VP et al.; Gd-DTPA-enhanced MR images of experimental bacterial meningitis were obtained after Staphylococcus aureus was inoculated directly into the cisterna magna of four dogs . Each animal was studied with both unenhanced and enhanced MR and CT with Gd-DTPA and meglumine iothalamate, respectively . The enhancement patterns resulting from these techniques were compared and images were correlated with histopathology . All animals demonstrated abnormal leptomeningeal enhancement on MR with Gd-DTPA, but only one of four dogs exhibited abnormal contrast enhancement on CT . In these animals Gd-DTPA-enhanced MR also identified complications of meningitis, such as ventriculitis and cerebritis, more effectively than CT did . Unenhanced MR was not helpful in identifying meningitis . Histologic evaluation demonstrated that the abnormal areas of contrast enhancement on MR and CT correlated with inflammatory cell infiltration . However, some regions of mild leptomeningitis, ependymitis, and cerebritis identified histologically did not demonstrate abnormal enhancement . Since the animal model used was clinically and pathologically similar to human meningitis, we propose that Gd-DTPA-enhanced MR will subsequently be found more effective than unenhanced MR and IV contrast-enhanced CT for demonstrating meningitis and its complications in humans.

J Gen Microbiol, 1989 Jan, 135 ( Pt 1), 1 - 10
Physical and biochemical characterization of the qacA gene encoding antiseptic and disinfectant resistance in Staphylococcus aureus; Tennent JM et al.; We have previously cloned a 3.5 kb fragment from the Staphylococcus aureus multiresistance plasmid pSK1 which carries the qacA determinant responsible for linked resistance to acriflavine (Acr), ethidium bromide (Ebr), quaternary ammonium compounds (Qar), propamidine isethionate (Pir), and diamidinodiphenylamine dihydrochloride (Ddr) . This report presents a biochemical and physical analysis of qacA and shows the widespread carriage of this gene on S . aureus resistance plasmids . Tn5 insertion mutagenesis defined the extent of qacA to within 2.40 kb of pSK1 DNA . Examination of the expression of insertion and deletion mutants of the cloned qacA sequences in both maxicells and minicells led to the association of a 50 kDa protein, designated QacA, with the AcrEbrQarPirDdr phenotype . Based on fluorimetric and isotopic assays used to determine the extent of accumulation of ethidium bromide by S . aureus strains harbouring pSK1, we propose that the basis of AcrEbrQarPirDdr in S . aureus is a qacA-mediated efflux system.

J Antimicrob Chemother, 1989 Jan, 23(1), 79 - 86
Aminoglycosides enhance the adherence of Staphylococcus aureus to HeLa cells; Miyake Y et al.; Sub-lethal concentrations of aminoglycosides enhanced the adherence of Staphylococcus aureus FDA 209P to HeLa cells, whereas beta-lactams, pyridone carboxylic acid derivatives and chloramphenicol did not . Treatment with aminoglycosides also enhanced the bacterial adherence of these cells to immobilized fibronectin and laminin, and changed the bacterial cell surface to a more hydrophilic state than exists in non-treated cells . The adherence of S . aureus was not inhibited by lipoteichoic acid, extracted from the same strain, regardless of whether the bacterial cells were treated with an aminoglycoside or not . No correlation was observed between the adherence and zeta-potential of S . aureus.

Nephron, 1989, 52(4), 307 - 12
Effect of ciclosporin on lymphocyte subpopulations and immunoglobulin production in IgA nephropathy; Lai KN et al.; The effect of ciclosporin on the cellular immunoregulation was examined in 19 patients with IgA nephropathy . The patients were randomly divided into two groups: 9 patients receiving oral ciclosporin (5 mg/kg/day) for 12 weeks and 10 patients receiving placebo and acting as controls . T lymphocyte subpopulations were determined using OKT monoclonal antibodies . The functional capability of lymphocytes was assessed by in vitro immunoglobulin synthesis of cultured peripheral mononuclear cells, thymidine uptake by cultured lymphocytes, and t lymphocyte activation with expression of interleukin-2 receptors . A fall of in vitro IgA production by cultured lymphocytes (p less than 0.05), a reduction of thymidine uptake by Staphylococcus aureus Cowan-stimulated cultured lymphocytes (p less than 0.05), and a reduction of activated lymphocytes expressing interleukin-2 receptor (p less than 0.05) were observed in patients after 12 weeks of ciclosporin therapy . The percentages of OKT4 and OKT8 lymphocytes and OKT4/8 ratios were not altered with therapy . A simultaneous reduction of proteinuria and a transient impairment of renal function were observed . Similar changes in cellular immune parameters and clinical response were not observed in the controls . Our study suggests ciclosporin could modulate the cellular immunity in IgA nephropathy.

Microbios, 1989, 58(236-237), 147 - 54
Influence of cefotaxime on Staphylococcus aureus with respect to their interaction with macrophages; Cuffini AM et al.; Many antibiotics in addition to their direct effect on bacteria can modulate host defence in different ways . The influence of cefotaxime on the phagocytic and bactericidal activity of mouse peritoneal macrophages was investigated . At concentrations of half the MIC the antibiotic caused macrophages to kill intracellular Staphylococcus aureus at a higher level than did macrophages without drug . Bacteria pretreated with cefotaxime became more sensitive to macrophage phagocytic and bactericidal activity . Macrophages taken from mice receiving intravenous cefotaxime showed greater phagocytic capacity than those from control mice.

Med Dosw Mikrobiol, 1989, 41(1), 1 - 8
{Effect of cadmium on C-14-glucose uptake by Staphylococcus aureus}; Tynecka Z et al.; The aim of the study was to evaluate an influence of Cd++ on 14C-glucose uptake by two strains of S . aureus resistant and sensitive to cadmium in 0.1 M phosphate buffer, pH 7.0 . Uptake of this sugar in both strains is an active process in which energy comes from oxidation of endogenous substrates, what was shown in aerobic condition, anaerobic condition at temperatures of 37 degrees C and 4 degrees C, and with p-CMB and CCCP . In the resistant strains Cd++ at 10 microM concentration did not inhibit endogenous respiration, 14C-glucose uptake and its oxidation . This is due to presence of energy-dependent system of 2H+/Cd++ antiport coded by cadA genes located on penicillinase pII17810 plasmid, which eliminated Cd++ from bacterial cell . In the case of plasmid free variant deprived of this system, Cd++ is retained in cytoplasm and blocks endogenous respiration uptake, and oxidation of glucose.

Microbios, 1989, 58(234), 27 - 33
Comparative effects of roxithromycin and erythromycin on cellular immune functions in vitro . 3 . Killing of intracellular Staphylococcus aureus by human macrophages; Cuffini AM et al.; The in vitro effect of roxithromycin on the bactericidal activity of human macrophages was compared with that of erythromycin . At a concentration of half the MIC roxithromycin showed a much more impressive killing capacity on macrophage-ingested live Staphylococcus aureus than did erythromycin, under all the experimental conditions used.

J Orthop Res, 1989, 7(4), 543 - 9
Three-phase {99mTc}diphosphonate scintimetry in septic and nonseptic arthritis of the immature knee: an experimental investigation in dogs; Hansen ES et al.; We compared the informative value of dynamic and static {99mTc}diphosphonate ({99mTc}DPD) scintimetry in early septic arthritis (SA) and chronic nonseptic arthritis (NSA) of the knee in puppies . SA (n = 10), induced by injection of Staphylococcus aureus into one knee, was examined after 48 h . NSA (n = 6) was induced by weekly intraarticular instillation of 1% carrageenan and examined at 2 and 12 weeks . Epiphyseal and metaphyseal count ratios (CRs) between experimental and control joint were calculated in an angiographic phase (0-20 s), a "blood pool" phase (20-256 s), and a bone uptake phase (2 hs) . Control dogs (n = 4) had a CR of 1.0 in all regions and phases, the coefficients of variation being 0.06 and 0.03 in dynamic and static phases, respectively . In NSA, all scintimetric phases showed decreased metaphyseal uptake and largely unchanged epiphyseal uptake . Generally, SA exhibited vastly increased angiographic uptake in all regions and increased epiphyseal blood pool uptake, whereas delayed epiphyseal uptake varied and delayed metaphyseal uptake tended to decrease . Two septic joints had generalized reduction in dynamic uptake, probably owing to increased intraarticular pressure; in one of these, dynamic uptake in the distal femoral epiphysis (FE) was totally absent, suggesting temporary circulatory arrest during the dynamic scanning procedure . The use of regional dynamic and static {99mTc}DPD scintimetry increased the pathophysiologic and diagnostic value of joint scintigraphy, especially with respect to the early detection of SA and its avascular complications.

Drugs Exp Clin Res, 1989, 15(2), 63 - 9
Phagocytosis and killing of A-protein positive Staphylococcus aureus in the presence of low doses of antibiotics; Mascellino MT et al.; An A-protein positive strain of Staphylococcus aureus was grown in varying concentrations of erythromycin, clindamycin and miocamycin at sub-MIC levels and incubated in purified human polymorphonuclear leukocytes (PMNs) . The presence of A-protein produced resistance of the above strain to phagocytosis and killing by PMNs and a slight resistance to opsonization by normal serum . The addition of antibiotics, which inhibit protein biosynthesis, caused significant changes in the ability of this bacterium to resist opsonization by serum complement . The antimicrobial agents produced a significant enhancement of bacterial killing and phagocytosis . In particular, clindamycin and erythromycin, at sub-MIC concentrations, modified the hair-like structure of S . aureus, which contains A-protein, and were able to affect the interaction of the strain with phagocytic cells.

Scand J Infect Dis, 1989, 21(1), 95 - 101
Comparison of in vivo and in vitro activities of antibiotics with different modes of action against a tolerant and a non-tolerant Staphylococcus aureus strain; Hoogeterp JJ et al.; The antibacterial efficacies of 4 antibiotics with different modes of action against a penicillin-tolerant and a non-tolerant strain of Staphylococcus aureus were investigated . For the in vitro studies the minimum inhibitory concentration (MIC) and the minimum bacterial concentration (MBC) were determined and short-term growth experiments at different antibiotic concentrations were performed . For the in vivo studies, antibacterial efficacy in an experimental infection in normal and granulocytopenic mice was evaluated . For erythromycin, rifampicin and ciprofloxacin, there was no difference in the MIC and MBC values for the 2 strains . Benzylpenicillin had an MBC value for the tolerant strains which was 256 times higher than the MIC; with the non-tolerant strain there was no difference . EC50 values, calculated from the in vitro short-term growth curves, gave similar results . Only benzylpenicillin exhibited a difference in activity against the tolerant strain, as reflected by the EC50 that was 290 times the EC50 for the non-tolerant strain . Studies in normal and granulocytopenic mice gave similar results: benzylpenicillin was 268 times less active against the tolerant strain than against the non-tolerant strain . Erythromycin, rifampicin and ciprofloxacin were 2-3 times less active against the tolerant strain than against the non-tolerant strain . The presence of granulocytes is important for the antibacterial effect of all antibiotics studied, since in the absence of granulocytes higher doses of the antibiotics are needed in order to obtain the same antibacterial effect as when granulocytes are present.

Microbiol Immunol, 1989, 33(3), 175 - 82
Improved methods for detection and serotyping of coagulase from Staphylococcus aureus; Hwang SM et al.; Improved methods for detection and serotyping of staphylocoagulase were concomitantly devised . We devised an improved method for detection of coagulase activity on agarose film in the same manner as single radial immunodiffusion . The amounts of reagents required for detection of coagulase on agarose film were successfully diminished by adding polyethylene glycol (PEG) to the original formula described by Boothby et al . Using microplates in another improved method for coagulase serotyping, the amount of reaction fluid required was considerably less compared with the conventional tube method . PEG was found to be also effective to increase the efficacy of coagulase serotyping . In the presence or absence of anti-coagulase antisera, culture supernatants of staphylococcal strain grown in brain heart infusion broth were incubated with the reaction fluid containing bovine fibrinogen, rabbit plasma, 6-amino caproic acid, polyethylene glycol 6,000 . Coagulase activity was visualized as a turbid mass formed in the wells . Turbid mass formation due to coagulase activity was type-specifically inhibited in the presence of type-specific antisera . Detailed procedures of the methods are precisely described with some preliminary results obtained by the methods.

Vopr Pitan, 1989 Jan-Feb, (1), 60 - 5
{Simulation of the Baird-Parker media formula based on Soviet ingredients for the isolation of Staphylococcus aureus}; Petrushina LI et al.; A medium has been prepared based on Soviet nutrient agar and reagents supplemented by ingredients from Baird-Parker's medium . Its growth and selective properties are close to those of Baird-Parker's medium . The new selective medium has been recommended for isolation of S . aureus from food products and environment objects.

Arch Fr Pediatr, 1989 Jan, 46(1), 41 - 3
{Portal system obstruction of delayed onset following neonatal Staphylococcus aureus infection}; Garel D et al.; Two neonates underwent staphylococcal septicemia with multiple intrahepatic abscesses, following umbilical catheterization in one case and due to superinfected heel puncture in the other case . At the first examination, liver ultrasonography showed multiple hypoechogenic areas and assessed patency of the portal vein flow . In spite of clinical, biological and sonographic recovery within the following weeks, portal hypertension due to seemingly late portal vein thrombosis occurred . These clinical reports indicate the need for a protracted echosonographic supervision of neonatal intrahepatic abscesses and portal vein patency before asserting complete recovery.

Antimicrob Agents Chemother, 1989 Jan, 33(1), 128 - 30
Ciprofloxacin for methicillin-resistant Staphylococcus aureus infections; Piercy EA et al.; Thirty-seven patients with methicillin-resistant Staphylococcus aureus infections and/or colonization were treated with oral ciprofloxacin (750 mg twice a day) . Clinical cure or improvement of infections occurred in 91% of the patients, and bacteriologic cure occurred in 60% . Ciprofloxacin therapy suppressed methicillin-resistant S . aureus colonization in 55% of the patients . Ciprofloxacin-resistant strains emerged in 6 of the 37 patients.

Antimicrob Agents Chemother, 1989 Jan, 33(1), 120 - 1
Interaction of FCE 22101 with penicillin-binding proteins of Staphylococcus aureus; Tonin EA et al.; FCE 22101 is a new penem characterized by a broad spectrum of activity which includes activity against methicillin-resistant strains of Staphylococcus aureus . The interaction of FCE 22101 with penicillin-binding protein 2a of a methicillin-resistant S . aureus strain has been investigated in the present study . In competition experiments, the penem showed a very low affinity for this protein, and a concentration more than 4 times the MIC was required for penicillin-binding protein 2a saturation . When the classical competitive procedure was modified by increasing the time of incubation of either membranes or growing cells with FCE 22101, the antibiotic showed a much higher affinity for penicillin-binding protein 2a and saturated the protein at a concentration close to the MIC, with slow kinetics.

Acta Neurochir (Wien), 1989, 96(3-4), 141 - 8
Effect of dexamethasone on various stages of experimental brain abscess; Yildizhan A et al.; Rats were inoculated with staphylococcus aureus to produce cerebral abscesses and treated with either antibiotic or dexamethasone and with antibiotic plus dexamethasone at sequential stages of abscess formation . Antibiotic alone shortened the cerebritis stage, accelerated the encapsulation and affected the bacterial clearance in the abscess centre when it was started early in the course of cerebritis . Dexamethasone impaired the lymphocytic and fibroblastic responses and delayed the collagen deposition as well as suppressed the efficacy of antibiotic . However, it did not halt entirely the encapsulation and did reduce the associated cerebral oedema.

Vet Immunol Immunopathol, 1989 Jan, 20(2), 149 - 63
In vivo effects of a thymosin alpha 1-containing colostral whey product on neutrophils and lymphocytes from lactating cows without and with experimentally induced Staphylococcus aureus mastitis; Kehrli ME Jr et al.; Two separate experiments evaluated ID-1 (a commercial bovine whey product containing 5200 pg of thymosin alpha 1/ml) as an immunotherapeutic agent in lactating cows . In the first experiment, cows without mastitis were evaluated for blood leukogram, milk production, total and differential milk cell counts, lymphocyte (Lc) blastogenesis, and neutrophil (PMN) functions (random and directed migration under agarose, chemiluminescence, ingestion of bacteria, iodination, cytochrome C reduction, antibody-independent neutrophil-mediated cytotoxicity, and antibody-dependent cell-mediated cytotoxicity) before and after ID-1 therapy . ID-1 treatment resulted in a significant treatment group by time period interaction for the relative proportion of mononuclear cells (MNC) in milk (P less than 0.009) and for PMN random migration (P less than 0.01) . Based on these interactions, ID-1 treatment appeared to slightly increase the proportion of small MNC in milk and to increase random migration from pretreatment levels by 73% more than increases observed in controls . No significant effect of ID-1 treatment on milk production, total milk somatic cell counts, Lc blastogenesis, or other PMN functions was observed . In cows with experimental Staphylococcus aureus intramammary infections, ID-1 treatment resulted in a significant decline in blood leukocyte count (P less than 0.001) and blood PMN count (P less than 0.02), and maintained PMN random migration (P less than 0.01) while controls declined and abrogated a depression in the ability of Lc to respond to mitogens (P less than 0.05) that developed in controls as a result of S . aureus mastitis . Injection of ID-1 into cows had no adverse effect on their overall health or level of milk production, but did cause subtle and potentially favorable changes in several in vitro immune parameters . In spite of these subtle changes which might indicate increased resistance to mastitis, cows actually developed a more severe S . aureus intramammary infection based on a 9% increase in log 10 bacterial shedding in milk.

Indian J Pathol Microbiol, 1989 Jan, 32(1), 57 - 61
Anaerobic bacteria in brain abscess, a review; observation of 18 consecutive cases; Narayan K et al.; The aerobic and anaerobic bacterial flora of 18 consecutive cases of brain abscesses were studied . Two cases were sterile . In five cases there was a single bacterial isolate . In five cases there was a single bacterial isolate the organism being Staphylococcus aureus in two cases, Bacteroides fragilis in two cases and Peptostreptococcus anaerobius in one case . In the remaining eleven cases, more than one organism was isolated . A combination of anaerobe with an aerobe was observed in five cases and infections with more than one anaerobe in as many as six cases . Anaerobic organisms are the important pathogens in brain abscess and occurrence of infections by more than one organism is a common factor.

Mol Biother, 1989, 1(4), 186 - 207
Staphylococcal protein A adsorption in neoplastic disease: analysis of physicochemical aspects; Murphy RM et al.; Staphylococcus aureus organisms and immobilized Staphylococcal protein A have been used as extracorporeal immunoadsorbents for the treatment of neoplastic disease . In a significant number of cases, a biologic response has been documented . This review analyzes the experimental results to date, correlates experimental design parameters with response to treatment, and discusses possible mechanisms of action . The evidence implicates protein A as the tumoricidal agent, although bacterial or complement components may act independently or synergistically . A more quantitative and fundamental approach is needed to exploit the potential for protein A immunoadsorption in treating neoplastic disease.

Drugs Exp Clin Res, 1989, 15(4), 141 - 4
The influence of cefepime on opsonophagocytosis of bacteria; Raponi G et al.; The effect of subminimal inhibitory concentrations (sub-MICs) of a semi-synthetic beta-lactam antibiotic, cefepime (BMY 28142), was studied on the opsonophagocytosis by human polymorphonuclear leukocytes (PNMs) of strains of Escherichia coli, Klebsiella oxytoca and Staphylococcus aureus . Cefepime-treated Gram-negative bacteria were not better phagocytosed by PMNs; however the uptake of one of three S . aureus strains was enhanced . The effect of cefepime on the uptake of strains of E . coli was compared to that of two other beta-lactam antibiotics, mecillinam and ceftriaxone . Exposure of E . coli to sub-MICs of mecillinam and ceftriaxone enhanced phagocytosis by PMNs.

Rev Cubana Med Trop, 1989 Jan-Apr, 41(1), 114 - 8
{Isolation of beta lactamase enzyme in strains of Staphylococcus aureus with the method of starch paper}; Prat Lariot A et al.; The authors worked with 2,273 strains of Staphylococcus aureus isolated from clinical samples processed at the Microbiology laboratory of Centro Habana Pediatric Teaching Hospital in the period ranging from June 1985 to December 1986 . All strains underwent the bacterial sensitivity test against antibiotics by Bauer-Kirby technique . Fourty-eight strains were sensitive to penicillin - which accounts for 2.11% - by the beta-lactamase detection test by the starch paper method . In all cases, negative results were obtained, which is in agreement with the results of the sensitivity tests.

J Singapore Paediatr Soc, 1989, 31(1-2), 75 - 8
Bacterial liver abscess in children; Wang DS et al.; From November 1987, 136 children with bacterial liver abscess were encountered . There were 97 males and 39 females and the age ranged from 1-15 years (mean = 8.42 years) . The clinical signs and symptoms of liver abscess were confirmed by radiography, ultrasonography, percutaneous transhepatic drainage (PTHD) and radioisotope scanning . 103 children had solitary abscess and the remaining 33 children, multiple abscesses . Of the children with multiple abscesses, 23 had abscesses confined to one lobe of the liver and 10 had involvement of both lobes of the liver . 86 children had culture of liver abscesses done and only 63 (73.2%) yielded positive culture . Staphylococcus aureus and Escherichia coli were the commonest organisms cultured from liver abscesses . 72 cases had drainage of liver abscesses, one of them ended up with hepatic artery ligation . A further 15 cases treated by PTHD survived . Of the remaining 49 cases who had antibiotic therapy, 2 died of septicemia, giving a mortality rate of 1.47% . 36.3% of children with liver abscesses responded to antibiotic therapy . The indication and method for surgical management are discussed . Percutaneous transhepatic drainage (PTHD) of liver abscesses, under the guidance of ultrasonography is found to be safe and effective.

Arerugi, 1989 Jan, 38(1), 36 - 40
{Effect of lobenzarit disodium (CCA) on B cell differentiation to antibody secreting cells}; Imai F et al.; We investigated the effect of the immunomodulator CCA on B cells, using the system of Staphylococcus aureus Cowan 1-stimulated B cells developing into antibody secreting cells in the presence of T cell factors . Results were expressed as the number of plaque-forming cells (PFC), as measured by reverse hemolytic plaque assay . Serially diluted CCA was added to the culture system to evaluate its effect (500-0.005 micrograms/ml) . Different results were obtained by adding CCA at a high dose or a low dose . High-dose CCA showed an inhibitory effect on PFC (p less than 0.01) . On the other hand, low-dose CCA showed an inhibitory effect on high PFC responses of B cells and a stimulatory effect on low responses . Our data suggest that CCA has a direct effect on B cells and that low-dose CCA has immunomodulatory action on normal variations in immune responses.

Ann Med Interne (Paris), 1989, 140(2), 99 - 101
{Septicemia in patients undergoing chronic hemodialysis}; Ben Hamida M et al.; Between March 1982 and March 1986, 11 out of 72 patients (15%) undergoing chronic hemodialysis in a Tunisian hemodialysis unit developed septicaemia . The causative organism was a staphylococcus aureus in 8 patients, and the main portal of entry was the dialysis catheter (6/8) . Septic pulmonary metastasis were observed in 3 cases . REmoval of the dialysis catheter and antibiotic therapy led to recovery in 7 cases . The 4 fatalities were attributed to respiratory failure in 3 cases and septic shock in one case . These results indicate that the prognosis of septicaemia in hemodialysis patients is poor.

Microbios, 1989, 58(235), 83 - 93
Microcalorimetric studies of the effects of platinum group metal complexes on bacterial growth; Bunker JC et al.; A range of platinum group metal complexes (PGMC) were screened for antibacterial activity against Klebsiella aerogenes and Staphylococcus aureus . The effects of the complexes were monitored by changes in the thermal and growth properties of the organisms . Active complexes caused an immediate cessation of power and biomass production which did not recover whilst the complex was present in the medium . Removal of the complex by washing the cells allowed the growth of the small viable proportion of the cells; the majority of the cells had been killed . Changes which occurred in aqueous solutions of active complexes rendered them less bactericidal; this was a possible cause of regrowth observed at sub-bactericidal levels of some complexes . Resistance to active complexes could not be achieved by serial subculture . Three palladium complexes active against K . aerogenes all had similar square planar structures, but in general it was not possible to correlate activity with structure . The unique effects of the PGMCs at low concentrations, combined with the apparent inability of the bacteria to develop resistance to them, makes the complexes a valuable addition to the antibacterial arsenal.

Microbios, 1989, 57(231), 93 - 8
Inhibition of direct haemolytic plaques by antistaphylococcal sera; Carmona Pla MP et al.; An in vitro study was carried out to determine the IgM response to staphylococcal antigens (Staphylococcus aureus, serotype 3, ATCC 12600), at different stages of the growth curve . This was in order to observe the number of inhibited haemolytic plaques when immunosera obtained from blood-letting in rabbits previously immunized with antigenic components were used.

Microbiol Immunol, 1989, 33(1), 81 - 5
A simple method for observation of phagocytosis on bacterial thin-layer; Seki K et al.; A simple method was developed to visually present the phagocytic activity of leukocytes by using adherent Staphylococcus aureus cells and blood applied on a plastic dish . When heparinized blood was applied on thin-layer of heat-killed S . aureus cells on the plastic dish, plaques due to the phagocytic activity of leukocytes were observed with a microscope under a low magnification . Fewer and smaller plaques were observed when plasma-deprived rather than whole blood was used . Some analyses were made in respect to the fundamental conditions required for optimal results . This method was considered to be useful for conveniently evaluating the serum opsonin activities and phagocytic function of leukocytes in various kinds of diseases.

Microbiol Immunol, 1989, 33(3), 155 - 63
Simple method for quantitation of cell-bound protein A on Staphylococcus aureus cells by means of hemagglutination with sheep erythrocytes differentially sensitized with rabbit antibody and its clinical application; Hwang SM et al.; A simple method for quantitation of cell-bound protein A (SpA) on organisms of Staphylococcus aureus was successfully devised by using hemagglutination between staphylococcal organisms and a series of sheep erythrocyte suspensions sensitized with different amounts of anti sheep erythrocyte rabbit antiserum . The validity of the principle and the reproducibility of the method presented here were precisely analyzed and the details of the method are presented . The hemagglutination was quantitatively inhibited both by normal rabbit serum and by soluble SpA . Using the method presented here, 376 strains of S . aureus freshly isolated from clinical materials were subjected to SpA quantitation in cell-bound form . According to the results, there was an unexpected distribution profile in the amounts of cell-bound SpA among the clinical isolates, which showed a two-peak pattern . A possible usefulness of the method presented here in clinical investigations is briefly discussed also.

Diagn Microbiol Infect Dis, 1989 Jan-Feb, 12(1), 73 - 80
Synergistic interaction of cefotaxime and its metabolite desacetylcefotaxime demonstrated by drug-impregnated disks; Stevens DL et al.; Because no simple in vitro test for cefotaxime (CTX) and desacetylcefotaxime (dCTX) synergy exists, and, because no disk susceptibility data is available for dCTX, we investigated the potential of antibiotic impregnated disks as a test for positive in vitro interaction . Results of this study indicate that all strains of Staphylococcus aureus tested demonstrated additive interactions using disks that contained 30 micrograms of dCTX . In contrast, it was necessary to use much smaller concentrations of each cephalosporin to demonstrate additive effects against Gram-negative bacteria . Zones of inhibition were greater for disks containing the combination compared to either CTX or dCTX alone, among all the Gram-negative genera tested; however, these differences were statistically significant only for Klebsiella pneumoniae and Escherichia coli (P less than 0.05) . These results demonstrate that disk susceptibility testing is reliable for the CTX metabolite, dCTX, and suggests that this technique using disks impregnated with CTX and dCTX singly and in combination is a simple method with which to evaluate positive antimicrobial interactions in the clinical laboratory.

Eur J Pediatr, 1989 Jan, 148(4), 318 - 9
Infection with Staphylococcus aureus producing toxic shock syndrome (TSS) toxin-1 without TSS; Nadal D et al.; Relevant findings are reported in an 8-year-old boy with skin infection due to Staphylococcus aureus producing toxic shock syndrome toxin-1 without shock but with an increase in antibody titre against the toxin.

Nippon Geka Gakkai Zasshi, 1989 Jan, 90(1), 5 - 11
{Correlation between coagulase typing and antibiotic susceptibility in methicillin resistant Staphylococcus aureus}; Takesue Y et al.; One hundred and fifty-two strains of Staphylococcus aureus isolated from clinical specimens from 1983 to 1987 were examined in their susceptibility against various antibiotics and in their coagulase typing . The isolation frequency of methicillin resistant strains (MRSA) was 61.6%, and highly methicillin resistant (MIC greater than 100 micrograms/ml) group occupied 81.3% of MRSA in 1987 . From the study of coagulase type, it was found that current epidemic strains in our ward was type II . In MRSA, type IV strains were predominantly isolated until 1984, but after 1986 most of all MRSA strains belonged to type II . According to the coagulase type-classification of MRSA, the isolation frequency of type II was 97.5% of highly methicillin resistant group . Type II strains were more sensitive to MINO and OFLX than type IV, although in the susceptibility to DMPPC type II was less sensitive than type IV . In type IV, the percentage of beta-lactamase-producing strains was 79.3%, and type II, 33.3% with the high significance of statistical difference (p less than 0.01) . In conclusion, a close correlation was suggested between beta-lactamase producing activity and the mechanism of resistance to non-beta-lactam antibiotics as to MINO, OFLX in type IV strains . On the other hand, in type II strains the mechanism of resistance seemed to be simply explained by changes of penicillin-binding-protein (PBP), and there was little concern with beta-lactamase, and other antibiotic-modifying enzymes.

Zentralbl Bakteriol Mikrobiol Hyg {A}, 1989 Jan, 270(3), 337 - 44
Levels of toxic shock syndrome toxin-1 production among Staphylococcus aureus strains and clinical implications; Naidu AS et al.; Among 250 S . aureus clinical isolates, the incidence of TSST-1 production was 18.0% . S . aureus var . hominis strains were predominant (95.6%), producing high levels of toxin in vitro, within the range of 0.6 to 4.3 micrograms/ml and exhibiting crystal violet binding with C/D pattern . No correlation was found between the level of TSST-1 production in vitro and the clinical course . Two (3.4%) of the var . bovis strains produced toxin in amounts less than 0.6 micrograms/ml and did not bind crystal violet . None of the 24 var . canis isolates produced TSST-1 . Fifty five per cent of the isolates of phage group I produced TSST-1 and corresponded to 57.8% of the toxigenic strains . Two of the 250 patients developed toxic shock syndrome.

Rev Infect Dis, 1989 Jan-Feb, 11 Suppl 1, S14 - 21
Epidemiology of toxic shock syndrome in the United States: overview; Broome CV; Studies were undertaken to detect a possible reporting or diagnostic bias that might have affected the results of previous epidemiologic investigations of toxic shock syndrome (TSS) . The methods used in this reassessment of initial findings regarding TSS epidemiology (which were obtained through passive- and active-surveillance systems) included record review and analysis of discharge diagnoses . Results obtained with both the latter methods confirmed the broad outlines of the descriptive epidemiology of TSS . Some recent studies have focused on nonmenstrual cases of TSS and on risk factors for the development of TSS . Key findings include the more frequent isolation from nonmenstrual TSS cases of Staphylococcus aureus strains not producing TSS toxin 1 and a higher case-fatality rate for cases in men and for "non-vaginal" cases in women than for vaginal cases (defined as menstrual, postpartum, vaginal nonmenstrual, and contraceptive cases) . Studies of the relation between tampon use and TSS suggest that both the degree of absorbency and the chemical composition of the tampon are involved; however, the mechanism by which tampons increase the risk of TSS remains unknown . Contraceptive sponges and diaphragms may be linked to an increased risk of nonmenstrual TSS, whereas oral contraceptives may play a protective role; these possibilities require further study.

Rev Infect Dis, 1989 Jan-Feb, 11 Suppl 1, S117 - 23; discussion S123-4
Alteration of proteins reacting with antibody to toxic shock syndrome toxin 1 by endogenous proteases of Staphylococcus aureus; Lawellin DW et al.; Many cases of toxic shock syndrome (TSS) have been associated with Staphylococcus aureus strains that produce in vitro a 22,000-dalton protein called toxic shock syndrome toxin 1 (TSST-1) . TSST-1 has been shown to be linked phenotypically to the production of the type II (thiol) staphylococcal protease; however, some strains clearly associated with TSS do not produce TSST-1 . With the use of a variety of antibodies to TSST-1, representatives of both TSST-1-positive and TSST-1-negative TSS-associated strains were shown to produce proteins of 32,000, 53,000, and 76,000 daltons that cross-reacted with TSST-1 but that did not appear to be TSST-1 aggregates or to be protein A . TSST-1-negative strains were significantly (P less than .05) more likely than TSST-1-producing strains to produce the type I protease and less likely to produce the type II protease . In vitro the type I (serine) protease digests purified TSST-1 . When combinations of type II and type III protease chemical inhibitors were used, the production of the high-molecular-weight cross-reacting proteins during in vitro growth could be selectively enhanced . Since the infected foci (e.g., abscess, vagina) associated with TSS may contain natural protease inhibitors, it is possible that such TSST-1 cross-reacting proteins may be expressed in vivo or that levels of TSST-1 may be increased by an inhibitor of the serine protease.

Arch Virol, 1989, 104(1-2), 63 - 75
A bio-engineered rubella E1 antigen; Terry GM et al.; The major rubella envelope protein, E1, and a segment of it, comprising amino acids 207-353, have been separately expressed as fusion proteins with the IgG binding region of Staphylococcus aureus protein A in Escherichia coli . The proteins carry E1-specific antigenicity recognized by monoclonal antibodies raised against whole virus confirming that correct glycosylation is not required for antigenicity . The use of these bioengineered antigens in immunoassays for diagnosis of rubella infection and for immunization in experimental animals is described.

J Trauma, 1989 Jan, 29(1), 75 - 8
The use of a new assay for detecting antibody to Staphylococcus aureus in severely injured patients; Hershman MJ et al.; Investigation of the antibody response to Staphylococcus aureus infection has been hindered by lack of a simple and specific assay . We report an enzyme-linked immunosorbent assay (ELISA) using a strain of S . aureus devoid of Protein A, a frequent cause of false positive results in ELISAs, and have used this assay to study antibody responses of 23 severely injured patients . This IgM ELISA had a diagnostic sensitivity for major Staphylococcal infection of 70% (seven of ten patients with major infection) and a specificity of 92% (12 of 13 patients without major infection) . Of three patients with major Staphylococcal infections who mounted no IgM response, two died and the other developed severe chronic Staphylococcal infection, and hence prompt initiation of appropriate therapy was necessary . Furthermore, the ability to mount IgM response to Staphylococcal infections appears to contribute to an orchestrated host defense response against this organism.

J Clin Invest, 1989 Jan, 83(1), 243 - 51
Role of endotoxemia in cardiovascular dysfunction and mortality . Escherichia coli and Staphylococcus aureus challenges in a canine model of human septic shock; Natanson C et al.; Using different types of bacteria and a canine model simulating human septic shock, we investigated the role of endotoxin in cardiovascular dysfunction and mortality . Either Escherichia coli (a microorganism with endotoxin) or Staphylococcus aureus (a microorganism without endotoxin) were placed in an intraperitoneal clot in doses of viable or formalin-killed bacteria . Cardiovascular function of conscious animals was studied using simultaneous radionuclide heart scans and thermodilution cardiac outputs . Serial plasma endotoxin levels were measured . S . aureus produced a pattern of reversible cardiovascular dysfunction over 7-10 d that was concordant (P less than 0.01) with that of E . coli . Although this cardiovascular pattern was not altered by formalin killing (S . aureus and E . coli), formalin-killed organisms produced a lower mortality and less myocardial depression (P less than 0.01) . S . aureus, compared to E . coli, produced higher postmortem concentrations of microorganisms and higher mortality (P less than 0.025) . E . coli produced significant endotoxemia (P less than 0.01), though viable organisms (versus nonviable) resulted in higher endotoxin blood concentrations (P less than 0.05) . Significant endotoxemia did not occur with S . aureus . Thus, in the absence of endotoxemia, S . aureus induced the same cardiovascular abnormalities of septic shock as E . coli . These findings indicate that structurally and functionally distinct microorganisms, with or without endotoxin, can activate a common pathway resulting in similar cardiovascular injury and mortality.

Arch Dermatol, 1989 Jan, 125(1), 85 - 7
Staphylococcal scalded skin syndrome mimicking acute graft-vs-host disease in a bone marrow transplant recipient; Goldberg NS et al.; A 33-year-old man with mild acute graft-vs-host disease after an allogeneic bone marrow transplant for chronic myelogenous leukemia developed a necrolytic rash 90 days after transplant . A diagnosis of staphylococcal scalded skin syndrome was made when a skin biopsy specimen revealed a split in the granular layer and phage group 2, type 71 Staphylococcus aureus was cultured from the blood.

Cell Signal, 1989, 1(4), 387 - 93
Staphylococcus aureus alpha-toxin activates phospholipases and induces a Ca2+ influx in PC12 cells; Fink D et al.; Staphylococcal alpha-toxin at subcytotoxic concentrations stimulated phosphatidylinositol turnover and arachidonic acid release in undifferentiated cultures of pheochromocytoma PC12 cells . Stimulation of phospholipase A2 but not C was dependent on extracellular calcium . Addition of staphylococcal alpha-toxin to PC12 cells caused a dose-dependent, biphasic increase in intracellular calcium measured by fura-2 fluorescence technique . Elevation of intracellular Ca2+ content occurred with a time course similar to those observed for stimulation of phospholipase A2 . Alteration of membrane structure and formation of staphylococcal alpha-toxin pores facilitating an influx of Ca2+, represent the probable mechanisms by which phospholipases C and A2 are activated, respectively . These results suggest a possible involvement of Ca2+, phosphoinositides and arachidonic acid metabolites in the pathogenic action of staphylococcus alpha-toxin and caution against the general usage of this toxin as a permeabilizing agent to study stimulus-secretion coupling in secretory cells.

Med Dosw Mikrobiol, 1989, 41(3-4), 176 - 83
{Preliminary studies of the effect of steroid hormones on skin bacteria in vitro}; Basta M et al.; It results from studies on skin hormones that some of steroid hormones reach high concentrations on skin surface mainly during a course of acne vulgaris . Our studies indicate that the activity of hormones on skin bacteria can be multiform . Nineteen hormones of analytical grade of purity were tested . They were derived from different firms of synthesized in the Department of Endocrinology of the Pharmacology Institute of Polish Academy of Sciences in Cracow . Bacterial strains tested in this study were isolated from normal skin flora . The majority of the compounds tested in this study showed different reactivity toward Staphylococcus aureus as well as Staphylococcus epidermidis and anaerobic diphtheroids+ and sometimes aerobic ones.

Eye, 1989, 3 ( Pt 6), 685 - 9
Bacterial contamination of intraocular lenses: the source of the bacteria; Spencer SR et al.; In order to investigate potential sources for the bacterial contamination of intraocular lenses, specimens were taken for culture from a variety of sites . A swab from the lid margin, the operating room air, an intraocular lens which was allowed to rest on the eye, and the sodium hyaluronate were cultured during routine intercapsular lens implant surgery on 31 patients . Positive cultures were obtained from six intraocular lenses . Subtyping by three independent methods showed that isolates from five of these lenses were indistinguishable from bacteria cultured from the air . In the case of the sixth lens a bacterial isolate from the lid margin matched with that from the lens . Staphylococcus epidermidis was cultured from four lenses; Staphylococcus haemolyticus was also isolated from one of these four lenses and the remaining two lenses produced isolates of Staphylococcus aureus.

Acta Vet Scand, 1989, 30(3), 335 - 9
Surface properties of Staphylococcus aureus isolated from caprine mastitis; Jarp J et al.; A total of 53 Staphylococcus aureus strains isolated from caprine mastitis were tested for surface hydrophobicity, surface Protein A (SpA), and binding capacity of fibronectin, fibrinogen and type II collagen . Strong positive correlation was found between surface hydrophobicity and SpA, and between surface hydrophobicity and 125I-fibronectin-binding . Regardless of hydrophobicity, the binding of fibrinogen was moderate and type II collagen binding was low . The results indicate that SpA and fibronectin-binding protein contribute to the high relative surface hydrophobicity of S . aureus associated with caprine mastitis.

Med Arh, 1989, 43(2-3), 107 - 12
{The frequency of isolation of Staphylococcus aureus from the nose and throat of patients with staphyloderma . A short review of pathogenic and nonpathogenic bacteria on the skin}; Mutevelic-Arslanagic N; In the first part of this work, we have given our own experiences of the frequency of isolation of staphylococcus aureus from nase and neck of patients with staphylodermia . Staphylococcus aureus is more often isolated from nose and neck, p less than 0.05 of patients with staphylodermia in comparison with control group . In the other part of this work we have given a review of the most frequent pathogenic and apathogenic bacteria on skin . Particularly we paid attention to the place and role of staphylococcus aureus on the human skin.

Carlsberg Res Commun, 1989, 54(4), 157 - 63
Complete amino acid sequence of alpha-acetolactate decarboxylase from Bacillus brevis; Svendsen I et al.; The complete amino acid sequence of acetolactate decarboxylase (EC 4.1.1.5) from Bacillus brevis has been determined by sequencing of the intact enzyme and of peptides obtained by cleavage with cyanogen bromide, Staphylococcus aureus V8 protease and trypsin, respectively . Determination of the C-terminal part was made by treatment with carboxypeptidases Y and M II . The enzyme has a molecular weight of 29,093 and consists of 260 amino acid residues arranged in a single peptide chain without disulphide bonds.

Acta Microbiol Bulg, 1989, 24, 15 - 20
Protein A in the stable L-forms of Staphylococcus aureus; Ivanova E et al.; The presence of protein A in the stable L-form of S . aureus BM 3041 was proved . Its amino acid composition and electrophoretic characteristics were compared with the parent strain . The location of protein A on the cytoplasmic membranes of the L-form cells, as well as on the extracellular membranes were established by immunoelectron microscopy . In the cells of the parent bacterium its location on the cell wall was observed.

Microbiol Immunol, 1989, 33(12), 981 - 90
Altered virulence of a pleiotropic Staphylococcus aureus mutant with a low producibility of coagulase and other factors in mice; Seki K et al.; The virulence of a pleiotropic Staphylococcus aureus mutant with an extremely low producibility of coagulase and other factors was investigated in mouse . A mutant strain, designated as CL-1, showed the same LD50 and the same intrarenal proliferation as its parental strain, when the mutant organisms were inoculated in mice in high doses . The mutant organisms, however, showed a diminished intrarenal proliferation compared with its parental organisms in low doses . This mutant strain expressed a pleiotropic phenotype such as a concomitant reduction in the producibility of coagulase, alpha-toxin, and Panton-Valentine leucocidin . The total effect due to the reduction in producibility of various factors on the virulence of the mutant strain was investigated with studies on the bacterial resistance to the phagocytic activity of leucocytes . A possible role of coagulase and that of some other staphylococcal exoproteins in the pathogenesis of S . aureus were discussed.

Eye, 1989, 3 ( Pt 2), 190 - 3
Staphylococcal infection and the limbus: study of the cell-mediated immune response; Ficker L et al.; The relationship between enhanced cell-mediated immunity (CMI) to staphylococcal antigens, expressed as delayed hypersensitivity (DH), and the development of catarrhal infiltrates at the limbus in the rabbit has been explored by others . This DH is required for infiltrates to develop in the rabbit cornea when it is exposed to conjunctival inoculation with live Staphylococcus aureus cells . Similar investigations have not been pursued in the human, although St . aureus has been isolated from lids of patients with sterile marginal ulcers . We have tested 69 patients with blepharitis, eleven with and 58 without associated symptomatic marginal keratitis, for DH to killed whole cells of St . aureus and St . epidermidis and protein A; quantitative cultures have also been collected from lids and conjunctivae . Preliminary findings show that nine out of 11 patients with symptomatic marginal keratitis, requiring treatment with steroids, have enhanced DH to St . aureus cell wall antigens . We suggest the hypothesis that this type of marginal keratitis in the human is the result of enhanced CMI at the limbus to St . aureus cell wall antigens.

Folia Histochem Cytobiol, 1989, 27(3), 161 - 7
Effect of Staphylococcus aureus serine proteinase on human lymphocyte impairment; Turyna B et al.; Assessment of lymphocyte surface membrane integrity by means of trypan exclusion test and longer term survival studies based on 3HTdR uptake and Con A stimulation test were used to estimate effect of Staphylococcus aureus serine proteinase on human lymphocytes in vitro . At concentration of proteinase as high as 10 micrograms/ml, the percentage of trypan-stained lymphocytes increase with enzyme level . In contrast, decrease of 3HTdR level and Con A stimulation is already apparent at concentrations of proteinase ten times lower . Autologous serum exerts strong protective effect . Observed impairment is discussed as significant for local response in inflammation.

Acta Microbiol Hung, 1989, 36(1), 19 - 24
Toxic shock syndrome toxin-1 (TSST-1) production in staphylococcal infections and serological implications with patients and healthy controls; Naidu AS et al.; TSST-1 production occurred in 4.6% among Staphylococcus aureus strains isolated from divergent clinical sources in South-West Hungary . Patients suffering from staphylococcal infections, whether or not harbouring TSST-1 positive strains, exhibited low anti TSST-1 titres in their acute phase sera compared to the healthy control population . A S . aureus strain isolated from toxic shock syndrome (TSS) was a high collagen binder and a low fibronectin and fibrinogen binder suggesting the role of connective tissue adhesion in colonization and low invasive property of TSS strains.

Trans R Soc Trop Med Hyg, 1989 Jan-Feb, 83(1), 77 - 9
Tropical pyomyositis associated with Trypanosoma brucei rhodesiense infection in a Europid; Weinberg JR et al.; A 29-year-old European woman became infected with Trypanosoma brucei rhodesiense in the Luangwa valley, Zambia . Six days after the initial presentation of this infection she developed evidence of tropical pyomyositis (TP) . These diseases, both of which are rare in Europids, were satisfactorily treated . The pathogenesis of TP, which is nearly always caused by Staphylococcus aureus, is undetermined . It seems possible that in this case either (i) both infections were introduced simultaneously by a tsetse-fly bite, or (ii) T . b . rhodesiense produced multiple focal necroses in skeletal muscles which acted as niduses for the staphylococcal infections; immunodepression caused by this parasite might also have been important.

Med Dosw Mikrobiol, 1989, 41(2), 81 - 5
{Piglets as a model for studying the activity of staphylococcal shock syndrome toxin (TSST-1)}; Zaleska M et al.; In this study the influence of toxic shock syndrome toxin produced by Staphylococcus aureus on the organism of piglets of Minnesota race is presented . Animals were tested in two groups: conventional and gnotobiotic and they were given the toxin intradermally in two doses 10 and 100 micrograms/kg of body weight . It was shown that piglets are sensitive to the toxin which induced rise of body temperature, various clinical symptoms and in functional changes of several organs as shown by analytical studies . Gnotobiotic animals showed lower susceptibility to the toxin as compared to conventional piglets what suggest a participation of endotoxin in pathologic process.

Leuk Res, 1989, 13(10), 937 - 42
Low IL-1 beta production in leukemic cells from progressive B cell chronic leukemia (B-CLL); Aguilar-Santelises M et al.; In vitro production of IL-1 beta by cells from 32 patients with benign monoclonal lymphocytosis of undetermined significance (MLUS) and B cell chronic lymphocytic leukemia (B-CLL) was investigated . Normal B lymphocytes (2 x 10(6)) secreted approximately 5 ng/ml of IL-1 beta during 24 h and approximately ten times more after stimulation with Staphylococcus aureus, strain Cowan 1 (SAC) . When patients were studied, a loss of IL-1 beta production was found in leukemic cells from progressive disease . Cells from MLUS patients secreted near normal levels of IL-1 beta and responded to SAC stimulation, whereas cells from patients with progressive B-CLL produced no, or little IL-1 beta, and did not respond to SAC . Loss of IL-1 beta production in progressively growing B-CLL may be related to an increased malignant character of these cells . This is discussed in relation to the immunogenicity of the leukemic cells and their capacity to differentiate.

J Arthroplasty, 1989, 4 Suppl, S23 - 32
Treatment of major wound necrosis following total knee arthroplasty; Lian G et al.; Following total knee arthoplasty, seven patients developed significant wound necrosis and dehiscence, requiring wound coverage with soft tissue flaps . Three patients had rheumatoid arthritis, three had degenerative arthritis, and one had osteosarcoma of the distal femur . Five different prostheses were used and the wound problems were discovered on average 21 days after arthroplasty . The average wound size was 6.0 cm2 . Five were infected, four with Staphylococcus epidermidis and one with Staphylococcus aureus . All patients were treated with antibiotics and local debridement for an average of 10 days prior to the flap procedure . Medical gastrocnemius muscle flaps were used in two patients, unipedicle flaps transposed from the lateral thigh in three, and bipedicle flaps shifted from the medial thigh in two . Flaps were done an average of 56 days after arthroplasty, and knee rehabilitation was delayed an average of 76 days after arthroplasty . Patients were followed an average of 48 months after the flap procedure . Six patients had mild or no knee pain and one who remained infected had moderate constant pain . Three of the patients had greater than 90 degrees of knee motion and one had 75 degrees of motion . The remaining three had only 35 degrees of motion, due in part to significant preoperative contractures, infection, local radiation, and chemotherapy . Late infection developed in two patients at 20 and 45 months following the flap procedure . There was one excellent, three good, two fair, and one poor result using the Hospital for Special Surgery knee rating system at final follow-up examination.

Free Radic Res Commun, 1989, 7(3-6), 325 - 33
Staphylococcal arthritis--effects of superoxide dismutase on infected knee joints of rabbits; Linhart WE et al.; The effect of SOD on staphylococcal arthritis has not been successfully evaluated to date . We developed an animal model to investigate the correlation . Using 16 rabbits divided into four groups, we injected two groups with staphylococcus aureus and the other two with NaCl . One group in each was also injected with SOD . The presence of SOD activity in untreated and infected knee joints of rabbits over a period of 72 hours showed no significant difference . TBA-reactive substances (TBARS) measured in joint fluid and plasma did differ in each of the groups, with the highest values found in animals with septic arthritis treated with SOD . This finding corresponded especially with the histological investigation . Joints of infected animals intra-articularly injected with SOD also showed histologically significantly more inflammation, a higher amount of bacteria in the joint cavity, and more distinct joint damage than joints injected only with bacteria . The mechanisms responsible for this SOD effect remain to be determined.

Arch Neurobiol (Madr), 1989, 52 Suppl 1, 149 - 54
{Infective complications of the central nervous system (CNS) in addicts to parenterally administered drugs}; Garces JM; Given the high rate of infection by HIV in the APAD in our country, the description of localized infectious problems in the CNS secondary to drug-addiction should always take into account opportunistic infections or tumours occurring there . The initial clinical evaluation should highlight or rule out the presence of clinical indications (specially polyadenia and oral candidiasis) which suggest a clinical condition of immunodepression . Septic embolization due to bacteremia and fungemia is common among drug-addicts, sometimes causing, although not very often in comparison with other sites, various infectious complications in the CNS (meningitis, cerebral abscess, subdural empyema or epidural abscess), "Staphylococcus aureus" being the micro-organism most frequently involved . From 1977 to 1986 the work-group for the study of infections among drug-addicts has listed 6,481 infections . Disseminated candidiasis (582 cases) and infectious endocarditis (506 cases) were the most frequent types of primary infection . Only 33 cases of infection of the CNS were observed, meningitis being the most frequent (57%) . The usual empirical antibiotic treatment for meningitis is cefotaxime or cefotriaxona plus cloxacillin, and for cerebral abscess we substitute metronidazole for isoxazolic penicillin . Given the sexual habits frequently associated with APAD, neurosyphilis is suspect, since immunodepression secondary to infection by HIV causes changes in its natural behaviour . Tetanus is the most serious and uncommon infectious complication connected with drug-addiction . In our society, up to 1986 only 5 cases had been described . As for paludism, the cases detected in this country have belonged to two small outbreaks: one with four cases in Madrid with "Plasmodium vivax" and the other with three cases in Tortosa (Tarragona) with "P . falciparum".

Acta Derm Venereol, 1989, 69(4), 330 - 2
Acitretin induces an increased adherence of S . aureus to epithelial cells; Lianou P et al.; Recently, synthetic retinoids have been implicated as causing a rise in the incidence of staphylococcal infections in patients orally treated with these compounds for various disorders of keratinization . Since the adherence of bacteria to epithelia is an important early event in the development of bacterial infections, in the present study we investigated the in vitro effects of acitretin on the adherence of Staphylococcus aureus to epithelial cells of the anterior nares of 15 healthy human subjects . It was found that pre-incubation of nasal epithelial cells with acitretin causes a statistically significant (p less than 0.001) increase in the adherence of S . aureus to these cells, as compared to the controls . The growth of S . aureus cultures in the presence of acitretin exerted no effect on the staphylococcal adherence . These results suggest that the oral acitretin-induced increase in S . aureus colonization and in the incidence of cutaneous staphylococcal infections may be related to the enhancement of staphylococcal adherence to epithelia caused by this compound.

Rev Infect Dis, 1989 Jan-Feb, 11 Suppl 1, S83 - 8; discussion S88-9
Toxic shock syndrome toxin 1 is encoded by a variable genetic element; Kreiswirth BN et al.; The primary cause of toxic shock syndrome is toxic shock syndrome toxin 1 (TSST-1), a 22,049-dalton exotoxin . Approximately 20% of Staphylococcus aureus isolates produce TSST-1; the production of this toxin is therefore a variable genetic trait . The TSST-1 gene and its flanking sequences are found on a genetic element that is present in TSST-1-positive isolates and absent in TSST-1-negative isolates . Preliminary sequence data and Southern hybridization experiments with the cloned flanking sequences have provided evidence that the TSST-1 element is 4-7 kilobases in size . Hybridization analysis of whole-cell DNA from two genetically mapped TSST-1-positive strains has demonstrated that the TSST-1 element has at least two chromosomal locations . This finding suggests that the element is mobile . Biotyping of 75 TSST-1-positive isolates showed that the large majority were tryptophan-negative, and Southern hybridization analysis of whole-cell DNA from these isolates revealed a common blotting pattern--an observation suggesting that these strains are clonal.

Biotechnol Ther, 1989-90, 1(4), 335 - 46
Antimicrobial activity of synthetic bactenecin; Gallis B et al.; Bactenecin is an antimicrobial peptide isolated from bovine neutrophils . Bactenecin was synthesized by solid-phase peptide synthesis and renatured to a fully disulfide bonded form . The peptide inhibits the growth of Escherichia coli and Staphylococcus aureus at the same concentration reported for the peptide purified from bovine neutrophils . Bactenecin inhibits the growth of other medically important bacteria and yeast, and it kills the fungus Trichophyton rubrum . Acetylation and amidation of the amino- and carboxy-termini of bactenecin do not change its potency, while replacement of its two cysteine residues with serine decreases the potency.

Zhonghua Zhong Liu Za Zhi, 1989 Jan, 11(1), 57 - 9
{Post-operative pleural infection of lung cancer and its influence on survival}; Cai LF; 1139 patients with primary lung cancers resected from 1954 to 1984 are reported . Twelve of them had pleural infection post-operatively, an infection rate of 1% . The infection caused mainly by Staphylococcus aureus developed on Days 5-23 post-operatively . Most of the patients had fever . Closed drainage was applied for each case . Two of these 12 patients survived for more than 5 years . They had Stage 1 and Stage 2 small cell carcinoma and adenocarcinoma . No significant difference was observed in the 5-year survival rate (33.3% and 30.6%) between the infected group and uninfected group . There was no relation between the post-operative pleural infection and prognosis . With review of related literature, beneficial immunologic response leading to better prognosis might exist in the infected cases, but further observation and discussions are needed.

Mol Microbiol, 1989 Jan, 3(1), 65 - 78
Construction, transfer and properties of a novel temperature-sensitive integrable plasmid for genomic analysis of Staphylococcus aureus; Luchansky JB et al.; As an alternative approach to genetic transfer and analysis, a novel integrable plasmid system was developed that should prove useful for mapping and cloning various genes in Staphylococcus aureus and other Gram-positive bacteria . The use of a restriction-deficient recipient strain and an improved protocol for protoplast plasmid transformation facilitated direct cloning of a recombinant plasmid (pPQ126) in S . aureus NCTC 8325-4 . Plasmid pPQ126 (13.6 kb) is a novel, temperature-sensitive integrable plasmid containing genes encoding resistance to erythromycin and chloramphenicol (from plasmid pTV1ts), and resistance to gentamicin (from transposon Tn4001) . When introduced into an appropriate recipient strain at the permissive temperature (30 degrees C), pPQ126 replicates autonomously . Integration of pPQ126 is directed into homologous chromosomal target sequences (chromosomal insertions of Tn551 or Tn4001) by growing a population of cells containing autonomous pPQ126 in the presence of gentamicin, erythromycin, and chloramphenicol at 39 degrees C (nonpermissive temperature) . Elevated temperature both selects for and maintains pPQ126 as an integrated replicon . Integration of pPQ126 occurs at significantly reduced frequency in a recombination-deficient host, and does not occur in the absence of host chromosomal homology . Integrated pPQ126 excises from the chromosome under permissive conditions (30 degrees C), and excision results in derivatives of pPQ126 that harbour DNA of chromosomal origin.

Proc Natl Acad Sci U S A, 1989 Jan, 86(1), 51 - 5
Enzymatic cleavage of a bacterial genome at a 10-base-pair recognition site; Weil MD et al.; The circular genome of Staphylococcus aureus was cut into two fragments by a simple enzymatic method that cleaves a 10-base-pair site . The recognition sequence, A-T-C-G-mA decreases T-C-G-mA-T, was created by the combined use of the methylase M.Cla I (A-T-C-G-mA-T) and the restriction endonuclease Dpn I (G-mA decreases T-C) . This technique is insensitive to CpG methylation and in human DNA is predicted to produce fragments that, on average, are greater than five million base pairs . The ability to create such long pieces of DNA should facilitate mapping of large, complex chromosomes.

Int J Immunopharmacol, 1989, 11(8), 871 - 7
Effect of dexamethasone on mechanisms responsible for regulation of polyclonal B-cell response; Pryjma J et al.; The effect of dexamethasone (Dex) on the differentiation of pokeweed mitogen (PWM), or Staphylococcus aureus Cowan I (SAC)-stimulated human peripheral blood mononuclear cells (PBMC), into immunoglobulin secreting cells (ISC) was studied with special emphasis on the regulatory role of IL-2 in these systems . Dex, known to reduce endogenous IL-2 production and expression of IL-2 receptors, reduced the proliferation of pokeweed mitogen-activated T-cells, and the proliferation was restored by exogenous recombinant interleukin 2 (rIL-2) . Furthermore, Dex enhanced in PWM and in SAC-stimulated cultures, the number of ISC . Addition of rIL-2 resulted in a further increase of ISC in SAC-stimulated cultures, whereas in PWM-stimulated cultures the enhancing effect of Dex was reversed . When IL-2 receptors were blocked by a monoclonal anti-IL-2 receptor antibody rIL-2 was no longer suppressive . Addition of monocytes to PWM-stimulated cultures resulted in suppression or the number of ISC, which was even more pronounced when monocytes were pretreated with rIL-2 . In contrast to ISC, neither a suppressive effect of rIL-2 nor an enhancing effect of Dex was observed when PWM-stimulated cultures were evaluated for cells with intracytoplasmic immunoglobulin (plasma cells) . From these results we conclude that Dex, by blocking IL-2 production and receptor expression, interferes with IL-2 mediated induction and/or activation of suppressor mechanisms.

Med Dosw Mikrobiol, 1989, 41(2), 86 - 91
{Bacteriophage conversion as a factor modifying the intensity of phagocytosis of Staphylococcus aureus by human leukocytes}; Mlynarczyk G et al.; Staphylococcus aureus NCTC 8325-4 and its eight variants lysogenized with phages responsible for the synthesis of staphylococcal staphylokinase were used for the study . Influence of phage conversion of S . aureus on its interaction with human leucocytes and influence of prophage on strain susceptibility to intracellular killing by human granulocytes without opsonins were evaluated . It was found that lysogenization of the strain with the bacteriophages decreased in each case reactivity of human leucocytes for staphylococcal strain what was expressed by lower bioluminescence values and by lower percentage of intracellular killing of bacterial cells carrying prophage.

Zahn Mund Kieferheilkd Zentralbl, 1989, 77(1), 8 - 11
{Escape of hydrogen peroxide and oxygen from the root canal}; Glockmann E et al.; In vitro investigations demonstrate that hydrogen peroxide, like the oxygen emitted from the root canal, escapes into the area around the root . The amount measurable was dependent on the concentration, the amount of time that the hydrogen peroxide was left in the root canal and the patency of the foramen apicale . The growth of the Staphylococcus aureus (SG 511) was restricted in the area around those roots whose root canal contained a 5% solution of hydrogen peroxide.

Jpn J Antibiot, 1989 Jan, 42(1), 212 - 47
{Evaluation of the efficacy of ceftriaxone in acute suppurative otitis media and acute exacerbation of chronic suppurative otitis media . A comparative study with cefotiam as the control}; Baba S et al.; In order to objectively evaluate the efficacy and the safety of ceftriaxone (CTRX) using once daily administration of 1 g to cases of acute suppurative otitis media and acute exacerbation of chronic suppurative otitis media, a group comparison study by the envelope method was conducted using cefotiam (CTM) as the control drug (2 g twice daily) . The results obtained are summarized as follows . 1 . Clinical efficacies evaluated by the committee were 71% in the CTRX group and 86% in the CTM group for acute suppurative otitis media, and 63% and 60%, respectively, for chronic suppurative otitis media . When all cases were considered both groups evidenced a clinical efficacy of 64%, and no significant difference was observed between the 2 groups . 2 . Clinical efficacies evaluated by the physician in charge were 65% in the CTRX group and 86% in the CTM group for acute suppurative otitis media, and 72% and 60%, respectively, for chronic suppurative otitis media . When all cases were considered efficacies were, respectively, 70% and 64%, showing no significant difference between the 2 groups . 3 . Bacteriological efficacies were 88% in the CTRX group and 86% in the CTM group for acute suppurative otitis media, and 74% and 62%, respectively, for chronic suppurative otitis media . With all cases bacterial eradication rates were, respectively, 76% and 67% . Bacterial eradication rates were always higher for the CTRX group than for the CTM group, but the difference was not significant between the 2 groups . 4 . Against infections caused by Staphylococcus aureus alone, CTRX showed equal clinical and bacteriological efficacies to CTM . 5 . As side effects, dermatitis, vomiting, and malaise were observed in 5 cases (4%) of the CTRX group and 3 cases (3%) of the CTM group . As clinical testing abnormalities, elevations of GOT, GPT, and Al-P, and thrombocytopenia were noted only in 3 cases (5%) of the CTRX group . Furthermore, all of these abnormalities were temporary and of moderate degree or mild, thus the safety of either drug was considered high . 6 . Clinical utilities were 71% in the CTRX group and 86% in the CTM group for acute suppurative otitis media, and 72% and 62%, respectively, for chronic suppurative otitis media . When all cases were included, they were 72% and 66%, respectively, and there was no significant difference between the 2 groups . It is concluded from the above results that CTRX is a highly useful drug with once daily administration of 1 g in the treatment of suppurative otitis media.

Proc Natl Acad Sci U S A, 1989 Jan, 86(2), 699 - 703
Nucleotide sequence of the gene for a fibronectin-binding protein from Staphylococcus aureus: use of this peptide sequence in the synthesis of biologically active peptides; Signas C et al.; Binding of cells of Staphylococcus aureus to fibronectin, which may represent a mechanism of host tissue adherence, involves a fibronectin-receptor protein present on the bacterial surface . Cloning of a gene coding for a staphylococcal fibronectin-binding protein and construction of a fusion protein with fibronectin-binding properties was previously reported from our laboratory . We have now sequenced the gene and deduced a primary sequence of the fibronectin-binding protein . The protein resembles other cell-wall-associated proteins on Gram-positive bacteria in that it (i) appears to be anchored in the cell membrane via its C-terminal end, (ii) contains a proline-rich repeating unit outside the membrane anchor, and (iii) contains a long (36-amino acid) signal sequence at the N terminus . The fibronectin-binding activity has been localized to a domain composed of a 38-amino acid unit repeated completely three times and partially a fourth time; the identity between the three 38-amino acid sequences varies from 42 to 87% . Three synthetic peptides mimicking the structure of each 38-amino acid unit were constructed . All three peptides interacted with fibronectin, as indicated by their ability to inhibit binding of fibronectin to staphylococcal cells, whereas an unrelated 37-amino acid peptide showed no inhibitory activity.

Virology, 1989 Jan, 168(1), 119 - 27
Mapping of a phosphorylation site in the 176R (19 kDa) early region 1B protein of human adenovirus type 5; McGlade CJ et al.; The 176-residue (176R) early region 1B (E1B) protein of human adenovirus type 5 (Ad5) was shown to be phosphorylated at serine in lytically infected KB cells at a level estimated to be about one phosphate group per 28 176R molecules . Through the analysis of peptides generated by cleavage with cyanogen bromide and Staphylococcus aureus V-8 protease the phosphorylation site was mapped to Ser-164 . Using site-directed mutagenesis, a mutant was produced in which the codon for Ser-164 was changed to that of asparagine while leaving the coding sequence for the overlapping 496R protein unchanged . This virus, which replicated well on human KB cells, produced normal levels of 176R, but in an unphosphorylated form . The mutant transformed baby rat kidney cells in cooperation with E1A at an efficiency about one-half that obtained with wt E1B . These data therefore gave little indication that phosphorylation is essential for the function of 176R.

J Immunol, 1989 Jan 1, 142(1), 167 - 72
Expression of IL-1 receptors on human peripheral B cells; Tanaka Y et al.; The expression of IL-1R on human peripheral B cells was analyzed by the binding assay with 125I-labeled human rIL-1 alpha and by the flow cytofluorometry by the use of FITC-conjugated IL-1 alpha . The proliferation and the differentiation of B cells stimulated with Staphylococcus aureus Cowan I (SAC) in the presence of T cell-derived factors were dependent on IL-1 . By the binding experiment with 125I-labeled IL-1 alpha, B cells expressed only few IL-1R without any stimulations . When they were stimulated with SAC, IL-1R on B cells began to increase by only 1 h, reached the maximum level at 6 h . The binding of 125I-labeled IL-1 alpha to B cells was inhibited by the addition of either cold IL-1 alpha or IL-1 beta suggesting that IL-1R on B cells reactive for IL-1 alpha and IL-1 beta were identical . By Scatchard plot analysis, the existence of two classes of IL-1R on B cells was found . A major class of IL-1R (320 molecules/cell) has a lower affinity (Kd = 3.8 x 10(-10) M) and a minor class of IL-1R (70 molecules/cell) has a higher affinity (Kd = 4.4 x 10(-12) M) . When B cells were stimulated with SAC, both lower and higher affinity IL-1R were increased to 1960 molecules/cell and 300 molecules/cell, respectively . Furthermore, IL-1R on B cells were also detected with FITC-conjugated IL-1 alpha by a flow cytofluorometer . Only 3 to 5% of B cells expressed IL-1R without any stimulations . When B cells were stimulated with SAC, IL-1R-positive B cells were increased to 20% . The addition of anti-class II antibodies inhibited B cell proliferation and differentiation induced with SAC, IL-1, and T cell-derived factors . Anti-class II antibodies also inhibited the number of IL-1R on B cells . These results suggest that the expression of IL-1R was induced as the initial stage of B cell activation and that class II Ag play an important role for the expression of IL-1R on B cells.

Rev Elev Med Vet Pays Trop, 1989, 42(3), 383 - 8
Sero-prevalence of agglutinins to Listeria monocytogenes in Nigerian domestic animals; Oni OO et al.; A survey using tube agglutination test was conducted to determine the antibody prevalence to Listeria monocytogenes serotypes 1/2a, 1/2b, 1/2c, 3a and 4b in 1,190 serum samples of 8 animals species from various sources in Kano and Kaduna states of Nigeria . Following absorption with Staphylococcus aureus antigen to remove cross-reacting agglutinins, 52 (68.4 p . 100) of the horse samples were positive . Twenty-six (36.1 p . 100) pig, 52 (20.8 p . 100) cattle, 50 (20.0 p . 100) goat, 20 (20.0 p . 100) dog, serum samples were also positive . Free-ranging chickens had an antibody prevalence of 18 (32.1 p . 100) while those intensively managed had 3 (6.8 p . 100), a difference found to be statistically significant (P less than or equal to 0.01; X2) . Sheep sera collected from Zaria abattoir had a prevalence of 30 (14.7 p . 100) while those from Ahmadu Bello University, Veterinary Teaching Hospital had 6 (13.0 p . 100) prevalence . The prevalence in camel was 4 (4.3 p . 100) . Overall, of the 1,190 serum samples tested, 26 (21.9 p . 100) were sero-positive for L . monocytogenes agglutinins . Each species of animal tested for L . monocytogenes was positive for all five serotypes, except camel which was negative for serotype 3a . Fourty-four (53.0 p . 100) samples were positive at a titre of greater than or equal to 480 for serotypes 1/2a, 60 (58.3 p . 100) for 1/2b, 57 (52.3 p . 100) for 1/2c, 7 (13.7 p . 100) for 3a and 23 (39.0 p . 100) for 4b . It is concluded that L . monocytogenes infection is widespread in domestic animals in Nigeria.

Proteins, 1989, 6(3), 240 - 8
Engineering subtilisin BPN' for site-specific proteolysis; Carter P et al.; A combination of protein engineering and substrate optimization was used to create variants of the serine protease, subtilisin BPN', which efficiently and specifically cleave a designed target sequence in a fusion protein . The broad substrate specificity of wild-type subtilisin BPN' is greatly restricted by substitution of the catalytic histidine-containing of the catalytic histidine 64 with alanine (H64A) so that certain histidine-containing substrates are preferentially hydrolysed (Carter, P., Wells, J.A . Science 237:394-399, 1987) . The catalytic efficiency, (kcat/Km), of this H64A variant was increased almost 20-fold by judicious choice of substrate and by installing three additional mutations which increase the activity of wild-type subtilisin . The most favorable substrate sequence identified was introduced as a linker in a fusion protein between a synthetic IgG binding domain of Staphylococcus aureus protein A and Escherichia coli alkaline phosphatase . The fusion protein (affinity purified on an IgG column) was cleaved by the prototype H64A enzyme and its improved variant, efficiently and exclusively at the target site, to liberate an alkaline phosphatase product of the expected size and N-terminal sequence . Several features of H64A variants of subtilisin make them attractive for site-specific proteolysis of fusion proteins: they have exquisite substrate specificity on the N-terminal side of the cleavage site and yet are broadly specific on the C-terminal side; they can be produced in large quantities and remain highly active even in the presence of detergents, reductants (modest concentrations), protease inhibitors, at high temperatures, or when specifically immobilized on a solid support.

Gynecol Obstet Invest, 1989, 28(4), 191 - 4
Rabbit plasma fibronectin levels associated with Staphylococcus aureus enterotoxin B: an acute-phase reaction; Bawdon RE et al.; Variation in fibronectin (Fn) levels and white blood cell counts (WBC) following staphylococcal enterotoxin B (SEB) or SEB + cryoprecipitate containing Fn challenge was studied in New Zealand white rabbits . Increased plasma Fn levels were observed 2 h after the intravenous injection of SEB and peaked at 48-72 h (from a mean level 194.6 +/- 4.5 micrograms/ml prechallenge Fn level to a 72-hour postchallenge mean level of 407.9 +/- 25.4 micrograms/ml) . Fn levels then decreased over the succeeding 5 days to approximately prechallenge levels . The total WBC count decreased by 88% within 2 h after the SEB injection . A slow increase in circulatory WBC was observed over the next 24 h . SEB caused an increase in plasma Fn levels and decreased WBC counts with lymphopenia that was followed by a normal lymphocyte count within 5 days . These data suggest that an acute-phase reaction was induced by interleukin-1 . Fn prophylaxis provided no change in clinical signs when given at the time of SEB injection.

Gynecol Obstet Invest, 1989, 28(4), 185 - 90
Fibronectin and postpartum infection in rabbits: an animal model; Bawdon RE et al.; A rabbit model was explored in preliminary studies of uterine colonization and infection, following cesarean section (CS), by toxigenic Staphylococcus aureus (Harrisburg) and other bacterial species known to be isolated from traumatized human uteri; effects on host susceptibility of fibronectin prophylaxis were studied . Histological characteristic and plasma fibronectin levels were recorded in double-blinded studies of four groups of rabbits (55 total), all sacrificed 96 h after CS performed at term . Animals which were inoculated without fibronectin prophylaxis, showed stable or slightly elevated plasma fibronectin levels through 96 h . Rabbits inoculated with nontoxigenic S . aureus (Wiley) exhibited stable fibronectin levels and less microscopic evidence of infection . All inoculated animals were colonized, but those that received fibronectin cryoprecipitate intravenously at CS showed less microscopic evidence of infection . Fibronectin levels for rabbits scores as 'seriously infected' were nearly 1.5 times that of controls . This is in contrast to other clinical studies in which chronic infections and poor clinical outcome have been strongly correlated with decreased plasma fibronectin levels . Histochemical studies of antifibronectin binding suggested that less fibronectin is deposited throughout the endometrial stroma of animals that received fibronectin prior to bacterial challenge . An increased polymorphonuclear leukocytic reaction in the stroma near the incisions was observed in animals that received fibronectin prophylaxis, with or without bacterial challenge . This reaction may be characterized as an acute-phase reaction.

Int J Immunopharmacol, 1989, 11(7), 717 - 23
Interferon-alpha and interferon-gamma as regulators of human adult and newborn B-cell function; Punnonen J et al.; The role of interferon-alpha (IFN-alpha) and interferon-gamma (IFN-gamma) in the regulation of B-cell function was studied using highly purified tonsillar B-cells and unfractionated mononuclear cells from healthy adults and newborns . Recombinant IFN-gamma (rIFN-gamma) was found to enhance Staphylococcus aureus Cowan I (SAC)-induced proliferation of purified B-cells . Recombinant IFN-alpha (rIFN-alpha) neither enhanced nor inhibited the function of purified B-cells . However, when unfractionated mononuclear cells were studied, no enhancement in SAC- or pokeweed mitogen (PWM)-induced proliferation was found in response to these IFNs . In fact, the responses in these cultures were mainly inhibitory . The effects were dependent on the mitogen used for activation of the cells and on the source of the cells . The IFNs acted in a more inhibitory way in cultures stimulated with PWM than in those stimulated with SAC . In addition, they acted more suppressively in cord blood mononuclear cells (CBMCs) than in adult peripheral blood mononuclear cells . PWM was found to decrease CD4+/CD8+ T-cell ratio, and cells with suppressor function physiologically dominate among CBMCs . Thus, the effect of these interferons on B-cell function seems to depend on the magnitude of suppressor cell function among the cell population.

Chemotherapy, 1989, 35(4), 273 - 7
An in vitro investigation of the intraphagocytic bioactivity of difloxacin, ciprofloxacin, pefloxacin and fleroxacin; van Rensburg CE et al.; In this study the intraphagocytic bioactivity of difloxacin, ciprofloxacin, pefloxacin and fleroxacin was investigated using human neutrophils and a combination of a radioassay, a colony-counting method and a fluorescence microassay which enables us to differentiate between intracellular bacteriostatic and bactericidal mechanisms . Staphylococcus aureus and Listeria monocytogenes were used as the test intraphagocytic microbial pathogens . It was found that difloxacin, ciprofloxacin and to a lesser extent pefloxacin and fleroxacin possess intracellular bacteriostatic activity for S . aureus and L . monocytogenes.

Nephrol Dial Transplant, 1989, 4(4), 278 - 81
The influence of calcium mupirocin nasal ointment on the incidence of Staphylococcus aureus infections in haemodialysis patients; Boelaert JR et al.; Mupirocin was used in haemodialysis patients in an attempt to eradicate nasal carriage of Staphylococcus aureus and to prevent infection caused by this microorganism . The effectiveness of calcium mupirocin as a 2% nasal ointment OB2 (16 patients for 104 patient-months) was compared to that of placebo (18 patients for 147 patient-months) in a double-blind study . Mupirocin or placebo were applied in both anterior nares thrice daily for 2 weeks and subsequently three times weekly for a total of 9 months . During therapy, S . aureus was recovered from only 6% of the nasal cultures in the mupirocin group compared to 58% in the placebo group (P less than or equal to 0.01) . Only one S . aureus infection was documented in the mupirocin group compared to six in the placebo group (P less than or equal to 0.05) . The S . aureus strain causing the single infection in the mupirocin group was of a different phage type to that of the original nasal strain . In contrast, at least four of the six strains causing infection in the placebo group were of similar phage type to the original nasal strain . All S . aureus isolates remained mupirocin sensitive (MIC less than or equal to 1 mg/l) . In conclusion, mupirocin nasal ointment was effective in eradicating nasal carriage of S . aureus and in preventing S . aureus infections in patients on haemodialysis.

Arch Microbiol, 1989, 151(5), 391 - 8
A similar protein portion for two exoglucanases secreted by Saccharomyces cerevisiae; Ramirez M et al.; Exoglucanase (exo-1,3-beta-D-glucan glycohydrolase, EC 3.2.1.56) activity secreted by Saccharomyces cerevisiae into the culture medium was separated by ion exchange chromatography into two glycoprotein isoenzymes which contributed 10% (exoglucanase I) and 90% (exoglucanase II) towards the total activity . Analysis of the "in vitro" deglycosylated products by polyacrylamide gel electrophoresis under native or denaturing conditions indicated that the protein portions of both exoglucanases exhibited identical mobility, each one consisting of two polypeptides with Mr of 47,000 and 48,000 . The same profile was shown by the exoglucanase secreted in the presence of tunicamycin . Antibodies raised against the protein portion of exoglucanase II did react with both native exoglucanases and their deglycosylated products with a pattern indicative of immunological identity . Digestion of the "in vitro" deglycosylated products of both exoglucanases with Staphylococcus aureus V-8 protease or trypsin generated the same proteolytic fragments in each case . Only exoglucanase II was secreted by protoplasts . These and previously reported results indicate that the protein portions of both isoenzymes may be the product of the same gene (or a family of related genes), and that exoglucanase I is a product of enzyme II, modified by a process occurring beyond the permeability barrier of the cell.

J Pak Med Assoc, 1989 Jan, 39(1), 6 - 9
Methicillin resistant staphylococcus aureus in a teaching hospital of Karachi--a laboratory study; Ashiq B et al.; A search was made for Methicillin Resistant Strains of Staphylococcus Aureus (MRSA) among staph aureus cultures isolated at a teaching hospital in Karachi . Of 100 staphylococcus aureus isolated in 1987-88, 5 were MRSA, four from admitted patients and one from outpatient . These MRSA were resistant to Gentamicin as well as to other antibiotics . The presence of MRSA in the in-patients is a serious problem as it can act as reservoir to cause outbreak of colonisation and infection . No MRSA was isolated from 50 samples studied from Quetta.

J Appl Bacteriol, 1989 Jan, 66(1), 27 - 35
Variation in the behaviour of enterotoxigenic Staphylococcus aureus after heat stress in milk; Batish VK et al.; The survival of several strains of Staphylococcus aureus after heat stress in different menstrua was not logarithmic and F-values were determined to express their resistance to heat . Of the strains tested, Staph . aureus 234 (enterotoxin B) was the most heat resistant and Staph . aureus 790 (enterotoxin E) was the most heat sensitive . Buffalo milk gave the best protection to all the strains of Staph . aureus against heat, followed by cow's milk; phosphate-buffered saline gave the least protection . Soyabean casein digest agar gave maximum recovery of survivors followed by brain heart infusion and Baird-Parker medium . At 50 degrees C there was no marked variation in coagulase production by the surviving strains but at 55 and 62.5 degrees C there was complete loss of coagulase activity . There was a decreased deoxyribonuclease (DNase) production by all the strains of Staph . aureus after heat stress . Heat-treatment at 55 and 62.5 degrees C resulted in loss of enterotoxin production by all the survivors except S6 and 234, the surviving cells of which still produced enterotoxin B after heat treatment at 55 degrees C . Most of the survivors regained lost characteristics such as coagulase, DNase and enterotoxin production after four to five passages through BHI which suggests that subculture of Staph . aureus recovered from heat-processed milk is necessary to avoid false results.

Biochimie, 1989 Jan, 71(1), 117 - 23
Membrane structure and dynamics by 2H- and 31P-NMR . Effects of amphipatic peptidic toxins on phospholipid and biological membranes; Dufourc EJ et al.; The actions of bee venom melittin and delta-lysin from Staphylococcus aureus on membranes have been monitored by solid-state deuterium and phosphorus NMR and shown to differ depending on temperature and on the lipid-to-peptide molar ratio Ri . In the gel phase of phosphatidylcholine model membranes, for lipid-to-peptide ratios Ri greater than 15, melittin induces isotropic lines interpreted as reflecting the presence of small discoidal structures, whereas delta-lysin does not . These small objects are metastable, that is, within a time-scale of hours they return to large lipid bilayers . The kinetics of this process depend on the lecithin chain length . In the fluid phases, at temperatures greater than that of the gel-to-fluid transition Tc, analysis of the quadruplar splittings in terms of chain ordering indicates that both melittin and delta-lysin similarly disorder the membrane . At temperatures above but close to Tc, melittin preferentially orders the center of the bilayer, while delta-lysin promotes ordering throughout the entire bilayer thickness . These effects are interpreted as reflecting different locations of the peptides with respect to the membrane surface . The addition of greater amounts of toxins, Ri = 4, on phosphatidylcholine model membranes induces very small structures irrespective of the temperature in the case of melittin, but only above Tc for delta-lysin . NMR spectral features similar to those characterizing the small fast-tumbling objects with phosphatidylcholine are also observed with egg phosphatidylethanolamine and erythrocyte membranes . The formation of small structures is thus inferred as a general process which reflects membrane supramolecular reorganization.(ABSTRACT TRUNCATED AT 250 WORDS)

Rev Infect Dis, 1989 Jan-Feb, 11 Suppl 1, S75 - 81; discussion S81-2
Analysis of toxic shock syndrome isolates producing staphylococcal enterotoxins B and C1 with use of southern hybridization and immunologic assays; Bohach GA et al.; A study was undertaken to evaluate the production of enterotoxin B (Ent B), EntC1, and toxic shock syndrome toxin 1 (TSST-1) by isolates of Staphylococcus aureus from patients with toxic shock syndrome (TSS) and from a variety of other sources . Levels of toxin in culture supernatants were measured by a quantitative immunodiffusion assay . Most vaginal TSS isolates produced TSST-1, either alone or with EntC1 . However, strains that produced EntB or EntC1 but did not express TSST-1 were commonly isolated from patients with nonmenstrual TSS; EntB was usually produced alone or--rarely--with EntC1 . These results were confirmed by probing DNA from representative isolates with an internal probe of the EntC1 gene (entC1) . Extensive sequence homology between entC1 and entB enabled detection of both genes under conditions of high stringency . The genomic location of entC1 in strains producing both EntC1 and TSST-1 varied little but was dependent on the mammalian host . In contrast, the genomic location of entC1 or entB in strains producing EntC1 or EntB alone was variable . These results suggest that these genes are contained on mobile elements.

Rev Infect Dis, 1989 Jan-Feb, 11 Suppl 1, S305 - 12
Stimulation of T cells and induction of interferon by toxic shock syndrome toxin 1; Micusan VV et al.; Toxic shock syndrome toxin 1 (TSST-1) is a potent immunomodulating substance isolated from Staphylococcus aureus strains associated with toxic shock syndrome (TSS) . Flow cytometric analysis was used to compare scatter changes in several cell-surface phenotype markers on human mononuclear cells exposed in vitro to TSST-1 or to phytohemagglutinin, a lectin with similar effects on the immune response . The results showed differences between PHA and TSST-1 in the appearance of the tested T cell subset markers and of interleukin 2 receptors . In general, the stimulation of mononuclear cells by TSST-1 was slower than that by phytohemagglutinin . TSST-1 induced the production of interferon in cultures of murine spleen cells . By means of inhibition studies with specific antibodies to interferon, the interferon produced was characterized as the gamma type . Human mononuclear cells exposed to the toxin also produced gamma interferon, with levels similar to those induced by staphylococcal enterotoxin A, a known potent interferon inducer . The induction of gamma interferon by TSST-1 may play a role in the immunosuppression caused by TSST-1.

JPEN J Parenter Enteral Nutr, 1989 Jan-Feb, 13(1), 23 - 5
Contamination of enteral nutrition systems during prolonged intermittent use; Grunow JE et al.; Two related studies were done to determine the incidence of bacterial contamination in enteral delivery systems that were used for 15 and 7.5 hr, rinsed after each use, and reused daily in vitro for 7 and 5 days, respectively . In the first study, systems infusing either a premixed formula (Ensure) or a hand-mixed formula (Vivonex) did not show bacterial growth until the 4th day, 1.0-2.0 X 10 colony-forming units per milliliter (CFU/ml) of Staphylococcus epidermidis . Thereafter there was sporadic growth of different organisms but never increasing growth during the 7 days of infusion . In the second study, systems with Ensure were initially contaminated with Staphylococcus aureus and Escherichia coli and reused for 5 days . S . aureus was eliminated by rinsing, but E . coli persisted in the delivery system at concentrations of 10(3)-10(6) CFU/ml . We conclude that clean enteral nutrition systems can be rinsed after short-infusion periods and reused up to 7 days in vitro without significant contamination; however, once a bag has become heavily contaminated some bacteria cannot be eradicated from the system by rinsing.

J Bacteriol, 1989 Jan, 171(1), 222 - 9
Homologous metalloregulatory proteins from both gram-positive and gram-negative bacteria control transcription of mercury resistance operons; Helmann JD et al.; We report the overexpression, purification, and properties of the regulatory protein, MerR, for a chromosomally encoded mercury resistance determinant from Bacillus strain RC607 . This protein is similar in sequence to the metalloregulatory proteins encoded by gram-negative resistance determinants found on transposons Tn21 and Tn501 and to a predicted gene product of a Staphylococcus aureus resistance determinant . In vitro DNA-binding and transcription experiments were used to demonstrate those purified Bacillus MerR protein controls transcription from a promoter-operator site similar in sequence to that found in the transposon resistance determinants . The Bacillus MerR protein bound in vitro to its promoter-operator region in both the presence and absence of mercuric ion and functioned as a negative and positive regulator of transcription . The MerR protein bound less tightly to its operator region (ca . 50- to 100-fold) in the presence of mercuric ion; this reduced affinity was largely accounted for by an increased rate of dissociation of the MerR protein from the DNA . Despite this reduced DNA-binding affinity, genetic and biochemical evidence support a model in which the MerR protein-mercuric ion complex is a positive regulator of operon transcription . Although the Bacillus MerR protein bound only weakly to the heterologous Tn501 operator region, the Tn501 and Tn21 MerR proteins bound with high affinity to the Bacillus promoter-operator region and exhibited negative, but not positive, transcriptional control.

APMIS, 1989 Jan, 97(1), 9 - 13
A comparative study of two lipases from different strains of Staphylococcus aureus; Rollof J et al.; In order to compare lipases from two different Staphylococcus aureus strains (FN 37 and TEN 5), the enzymes from the respective strains were purified using octyl-Sepharose chromatography and characterized with regard to chemical, immunological and enzymatic properties . Differences in the size of the lipases in their native forms necessitated modifications of the purification process, but after purification identical subunits of about 43 kD were found in SDS-PAGE, and both lipases had an apparent molecular weight of 110 kD when subjected to gel chromatography on Sephadex G-200 . Analysis of the amino acid compositions of the lipases showed considerable differences . Serine was the predominant amino acid in the FN 37 lipase, whereas glycine was most abundant in the TEN 5 lipase . Nevertheless, the two lipases were similar when tested with double immune diffusion against an antiserum raised against the TEN 5 lipase, and inactivation of enzyme activities by the antiserum followed the same patterns . Enzymatically, the enzymes were similar with regard to salt inhibition, ion dependency and heat inactivation . The substrate specificities, tested against glyceride substrates, were similar . Thus, the two staphylococcal lipases seem to be similar but not identical.

J Clin Microbiol, 1989 Jan, 27(1), 210 - 2
Ovine-associated Staphylococcus aureus protein with immunochemical similarity to toxic shock syndrome toxin 1; Ho G et al.; A toxic shock syndrome toxin 1 (TSST-1) antibody-binding protein produced by an ovine-associated strain of Staphylococcus aureus was examined . The protein showed total identity to TSST-1 by immunodiffusion analysis . Western blots (immunoblots) of proteins separated by isoelectric focusing revealed that the TSST-1 antibody-binding protein had a pI of 8.6 rather than 7.0, the pI of standard TSST-1.

Autoimmunity, 1989, 3(4), 261 - 9
B cell hyperresponsiveness in Sjögren's syndrome; Yamoaka K et al.; We examined the production of and responsiveness to B cell growth factor (BCGF) by using proliferation assays in patients with Sjogren's syndrome (SS) . We also studied the ability of SS B cells to respond to B cell stimulatory factors (BSF) by measuring the amount of IgG produced in vitro . SS patients showed normal production of and responsiveness to BCGF . They were also demonstrated to show a normal response to Staphylococcus aureus Cowan I (SAC) . In addition, cell surface antigens of B cells in the peripheral blood of SS patients were not altered compared with normal controls . However, SS B cells produced a significant amount of IgG in vitro in response to BSF stimulation with or without anti-IgM antibody . Furthermore, there was a positive correlation between the hyperresponsiveness of B cells to BSF in vitro and serum IgG level in SS . We, therefore, suggest that the enhanced response of B cells to BSF might contribute to polyclonal B cell activation in SS.

G Batteriol Virol Immunol, 1989 Jan-Dec, 82(1-12), 127 - 32
{Evaluation of a rapid agglutination method for the identification of Staphylococcus aureus}; Fucale G et al.; It has been valued a new quick latex method (Staphaurex Wellcome) for the identification of the aureus Staphylococcus . This shown simultaneously the Clumping Factor (CF) and the A protein; the production of the free Coagulase (CL) and the Thermonuclease (TNase), has been utilized as methods of remarking . On 120 bacterial stocks isolated by many biological materials, belonging to the family of the Micrococcaceae, 56 stocks was identified as aureus . St . and 64 as not aureus St . For what concerns the quick Staphaurex method, it has shown a sensitivity of 83.4, with 8 false negatives on 120 cases, while it proved very satisfactory as specification with one false positive.

Perit Dial Int, 1989, 9(3), 197 - 202
Cytotoxic effects of commercial continuous ambulatory peritoneal dialysis (CAPD) fluids and of bacterial exoproducts on human mesothelial cells in vitro; van Bronswijk H et al.; Cultured human mesothelial cells were exposed to peritoneal dialysis fluids, supernatants from cultures of Staphylococcus aureus and S . epidermidis, and antibiotics . Mesothelial cell monolayer cultures were derived from surgically removed omentum . The cytotoxicity of various agents for the cultured mesothelial cells was measured by a 51 Cr-release assay . All brands of fresh peritoneal dialysis fluids induced a more than 50% 51 Cr-release after 18 h . Morphological changes observed included retraction and shrinking of cells, pyknosis of the nuclei and, finally, detachment of cells over an 18-h period . Neutralization of the acid (pH 5.2-5.5) fluids to pH 7.3 did not abolish the cytotoxicity . In contrast, effluent dialysis fluids were not toxic for mesothelial cells; neither was acid (pH 5.5) culture medium nor culture medium with glucose up to 2% . However, higher glucose concentrations induced increasing 51 Cr-release . Furthermore, filter-sterilized supernatants of S . aureus were cytotoxic for mesothelial cell monolayers in 4/7 (57%) strains of S . aureus tested . In contrast, only 4/29 (14%) strains of S . epidermidis produced cytotoxic exoproducts (p = 0.03) . Antibiotics were not found to be cytotoxic, with the possible exception of erythromycin . We conclude that currently available peritoneal dialysis fluids are cytotoxic for mesothelial cells in vitro and that during episodes of peritonitis exoproducts of some bacterial strains may further reduce mesothelial cell viability.

J Invest Surg, 1989, 2(4), 397 - 408
Inhibition by immunoglobulins of Staphylococcus aureus adherence to fibronectin-coated foreign surfaces; Vaudaux PE et al.; Recent data suggest that fibronectin may favor Staphylococcus aureus infection by promoting attachment to either injured tissues or implanted foreign bodies . Using a previously described in vitro assay, we show that promotion of S . aureus adherence by surface-bound fibronectin, adsorbed on polymethylmethacrylate (PMMA) coverslips, is antagonized by antistaphylococcal antibodies present in immunoglobulin G (IgG) purified from human plasma . Among the different organisms tested, the protein A-deficient strain Wood 46 of S . aureus was the most strongly inhibited by purified IgG or whole serum dose-dependently . Bacterial adherence was not influenced by preincubating fibronectin-coated PMMA with either purified IgG or whole serum . However, inhibition of bacterial adherence was directly related to the extent of IgG binding to S . aureus Wood 46 . When F(ab')2 fragments of purified IgG were tested in the adherence assay, they could also reduce the interaction between S . aureus Wood 46 and fibronectin-coated PMMA . Two other staphylococcal strains were also tested in the adherence inhibition assay: Whereas the protein A-rich strain Cowan I of S . aureus was moderately inhibited by purified IgG or whole serum, S . epidermidis KH 11 was not at all inhibited by IgG which bound poorly to the bacterial cells . This study has demonstrated that bacterial coating by humoral factors, and specifically IgG, may influence significantly subsequent adherence of S . aureus to surface-bound fibronectin.

Int Immunol, 1989, 1(3), 310 - 9
The human immunoglobulin C mu-C delta locus: regulation of mu and delta RNA expression during B cell development; Kuziel WA et al.; Whether the immunoglobulin (Ig) heavy chain genes C mu and C delta are expressed singly or in combination, their transcripts undergo differentiation-specific alterations in membrane (M) versus secreted (S) forms as well as in abundance . To better understand this regulation, we have cloned cDNAs for human delta m and delta s to establish the 3' end of the C mu-C delta transcription unit . Steady state mRNA levels and transcription rates were then analyzed in normal and transformed human B cells representing different maturation and activation states . The ratio of micron/microsecond RNA and of delta m/delta s RNA correlated with developmental stage, with a higher ratio at earlier stages . Steady state ratios of total mu/delta RNA paralleled ratios of C mu/C delta nascent transcription, suggesting no major posttranscriptional control for differential expression . However, at all developmental stages, transcription termination occurred downstream of the micron exons, suggesting a strong posttranscriptional regulatory component for production of secreted versus membrane forms of mu RNA . The relative abundance of mature delta S RNA was considerably higher in the human than in the mouse, correlating with the increased levels of circulating IgD in the former species . Stimulation of human splenocytes with mitogens did not increase delta RNA; in fact, splenocytes activated with pokeweed mitogen were nearly devoid of delta RNA, and Staphylococcus aureus Cowan I caused only a minor change.

Acta Microbiol Pol, 1989, 38(3-4), 271 - 83
Phagocytosis of Staphylococcus aureus by somatic cells in dry cow secretion of mammary gland; Smuda PH; Independently of the stage of the dry period, phagocytosis and killing of Staphylococcus aureus by somatic cells isolated from dry cow secretion were significantly higher in a medium of diluted secretion than in 1% serum and in PBS . Intramammary introduction of vaccine from killed Staphylococcus aureus cells caused, in the steady state dry period, the significant increase of phagocytic and bactericidal activity of somatic cells, examined in the medium of diluted secretion of the mammary gland . The isolated secretion deriving both from vaccinated and non-vaccinated cows causes--in the steady state dry period--the disappearance of the significance of differences between the somatic cells coming from various cows . Phagocytosis is highly significantly correlated with the total and intracellular survival of bacteria; similarly, highly significant correlation exists between the total and intracellular survival of S . aureus.

Acta Microbiol Pol, 1989, 38(2), 185 - 8
Influence of flavonoids on combined experimental infections with EMC virus and Staphylococcus aureus in mice; Panasiak W et al.; Activity of elected flavonoids in EMC virus and Staphylococcus aureus infected mice were investigated, to determine the influence on mortality rate . Some of the flavonoids: quercetin, hypoletin, hesperidin, rutin and quercitrin showed significant protective effect given before single as well as combined viral-bacterial infections.

Acta Microbiol Pol, 1989, 38(2), 131 - 41
Plasmid-linked protection of {14C}-glutamate transport and its oxidation against Cd2+ in Staphylococcus aureus; Tynecka Z et al.; The effect of Cd2+ on {14C}-glutamate transport energized by endogenous respiration and on glutamate oxidation was studied in the Cd2+-resistant and -sensitive Staphylococcus aureus strains . The results indicate that these processes are protected against 10 microM Cd2+ in the Cd2+ resistant strain 17810R by the 2H+/Cd2+ antiporter encoded by the cadA determinant located on a penicillinase plasmid p II17810 . Even at 100 microM Cd2+, glutamate oxidation was only partially inhibited in this organism and this inhibition appeared to be reversible . In the plasmidless variant strain 17810S, which lacks the 2H+/Cd2+ antiporter, both {14C}-glutamate transport and its oxidation was blocked by Cd2+ at 10 or 100 microM . In this strain, Cd2+-mediated inhibition of glutamate oxidation was irreversible . Energetics of glutamate transport in both strains was analyzed.

Acta Microbiol Pol, 1989, 38(2), 117 - 29
Effect of Cd2+ on growth of the cadmium-resistant and -sensitive Staphylococcus aureus; Tynecka Z et al.; The effect of Cd2+ on aerobic and anaerobic growth was studied in the Cd2+-resistant Staphylococcus aureus 17810R which harbours the cadA and cadB markers on a penicillinase plasmid pII17810 . Also the effect of Cd2+ on growth of the plasmidless strain 17810S, sensitive to Cd2+ was investigated . The results indicate that under all growth conditions the Cd2+-resistant S . aureus 17810R is protected against Cd2+ toxicity up to 100 microM Cd2+ by the 2H+/Cd2+ antiporter, the product of the cadA gene . Energetics of growth of both strains under various conditions is also discussed.

Lab Delo, 1989, (9), 65 - 6
{A stat-method of determining antibiotic concentration in biological fluids}; Pasternak NA et al.; The rapid method for measuring antibiotic concentrations in biologic substrates, developed by the authors, is based on the suppression of Staphylococcus aureus 209 P test culture dehydrogenase activity . Comparison of the rapid method and the routine agar diffusion test has shown a high correlation of the results, the correlation coefficient r being 0.84-0.93 and its probability 99.9% . The new technique permits getting an answer in 4 hours, is simple, and saves nutrient media.

Dev Genet, 1989, 10(5), 372 - 85
Temporal and tissue-specific expression of myosin heavy chain isoforms in developing and adult avian muscle; Merrifield PA et al.; We have raised monoclonal antibodies (Mabs) to myosin heavy chain isoforms (MHCs) that have specific patterns of temporal expression during the development of quail pectoral muscle and that are expressed in very restricted, tissue-specific patterns in adult birds . We find that an early embryonic, a perinatal, and an adult-specific, fast myosin heavy chain are co-expressed at different levels in the pectoral muscle of 8-12 day quail embryos . The early embryonic MHC disappears from the pectoral muscle at approximately 14 days in ovo, whereas the perinatal MHC persists until 26 days post-hatching . The adult-specific MHC accumulates preferentially and eventually completely replaces the other isoforms . These Mabs cross-react with the homologous isoforms of the chick and detect a similar pattern of MHC expression in the pectoral muscle of developing chicks . Although the early embryonic and perinatal MHC isoforms recognized by our Mabs are expressed in the pectoral muscle only during distinct developmental stages, our Mabs also recognize MHC isoforms present in the heart and extraocular muscle of adult quail . Immunofingerprinting using Staphylococcus aureus protease V8 suggests that the early embryonic and perinatal MHC isoforms that we see are strongly homologous with the adult ventricular and extraocular muscle isoforms, respectively . These observations suggest that at least three distinct MHC isoforms, which are normally expressed in adult muscles, are co-expressed during the early development of the pectoral muscle in birds . In this respect, the pattern of expression of the MHCs recognized by our Mabs in developing, fast muscle is very similar to the patterns described for other muscle contractile proteins.

Eksp Onkol, 1989, 11(2), 54 - 7
{Effect of splenic cells sensitized with staphylococcal enterotoxin A on the metastasis of Lewis lung carcinoma in mice}; Shcheglovitova ON et al.; The effect of intact mouse spleen cells sensitized in vitro with staphylococcus aureus enterotoxin A (SEA) on spreading of mouse Lewis carcinoma was studied . A significant decrease in a number of metastases in the lungs and in the weight of the lungs was observed after multiple intrapulmonary inoculation of spleen cells treated with SEA for 6 hours . The effect was less marked after inoculation of the sensitized cells intraperitoneally or into the femoral muscle of the leg affected with the tumour . After multiple inoculations of the sensitized cells, the spleen cells of the treated animals develop the state of interferon hyporeactivity to SEA but not to PHA or NDV.

Z Naturforsch {C}, 1989 Jan-Feb, 44(1-2), 153 - 60
Localization and orientation of subunit delta of spinach chloroplast ATP-synthase within the CF0CF1 complex . 1 . Distinction of shielded and exposed surfaces of delta on the thylakoid membrane; Berzborn RJ et al.; A new polyclonal antiserum against spinach CF1 subunit delta was produced in rabbits . It decorates only one band at 21 kDa in Western immunoblots of thylakoid proteins and does not react in ELISA with delta-free four subunit CF1(-delta); therefore it is regarded monospecific . The polypeptide used as immunogen had been purified by HPLC . Earlier antisera against CF1 delta cross-react with CF1 subunit beta . The new antiserum 306 contains different antibodies; some can be absorbed with thylakoids, i.e . by delta within the assembled CF0CF1 complex on the membrane, others still react in ELISA with isolated CF1 . The former antibodies agglutinate thylakoids and inhibit PMS cyclic photophosphorylation . Therefore we conclude that part of the surface of CF1 subunit delta is exposed within the quaternary structure of the ATP-synthase complex of photosynthetically active thylakoids, but part of the surface of delta is shielded . Trypsination of isolated delta occurs at several sites, but this protease does not attack delta in situ, nor does aminopeptidase . Staphylococcus aureus protease V8 digests CF1 delta after isolation at residues Asp53, Glu61, Glu95 and Glu106, but has no access to these residues of delta in situ . Thus CF1 subunit delta seems to be shielded within the CF0CF1 complex to a large degree . Direct agglutination of thylakoids by anti delta serum 306 was weak and could be improved tenfold by a Coombs serum (goat anti rabbit gammaglobulin), whereas an anti beta serum agglutinated directly . From this we conclude that delta is not accessible at the top of the enzyme; the exposed part is extending below the large subunits alpha and beta and oriented towards the membrane.

Jpn J Antibiot, 1989 Jan, 42(1), 193 - 9
{Bactericidal action of lomefloxacin a new pyridonecarboxylic acid derivative}; Koikeda S et al.; Lomefloxacin (NY-198) {(+/-)-1-ethyl-6,8-difluoro-1,4-dihydro-7-(3-methyl-1-piperazinyl)-4-oxo -3- quinolinecarboxylic acid hydrochloride} strongly inhibited the growths of not only Gram-negative Escherichia coli but Gram-positive Staphylococcus aureus . In vivo and in vitro experiments showed deoxyribonucleic acid (DNA) synthesis was specifically inhibited by this drug in E . coli.

Biochemistry, 1988 Dec 27, 27(26), 9075 - 80
Thylakoid membrane protein topography: transmembrane orientation of the chloroplast cytochrome b-559 psbE gene product; Tae GS et al.; Protease accessibility and antibody to a COOH-terminal peptide were used as probes for the in situ topography of the Mr 10,000 psbE gene product (alpha subunit) of the chloroplast cytochrome b-559 . Exposure of thylakoid membranes to trypsin or Staphylococcus aureus V8 protease cleaved the alpha subunit to a slightly smaller polypeptide (delta Mr approximately -1000) as detected on Western blots, without loss of reactivity to COOH-terminal antibody . The disappearance of the parent Mr 10,000 polypeptide from thylakoids in the presence of trypsin correlated with the appearance of the smaller polypeptide with delta Mr = -750, the conversion having a half-time of approximately 15 min . Exposure of inside-out vesicles to trypsin resulted in almost complete loss of reactivity to the antibody, showing that the COOH terminus is exposed on the lumenal side of the membrane . Removal of the extrinsic polypeptides of the oxygen-evolving complex resulted in an increase of the accessibility of the alpha subunit to trypsin . These data establish that the alpha subunit of cytochrome b-559 crosses the membrane once, as predicted from its single, 26-residue, hydrophobic domain . The NH2 terminus of the alpha polypeptide is on the stromal side of the membrane, where it is accessible, most likely at Arg-7 or Glu-6/Asp-11, to trypsin or V8 protease, respectively . As a consequence of this orientation, the single histidine residue in the alpha subunit is located on the stromal side of the hydrophobic domain.(ABSTRACT TRUNCATED AT 250 WORDS)

J Biol Chem, 1988 Dec 25, 263(36), 19684 - 9
Limited proteolysis of the nitrate reductase from spinach leaves; Kubo Y et al.; The functional structure of assimilatory NADH-nitrate reductase from spinach leaves was studied by limited proteolysis experiments . After incubation of purified nitrate reductase with trypsin, two stable products of 59 and 45 kDa were observed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis . The fragment of 45 kDa was purified by Blue Sepharose chromatography . NADH-ferricyanide reductase and NADH-cytochrome c reductase activities were associated with this 45-kDa fragment which contains FAD, heme, and NADH binding fragment . After incubation of purified nitrate reductase with Staphylococcus aureus V8 protease, two major peaks were observed by high performance liquid chromatography size exclusion gel filtration . FMNH2-nitrate reductase and reduced methyl viologen-nitrate reductase activities were associated with the first peak of 170 kDa which consists of two noncovalently associated (75-90-kDa) fragments . NADH-ferricyanide reductase activity, however, was associated with the second peak which consisted of FAD and NADH binding sites . Incubation of the 45-kDa fragment with S . aureus V8 protease produced two major fragments of 28 and 14 kDa which contained FAD and heme, respectively . These results indicate that the molybdenum, heme, and FAD components of spinach nitrate reductase are contained in distinct domains which are covalently linked by exposed hinge regions . The molybdenum domain appears to be important in the maintenance of subunit interactions in the enzyme complex.

J Biol Chem, 1988 Dec 25, 263(36), 19804 - 8
Fluorescent labeling of signal-transducing G-proteins . Pertussis toxin-catalyzed etheno-ADP ribosylation of transducin; Hingorani VN et al.; Nicotinamide 1,N6-ethenoadenine dinucleotide (etheno-NAD, epsilon-NAD), a fluorescent analogue of NAD, was able to serve as a substrate for the bacterial toxin-catalyzed epsilon-ADP ribosylation of signal-transducing G-proteins . Pertussis toxin and transducin were used as a model system to characterize this reaction . Similar to ADP ribosylation using NAD as substrate, the epsilon-ADP ribosylation occurs at the carboxyl-terminal 5-kDa tryptic fragment of the T alpha subunit of transducin with the same labeling stoichiometry; however, the rate of labeling is slightly slower . epsilon-NAD competes with NAD as a substrate which suggests that the epsilon-ADP ribosylation occurs at Cys-347 of the T alpha subunit . The biochemical effects of epsilon-ADP ribosylation on transducin are similar to those of ADP ribosylation and include inhibition of the GTPase and {3H}Gpp(NH)p-binding activities . The epsilon-ADP-ribosylated transducin exhibits a fluorescent spectrum which resembles that of epsilon-ADP with an excitation maximum at 292 nm and an emission maximum of 413 nm . Removal of the amino-terminal peptide of epsilon-ADP-ribosylated T alpha with either Staphylococcus aureus V8 protease or trypsin results in a decrease in the emission intensity . This result suggests that the amino- and carboxyl-terminal peptides of the T alpha molecule may interact with each other as suggested previously (Hingorani, V . N., and Ho, Y.-K . (1987) FEBS Lett . 220, 15-22) . epsilon-NAD should prove to be a useful fluorescent substrate for future studies of the ADP ribosylation reaction in biological systems.

FEBS Lett, 1988 Dec 19, 242(1), 157 - 60
N-terminal amino acid sequence of the chromosomal dihydrofolate reductase purified from trimethoprim-resistant Staphylococcus aureus; Hartman PG et al.; The existence of two distinct dihydrofolate reductases (DHFR) in highly trimethoprim-resistant clinical isolates has been unequivocally demonstrated . The enzymes have been characterized with regard to the affinity for substrates and sensitivity to inhibitors . The chromosomal, trimethoprim-sensitive DHFR was purified to homogeneity by a new simple two-step procedure . Its N-terminal amino acid sequence, determined up to the first 35 amino acids, showed 69% homology with the Escherichia coli DHFR.

Cancer, 1988 Dec 15, 62(12), 2628 - 30
Osteomyelitis in pediatric patients with leukemia; Murphy RG et al.; An 11-year retrospective study showed that there were nine patients with osteomyelitis in our population of pediatric patients with leukemia . Six patients had acute lymphoblastic leukemia (ALL) and three had acute nonlymphoblastic leukemia (ANLL) . Seven of nine patients were in remission at the time of diagnosis of osteomyelitis . The delay in diagnosis, which exceeded 2 weeks in seven patients, was typical . Surgical intervention was required for diagnostic or therapeutic purposes in five patients . Staphylococcus aureus was isolated from only three patients . Informative diagnostic studies included nuclear scans that yielded abnormal results in all patients and erythrocyte sedimentation rates (ESR) that were elevated to greater than 70 mm/h in seven patients . The recognition of features that differ from osteomyelitis in the normal population may facilitate earlier diagnosis.

Biochem Biophys Res Commun, 1988 Dec 15, 157(2), 677 - 83
Norepinephrine and GTP-gamma-S increase myofilament Ca2+ sensitivity in alpha-toxin permeabilized arterial smooth muscle; Nishimura J et al.; A new method for preparing permeabilized smooth muscle fibers from rabbit mesenteric artery has been developed using alpha-toxin, a transmembrane pore-making exo-protein produced by Staphylococcus aureus . After alpha-toxin treatment the fibers developed tension as a function of Ca2+ concentration (EC50 = 890 nM) . But they could not contract without added ATP, indicating ATP is permeable . When the sarcoplasmic reticulum was loaded with 5 X 10(-7) M Ca2+ solution, NE induced a transient contraction in 2 mM EGTA 0 M Ca2+ solution and a transient and maintained contraction in 5 X 10(-7) M Ca2+ solution . GTP-gamma-S, a non-hydrolyzable analogue of GTP, substituted for NE in producing these contractile effects . The analysis of the relationship between Ca2+ and maintained tension revealed that NE and GTP-gamma-S cause increases in Ca2+ sensitivity of myofilament shifting the EC50 to 280 nM and 160 nM, respectively . We conclude that NE or GTP-gamma-S causes an increase in myofilament Ca2+ sensitivity and that G protein may be involved in receptor signal transduction system . alpha-Toxin is a useful tool to permeabilize the smooth muscle tissue to ions and small molecules without any damage of receptor and signal transduction system.

J Biol Chem, 1988 Dec 15, 263(35), 18766 - 75
Lipid analysis of the glycoinositol phospholipid membrane anchor of human erythrocyte acetylcholinesterase . Palmitoylation of inositol results in resistance to phosphatidylinositol-specific phospholipase C; Roberts WL et al.; The glycoinositol phospholipid membrane anchor of human erythrocyte acetylcholinesterase (EC 3.1.1.7) contains a novel inositol phospholipid which in this and the accompanying paper (Roberts, W.L., Santikarn, S., Reinhold, V.N., and Rosenberry, T.L . (1988) J . Biol . Chem 263, 18776-18784) is shown to be a plasmanylinositol that is palmitoylated on the inositol ring . The inositol phospholipid was radiolabeled with the photoactivated reagent 3-(trifluoromethyl)-3-(m-{125I} iodophenyl)diazirine and characterized by various chemical and enzymatic cleavage procedures whose products were analyzed by thin layer chromatography and autoradiography or gas chromatography . Acidic methanolysis of human erythrocyte acetylcholinesterase (Ehu AChE) revealed 18:0 and 18:1 alkylglycerols (0.55 and 0.20 mol/mol AChE, respectively) . Acetolysis was shown by TLC to release alkylacylglycerol acetates from Ehu AChE . Analysis by gas chromatography revealed that 83% of the alkylacylglycerol acetates contained an 18:0 or 18:1 1-alkyl group and a 22:4 (n - 6), 22:5 (n - 3), or 22:6 (n - 3) 2-acyl group . The inositol phospholipid is linked to the anchor by a glucosamine in glycosidic linkage, and deamination with nitrous acid cleaved the glycosidic linkage and released the phospholipid . The deamination and acetolysis products from Ehu AChE were purified by high performance liquid chromatography, and fatty acid analysis following acidic methanolysis of the purified products revealed that 2 fatty acid residues were associated with the deamination product and only one with the alkylacylglycerol acetolysis product . The other fatty acid residue was primarily palmitate and was indicated to be in ester linkage to an inositol hydroxyl(s) . This linkage was shown to be responsible for the resistance of the inositol phospholipid to cleavage by Staphylococcus aureus phosphatidylinositol-specific phospholipase . Deacylation of the inositol phospholipid deamination product by treatment with base removed this palmitoyl group and facilitated release of alkyl- and alkylacylglycerol species by phosphatidylinositol-specific phospholipase C with concomitant formation of inositol 1-phosphate . In contrast, digestion of Ehu AChE with a recently reported anchor-specific phospholipase D resulted in release of plasmanic acids from the intact palmitoylated plasmanylinositol.

J Biol Chem, 1988 Dec 5, 263(34), 18318 - 27
Drosophila basement membrane procollagen IV . I . Protein characterization and distribution; Lunstrum GP et al.; A collagen was isolated from Drosophila E85, Schneider line 2L and Kc cell cultures . The purified protein was characterized and antibodies were raised against it . Immunofluorescence microscopy locates this material to the regions of basement membranes of Drosophila embryos, larvae, and adults . The molecules are mostly, or entirely, homotrimers of one polypeptide chain linked by interchain disulfide bonds . The partial amino acid sequences of a cyanogen bromide cleavage product of this chain are identical with a part of the virtual translation product of the Drosophila pro alpha 1(IV) nucleotide sequence that is reported in the accompanying paper . This gene is at Drosophila chromosome location 25C and was identified by the high homology of one part of it with the noncollagenous carboxyl terminus (NC1) of vertebrate type IV basement membrane collagens (Blumberg, B., MacKrell, A . J., Olson, P . F., Kurkinen, M., Monson, J . M., Natzle, J . E., and Fessler, J . H . (1987) J . Biol . Chem . 262, 5947-5950) . In the electron microscope each molecule appears as a thread with a knob at one end, which contains the carboxyl peptide domains . The variation of flexibility of the thread was mapped along its length . Pulse-chase labeling of cell cultures showed that these molecules associate into disulfide-linked dimers and higher oligomers that can be partly separated by velocity sedimentation and are resolved by sodium dodecyl sulfate-agarose gel electrophoresis . Dimers and higher oligomers formed by overlap of the amino ends of molecules were found . Mild pepsin digestion of Drosophila embryos and larvae solubilized the corresponding disulfide-linked collagen molecules, and Staphylococcus aureus V8 protease peptide maps showed the identity of the collagen derived from animals and from cell cultures . Individual, native molecules have a sedimentation coefficient s20,w = 4.1 S, the dichroic spectrum and amino acid composition of a collagen, and a Tm = 31 degrees C . Positive in situ hybridization with a specific probe for this collagen began 6-8 h after egg laying and showed message in the locations of embryos and larvae which reacted with the antibodies . This included some prominent individual cells in the hemolymph.

J Biol Chem, 1988 Dec 5, 263(34), 18152 - 9
Primary structure of the mating pheromone Er-1 of the ciliate Euplotes raikovi; Raffioni S et al.; The complete amino acid sequence of the mating pheromone Er-1 purified from Euplotes raikovi homozygous for mat-1 was determined by automated Edman degradation of the whole protein and peptides generated by cyanogen bromide, trypsin, Staphylococcus aureus V8 protease, and chymotrypsin . The proposed sequence is: Asp-Ala-Cys-Glu-Gln-Ala-Ala-Ile-Gln-Cys-Val-Glu-Ser-Ala-Cys-Glu-Ser-Leu- Cys-Thr-Glu-Gly-Glu-Asp-Arg-Thr-Gly-Cys-Tyr-Met-Tyr-Ile-Tyr-Ser-Asn-Cys- Pro-Pro-Tyr-Val The calculated molecular weight is 4411.0, which is in agreement with the averaged mass of 4410.2 obtained by fission fragment ionization mass spectrometry . Previously reported values of the native molecular weight, determined by gel filtration, have ranged from 9,000 to 12,000 . Thus, the native structure is likely a dimer (or larger aggregate) of identical subunits with the three disulfide bonds present occurring as intrachain links . Secondary structure predictions suggest a helical structure at the amino terminus . A comparison of the Er-1 amino acid sequence with known protein sequences did not reveal any significant similarities.

Biochim Biophys Acta, 1988 Dec 2, 957(3), 399 - 410
The functional glycoprotein CD9 is variably acylated: localization of the variably acylated region to a membrane-associated peptide containing the binding site for the agonistic monoclonal antibody 50H.19; Seehafer JG et al.; Recent studies have shown that {3H}palmitic acid strongly labels both glycosylated forms (gp22 and gp24) of the signal-initiating cell surface glycoprotein CD9 . We performed a two-dimensional limited proteolysis analysis with Staphylococcus aureus V8 proteinase in order to localize the palmitylation sites to final peptides on both glycosylated forms of CD9 . Analysis of {3H}leucine- and {3H}amino acid mixture-labeled gp22 delineated 4 final peptides of 11, 8, 7 and 4 kDa . gp24 produced a similar pattern with the exception that the 11 kDa peptide was replaced by an N-glycosylated 13 kDa peptide . Since all four final peptides (total molecular mass of 30/32 kDa) could not be accommodated by a parent molecule of 22/24 kDa, it is likely that one of the final peptide coexists in two differently modified states . Palmitic acid labeled the 11 kDa/13 kDa final peptides, and the 7 kDa final peptide, with equal intensity, but was not incorporated into the 4 kDa final peptide, demonstrating that fatty acid is ligated in two distinct regions of the molecule . The 8 kDa final peptide was strongly labeled by {3H}palmitic acid, but only weakly by {3H}leucine . We present evidence that this peptide is derived by further acylation of the region defined by the 7 kDa peptide, and that this occurs in only 15% of the molecules . Palmitic acid is turned over faster at these additional sites, indicating that they may be more accessible to membrane transacylases . Proteolysis of CD9 on the intact cell with papain enabled the highly acylated region to be localized to a membrane-associated fragment which contains the binding site for the agonistic monoclonal antibody 50H.19 . The co-localization of a functional domain with a region of variable acylation suggests that acylation events may play a role in the transduction of the signal initiated by interaction of the antibody with CD9.

Int J Food Microbiol, 1988 Dec, 7(3), 185 - 91
Enterotoxigenicity of Staphylococcus intermedius of canine origin; Hirooka EY et al.; Seventy-three staphylococcal strains isolated from pyrodermatitis in dogs were classified as Staphylococcus intermedius (52 strains) or Staphylococcus aureus (21 strains) on the basis of acetoin formation, anaerobic mannitol fermentation, aerobic maltose fermentation, pigmentation, coagulation of human plasma, and reaction on crystal violet agar . Enterotoxin was produced by 13 of the 52 S . intermedius strains and 6 of the S . aureus strains . The highest percentage of enterotoxigenic strains produced enterotoxins C (6 strains), D (7 strains), and E (6 strains) . Four strains produced the toxic shock syndrome toxin-1 . There was little difference in the antibiotic susceptibility between the enterotoxigenic and non-enterotoxigenic strains.

J Antimicrob Chemother, 1988 Dec, 22(6), 873 - 80
In-vitro and in-vivo studies of trimethoprim-sulphamethoxazole against multiple resistant Staphylococcus aureus; Yeldandi V et al.; Methicillin and gentamicin resistant strains of Staphylococcus aureus (MRSA) remains a cause of significant morbidity and mortality . Vancomycin is usually effective against these strains, but toxicity and expense are significant drawbacks . Resistance to the new quinolones has been demonstrated in vitro and during clinical therapeutic trials . Trimethoprim-sulphamethoxazole has proved to be effective in vitro against staphylococcal strains that are resistant to gentamicin, methicillin, and quinolones . As determined by time-kill kinetic studies, trimethoprim-sulphamethoxazole was rapidly bactericidal . Clinical evaluation of trimethoprim-sulphamethoxazole against MRSA in patients with osteomyelitis is under study . We believe that our data support the use of trimethoprim-sulphamethoxazole as a potentially economical and effective alternative for the treatment of infections caused by MRSA.

Eur J Immunol, 1988 Dec, 18(12), 2043 - 7
Glucocorticosteroid-dependent synergy between interleukin 1 and interleukin 6 for human B lymphocyte differentiation; Emilie D et al.; In order to analyze the effects of interleukin (IL) 6 on human in vitro Ig production B lymphocytes were activated by Staphylococcus aureus Cowan strain I (SAC) in the presence of low concentrations of IL2 (1 U/ml) and dexamethasone (10(-7) M) . Previously we showed that this model of B cell response is completely monocyte dependent . We here demonstrate that, under these experimental conditions, IL6 is able to replace monocytes and stimulate Ig production provided IL1 is also present . Dose-effect curves show that these two monokines act synergistically . This synergy is demonstrable only in the presence of dexamethasone, when B lymphocytes are activated (by SAC) and when T cell help (provided by IL2) is present . It results in the production of both IgM and IgG . Both IL1 and IL6 have to be present during the first 48 h of culture to exert an optimal effect . These results show that IL6 may act on early (as well as on late) stages of normal B lymphocyte differentiation . Moreover, glucocorticosteroids potentiate the synergistic effect of IL1 and IL6 on their B lymphocyte target, an effect comparable to that exerted on hepatocytes.

Arthritis Rheum, 1988 Dec, 31(12), 1473 - 80
Prostaglandin E2 modulation of rheumatoid factor synthesis; Alvarellos A et al.; We examined the influence of prostaglandin E2 (PGE2) on the in vitro synthesis of rheumatoid factor (RF) by purified human B and T lymphocytes stimulated with Staphylococcus aureus Cowan 1 or pokeweed mitogen (PWM) . Supernatants were assayed for total IgM and RF . PGE2 at concentrations of 10(-7) M to 10(-9) M significantly inhibited RF and IgM secretion stimulated by S aureus Cowan 1, a cross-linker of B cell surface Ig . The magnitude of inhibition of RF production was significantly greater than that of total IgM at low PGE2 concentrations (P less than 0.05) . In contrast, PWM-stimulated cultures were only minimally inhibited by PGE2 at all concentrations tested . Since cross-linking of surface Ig renders B cells more susceptible to inhibition by PGE2, heat-aggregated IgG (HAIgG) was added to the PWM-stimulated cultures in an attempt to increase the sensitivity of precursors of RF-secreting cells to the inhibitory effects of PGE2 . Addition of HAIgG markedly increased PGE2-mediated inhibition of RF synthesis without significantly affecting IgM production . Inhibition could not be overcome by the addition of soluble T helper cell factors, indicating that PGE2-mediated suppression was not the result of an inhibitory action of T helper cells . When lymphocytes from patients with rheumatoid arthritis were examined, HAIgG was found to be unable to induce sensitivity to PGE2-mediated inhibition of responsiveness . These results suggest that down-regulation of RF synthesis requires both cross-linking of surface Ig and the influence of PGE2 . Abnormalities in this immunoregulatory mechanism may explain the ongoing production of RF in patients with rheumatoid arthritis.

Jpn J Antibiot, 1988 Dec, 41(12), 2035 - 43
{Efficacy of sultamicillin fine granules in pyoderma, particularly in impetigo contagiosa}; Tokuda Y et al.; Sultamicillin fine granules were administered orally to 21 patients with impetigo contagiosa and 1 patient with erysipelas, and its clinical efficacy and safety were assessed . The dosage was 10-30 mg/kg daily . Clinical efficacies of impetigo contagiosa were "excellent" in 20 cases and "fair" in 1 case, with an overall efficacy rate of 95% . The result of treatment in erysipelas was "good" . Out of 19 strains of Staphylococcus aureus isolated from foci, 17 strains had high beta-lactamase activities . In these 17 strains, sensitivity to the drug was 2-32-fold higher than to ampicillin, and in 14 out of these 17 strains 4-8-fold higher sensitivity was noted . As side effects, mild diarrhea was seen in 2 patients, but those recovered spontaneously in 3-4 days without particular treatment.

J Appl Bacteriol, 1988 Dec, 65(6), 491 - 9
Cross protection between an encapsulated strain of Staphylococcus hyicus and encapsulated strains of Staphylococcus aureus; Yoshida K et al.; Strain ST67P of Staphylococcus hyicus grew diffusely in regular serum-soft agar . With the addition of rabbit antisera prepared with Staph . aureus strains, Smith, NS58D or NS41D, capsular type A, B or C, respectively, the organisms converted to compact type growth . Mice immunized with heat-killed vaccine of strain ST67P showed significant resistance against either homologous or heterologous strains, Smith, NS58D and NS41D . Passive protective activities in rabbit antisera prepared with strains Smith, NS58D and NS41D were absorbed out with either homologous cell surface polysaccharide fraction or cell surface polysaccharide fraction extracted from strain ST67P . Well-defined large capsules were observed in ultra-thin sections treated with rabbit antiserum prepared with homologous strain conjugated with ferritin . Also, the capsule surrounded by ferritin granules was shown in ultra-thin sections treated with ferritin conjugated with antisera prepared with those heterologous strains although the capsular size was significantly smaller than those observed by homologous antiserum.

Vet Immunol Immunopathol, 1988 Dec, 20(1), 87 - 93
Effects of Staphylococcus aureus mastitis on bovine mammary gland plasma cell populations and immunoglobin concentrations in milk; Doymaz MZ et al.; Bovine mammary tissue and milk samples were examined to determine effects of chronic Staphylococcus aureus mastitis on the humoral immune response . Parenchymal and teat end tissues from lactating bovine mammary glands were stained immunohistochemically to determine distribution of immunoglobulin (Ig) G1-, IgG2-, IgA-, and IgM-producing plasma cells . Numbers of all Ig-producing plasma cells tended to be higher in tissues from S . aureus infected quarters compared with controls, but most differences were not statistically different . Numbers of IgG1-producing plasma cells at the Furstenberg's rosette area of infected quarters were significantly (P less than 0.05) higher than uninfected quarters . There were no significant differences in concentrations of Ig isotypes in milk from S . aureus infected and uninfected quarters . Data suggest that the antigenic effect of chronic S . aureus infection on the humoral immune response of the bovine mammary gland is minimal . Persistency of S . aureus infection may result, in part, from suboptimal stimulation or immunosuppression of the mammary immune system.

Infect Control Hosp Epidemiol, 1988 Dec, 9(12), 534 - 41
A prospective study of infections in hemodialysis patients: patient hygiene and other risk factors for infection; Kaplowitz LG et al.; We performed a prospective randomized study on 71 patients on chronic outpatient hemodialysis to determine whether a sterile technique was better than a clean technique for preparation of the skin over the vascular access site prior to cannulation . In addition, we wanted to determine overall and site-specific infection rates, microbial etiologies of infection, and risk factors for infection . The overall infection rate was 4.7 infections per 100 dialysis months; the vascular access-site infection rate was 1.3 infections per 100 dialysis months; and the rate for bacteremia was 0.7 cases per 100 dialysis months . Staphylococcus aureus was the most common pathogen, but infections were equally divided between gram-positive cocci and gram-negative bacilli . Advanced age (P = 0.02), a low Karnofsky activity score (P = 0.05), poor hygiene (P = 0.0004) and number of hospitalizations (P = 0.0002) were risk factors for infections in general while only poor hygiene (P = 0.002) was a risk factor for vascular access-site infection . Sterile preparation of the skin over the vascular access site was no more effective at preventing infection than was clean technique (P = 0.80) . Maintenance of good personal hygiene may be one of the most important measures for prevention of infections in hemodialysis patients.

J Dairy Sci, 1988 Dec, 71(12), 3521 - 4
Functional activity of neutrophils from bovine mammary glands infected with Staphylococcus aureus; Fox LK et al.; To test the effect of Staphylococcus aureus infection on mammary neutrophil function, intramammary neutrophils from S . aureus-infected quarters (n = 8), from adjacent uninfected quarters (n = 8) of S . aureus-infected cows, and from quarters (n = 8) of uninfected cows were collected and incubated with S . aureus in vitro . Mean percent neutrophils phagocytizing, number S . aureus per neutrophil, and log10 viable phagocytized S . aureus/ml were: 53.2, 6.4, and 4.72 . Differences in function of neutrophils collected from infected and uninfected quarters were not statistically significant . Results indicate that function of neutrophils from S . aureus-infected quarters is similar to function of neutrophils from uninfected quarters.

Biotechnol Appl Biochem, 1988 Dec, 10(6), 563 - 7
The effect of Staphylococcus aureus enterotoxin A on proliferation of lymphoid and nerve cells; Alakhov VYu et al.; The effects of Staphylococcus aureus enterotoxin A (SEA) on proliferative activities of human peripheral blood lymphocytes, B-lymphoma cells of the Namalva line, and nerve cells of the PC12 line have been studied . It has been shown that SEA affects these cells in identical ways, producing either a mitogenic or an antiproliferative effect . Studies on the dynamics of cellular responses to the action of SEA have demonstrated that the effects of the toxin are mediated by its interaction with different binding sites on the membranes of target cells . It has been established also that the antiproliferative effect of SEA is not associated with changes in 2',5'-oligo(A)-synthetase activity or in the level of interferon secretion.

J Clin Microbiol, 1988 Dec, 26(12), 2505 - 9
Staphylococcus aureus capsular types and antibody response to lung infection in patients with cystic fibrosis; Albus A et al.; Chronic respiratory tract infections caused by Staphylococcus aureus are common in patients with cystic fibrosis (CF) . Recently, it was shown in a few CF patients that S . aureus isolates produce capsular polysaccharides (CPs) . However, it is not known whether this is a common feature and whether an immune response to CPs in CF is detectable . Therefore, we examined 170 S . aureus isolates from CF patients and healthy individuals for production of CP types 5 and 8 by using monoclonal antibodies . We found that CP-producing staphylococcal isolates were randomly distributed among CF patients and healthy carriers . Eighty-five percent of all isolates produced CPs, 77% of which were type 8 . Examination of one sputum sample by an immunofluorescence technique suggested that production of CPs is not an in vitro phenomenon . S . aureus isolates from various sites of a single person often yielded more than one CP type . A random distribution of S . aureus strains with CP type 5 or 8 from the skin and respiratory tracts of patients and from the skin of healthy individuals was found . Antibody response to CP types 5 and 8, measured by enzyme-linked immunosorbent assay, was not elevated in CF patients with chronic S . aureus lung infection in comparison with healthy carriers . On the contrary, in CF patients the ratios of antibodies to CP 8 were significantly lower (P less than 0.005; alpha = 0.025) . The ratios of antibodies to CP types did not change when monitored longitudinally over several months . This study suggests that the production of CPs is a universal property of S . aureus and that infected CF patients do not have elevated ratios of antibodies to these antigens.

Foot Ankle, 1988 Dec, 9(3), 101 - 6
Treatment of diabetic foot infections: Wagner classification, therapy, and outcome; Calhoun JH et al.; A total of 850 cases of septic diabetic foot infections were reviewed in 355 patients . Age, sex, other chronic diseases, site, etiology, Wagner grade, treatment, and results were analyzed . One third of the patients were in their sixth decade of life . There were 180 women and 175 men . Chronic diseases included hypertension, congestive heart disease, and renal failure . Staphylococcus aureus was the most common bacteria . Treatment was considered to meet protocol standards if Wagner's algorithms and infectious disease principles were followed . Thirty-nine Wagner grade 0 infections were seen: only one was not treated appropriately . Eighty-eight percent treated per protocol healed and the one not treated appropriately failed . In grade 1, 79% of the 154 evaluable patients were treated appropriately with a 86% success rate versus a 53% success rate for those not treated per protocol . Of three quarters of the 64 patients with grade 2 infections treated according to protocol, 73% healed . One of those in grade 2 who was not treated according to protocol healed . In grade 3, 64% of the 251 patients were treated per protocol with a 79% success rate versus a 12% success rate for those who were not treated per protocol . Most of the 189 patients with grade 4 cases were treated according to protocol with 88% success; the 20 not treated per protocol had a 15% success rate . Thirty of the 32 grade 5 patients were treated per protocol and all but one healed . Protocol therapy had a statistically significant effect by chi 2 test in the treatment of all groups.

Br J Haematol, 1988 Dec, 70(4), 495 - 500
Effect of iron on polymorphonuclear granulocyte phagocytic capacity: role of oxidation state and effect of ascorbic acid; Hoepelman IM et al.; It has been shown that iron (III) impairs the function of polymorphonuclear granulocytes (PMN) . We have studied the effect of iron (II), on the membrane function of PMN, by assessing the uptake of radiolabelled Staphylococcus aureus by these cells . Iron (II), significantly impaired PMN phagocytic function . Addition of ascorbic acid reduced uptake further . Ferrous ascorbate, molar ratio 1:20, impaired phagocytic capacity of PMN significantly at iron concentrations as low as 1-10 microM . The toxic effect of iron (II) was not observed when desferrioxamine or transferrin was present in the incubation medium . The oxygen-free radical scavengers thiourea, mannitol and catalase prevented toxicity mediated by ferrous ammoniumsulphate but not by ferrous ascorbate (molar ratio of 1:20) . Although high concentrations of ascorbic acid inhibited the generation of .OH and also the formation of the DMPO-.OH adduct by zymosan stimulated PMN, toxicity of iron increased . Iron (II) impaired the uptake of S . aureus by PMN of a patient with chronic granulomatous disease while iron (III) did not . Iron mediated impairment of PMN function is not only a result of the generation of toxic oxygen metabolites but also of direct interaction of iron (II) or an iron (II)-oxygen intermediate with molecules of the cell membrane.

Ann Ophthalmol, 1988 Dec, 20(12), 468 - 9
An unusual case of floppy eyelid syndrome; Fleishman JA et al.; A 51-year-old obese man with a history of nocturnal upper eyelid eversion secondary to floppy eyelids presented with a Staphylococcus aureus corneal ulcer . No papillary conjunctivitis was present . The corneal ulcer was treated successfully . This case demonstrates the importance of ruling out spontaneous nocturnal lid eversion as a cause for unexplained ocular surface pathology and emphasizes the fact that the floppy eyelid syndrome may present in the absence of classic papillary conjunctivitis.

Am J Surg, 1988 Dec, 156(6), 524 - 8
Morbidity and mortality of an endemic pathogen: methicillin-resistant Staphylococcus aureus; Hunt JL et al.; Over an 8-year period, two epidemics of methicillin-resistant Staphylococcus aureus (MRSA) occurred in a burn unit . Sources of sepsis were the burn wound and lung . Fourteen percent of the patients colonized with MRSA became bacteremic . The mean postburn day of bacteremia was 19 and the mortality rate was 5 percent . MRSA was introduced to the burn unit when a patient was transferred from another unit, on readmission of a previously infected patient, or heavy burn census when MRSA was epidemic in the hospital . Although the morbidity rate associated with MRSA infections was high, the mortality rate was low . Gram-negative sepsis has continued to be more lethal.

J Med Microbiol, 1988 Dec, 27(4), 263 - 70
Production and characterisation of monoclonal antibodies specific for staphylococcal enterotoxin B; Lin YS et al.; We have generated monoclonal antibodies (MABs) to staphylococcal enterotoxin B (SEB) in BALB/c mice . Five out of 20 clones which produce anti-SEB MABs have been characterised . Among them, three produce IgG1/kappa, one produces IgM/lambda, and one apparently produces both IgG1/lambda and IgM/lambda MABs . The anti-SEB titres of ascites fluids range from 3200 to greater than 819200 by ELISA . All of the MABs analysed thus far neutralise the mitogenic response of BALB/c splenocytes to a suboptimal dose of SEB . Also, the induction of suppressor cells by SEB in vitro is reversed by pre-incubating SEB with these MABs . Limited digestion with chymotrypsin, trypsin or Staphylococcus aureus V8 protease yields peptide fragments which have been tested by Western-blot analysis . MABs 1FD7 and 2GD9 are specific for the carboxy-terminal end of SEB, and have a similar, but not identical, binding epitope . MABs 2DA3 and 2HA10 bind to intact SEB but not to cleaved products, and are probably specific for antigenic determinants altered by the cleavage or by the denaturing conditions of the electrophoresis, or by both.

Ann Thorac Surg, 1988 Dec, 46(6), 615 - 8
Successful treatment of empyema thoracis with polymethylmethacrylate antibiotic-impregnated beads in the guinea pig; Mavroudis C et al.; Two hundred nine Duncan-Harley guinea pigs had intrathoracic inoculation with 10(8) Staphylococcus aureus, accompanied by blood and umbilical tape . One hundred fifty-two animals were excluded because of clinical recovery, early death, or complications related to intrathoracic polymethylmethacrylate (PMMA) bead placement . The remaining 57 animals had clinical signs of empyema thoracis and were the subjects of this study . Group I animals (N = 24) served as the controls and had no therapy . Group II animals (N = 14) were treated by intrathoracic placement of placebo PMMA beads . Group III animals (N = 19) were treated by intrathoracic placement of tobramycin sulfate-impregnated PMMA beads . There were no differences between the groups in pleural reaction or pneumonia scores . These findings demonstrate a similar host response to the established infection . Group III, however, had a higher sterilization rate than Groups I and II (p less than 0.05), a finding underlining the therapeutic effect of tobramycin-treated PMMA beads . We conclude that intrathoracic local antimicrobial therapy with slow-release tobramycin-impregnated PMMA beads may enhance empyema treatment by increasing the rate of local sterilization . More experiments are necessary to assess the efficacy of this potentially important therapeutic arm for the treatment of thoracic empyema.

Ann Surg, 1988 Dec, 208(6), 783 - 7
Oxygen as an isolated variable influences resistance to infection; Jonsson K et al.; The relative importance of oxygen in relation to resistance to infection was assessed in 24 mongrel dogs . Rectus abdominis musculocutaneous (MC) and corresponding cutaneous random pattern (RP) flaps based at the level of the xiphoid were elevated, replaced, and sutured . Immediately after the surgical procedure, 0.1 ml saline containing 10(8) Staphylococcus aureus/ml was injected intradermally into six fields of each flap . After the operation, the dogs were caged in controlled environments with different oxygen concentrations at 12%, 21%, and 45% for 3 days . After 24, 48, and 72 hours, lesion size was measured . Different lesion size was noted between the hypoxic and the hyperoxic groups in the MC flaps from the first day on and in the RP flaps from the second day on (p less than 0.05) . Resistance to infection with S . aureus is oxygen dependent, particularly when tissue PO2 is below 40 mmHg.

J Bacteriol, 1988 Dec, 170(12), 5709 - 17
An enhancer of DNA replication; Gennaro ML et al.; cmp, a nucleotide sequence element in the plasmid pT181 of Staphylococcus aureus, acts as an enhancer of DNA replication . When cmp is present on an unrelated vector along with the pT181 origin of replication, it increases the ability of the linked pT181 origin to compete with a coresident pT181 plasmid for the initiator protein RepC . cmp is contained within a 156-base-pair segment, and its deletion from pT181 reduces by twofold the frequency of plasmid replication under derepressed conditions . The enhancer sequence contains a locus of DNA bending, and enhancer activity decreases with distance from the replication origin.

Biochimie, 1988 Dec, 70(12), 1791 - 7
Kinetics of the diacetyl and 2,3-pentanedione reduction by diacetyl reductase (alpha-diketone reductase (NAD)) from Staphylococcus aureus; Gonzalez J et al.; The kinetic mechanism of diacetyl and 2,3-pentanedione reduction by diacetyl reductase from Staphylococcus aureus was investigated . The shape of the primary double reciprocal plots, the product inhibition pattern, and the features of the inhibition by a substrate analogue (acetone) show that diacetyl is reduced via an Ordered Bi-Bi mechanism, and 2,3-pentanedione by an Ordered Bi-Bi or Theorell-Chance mechanism . NADH is the leading substrate in both reactions . Affinity constants for the coenzyme and the substrates and inhibition constants for NAD, acetoin, and acetone were also calculated . This enzyme has a high affinity for NADH; Km (31-50 microM) and Ks (20-27 microM) for this compound are around one-tenth of the NADH intracellular concentration . Therefore, it must operate in vivo saturated with the coenzyme . This condition is not adequate to play the role, formerly proposed for diacetyl reductases, of regulating the equilibrium between oxidized and reduced forms of pyridine-nucleotides.

Arch Surg, 1988 Dec, 123(12), 1465 - 9
Immunomodulation by altered gastrointestinal tract flora . The effects of orally administered, killed Staphylococcus epidermidis, Candida, and Pseudomonas on systemic immune responses; Marshall JC et al.; We investigated the role of microbial products from the gastrointestinal tract in the pathogenesis of immunosuppression in critical illness . Rats were gavaged daily for three weeks with killed Staphylococcus epidermidis, Candida, and Pseudomonas--organisms frequently isolated from the upper gastrointestinal tract of the critically ill patient; nonspecific immunity was measured by the delayed-type hypersensitivity (DTH) response . Gavage with either Pseudomonas or Candida resulted in significant suppression of DTH responses, while gavage with S epidermidis produced modest enhancement of DTH . Animals given Pseudomonas demonstrated impaired ability to localize a subcutaneous Staphylococcus aureus challenge and decreased IgM but enhanced anti-Pseudomonas IgA and IgG responses after intraperitoneal immunization . Pulmonary bacterial clearance at seven days was normal . These findings suggest that gut colonization with Candida or Pseudomonas may contribute to impairment of cell-mediated immunity in the critically ill patient.

J Immunol, 1988 Dec 1, 141(11), 3785 - 90
A T cell receptor-associated GTP-binding protein triggers T cell receptor-mediated granule exocytosis in cytotoxic T lymphocytes; Schrezenmeier H et al.; To study the subcellular events occurring after T cell activation we used cloned human CTL permeabilized with alpha-toxin of Staphylococcus aureus . This method of permeabilization leads to stable transmembrane channels that permit the introduction of small molecules into the cell but preserves the cellular structures and macromolecular contents of the CTL . We used the exocytosis of CTL-specific serine esterases as a marker of T cell activation . The TCR-activated exocytosis is functioning in such permeabilized CTL . Introduction of the membrane impermeable guanosine nucleotide-binding protein (G-protein) activating GTP-analog GTP gamma S into CTL triggers exocytosis if Ca2+ is present . For optimal exocytosis ATP is required . The G-protein inactivating GDP-analog GDP beta S inhibited exocytosis triggered via the TCR-CD3 complex but not that triggered by activating the protein kinase C . If the protein kinase C was depleted in CTL by overnight incubation with phorbolester, the response to GTP-gamma S was reduced by more than 50% . These experiments demonstrate the presence of a G-protein involved in TCR-mediated CTL triggering . In the sequence of signaling steps this G-protein is localized after TCR-triggering but before the formation of the protein kinase C-activating phosphoinositol breakdown product diacylglycerol in the sequence of signaling steps.

Jpn J Antibiot, 1988 Dec, 41(12), 1822 - 40
{Experimental and clinical studies of flomoxef in the field of obstetrics and gynecology . Representative Committee Members of the Research Team for Infections in the Field of Obstetrics and Gynecology}; Matsuda S et al.; Flomoxef (FMOX) has a broad antibacterial spectrum against Gram-positive and Gram-negative bacteria; especially its potent antibacterial activity against Staphylococcus aureus is a significant advantage that may not be found with other cephem compounds . In our determination of its antibacterial potency against various clinical isolates obtained from clinical materials (amniotic fluid, intrauterine secretions, exudates of the pelvic dead space) of patients with various infections, we obtained results representing specific features of this drug . From the results, the drug may be expected to produce an excellent effect in the treatment of various infections . Our study on drug concentrations in body fluids and genital tissues demonstrated a good transfer of this drug into various tissues; in every tissue examined, the drug administered by the usual method in the usual dose yielded a concentration exceeding MIC for principal pathogens, thus promising a good clinical response . Indeed a high clinical efficacy rate of 90.1% (good to very good responses) was obtained in a clinical trial involving 222 cases . Administration of the drug in 2 g quantity daily produced a high response rate of 92.8% . It was especially noteworthy that a good response was obtained in 30 of 32 cases (93.8%) in which other cephem compounds had failed . In evaluation of the bacteriological effect, furthermore, the drug showed an excellent rate of bacterial elimination . In conclusion, this drug is expected to be greatly useful in the light of its good transfer into genital tissues and its strong antibacterial activities against Gram-positive cocci, Gram-negative bacteria and anaerobes as well as against multiple bacterial infections predominating among women with genital infections.

J Clin Microbiol, 1988 Dec, 26(12), 2672 - 4
Effects of blood and different media on the production of toxic shock syndrome toxin 1 by Staphylococcus aureus in the tampon sac method; Reiser RF et al.; The use of three different agar concentrations in the tampon sac method resulted in slightly higher fluid uptake by the tampons when a 0.5% agar concentration was used . However, there was essentially no difference in the total amount of toxin produced . The largest amount of toxic shock syndrome toxin 1 was produced with brain heart infusion agar, followed closely by 3% NZ-amine NAK-1% yeast extract medium . The addition of plasma and serum to the inoculum resulted in increases (62 and 73%, respectively) in toxin production . The addition of whole blood to the inoculum had a variable effect on toxin production, with an increase in the amount of toxin produced with some tampons and not with others . Over fivefold differences in the amount of toxin produced were obtained when duplicate experiments were done on successive days, whereas the differences were less than twofold for experiments done on the same day . This was related to the effect of small changes in the parameters on toxic shock syndrome toxin 1 production.

Artif Organs, 1988 Dec, 12(6), 484 - 90
Protein A immunoadsorption therapy in HIV-related immune thrombocytopenia: a preliminary report; Bertram JH et al.; Nine homosexual patients with immune thrombocytopenia were treated with autologous plasma that had been perfused over silica-immobilized Staphylococcus aureus protein A (SpA) . Pretreatment platelet counts ranged from 10,000 to 98,000 cells/mm3 (mean: 54,000 cells/mm3) . Six patients responded to therapy . Platelets increased by a mean of 95,000 cells/mm3 (p less than 0.007) and reached normal levels (greater than 150,000 cells/mm3) in four patients . Increased platelet counts are presently sustained in these four individuals after 5 months of follow-up . Increases in platelet counts significantly correlated with decreases in platelet-associated IgG (PAIgG), platelet-directed IgG (PDIgG), and immune complexes (CIC) . PAIgG and PDIgG declined by a mean of 67% (p less than 0.003) and 58% (p less than 0.007), respectively . CIC decreased by a mean of 37% (p = 0.02) . Complement was concomitantly activated in all four examined patients . C3a and C5a increased 23-fold and 2.6-fold, respectively, while total hemolytic complement decreased by 50% . Activated complement components and removal of CIC and IgG thus may contribute to the platelet-enhancing activity of SpA immunoadsorption therapy.

Am Rev Respir Dis, 1988 Dec, 138(6), 1439 - 43
Endotoxin-induced suppression of pulmonary antibacterial defenses against Staphylococcus aureus; Harris SE et al.; In order to evaluate the effect of endotoxin on lung host defenses, Sprague-Dawley rats were intravenously injected with either placebo or 5 mg/kg of Escherichia coli lipopolysaccharide B . Two hours after treatment, animals were challenged with Staphylococcus aureus by either low dose aerosol inhalation or high dose intratracheal instillation of the bacteria into the lungs . Quantitative lung bacteriologic examination and bronchoalveolar lavage (BAL) for total and differential cell counts were performed immediately (zero hour) and at 4 h after bacterial challenge . Lung phagocytic defenses against aerosolized S . aureus challenges are provided solely by the alveolar macrophage (AM) in the absence of inflammation . In aerosol-challenged control rats, 20.6 +/- 2.0% of the initial deposited bacterial challenge remained viable in the lung at 4 h . Animals pretreated with endotoxin, however, showed a significant decrease in pulmonary bactericidal activity (31.3 +/- 3.4% bacteria remaining at 4 h), indicating a defect in alveolar macrophage (AM) function . Further assessment of the bactericidal oxidative metabolism of endotoxin-treated AM by luminol-enhanced chemiluminescence indicated an increased production of free radical oxygen species when compared with control nontreated cells in both the unstimulated (66 +/- 4 versus 38 +/- 7 x 10(3) cpm in control) and stimulated (250.5 +/- 17.1 versus 147.1 +/- 6.2 x 10(3) cpm in control) states . Total and differential cell counts in both control and endotoxin-treated aerosol-challenged rats were similar.(ABSTRACT TRUNCATED AT 250 WORDS)

Arch Neurol, 1988 Dec, 45(12), 1331 - 7
Epidural spinal infection in intravenous drug abusers; Koppel BS et al.; The incidence of spinal epidural abscess and disk space infection appears to be rising in intravenous drug-using patients . We report 18 cases seen over three years in two municipal hospitals . Staphylococcus aureus was the most common infective agent, but two patients had Mycobacterium tuberculosis infection . Computed tomography of the spine facilitates diagnosis . Early treatment improves outcome but host factors play a role in recovery.

Ann Emerg Med, 1988 Dec, 17(12), 1321 - 30
Controversies in antibiotic choices for bite wounds; Callaham M; In the last decade much progress has been made in our understanding of animal bites . Two major areas of remaining controversy are discussed . Recent study of human bites has shown that the early literature depicting all human bites as having an extraordinarily high infection and complication rate was biased by its emphasis on human bites of the hand that presented late with infection already present . These bites, the so-called closed-fist injuries (CFI), do indeed have a poor prognosis, but it may be as much due to their location and initial neglect as to the source of the injury . Human bites elsewhere do not seem to have any higher risk than animal bites, which have an infection rate of about 10% . Human bites of the face, lips, and ears are at very low risk for infection (less than 3%) if properly treated . The CFI should be identified early and aggressively irrigated and debrided (if possible) . CFI wounds can be treated on an outpatient basis if uninfected and less than 24 hours old . The economical outpatient antibiotics of choice for CFI are penicillin plus dicloxacillin; the former is needed to cover Eikenella corrodens and the latter to cover Staphylococcus aureus, both common in these wounds . Diabetics with hand infection frequently have Gram-negative infection and may warrant parenteral aminoglycosides . Second- and third-generation cephalosporins are very effective but should be reserved for special situations due to their expense . Prophylactic antibiotics are not indicated for typical bite wounds, which are low risk . The choice of antibiotic (when needed) in other bite wounds is a matter of confusion, because the only scientific data available are in vitro sensitivities, which are a very poor and crude reflection of the clinical reality . Antibiotic effectiveness in vivo is dependent on a complex summation of absorption, tissue levels (not just serum levels), host immune defenses, and the interrelationships between bacterial species present . For dog bite wounds, dicloxacillin and cephalexin are both good choices because they cover most of the broad spectrum of infecting pathogens; dicloxacillin is significantly cheaper.(ABSTRACT TRUNCATED AT 400 WORDS)

Arch Surg, 1988 Dec, 123(12), 1477 - 81
Early burn wound excision and skin grafting postburn trauma restores in vivo neutrophil delivery to inflammatory lesions; Tchervenkov JI et al.; This study assessed the effect of early vs delayed postburn wound excision and skin grafting on the in vivo neutrophil delivery to a delayed-type hypersensitivity (DTH) reaction and a bacterial skin lesion (BSL) . Male Lewis rats were presensitized to keyhole-limpet hemocyanin . Group 1 comprised sham controls . Groups 2 through 4 were given a 30% 3 degrees scald burn, but the burn wounds were excised, and skin was grafted on days 1, 3, and 7, respectively, after the burn . Group 5 comprised burn controls . Twelve days after burn trauma, all rats were injected at different intervals (during a 24-hour period) with a trio of intradermal injections of keyhole-limpet hemocyanin, Staphylococcus aureus 502A, and saline at different sites . In vivo neutrophil delivery to these dermal lesions was determined by injecting indium in 111 oxyquinoline-labeled neutrophils isolated from similarly treated groups of rats . Neutrophil delivery to DTH and BSL lesions was restored to normal by excision and skin grafting of the burn wound one day after burn trauma . Waiting three days after burn trauma to excise and skin graft the wound partially, but not completely, restored the in vivo neutrophil delivery to DTH and BSL lesions . Waiting one week to excise and skin graft a burn wound resulted in no improvement in neutrophil delivery to DTH and BSL dermal lesions . It was concluded that burn wound excision and skin grafting immediately after burn trauma restored in vivo neutrophil delivery to a BSL and DTH dermal lesion . This may, in part, explain the beneficial effect of early aggressive burn wound debridement in patients with burn injuries.

J Antimicrob Chemother, 1988 Dec, 22(6), 923 - 33
An in-vitro evaluation of the cellular uptake and intraphagocytic bioactivity of clarithromycin (A-56268, TE-031), a new macrolide antimicrobial agent; Anderson R et al.; Erythromycin base and its 6-0-methyl derivative clarithromycin were actively accumulated 7.3 +/- 1.2-fold and 9.2 +/- 2-fold respectively by human neutrophils in vitro . The intraphagocytic bioactivities of the antimicrobial agents were investigated using the combination of a radioassay, colony counting method and a fluorescence microassay which facilitates the distinction between intracellular bacteriostatic and bactericidal mechanisms . Staphylococcus aureus, Listeria monocytogenes and Legionella micdadei were used as the test intraphagocytic microbial pathogens . Both agents were found to possess intracellular bioactivity for all three species of bacteria with clarithromycin being consistently more active than erythromycin . Under the assay conditions used both agents were bacteriostatic (intracellularly) for S . aureus and Leg . micdadei and bactericidal for List . monocytogenes . Clarithromycin is clearly a potent intraphagocytic antibiotic and potentially superior in this respect to erythromycin.

Scand J Immunol, 1988 Dec, 28(6), 791 - 9
Phenotypical and functional characterization of the idiotype-positive blood B cells in multiple myeloma; Bloem AC et al.; In this study the idiotype-positive B cells of one patient with smouldering multiple myeloma (IgG kappa) and of one patient with multiple myeloma (IgG lambda) were analysed phenotypically and functionally . As regards the expression of B cell-associated differentiation antigens and size distribution, the idiotype-positive B cells resembled normal IgG-bearing blood B cells . In functional studies the lymphocytes were cultured in vitro with Staphylococcus aureus, pokeweed mitogen, T-cell factors, or combinations of these . After culture, proliferation and differentiation of the idiotype-positive B cells were measured by autoradiography, an idiotype-specific ELISA, and a spot ELISA . The results show that idiotype-positive B cells of both patients are able to proliferate after stimulation in vitro . In contrast to their normal counterparts, however, almost no increase in the amount of secreted idiotype IgG could be induced . This suggests that the idiotype-positive blood B cells have lost some of their ability to respond to exogenous stimuli.

Arthritis Rheum, 1988 Dec, 31(12), 1481 - 91
Changes in B lymphocyte function in rheumatoid arthritis and lupus nephritis after total lymphoid irradiation; Solovera JJ et al.; Patients with rheumatoid arthritis and patients with lupus nephritis underwent total lymphoid irradiation (TLI) . Peripheral blood mononuclear cells from these patients showed a decrease in pokeweed mitogen-induced immunoglobulin secretion after TLI . A defect in non-T cells contributed to the decreased response . No defect in the response of non-T cells to T cell-independent activators of B cells, Staphylococcus aureus Cowan 1 and Epstein-Barr virus, was observed after TLI . We conclude that TLI induces a selective deficit in B cells, which respond to T cell-dependent mitogen stimulation.

Proc Natl Acad Sci U S A, 1988 Dec, 85(23), 8983 - 7
Characterization of a cDNA clone encoding the calmodulin-binding domain of mouse brain calcineurin; Kincaid RL et al.; A cDNA clone corresponding to a portion of the catalytic subunit of calmodulin (CaM)-dependent phosphoprotein phosphatase (calcineurin) was isolated from a murine brain library by expression vector immunoscreening . A beta-galactosidase fusion protein that reacted on Western blots with anti-calcineurin antibodies and biotinylated CaM was purified in preparative amounts using CaM-Sepharose affinity chromatography . Partial digestion of the hybrid protein with Staphylococcus aureus V-8 protease produced several immunoreactive peptides that appeared identical to fragments generated from authentic brain calcineurin . The 1111-base-pair (bp) EcoRI insert contained an open reading frame encoding a protein of 35 kDa followed by a 190-bp 3' noncoding region; seven peptides obtained by partial amino acid sequencing of the bovine brain enzyme were found in the deduced sequence . A domain approximately 12 kDa from the carboxyl terminus was deduced to be the CaM-binding site based on consensus structural features and a sequence of seven amino acids highly related to smooth muscle myosin light-chain kinase . Two regions with identity to protein phosphatases 1 and 2A were found in the amino half of the cloned sequence; however, the intervening sequence contained apparent insertions, suggesting splicing of subdomains . Thus, the structure of calcineurin is chimeric, consisting of conserved catalytic elements and a regulatory CaM-binding domain.

Proc Natl Acad Sci U S A, 1988 Dec, 85(24), 9704 - 8
Identification, cloning, and characterization of an immune activation gene; Lipes MA et al.; We have identified an immune activation gene, denoted Act-2, by differential hybridization screening of an activated T-cell library . The gene is induced rapidly after T-cell activation with phytohemagglutinin, B-cell activation with Staphylococcus aureus Cowan I, and monocyte activation with lipopolysaccharide . We have isolated a cDNA containing the full-length coding region . The deduced amino acid sequence predicts an open reading frame of 92 amino acids, including a very hydrophobic N terminus, which by weight matrix score is predicted to be a signal peptide . Using a baculovirus expression system, we have shown that this gene encodes a secreted product . It is therefore possible that Act-2 represents a newly discovered cytokine.

Biochem Biophys Res Commun, 1988 Nov 30, 157(1), 364 - 72
Characterization of a recombinant murine interleukin-6: assignment of disulfide bonds; Simpson RJ et al.; Murine interleukin 6 (mIL-6) has been synthesized as a fusion protein using a lac operon inducible plasmid in Escherichia coli . The first 8 amino acids are from the N-terminus of bacterial beta-galactosidase and the last 175 amino acids are from residue number 12 to the end of native mIL-6 . This fusion protein is equipotent with the native molecule in the hybridoma growth factor assay and has comparable receptor binding characteristics . The two disulfide bridges in mIL-6 have been identified by Staphylococcus aureus V8 protease peptide mapping and Edman degradation of cystine-containing peptides . It has been shown that there are disulfide bonds between Cys46-Cys52 and Cys75-Cys85.

Biochemistry, 1988 Nov 29, 27(24), 8741 - 51
Photolabeling of membrane-bound Torpedo nicotinic acetylcholine receptor with the hydrophobic probe 3-trifluoromethyl-3-(m-{125I}iodophenyl)diazirine; White BH et al.; The hydrophobic, photoactivatable probe 3-trifluoromethyl-3-(m-{125I}iodophenyl)diazirine ({125I}TID) was used to label acetylcholine receptor rich membranes purified from Torpedo californica electric organ . All four subunits of the acetylcholine receptor (AChR) were found to incorporate label, with the gamma-subunit incorporating approximately 4 times as much as each of the other subunits . Carbamylcholine, an agonist, and histrionicotoxin, a noncompetitive antagonist, both strongly inhibited labeling of all AChR subunits in a specific and dose-dependent manner . In contrast, the competitive antagonist alpha-bungarotoxin and the noncompetitive antagonist phencyclidine had only modest effects on {125I}TID labeling of the AChR . The regions of the AChR alpha-subunit that incorporate {125I}TID were mapped by Staphylococcus aureus V8 protease digestion . The carbamylcholine-sensitive site of labeling was localized to a 20-kDa V8 cleavage fragment that begins at Ser-173 and is of sufficient length to contain the three hydrophobic regions M1, M2, and M3 . A 10-kDa fragment beginning at Asn-339 and containing the hydrophobic region M4 also incorporated {125I}TID but in a carbamylcholine-insensitive manner . Two further cleavage fragments, which together span about one-third of the alpha-subunit amino terminus, incorporated no detectable {125I}TID . The mapping results place constraints on suggested models of AChR subunit topology.

Biochem J, 1988 Nov 15, 256(1), 103 - 8
Glycosylphosphatidylinositol is involved in the membrane attachment of proteins in granules of chromaffin cells; Fouchier F et al.; Incubation at 37 degrees C or treatment of granule membranes of chromaffin cells with Staphylococcus aureus phosphatidylinositol-specific phospholipase C converted from an amphiphilic to a hydrophilic form two proteins with molecular masses of 82 and 68 kDa respectively . Their release is time- and enzyme-concentration-dependent . We showed that they were immunoreactive with an anti-(cross-reacting determinant) antibody known to be revealed only after removal of a diacylglycerol anchor . Furthermore, the action of HNO2 suggests the presence of a non-acetylated glucosamine residue in the determinant . This is one of the first reports suggesting that a glycosylphosphatidylinositol anchor might exist in membranes other than the plasma membrane . We showed that the 68 kDa protein is probably not the subunit of dopamine (3,4-dihydroxyphenethylamine) beta-hydroxylase, an enzyme present in granules in both soluble and membrane-associated forms.

J Biol Chem, 1988 Nov 15, 263(32), 16906 - 10
Sequence of the human erythrocyte phosphoglycerate mutase by microsequencer and mass spectrometry; Blouquit Y et al.; We have previously reported the isolation in pure form of the human erythrocyte phosphoglycerate mutase isozyme B . We now report the sequence of the whole protein and the identification of its N-terminal blocking group . The protein tryptic peptides of phosphoglycerate mutase isozyme B were isolated by high performance liquid chromatography and their sequence determined by microsequencing . The sequence and the nature of the blocking group of the N-terminal tryptic peptide was shown to be N-acetyl-Ala-Ala-Tyr-Lys by mass spectrometry . Overlaps of the tryptic peptides were obtained by studying the V8 Staphylococcus aureus protease peptides of the aminoethylated phosphoglycerate mutase isozyme B either by microsequencing or by mass spectrometry . The procedure used allowed us to obtain the sequence on a very small amount of material and in a short period of time . Our data agree well with those derived from the cDNA nucleotide sequence described by Sakoda et al . (Sakoda, S., Shanske, S., DiMauro, S., and Schon, E . A . (1988) J . Biol . Chem . 263, 16899-16905) . In addition, our data directly indicate that the initiation codon does not introduce a methionine as N-terminal amino acid and allowed the identification of the acetyl N-terminal group.

Vet Rec, 1988 Nov 12, 123(20), 515 - 7
Antibiotic sensitivity of bovine staphylococcal and coliform mastitis isolates over four years; Mackie DP et al.; Eight hundred and forty-eight strains of Staphylococcus aureus and coliforms isolated from milk samples taken from cows with clinical or subclinical mastitis were tested for their sensitivity to a range of antibiotics, comparing strains isolated in 1984, 1985, 1986 and 1987 . The only increase in the proportion of resistant strains occurred with coliforms resistant to ampicillin and neomycin.

J Chromatogr, 1988 Nov 11, 454, 157 - 67
On-line high-performance liquid chromatography-post-column reaction-capillary gas chromatography analysis of lipids in biological samples; Raglione TV et al.; The versatility of on-line liquid chromatography and gas chromatography is further expanded by the addition of on-line derivatization . The on-line fractionation, derivatization and separation system is applied to the characterization of lipids in biological samples . Separation of the triglycerides from the phospholipids was accomplished on a narrowbore (2.0 mm I.D.) 5 microns silica column . The entire triglyceride fraction was transferred to a heated fixed bed reactor for esterification of the fatty acid constituents . Transfer of the derivatized zones to the gas chromatograph was accomplished by the use of a retention gap . Application of the system to the separation and characterization of Staphylococcus aureus is presented.

J Biol Chem, 1988 Nov 5, 263(31), 15880 - 7
Inactivation of lincosaminide antibiotics in Staphylococcus . Identification of lincosaminide O-nucleotidyltransferases and comparison of the corresponding resistance genes; Brisson-Noel A et al.; Resistance to lincomycin by inactivation has been detected in numerous clinical isolates of Staphylococcus; in crude extracts of Staphylococcus haemolyticus BM4610 and Staphylococcus aureus BM4611, inactivation of lincomycin and clindamycin requires the presence of a nucleoside 5'-triphosphate (ATP, GTP, CTP, or UTP) as nucleotidyl donor and Mg2+ as cofactor . The biochemical mechanism of lincosaminide inactivation was elucidated by determination of the structure of inactivated lincomycin and clindamycin by physicochemical techniques, including UV absorption spectrophotometry, 31P and 1H nuclear magnetic resonance, and periodate oxidation . In the two strains, inactivation of lincomycin gave rise to lincomycin 3-(5'-adenylate), whereas clindamycin was inactivated through its conversion to clindamycin 4-(5'-adenylate) . The gene linA' encoding the 3-lincomycin, 4-clindamycin O-nucleotidyltransferase in S . aureus BM4611 has been sequenced and displays 93% homology with the gene linA encoding the 3-lincomycin, 4-clindamycin O-nucleotidyltransferase found in S . haemolyticus BM4610 . The two enzymes are 161 amino acids long and differ by 14 amino acid substitutions.

J Exp Med, 1988 Nov 1, 168(5), 1539 - 51
Production of tumor necrosis factor/cachectin by human B cell lines and tonsillar B cells; Sung SS et al.; The production of TNF/cachectin by human B cell lines and tonsillar B cells was examined . Of the 15 B cell lines examined, 9 cell lines synthesize TNF mRNA constitutively . PMA stimulated most cell lines to accumulate increased amounts of TNF . SeD, 8866P, 32al, RPMI 1788, and four bone marrow-derived EBV-transformed cell lines accumulated high levels of TNF mRNA when stimulated by PMA . TNF production by these cell lines was examined . RPMI 1788 and WIH8 produced little TNF constitutively, but synthesized 5-7 ng/ml TNF when stimulated by PMA . A pre-B cell line, Nalm-6, did not synthesize any detectable amount of TNF mRNA, even with PMA stimulation . Tonsillar B cells could also be stimulated to produce TNF . PMA or Staphylococcus aureus Cowan I strain (SAC) alone stimulated some TNF mRNA accumulation, whereas B cell growth factor (BCGF) or anti-mu did not . This accumulation was synergistically elevated by the combinations of PMA and SAC, or PMA and anti-mu . BCGF increased PMA-, SAC-, PMA plus SAC-, or PMA plus anti-mu-induced TNF mRNA accumulations about twofold . The accumulation of TNF mRNA in tonsillar B cells stimulated by PMA plus SAC was between 32 and 48 h, the same peak interval as the accumulation of TNF and IL-2 mRNA in tonsillar T cells . This is in contrast to PMA or PMA plus A23187-stimulated RPMI 1788 cells in which TNF mRNA accumulation was maximal at 1-2 h . TNF activities found in tonsillar B cell supernatants correlated with the TNF mRNA levels in the cells . However, more TNF activity was found on the second-day than the third-day supernatants, indicating active TNF uptake by the B cells . Cyclosporin A (CsA) inhibited SAC and anti-mu responses in B cells in much the same way as the anti-CD3 responses in T cells . SAC-, PMA plus SAC-, and PMA plus anti-mu-stimulated, but not PMA-stimulated, increases in TNF mRNA accumulations in tonsillar B cells were inhibited by CsA . TNF production seems to increase in parallel with B cell proliferation, but the relationship of these two functions needs to be further examined.

J Immunol, 1988 Nov 1, 141(9), 3043 - 9
Production of B cell-stimulating factors by B cells in patients with systemic lupus erythematosus; Tanaka Y et al.; The production of B cell-stimulating factors (BSF) by B cells in patients with systemic lupus erythematosus (SLE) was studied in vitro . B cells from SLE patients markedly proliferated and differentiated into Ig-producing cells by in vitro culture without any stimulation . The culture supernatant of these B cells contained BSF activity that stimulated Staphylococcus aureus Cowan I-treated normal B cells to proliferate and differentiate into Ig-producing cells . By a Percoll gradient density centrifugation, BSF-producing cells were enriched in the higher density fraction, but were reduced in the lower density fraction . The BSF also stimulated the proliferation and the differentiation of SLE B cells . By a Percoll gradient density centrifugation, SLE B cells responsive to the BSF were enriched in the higher density fraction, but were reduced in the lower density fraction . The Mr of the BSF was estimated as about 18,000 Da by Sephacryl S-200 column chromatography . The BSF fraction did not possess IL-2 and IFN activity, but possessed IL-1 activity, which stimulated murine thymocyte proliferative responses . The BSF activity was partially, but not completely, absorbed by an anti-IL-1 alpha antibody . Furthermore, the BSF possessed IL-4 activity, which induced not only the proliferative responses of normal B cells stimulated with B cell mitogens, but also the expression of low affinity Fc epsilon R/CD23 on normal B cells . The BSF also possessed IL-6 activity, which induced the proliferative responses of IL-6-dependent hybridoma cells, MH-60 BSF2 . Moreover, human rIL-1, rIL-4, and rIL-6 stimulated SLE B cells . These results suggest that SLE B cells spontaneously produce the BSF such as IL-1 alpha, IL-4, and IL-6 and express their receptors on their surface, and the interaction between the BSF and their receptors stimulates SLE B cells to spontaneously proliferate and differentiate into Ig-producing cells as an autocrine mechanism.

Ann Inst Pasteur Microbiol, 1988 Nov-Dec, 139(6), 655 - 64
Effect of growth on surface charge and hydrophobicity of Staphylococcus aureus; Beck G et al.; Modifications in the surface charge and hydrophobicity of Staphylococcus aureus Oxford during growth were studied by analysing electrophoretic mobility and adherence to hydrocarbons (hexadecane), respectively . Bacterial concentration had no effect upon the measurements . Both surface charge and hydrophobicity varied during the exponential phase of growth (1 to 4 h): surface charge decreased significantly (p less than 0.001), while hydrophobicity increased (p less than 0.001) . In the stationary phase (4 to 9 h), the surface charge increased significantly (p less than 0.001), whereas hydrophobicity showed no change . Cationized ferritin decreased the surface charge and had no effect on hydrophobicity . These results suggest that in S . aureus, different structures could be responsible for their surface charge and hydrophobic properties.

Diagn Microbiol Infect Dis, 1988 Nov, 11(3), 177 - 80
Commercially available technique for rapid laboratory detection of methicillin resistance among Staphylococcus aureus; Piper J et al.; The MRS test is a commercially available test for detection of methicillin resistance among Staphylococcus aureus (MRSA), which was compared to standard techniques for efficacy and speed . Among 119 S . aureus strains tested (71 resistant to methicillin), it detected 90% of MRSA strains in 4 hr . The MRS test may have a role as a rapid screening test for MRSA in selected situations.

Allergol Immunopathol (Madr), 1988 Nov-Dec, 16(6), 407 - 11
The ability of granulocytes of bone marrow tissue and vascular pools in atopic patients to absorb latex particles and Staphylococcus aureus; Matusiewicz R et al.; In 68 patients with atopic asthma the ability of the granulocytes from the marrow tissue and vascular pools to engulf neutral latex particles and Staphylococcus aureus bacteria was studied . In patients with atopic asthma at the time of a lull in the disease, an impairment of engulfing of latex particles and S . aureus by the granulocytes in the marrow and vascular pools was demonstrated . The ability of absorbing bacteria and latex by the granulocytes in the tissue pool was similar in the group of patients and in the control group . At the time of asthmatic dyspnoea the ability of engulfing of S . aureus and latex by marrow pool granulocytes was rising steeply, while vascular and tissue pool granulocytes showed an even greater impairment of this ability than at the time of absence of dyspnoea.

J Dairy Res, 1988 Nov, 55(4), 513 - 9
Oxygen concentration in milk of healthy and mastitic cows and implications of low oxygen tension for the killing of Staphylococcus aureus by bovine neutrophils; Mayer SJ et al.; The partial pressure of O2 in milk from normal cows and from cows with mastitis was measured and the concentrations of O2 calculated . Oxygen levels of milk from normal cows were similar to those in venous plasma, but inflammation of the mammary gland led to a dramatic drop in O2 concentration to less than 10% of control values . Intracellular survival of Staphylococcus aureus strain M60 in bovine neutrophils was greater under anaerobic than aerobic conditions . The implications of low O2 concentrations in milk from infected mammary glands for the bactericidal activity of bovine neutrophils is discussed.

J Biochem (Tokyo), 1988 Nov, 104(5), 700 - 5
Complete amino acid sequence of ferredoxin from Peridinium bipes (Dinophyceae); Uchida A et al.; The amino acid sequence of the major ferredoxin component isolated from a dinoflagellate, Peridinium bipes, was completely determined . Staphylococcus aureus V8 proteolytic, tryptic and chymotryptic peptides of Cm-ferredoxin were prepared and sequenced . The sequence was Phe-Lys-Val-Thr-Leu-Asp-Thr-Pro-Asp-Gly-Lys-Lys-Ser-Phe-Glu-Cys- Pro-Gly-Asp-Ser-Tyr-Ile-Leu-Asp-Lys-Ala-Glu-Glu-Glu-Gly-Leu-Glu-Leu-Pro- Tyr-Ser - Cys-Arg-Ala-Gly-Ser-Cys-Ser-Ser-Cys-Ala-Gly-Lys-Val-Leu-Thr-Gly-Ser-Ile- Asp-Gln - Ser-Asp-Gln-Ala-Phe-Leu-Asp-Asp-Asp-Gln-Gly-Gly-Asp-Gly-Tyr-Cys-Leu-Thr- Cys-Val - Thr-Tyr-Pro-Thr-Ser-Asp-Val-Thr-Ile-Lys-Thr-His-Cys-Glu-Ser-Glu-Leu . It was composed of 93 amino acid residues with 7 cysteine residues, the highest number found among the chloroplast-type ferredoxins so far sequenced . A cysteine residue was found for the first time at the 89th position in a chloroplast-type ferredoxin . Calculation of the numbers of amino acid differences among chloroplast-type ferredoxins indicates that the Peridinium ferredoxin is far divergent not only from higher plant ferredoxins but also from blue-green algal ferredoxins.

J Dairy Sci, 1988 Nov, 71(11), 3143 - 7
Antibiotic treatment of mastitis: comparison of intramammary and intramammary plus intramuscular therapies; Owens WE et al.; Efficacy of two therapy regimens for subclinical Staphylococcus aureus bovine mastitis was evaluated . Forty-nine lactating cows with a total of 78 subclinically infected quarters were included . Cows were divided into two treatment groups . Group 1 received intramammary infusion at each milking for six milkings with a commercial lactating cow product containing 62.5 mg of amoxicillin . Group 2 received the same intramammary infusion regimen plus one intramuscular injection daily of 9,000,000 U procaine penicillin G for 3 d . The combination of intramammary and intramuscular treatment resulted in bacteriologic cure of 51.4% of quarters and 48% of cows compared to 25% of quarters and 30.4% of cows for intramammary infusion alone . Quarters where infections were not cured supported more growth of S . aureus during treatment than glands that were cured . Mean antibiotic concentration in fore-milk samples during treatment were lower in quarters that were cured . Somatic cell counts of milk from quarters that were not cured were higher prior to initiation of treatment than those cured . Data suggest that further evaluation of new regimens for treatment of subclinical S . aureus mastitis is warranted.

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