Staphylococcus aureus

Rinsho Byori, 1994 Dec, 42(12), 1227 - 33
{Molecular typing of the methicillin resistance determinant (mec) of clinical strains of Staphylococcus based on mec hypervariable region length polymorphisms}; Nishi J et al.; We used a method for molecular typing of the methicillin resistance determinant (mec) based on the size of the mec-associated hypervariable region amplified by the polymerase chain reaction (PCR) to examine 106 methicillin-resistant Staphylococcus aureus (MRSA), 9 methicillin-resistant (Mcr) S . epidermidis and 5 Mcr S . haemolyticus clinical isolates . In the 106 MRSA isolates, 5 sizes of PCR products were observed . The MRSA isolates were grouped into five hypervariable region (HVR) genotypes on the basis of the size of the PCR product . The PCR products amplified from 8 of 9 Mcr S . epidermidis isolates were the same as products amplified from MRSA isolates, which was confirmed by the PCR-SSCP (single-strand conformation polymorphism) method . In 32 methicillin-susceptible isolates, the target gene was not amplified . This method is thought to be useful in epidemiological investigations of nosocomial infections caused by MRSA . This is the first typing method capable of comparing the mec determinants of MRSA isolates and Mcr CNS isolates to establish the origin of the mec determinant.

Yakugaku Zasshi, 1994 Dec, 114(12), 1015 - 20
{A quantitative determination of inducibility by 14 membered ring macrolide antibiotics in inducible resistant Staphylococcus aureus to macrolide-lincosamide-streptogramin B antibiotics}; Kobayashi H et al.; Using Staphylococcus aureus ISP447 strain, which shows inducible resistance to macrolide-lincosamide-streptogramin B (MLS) antibiotics, the extent of MLS-resistance induced by several macrolide antibiotics {erythromycin (EM), oleandomycin (OL), or roxithromycin (RXM)} was determined in terms of a relative ratio of a growth rate of the induced cells in the presence of a challenging drug, rokitamycin (RKM), to that of uninduced cells in the absence of RKM . The ratio was referred to as a relative inducibility (%) . The inducibility was obtained at an optimum-induced condition by considering the following factors: (1) exponentially growing cells, (2) the optimum concentration of an inducer drug, i.e., 50, 150, and 150 ng/ml for EM, OL, and RXM, respectively, (3) a 3-h previous incubation at 37 degrees C in the presence of the inducer, and (4) 300 ng of RKM/ml, which is found to be optimum for induced cells to challenge, because of having no inducer activity . Using these qualification methods, inducibilities of EM, OL, and RXM as an inducer were 100.4, 27.9 and 81.1%, respectively . This method is allowed to be useful for the analysis of a structure-inducibility relationship.

Ophthal Plast Reconstr Surg, 1994 Dec, 10(4), 267 - 70
Hydroxyapatite orbital implant abscess: histopathologic correlation of an infected implant following evisceration; Ainbinder DJ et al.; A 60-year-old diabetic man with a history of ocular trauma and absolute glaucoma underwent evisceration with placement of an 18 mm hydroxyapatite orbital implant . The host scleral shell was left intact with no posterior opening for vascular ingrowth . One year later the patient presented with implant exposure, limited fibrovascular ingrowth into the implant, and a Staphylococcus aureus orbital abscess . The implant was removed, and pathology demonstrated suppurative inflammation with limited vascular ingrowth . This patient's risk factors for an implant-associated wound infection included diabetes, impaired wound healing, history of trauma, early implant exposure, and delayed fibrovascular ingrowth . Awareness of the infectious complications of any orbital implant including hydroxyapatite allows the surgeon to alter management strategies in an attempt to reduce such risk . Surgeons may consider posterior scleral portals with evisceration surgery to facilitate hydroxyapatite vascularization . Patient selection, implant size, and surgical technique are key factors for a management strategy designed to reduce the risk of implant infection.

Eur J Immunol, 1994 Dec, 24(12), 3237 - 40
Interleukin-8 differentially modulates interleukin-4- and interleukin-2-induced human B cell growth; Kimata H et al.; The effect of interleukin-8 (IL)-8 on human B cell growth, as determined by thymidine uptake and viable cell numbers was studied . IL-8 inhibited IL-4-induced growth of B cells costimulated with anti-mu antibodies (Ab) or Staphylococcus aureus Cowan strain I (SAC) in a dose-dependent fashion . In contrast, IL-8 did not inhibit IL-2-induced growth of B cells . The IL-8-mediated inhibition was specific, since it was blocked by anti-IL-8 mAb but not by control IgG1 . Moreover, anti-tumor necrosis factor-alpha (anti-TNF-alpha) Ab blocked IL-8-mediated inhibition . On the other hand, TNF-alpha, but not other cytokines including IL-1 beta, IL-3, IL-5, IL-6, interferon-alpha (IFN-alpha) or IFN-gamma, inhibited IL-4-mediated growth, and inhibition by TNF-alpha was blocked by anti-TNF-alpha Ab but not by control IgG . IL-4 had no effect on TNF-alpha binding by B cells while it decreased TNF-alpha production by B cells . IL-8 had no effect in binding of IL-4, IL-2 or TNF-alpha by B cells, however, it enhanced TNF-alpha production by B cells . These results indicate that IL-8 inhibited IL-4-induced human B cell growth by enhancement of endogenous TNF-alpha production.

Yan Ke Xue Bao, 1994 Dec, 10(4), 251 - 3
{The clinical observation of China-made ofloxacin eye drops in the treatment of bacterial infection of the external eye}; Lin B et al.; Ninety patients with extraocular infection were observed . After bacteriological examination on all of the patients, we found that staphylococcus epidermidis was the main pathogen and accounted for 43.3% . Next to it were staphylococcus aureus (31.1%), saprophytic staphylococcus was (5.6%), diplococcus catarrhus (3.3%) and moraxella sp . (2.2%) . All the patients were treated with Ofloxacin eye drops produced in Guangzhou . The bacteriological examination showed negative in 98.9% of the patients, which proved this medicine has high antibiotic power.

Yan Ke Xue Bao, 1994 Dec, 10(4), 241 - 3, 250
{Bacteriological detection of anterior chamber aspirate after extracapsular cataract extraction with intraocular lens implantation}; Yang W et al.; We cultured the anterior chamber aspirates of 83 patients with extracapsular cataract extraction and IOL implantation . Of the 83 cases, 5 (6.1%) had positive aspirates including 3 cases of staphylococcus epidermidis and 2 cases of staphylococcus aureus . No eye in our study developed bacterial endophthalmitis . Our study suggests that the anterior chamber of human eyes is capable of clearing a low inoculum of bacteria after cataract surgery without the development of bacterial endophthalmitis.

Minerva Chir, 1994 Dec, 49(12), 1299 - 303
{The capability of silicone prostheses to promote postoperative infections . An in-vitro study}; Arciola CR et al.; The authors have quantitatively evaluated and compared in vitro the Staphylococcus aureus and Staphylococcus epidermidis strain adhesiveness on various types of silicone used in surgery . The results show that there are some adhesiveness differences between the two assayed strains (S . epidermidis adhesiveness > S . aureus adhesiveness) and among silicone types (adhesiveness on soft silicones > adhesiveness on hard silicones) . However all silicones are suitable substrata for in vitro bacterial adhesion.

J Antimicrob Chemother, 1994 Dec, 34(6), 921 - 30
Decreased bactericidal activity during the period of the postantibiotic effect; Gudmundsson S et al.; Standard and clinical strains of Staphylococcus aureus, Escherichia coli and Klebsiella pneumoniae were subjected to continuous exposure to beta-lactams and aminoglycosides during the postantibiotic effect phase induced by rifampicin or erythromycin (for S . aureus) . A significant inhibition of bactericidal activity by these agents during the PAE period was observed . The degree of inhibition was dependent both on the class of antimicrobial agent (beta-lactams > aminoglycosides) and the microorganism (Gram-negative bacilli > S . aureus).

J Antimicrob Chemother, 1994 Dec, 34(6), 909 - 20
Role of beta-lactamase of methicillin-susceptible Staphylococcus aureus in resistance to first-generation oral cephems both in vitro and in vivo; Takenouchi T et al.; Cefaclor, among the oral cephalosporins tested, showed the largest inoculum effect with respect to MIC values for 61 clinical isolates of methicillin-susceptible Staphylococcus aureus, including 39 beta-lactamase producing strains . These 39 strains were divided into eight type A, 29 type B or C, and two type D producers, by comparisons of specific activities to three substrates . Two producers, one each of types A and C, were further studied to investigate the effect of beta-lactamase on staphylococcal resistance to several beta-lactams . Concentrations of cefaclor and cephalexin in cultures of these strains decreased rapidly, whereas hydrolysis of these drugs by the purified beta-lactamases was moderate to low as detected by spectrophotometric assay . Cefaclor showed high affinities for penicillin-binding proteins 1, 2, and 3 of both beta-lactamase producers and their respective penicillinase-non-producing mutants . In experimental intraperitoneal infections in mice, cefaclor was therapeutically effective against both mutants, showing 50% effective doses of less than 10 mg/kg/dose . In contrast, it was not satisfactory against the parent strains, requiring greater-than-10-fold increases in concentration for the same degree of survival . We concluded that resistance to first-generation oral cephems seen both in vitro and in vivo was due mainly to the beta-lactamase production.

J Antimicrob Chemother, 1994 Dec, 34(6), 899 - 907
In-vitro antibacterial activity of fusidic acid alone and in combination with other antibiotics against methicillin-sensitive and -resistant Staphylococcus aureus; Drugeon HB et al.; The activity of fusidic acid alone and in combination with gentamicin, oxacillin, rifampicin, fosfomycin and ciprofloxacin was investigated against 36 strains of Staphylococcus aureus . The fusidic acid MIC50 of 0.06 mg/L and MIC90 of 0.125 mg/L were determined for 36 strains . Bactericidal activity of fusidic acid was confirmed . Several types of interaction were observed with other antibiotics: dominant effect of fusidic acid over gentamicin; synergy with rifampicin for three or four strains; indifference or dominance with oxacillin, indifference with vancomycin, indifference or dominance with fosfomycin, and indifference with ciprofloxacin . Antagonism was seen with only one strain, following exposure to fusidic acid and rifampicin.

J Antimicrob Chemother, 1994 Dec, 34(6), 885 - 97
The effect of Triton X-100 on the in-vitro susceptibility of methicillin-resistant Staphylococcus aureus to oxacillin; Komatsuzawa H et al.; The effect of the non-ionic detergent, Triton X-100, on the in-vitro activity of oxacillin against methicillin-resistant (MRSA) and methicillin-susceptible (MSSA) strains of Staphylococcus aureus was investigated . In the presence of Triton X-100, the MICs of oxacillin for both MRSA and MSSA isolates were reduced; this enhancing effect was particularly marked for the MRSA strains . Triton X-100 therefore counteracted the resistance to methicillin encoded by mecA . In the presence of oxacillin at subinhibitory concentrations, Triton X-100 induced the bacteriolysis of MRSA and potentiated the autolysis of these organisms . However, the detergent had no effect on the bacteriolytic enzyme profile or the susceptibility of the bacterial cell wall to bacteriolytic enzymes, nor did it promote the binding of oxacillin to the penicillin-binding protein (PBP) 2A . On the other hand, it stimulated the release from the bacteria of acylated lipoteichoic acid (LTA), a putative endogenous regulator of autolysins . Autolytic enzyme-deficient MRSA mutants were equally as sensitive as the parent strain to the effect of Triton X-100 on susceptibility to oxacillin . These results indicate that the enhanced in-vitro activity of oxacillin against MRSA in the presence of Triton X-100 cannot be accounted for simply by the induction of bacteriolysis following activation of autolytic enzymes by the detergent-stimulated release of LTA.

Jpn J Pharmacol, 1994 Dec, 66(4), 451 - 6
Enhancement by ascorbic acid 2-glucoside or repeated additions of ascorbate of mitogen-induced IgM and IgG productions by human peripheral blood lymphocytes; Tanaka M et al.; In this study, the effect of ascorbic acid 2-glucoside (AA-2G), a stable derivative of ascorbic acid (AsA), or repeated additions of ascorbate on antibody productions by human peripheral blood lymphocytes (PBLs) was examined, and the physiological function of AsA was evaluated . When human PBLs were stimulated with Staphylococcus aureus Cowan I or pokeweed mitogen, AA-2G remarkably increased the numbers of IgM- and IgG-secreting cells which were detected by enzyme-linked immunospot assay . Although a single addition of ascorbate was without effect, the effect of AA-2G was remarkably inhibited by the addition of castanospermine, an alpha-glucosidase inhibitor; and moreover, repeated additions of AsA to the culture medium during the culture period enhanced the response to the same level as did a single addition of AA-2G . These results indicate that AsA has the ability to stimulate the immunoglobulin productions by AA-2G . The phytohemagglutinin-induced proliferative response of PBLs was also stimulated by AA-2G . The intracellular AsA content in PBLs cultured with AA-2G was maintained at relatively high levels during the culture period, whereas the content with a single dose of AsA reached nearly zero by the end of the experiment . These in vitro findings suggest that AA-2G and AsA function as potent immunostimulators of antibody production in humans and that the intracellular AsA content is a key parameter for establishing the immune response of PBLs.

Protein Eng, 1994 Dec, 7(12), 1463 - 70
The stability and unfolding of an IgG binding protein based upon the B domain of protein A from Staphylococcus aureus probed by tryptophan substitution and fluorescence spectroscopy; Bottomley SP et al.; The stability and unfolding of an immunoglobulin (Ig) G binding protein based upon the B domain of protein A (SpAB) from Staphylococcus aureus were studied by substituting tryptophan residues at strategic locations within each of the three alpha-helical regions (alpha 1-alpha 3) of the domain . The role of the C-terminal helix, alpha 3, was investigated by generating two protein constructs, one corresponding to the complete SpAB, the other lacking a part of alpha 3; the Trp substitutions were made in both one- and two-domain versions of each of these constructs . The fluorescence properties of each of the single-tryptophan mutants were studied in the native state and as a function of guanidine-HCl-mediated unfolding, and their IgG binding activities were determined by a competitive enzyme-linked immunosorbent assay . The free energies of folding and of binding to IgG for each mutant were compared with those for the native domains . The effect of each substitution upon the overall structure and upon the IgG binding interface was modelled by molecular graphics and energy minimization . These studies indicate that (i) alpha 3 contributes to the overall stability of the domain and to the formation of the IgG binding site in alpha 1 and alpha 2, and (ii) alpha 1 unfolds first, followed by alpha 2 and alpha 3 together.

J Virol Methods, 1994 Dec, 50(1-3), 29 - 41
Rapid coagglutination test for the detection and typing of foot and mouth disease virus; Montassier HJ et al.; Protein A containing Staphylococcus aureus was used to develop a coagglutination (COA) test for the detection and typing of foot and mouth disease virus (FMDV) O, A and C serotypes in infected cells and tissues . Different batches and amounts of guinea pig anti-FMDV sera were assessed to optimize the preparation of COA conjugates . The sensitivity and specificity of the COA Test for the detection of FMDV O, A and C serotypes and heterologous viruses was also characterized . Comparison between the COA Test and complement fixation test for the detection and typing of FMDV obtained from extracts of tongue epithelial tissues from infected cattle revealed high agreement in the results and indicated a potential application of the COA Test for the direct diagnosis of viruses.

Int J Immunopharmacol, 1994 Dec, 16(12), 977 - 84
Immunostimulating effects of protein A in immunosuppressed aflatoxin-intoxicated rats; Raisuddin S et al.; Aflatoxin B1, the potent carcinogenic compound produced by the Aspergillus flavus group of fungi on food and feed, induces immunosuppressive effects in rodents . In this communication, we report an immunomodulatory approach to abrogate aflatoxin B1-induced immunotoxicity in rats using protein A of Staphylococcus aureus Cowan 1 . We have earlier demonstrated that protein A can protect the animals from toxicities induced by a number of drugs, chemicals and toxins . In the present study various combinations of aflatoxin B1 exposure and protein A treatment in animals were used . It was observed that protein A could provide protection to animals from aflatoxin B1-induced immunotoxicity, as measured by a battery of tests assessing cell-mediated immunity (CMI) profile of the host . Various parameters showing suppression of CMI following aflatoxin B1 exposure were reverted back towards normalcy in protein A-treated animals . It is concluded that protein A may prove to be a useful agent to protect the host from aflatoxin immunotoxicity, in view of its stimulatory effects on various immune functions even after their initial depression due to aflatoxin B1.

Int J Food Microbiol, 1994 Dec, 24(1-2), 171 - 8
Enterotoxigenic Escherichia coli and Staphylococcus aureus in fish and seafood from the southern region of Brazil; Ayulo AM et al.; An investigation to evaluate the microbiological condition and safety of fish and seafood commonly harvested at the coast of Santa Catarina State and sold in Florianopolis was undertaken . One hundred and seventy-five samples of fish and fish fillets (Cynoscion leiarchus), shrimp tails (Peneaus paulensis), shellfish-meat (Anomalocardia brasiliensis and Metilus edulis), and crab-meat (Callinectes sapidus) were collected from markets and examined within 4 h of purchase . For isolation and enumeration of Escherichia coli the methods used were those of Speck et al . (1975) (Method 1) and Fishbein et al . (1976) (Method 2); for S . aureus, methods recommended by the U.S . Food and Drug Administration were used including biochemical identification of the strains . E . coli was more frequently detected with Method 1 than Method 2 . Of 317 E . coli strains tested for STG and LT II toxins, only one (isolated from shellfish-meat) produced ST and none produced LT II toxin . S . aureus was isolated from 20% of 175 samples examined, including 60% of samples of shellfish-meat . Only nine of 109 S . aureus strains produced enterotoxins, including enterotoxin A (4), D (1) and AB (4) . It is concluded that greater care must be taken to reduce contamination of fish and seafood during harvesting and post-harvest handling.

Antimicrob Agents Chemother, 1994 Dec, 38(12), 2810 - 6
Effect of magnesium complexation by fluoroquinolones on their antibacterial properties; Lecomte S et al.; By using infrared and 19F nuclear magnetic resonance spectroscopies, we localized the binding site and measured the affinity of magnesium for six fluoroquinolones . It was proven that magnesium is situated between the ketone and the carboxylate groups . We determined the binding constants for the 1:1 Mg(2+)-drug complex in solution . Sparfloxacin and pefloxacin, with affinity constants (Ka) of (10.1 +/- 0.6) x 10(2) M-1 and (21 +/- 1) x 10(2) M-1, respectively, were the least and the most bound, respectively . The trend of the affinities of the assayed fluoroquinolones for magnesium was correlated with their antimicrobial activities against four bacteria and with their accumulation by these bacteria . The reference strain, Escherichia coli KL16, and two resistant mutants, NalA (gyrase mutation) and NalB (uptake defect), plus Staphylococcus aureus 209P were used . It appeared that, in every case, an impairment of accumulation is responsible for the increase in the MICs observed upon the addition of magnesium.

Antimicrob Agents Chemother, 1994 Dec, 38(12), 2702 - 9
Pharmacodynamics of levofloxacin, ofloxacin, and ciprofloxacin, alone and in combination with rifampin, against methicillin-susceptible and -resistant Staphylococcus aureus in an in vitro infection model; Kang SL et al.; The pharmacodynamic properties of levofloxacin (an optically active isomer of ofloxacin), ofloxacin, and ciprofloxacin, alone and in combination with rifampin, were evaluated over 24 to 48 h against clinical isolates of methicillin-susceptible and -resistant Staphylococcus aureus (MSSA 1199 and MRSA 494, respectively) in an in vitro infection model . The incidence of the emergence of resistance among the test strains was also determined . The fluoroquinolones were administered to simulate dosage regimens of 200 mg, 400 mg given intravenously (i.v.) every 12 h (q12h), and 400 and 800 mg given i.v . q24h . Rifampin was dosed at 600 mg i.v . q24h . Although the MICs and MBCs of the quinolones were similar (< or = 0.49 microgram/ml), levofloxacin was the most potent agent in time-kill studies on the basis of the time required to achieve a 99.9% reduction in the number of log10 CFU per milliliter (e.g., with the regimen of levofloxacin {400 mg q24h, 6.5 h} versus ofloxacin {12.5 h}, P < 0.024, and levofloxacin versus ciprofloxacin {6.5 versus 9.0 h}, P < 0.0017) against MSSA 1199 . The killing activity of levofloxacin was similar to that of ofloxacin against MRSA 494 (time to achieve a 99.9% reduction in the number of log10 CFU per milliliter, 11.1 versus 13.8 h, respectively) . Levofloxacin and ofloxacin dosed once daily demonstrated greater bactericidal activity than when they were dosed twice daily against MSSA 1199 . Resistance to levofloxacin or ofloxacin was not observed with any dosage regimen . Furthermore, resistance to ofloxacin was not detected when the half-life was reduced from 6 to 3 h . Regrowth and stable resistance (65-fold increase in the MIC for MSSA 1199; 16-fold increase in the MIC for MRSA 494) were noted within 24 h of exposure to ciprofloxacin at 200 mg q12h . Combination therapy with rifampin prevented the emergence of resistance to ciprofloxacin . Neither DNA gyrase alteration nor an energy-dependent efflux process mediated by the norA gene appeared to be responsible for the resistance observed . Our data suggest that with levofloxacin there is a more rapid onset of bactericidal activity than with ofloxacin or ciprofloxacin against MSSA 1199 and that the activity of levofloxacin is similar to that of ofloxacin but better than that of ciprofloxacin against MRSA 494 . Resistance was noted only after exposure to the low dose of ciprofloxacin . Resistance to ofloxacin did not develop even when the pharmacokinetics of the drug were set to equal those of ciprofloxacin, suggesting that ofloxacin differs from ciprofloxacin irrespective of time of exposure . The resistance to ciprofloxacin that developed in our vitro model may be mediated by the cfx-ofx locus, which has been shown to be associated with low-level fluoroquinolone resistance . Overall, levofloxacin demonstrated potent bactericidal activity against S . aureus, without the emergence of resistance in our infection model . Quinolones dosed once daily were more effective than equivalent dosages administered twice daily . The addition of rifampin was not synergistic but prevented the emergence of ciprofloxacin resistance.

Am J Infect Control, 1994 Dec, 22(6), 340 - 5
Control of methicillin-resistant Staphylococcus aureus in a pediatric burn unit; Sheridan RL et al.; BACKGROUND: Control of methicillin-resistant Staphylococcus aureus (MRSA) is particularly difficult in burn units, which are often cited as sources of hospital-wide MRSA outbreaks . We developed a successful MRSA control program and document here its apparent effectiveness in controlling MRSA transmission in a pediatric burn unit . METHODS: An MRSA control program that included surveillance culturing, clinician feedback, flexible, site-specific isolation, and a list of known carriers was consistently applied in a pediatric burn unit through a 7-year period . Microbiology reports of MRSA isolates from patients and environmental surfaces and records of all patients from whom MRSA was isolated were reviewed . RESULTS: During calendar years 1985 through 1991, a total of 991 acutely burned children were admitted to the Boston unit of the Shriners Burns Institute . Forty MRSA cases (4%) were identified . One patient both had MRSA at admission and met our criteria for nosocomial MRSA . Of the remaining 39 patients, 11 had MRSA at admission and 28 had nosocomial MRSA . There were 17 wound infections, two cases of pneumonia, and two bloodstream infections . No deaths were attributed to MRSA sepsis . CONCLUSION: An MRSA control program including surveillance culturing, clinician feedback, flexible, site-specific isolation, and a list of known carriers is associated with a low rate of nosocomial MRSA in a pediatric burn unit.

Eur J Epidemiol, 1994 Dec, 10(6), 743 - 8
Stability of genomic DNA fragment patterns in methicillin resistant Staphylococcus aureus (MRSA) during the course of intra- and interhospital spread; Witte W et al.; The analysis of genomic DNA fragment patterns has revealed as a powerful tool for strain discrimination in Staphylococcus aureus; for use as an epidemiological marker, stability during the course of an outbreak is an essential prerequisite . Genomic DNA fragment patterns (SmaI restriction, pulsed-field electrophoresis) of four different epidemic MRSA strains were compared along with intra- and interhospital and country-wide spread over more than 12 months in Germany . Strain I was isolated from infections in 8 hospitals . In one hospital a subclone arised which differed from the original strain by 4 fragments . Strain II was spread among 4 hospitals, isolates from three of these hospitals exhibited a variability of one to three fragments in the 150-200 kb range . Two hospitals in the Hannover-area were affected by strain III; in 17 isolates of this strain a variability up to three fragments was found in the 170-200 kb range . Strain IV was isolated from 19 cases of infections in 3 hospitals in Berlin . The fragment patterns were completely stable . When S . aureus strains are typed by genomic DNA fragment patterns, a variability in a definite range of molecular masses during the course of an epidemic should be taken into consideration.

Rev Saude Publica, 1994 Dec, 28(6), 406 - 9
Staphylococcal food poisoning from cream-filled cake in a metropolitan area of south-eastern Brazil; Pereira ML et al.; Twelve people became ill with vomiting and diarrhoea approximately four hours after eating cake with a cream filling at a birthday party and on the day following . The cake had been prepared by a food handler who had long experience in preparing foods for such functions . Staphylococcus aureus that produced enterotoxin A was isolated from the nose, the fingernails, and a healed infection on the neck of the food handler, and from the cake . Enterotoxin A was detected in the remaining portion of the cake . The cake, while still warm, had been refrigerated for one hour after it was prepared before it was removed for the party; it was refrigerated after the party . The cake was large (6 kg) and hence it was not adequately cooled in the hour during which it was refrigerated before the party . The conclusion is that the cake was accidentally contaminated by the food handler and inadequately cooled before it was eaten.

Mol Pharmacol, 1994 Dec, 46(6), 1048 - 55
Agonist-induced photoincorporation of a p-benzoylphenylalanine derivative of substance P into membrane-spanning region 2 of the Torpedo nicotinic acetylcholine receptor delta subunit; Blanton MP et al.; The neuropeptide substance P acts, at micromolar concentrations, as a noncompetitive antagonist of nicotinic acetylcholine receptors (AChRs) of both neuronal and muscle subtypes . The mechanism of this inhibition has been shown to be most consistent with stabilization of a nonconducting desensitized state of the AChR, via binding to a site distinct from both the agonist site and the high affinity noncompetitive antagonist site . We have used a radioiodinated photoreactive analogue of substance P, containing the amino acid p-benzoyl-L-phenylalanine in place of the Phe8 residue of substance P, to identify the sites of interaction of substance P within the Torpedo california AChR . AChR-rich membrane suspensions were photolabeled in the absence or presence of the agonist carbamylcholine and/or nonradioactive substance P, and incorporation into AChR subunits was assessed by autoradiography after sodium dodecyl sulfate-polyacrylamide gel electrophoresis . In the absence of agonist 125I incorporation was detected in each subunit and was insensitive to substance P, whereas in the presence of carbamylcholine there was a 2-fold increase in photoincorporation into the AChR delta subunit that was inhibited by the addition of an excess of substance P . The sites of specific photoincorporation in the delta subunit were initially mapped by use of Staphylococcus aureus V8 protease to a 14-kDa fragment extending from delta Ile-192 to Glu-280 . Further fragmentation of this 14-kDa fragment with trypsin and S . aureus V8 protease established that the sites of specific incorporation were restricted to the region delta Ser-253 to Glu-280, which contains the membrane-spanning region 2 that is known to form the lining of the ion channel . These results establish that in the presence of agonist at least a part of the undecapeptide substance P binds within the ion channel in the desensitized state of the AChR, and it is likely that the binding of substance P to this site is responsible for the action of substance P as a noncompetitive AChR antagonist.

Leukemia, 1994 Dec, 8(12), 2083 - 94
Expression and regulation of CD30 ligand and CD30 in human leukemia-lymphoma cell lines; Gruss HJ et al.; The CD30 antigen was originally described as a specific surface marker for Hodgkin's lymphoma . Recent work established CD30 as a member of the tumor necrosis factor/nerve growth factor receptor superfamily whose ligand (CD30L) has also been cloned and expressed; CD30L is active as membrane-bound type II glycoprotein . Here, CD30L mRNA expression was studied in a panel of 102 continuous human leukemia-lymphoma cell lines and was found only in four Burkitt lymphoma, one Burkit-type acute lymphoblastic leukemia and one non-Hodgkin's lymphoma (NHL) cell line . The product of CD30L mRNA is expressed as a membrane protein on the surface of these malignant B-cell lines . Treatment of these cell lines with soluble CD27L, phorbol ester or staphylococcus aureus Cowan antigen resulted in the enhancement of cell surface CD30L protein expression . CD30L mRNA was not detected in normal unstimulated peripheral blood (PB) monocytes, monocyte-derived macrophages, or T-cells, but was detected in primary granulocytes; exposure to activating reagents induced and upregulated CD30L transcription in these different PB populations . While CD40 and CD30L surface protein expression on PB monocytes could be enhanced or induced by treatment with gamma-interferon, these cells remained negative for CD30, both at the mRNA and at the protein level . Similarly, PB monocyte-derived macrophages and granulocytes remained negative for CD30 mRNA and protein expression, regardless of stimulation . Only activated T-cells expressed CD30 mRNA and surface protein . CD30L-transfected cells and cells constitutively expressing CD30L delivered a similar stimulus for proliferation of the CD30+ Hodgkin's disease (HD)-derived cell line HDLM-2, but inhibited proliferation of the CD30+ large cell anaplastic lymphoma cell line KARPAS-299 . These data provide strong evidence for the involvement in growth regulation of recombinant and natural CD30L through its interaction with the CD30 receptor . Collectively, these data suggest that the CD30L-CD30 interaction has potent biological activity and might play a critical role in the immune response and pathogenesis of HD and some NHL, in particular Burkitt lymphomas.

Biochemistry, 1994 Nov 22, 33(46), 13938 - 45
Identification of the neuronal acceptor in bovine cortex for ammodytoxin C, a presynaptically neurotoxic phospholipase A2; Krizaj I et al.; A specific, high-affinity binding site for ammodytoxin C in synaptic membranes from bovine cerebral cortex was detected and partially characterized . Equilibrium binding analysis revealed a single population of {125I}ammodytoxin C acceptors with the following binding parameters: Kd = 6.0 nM and Bmax = 5.7 pmol/mg membrane protein . Such binding was strongly inhibited by three ammodytoxins (A, B, and C) and by crotoxin B . Vipera berus berus phospholipase A2 was a weaker inhibitor; nontoxic phospholipase A2, ammodytin I2, and the myotoxic phospholipase A2 homologue, ammodytin L, both from Vipera ammodytes ammodytes venom, inhibited binding only at very high concentrations, whereas alpha-dendrotoxin, beta-bungarotoxin, and crotoxin A had no influence on the {125I}ammodytoxin C-specific binding . The ammodytoxin C neuronal binding site therefore overlaps, at least partially, with the neuronal acceptors for some of the related presynaptically neurotoxic phospholipases A2 (beta-neurotoxins) . {125I}-Ammodytoxin C was covalently attached to its acceptor by chemical cross-linking . Subsequent SDS-PAGE analysis followed by autoradiography revealed saturably labeled membrane components with apparent M(r) values of 51,000 (weaker band) and 53,000-56,000 (stronger band) . Pretreatment of synaptic membranes with Staphylococcus aureus V-8 proteinase and proteinase K, heat, or low pH decreased the {125I}ammodytoxin C-specific binding to various extents, but never abolished it completely . Membrane protein and certain phospholipids residing in its vicinity are therefore most likely involved in the binding of ammodytoxin C to bovine synaptic membranes.

J Biol Chem, 1994 Nov 18, 269(46), 29121 - 8
Conformational differences in human apolipoprotein B-100 among subspecies of low density lipoproteins (LDL) . Association of altered proteolytic accessibility with decreased receptor binding of LDL subspecies from hypertriglyceridemic subjects; Chen GC et al.; We asked at what point in the metabolic cascade of very low density lipoproteins (VLDL) to low density lipoproteins (LDL) the accessibility of proteolytic cleavage sites in B-100 changes, and we evaluated the effect of hypertriglyceridemia on the proteolytic accessibility, secondary structure, and receptor-binding affinity of B-100 in LDL subspecies of varying density . Limited proteolysis with Staphylococcus aureus V8 protease and cathepsin D identified the density (about 1.033 g/ml) between two LDL subspecies, designated LDL-1 and -2, as the transition point during VLDL metabolism of both normolipidemic (N-) and hypertriglyceridemic (HTG-) subjects at which accessibility to protease attack changed in three peptide regions of B-100 . Hypertriglyceridemia greatly altered proteolytic accessibility of B-100 in the denser LDL subspecies . Specifically, B-100 in HTG-LDL exposed more cleavage sites than in N-LDL, including two novel sites, approximately 120 and approximately 130 kDa from the NH2 terminus in the small and dense subspecies (designated LDL-4, -4.5 or -5, d = 1.048-1.062 g/ml) . Analysis of circular dichroic spectra indicated no difference in helical content between B-100 in N- and HTG-LDL but showed a greater content of beta-structure in HTG-LDL . Binding affinity for the LDL receptor of human fibroblasts decreased markedly with increasing density among HTG-LDL subspecies (by approximately 50% for LDL-4.5 or -5) . We conclude that the changes in proteolytic accessibility observed between LDL-1 and -2 and in LDL-4, -4.5, or -5 indicate significant differences in local conformation of B-100 at specific peptide regions . The association of exposure of more cleavage sites, especially novel sites in the NH2-terminal regions, with greatly decreased receptor-binding affinity in LDL-4.5 or -5 suggests that altered local conformation in B-100 apart from the putative receptor-binding domain might affect interaction with the receptor.

J Biol Chem, 1994 Nov 4, 269(44), 27246 - 50
Reduced methicillin resistance in a new Staphylococcus aureus transposon mutant that incorporates muramyl dipeptides into the cell wall peptidoglycan; Ornelas-Soares A et al.; Screening of a new Tn551 library constructed in the background of a highly methicillin-resistant Staphylococcus aureus strain identified a new insertion site located on the SmaI B-fragment of the chromosome that reduced the minimal inhibitory concentration of the parent (1600 micrograms/ml) to 25-50 micrograms/ml in the mutant and caused heterogeneous expression of resistance and abnormality in peptidoglycan composition (absence of the unsubstituted pentapeptide and incorporation of alanylglutamate- and alanylisoglutamate-containing muropeptides) . There was an accumulation of large amounts of the UDP-linked muramyl dipeptide in the cytoplasmic wall precursor pool of the mutant . Reduced (heterogeneous) antibiotic resistance and all the biochemical abnormalities were reproduced in genetic backcrosses by transduction with phage 80 alpha . Mutant RUSA235 appears to be impaired in the biosynthesis of the staphylococcal cell wall precursor muropeptide before the lysine addition step . We propose to provisionally call the gene inactivated in this mutant femF.

Bull Tokyo Dent Coll, 1994 Nov, 35(4), 217 - 20
Detection of methicillin-resistant staphylococcus aureus in human saliva and on denture surfaces; Honma K et al.; The prevalence of Staphylococcus species and methicillin-resistant Staphylococcus aureus (MRSA) in human saliva and on denture surfaces was examined with selective media . A total of 166 saliva samples from dental students and individuals using dentures and 39 swab samples from denture surfaces were subjected in this study . The detected percentage of MRSA from saliva samples was 2.3% . Five out of 39 swab samples of denture surface contained MRSA or methicillin-resistant Staphylococcus epidermidis (MRSE) . The isolated MRSA and MRSE were resistant to beta-lactam antibiotics.

J Appl Bacteriol, 1994 Nov, 77(5), 549 - 52
Staphylococcal growth and enterotoxins (A-D) and thermonuclease synthesis in the presence of dehydrated garlic; Gonzalez-Fandos E et al.; The inhibition of Staphylococcus aureus growth and enterotoxin and thermonuclease production by various concentrations of garlic (Allium sativum) was studied in BHI broth . The growth of Staph . aureus was inhibited by dehydrated garlic at levels of 1.5% (w/v) and over . Enterotoxins A, B and C1 were only detectable in broth containing < 1% of garlic while enterotoxin D was produced at a level of 2% . Garlic also inhibited thermonuclease (TNAse) production, complete inhibition being observed at levels > or = 1.5% . TNAse was not always detected when enterotoxin was present.

J Antibiot (Tokyo), 1994 Nov, 47(11), 1266 - 72
Aldecalmycin, a new antimicrobial antibiotic from Streptomyces . I . Taxonomy, fermentation, isolation, physico-chemical and biological properties; Sawa R et al.; A new antibiotic, aldecalmycin, has been discovered in the culture broth of Streptomyces sp . MJ147-72F6 . Aldecalmycin was purified by solvent extraction, Diaion HP-20 chromatography, silica gel chromatography, Sephadex LH-20 chromatography, HPLC and centrifugal partition chromatography . The 1H and 13C NMR spectra of aldecalmycin showed the presence of keto-enol tautomers . Aldecalmycin is equipotent in inhibiting the growth of sensitive and methicillin-resistant Staphylococcus aureus (MRSA).

J Antibiot (Tokyo), 1994 Nov, 47(11), 1250 - 7
Pyrroindomycins, novel antibiotics produced by Streptomyces rugosporus sp . LL-42D005 . I . Isolation and structure determination; Ding W et al.; Pyrroindomycins A and B were isolated from fermentations of culture LL-42D005, a strain of Streptomyces rugosporus . Pyrroindomycins possess potent antimicrobial activities against methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococci . Their structures have been determined by using 1- and 2-D NMR, mass spectroscopy and chemical degradations . Pyrroindomycins are the first natural products that contain the highly unsaturated pyrroloindole moiety.

Br J Dermatol, 1994 Nov, 131(5), 717 - 9
Juvenile pityriasis rubra pilaris associated with hypogammaglobulinaemia and furunculosis; Castanet J et al.; We report a case of follicular keratosis with inflammatory changes, consistent with a diagnosis of atypical juvenile pityriasis rubra pilaris . An unusual feature was the occurrence of severe Staphylococcus aureus folliculitis and furunculosis, a phenomenon rarely encountered in pityriasis rubra pilaris and the other follicular keratoses . Standard antibiotic and antiseptic treatment for chronic S . aureus infection was ineffective . The patient was subsequently found to have hypogammaglobulinaemia, and treatment with human polyvalent immunoglobulin infusions was successful in eradicating the sepsis . It is therefore probable that the hypogammaglobulinaemia played a pathogenic role in the development of cutaneous sepsis.

Medicine (Baltimore), 1994 Nov, 73(6), 299 - 305
Pacemaker endocarditis . Report of 44 cases and review of the literature; Arber N et al.; We conducted a retrospective study to characterize the clinical course, microbiologic spectrum, and risk factors for endocarditis and for associated mortality in a large series of patients with documented pacemaker endocarditis . Using a computerized search through the medical records of 10 major hospitals in Israel from 1982 to 1992, and carefully reviewing the charts, we identified 44 patients with pacemaker endocarditis . The cases were categorized as definite (n = 25), probable (n = 12), or possible (n = 7) infective endocarditis based on strict case definition . Fever and chills were the most common symptoms . Increased ESR, leukocytosis, microscopic hematuria, and anemia were the most common laboratory findings . A relatively high proportion of the patients were diabetic . The most common source of endocarditis was infection acquired by the placement procedure or infection of the pacemaker pouch . Demographic, clinical, and laboratory features were similar to those of endocarditis patients of a similar age range without pacemakers, although the frequency of fever and chills was higher in our patients than in those patients and splenomegaly, vascular embolic phenomena, and new or changing murmurs were rare in our patients . The major pathogens were Staphylococcus aureus and Staphylococcus epidermidis, similar to other series of pacemaker-associated bacteremia and similar to the microbiologic findings of early prosthetic-valve endocarditis . However, this microbiologic profile is different from that of native-valve endocarditis . Although the present series did not show a statistically significant advantage to electrode removal over conservative treatment, when analyzed together with pooled data from other studies, it suggests that the surgical approach is preferable.(ABSTRACT TRUNCATED AT 250 WORDS)

J Bone Joint Surg Br, 1994 Nov, 76(6), 955 - 9
Influence of type of medullary nail on the development of local infection . An experimental study of solid and slotted nails in rabbits; Melcher GA et al.; Any operation involving the implantation of a foreign body increases the risk of infection . The implant material and its surface, the dead space, and any necrosis or vascular changes play a significant role in susceptibility to infection . We investigated the effect of the dead space in an intramedullary nail on the rate of local infection . We inoculated the intramedullary cavities of rabbit tibiae with various concentrations of a human pathogen, of Staphylococcus aureus strain, and then inserted either a solid or a hollow slotted stainless-steel nail . We found a significantly higher rate of infection after use of the slotted nail (59%) than after the solid nail (27%) (p < 0.05).

Am J Physiol, 1994 Nov, 267(5 Pt 1), L578 - 84
Binding of surfactant protein A to C1q receptors mediates phagocytosis of Staphylococcus aureus by monocytes; Geertsma MF et al.; During both steady-state conditions and inflammatory reactions in the lower airways, monocytes migrate to the alveoli where they come into contact with surfactant . Surfactant is composed of phospholipids, neutral lipids, and specific proteins, and its main function is to reduce surface tension in the alveoli . The most abundant glycoprotein surfactant protein A (SP-A) affects the structure, function, and metabolism of pulmonary surfactant . The aim of the present study was to determine whether SP-A plays a role in the antibacterial activities of human monocytes and whether this is mediated by a receptor for SP-A on these cells . The results showed that SP-A binds to both Staphylococcus aureus and monocytes and mediates the phagocytosis of the bacteria by these cells . SP-A does not stimulate the intracellular killing of bacteria by monocytes, and SP-A-opsonized S . aureus do not induce the production of reactive oxygen intermediates . SP-A binds to the C1q receptor (C1qR) on monocytes, since its binding was inhibited by C1q and the SP-A-enhanced association of S . aureus with these cells was completely abolished when monocytes were adherent to surfaces coated with C1q or anti-C1qR monoclonal antibody . Furthermore, the binding of SP-A to monocytes results in an increased intracellular concentration of adenosine 3',5'-cyclic monophosphate . Together, these results demonstrate that C1qR mediates the phagocytosis of SP-A-opsonized S . aureus by monocytes.

Surg Neurol, 1994 Nov, 42(5), 424 - 7
Spinal subdural abscess: successful treatment with limited drainage and antibiotics in a patient with AIDS; Sathi S et al.; The authors present a case of Staphylococcus aureus spinal subdural abscess in a patient with AIDS . Although complete surgical drainage has been strongly advocated in the literature, this patient made a complete neurologic and symptomatic recovery and radiographically demonstrated resolution of the abscess with only limited surgical drainage and parenteral antibiotics . Magnetic resonance imaging findings of this unusual lesion are discussed . Relevant literature in the management of spinal subdural abscesses is reviewed.

Surg Neurol, 1994 Nov, 42(5), 417 - 23
HIV status does not affect microbiologic spectrum or neurologic outcome in spinal infections; Heary RF et al.; The impact of human immunodeficiency virus (HIV) on the clinical presentations, causative organisms, and neurologic outcomes of patients with spinal infections is reviewed . Thirty-two patients with spinal epidural abscesses, vertebral osteomyelitis, or both were treated at an urban hospital over a 42-month period . Thirteen of these patients were confirmed by serologic analysis to be HIV seropositive . The diagnoses were confirmed by 30 open surgical procedures (14 anterior, 16 posterior) and seven percutaneous biopsies . Twenty-seven intraoperative cultures were positive and the remaining three patients had positive blood cultures prior to the surgical procedure . In both the HIV (+) and HIV (-) groups, Staphylococcus aureus predominated as the causative organism (overall rate: 72%) . Mycobacterium tuberculosis was the second most common organism . The clinical presentations in both groups were similar with pain as the most frequent symptom and objective neurologic abnormalities on physical examination in 29 of the 32 patients (91%) . The results of this analysis show that the clinical presentations and organisms cultured do not differ depending upon a concurrent HIV infection . Furthermore, the ultimate neurologic outcome of patients with spinal infections depends on their neurologic status at the time of treatment and not on their HIV status.

J Gen Virol, 1994 Nov, 75 ( Pt 11), 2897 - 909
Structural protein relationships among eastern equine encephalitis viruses; Strizki JM et al.; We have re-evaluated the relationships among the polypeptides of eastern equine encephalitis (EEE) viruses using SDS-PAGE and peptide mapping of individual virion proteins . Four to five distinct polypeptide bands were detected upon SDS-PAGE analysis of viruses: the E1, E2 and C proteins normally associated with alphavirus virions, as well as an additional more rapidly-migrating E2-associated protein and a high M(r) (HMW) protein . In contrast with previous findings by others, the electrophoretic profiles of the virion proteins of EEE viruses displayed a marked correlation with serotype . The protein profiles of the 33 North American (NA)-serotype viruses examined were remarkably homogeneous, with variation detected only in the E1 protein of two isolates . In contrast, considerable heterogeneity was observed in the migration profiles of both the E1 and E2 glycoproteins of the 13 South American (SA)-type viruses examined . Peptide mapping of individual virion proteins using limited proteolysis with Staphylococcus aureus V8 protease confirmed that, in addition to the homogeneity evident among NA-type viruses and relative heterogeneity among SA-type viruses, the E1 and E2 proteins of NA- and SA-serotype viruses exhibited serotype-specific structural variation . The C protein was highly conserved among isolates of both virus serotypes . Endoglycosidase analyses of intact virions did not reveal substantial glycosylation differences between the glycoproteins of NA- and SA-serotype viruses . Both the HMW protein and the E2 protein (doublet) of EEE virus appeared to contain, at least in part, high-mannose type N-linked oligosaccharides . No evidence of O-linked glycans was found on either the E1 or the E2 glycoprotein . Despite the observed structural differences between proteins of NA- and SA-type viruses, Western blot analyses utilizing polyclonal antibodies indicated that immunoreactive epitopes appeared to be conserved.

J Exp Med, 1994 Nov 1, 180(5), 1675 - 83
Characterization and biological properties of a new staphylococcal exotoxin; Ren K et al.; Staphylococcus aureus strain D4508 is a toxic shock syndrome toxin 1-negative clinical isolate from a nonmenstrual case of toxic shock syndrome (TSS) . In the present study, we have purified and characterized a new exotoxin from the extracellular products of this strain . This toxin was found to have a molecular mass of 25.14 kD by mass spectrometry and an isoelectric point of 5.65 by isoelectric focusing . We have also cloned and sequenced its corresponding genomic determinant . The DNA sequence encoding the mature protein was found to be 654 base pairs and is predicted to encode a polypeptide of 218 amino acids . The deduced protein contains an NH2-terminal sequence identical to that of the native protein . The calculated molecular weight (25.21 kD) of the recombinant mature protein is also consistent with that of the native molecules . When injected intravenously into rabbits, both the native and recombinant toxins induce an acute TSS-like illness characterized by high fever, hypotension, diarrhea, shock, and in some cases death, with classical histological findings of TSS . Furthermore, the activity of the toxin is specifically enhanced by low quantities of endotoxins . The toxicity can be blocked by rabbit immunoglobulin G antibody specific for the toxin . Western blotting and DNA sequencing data confirm that the protein is a unique staphylococcal exotoxin, yet shares significant sequence homology with known staphylococcal enterotoxins, especially the SEA, SED, and SEE toxins . We conclude therefore that this 25-kD protein belongs to the staphylococcal enterotoxin gene family that is capable of inducing a TSS-like illness in rabbits.

J Leukoc Biol, 1994 Nov, 56(5), 661 - 5
Isolation of antimicrobial peptides from avian heterophils; Evans EW et al.; Five bactericidal peptides (chicken heterophil peptides CHP1 and CHP2; turkey heterophil peptides THP1, THP2, and THP3) were purified from avian heterophil granules . All peptides were cationic and rich in cysteine, arginine, and lysine . The complete amino acid sequence, consisting of 39 amino acids, was determined for CHP1 . This peptide had a molecular weight of 4481 as determined by mass spectrometry . Partial NH2-terminal amino acid sequences were obtained for the remaining peptides . Both chicken peptides and THP1 shared sequence homology at 22 residues and a cysteine motif which was similar to that of bovine beta-defensins . THP2 and THP3 were homologous to each other but were not homologous to the other three and had a unique cysteine motif . Peptides CHP1, CHP2, and THP1 killed Staphylococcus aureus and Escherichia coli in vitro, whereas THP2 and THP3 killed only S . aureus in vitro.

J Clin Invest, 1994 Nov, 94(5), 1815 - 22
Diminished virulence of a sar-/agr- mutant of Staphylococcus aureus in the rabbit model of endocarditis; Cheung AL et al.; Microbial pathogenicity in Staphylococcus aureus is a complex process involving a number of virulence genes that are regulated by global regulatory systems including sar and agr . To evaluate the roles of these two loci in virulence, we constructed sar-/agr- mutants of strains RN6390 and RN450 and compared their phenotypic profiles to the corresponding single sar- and agr- mutants and parents . The secretion of all hemolysins was absent in the sar-/agr- mutants while residual beta-hemolysin activity remained in single agr- mutants . The fibronectin binding capacity was significantly diminished in both single sar- mutants and double mutants when compared with parents while the reduction in fibrinogen binding capacity in the double mutants was modest . In the rabbit endocarditis model, there was a significant decrease in both infectivity rates and intravegetation bacterial densities with the double mutant as compared to the parent (RN6390) at 10(3)-10(6) CFU inocula despite comparable levels of early bacteremia among various challenge groups . Notably, fewer bacteria in the double mutant group adhered to valvular vegetations at 30 min after challenge (10(6) CFU) than the parent group . These studies suggest that both the sar and agr loci are involved in initial valvular adherence, intravegetation persistence and multiplication of S . aureus in endocarditis.

Exp Parasitol, 1994 Nov, 79(3), 468 - 79
Leishmania major: association of the differentially expressed gene B protein and the surface lipophosphoglycan as revealed by membrane capping; Pimenta PF et al.; The lipophosphoglycan (LPG) of Leishmania promastigotes forms a dense glycocalyx which effectively covers the entire surface of the cell, and which undergoes structural modifications during the differentiation of promastigotes to the infective or metacyclic stage . Recently, the first protein marker for metacyclic promastigotes of Leishmania major has been characterized . This protein, termed gene B protein, is located on the cell surface, yet it lacks any hydrophobic sequence for membrane attachment . It does contain an unusual amino acid repeat that is related to the peptidoglycan binding domain of protein A from Staphylococcus aureus, suggesting that the protein might interact with metacyclic LPG via this domain for attachment to the cell . We have studied the distribution of LPG, gene B protein, and the major surface protease, gp63, by labeling them with immunogold or immunofluorescence prior to and during capping events . Thin sections of double-labeled parasites revealed that the gene B protein-gold particles were colocalized with the LPG-gold particles in the LPG capping structures at the extremities of the cell . Cocapping of LPG and gene B protein was also observed with two-color fluorescence . No similar redistribution was seen in gp63 or with integral membrane proteins . In contrast to the gene B protein, gp63 could only be immunogold labeled on the metacyclic surface after capping and shedding of the LPG, providing further that it and other membrane-associated proteins are normally buried under the LPG coat . The unusual surface exposure of the gene B protein is consistent with its hydrophilic and LPG binding properties, which allow it to become incorporated into the cell coat and to localize to the most external aspects of the cell.

EMBO J, 1994 Nov 1, 13(21), 5245 - 51
Replication-specific conversion of the Staphylococcus aureus pT181 initiator protein from an active homodimer to an inactive heterodimer; Rasooly A et al.; The Staphylococcus aureus rolling circle plasmid pT181 regulates its replication by controlling the synthesis of its initiator protein RepC . RepC is inactivated during pT181 replication by the addition of an oligodeoxynucleotide, giving rise to a new form, RepC* . We analyzed RepC and RepC* in four classes of mutants: plasmid copy number mutants, two classes of RepC mutants affecting different portions of the protein and oriC (origin) mutants . We have found that in the cell with wild-type RepC there are similar relative amounts of RepC and RepC*, regardless of copy number, and that the conversion of RepC to RepC* is replication dependent . Genetic and biochemical evidence is presented that RepC functions as a dimer and that during replication the RepC homodimer is converted to the RepC/RepC* heterodimer.

Biochemistry, 1994 Nov 1, 33(43), 12844 - 51
Mutations affecting the activity of toxic shock syndrome toxin-1; Deresiewicz RL et al.; Toxic shock syndrome toxin-1 (TSST-1), the potent staphylococcal exoprotein linked to most cases of the toxic shock syndrome, is a V beta-restricted T-cell mitogen (a so-called "superantigen") . TSST-ovine (TSST-O) is a natural variant of TSST-1, and is produced by certain ovine mastitis-associated strains of Staphylococcus aureus . Compared to TSST-1, TSST-O is only weakly mitogenic for leporine or murine splenocytes . It differs from TSST-1 at 7 amino acid residues over its 194 amino acid length . Terminus shuffling between the two proteins has suggested that their C-terminal differences (T69, Y80, E132, and I140 in TSST-1; 169, W80, K132, and T140 in TSST-O) are in part responsible for their discrepant mitogenic properties . In order to explore further the functional consequences of altering TSST-1 at residues 132 and 140, we engineered point mutants of TSST-1 at those positions . The mutant proteins were purified to homogeneity from culture supernants of a nontoxigenic strain of S . aureus using a combination of ultrafiltration, liquid-phase isoelectric focusing, and ion-exchange chromatography . The mutants retained global structural integrity as evidenced by circular dichroism spectroscopy, their preserved resistance to trypsin digestion, and their preserved binding to a neutralizing murine monoclonal antibody . The mutants were then tested for mitogenicity for human T-cells: The mutant I140T was approximately as active as wild-type TSST-1, while the mutant E132D was about 10-fold attenuated . On the other hand, the mutants E132A or E132K were each at least 1000-fold attenuated.(ABSTRACT TRUNCATED AT 250 WORDS)

J Am Acad Dermatol, 1994 Nov, 31(5 Pt 1), 746 - 54
Cutaneous findings in HIV-1-positive patients: a 42-month prospective study . Military Medical Consortium for the Advancement of Retroviral Research (MMCARR); Smith KJ et al.; BACKGROUND: Cutaneous disease is common in patients infected with HIV-1 . OBJECTIVE: The aim of our study was to identify cutaneous markers associated with HIV-1 infection and disease progression as measured by Walter Reed (WR) stage . METHODS: For 42 months we have observed 912 HIV-1-positive patients in all WR stages . All patients had an extensive past and present medical history taken as well as a complete physical examination, periodic visits, and appropriate diagnostic procedures . RESULTS: Increasing dryness of the skin and seborrheic dermatitis are early findings in a large percentage of patients in WR stage 1; the occurrence and severity of both conditions increase with disease progression . Tinea infections, condylomata acuminata, and verrucae are seen early, but with disease progression, although there is no clear increase in occurrence, these infections become more diffuse and resistant to treatment . Flares in acne vulgaris and folliculitis show a peak occurrence in early and mid-stage disease with a decreased occurrence in late-stage disease . Herpes simplex infections, oral candidiasis, molluscum contagiosum, Staphylococcus aureus infections, and oral hairy leukoplakia show a marked increase in occurrence with advanced disease . Conditions that have a statistically significant association with disease progression as measured by a change in a stage include drug eruptions, seborrheic dermatitis, oral candidiasis, oral hairy leukoplakia, molluscum contagiosum, herpes zoster, and hyperpigmentation (nail, oral, skin) . CONCLUSION: The most frequent and persistent cutaneous disorders were asteatosis (with or without asteatotic eczema) and seborrheic dermatitis . Conditions that were associated with a change in WR stage include drug eruptions, seborrheic dermatitis, oral candidiasis, oral hairy leukoplakia, molluscum contagiosum, herpes zoster, and hyperpigmentation . In addition to Kaposi's sarcoma, patients with HIV-1 disease have an increased potential for the development of both cutaneous epithelial and probably melanocytic malignancies . Epithelial tumors were seen in patients in all stages of disease.

Plasmid, 1994 Nov, 32(3), 295 - 305
A conjugative transfer system for the rumen bacterium, Butyrivibrio fibrisolvens, based on Tn916-mediated transfer of the Staphylococcus aureus plasmid pUB110; Clark RG et al.; A limitation of genetic studies of the rumen bacterium, Butyrivibrio fibrisolvens, has been the availability of suitable vectors and transfer systems . Using the conjugative tetracycline resistant transposon, Tn916, the Staphylococcus aureus plasmid, pUB110, and the pUB110-based shuttle vector, pUBLRS, a conjugative transfer system was developed for B . fibrisolvens . B fibrisolvens donor strains H17c2 and H17c12, containing Tn916 and pUB110 or pUBLRS, respectively, were used in mating experiments with selected B . fibrisolvens strains . Kanamycin resistant transconjugants, containing pUB110, of strains 193, 194, and 195 were detected at a combined average frequency of 7.78 x 10(-7) per donor and 1.11 x 10(-5) per recipient . Transconjugants of strains 193 and 194, containing pUBLRS, were detected at an average frequency of 1.22 x 10(-6) per donor and 4.70 x 10(-8) per recipient . Southern hybridization analysis confirmed the presence of pUB110 and pUBLRS in transconjugants . Results indicated that Tn916 was necessary for mobilization of pUB110 as transconjugants were not detected when the transposon was absent from the donor strains . The ability to mobilize pUB110 and pUBLRS between B . fibrisolvens strains provides a conjugative transfer system that circumvents problems encountered with electroporation.

J Pharm Pharmacol, 1994 Nov, 46(11), 892 - 5
Flavanones with potent antibacterial activity against methicillin-resistant Staphylococcus aureus; Iinuma M et al.; With the therapeutic concept of using the defensive ability of plants against microbial infections, phytoalexin, an antimicrobial phytochemical was studied for its ability to inhibit the growth of methicillin-resistant Staphylococcus aureus (MRSA) . Extracts from Sophora exigua (Leguminosae) were fractionated by serial chromatography and the anti-MRSA activity of each fraction was determined by the agar-plate method . Among the active isolates, 5,7,2',6'-tetrahydroxy-6-isoprenyl-8-lavandulyl-4'-methox yflavanone (exiguaflavanone D) completely inhibited the growth of all the MRSA strains examined at the concentration of 1.56-6.25 micrograms mL-1, and 5, 2',6'-trihydroxy-8-lavandulyl-7-methoxy-flavanone (exiguaflavanone B) inhibited at a concentration of 50 micrograms mL-1 . This former compound is expected to be a phytotherapeutic agent for MRSA infections as an alternative to conventional antibiotics with unwanted side-effects or the appearance of antibiotic-resistant bacteria.

Clin Infect Dis, 1994 Nov, 19(5), 970 - 2
Infectious complications associated with temporary epidural catheters; Pegues DA et al.; To identify cases of deep-tissue or local infection associated with temporary epidural catheters, we reviewed medical records from 1980 through 1992 and prospectively followed up patients with temporary epidural catheters from January 1993 through June 1993 who were hospitalized at a large, tertiary referral hospital . We identified seven cases of temporary epidural catheter-associated infection, including three cases of deep-tissue infection (paraspinal muscle abscess, epidural abscess, and meningitis) and four cases of local infection . The infections were diagnosed 2 days to 22 days following insertion of the epidural catheter . Staphylococcus aureus was isolated in four (57%) of the cases . All seven patients were treated with removal of the catheter and antimicrobial therapy; three patients also required surgical drainage for management of deep-tissue infection . Our findings emphasize the importance of daily inspection and prompt removal of temporary epidural catheters if infection is suspected.

Diagn Microbiol Infect Dis, 1994 Nov, 20(3), 171 - 4
Comparison of mupirocin susceptibility of nasal and nonnasal Staphylococcus aureus isolates; Utrup LJ et al.; Susceptibilities of 414 nasal and 586 nonnasal Staphylococcus aureus isolates, both methicillin resistant and methicillin susceptible, to the topical antimicrobial agent mupirocin were compared . A susceptibility of 99.1% was observed for the 1000 isolates . Nasal and nonnasal isolates showed a similar 90% minimum inhibitory concentration (MIC90) and statistically equivalent percent susceptibilities.

FEMS Immunol Med Microbiol, 1994 Nov, 10(1), 47 - 53
Vaccination with Staphylococcus aureus fibrinogen binding proteins (FgBPs) reduces colonisation of S . aureus in a mouse mastitis model; Mamo W et al.; A mouse mastitis model was used to study the effect of vaccination with fibrinogen binding proteins and collagen binding protein from Staphylococcus aureus against challenge infection with S . aureus . The mice vaccinated with fibrinogen binding proteins showed reduced rates of mastitis compared with controls . Gross examination of challenged mammary glands of mice showed that the glands of mice immunized with fibrinogen binding proteins developed mild intramammary infection or had no pathological changes compared with glands from control mice . Histopathological examination of tissue sections from challenged glands showed that most glands from mice vaccinated with fibrinogen binding protein developed disseminated necrosis or had no pathological changes . A significantly reduced number of bacteria could be recovered in the glands from mice immunized with fibrinogen binding proteins as compared with controls . In a similar study, immunization of mice with collagen binding protein did not induce protection against challenge infection with S . aureus.

Antimicrob Agents Chemother, 1994 Nov, 38(11), 2590 - 8
Reassessment of the number of auxiliary genes essential for expression of high-level methicillin resistance in Staphylococcus aureus; de Lencastre H et al.; A new transposon library constructed in the background of the highly and homogeneously methicillin-resistant Staphylococcus aureus strain COL yielded 70 independent insertional mutants with reduced levels of antibiotic resistance . Restriction analysis with HindIII, EcoRV, EcoRI, and PstI and then Southern hybridization with probes for the transposon and for the femA-femB gene demonstrated that 41 of the 70 Tn551 mutants carried distinct and novel, as yet undescribed insertion sites, all of which were outside of the mecA gene and were also outside the already-characterized auxiliary genes femA, femB, femC, and femD . All previously described Tn551 mutations of this type were in genes located either on SmaI fragment A or SmaI fragment I . In contrast, inserts of the new library were located in 7 of the 16 SmaI chromosomal fragments, fragments A, B, C, D, E, F, and I . In all of the mutants, expression of methicillin resistance became heterogeneous, and the MIC for the majority of cells was reduced (1.5 to 200 micrograms ml-1) from the homogeneous methicillin MIC (1,600 micrograms ml-1) of the parental cells . Although identification of the exact number of genes inactivated through the new set of transposon inserts will require cloning and sequencing, a rough estimate of this number from mapping data suggests a minimum of at least 10 to 12 new genetic determinants, all of which are needed together with femA, femB, femC, and femD for the optimal expression of methicillin resistance.

Antimicrob Agents Chemother, 1994 Nov, 38(11), 2568 - 71
Altered production of penicillin-binding protein 2a can affect phenotypic expression of methicillin resistance in Staphylococcus aureus; Hackbarth CJ et al.; Altered production of penicillin-binding protein 2a (PBP 2a) may affect the phenotypic expression of resistance in methicillin-resistant Staphylococcus aureus (MRSA) . COL, an MRSA strain that constitutively produces PBP 2a, was transformed with a recombinant plasmid containing the two beta-lactamase regulatory genes, blaI and blaR1, with either the beta-lactamase gene, blaZ, or a truncated blaZ . Both of the transformed MRSA strains now produced an inducible PBP 2a, and the MICs of nafcillin, methicillin, and imipenem for these strains were similar to those for the parental strain . A mutation in blaR1 that resulted in the complete repression of PBP 2a production altered the phenotypic expression of methicillin resistance in that strain, as evidenced by efficiency-of-plating experiments . Rather than being homogeneously resistant like COL, the blaR1 mutant strain now appeared to have a small resistant subpopulation . Gene products that regulate PBP 2a production may contribute to the organism's expression of methicillin resistance, but additional chromosomally located factors are required.

Anticancer Res, 1994 Nov-Dec, 14(6B), 2633 - 6
Effects of amino and imino acridines on tumor necrosis factor production by human leukocytes; Mandi Y et al.; Tumor necrosis factor (TNF) is a multifunctional cytokine with diverse effects on different cells and tissues . The biological activity of TNF is described on the basis of its cytotoxic action in vivo and in vitro . Different acridines were systematically synthesized and their effects were tested on endotoxin and Staphylococcus aureus-induced TNF production by human leukocytes . 9-aminobutylacridine and 9-ethylaminoacridine totally abrogated the TNF production of leucocytes at a concentration of 3.5 microM, whereas 9-imino -10-butylacridine and 9-imino-10-ethylacridine exerted only a 50% inhibition in the same concentration . Derivatives designated as 9-amino-(2-dimethylamino-ethyl)-acridine and 9-imino-10-(2-dimethylamino-ethyl)-acridine in a concentration of 7 microM exerted only a 30% and a 10% inhibition respectively . A significant modulation of TNF production was not observed when other alkylated derivatives in this series were applied . The TNF-mediated cytotoxic effect of monocytes against WEHI cells was also reduced by the most effective compounds . The acridines did not interfere with the expression of CD 14 molecules on monocytes . The exact mechanism of the suppression of TNF synthesis by acridines remains to be elucidated, but might be useful in the screening and evaluation of their anticancer properties and antimalarial effects.

Mol Cell Endocrinol, 1994 Nov, 105(2), 217 - 26
Arginine vasopressin (AVP) causes the reversible phosphorylation of the myristoylated alanine-rich C kinase substrate (MARCKS) protein in the ovine anterior pituitary: evidence that MARCKS phosphorylation is associated with adrenocorticotropin (ACTH) secretion; Liu JP et al.; We have recently shown that AVP causes a protein kinase C (PKC)-dependent increase in ACTH release and biosynthesis in ovine anterior pituitary cells . In these cells, AVP also causes the translocation of PKC from the cytosol to the cell membrane which is maximal at 5 min, but the intracellular events distal to protein kinase C activation that underlie ACTH secretion have not been well characterized to date . Since the MARCKS protein has been implicated in neurosecretion and is phosphorylated by PKC in synaptosomes, studies were carried out to determine whether AVP might cause MARCKS phosphorylation in the ovine anterior pituitary, and to determine whether this phenomenon might be temporally correlated with PKC translocation and the release of ACTH . When cytosolic fractions of rat brain, ovine anterior pituitary, and cultured ovine anterior pituitary cells were incubated with purified PKC, several proteins were phosphorylated including those in the region of 83-85 kDa . After precipitation of the proteins with 40% acetic acid, the 83-85 kDa phosphoproteins were selectively recovered in the acid soluble phase . Phosphopeptide maps of either the 83 or 85 kDa proteins were generated with Staphylococcus aureus V8 protease and revealed 13 and 9 kDa phosphopeptides, which are characteristic of the authentic MARCKS protein . An identical phosphopeptide map was also obtained when the MARCKS protein was selectively extracted from intact 32P-labeled anterior pituitary cells . MARCKS phosphorylation was markedly increased when ovine anterior pituitary cells were exposed to 1 microM phorbol 12-myristate 13-acetate (PMA) . When the cells were exposed to 1 microM AVP, MARCKS phosphorylation increased at 15 s and reached the maximal plateau value at 30 s . MARCKS phosphorylation then started to diminish at 2 min, and baseline levels were attained by 10 min . In the same cells, AVP stimulated ACTH release in a biphasic manner-during the first 30 s, there resulted a rapid burst of ACTH secretion that was followed by a slower, but sustained rate of secretion . We conclude that: (1) AVP causes a rapid, and reversible, phosphorylation of the MARCKS protein in the ovine anterior pituitary; (2) since the AVP-induced increase in MARCKS phosphorylation occurs much earlier in these cells than does PKC trans-location, MARCKS phosphorylation may provide a more sensitive index of the onset of PKC activation than the translocation assay; (3) the close temporal association between MARCKS phosphorylation and the rapid early release of ACTH suggests that MARCKS phosphorylation may be involved in the initial intracellular events that underly exocytosis of the hormone.

East Afr Med J, 1994 Nov, 71(11), 736 - 8
Absence of neutropenia in African patients with AIDS and associated pyomyositis; Ansaloni L et al.; The association between AIDS and pyomyositis was recently pointed out in temperate and tropical countries . In Western countries, the patients affected by pyomyositis associated with AIDS in most cases are neutropenic . We compare a group of 17 patients with pyomyositis and AIDS living in temperate climates from the literature, and 11 patients affected by the same association seen by us in northern Uganda . The patients from Western countries were significantly more neutropenic and their mean of the neutrophil count was significantly lower when compared with our group . We suggest that the defective neutrophil function associated with HIV infection play a major role in the pathogenesis of pyomyositis in our patientsPIP: Pyomyositis is an infection of the skeletal muscle mostly caused by Staphylococcus aureus . Patients in Western countries affected by pyomyositis associated with AIDS tend to be neutropenic . The objective of this study was to find out whether neutropenia is a common feature of African AIDS patients with pyomyositis; therefore, an African sample was compared with a sample of Western patients obtained from the international literature . During January-December 1993, 30 patients with pyomyositis were admitted to the Surgical Ward of Dr . Ambrosoli Memorial Hospital in Kalongo, Uganda . 11 (36.6%) were found positive for HIV and were in stage IV of the disease . The mean age was 28 years, 3 were females and 8 were males . The neutrophil count was performed before the surgical evacuation of muscle abscess . In the international literature, eight reports of 17 patients with AIDS and pyomyositis were located during the period of 1988-92 . All were males in stage IV with a mean age of 37.3 years; 8 were treated with zidovudine . When the neutrophil count was below 3000/cu . mm, the patient was considered neutropenic . 12 out of 17 Western patients were neutropenic, as opposed to only 1 out of 11 Ugandan patients (p 0.01) . The mean neutrophil count of Western patients was 3547; that of Ugandan patients was 9077 (p 0.01) . Neutrophils are primary effector cells in host defense against staphylococcal infections such as pyomyositis; hence, it has been suggested that neutropenia contributes to the development of pyomyositis in AIDS patients . Neutrophils from AIDS patients were demonstrated to be defective in their ability to kill Staphylococcus aureus in vitro, compared with neutrophils from seronegative controls . Therefore, the defective neutrophil function associated with HIV infection contributed to the development of pyomyositis in Ugandan patients, among whom neutropenia was not common .

Infect Control Hosp Epidemiol, 1994 Nov, 15(11), 703 - 9
Colonization and transmission of high-level gentamicin-resistant enterococci in a long-term care facility; Chenoweth CE et al.; OBJECTIVES: To assess the prevalence of high-level gentamicin-resistant enterococcus (HGRE) colonization, transmission patterns, and spectrum of illness among residents of a long-term care facility . DESIGN: Monthly surveillance for HGRE colonization of wounds, rectum, and perineum over a 1-year period . SETTING: A Veterans Affairs long-term care facility attached to an acute-care facility . PATIENTS: All 341 patients in the facility during the observation period . RESULTS: Over the 1-year period, 120 patients (35.2%) were colonized with HGRE at least once, with an overall monthly colonization rate of 20 +/- 1.5% . HGRE were isolated from rectum (12.8%), wounds (11.7%), and perineum (9.3%) . Patients with the poorest functional status had the highest rate of colonization (P < 0.0005) . HGRE-colonized patients were more likely to be colonized with methicillin-resistant Staphylococcus aureus (51% versus 25%; P < 0.0005) . Seventy-four patients (21.7%) were colonized at admission or at the start of the study . Another 46 patients (13.5%) acquired HGRE during the study, including 36 who acquired HGRE while in the long-term care facility and 10 who were positive when transferred back from the acute-care hospital . Based on plasmid profiles, only two patients appeared to have isolates similar to those of current or previous roommates . Carriage of HGRE was transient in most cases . Only 20 patients were colonized for 4 or more months, and those patients usually carried different strains intermittently . Infections were infrequent, occurring in only 4.1% of total patients . CONCLUSIONS: In our long-term care facility, HGRE were endemic, and new acquisition of HGRE occurred frequently . However, only two patients had evidence of acquisition from a roommate, suggesting that cross-infection from a roommate was not a major route of spread of HGRE.

Rinsho Byori, 1994 Nov, 42(11), 1182 - 7
{Detection of methicillin-resistant Staphylococcus aureus using PCR and non-radioactive DNA probes: IV . Mutational sequences in the region upstream of the mec A gene in clinical staphylococcal strains}; Yamashita K et al.; Expression of mec A is regulated by two regulatory genes (mecR1 and mecI); the intact regulatory genes exist in the N315 strain of methicillin-sensitive S . aureus, whereas the repressor gene (mecI) is deleted in the MR108 strain of methicillin-resistant S . aureus (MRSA) . However, our previous findings that clinical strains of N315 type were methicillin-resistant suggest possible mutations within the operator region, by which the inhibitory effect of MecI is released to induce the constitutive expression of mec A . Therefore, we analyzed nucleotide sequences in the region upstream of the mec A gene and in the regulatory genes, by using PCR-direct sequencing and ASO probes . Analysis of selected clinical strains by ASO probes revealed that 74% of MRSA and 36% of MRSE (methicillin-resistant S . epidermidis) lacked the mecI gene whereas 26% of MRSA and 64% of MRSE possessed the intact regulatory genes . Furthermore, PCR-direct sequencing identified four different mutations in the operator nucleotide sequence and one mutation in Shine-Dalgarno sequence . Additionally, these mutations were shown to occur in the strains of N315 type with higher probability rather than in those of MR108 type . The data suggest that one or more of these mutational sequences at least may have some influence on the expression of methicillin-resistance in clinical staphylococcal strains.

Kansenshogaku Zasshi, 1994 Nov, 68(11), 1421 - 7
{A case of TSS complicated with SSSS in an adult with liver cirrhosis}; Toyota E et al.; This paper reports a case of TSS complicated with SSSS in an adult with liver cirrhosis . A 52-year-old male, heavy drinker, was referred to our clinic complaining lumbago and painful swelling of the right arm . The patient had peeling of the skin over the hips, knees and elbows with positive Nikolsky's sign . The patient was in a state of shock on admission . Pyrexia persisted for 4 days and finally the body temperature rose up to 39 degrees C . The laboratory studies revealed hypoxia, DIC and multiple organ failure, and these became progressively worse . He died 4 days after admission . According to the criteria, he was diagnosed as TSS, and TSST-1 was detected from his serum . Staphylococcus aureus, coagulase type V was cultured both from the blood and from the wound of his right middle finger . This isolated strain did not produce TSST-1 . The skin specimen at autopsy showed that the cleavage plane lied at the subcorneal region and close to the granular layer, with specific changes caused by exfoliative toxin . It was compatible to the exfoliation which was caused by exfoliative toxin produced from the S . aureus coagulase type V . The autopsy also revealed alcohol liver injury, liver cirrhosis and multiple organ failure due to shock state . SSSS is rare in adults, to our knowledge this is the first reported case of TTS complicated with SSSS.

Nippon Ika Daigaku Zasshi, 1994 Nov, 61(6), 563 - 71
{Studies on the relationship between gastric acidity and the development of MRSA . Especially for the prevention of MRSA enterocolitis}; Suzuki S; Enterocolitis caused by methicillin-resistant Staphylococcus aureus (MRSA) has recently been recognized as one of the severe postoperative complications in surgery on the digestive organs . This disease often occurs in the early days after gastrointestinal operation, especially after gastrectomy . MRSA enterocolitis seems to occur when MRSA has first infected the naso-pharyngeal mucosa preoperatively, and then moved into the stomach, and subsequently proliferated in the higher pH gastric juices . The aim of this experiment was to reveal the relationship between the acidity of gastric juices and bacterial growth in the stomach during the pre and post operative period in an effort to prevent of MRSA enterocolitis . In vitro, MRSA was cultured for various periods at various pH values, and its proliferation was observed . MRSA did not grow in the culture at pH 1 at all, neither did it grow at pH 2 when cultured for more than 8 hours . This data shows the germicidal effect of high acidity in the stomach . Clinically, twenty patients with cancer in the digestive tract had the bacteria in their gastric juices examined in terms of acidity before and after operation . In cases with an increased pH level in the gastric juices after the operation . S . aureus including MRSA, was isolated frequently from the stomach . In vitro, incubation of MRSA with gastric juices collected from those cases showed no development of MRSA when the pH was below 3.98 . In order to prevent the onset of MRSA enterocolitis, the remnant stomachs of ten patients with stomach cancer were filled with hydrochloric acid lemonade just after operative reconstruction.(ABSTRACT TRUNCATED AT 250 WORDS)

Chirurg, 1994 Nov, 65(11), 1023 - 7
{Arthroscopic distension irrigation in acute postoperative infection of the knee joint--long-term follow-up}; Riel KA et al.; Between 1983 and 1993, 26 patients, 12 patients from our clinic and 14 referred patients, had to be treated because of a septic knee joint following open ligament repair or subsequent arthroscopic surgery . Immediately after arthrocentesis for synovial-fluid analysis a distension-irrigation system was established under arthroscopic control as described by R.W . Jackson . Three tubes were placed into the knee joint cavity . One was used for inflow and the other two for outflow . Distension was performed by open inflow and closed outflow tubes each second hour for 10 to 15 min . Systemic antibiotics were administered . In 22 aspirated materials the cultures revealed bacteria: mostly staphylococcus aureus (65%) . In two cases in which irrigation of the knee-joint infection was performed later than the 10th postoperative day a relapse of the infection occurred and required a complete debridement and synovectomy . During follow-up period three patients underwent second look arthroscopy due to meniscus tear, rupture of an ACL reconstruction and synovial proliferation . 7 +/- 3 (2-13) years after treatment, 24 patients were re-examined, all had x-rays and 11 patients had a MRI of the knee joint additionally . 19 patients showed excellent or good results: no complaints, full ROM, no signs of osteoarthritis on x-rays and nearly normal participation in sports 4 out of 11 patients had signs of cartilage lesion and synovial proliferation . This retrospective study indicates that the distension-irrigation system is successful only in early acute cases of knee-joint infections of postoperative origins.

Br Heart J, 1994 Nov, 72(5), 495 - 7
Persistent root abscess after emergency repair with an aortic homograft; Ritter M et al.; A fifty eight year old man with Marfan's syndrome and an aortic composite graft with a Bjork-Shiley mechanical prosthesis presented with a large aortic root abscess caused by Staphylococcus aureus endocarditis . Despite extensive surgical debridement and implantation of an aortic homograft as a composite graft, early postoperative transoesophageal echocardiography continued to demonstrate a large aortic root abscess and the patient died in a septic shock.

J Dairy Sci, 1994 Nov, 77(11), 3354 - 64
Ecology of Staphylococcus aureus isolated from various sites on dairy farms; Roberson JR et al.; The purposes of this study were to identify sources of Staphylococcus aureus on dairies and to determine whether S . aureus colonization of heifer body sites increases the risk of S . aureus IMI at parturition . In herds with high (> 10%) or low (< 3%) prevalence of S . aureus IMI, S . aureus was isolated from heifer teat skin, heifer external orifices, housing, feedstuffs, humans, nonbovine animals, air, and equipment . Additionally, in herds with high prevalence, S . aureus was isolated from bedding, insects, and water . The predominant sources of S . aureus for both groups were other IMI and heifer body sites . Heifers with prepartum lacteal secretions with S . aureus were at greater risk of S . aureus IMI at parturition than were prepartum heifers with lacteal secretions that were negative for S . aureus . Heifers with teat skin colonized by S . aureus were 3.34 times more likely to have S . aureus IMI at parturition than were noncolonized heifers . Overall, 35% of 700 heifers were colonized with S . aureus on a body site at least once . Although colonizations of most body sites appeared to be transient, a few heifers were colonized on the same site for 1 yr . Persistently colonized heifers may represent the primary reservoirs of S . aureus for other heifers.

J Dairy Sci, 1994 Nov, 77(11), 3347 - 53
Efficacy of intramuscular oxytetracycline as a dry cow treatment for Staphylococcus aureus mastitis; Erskine RJ et al.; To determine the efficacy of intramuscular oxytetracycline as a supplemental dry cow treatment for Staphylococcus aureus mastitis, 37 Holstein cows were randomly assigned to two treatment groups: intracisternal infusion with a commercial preparation of cephapirin benzathine at drying off (20 cows) and infusion with cephapirin benzathine at drying off and intramuscular oxytetracycline at 11 mg/kg once daily on d 7, 8, 9, and 10 after drying off (17 cows) . Milk samples collected 7, 14, 30, and 60 d after calving were plated for bacterial isolation within 24 h after collection and after 24 to 72 h of storage at -20 degrees C . Quarters were defined as infected if S . aureus was isolated from the fresh and frozen cultures from any one sample collected before drying off . An infected quarter was defined as cured if S . aureus was not isolated from the fresh or frozen culture from milk samples obtained following calving . The rate of cure by 30 d after calving for systemic oxytetracycline (in combination with cephapirin treatment) was 29.4% for infected quarters and 29.4% for infected cows, compared with 27.5 and 25.0%, respectively, for the cephapirin treatment only . Results including the culture at 60 d after calving were 21.2 and 22.5%, respectively, for combination therapy and cephapirin therapy only . Systemic oxytetracycline, in combination with intramammary dry cow treatment, did not improve the rate of cure for S . aureus mastitis.

Microbiology, 1994 Nov, 140 ( Pt 11), 3139 - 44
Evidence for feedback (trans) regulation of, and two systems for, glycine betaine transport by Staphylococcus aureus; Stimeling KW et al.; Previous reports are in conflict as to the number of transport systems for glycine betaine in Staphylococcus aureus . Cells grown in complex medium exhibited a single transport system of moderate affinity . Cells grown in defined medium in the absence of glycine betaine showed a high affinity and a low affinity transport system . Cells grown in the presence of glycine betaine in the presence of osmotic stress in either complex or defined media accumulated large pools of internal glycine betaine . Smaller, but still significant, amounts of glycine betaine were accumulated by cells grown in its presence in either complex or defined media in the absence of osmotic stress . Cells grown in defined medium in the presence of glycine betaine in the presence or absence of osmotic stress showed lower rates of glycine betaine transport than cells grown in its absence . This suggests that glycine betaine transport is subject to feedback or trans inhibition by internal glycine betaine . This can explain the difference in observed kinetics in cells grown in complex or defined media, the high affinity system being predominantly inhibited in cells grown in complex medium.

Microbiology, 1994 Nov, 140 ( Pt 11), 3131 - 8
Glycine betaine transport by Staphylococcus aureus: evidence for feedback regulation of the activity of the two transport systems; Pourkomailian B et al.; The regulation of glycine betaine accumulation by Staphylococcus aureus was investigated . The accumulation of glycine betaine was regulated by the osmotic pressure of the medium and the low affinity transport system played the major role in this regulation . Mutants were isolated that lack the low affinity, osmotically activated glycine betaine/proline transport system . Such mutants accumulated glycine betaine via the high affinity system but the glycine betaine pool was smaller and responded poorly to osmotic pressure changes . The regulation of glycine betaine transport has revealed that at the steady state net influx is reduced and that this is achieved by inhibition of both the low affinity and the high affinity transport systems . Cells pre-loaded with glycine betaine exhibited a reduced Vmax for both systems: the low affinity system was reduced in activity fivefold and the high affinity system was reduced 10-fold and became virtually undetectable . Although glycine betaine transport at the steady state is reduced, retention of the compatible solute is an active process since addition of an uncoupler provokes rapid release of the accumulated material . These data suggest that feedback regulation of the activity of the uptake systems is a major mechanism for controlling the level of compatible solute accumulation.

Rev Med Suisse Romande, 1994 Nov, 114(11), 1035 - 43
{Computer alert and quality of care: application to the surveillance of hospital infections}; Safran E et al.; The Centre Informatique of Geneva University Hospital is developing, in the environment of its hospital information system, DIOGENE, a computerized alert system for surveillance of hospital infections . This hospital information system is based on an open distributed architecture and a relational database system, and covers many medical applications . This environment allows the development of alerts useful for detecting patients at risk . The alerts offer to clinicians a mean to control their efficacy in patient care . They are a new application of telematics for surveillance in clinical epidemiology, and are a tool for quality assurance . Two examples of alerts established for hospital infection control activities are presented . The first alert systematically detects all cases of patients colonized by or infected with methicillin-resistant Staphylococcus aureus (MRSA) . The second alert helps to organize prospective surveillance of bloodstream infections in order to identify some risk factors for infection and propose preventive measures.

Photochem Photobiol, 1994 Nov, 60(5), 421 - 6
Spectroscopy and photosensitization of sapphyrins in solutions and biological membranes; Roitman L et al.; A spectroscopic and photophysical study of three new sapphyrin molecules is presented . The sapphyrin backbone that was derivatized to make them water soluble possesses an absorption band around 700 nm, a desired property for biological photosensitization . We studied the absorption and fluorescence spectra, from which evidence for aggregation in solvents of different polarities was obtained . The extent of aggregation is correlated with the nature of the attached moiety . The absolute quantum yields of singlet oxygen production were measured, with 1,3-diphenyl isobenzofuran as a model target, and were 0.13-0.18 in ethanol . The binding constants to liposomes and to cells were determined spectroscopically and were found to correspond to the hydrophobicities of the compounds, with an additional effect, ascribed to the sugar moiety, which was found in the case of one of the sapphyrins . The efficiency of photodamage to Staphylococcus aureus by sapphyrins and hematoporphyrin was equivalent, on the basis of cells killed per microgram of sensitizer in the incubation mixture.

Mol Gen Mikrobiol Virusol, 1994 Nov-Dec, (6), 3 - 8
{Cloning and expression of the phosphatidylinositol-specific phospholipase C gene from Listeria monocytogenes}; Ermolaeva SA et al.; The gene for phosphatidylinositol-specific phospholipase C (PI-PLC) of Listeria monocytogenes has been cloned and shown to be expressed in Escherichia coli cells from own as well as from the lactose gene promoter . The recombinant plasmid has been constructed on the basis of pRIT2T vector and carries the hybrid gone . 3-end of which is a fragment of protein A gene of Staphylococcus aureus . 3-end is a gene for phospholipase plcA, both in the same reading frame . The resultant construction is shown to code in Escherichia coli cells for the hybrid recombinant protein A:Pl-PLC . Purified preparation of the hybrid protein and polyclonal rabbit antiserum to it were obtained . The obtained antiserum to the hybrid protein containing phospholipase as en C-end domain has been shown to react specifically to phospholipase in Escherichia coli recombinant strain harbouring the constructed recombinant plasmid as well as the one in the culture fluid of listeria.

J Struct Biol, 1994 Nov-Dec, 113(3), 191 - 8
Imaging the spatial distribution of membrane receptors during neutrophil phagocytosis; Kindzelskii AL et al.; Optical microscopy and image processing have been employed to study the distribution of several cell surface receptors on living human neutrophils during opsonin-dependent and opsonin-independent phagocytosis . Receptors were labeled using fluorescein-, rhodamine-, or AMCA-conjugated F(ab')2 fragments of anti-Fc gamma RIIIB (CD16), anti-CR3 (CD11b/CD18), and anti-uPAR (urokinase-type plasminogen activator receptor) antibodies, intact phycoerythrin-labeled interleukin 8, and fluorescein- or rhodamine-labeled Con A (concanavalin A), Boc-PLPLP (tert-butyl-oxycarbonyl-Phe(D)-Leu-Phe(D)-Leu-Phe-OH), and N-formyl-Nle-Leu-Phe-Nle-Tyr-Lys . Labeled neutrophils were observed during the phagocytosis of IgG-opsonized erythrocytes and nonopsonized latex beads, Escherichia coli, and Staphylococcus aureus . To quantitate receptor distribution, cells were divided into four quadrants with the first being the point of attachment and the fourth being opposite the point of attachment . Ligated formyl peptide receptors, and to a lesser extent CR3, accumulated at the sites of target internalization for all forms of phagocytosis examined . However, Fc gamma RIIIB, uPAR, IL-8, Con A, and the FPR antagonist FBoc-PLPLP were not polarized on cells during phagocytosis . These data suggest that agonist-labeled formyl peptide receptors may play a broader role in leukocyte function than previously suggested, including possible participation in phagocytosis.

Antibiot Khimioter, 1994 Nov, 39(11), 29 - 36
{Additional differentiation of methicillin resistant strains of Staphylococcus aureus typed by the international phage bank}; Zyeva VS et al.; In the methicillin resistant strains of Staphylococcus aureus (MRSA) typed by the International Set phages the host specificity of the restriction-modification of the phage 85 DNA was determined, the finger printing of the cell DNA was carried out with using two probes and the lytic spectrum of the phages induced in them was studied . Four clones with different specificity of the restriction-modification system (rm89, rm108, rm121 and rm947) differing from that of strain PS 85 which is the host of phage 85 were detected . The strains belonging to the modification types m89, m108 and m121 contained prophages (within the respective groups) with similar lytic spectra when tested with the use of the PS strains of the International Collection and had cross antiphage immunity . Six phage variants were detected among the phages induced in the strains with the modification type m947 which could be indicative of the clone heterogeneity.

Zentralbl Bakteriol, 1994 Nov, 281(4), 495 - 501
Role of antibodies against fibronectin-, collagen-binding proteins and alphatoxin in experimental Staphylococcus aureus peritonitis and septicaemia in neutropenic mice; Rozalska B et al.; We have investigated the protective role of hyperimmune rabbit IgG against two surface structures of Staphylococcus aureus, i.e . fibronectin-, and collagen-binding proteins as well as alpha-toxin in experimental peritonitis and septicaemia in neutropenic mice pretreated with cyclophosphamide . This treatment markedly decreased clearance of bacteria from mouse organs . With combined immunotherapy given passively bacteria were eradicated more efficiently for all animals sampled, comparative to controls.

Intensive Care Med, 1994 Nov, 20 Suppl 4, S17 - 22
Anti-infective treatment in intensive care: the role of glycopeptides; Gruneberg RN et al.; Antibiotics are used in 80% of patients in the ICU, encouraging nosocomial infections with resistant organisms . If the antibiotic susceptibilities of the pathogen are known, a narrow-spectrum antibiotic is preferable to preserve the patient's resistance to colonization . However, treatment is often empirical and broad-spectrum combinations are commonly used . Gram-positive bacteraemia is associated with invasive monitoring or intravascular catheters . If the device cannot be removed easily, the glycopeptides are the only agents likely to be active against most strains of the commonest pathogen, the coagulase-negative staphylococcus . Long-stay patients are susceptible to infection with enterococci and methicillin-resistant Staphylococcus aureus, which are often resistant to all the usual agents other than glycopeptides . Vancomycin is long established, but is nephrotoxic, requires serum monitoring, must be administered as an infusion and can cause red man syndrome . Teicoplanin can be given as a single daily bolus without similar side-effects or monitoring . In deep-seated staphylococcal infection, the usual dose of teicoplanin is adequate if given in combination with other agents, but it may need to be doubled if used as monotherapy . Monitoring of the levels in the serum is helpful to ensure an adequate dose in patients with renal failure or in drug abusers, but is not needed to prevent toxicity.

Infection, 1994 Nov-Dec, 22(6), 395 - 400
Infections related to the menstrual cycle . A study of five otherwise healthy women with recurrent abscesses and a review of the literature; Weischer M et al.; The purpose of the study was to investigate the in vitro bactericidal function of blood polymorphonuclear leucocytes (PMN) in various phases of the menstrual cycle from otherwise healthy women with recurrent cutaneous abscesses related to the premenstrual phase of the menstrual cycle compared with the bactericidal activity of PMN from healthy women with no inconveniences related to the menstrual cycle . The bactericidal activity against Staphylococcus aureus 502A was investigated and when possible against the patients' own strain . No variation in bactericidal activity was observed during the different phases of the menstrual cycle . PMN from five women with recurrent abscesses related to the premenstrual phase tended to kill fewer S . aureus 502A than PMN from three women in the control group . The literature of immunological defence mechanisms and the occurrence of infections related to the menstrual cycle is reviewed.

Infection, 1994 Nov-Dec, 22(6), 390 - 4
Modern topical glucocorticoids and anti-infectives for superinfected atopic eczema: do prednicarbate and didecyldimethylammoniumchloride form a rational combination?
Korting HC, Zienicke H, Braun-Falco O, Bork K, Milbradt R, Nolting S, Schopf E, Tronnier H.
The addition of an anti-infective to a topical glucocorticoid preparation for superinfected atopic eczema is still controversial . To address this question in the context of the topical glucocorticoids of the non-halogenated double-ester type 0.25% prednicarbate cream was compared to the identical preparation incorporating the same amount of the disinfectant didecyldimethylammoniumchloride in patients suffering from atopic eczema carrying Staphylococcus aureus at a density of more than 10(6) colony-forming units per cm2 . One of the preparations was used twice daily over 5 days according to a random plan in a blind fashion . Thereafter treatment was based on either prednicarbate cream or the corresponding vehicle according to clinical needs . Clinical and microbiological evaluation were scheduled for days 0, 6 and 34 . Various clinical parameters were addressed individually as well as over all improvement using scores . A total of 143 patients was recruited . The patients of both groups improved rapidly with respect to clinical and microbiological findings . Essentially, there was no difference between the groups . Hence, the addition of an anti-infective to a topical prednicarbate preparation is not to be generally recommended.

Stem Cells, 1994 Nov, 12(6), 626 - 37
Production of functional myeloid cells from CD34-selected hematopoietic progenitor cells using a clinically relevant ex vivo expansion system; Lill MC et al.; There is increasing clinical interest focused on ex vivo manipulation and expansion of hematopoietic cells . In this study, we demonstrate that a simple combination of growth factors can expand progenitors to yield functional myeloid cells . Furthermore, this system can produce mature, functionally competent cells in the absence of fetal bovine serum (FBS), which will enhance the clinical utility of this approach . Hematopoietic progenitor cells obtained from normal bone marrow and from leukapheresis products were studied . The mononuclear fraction was enriched for CD34 cells using the Ceprate CD34 biotin kit (CellPro #LC34-1 or LC34-2) . The selected cells were expanded for two weeks in Iscove's medium supplemented with 20% FBS and various combinations of interleukin-3 (IL-3), granulocyte colony-stimulating factor (G-CSF), stem cell factor (SCF) and interleukin -6 (IL-6) added either simultaneously or sequentially . The optimal combination of these factors identified for myeloid expansion was simultaneous addition of IL-3, SCF and G-CSF (at 50 ng/ml each), resulting in an average 773 +/- 133-fold expansion of nucleated cells (n = 5) . When corrected for the purity of CD34 cells in the starting population, the mean fold expansion with IL-3, SCF and G-CSF was 2,265 +/- 729 . A mean of 74.7 +/- 10.5% (n = 3) of the expanded cells was positive for CD11b; 86-91% (n = 2) of the cells were promyelocytes or more mature granulocytes . Functional assays demonstrated normal phagocytosis and intracellular killing of Staphylococcus aureus (S . aureus) by the expanded cell population . Studies performed using cells expanded in defined serum-free media demonstrated that fold expansion was decreased and that the cells produced were less mature and functionally less competent than cells expanded with FBS . The decreased expansion could be partially reversed, and the functionality almost completely restored by the addition of autologous plasma.

Aten Primaria, 1994 Oct 31, 14(7), 892 - 4
{Prevalence of asymptomatic carriers of pathogenic germs in the nasopharynx . Comparison made between primary health care professionals and the population seeking care}; Chicote Feo R et al.; OBJECTIVE . To determine if the prevalence of non-symptomatic nasopharyngeal carriers of pathogenic germs is higher in health workers of primary care than in the general population . DESIGN . Cross-sectional, observational study . SETTING . Primary Care . Salamanca Urban Area . PARTICIPANTS . Seventy-six Health Centre workers from Salamanca and 152 individuals representing the general population attending the Health Centers were studied . People with pathology related to the study subject or with inmunitary problems were excluded . MEASUREMENTS AND MAIN RESULTS . A nasopharyngeal sample was carried out using a swab . The relative prevalence of carriers (sanitary workers vs general population) was 0.38 (c.i . 0.20-0.72) for Staphylococcus aureus and 0.56 (c.i . 0.34-0.93) for all pathogens . No significant differences were found according to sex . CONCLUSIONS . The prevalence of non-symptomatic nasopharyngeal carriers is higher in general population than in the group of primary care workers.

Arch Intern Med, 1994 Oct 24, 154(20), 2330 - 5
Staphylococcus aureus endocarditis at a community teaching hospital, 1980 to 1991 . An analysis of 106 cases; Watanakunakorn C; BACKGROUND: The clinical diagnosis of infective endocarditis due to Staphylococcus aureus can be difficult, and many patients with this disease are only diagnosed post mortem . There are few published reports of large series of patients with S aureus endocarditis and none from a community hospital . I reviewed the clinical and laboratory findings of a large number of patients with S aureus endocarditis in a community hospital . METHODS: I reviewed medical records identified through consultation records, International Classification of Diseases, Ninth Edition codes, and autopsy records of patients who fulfilled the criteria for the diagnosis of S aureus endocarditis during 1980 to 1991 . RESULTS: During the 12-year period, there were 106 cases, for a prevalence of 0.34 per 1000 admissions . Ninety-three (87.7%) of these patients were seen by me . The patients' ages ranged from 12 to 83 years (median, 61 years) . Eighteen cases were nosocomial (15 were associated with intravascular catheters) . Twenty-one patients were injecting drug users . Severe back pain was the chief complaint in nine patients . Twenty-seven patients had no heart murmur at the time of diagnosis . The overall mortality was 25.5% . CONCLUSIONS: Age 60 years or older, female gender, community-acquired infection, absence of heart murmur, presence of congestive heart failure, or central nervous system involvement was associated with higher mortality . Tricuspid valve endocarditis alone was associated with lower mortality.

Biochem J, 1994 Oct 15, 303 ( Pt 2), 633 - 8
Activation of the glycosyl-phosphatidylinositol-anchored membrane dipeptidase upon release from pig kidney membranes by phospholipase C; Brewis IA et al.; Incubation of pig kidney microvillar membranes with Bacillus thuringiensis or Staphylococcus aureus phosphatidylinositol-specific phospholipase C (PI-PLC) resulted in the release of a number of glycosyl-phosphatidylinositol (GPI)-anchored hydrolases, including alkaline phosphatase (EC 3.1.3.1), amino-peptidase P (EC 3.4.11.9), membrane dipeptidase (EC 3.4.13.19), 5'-nucleotidase (EC 3.1.3.5) and trehalase (EC 3.2.1.28) . Of these five ectoenzymes only for membrane dipeptidase was there a significant (approx . 100%) increase in enzymic activity upon release from the membrane . Maximal activation occurred at a PI-PLC concentration 10-fold less than that required for maximal release . In contrast solubilization of the membranes with n-octyl beta-D-glucopyranoside had no effect on the enzymic activity of membrane dipeptidase . A competitive e.l.i.s.a . with a polyclonal antiserum to membrane dipeptidase indicated that the increase in enzymic activity was not due to an increase in the amount of membrane dipeptidase protein . Although PI-PLC cleaved the GPI anchor of the affinity-purified amphipathic form of pig membrane dipeptidase there was no concurrent increase in enzymic activity . In the absence of PI-PLC, membrane dipeptidase in the microvillar membranes hydrolysed Gly-D-Phe with a Km of 0.77 mM and a Vmax . of 602 nmol/min per mg of protein . However, in the presence of a concentration of PI-PLC which caused maximal release from the membrane and maximal activation of membrane dipeptidase the Km was decreased to 0.07 mM while the Vmax . remained essentially unchanged at 624 nmol/min per mg of protein . Overall these results suggest that cleavage by PI-PLC of the GPI anchor on membrane dipeptidase may relax conformational constraints on the active site of the enzyme which exist when it is anchored in the lipid bilayer, thus resulting in an increase in the affinity of the active site for substrate.

Biochem J, 1994 Oct 15, 303 ( Pt 2), 517 - 25
The intracellular distribution of inositol polyphosphates in HL60 promyeloid cells; Stuart JA et al.; 1 . HL60 promyeloid cells contain high intracellular concentrations of inositol polyphosphates, notably inositol 1,3,4,5,6-pentakisphosphate (InsP5) and inositol hexakisphosphate (InsP6) . To determine their intracellular location(s), we studied the release of inositol (poly)phosphates, of ATP, and of cytosolic and granule-enclosed enzymes from cells permeabilized by four different methods . 2 . When cells were treated with digitonin, all of the inositol phosphates were released in parallel with the cytosolic constituents . Most of the InsP5 and InsP6 was released before significant permeabilization of azurophil granules . 3 . Similar results were obtained from cells preloaded with ethylene glycol and permeabilized by osmotic lysis . 4 . Electroporation at approximately 500 V/cm caused rapid release of free inositol . Higher field strengths provoked release of most of the ATP, InsP5 and InsP6, but only slight release of the intracellular enzymes . Multiple discharges released approximately 80-90% of total InsP5 and InsP6 . In the absence of bivalent-cation chelators, InsP5 and InsP6 were released less readily than ATP . 5 . Treatment of cells with Staphylococcus aureus alpha-toxin caused quantitative release of inositol and ATP, without release of intracellular enzymes . However, inositol phosphates were released much less readily than inositol or ATP . Even after prolonged incubation with a high concentration of alpha-toxin, only approximately 50-70% of InsP2, InsP3 and InsP4 and < or = 20% of InsP5 and InsP6 were released, indicating that the high charge or large hydrated radius of InsP5 and InsP6 might limit their release through small toxin-induced pores . 6 . These results indicate that most intracellular inositol metabolites are either in, or in rapid exchange with, the cytosolic compartment of HL60 cells . However, they leave open the possibility that a small proportion of cellular InsP5 and InsP6 (< or = 10-20%) might be in some intracellular bound form.

J Immunol, 1994 Oct 1, 153(7), 2974 - 82
Staphylococcal protein A induces biased production of Ig by VH3-expressing B lymphocytes; Kristiansen SV et al.; Staphylococcal protein A (SPA) is known to bind preferentially to the VH3 family of Ig heavy chain variable gene products . The current studies were conducted to examine the functional capacity of SPA to induce Ig production by VH3-expressing human B cells preferentially . Human peripheral blood B cells stimulated with anti-CD3-activated T cells or the Wood 46 strain of Staphylococcus aureus, which lacks SPA, expressed IgM containing all VH families, as detected by reverse transcription-PCR . In contrast, stimulation with SPA containing S . aureus or SPA-Sepharose resulted in biased expression of VH3 containing IgM . Similarly, cord blood B cells that require costimulation for Ig production induced by anti-CD3-activated T cells produced only VH3 containing IgM on costimulation with SPA containing S . aureus . The sequences of 21 VH3 IgM gene products derived from different B cell sources were determined and found to include at least nine members of the VH3 family, which were both in germ-line configuration and somatically mutated . Use of DH and JH was diverse . Analysis of these VH3 gene products revealed conserved residues in FR1 and 3'CDR2/FR3, which are candidates to play a role in SPA-mediated activation of VH3-expressing B cells . Like T cell superantigens, SPA probably interacts with residues in the partially solvent exposed regions of the heavy chain molecule outside the classical Ag binding site and provides a signal that can lead to Ig production by human B cells expressing VH3.

Microbiology, 1994 Oct, 140 ( Pt 10), 2577 - 83
An investigation of plasmids from Staphylococcus aureus that mediate resistance to mupirocin and tetracycline; Needham C et al.; Plasmids conferring mupirocin resistance were prepared from isolates of Staphylococcus aureus obtained from four patients in the same ward . The plasmids are related and in all of them the gene conferring mupirocin resistance (mupA) is flanked by copies of IS257 in direct repeat . In two plasmids mupA and IS257 have been duplicated and in one of these plasmids (pJ3358) a small pT181-like plasmid conferring tetracycline resistance is present flanked by copies of IS257 . Filter mating with a strain containing pJ3358 as donor and selection on tetracycline sometimes resulted in transfer of the pT181-like plasmid containing a copy of IS257 . Analysis showed that the pT181-like plasmid with the insertion of IS257 is present in high copy number and that the IS257 element is inserted in the copy number control region of the plasmid.

Rinsho Byori, 1994 Oct, 42(10), 1069 - 76
{Detection of methicillin-resistant Staphylococcus aureus using PCR and non-radioactive DNA probes: III . Mutations of the fem A gene in clinical strains of Staphylococcus aureus}; Yamashita K et al.; Methicillin resistance in S . aureus is primarily due to the presence of the mec A gene . However, in addition to mec, the phenotypic expression of methicillin resistance requires the presence of an additional gene(s), fem A which is chromosomally encoded . Previous studies suggest an increase in the biochemical function of fem A gene products due to base substitutions in the region upstream of the fem A gene and in its coding frame . The partial nucleotide sequences of fem A regions in reference and clinical strains of S.aureus were therefore analyzed by PCR-direct solid-phase sequencing and suitable DNA probes . Amplified target DNAs of 251, 330 and 271 bp were resolved on ethidium bromide-stained gels and hybridized with DNA probes conjugated to alkaline phosphatase . In ATCC 12600 strain, a palindromic sequence was conserved in the region upstream of fem A . However, it was destroyed by the occurrence of mutations in other reference, and clinical strains tested regardless of whether they are methicillin-susceptible or resistant . Furthermore, in the coding frame of fem A, two missense mutations were present in MSSA and MRSA without any regularity . These findings suggest that mutations in the fem A region may not be a single factor essential for regulation of methicillin resistance, although fem A probably functions cooperatively with mec A.

J Orthop Trauma, 1994 Oct, 8(5), 383 - 9
Distribution of clindamycin in cortical bone during direct local infusion of the canine tibia; Budsberg SC et al.; The distribution of clindamycin in tibial cortical bone, administered via direct local infusion with an implantable pump, is described . Clindamycin concentrations in cortical bone were measured after 3, 7, and 21 days of intraosseous infusion . The tibia were divided into four quadrants relative to the outflow infusion catheter site located in the medial aspect of the mid-diaphysis . A gradient of 5-30 mm from the infusion site was documented in all four quadrants (proximal lateral, proximal medial, distal lateral, and distal medial) . At all sampling times, clindamycin concentrations in all quadrants exceeded the minimum inhibitory concentrations for gram-positive aerobic bacteria, including Staphylococcus aureus and S . epidermidis, and both the gram-positive and gram-negative anaerobes, including Peptostreptococcus species and Bacteroides species . The data suggest that gravitational forces affect the diffusion of the clindamycin because concentrations in both distal quadrants were greater than in corresponding proximal quadrants.

Kansenshogaku Zasshi, 1994 Oct, 68(10), 1211 - 7
{The improvement on the nursing procedure for pregnant women and newborns and the decrease of Staphylococcus aureus infections in newborns and infants in our hospital}; Hamada T; Knowing the importance of prevention of Staphylococcus aureus infections, especially that of Methicillin-resistant S . aureus (herein after abbreviated as MRSA) in the newborn room, we improved the nursing procedure in the room and examined subsequent changes in the onset condition of the above infection in newborns and infants less than 1-year-old . The infection was detected in 13 cases of stage 1 (stage according to the nursing procedure since the opening of our hospital): 6 cases (all MRSA) had their onset in the room . MRSA was detected in 11 cases, suspecting a hospital cross infection in relation to the period of birth and coagulase type . The infection was detected in 9 cases of stage 2 (stage of prevention of inter-newborn hospital cross infection through environmental purification within the newborn room, through hand and finger disinfection of medical workers, through improved bathing, etc.); 2 cases (1 of MRSA, 1 of Methicillin-sensitive S . aureus {hereinafter abbreviated as MSSA}) had an onset in the room . MRSA was detected in 5 cases, but hospital cross infection was thought difficult to be responsible . The MRSA infection was detected in 4 pregnant cases (0.8%) of stage 3 (stage of effortful prevention of MRSA from carrying into the ward following examination of possible intra-nasal MRSA establishment in pregnant women in addition to similar newborn nursing to stage 2), indicating pregnant women's possible carrying MRSA into the newborn room . S . aureus infection was detected in 9 cases; 2 cases (MSSA) had an onset in the above room.(ABSTRACT TRUNCATED AT 250 WORDS)

Med Clin (Barc), 1994 Oct 1, 103(10), 361 - 5
{Application of molecular epidemiology techniques in the study of food poisoning caused by Staphylococcus aureus}; Montserrat I et al.; BACKGROUND: The use of molecular epidemiology techniques has provided better knowledge as to the clonal organization of bacterian populations and thus allows better follow up of epidemics . An alimentary toxiinfection in a Barcelona school produced by Staphylococcus aureus was analyzed by the combination of epidemiologic, phenotype and genotype markers with the aim of determining the source of the alimentary contamination . METHODS: Nine strains of Staphylococcus aureus isolated in 6 food manipulators and 3 patients were studied with the following markers: biotype, antibiotype, phagotype, plasmid profile, polymorphism of the size of the restriction fragments of total DNA and ribotype . RESULTS: Epidemiologic study of the strains analyzed showed that both the phenotype markers and the plasmid profile are thecniques of little discriminatory value . The only clearly discriminatory technique used was ribotyping which defined 3 clones in the 9 strains of Staphylococcus aureus studied . CONCLUSIONS: Molecular study of isolated strains of Staphylococcus aureus was able to identify the causal origin of the alimentary toxiinfection in one of the 6 food manipulators studied.

Enferm Infecc Microbiol Clin, 1994 Oct, 12(8), 393 - 7
{Tricuspid endocarditis caused by penicillin-resistant Staphylococcus aureus: 2 cases in parenteral drug addicts}; Dronda F et al.; BACKGROUND: Staphylococcus aureus is the main causal agent of infectious endocarditis (IE) in intravenous drug addicts (IVDA) with most of the strains, isolated in Spain being resistant to penicillin and sensitive to methycillin, although the latter condition varied in recent years . Two cases of IE caused by S . aureus strains sensitive to penicillin in IVDA are presented . METHODS: All bacteremia episodes diagnosed in the Hospital General Penitenciario (Madrid) over a 33-month period (March 1991 to December 1993) were prospectively studied . Special attention was given to patients diagnosed of IE . Blood cultures were processed according to the usual technique by a non radiometric system . RESULTS: One hundred and four bacteremias were detected with 14 being produced by S . aureus . Ten episodes of the total number of bacteremias fulfilled criteria for IE with 2 being produced by strains of S . aureus sensitive to penicillin (CMI < 0.06 mu/ml) . Both patients had coinfection by HIV . The clinical evolution prior to diagnosis was prolonged although clinical and microbiologic cure were achieved with intravenous beta-lactam antibiotics treatment, without complications . CONCLUSIONS: Despite the low incidence of the isolation of Staphylococcus aureus sensitive to penicillin (< 3%) recently observed in Spain, strains producing severe infections, showing patterns of sensitivity such as those found during the preantibiotic era, may still be isolated.

No Shinkei Geka, 1994 Oct, 22(10), 973 - 6
{Cervical spinal epidural abscess caused by methicillin-resistant Staphylococcus aureus (MRSA)}; Sugimoto M et al.; Epidural abscess is an uncommon infectious disease . The cervical spine is the least frequent site of spinal epidural abscess . It has been reported that early diagnosis and surgical treatment prevent neurological deficit, but it is difficult to diagnose this disease clinically . We presented a rare case of cervical epidural abscess caused by MRSA . A 54-year-old man was admitted to our hospital because of acute renal failure and hepatitis . He was treated with hemodialysis via the femoral route . His renal function recovered but high fever continued . MRSA was identified from the AV shunt catheter . He noted pain and dysesthesia on his left shoulder one month after admission . He was transferred to our department with suspect of spinal tumor . Neurological examination demonstrated left hemiparesis with superficial sensory disturbance between C8 and Th2 . Cervical CT scan showed osteomyelitis at the left C7 lamina and facet . MR imaging disclosed that an epidural mass at C7 had low signal intensity on T1 weighted and high signal intensity on T2 weighted and ring-like enhancement with gadolinium . He was treated conservatively for a month . Sequential MR imaging showed the mass had homogeneous enhancement at C7 epidural space extending to the left intervertebral foramen . Laboratory examination showed normal . The patient was diagnosed as having cervical epidural abscess . A C6 through Th2 laminectomy and C8 foraminotomy were performed and an encapsulated abscess including yellowish pus was totally removed . The pathological diagnosis was non-specific abscess in the subacute stage . MRSA was identified by the intraoperative pus culture . After the surgery, antibiotics were administered.(ABSTRACT TRUNCATED AT 250 WORDS)

Br J Dermatol, 1994 Oct, 131(4), 536 - 40
Local steroid therapy and bacterial skin flora in atopic dermatitis; Stalder JF et al.; A double-blind, randomized trial was conducted to determine the influence of topical steroid therapy on atopic skin flora . The bacteriological and clinical effects of desonide (Locapred), compared with those of its excipient, were studied in 40 children . Clinical scoring and bacteriological sampling were performed before the start of the trial and after 7 days of once-daily topical treatment . Before treatment, no differences in clinical score or Staphylococcus aureus colonization were noted between the two groups . After treatment, the clinical score improved (P < 0.001) in the desonide group, and S . aureus density decreased dramatically (P < 0.001) . In the excipient group, no significant differences in clinical score or S . aureus density were noted . A comparison of the two groups demonstrated statistically significant differences with regard to clinical score (P < 0.001) and S . aureus density (P < 0.05) . These results show the efficacy of topical corticosteroid treatment alone on S . aureus colonization in atopic skin, and confirm the critical role of inflammation in bacterial colonization.

Arch Pathol Lab Med, 1994 Oct, 118(10), 1023 - 5
Isolated myocardial abscess causing coronary artery rupture and fatal hemopericardium; Fan CC et al.; We report a rare case of cardiac tamponade caused by hemopericardium secondary to erosion and rupture of a coronary artery by an adjacent solitary myocardial abscess . The resulting tamponade led to the sudden death of this 46-year-old man . Antemortem blood culture and the postmortem microscopic examination of the myocardial abscess revealed the causative agent to be Staphylococcus aureus . Hemopericardium due to ventricular wall rupture secondary to a myocardial abscess has been infrequently reported, but, to our knowledge, only one other report of hemopericardium due to coronary artery rupture related to myocardial abscess has been published.

Am J Med Sci, 1994 Oct, 308(4), 251 - 4
Case report: diabetes mellitus as a predisposing factor in the development of pyomyositis; Belsky DS et al.; Pyomyositis is an uncommon infection in temperate climates, usually resulting from Staphylococcus aureus infection of skeletal muscle . In this report, the authors describe a patient with untreated Type 2 diabetes mellitus who suffered nonpenetrating blunt trauma to his left anterior thigh, and S . aureus pyomyositis and secondary osteomyelitis of his proximal tibia and patella subsequently developed as a result of delayed diagnosis and treatment . Patients with diabetes mellitus are at increased risk for the development of pyomyositis because of more frequent S . aureus colonization of skin, nasal mucosa, and oropharynx; a delay in definitive treatment can lead to significant morbidity in these patients . Computed tomography or magnetic resonance imaging may be helpful in the diagnosis of pyomyositis . An anemia of chronic disease may result from this disorder, which resolves with treatment.

J Surg Res, 1994 Oct, 57(4), 505 - 9
Gut ischemia induces bone marrow failure and increases risk of infection; Fontes B et al.; Hemorrhagic shock leads to bone marrow (BM) failure and renders the host susceptible to infection . We hypothesized that splanchnic hypoperfusion may play a mechanistic role in this process . BM was harvested from normal rats and, on Postprocedure Days 1 and 3, from rats that had undergone laparotomy (LAP) or gut ischemia/reperfusion (I/R; 45 min superior mesentery artery occlusion) . Granulocyte-macrophage colony-forming unit (CFU-GM) proliferation, a measure of BM myeloid progenitors, was quantitated using a standard soft agar culture technique . On Postprocedure Days 1 and 3, BM proliferation of CFU-GM was depressed in gut I/R rats, compared to control and LAP animals (P < 0.05) . Next, six rats were subjected to I/R, LAP, ANEST (anesthesia control), or no treatment (NL, normal control); 1 day later, 3.5 x 10(7) Staphylococcus aureus, suspended in 0.25 ml of saline, were injected subcutaneously in four sites on the back of each animal . Five days later, the NL rats had developed 23 abscesses, ANEST 23, and LAP 22, while the gut I/R rats had 24 . The abscesses were excised, weighed, and measured . The weight and size of abscesses were greater in the gut I/R animals (P < 0.05) . In summary, gut I/R depressed BM proliferation and rendered animals susceptible to infection in a manner similar to that observed following hemorrhagic shock . These data suggest that splanchnic hypoperfusion, a common sequela of hemorrhagic shock, may play a mechanistic role in BM failure and infection after hemorrhage.

J Med Microbiol, 1994 Oct, 41(4), 282 - 90
Detection by polymerase chain reaction of genes encoding aminoglycoside-modifying enzymes in methicillin-resistant Staphylococcus aureus isolates of epidemic phage types . Belgian Study Group of Hospital Infections (GDEPIH/GOSPIZ); Vanhoof R et al.; The polymerase chain reaction (PCR) was used to identify the aacA-aphD, aphA3 and aadC genes, encoding the aminoglycoside-modifying enzymes AAC(6')-APH(2"), APH(3')III and ANT(4'4"), respectively, and the methicillin resistance determinant mecA, in epidemic aminoglycoside and methicillin-resistant isolates of Staphylococcus aureus . In total, 37 isolates collected in the period 1980-1985 and 81 isolates from the period 1991-1992 were obtained from 10 different Belgian hospitals . Epidemic isolates from the earlier period were characterised by phage type C (6/47/54/75) of phage group III, whereas two other epidemic phage types of group III-types A (77) and B (47/54/75/77/84/85)--were commonest in isolates from the second period . The bifunctional AAC(6')-APH(2") was the enzyme encountered most frequently . The prevalence of APH(3')III decreased significantly in the 1991-1992 period, while ANT(4',4") was found solely in isolates from this period . Resistance mechanisms were more complex in isolates from the 1991-1992 period and the mecA gene was detected in all isolates . The PCR results corresponded well with those obtained in the radiochemical phosphocellulose paper binding assay . Isolates from the 1991-1992 period were shown to express significantly higher levels of acetyltransferase activity than isolates from the 1980s.

J Med Microbiol, 1994 Oct, 41(4), 259 - 63
Virulence of coagulase-deficient mutants of Staphylococcus aureus in experimental endocarditis; Baddour LM et al.; A sensitive rat endocarditis model which employed relatively small inocula (< or = 10(4) cfu) was used to examine the role of coagulase in the pathogenesis of infection . Rats with indwelling, intracardiac catheters were challenged intravenously with three strains of Staphylococcus aureus . The virulence of a coagulase-positive parental strain DU5808 was compared in terms of its ID50 for resected vegetations and catheters to that of two coagulase-negative mutant strains (DU5809 and DU5814) which had undergone site-specific mutagenesis . Confidence intervals of infection rates were calculated and comparisons were performed of weights of infected vegetations, bacterial concentrations in vegetations and early mortality rates . From these virulence parameters, it was concluded that there were no differences in virulence among the three strains . The results from this investigation support the previous findings in mouse models of subcutaneous infection and mastitis, which indicated that coagulase production by S . aureus does not appear to function as a virulence factor . Together, these data refute the longheld belief that coagulase is important in the pathogenesis of infection and indicate that other markers of virulence must be operative in diseases caused by the enduring pathogen S . aureus.

J Parasitol, 1994 Oct, 80(5), 756 - 63
Purification and characterization of surface-associated proteins from adult Haemonchus contortus; Rhoads ML et al.; Extrinsic radioiodination experiments have shown that male and female adults of Haemonchus contortus (BPL strain) express a stage-specific set of surface-associated proteins with apparent molecular mass values of 30, 58, 81, and 143 kDa . A quantitatively different pattern of iodinated surface proteins is expressed by adults of the PPR strain of H . contortus, whereas the pattern of iodinated proteins expressed by Haemonchus similis is qualitatively distinct (38, 68, and 121 kDa) . The 58-, 81-, and 143-kDa proteins of the BPL strain are glycosylated, whereas the 30-kDa protein is not . The binding of wheat germ agglutinin to the surface glycoproteins was inhibited by the trimer of N-acetylglucosamine (N,N,N-triacetylchitotroise) but not by the monosaccharide, indicating the presence of chitin-like homopolymers . The carbohydrate portion of the 58-kDa protein is N-linked and accounts for 30% of its apparent mass . Under nonreducing conditions, the 58-kDa glycoprotein forms a high molecular mass polymer that is unable to penetrate a 10% acrylamide gel . The 143- and 81-kDa surface glycoproteins were not hydrolyzed by either N- or O-glycanase, indicating unusual modifications to the saccharide-linkage and rendering it resistant to glycosidase digestion . The 30-, 58-, and 143-kDa purified surface proteins produced distinct peptide maps with Staphylococcus aureus V8 protease.

J Infect Dis, 1994 Oct, 170(4), 818 - 27
Toxic shock syndrome toxin 1-producing Staphylococcus aureus isolates contain the staphylococcal enterotoxin B genetic element but do not express staphylococcal enterotoxin B; De Boer ML et al.; Toxic shock syndrome toxin 1 (TSST-1) and staphylococcal enterotoxin B (SEB) are two examples of variable genetic traits produced by Staphylococcus aureus . Regarding phenotypes, most investigations have reported that SEB and TSST-1 are not coproduced in S . aureus . However, it is unclear whether genotypically SEB (entB) and TSST-1 (tst) genes are found within the same isolate . Gene probes specific for entB and tst showed that both genes were present within 8 TSST-1+ isolates analyzed . However, SEB was not produced by any of these isolates . In contrast, none of 9 SEB-producing S . aureus isolates contained the TSST-1 genetic element DNA . Hybridization of restriction fragments with TSST-1 and SEB genetic element probes and tryptophan auxotyping suggested that tst and entB are located in close proximity on the chromosome . The entB locus may be a preferred site of insertion for the TSST-1 genetic element, and such insertion may interfere with the expression of SEB.

J Infect Dis, 1994 Oct, 170(4), 1033 - 7
Gentamicin-resistant menadione and hemin auxotrophic Staphylococcus aureus persist within cultured endothelial cells; Balwit JM et al.; Staphylococcus aureus menadione and hemin auxotrophs, generated by in vitro gentamicin selection, demonstrated reduced hemolytic activity and enhanced intracellular survival within cultured bovine aortic endothelial cells relative to their hemolytic parent . Supplementation of the auxotrophs with exogenous menadione or hemin resulted in rapid growth, increased hemolytic activity, and reduced intracellular persistence to the level found for the hemolytic clinical parent . Aminoglycoside selection of staphylococcal menadione and hemin auxotrophs and subsequent persistence of these variants in the intracellular milieu may adapt S . aureus for evasion of host defenses and resistance to antimicrobial therapy.

J Bone Joint Surg Am, 1994 Oct, 76(10), 1500 - 6
Polylactide/polyglycolide antibiotic implants in the treatment of osteomyelitis . A canine model; Garvin KL et al.; Osteomyelitis with Staphylococcus aureus was established in the tibiae of twenty-six adult mongrel dogs . After confirmation of infection at four weeks, all animals had operative debridement and were then divided into three treatment groups . Group 1 (eight animals {sixteen tibiae}) was treated with parenteral administration of gentamicin (three milligrams per kilogram of body weight per day) every eight hours for four weeks . Group 2 (nine animals {nine tibiae}) was treated with a polymethylmethacrylate implant containing 100 milligrams of gentamicin that was placed in the tibia for six weeks . Group 3 (nine animals {nine tibiae}) was treated with a polylactide/polyglycolide implant containing 100 milligrams of gentamicin that was placed in the tibia for six weeks . All animals were killed at the end of treatment . At that time, specimens of tissue were obtained for quantitative culture as well as for antibiotic immunoassay . In the groups that had been treated with an implant, serum was obtained for the measurement of serum drug levels after debridement; after the implantation; four, seven, and twenty-one days postoperatively; and immediately before the animals were killed . The infection was eradicated in ten of the sixteen tibiae in Group 1, in eight of the nine tibiae in Group 2, and in all nine tibiae in Group 3.(ABSTRACT TRUNCATED AT 250 WORDS)

Infect Immun, 1994 Oct, 62(10), 4637 - 40
Protective role of sialophorin (CD43)-expressing cells in experimental Staphylococcus aureus infection; Bremell T et al.; Sialophorin (CD43) is a major surface mucin on most hematopoietic cell lineages, including phagocytes . Defects of CD43 expression occur in Wiskott-Aldrich syndrome, a disease characterized by susceptibility to pyogenic infections . In a newly established rat model of septic Staphylococcus aureus arthritis, we have investigated the role of CD43-expressing cells in the progression of the disease . A single injection of a monoclonal antibody specific for CD43 induced a highly erosive course of arthritis and increased mortality in animals exposed to a suboptimal dose of bacteria . Our results demonstrate that sialophorin-expressing cells play a protective role in the early stage of staphylococcal infection.

Infect Immun, 1994 Oct, 62(10), 4556 - 63
Staphylococcus saprophyticus hemagglutinin binds fibronectin; Gatermann S et al.; Attachment of microorganisms to host tissue is regarded as an important step in the pathogenesis of infections . Staphylococcus saprophyticus adheres to various epithelial cells and hemagglutinates sheep erythrocytes . The hemagglutinin has been identified, but a human target for this surface protein is still not known . In our report, we show that hemagglutinating strains of S . saprophyticus bind to immobilized fibronectin, whereas nonhemagglutinating strains do not . Bacterial binding was inhibited by antibody to the hemagglutinin but not by antibody to Ssp, another surface protein of S . saprophyticus . The purified hemagglutinin but not other surface proteins bound biotin-labeled fibronectin . Binding was saturable and could be inhibited by unbound hemagglutinin, unlabeled fibronectin, and by antibody to the hemagglutinin . We thus conclude that the hemagglutinin of S . saprophyticus may act as a fibronectin receptor in the human host . Heparin, the D3 peptide, or Arg-Gly-Asp-Ser (RGDS) containing peptides did not inhibit binding of fibronectin to the hemagglutinin, indicating that the binding site is different from that of Staphylococcus aureus or Treponema pallidum.

Infect Immun, 1994 Oct, 62(10), 4304 - 9
Effect of genetic variation on induced neutrophilia in mice; Marley SB et al.; Mice from a variety of strains were injected with a sterile irritant (Brewer's thioglycolate) and killed bacteria (Staphylococcus aureus, Staphylococcus epidermidis, or Escherichia coli) to determine their effect on accumulation of neutrophils in the peritoneal cavity . Peak accumulation occurred around 15 h postinjection and showed significant strain-related variation . C57BL/10 mice were identified as having a high-responder phenotype and BALB/c mice a low-responder phenotype . Inheritance of the high-responder phenotype followed simple Mendelian genetics: (BALB/c x C57BL/10)F1 mice were found to be more responsive than either parental phenotype . Major histocompatibility complex H-2d haplotype was found to convey an augmented neutrophil response in conjunction with B10 background high-responder genes (B10.D2/n) but the H-2d haplotype per se was not the only factor in determining high responsiveness . Gram-positive and gram-negative bacteria appeared to activate different immune mechanisms . Both gram-negative bacteria and lipopolysaccharides (LPS) induced a response similar to, but less potent than, that induced by Brewer's thioglycollate . Neutralization of the LPS content of Brewer's thioglycolate abrogated the response.

Infect Immun, 1994 Oct, 62(10), 4296 - 303
Protein tyrosine kinase activity is essential for Fc gamma receptor-mediated intracellular killing of Staphylococcus aureus by human monocytes; Zheng L et al.; Our previous study revealed that the intracellular killing of Staphylococcus aureus by human monocytes after cross-linking Fc gamma receptor I (Fc gamma RI) or Fc gamma RII is a phospholipase C (PLC)-dependent process . The aim of the present study was to investigate whether protein tyrosine kinase (PTK) activity plays a role in the Fc gamma R-mediated intracellular killing of bacteria and activation of PLC in these cells . The results showed that phagocytosis of bacteria by monocytes was not affected by the PTK inhibitors genistein and tyrphostin-47 . The intracellular killing of S . aureus by monocytes after cross-linking Fc gamma RII or Fc gamma RII with anti-Fc gamma R monoclonal antibody and a bridging antibody or with human immunoglobulin G (IgG) was inhibited by these compounds in a dose-dependent fashion . The production of O2- by monocytes after stimulation with IgG or IgG-opsonized S . aureus was almost completely blocked by the PTK inhibitor . These results indicate that inhibition of PTK impairs the oxygen-dependent bactericidal mechanisms of monocytes . Genistein and tyrphostin-47, which do not affect the enzymatic activity of purified PLC, prevented activation of PLC after cross-linking Fc gamma RI or Fc gamma RII, measured as an increase in the intracellular inositol 1,4,5-trisphosphate concentration . Cross-linking Fc gamma RI or Fc gamma RII induced rapid tyrosine phosphorylation of several proteins in monocytes, one of which was identified as PLC-gamma 1, and the phosphorylation could be completely blocked by PTK inhibitors, leading to the conclusion that activation of PLC after cross-linking Fc gamma R in monocytes is regulated by PTK activity . Together, these results demonstrate that PTK activity is essential for the activation of PLC which is involved in the Fc gamma R-mediated intracellular killing of S . aureus by human monocytes.

Clin Immunol Immunopathol, 1994 Oct, 73(1), 96 - 102
Histamine inhibits immunoglobulin production via histamine H2 receptors without affecting cell growth in human B cells; Fujimoto M et al.; The effect of histamine upon immunoglobulin (Ig) production and proliferation in human B cells was studied . Histamine inhibited Ig production by the human B cell lines IM-9 and CBL in a dose-dependent fashion during 4 days of culture . As little as 10(-5) M was inhibitory . In contrast, proliferation was not affected . Inhibition by histamine was blocked by histamine H2 antagonist, cimetidine, but not by histamine H1 antagonist, diphenhydramine . Moreover, histamine H2 agonist dimaprit inhibited Ig production from B cell lines in a dose-dependent manner, and as little as 10(-8) M was inhibitory . Histamine also inhibited IgM and IgG production by peripheral and tonsillar B cells stimulated with Staphylococcus aureus Cowan strain I and IL-2 without affecting proliferation . This inhibition was also blocked by cimetidine . These results indicate that histamine has a direct inhibitory effect on B cells via histamine H2 receptors, and acts as an immunoregulatory factor.

Aging (Milano), 1994 Oct, 6(5), 368 - 71
Methicillin-resistant Staphylococcus aureus colonization in a new nursing home; Feingold K et al.; Methicillin-resistant Staphylococcus aureus (MRSA) has been detected in nursing homes and long-term care facilities . Studies disagree about the risk of infection with MRSA in colonized patients . MRSA colonization and infection were tracked for one year in all admissions to a 60-bed ward at the Philadelphia VA Nursing Home Care Unit (NHCU) from the time of its opening in June, 1990 . Patients and staff were blinded to culture results, and the NHCU followed universal precautions for all patients . Of the first 72 patients, 7 were found to be colonized with MRSA; only one of them was known to have had MRSA prior to NHCU transfer . Three patients died (2 had negative cultures prior to death), and 1 was discharged home . Three patients spontaneously cleared MRSA colonization and lived to the end of the study . Three patients appeared to be colonized by MRSA after admission; subsequent cultures were negative . No patients were infected by MRSA in the NHCU . At the close of the study, one year after the nursing home opened, no patient in the nursing home had a culture positive for MRSA . In conclusion, colonization with MRSA at the time of admission to the nursing home is not uncommon, but patients can spontaneously clear it . Besides, nursing homes that pre-screen only those patients with classic risk factors may be admitting many MRSA-colonized patients . Nonetheless, universal precautions appear to be effective in limiting transmission of MRSA in the nursing home; in this study, MRSA acquisition was sporadic and brief.

Int J Food Microbiol, 1994 Oct, 23(2), 167 - 78
Identification of psychrotrophic Micrococcaceae spp . isolated from fresh beef stored under carbon dioxide or vacuum; Venugopal RJ et al.; Seventy-four Gram-positive, catalase-positive coccal strains were isolated from fresh beef stored under carbon dioxide (< 500 ppm O2) or vacuum for up to 15 weeks at 0, 2 or 4 degrees C . Isolates were identified using biochemical tests listed in several published protocols and the API Staph-Ident System . No isolates were identified as Staphylococcus aureus . Twenty-nine isolates were identified as Staphylococcus saprophyticus (five distinct groups), 24 isolates were identified as Staphylococcus gallinarum and 21 isolates were identified as Micrococcus varians . The staphylococcal isolates were coagulase-negative, non-hemolytic and novobiocin resistant . They produced acid from several carbohydrates under aerobic conditions, hydrolysed gelatin but not collagen, showed lipolytic activity and grew in 15% NaCl . The Micrococcus varians isolates also were salt-tolerant, produced acid only from glucose, fructose and galactose (two strains), and were resistant to lysozyme (1600 micrograms/ml) . Lactic acid was the major end product of aerobic glucose metabolism . All S . saprophyticus and M . varians isolates tested contained cell wall fatty acids with chain length > or = C20:0.

J Hosp Infect, 1994 Oct, 28(2), 113 - 26
Characterization of two different clusters of clonally related methicillin-resistant Staphylococcus aureus strains by conventional and molecular typing; Trzcinski K et al.; The DNA fragments of 28 distinct isolates of methicillin-resistant Staphylococcus aureus (MRSA) originating from different hospitals in Warsaw and Lodz, were studied . They were obtained by cleavage with restriction endonuclease SmaI and subsequently analysed by pulsed-field electrophoresis . Sixteen different patterns were seen and clusters of related strains were clearly distinguishable . Minor differences in fragment patterns within these clusters and among epidemiologically related strains, revealed genomic rearrangements in the course of clonal dissemination of particular strains . The isolates were also checked for the expression of methicillin resistance . Isolates with heterogenous and homogeneous phenotypes, fell into clearly distinct clusters and thus formed two clonally related MRSA strains . Differences were also seen with phage and biochemical typing, and antimicrobial resistance patterns.

Infect Control Hosp Epidemiol, 1994 Oct, 15(10), 646 - 51
Methicillin-resistant Staphylococcus aureus in tertiary care institutions on the Canadian prairies 1990-1992; Embil J et al.; OBJECTIVE: To review experience with methicillin-resistant Staphylococcus aureus (MRSA) in tertiary acute-care teaching hospitals on the Canadian prairies . DESIGN: Retrospective review for a 36-month period, 1990 through 1992 . SETTING: Five tertiary acute-care teaching hospitals in three Canadian prairie provinces . METHODS: MRSA isolates and susceptibility were identified through the clinical microbiology laboratory at each institution . For each patient, data collected included duration of institutional residence prior to isolation, patient ethnic background, age, sex, and antimicrobial susceptibility . Epidemiologic typing of strains used restriction fragment length polymorphism analysis by pulsed-field gel electrophoresis . RESULTS: Two hundred fifty-nine MRSA isolates were identified in 135 patients during the 36 months, with substantial institutional variation in number of isolates . No consistent increase in yearly numbers of isolates was apparent . Patients usually had MRSA identified at admission (62%); only one of five centers had the majority of isolates acquired nosocomially . Patients with MRSA present at admission were more frequently of aboriginal (First Nations) ethnicity (62% compared with 14% of nosocomial; P < 0.001) . Pulsed-field gel electrophoresis of 167 isolates from 135 patients revealed 46 different strains with little interprovincial or interinstitutional identity of strains . CONCLUSIONS: MRSA isolated in patients in tertiary care institutions in these three Canadian provinces usually is acquired prior to admission . A disproportionate number of isolates are identified in aboriginal Canadians . Epidemiologic typing was consistent with a polyclonal origin of MRSA in this geographic area.

Antimicrob Agents Chemother, 1994 Oct, 38(10), 2480 - 2
Efficacies of different vancomycin dosing regimens against Staphylococcus aureus determined with a dynamic in vitro model; Duffull SB et al.; A dynamic in vitro model was used to assess four different vancomycin dosing regimens against Staphylococcus aureus . These regimens achieved peak drug concentrations of 48 micrograms/ml (single dose) and 30 micrograms/ml (dosed every 12 h) and constant concentrations of 16 and 8 micrograms/ml . Analysis of the area under the bacterial concentration-time curve, area under the first moment of the bacterial concentration-time curve, and bacterial elimination rate constant showed no difference in the rate or extent of bacterial killing . The optimal dosing method may be that which achieves the lowest area under the curve while concentrations are maintained above the MBC.

Antimicrob Agents Chemother, 1994 Oct, 38(10), 2426 - 9
Intracellular penetration and activity of BAY Y 3118 in human polymorphonuclear leukocytes; Garcia I et al.; The penetration of a new quinolone (BAY Y 3118) into human polymorphonuclear leukocytes (PMNs) was evaluated by a fluorometric assay . The cellular concentration-to-extracellular concentration (C/E) ratio was higher than 6.3 at extracellular concentrations ranging from 2 to 100 mg/liter . The uptake of BAY Y 3118 was rapid, reversible and nonsaturable . The intracellular penetration of BAY Y 3118 was significantly affected by environmental temperature (C/E ratio at 4 degrees C, 5.4 +/- 0.5; control, 7.5 +/- 0.9; P < 0.05) and cell viability (C/E ratio in dead PMNs, 5.5 +/- 0.8; control 7.5 +/- 0.9; P < 0.05), but it was not affected by metabolic inhibitors . The ingestion of opsonized zymosan or opsonized Staphylococcus aureus significantly decreased the levels of PMN-associated BAY Y 3118 . Cell stimulation by a membrane activator, however, significantly increased the intracellular concentration of this quinolone . At therapeutic extracellular concentrations (0.5, 2, and 5 mg/liter), BAY Y 3118 showed intracellular activity greater than that of ciprofloxacin against S . aureus in human PMNs . It was concluded that BAY Y 3118 reaches high intracellular concentrations within human PMNs and remains active intracellularly.

J Dairy Sci, 1994 Oct, 77(10), 2965 - 74
Effect of whole Staphylococcus aureus and mode of immunization on bovine opsonizing antibodies to capsule; Guidry AJ et al.; Exopolysaccharide capsule is a major virulence factor of Staphylococcus aureus because it inhibits neutrophil recognition of antibodies to highly antigenic S . aureus cell wall . To circumvent this inhibition, two modes of immunization were tested for ability to induce anticapsular opsonins . Cows were immunized at drying off and boosted on d 14 and 28 by injection of Smith diffuse S . aureus plus dextran sulfate in the area of the supramammary lymph node or intramammarily . In cows immunized in the area of the supramammary lymph node, IgG1 and IgG2 sera antibody titers to capsule increased and remained elevated to the end of the study, 120 d postcalving . The IgM titers increased during the dry period but declined to preimmunization levels at calving . Response of serum IgG1 and IgM to intramammary immunization was similar to that with supramammary lymph node immunization, but more delayed and lower in magnitude . Antibodies of all four isotypes, IgG1, IgG2, IgA, and IgM, increased in dry secretions following immunization via lymph node . In cows immunized in the lymph node, IgG1 antibodies remained elevated throughout the study, but IgG2 antibodies dropped to baseline 15 d postcalving . In cows immunized intramammarily, only IgA antibodies increased significantly in lacteal secretions and remained elevated throughout the study . Immunization of cows in the lymph node during the dry period enhanced the ability of dry secretions and colostrum to support phagocytosis.

Br Heart J, 1994 Oct, 72(4), 339 - 43
Antibiotic prophylaxis in permanent pacemaker implantation: a prospective randomised trial; Mounsey JP et al.; BACKGROUND--Pacemaker pocket infection is a potentially serious problem after permanent pacemaker implantation . Antibiotic prophylaxis is commonly prescribed to reduce the incidence of this complication, but current trial evidence of its efficacy is conflicting . A large prospective randomised trial was therefore performed of antibiotic prophylaxis in permanent pacemaker implantation . The intention was firstly to determine whether antibiotic prophylaxis is efficacious in these patients and secondly to identify which patients are at the highest risk of infection . METHODS--A prospective randomised open trial of flucloxacillin (clindamycin if the patient was allergic to penicillin) v no antibiotic was performed in a cohort of patients undergoing first implantation of a permanent pacing system over a 17 month period . Intravenous antibiotics were started at the time of implantation and continued for 48 hours . The trial endpoint was a repeat operation for an infective complication . RESULTS--473 patients were entered into a randomised trial . 224 received antibiotic prophylaxis and 249 received no antibiotics . A further 183 patients were not randomised but were treated according to the operator's preference (64 antibiotics, 119 no antibiotics); these patients are included only in the analysis of predictors of infection . Patients were followed up for a mean (SD) of 19(5) months . Among the patients in the randomised group there were nine infections requiring a repeat operation, all in the group not receiving antibiotic (P = 0.003) . In the total patient cohort there were 13 infections, all but one in the non-antibiotic group (P = 0.006) . Nine of the infections presented as erosion of the pulse generator or electrode, three as septicaemia secondary to Staphylococcus aureus, and one as a pocket abscess secondary to Staphylococcus epidermidis . Infections were significantly more common when the operator was inexperienced (< or = 100 previous patients), the operation was prolonged, or after a repeat operation for non-infective complications (principally lead displacement) . Infection was not significantly more common in patients identified preoperatively as being at high risk (for example patients with diabetes mellitus, patients receiving long term steroid treatment), although there was a trend in this direction . CONCLUSIONS--Antibiotic prophylaxis significantly reduced the incidence of infective complications requiring a repeat operation after permanent pacemaker implantation . It is suggested that antibiotics should be used routinely.

Eur J Nucl Med, 1994 Oct, 21(10), 1135 - 40
Optimized localization of bacterial infections with technetium-99m labelled human immunoglobulin after protein charge selection; Welling M et al.; To improve the scintigraphic detection of bacterial infections a protein charge-purified fraction of polyclonal human immunoglobulin was applied as a radiopharmaceutical . This purification was achieved by attaching the immunoglobulin to an anion-exchanger column and by obtaining the column-bound fraction with buffer . The binding to bacteria in vitro and the target to non-target ratios of an experimental thigh infection with Staphylococcus aureus or Klebsiella pneumoniae in mice were evaluated to compare the purified and the unpurified immunoglobulin . The percentage of binding to all gram-positive and gram-negative bacteria used in this study was significantly (P < 0.03) higher for the purified than for the unpurified immunoglobulin . For the in vivo study, mice were infected in the thigh muscle with Staph . aureus or K . pneumoniae . After 18 h 0.1 mg of technetium-99m labelled polyclonal immunoglobulin or 99mTc-labelled protein charge-purified polyclonal human immunoglobulin was administered intravenously . At all time intervals the target (infected thighs) to non-target (non-infected thighs) ratios for both infections were significantly higher (P < 0.03) for protein charge-purified polyclonal immunoglobulin than for unpurified polyclonal human immunoglobulin . Already within 1 h the infected tissues could be detected by the purified immunoglobulin . It is concluded that 99mTc-labelled protein charge-purified immunoglobulin localizes both a gram-positive and a gram-negative thigh infection more intensely and faster than 99mTc-labelled unpurified immunoglobulin.

J Clin Microbiol, 1994 Oct, 32(10), 2505 - 9
Quantitative antibiogram typing using inhibition zone diameters compared with ribotyping for epidemiological typing of methicillin-resistant Staphylococcus aureus; Blanc DS et al.; Antibiogram typing of methicillin-resistant Staphylococcus aureus with selected antibiotics was evaluated as a primary epidemiological typing tool and compared with ribotyping . Antibiograms were derived with the Kirby-Bauer disk diffusion method by using erythromycin, clindamycin, cotrimoxazole, gentamicin, and ciprofloxacin . For typing, antibiogram data were analyzed by similarity analysis of disk zone diameters (quantitative antibiogram typing) . One hundred seventy-two isolates were typed . Reproducibility reached 98% for the quantitative antibiogram and 100% for ribotyping . With three selected restriction enzymes (EcoRV, HindIII, and KpnI), 40 epidemiologically unrelated isolates could be classified into 21 ribotypes, whereas quantitative antibiogram typing classified these isolates into 19 groups . To evaluate the discriminatory power of the methods, we calculated an index of discrimination from data obtained with these 40 isolates . This index takes into consideration both the number of types defined by the typing method and their relative frequencies . With both ribotyping and quantitative antibiogram typing, high discrimination indices (0.972 and 0.954, respectively) were obtained . When epidemiological links between patients (ward, period of hospitalization, and contacts between staff and patients) were compared with the results of ribotyping or the quantitative antibiogram typing method, it appeared that both methods were able to discriminate epidemiological clusters, with only a few discrepancies . In conclusion, quantitative antibiogram typing, although not necessarily based on genomic markers, is a simple method which enables a reliable workup of methicillin-resistant S . aureus epidemic when sophisticated molecular typing methods are not available.

J Clin Microbiol, 1994 Oct, 32(10), 2407 - 12
Genetic variation in Staphylococcus aureus coagulase genes: potential and limits for use as epidemiological marker; Schwarzkopf A et al.; To perform coagulase gene typing, the repeated units encoding hypervariable regions of the Staphylococcus aureus coagulase gene were amplified by the PCR technique; this was followed by AluI restriction enzyme digestion and analysis of restriction fragment length polymorphism (RFLP) patterns . In order to assess the discriminatory power of this typing method, 30 epidemiologically unrelated S . aureus strains which differed by their pulsed-field gel electrophoresis patterns were examined . Although 18 of the 30 strains had unique and unshared AluI RFLP patterns, there were only four observed patterns in the remaining 12 strains . This finding indicated that unrelated strains may share identical AluI RFLP patterns . To elucidate the degree of genetic variation in the C-terminus-encoding loci within the coagulase genes, the PCR products of these 12 strains were subjected to Taq polymerase-mediated sequencing . Sequence analysis confirmed the AluI recognition sites in each of the four RFLP groups and demonstrated that AluI appears to yield the highest RFLP in restriction enzyme analysis . By their DNA sequences the majority of strains sharing common AluI groups could be clearly differentiated from each other and revealed between 93.2 and 98.5% homology . When we determined the nucleotide sequences of two strains after six subcultivations no significant alterations were observed . Because the discriminatory power of the current coagulase gene typing method is not great enough to be used as the sole method to type S . aureus, additional techniques are necessary . Sequence analysis of the repeated unit-encoding region for the typing of S . aureus may be potentially useful as an alternative to other current molecular typing techniques.

Jpn J Antibiot, 1994 Oct, 47(10), 1363 - 8
{A study on MRSA infections and replacement of bacteria . Efficacy of vancomycin-ceftazidime combination therapy}; Matsuoka K et al.; We analyzed the results of bacteriological tests on patients with MASA infections admitted to Osaka Prefectural Hospital, for the past 10 years . The conclusions obtained are as follows . 1 . In our hospital, MRSA infections accounted for 50% or more of all Staphylococcus aureus infections in 1983 and 1984 (in the first half of the 1980's), markedly decreased to about 10% after 1988, because of the preventive measures taken against nosocomial infections and initiation of anti-MRSA treatment . During the latter period, use of antibiotics including third generation cephems was not especially restricted . 2 . Glucose non-fermentative Gram-negative rods (GNF-GNR) or yeasts co-isolated with S . aureus were more frequent in MRSA infected patients, than those in MSSA infected patients . 70 to 80% of GNF-GNR were P . aeruginosa . 3 . Sensitivities of MRSA to drugs were studied . VCM was most active, followed by arbekacin and rifampicin . Effectiveness against co-isolated GNF-GNR was high with ofloxacin, netilmicin and ceftazidime (CAZ) . Therefore, is expected that, to prevent the replacement of opportunistic infections, combination therapies using vancomycin and CAZ would be effective.

Clin Infect Dis, 1994 Oct, 19(4), 770 - 3
Treatment with intralesional granulocyte instillations and interferon-gamma for a patient with chronic granulomatous disease and multiple hepatic abscesses; Lekstrom-Himes JA et al.; We present the case of a 16-year-old girl with p22-deficient chronic granulomatous disease in whom multiple hepatic abscesses secondary to Staphylococcus aureus infection developed . Infection persisted despite extensive surgery and aggressive antibiotic therapy . Conventional intravenous granulocyte transfusions were not tolerated because of the development of alloantibodies to HLA . Treatment with interferon-gamma and intralesional granulocyte infusions was associated with dramatic clinical and radiographic improvement . No morbidity was associated with this therapy . To our knowledge, this is the first report of treatment with intralesional granulocyte instillations . Intralesional granulocyte instillation in association with interferon-gamma administration may result in clinical improvement in the conditions of patients with chronic granulomatous disease and hepatic abscesses for whom conventional therapy has failed.

Shock, 1994 Oct, 2(4), 271 - 4
Monophosphoryl lipid A protects against gram-positive sepsis and tumor necrosis factor; Astiz ME et al.; Monophosphoryl lipid A (MPL) is a less toxic derivative of lipid A that enhances survival from endotoxemia . This study examined whether MPL induced resistance to Gram-positive sepsis and cytokines . Mice were administered MPL or saline (phosphate-buffered saline) and challenged 24 h later with live Staphylococcus aureus (SA), staphylococcus enterotoxin B (SEB), toxic shock syndrome toxin (TSST-1), and tumor necrosis factor (TNF) . Survival was determined at 72 h . A separate set of animals was phlebotomized for determination of cytokines . MPL increased survival from S . aureus bacteremia from 20 to 87% (p < .05) . Interleukin-6 (IL-6) and interleukin-1 (IL-1) and TNF were also significantly decreased . SEB and TSST survival were enhanced from 10 to 90% (p < .05) . In SEB-treated animals, TNF and IL-6 levels were significantly decreased . Survival from TNF infusion was increased from 20 to 100% with MPL, however, no significant differences in cytokines were observed . These data suggest that MPL induces resistance to Gram-positive sepsis and cytokine-mediated activity.

Microb Pathog, 1994 Oct, 17(4), 239 - 51
The effect of growth temperature on Staphylococcus aureus binding to type I collagen; Clark BA et al.; Many strains of Staphylococcus aureus produce a collagen-binding surface protein that could enable these strains to colonize tissues such as bone . Previous studies indicated that the expression of the collagen receptor varies with growth conditions . We report here that the growth temperature influences the ability of some S . aureus strains to produce this receptor . S . aureus isolates from human, osteomyelitic bone were grown at 37 degrees C and 42 degrees C and tested for agglutination of collagen-coated latex beads . Binding by 42 degrees C grown cells was significantly reduced in five of the seven isolates studied, including a complete loss of collagen binding in three of these isolates . In an 125I-collagen-binding assay, the binding ability of one of these isolates, strain #16, was 20-fold lower if grown at 42 degrees C . Reduced collagen binding by this isolate could be demonstrated after only two cell divisions at 42 degrees C and the cells regained the ability to bind collagen when shifted back to 37 degrees C . Sodium dodecyl sulfate (SDS)-PAGE confirmed the presence of proteins at 117 kDa in strain #16 and 135 kDa in SMH which were absent following growth at 42 degrees C . Chicken IgG, specific for the 117 kDa protein, was found to react in immunoblot assays with these proteins as well as a protein of 135 kDa extracted from S . aureus Cowan 1 . The antibody did not react with proteins extracted from non-binding strains . Strains #15 and #21, collagen-binders at both 37 degrees C and 42 degrees C, produced immunoreactive proteins at 110 and 135 kDa, respectively, in lysates from cells grown at both temperatures . Antibody against a recombinant form of a previously characterized collagen receptor was used to confirm cross-reactivity with these novel collagen receptors . These data suggest that the ability to produce the collagen receptor is temperature sensitive in some S . aureus strains associated with osteomyelitis . It is proposed that a better understanding of the environmental effects on collagen receptor production could enhance our understanding of staphylococcal infections in bone and joints.

Comp Biochem Physiol B Biochem Mol Biol, 1994 Oct-Nov, 109(2-3), 261 - 71
The primary structure of a basic (pI 9.0) fatty acid-binding protein from liver of Gallus domesticus; Ceciliani F et al.; The complete amino acid sequence of a basic (pI 9.0) fatty acid-binding protein purified from liver of Gallus domesticus was determined by automated Edman degradation of tryptic, CNBr/HFBA and Staphylococcus aureus protease peptides . The protein contains 125 amino acid residues which correspond to a molecular mass of 14094 . The identification of the blocked N-terminus Ac-Ala required digestion of a SV-8 peptide with the acylamino acid-releasing enzyme prior to sequence analysis . Sequence comparison shows that chicken liver basic-FABP has a significant similarity to other proteins belonging to the superfamily of intracellular lipid molecule binding proteins . Moreover, these sequence data confirm that basic-FABP probably binds its substrate in a slightly different way when compared with other FABPs . Basic-FABP was submitted to the EMBL Data Library with an accession number of P80226.

Agents Actions, 1994 Oct, 42(3-4), 135 - 40
Lymphatic mast cell response and effect of compound 48/80 on popliteal lymph node reaction in rats following intracutaneous injection of Staphylococcus aureus; Sudo LS et al.; To investigate the significance of mast cells in the popliteal lymph node during the development of an inflammatory response, rats were inoculated with 12 x 10(7) colony-forming units of Staphylococcus aureus in the hind foot pad . Numerical changes in mast cells were then measured in the corresponding popliteal lymph node . Six days after inoculation, despite the enlargement of the responding lymph node, a marked decrease in granulated mast cell number, relative to the contralateral node, was observed in the cortical and medullary compartments . Popliteal lymph nodes from rats treated with compound 48/80 and then inoculated with S . aureus showed a higher cortical and medullary hypertrophic response and a significant increase in degranulated/weakly basophilic mast cell number in the lymph node tissue . The findings suggest that (1) Staphylococcus aureus induces a reduction in granulated mast cell number in the cortical and medullary compartments of regional lymph nodes; (2) pretreatment with compound 48/80 appears to contribute to the lymphoid cell proliferation and the hypertrophic response of lymph nodes induced by S . aureus; and (3) granulated mast cells have a regulatory role on lymphoid cell proliferation.

J Med Chem, 1994 Sep 30, 37(20), 3205 - 11
"Glycylcyclines" . 3 . 9-Aminodoxycyclinecarboxamides; Barden TC et al.; A series of 9-(acylamino)doxycycline derivatives has been prepared . These analogs exhibit good activity against both tetracycline sensitive and tetracycline resistant Gram-positive (Staphylococcus aureus) and Gram-negative (Escherichia coli) bacteria that are encoded with the efflux and ribosomal resistance gene factors . N,N-Dialkylglycylamido derivatives possessed the highest activity . Replacement of glycine moiety with other amino acids did not further enhance the activity.

J Biol Chem, 1994 Sep 23, 269(38), 23530 - 7
Identification of a distinct pool of sphingomyelin involved in the sphingomyelin cycle; Linardic CM et al.; Sphingomyelin (SM) is a membrane phosphosphingolipid that has recently been identified as a key component of the SM cycle . In this signal transduction pathway, extracellular inducers such as tumor necrosis factor alpha cause hydrolysis of membrane SM, resulting in the generation of the lipid second messenger ceramide . Only 10-20% of cellular SM appears to be involved in the SM cycle, raising the possibility of the existence of a unique "signaling" pool of SM . The existence and subcellular location of such a pool were investigated . Using bacterial sphingomyelinase from Staphylococcus aureus (bSMase), we first characterized two pools of SM, identified as an outer leaflet bSMase-sensitive pool and a distinct bSMase-resistant pool . These pools were further characterized by their differential solubility in Triton X-100 and by their kinetics of labeling . The signaling pool of SM was distinguished by the following: 1) resistance to bSMase, 2) solubility in Triton X-100, and 3) delayed labeling kinetics . In subfractionation studies, the signaling pool of SM co-fractionated with the plasma membrane . Since the SM cycle involves a cytosolic sphingomyelinase and the intracellular release of choline phosphate, this pool of SM appears to localize to the inner leaflet of the plasma membrane (or to a closely related compartment) . These results identify a unique signaling pool of SM that undergoes significant hydrolysis (20-40%) in response to inducers of the SM cycle.

Ugeskr Laeger, 1994 Sep 19, 156(38), 5497 - 502
{Nosocomial pneumonias . The effect of selective intestinal and ventricular decontamination in respirator-treated intensive care patients}; Engelsen J et al.; Nosocomial infections, especially pneumonias, are a common problem in Intensive Care Units (ICU) and are a major cause of morbidity and mortality . During the past ten years a new regime, selective decontamination of the digestive tract (SDD) has been introduced to prevent these infections which are often preceded by colonization with aerobic potentially pathogenic microorganisms . Topical antibiotics are applied both orally and by nasogastric tube to prevent secondary endogeneous infections with Gram-negative enteric rods, Staphylococcus aureus and yeasts, in combination with parenteral antibiotics for the treatment of early primarily exogeneous and endogeneous infections . The effect of SDD on morbidity and mortality has been studied in many papers which are very conflicting and their results are difficult to interpret . This article reviews 11 published randomized trials of SDD in ICU among intubated and artificially ventilated adult patients aged more than 15 years . Of the eleven available controlled studies, eight showed a significant reduction of nosocomial pneumonias among patients who received selective decontamination . Only two studies demonstrated a reduction in infection related mortality rates.

FEMS Microbiol Lett, 1994 Sep 15, 122(1-2), 187 - 93
Simultaneous measurement by flow cytometry of phagocytosis and metabolic burst induced in phagocytic cells in whole blood by Borrelia burgdorferi; Cinco M et al.; This paper describes the interactions between a strain of Borrelia burgdorferi and phagocytic cells, measured in whole blood, by a two-color flow cytometric method, which allowed the simultaneous quantification of both the phagocytosis rate and the oxidative burst activation . The data obtained indicated that: a) phagocytosis and metabolic activation increased as a function of spirochete concentration; b) the number of ingesting cells peaked within 10 min but activation followed later, and did not involve all the phagocytosing cells; c) opsonization of borreliae with a patient's serum enhanced the two cellular activities, mostly phagocytosis . The intensity of such functions was lower than those found for Staphylococcus aureus . The flow cytometric assay of phagocytes interactions with Borrelia burgdorferi assessed in whole blood represents an experimental approach which simulates the physiological conditions in nature.

Proc Natl Acad Sci U S A, 1994 Sep 13, 91(19), 9057 - 61
Binding of soluble natural ligands to a soluble human T-cell receptor fragment produced in Escherichia coli; Hilyard KL et al.; An Escherichia coli expression system has been developed to produce milligram quantities of the variable domains of a human T-cell receptor from a cytotoxic T cell that recognizes the HLA-A2-influenza matrix peptide complex as a single polypeptide chain . The recombinant protein was purified by metal-chelate chromatography and then refolded in a redox buffer system . The refolded protein was shown to directly bind both Staphylococcus aureus enterotoxin B and the major histocompatibility complex protein-peptide complex using a BIAcore biosensor . Thus this preparation of a single-chain, variable-domain, T-cell receptor fragment can bind both of its natural ligands and some of it is therefore a functional fragment of the receptor molecule.

Zentralbl Veterinarmed A, 1994 Sep, 41(7), 558 - 67
Phagocytic activity of polymorphonuclear leukocytes lavaged from the lungs of horses with clinically diagnosed chronic pulmonary disease; Klucinski W et al.; The aim of this study was to compare phagocytic activity of polymorphonuclear cells (PMNs) from the bronchoalveolar lavage of clinically healthy horses and those with severe chronic bronchiolitis . Research was carried out on 28 horses . Chronic inflammation of the lower airways was diagnosed in nine horses . Cells from the respiratory tract were lavaged according to accepted methods . For comparison, PMNs were isolated from peripheral blood of all investigated horses . The phagocytic activity of PMNs was determined in relation to two standard strains of Staphylococcus aureus, Staph, aureus Smith which was phagocytized after previous opsonization, and Staph, aureus 305, phagocytized without opsonization . From the investigations, it is shown that the PMNs present in the terminal airways of horses with severe chronic bronchiolitis are characterized by decreased phagocytic activity in relation to opsonized Staphylococcus aureus Smith and increased activity in relation to non-opsonized Staphylococcus aureus 305, as compared to the PMNs lavaged from the terminal airways of clinically healthy horses . No changes in the phagocytic activity of the peripheral blood PMNs were observed between clinically diseased horses and healthy horses.

Biochem J, 1994 Sep 1, 302 ( Pt 2), 503 - 9
Limited proteolysis of phospholipase C-gamma 1 indicates stable association of X and Y domains with enhanced catalytic activity; Fernald AW et al.; Phospholipase C-gamma 1 (PLC-gamma 1) was treated with Staphylococcus aureus V8 protease (V8) and the digestion products were analysed with site-specific antibodies . V8 treatment generated three immunodetectable PLC-gamma 1 fragments of 120, 97, and 39 kDa . The 39 kDa fragment is derived from the C-terminus of PLC-gamma 1 and includes the conserved Y domain present in all PLC isoenzymes . The 120 and 97 kDa fragments are derived from the N-terminus of PLC-gamma 1, possess the conserved X domain common to all PLC isoenzymes, and the src-homology domains unique to PLC-gamma 1 and -gamma 2 . It is likely that the 97 kDa fragment is a V8 product of the 120 kDa fragment . As the C-terminal 39 kDa fragment, and either of the N-terminal 120 or 97 kDa fragments, were precipitable with antibody specific to a sequence present in only the 39 kDa fragment, the data indicate co-precipitation of separate polypeptide chains that remain associated after V8 proteolysis . Importantly, V8 treatment increased the activity of PLC-gamma 1 and did not alter the calcium requirement . The influence of other modulators of PLC-gamma 1 activity, however, was lost following V8 treatment . These results suggest the stable association of the X and Y domains within PLC-gamma 1, and demonstrate that proteolysis in the region of PLC-gamma 1 that is subject to tyrosine phosphorylation can enhance catalytic activity.

Am J Physiol, 1994 Sep, 267(3 Pt 2), H952 - 61
Modulation of uterine resistance artery lumen diameter by calcium and G protein activation during pregnancy; D'Angelo G et al.; The purpose of this study was to determine whether the increased sensitivity of uterine resistance arteries from late pregnant (LP) rats to alpha-adrenergic stimulation is due to an alteration in the fundamental relationship between cytosolic calcium (Ca2+) and arterial lumen diameter . Uterine arcuate arteries were permeabilized with Staphylococcus aureus alpha-toxin under optimal conditions and constricted to varying degrees with discrete Ca2+ concentrations at a distending pressure of 50 mmHg . Arterial segments from nonpregnant (NP) and LP rats exhibited similar Ca2+/lumen diameter characteristics . Ca2+ (0.1 microM) produced appreciable constriction, and lumen diameter decreased steeply between 0.175 and 0.25 microM Ca2+; maximal responses were attained with 0.5 microM Ca2+ . Activation of guanine nucleotide binding proteins (G proteins) with guanosine 5'-triphosphate (GTP; 1-100 microM), as reportedly occurs during alpha-adrenergic stimulation, potentiated the Ca(2+)-induced constriction by 121 and 79% in arteries from LP and NP rats, respectively . No significant differences between the two animal groups were noted . Guanosine 5'-O-(gamma-thiotriphosphate) (GTP gamma S; 0.1-10 microM), a nonhydrolyzable analogue of GTP, effected a larger potentiating effect over that maximal response caused by GTP in arteries from NP rats . Ca(2+)- and Ca2+/GTP-induced constrictions were more potently reversed by guanosine 5'-O-(beta-thiodiphosphate) (GDP beta S)., a competitive inhibitor of GTP, in arteries from NP rats . These data suggest that pregnancy-induced increases in sensitivity to alpha-adrenergic stimulation may be related to altered G protein cycling rates, such that G proteins in smooth muscle cells in arcuate arteries from NP rats are more susceptible to deactivation . Alternatively, consistent with the model of G protein-mediated inhibition of myosin light chain phosphatase, myosin light chain phosphatase activity may be enhanced in uterine vascular smooth muscle from NP rats relative to that from LP rats.

J Bone Joint Surg Br, 1994 Sep, 76(5), 717 - 20
Influence of metal implants on infection . An experimental study in rabbits; Cordero J et al.; We implanted cylinders of cobalt-chrome or titanium, with smooth or porous surfaces, into rabbit bones which had been inoculated with suspensions of Staphylococcus aureus in various doses . The bacterial concentration required to produce infection of porous-coated titanium implants was 2.5 times smaller than that necessary to infect implants with polished surfaces . Porous-coated cobalt-chromium implants required bacterial concentrations that were 40 times smaller than those needed to infect implants with polished surfaces, and 15 times smaller than those required to infect porous-coated titanium implants . The other advantages and disadvantages of the various implants, such as improved osseointegration, larger ion-release surfaces, surface wear and relative stiffness, must be weighed against the higher infection rates in the porous-coated implants, and particularly in the cobalt-chromium implants.

Int J Cancer, 1994 Sep 1, 58(5), 678 - 85
EBV infection of B-CLL cells in vitro potentiates their allostimulatory capacity if accompanied by acquisition of the activated phenotype; Avila-Carino J et al.; Epstein-Barr virus (EBV)-carrying immortalized lymphoblastoid cell lines (LCLs) stimulate autologous T lymphocytes in vitro . This T-cell response is independent of the EBV-specific cellular memory because it also occurs in experiments with cells of seronegative individuals . The question can be posed whether the T-cell-stimulatory potential of the LCL is coupled to its immortalized state . B-CLL cells were exploited to study this question because the majority of clones, represented by different patients, can be infected with EBV but they rarely become immortalized . We have investigated the phenotypic changes and the T-cell-stimulatory capacity of EBV-infected B-CLL cells . One aliquot of CLL cells was infected with EBV, another was activated with a mixture of Staphylococcus aureus (SAC), IL-2 and the supernatant from the T-cell hybridoma MP6 (activation mixture, AcMx) and the third aliquot received both treatments . In accordance with the individual features of the clonal populations represented by each patient, the immunophenotypic changes imposed by these treatments differed . With the samples of 3 patients the allo-stimulatory potential showed the following ranking order: EBV and AcMx-treated cells > AcMx-treated > EBV-infected . An analysis of several activation-related surface markers and adhesion molecules on the cells did not reveal any association between their expression and the EBV-imposed potentiation of allostimulatory capacity . These results may be extrapolated to EBV-genome-carrying normal B cells, suggesting that they can persist in vivo only as long as they have the resting phenotype . Once they are activated, these cells may be recognized and eliminated by T lymphocytes.

J Bacteriol, 1994 Sep, 176(17), 5534 - 6
Coagulase expression in Staphylococcus aureus is positively and negatively modulated by an agr-dependent mechanism; Lebeau C et al.; Expression of staphylocoagulase by agr+ Staphylococcus aureus depends on the growth phase, being maximal during exponential growth and decreasing sharply postexponentially, while an agr-deleted strain continuously expresses an intermediate level of coagulase . Therefore, coagulase expression appears to be both positively and negatively modulated by an agr-dependent mechanism.

Cell Immunol, 1994 Sep, 157(2), 478 - 88
The interplay of interleukin-10 (IL-10) and interleukin-2 (IL-2) in humoral immune responses: IL-10 synergizes with IL-2 to enhance responses of human B lymphocytes in a mechanism which is different from upregulation of CD25 expression; Itoh K et al.; The role of the interplay of interleukin-2 (IL-2) and interleukin-10 (IL-10) in responses of human peripheral blood B cells stimulated by ligation of antigen receptors was examined in detail . Highly purified peripheral blood B cells from normal human individuals were cultured with Staphylococcus aureus Cowan I (SA) in the presence or absence of IL-10 and IL-2 . Although IL-10 alone could modestly induce Ig production by SA-activated B cells, IL-10 markedly enhanced the Ig production in synergism with IL-2 . This synergistic effect between IL-2 and IL-10 was completely abrogated by addition of either anti-CD25 mAb or anti-IL-10 mAb, indicating signals through high-affinity IL-2 receptors are involved in the synergism . In fact, IL-10 upregulated the expression of alpha-chain of IL-2 receptors (CD25) on B cells stimulated by SA alone or SA + IL-2 for 72 hr . However, such upregulation of CD25 expression did not result in significant enhancement of subsequent responses of the activated B cells to IL-2 . Of note, during the initial activation phase of B cells, IL-2 was much more effective than IL-10 in enhancing the subsequent responses to either IL-2 or IL-10 . By contrast, IL-10 was much more effective than IL-2 in supporting the maturation of B cells following the initial activation . Finally, the synergy between IL-2 and IL-10 was not observed in the initial activation phase, but in the subsequent maturation phase of activated B cells . These results indicate that IL-10 and IL-2 synergistically enhance Ig production of SA-activated B cells in a mechanism which is different from the upregulation of IL-2 receptors . Moreover, the data emphasize the importance of the interplay of IL-2 and IL-10 in determining the outcome of humoral immune responses.

Cell Immunol, 1994 Sep, 157(2), 381 - 92
Enhancing effects of interleukin 2-treated peripheral blood mononuclear cells on subsequent B cell differentiation; Stohl W et al.; Despite an extensive literature dealing with IL2-induced cytolytic activity, noncytotoxicity-related effects of IL2 on peripheral blood mononuclear cells (PBMC) or T cell function have received less attention . We have focused on the effects of irradiated, IL2-activated PBMC (PBMC*rIL2) on anti-CD3- and formalin-fixed heat-killed Staphylococcus aureus-induced polyclonal B cell differentiation in secondary cultures . PBMC*rIL2 act directly on B cells and cross major histocompatibility complex barriers to augment polyclonal B cell differentiation as measured by plaque-forming cell (PFC) generation . These effects are preferentially mediated by T (both CD4+ and CD8+) cells, and physical contact between effector PBMC*rIL2 and target B cells is not absolutely required for enhanced PFC generation . PBMC*rIL2 must be present for the initial 24 hr of the secondary cultures, indicating that some soluble B cell differentiation factor rapidly released by PBMC*rIL2 mediates the PFC-enhancing effect . Of IL2, IL4, IL5, IL6, IL10, IFN-gamma, and TNF-alpha, only IFN-gamma mRNA is appreciably and reproducibly increased in irradiated, IL2-activated T cells (T cells*rIL2) . Nevertheless, exogenous rIFN-gamma cannot mimic and anti-IFN antibodies cannot block the PFC-enhancing effects of T cells*rIL2, indicating that some unidentified soluble factor(s) apart from or in addition to IFN-gamma is involved . IL2-induced effects on T cell noncytolytic function may help explain certain observed immune anomalies in IL2-treated patients, and a better understanding of the IL2-induced effects on T cell noncytolytic function may have ramifications for autoimmune diseases such as SLE.

Infect Immun, 1994 Sep, 62(9), 3780 - 5
A new type of staphylococcal exfoliative toxin from a Staphylococcus aureus strain isolated from a horse with phlegmon; Sato H et al.; A new type of staphylococcal exfoliative toxin (sET) was isolated from the culture filtrate of a Staphylococcus aureus strain isolated from a horse with skin infection including phlegmon . The new sET was purified by precipitation with 80% saturated ammonium sulfate, column chromatography on DEAE-cellulofine A-500, gel filtration on a Sephadex G-75 column, and polyacrylamide gel electrophoresis (7.5% polyacrylamide) . The new sET elicited general exfoliation of the epidermis with the so-called Nikolsky sign when inoculated into both 3-day-old mice and 1-day-old chicks, whereas sETA and sETB from human strains of S . aureus caused exfoliation in a 3-day-old mouse alone and shET from a porcine strain of Staphylococcus hyicus caused exfoliation in 1-day-old chicks alone . Intraepidermal splitting was observed at the granular layer of the epidermis of mice inoculated with the new sET as well as those inoculated with sETA . Exfoliation at the germinative layer of the epidermis was also observed in the chicks inoculated with the new sET as well as those inoculated with shET . The new sET was serologically different from sETA, sETB, and shET and showed the same molecular weight on sodium dodecyl sulfate-polyacrylamide gel electrophoresis . It was thermolabile and lost its toxicity after being heated at 60 degrees C for 15 min . We propose that the new sET be designated as sETC.

Clin Immunol Immunopathol, 1994 Sep, 72(3), 394 - 401
Molecular basis for the interaction between human IgM and staphylococcal protein A; Hakoda M et al.; To examine the relationship between VH gene usage and reactivity of immunoglobulins, we cloned B cells from peripheral blood from adults and from human neonatal cord blood by EBV transformation . Nearly one-third of the B cell clones from both sources produced IgM reactive with staphylococcal protein A (SPA) . None of such IgM reacted with other antigens, except for the crude extract of Staphylococcus aureus . All of 22 B cell clones producing IgM reactive with SPA expressed VH3 genes, while none of the control 15 clones secreting IgM nonreactive with SPA expressed VH3 . The IgM proteins reactive with SPA could be clearly divided into two subjects based on the differential binding avidity to solid-phase SPA . Both kappa and lambda light chains were used in each subset of SPA-reactive IgM . Sequence analysis of the PCR products from seven VH3-IgM clones revealed that the VH3 genes were used in nearly germline configuration . The D and J gene usage was diverse . Comparison of amino acid sequences between antibodies with high and low avidity to SPA suggests that the differential avidity is related to amino acid sequence differences in the complementarity determining region 2 and framework region 3 . The high frequencies of B cells committed to the production of SPA-reactive IgM in normal blood and the restricted use of VH3 heavy chain genes in nearly germline configuration in these cells support the notion that SPA behaves like a superantigen toward human B cells.

J Am Osteopath Assoc, 1994 Sep, 94(9), 741 - 3
Automotive airbag-induced second-degree chemical burn resulting in Staphylococcus aureus infection; Polk JD et al.; A young woman did not seek emergency treatment after a minor automobile collision as she thought that she had been spared serious injury by the inflation of the driver's-side airbag . She had a benign-looking erythema on her neck which, over the next several days, became a second-degree chemical burn infected with Staphylococcus aureus . The burn and subsequent infection took several weeks to heal and the patient had to endure a prolonged course of antibiotics, nonsteroidal antiinflammatory drugs, and continued irrigation . This case exemplifies why alkali chemical burns from an automotive airbag should be treated aggressively, despite their benign appearance, as they may take several days to evolve . Physicians should be warned that careful follow-up examination of patients seen in the hospital emergency department or in the physician's office is necessary to abate any hidden sequelae . Of course, the opportunity to decrease morbidity is lost if the patient does not seek emergency treatment.

Minerva Chir, 1994 Sep, 49(9), 807 - 11
{Incidence and treatment of vascular prosthesis infection in the groin . Personal experience}; Bonelli U et al.; The incidence of prosthesis infection in the groin ranges between 1 and 3%, with high morbidity and mortality rates . Contamination, irrespective of whether it is endogenous or exogenous, may occur during surgery of afterwards due to a secondary bacterial load which is often associated with a traumatic surgical technique for tissues, often in emergency surgery or re-operations . MATERIALS AND METHODS . Over the past 5 years a total of 410 prosthetic alloplastic grafts have been made by our Division of Vascular Surgery with at least one anastomosis at an inguinal level . The materials used for prosthesis were dacron and PTFE . Thirty-one infections were observed in the groin of which 11 involved the prosthesis and 20 were apparently restricted to the superficial and deep layer of the dermis . A cell culture test and antibiogram was always performed . Surgical or medical therapy (loco-regional + general) was used according to individual cases . RESULTS . With regard to the 11 patients with manifest infections of the prosthesis, only trans-obturator bypass surgery gave positive long-term results, whereas other operations (femoro-femoral bypass, axillo-popliteal bypass) were not satisfactory . Of the other 20 patients treated using loco-regional and general medical therapy, 9 developed pseudo aneurysm over time for which it was then necessary to resort to surgery . DISCUSSION AND CONCLUSION . Currently there is no universally accepted treatment protocol for the management of prosthesis infection . However, it is generally considered useful to isolate any pathological agent (using routine biopsies of arterial wall or thrombo-endoluminal fragments during the primary operation) before infection can develop so as to be able to commence specific antibiotic therapy . In this context the authors underline the more devastating action of some bacteria (pseudomonas, Staphylococcus aureus) in comparison to others (Staphylococcus epidermidis) . Turning to therapeutic measures, when there are anatomical grounds and with the greatest possible respect for periarterial tissues, it is considered that trans-obturator bypass is preferable in the presence of manifest infection of the prosthesis, whereas medical therapy can be used (loco-regional + general) if infection is limited to the surface layers of the inguinal area . Possible complications over time may be corrected using surgery and in situ reconstruction if infection is no longer present.

Free Radic Res, 1994 Sep, 21(3), 147 - 67
The bactericidal action of peroxides; an E.P.R . spin-trapping study; Clapp PA et al.; E.P.R . spin trapping has been employed to study radical production during the bactericidal action of three peroxide compounds (peracetic acid, 4-percarboxy-N-isobutyltrimellitimide and magnesium monoperoxyphthalate) upon both Gram negative (Escherichia Coli) and Gram positive (Staphylococcus Aureus) bacteria . Use of the spin trap 5,5-dimethyl-1-pyrroline N-oxide (DMPO) has allowed direct detection of both carbon-centred and hydroxyl radicals, which are produced at varying rates for the different bacteria/peracid systems studied . The inhibition of bactericidal action, by DMPO and two antioxidants, Vitamin C and Trolox C, indicates that radicals are the lethal species and evidence is presented which suggests that radical production is internal to the bacterial cell . Hydroxyl radicals are believed to be the lethal species . The effect of added iron chelators and haem protein inhibitors indicates that iron species and haem proteins in particular are involved . A marked variation is found in observed hydroxyl-radical adduct signals with both the nature and concentration of peracid . A strong inverse correlation is found between the concentration of the observed radical adduct signal and the relative strength of the peroxide as a bactericide; use of a stable nitroxide as a radical scavenger confirms that strong bactericides produce radicals at a much faster rate than weak bactericides . Plots of radical generation versus time are correlated with % bacterial kill, offering further evidence that hydroxyl radicals are the lethal species.

Rinsho Byori, 1994 Sep, 42(9), 953 - 60
{Multi-center evaluation to discriminate between the strains of methicillin-resistant Staphylococcus aureus (MRSA) and those susceptible (MSSA) by Showa oxacillin and methicillin disk susceptibility tests}; Takahashi A et al.; The Showa disk susceptibility test using two penicillinase-resistant penicillins, oxacillin and methicillin, was evaluated to discriminate between the strains of Staphylococcus aureus resistant to methicillin (MRSA) and those susceptible (MSSA) in the multi-center trials . The study included 651 clinical isolates of S . aureus, comprising of 329 MRSA and 322 MSSA isolates . The inhibitory zone diameters by Showa disks to oxacillin and methicillin highly correlated with minimum inhibitory concentrations (MICs) determined by standard agar dilutions with 0.961 and 0.930 correlation coefficients, respectively . Of 651 duplicate MIC determinations, 79.9% (oxacillin) and 80.3% (methicillin) were within +/- 1 log2 dilutions with each other . When Showa oxacillin and methicillin disks were incubated at 35 degrees C, sensitivity and specificity of oxacillin to detect MRSA were 95.4% and 98.1%, and those of methicillin were 94.8% and 95.2% . When tested on agar plates supplemented with 5% NaCl, sensitivity and specificity markedly improved to > 97% . Also, when incubated at 30 degrees C, sensitivity and specificity became to nearly 100% . Of 329 MRSA isolates, the interpretive criteria combined with incubation at 30 degrees C and testing onto 5% NaCl supplemented agar plates could correctly identify 324 (98.5%) and 329 (100%) isolates, respectively . In conclusion, when the Showa oxacillin and methicillin disk susceptibility tests were employed exactly according to the manufacturer's instruction, the test performances to detect MRSA were enough reliable to screen MRSA isolates in clinical microbiology laboratories.

J Dermatol, 1994 Sep, 21(9), 655 - 9
Factors responsible for death in patients with pemphigus; Kanwar AJ et al.; The records of 10 patients who died with pemphigus have been examined for factors affecting and contributing to death . Early initiation of therapy, the age at onset of the disease, and the mode of administration of corticosteroids, conventional or in pulse form, did not affect the survival . The cutaneous involvement was extensive in all 10 patients; it ranged between 30-80% . Septicemia was the commonest event preceding death; in 4 cases, it was due to Staphylococcus aureus.

Chemotherapy, 1994 Sep-Oct, 40(5), 304 - 10
Effects of subinhibitory concentrations of seven macrolides and four fluoroquinolones on adhesion of Staphylococcus aureus to human mucosal cells; Braga PC; Reaction of bacterial adhesins with complementary receptors on the surfaces of mucosal respiratory, gastrointestinal and genitourinary cells leads to bacterial adhesion to the human body . This is the first step in the sequence of events leading to infection . It has been observed that subinhibitory concentrations (sub-MICs) of some antibiotics are able to reduce bacterial adhesion . The pharmacokinetic curves of antibiotics show that these sub-MICs are present in tissues during a typical course of therapy . This study investigated the ability of sub-MICs of seven macrolides and four fluoroquinolones to reduce adhesiveness of Staphylococcus aureus to human buccal cells . The literature generally reports data for only one antibiotic at a time . To obtain an overall view of the findings of eleven antibiotics together, the data have been normalized according to the molecular weight of each antibiotic and the dose-effect curves determined.

Cardiovasc Res, 1994 Sep, 28(9), 1378 - 84
Effect of altered bathing pH on calcium activated force in alpha toxin permeabilised rat portal vein and human umbilical artery; Crichton CA et al.; OBJECTIVE: The aim was to investigate the effect of altered bathing pH on calcium activated force generated by the contractile proteins in an alpha toxin permeabilised phasic (rat portal vein) and tonic (human umbilical artery) smooth muscle . METHODS: Strips of the two muscles were permeabilised using crude alpha toxin from Staphylococcus aureus (2 mg.ml-1) . The relationship between {Ca2+} and tension was then examined at different bathing pH values . {Ca2+} was monitored using indo-1 fluorescence . RESULTS: GTP-gamma-S (100 microM) potentiated maximum calcium activated force in rat portal vein on average to 146%, but had no significant effect on human umbilical artery, confirming the difference in contractile behaviour between the two muscles . Lowering bathing pH from 7.2 to 6.7 depressed submaximal calcium activated force and increased maximum calcium activated force in rat portal vein . Raising bathing pH from 7.2 to 7.7 depressed both submaximal and maximum calcium activated force in rat portal vein . Altered bathing pH had no significant effect on either maximum or submaximal calcium activated force in human umbilical artery . The calcium sensitivity of tension production was not significantly effected by acidic pH in either preparation . However, alkaline pH caused a similar fall in the calcium sensitivity in both preparations . CONCLUSIONS: Tension generated from the contractile proteins of a phasic smooth muscle (rat portal vein) are more sensitive to altered bathing pH than those of a tonic smooth muscle (human umbilical artery).

Rev Esp Fisiol, 1994 Sep, 50(3), 153 - 8
Effect of cefepime (BMY-28142) on the hemolytic and bactericidal activity of serum in vivo and in vitro; Pedrera MI et al.; The present study was undertaken to determine whether cefepime (BMY-28142) could influence the hemolytic and bactericidal activity of serum both in vivo and in vitro . For the in vivo studies, mice (Swiss aged 15 +/- 2 weeks) were studied before and 1, 2, 4, 8, and 12 h after an intramuscular injection of 200 mg/kg of cefepime . Likewise, another group of mice were injected for 7 days with a daily injection of the same dose as before, and sacrificed one hour after the last injection . For the in vitro studies, samples were incubated in the presence and absence of cefepime (1/2, 1/4 and 1/8 MIC against Staphylococcus aureus) (2.5, 1.25 and 0.625 mg/l respectively) . The results indicate that cefepime at sublethal concentrations increases the bactericidal activity of serum both in vivo and in vitro against S . aureus, but does not significantly modify the hemolytic activity of the complement.

J Chemother, 1994 Sep, 6 Suppl 4, 33 - 7; discussion 39-40
Healthcare workers and the incidence of nosocomial infection: can treatment of one influence the other?--a brief review; Wenzel RP; Nasal carriage by health care workers represents an important hospital reservoir of Staphylococcus aureus . Approximately 25% of all hospital-based healthcare workers are stable nasal carriers . Several studies in the US and UK have shown that following treatment of this group with a single 5-day course of intranasal mupirocin, nasal carriage was usually eradicated within 24 hours, and after 12 weeks was only present in 25% of participants . Long-term follow-up in one institution after 52 weeks showed that there were significantly fewer carriers in the mupirocin group than in the group receiving identical placebo . In the same study, between 30% and 50% of those hospital workers who carried S . aureus in their nose, before the start of therapy, were also hand carriers . After treatment, a dramatic reduction in hand carriage of S . aureus was noted, in contrast to no change in the placebo group . After 6 months, the level of hand carriage was still statistically lower in the mupirocin group than in those given placebo . The association between nasal carriage and hand carriage makes it important that health care workers decontaminate their hands effectively between patients . Current evidence suggests, however, that compliance with such control measures is low . Other studies examining the role of S . aureus nasal carriage in the development of post-operative wound infection, have shown that almost half of those isolates recovered from the wound site were present in the nose of the patient pre-operatively . Due to its ability to eliminate nasal carriage of S . aureus, current studies are investigating whether intranasal mupirocin can prevent post-operative wound infections in patients undergoing surgery.

Thromb Haemost, 1994 Sep, 72(3), 403 - 7
Monoclonal purified F VIII for continuous infusion: stability, microbiological safety and clinical experience; Schulman S et al.; Replacement therapy for patients with hemophilia A postoperatively or for major hemorrhage, administered as a continuous infusion, is efficient and reduces the requirement for factor VIII (F VIII) . The convenience of the method is increased by using a minipump and not diluting the concentrate further after reconstitution . A monoclonally purified F VIII concentrate (Monoclate-P), was evaluated for its stability after reconstitution in different infusion systems, for its microbiological safety as well as clinical safety and efficacy in continuous infusion . The F VIII activity was unaffected by 2 of the 3 infusion systems at room temperature during 15 days, whereas in the third (CADD-1) it decreased below 80% of initial value after 3-7 days . Addition of heparin (1 U/ml) or low molecular weight heparin (1 anti-Xa U/ml), which are used to prevent thrombophlebitis at the site of infusion, did not affect the stability . Nine out of 9 samples taken from the infusion systems after 3 days and again after 7 days were sterile . After inoculation with Staphylococcus aureus or Escherichia coli the bacterial growth in samples of the reconstituted concentrate was not different from that in lidocaine in saline or heparin in saline . F VIII was given in continuous infusion with a minipump (Infu-Med) to 12 patients undergoing major surgery and 8 patients with major hemorrhage for a total of 157 days . A progressive decrease of the clearance was seen during the first 5 days of infusion from 3.0 to 1.7 ml/kg/h . Hemostasis was effectively achieved, and no infectious complications were registered.(ABSTRACT TRUNCATED AT 250 WORDS)

Hemoglobin, 1994 Sep, 18(4-5), 275 - 84
The use of Staphylococcus V8 protease in the structural determination of human hemoglobin variants: HB Valparaiso {alpha 88(F9)Ala-->Gly} example; Wajcman H et al.; In human hemoglobin, enzymic cleavage with the protease from Staphylococcus aureus strain V8 (protease V8) is a convenient method for the study of variants that involve large peptides such as alpha T-9 and alpha T-12b . We report here on Hb Valparaiso {alpha 88(F9)Ala-->Gly}, a new neutral variant with a slight increase in oxygen affinity, that was identified by use of this strategy.

Infection, 1994 Sep-Oct, 22(5), 338 - 42
Enhanced killing of methicillin-resistant Staphylococcus aureus in human macrophages by liposome-entrapped vancomycin and teicoplanin; Onyeji CO et al.; The antibacterial effects of liposomal vancomycin and teicoplanin against intracellular methicillin-resistant Staphylococcus aureus (MRSA) were evaluated using a macrophage infection model . Human blood-derived monocytes were cultured for 7 days to obtain adherent macrophages . Uptake of each drug by macrophages was markedly enhanced by liposomal encapsulation . Following phagocytosis and removal of residual extracellular MRSA, the infected macrophages were exposed to clinically achievable concentrations of teicoplanin and vancomycin . The free (untrapped) and liposome-entrapped forms of each drug were used at the same concentration . The number of intracellular surviving bacteria was determined by colony counts after lysis of the macrophages at different time intervals following drug treatment . Intracellular antimicrobial effect of each drug was significantly (p < 0.001) increased by entrapment in liposomes . Also, the efficacies of the free and liposomal forms of both drugs were correspondingly comparable (p > 0.05) . It is, therefore, concluded that liposomal encapsulation of vancomycin and teicoplanin results in an increased availability of the antibiotics for efficient elimination of intracellular MRSA infection.

Eur J Clin Microbiol Infect Dis, 1994 Sep, 13(9), 741 - 6
In vitro development and stability of tolerance to cloxacillin and vancomycin in Staphylococcus aureus; Voorn GP et al.; The stability of tolerance of Staphylococcus aureus during subculturing at 37 degrees C and development of this property after repeated exposure to cloxacillin or vancomycin were investigated in vitro . Four of five tolerant strains lost this property during repeated subculturing at 37 degrees C for 50 days . Conversely, tolerance emerged in two of four nontolerant strains after repeated cycles of exposure to 25 micrograms of cloxacillin/ml or 10 micrograms of vancomycin/ml alternating with growth in antibiotic-free medium . Previous in vivo exposure to cloxacillin did not enhance the development of tolerance in vitro . MICs of both cloxacillin and vancomycin did not change significantly during this procedure . Whether the conversion of nontolerant strains to the tolerant state can also occur during antibiotic exposure in treatment of patients remains to be determined.

J Invest Surg, 1994 Sep-Oct, 7(5), 453 - 65
A new model for posttraumatic osteomyelitis in rabbits; Eerenberg JP et al.; A new animal model for posttraumatic osteomyelitis was designed . This model mimics the pathogenesis of the human disease more accurately than models presently available . Femora of New Zealand white rabbits were exposed at the greater trochanter and a stainless steel rod was inserted into the marrow cavity . A Staphylococcus aureus suspension was placed in and around a bone defect, which was drilled midshaft . The disease was evaluated by clinical observation and roentgenographic, hematologic, bacteriologic, and histologic parameters . Osteomyelitis developed in all 24 infected rabbits . None of the five rabbits receiving only an intramedullary rod developed an osteomyelitis . This model proves that an experimental posttraumatic osteomyelitis associated with a foreign body can be reliably induced, even when no infection-promoting chemical agents, small inoculum of bacteria, or minimal bone trauma is present.

J Craniofac Surg, 1994 Sep, 5(4), 247 - 52; discussion 253
Bacterial exposure required to induce rabbit calvarial bone graft infection by superficial contamination; Lalikos JF et al.; Little is known about the minimum bacterial dose required to induce infections by superficial contamination alone (i.e., not direct inoculation) . This study quantified superficial rabbit calvarial bone graft contamination by exposing parietal bone grafts to Staphylococcus aureus . Two 5 x 5-mm full-thickness calvarial bone grafts were harvested from 40 rabbits and contaminated with different concentrations (range = 10(7)-10(11) colony-forming units {CFUs}/mL) of S . aureus by immersion . One graft from each rabbit was cultured immediately, and the other was inset into a calvarial donor site, fixed in place with wire, and observed for evidence of infection . At harvest (28 days postoperatively), a bacterial exposure of greater than 10(8) CFUs/mL of S . aureus was required to induce infections in the rabbit calvarial grafts (p < 0.001, Student's t-test), transmitting an infectious dose of approximately more than 10(4) CFUs per graft . These results validate the previously known dogma regarding the number of organisms needed to manifest infection {1,2} and highlight a multifold increment in bacterial count needed to transmit this infectious dose by superficial contamination alone . Such a model may be useful in investigating the efficacy of various treatment modalities of contaminated bone.

Scand J Plast Reconstr Surg Hand Surg, 1994 Sep, 28(3), 225 - 30
Persistent irritation of the soft tissue around an osseointegrated titanium implant . Case report; Holgers KM et al.; Percutaneous implants for hearing aids which are anchored in the bone have a low incidence of skin irritation . We report here the case of a man born in 1958 who had a long period of skin irritation, which was on two occasions associated with bacterial infection with Staphylococcus aureus . Despite local treatment with steroid/antibiotic ointment and skin grafts, episodes of skin irritation recurred, often in association with a relapse of ear drainage . Tests of delayed hypersensitivity to the implant and standard allergens were negative . Morphological and immunohistochemical analysis of the skin after removal of the abutment revealed an intense inflammatory reaction, with polymorphonuclear leucocytes, macrophages, lymphocytes, and HLA-DR expressing cells . In contrast to the inflammatory reaction in the soft tissue, there was a large amount of bone and the degree of bone-implant contact was good.

Semin Respir Infect, 1994 Sep, 9(3), 199 - 206
Methicillin-resistant Staphylococcus aureus as a cause of community-acquired pneumonia--a critical review; Johnston BL; Methicillin-resistant Staphylococcus aureus (MRSA) has been recognized as a nosocomial pathogen in Europe and North America for 3 decades . More recently it has emerged as a problem in long-term care facilities . It is less frequently considered a pathogen in nonfacility, community-acquired infections, where it is most often seen in intravenous drug users . There are no studies in the literature specifically describing the clinical features of MRSA pneumonia . Presumably its presentation and outcome are similar to that of pneumonia caused by susceptible strains . Staphylococcus aureus pneumonia is more often nosocomial- or nursing home-acquired, has a variable association with influenza, has clinical and laboratory features similar to other types of community-acquired pneumonia and carries a relatively high mortality of 20% to 84% . MRSA should be considered resistant to all classes of beta-lactam (beta) antibiotics . In addition, these isolates are frequently resistant to a number of other antibiotics, with vancomycin and only antibiotic to have consistently shown activity against MRSA . Therefore, vancomycin remains the treatment of choice for infections caused by MRSA, although treatment failures have been reported . The use of alternative antibiotics should be based on results of susceptibility testing of the strain isolated from the patient.

J Antimicrob Chemother, 1994 Sep, 34(3), 363 - 70
The accumulation of fluoroquinolones in Staphylococcus aureus during the postantibiotic effect; Davidson RJ et al.; The accumulation of radiolabelled pefloxacin, norfloxacin, and lomefloxacin was determined in Staphylococcus aureus during log phase growth, the postantibiotic effect (PAE), and at 1, 4 and 6 h post PAE . Uptake in actively growing cells was consistent with a non-saturable passive diffusion process . The addition of metabolic inhibitors significantly increased accumulation consistent with the presence of an energy dependent efflux system . Accumulation of all three fluoroquinolones in PAE phase cells was significantly increased compared to that which occurred in actively growing cells . In contrast to cells in the log phase, addition of metabolic inhibitors to PAE phase cells did not result in a significant increase in drug accumulation . Uptake kinetics 1 h and 4 h post PAE were similar to those found during the PAE phase . Organisms required 6 h after the end of the PAE phase before normal uptake kinetics of fluoroquinolones resumed.

Br Poult Sci, 1994 Sep, 35(4), 519 - 26
Microbiological and shelf life assessment of chilled eviscerated whole chicken broilers in Saudi Arabia; al-Mohizea IS et al.; 1 . The microbiological quality and shelf life of chicken carcasses marketed in Riyadh, Saudi Arabia were assessed . 2 . The mean initial microbial counts (log10 count/cm2) were 4.67, 4.14, 2.21, 2.78 and 2.96 for total aerobes, psychrotrophs, coliforms, Staphylococcus aureus and yeasts and moulds, respectively; these counts suggest a moderate level of contamination during processing . 3 . Yeasts and moulds were present in relatively large numbers and constituted a considerable portion of the spoilage flora . 4 . The mean shelf life of chicken broilers was 9.6, 6 and 4.4 d at 4, 7 and 10 degrees C, respectively . Storage at 4 degrees C resulted in better keeping than storage at 7 degrees C or 10 degrees C, while there was no significant difference between 7 degrees and 10 degrees C . 5 . The initial total of aerobes, psychrotrophs and yeasts and moulds were found to negatively correlate with shelf life.

Immunology, 1994 Sep, 83(1), 133 - 9
Pulmonary surfactant inhibits monocyte bactericidal functions by altering activation of protein kinase A and C; Geertsma MF et al.; Pulmonary surfactant, the main function of which is to reduce surface tension in the alveoli, is also known to affect the functions of monocytes . Two protein kinases play a role in the regulation of the bactericidal functions of phagocytes, i.e . cAMP-dependent protein kinase A (PKA), which is involved in inhibition, and Ca2+/phospholipid-dependent PKC, which is involved in stimulation of these functions . In the present study we investigated whether altered activation of PKA and/or PKC plays a role in the surfactant-induced inhibition of both the intracellular killing of Staphylococcus aureus and the production of reactive oxygen intermediates (ROI) by monocytes . The significance of increased activation of PKA was demonstrated by the following findings . Firstly, surfactant induced a sustained increase in the intracellular cAMP concentration in monocytes . Secondly, dibutyryl-cAMP (db-cAMP), a membrane-permeable cAMP analogue, mimicked the inhibitory effects of surfactant on both the killing capacity and the production of ROI by monocytes . Thirdly, an inhibitor of PKA partially restored the impaired bactericidal functions of monocytes incubated with surfactant . The involvement of decreased activation of PKC in the impaired bactericidal functions of monocytes incubated with surfactant was evident from two findings . Firstly, surfactant attenuated the phorbol myristate acetate (PMA)-mediated translocation of PKC . Secondly, surfactant inhibited the production of O2- by monocytes upon stimulation with PMA . Therefore, the mechanism involved in the surfactant-induced inhibition of the bactericidal functions of monocytes comprises both activation of an inhibitory pathway, which includes cAMP and PKA, and inactivation of a stimulatory pathway, in which PKC is involved.

Med Hypotheses, 1994 Sep, 43(3), 172 - 6
The role of immunoglobulin binding factors in the pathogenesis and therapy of AIDS; Cowan FM et al.; The human immunodeficiency virus (HIV) gp120 and gp41 envelope proteins and Staphylococcus aureus protein A (SPA) all have Fc receptor (FcR)-like immunoglobulin binding factor (IBF) activity for the Fc constant fragments of human immunoglobulin G (IgG) . Viral IBF may contribute to the pathology of HIV by jamming the network of FcR signals that control FcR-dependent immunity . Conversely, the bacterial IBF SPA has anti-retroviral activity that may involve antagonism of the immunopathological action of viral IBF, strongly suggesting IBF may act as a double-edged sword that might be turned against viral invaders.

J Clin Microbiol, 1994 Sep, 32(9), 2081 - 7
Spread and maintenance of a dominant methicillin-resistant Staphylococcus aureus (MRSA) clone during an outbreak of MRSA disease in a Spanish hospital; Dominguez MA et al.; It was not until November 1989 that the 1,000-bed University-affiliated Hospital de Bellvitge "Princeps d'Espanya" in Barcelona first acquired methicillin-resistant Staphylococcus aureus (MRSA) . Since that time, the outbreak of MRSA disease has continued . We have analyzed by genomic DNA fingerprinting 189 MRSA isolates collected between late 1989 and the end of 1993 . The isolates include both invasive and colonizing strains as well as isolates from health-care workers and environmental sources . In addition, 52 clinical isolates of methicillin-susceptible S . aureus (MSSA) collected in the same hospital were also analyzed . Isolates were classified into clonal types on the basis of molecular typing techniques . A single MRSA clone (I::B::a) belonging to ClaI type I, pulsed-field gel electrophoretic pattern B, and Tn554 pattern a was responsible for the great majority of infections (73% of blood cultures and 79% of specimens from other clinical sources) . This clone appeared at the very beginning of the outbreak, spread throughout the hospital wards, and was also carried by inpatients and health-care workers and on environmental surfaces . In contrast, no dominant lineage was apparent among MSSA isolates (33 distinct pulsed-field gel electrophoretic patterns among 52 isolates) . Two MSSA isolates seem to have originated from the dominant clone by deletion of the mecA gene and some additional DNA . In several isolates, different mecA polymorphs were present in identical chromosomal backgrounds or cells with distinct chromosomal backgrounds carried the same mecA polymorph, suggesting horizontal transfer of the mecA gene.

Ann Vasc Surg, 1994 Sep, 8(5), 500 - 5
Iliac aneurysm in a child complicating umbilical artery catheterization; Lucas A et al.; We report a case of an iliac artery aneurysm in a 9-year-old girl due to Staphylococcus aureus infection of an umbilical arterial catheter inserted at birth . After resection-anastomosis of the iliac artery the postoperative course was uneventful and hemodynamic data remain normal at 2 years . A review of the literature indicates that the prognosis of infective aortoiliac aneurysm is poor in children, except when the aneurysm is located solely in the iliac artery . Arterial continuity should be restored as late as possible after infection using a technique consistent with normal growth of the artery in the child.

Antimicrob Agents Chemother, 1994 Sep, 38(9), 2191 - 3
Failure of time-kill synergy studies using subinhibitory antimicrobial concentrations to predict in vivo antagonism of cephalosporin-rifampin combinations against Staphylococcus aureus; Brandt CM et al.; Results of in vitro time-kill synergy studies using subinhibitory, inhibitory, or suprainhibitory concentrations of bactericidal agents were compared with treatment outcomes of experimental infective endocarditis due to a methicillin-susceptible strain of Staphylococcus aureus . For rifampin-cephalosporin combinations, in vitro synergy testing using recommended fractions of the MIC failed to predict antagonism in vivo while concentrations above the MIC corresponded with antagonism in vivo.

Antimicrob Agents Chemother, 1994 Sep, 38(9), 2034 - 40
Bactericidal activities of teicoplanin, vancomycin, and gentamicin alone and in combination against Staphylococcus aureus in an in vitro pharmacodynamic model of endocarditis; McGrath BJ et al.; We adapted an in vitro pharmacodynamic model of infection to incorporate simulated endocardial vegetations . The bactericidal activities of teicoplanin, vancomycin, gentamicin, and various combinations of these drugs were studied against a strain of methicillin-susceptible Staphylococcus aureus obtained from a patient being treated for endocarditis at Detroit Receiving Hospital . Bacteria were grown overnight, concentrated, and added to a mixture of cryoprecipitate (80%) and thrombin (10%) to achieve approximately 5 x 10(9) CFU/g . Fibrin clots (8 to 10) were suspended into the model, removed at 24, 48, and 72 h in duplicate, weighed, and homogenized in 1.25% trypsin . Control experiments were conducted to characterize the growth kinetics . The following antibiotics were administered to simulate the pharmacokinetics of the drugs in humans: teicoplanin at 3 and 15 mg/kg of body weight, vancomycin at 15 mg/kg, and gentamicin at 1 mg/kg . Fibrin clot samples used to detect resistance were plated on antibiotic-containing tryptic soy agar plates . For the teicoplanin and vancomycin regimens, protein binding to cryoprecipitate, thrombin, and fibrin clot was determined to be 32, 43, and 50% and 26, 28, and 29%, respectively . In comparison with no treatment, vancomycin or teicoplanin at 15 mg/kg or either of these regimens combined with gentamicin significantly reduced bacterial counts (P < 0.0001) . Monotherapy with teicoplanin at 3 mg/kg or gentamicin resulted in no killing activity . Combination treatment with teicoplanin at 3 mg/kg and gentamicin resulted in the killing of approximately 2 log10 CFU/g by 72 h and the development of resistance to gentamicin . The results obtained with the in vitro model of endocarditis are similar to the results reported by several investigators with the rabbit model of infective endocarditis . This unique infection model is useful for designing initial drug dosage regimens and may be predictive of drug efficacy against infective endocarditis.

Antimicrob Agents Chemother, 1994 Sep, 38(9), 2014 - 23
Quinolone resistance mutations in the DNA gyrase gyrA and gyrB genes of Staphylococcus aureus; Ito H et al.; A 6.4-kb DNA fragment containing the DNA gyrase gyrA and gyrB genes was cloned and sequenced from the quinolone-susceptible Staphylococcus aureus type strain ATCC 12600 . An expression plasmid was constructed by inserting the cloned genes into the Escherichia coli-S . aureus shuttle vector pAT19, and deletion plasmids carrying only functional gyrA and gyrB genes were derived from this plasmid . An efficient transformation system for S . aureus RN4220 was established by using these plasmids . Quinolone-resistant mutants of S . aureus RN4220 were isolated by three-step selection with quinolones . The first- and second-step mutants were considered to be transport mutants, and the third-step mutants were divided into five groups with respect to their resistance patterns and transformation results with gyrA and gyrB genes . Sequencing analysis of the resulting mutant gyrase genes showed that they had the following point mutations: group 1, Ser-84 (TCA) to Leu (TTA) in GyrA; group 2, Ser-84 (TCA) to Ala (GCA), Ser-85 (TCT) to Pro (CCT), or Glu-88 (GAA) to Lys (AAA) in GyrA; group 3, Asp-437 (GAC) to Asn (AAC) in GyrB; group 4, Arg-458 (CGA) to Gln (CAA) in GyrB; and group 5, Ser-85 (TCT) to Pro (CCT) in GyrA and Asp-437 (GAC) to Asn (AAC) in GyrB . When the gyrA and/or gyrB mutants were transformed with the wild-type gyrA and/or gyrB plasmids, they became quinolone susceptible, but transformants with the plasmids having the same mutations on the gyrA and/or gyrB genes did not confer susceptibility . These results indicate that mutations in both gyrA and gyrB can be responsible for quinolone resistance in S . aureus.

Antimicrob Agents Chemother, 1994 Sep, 38(9), 1991 - 6
Characterization of mutations in Mycobacterium smegmatis involved in resistance to fluoroquinolones; Revel V et al.; Fluoroquinolone-resistant mutants of Mycobacterium smegmatis have been obtained in vitro by using ofloxacin as a selecting agent . Two types of mutants were identified according to their quinolone resistance patterns . Type 1 showed a low level of resistance to ofloxacin (MIC of 8 micrograms/ml), whereas a high level of resistance to this drug (MICs of 32 to 64 micrograms/ml) characterized type 2 . By using two oligonucleotide primers homologous to DNA sequences flanking the quinolone resistance-determining region (QRDR) in the gyrA gene of Escherichia coli and Staphylococcus aureus, a 150-bp DNA fragment was obtained by PCR amplification from total DNA of two wild-type and five mutant strains of M . smegmatis . The nucleotide sequences of the amplified fragments were determined . The deduced amino acid sequence from the wild-type strains showed ca . 79% similarity with the QRDR in the gyrase A subunit from other gram-positive and gram-negative bacteria . The DNA sequences obtained from the fluoroquinolone-resistant mutants of M . smegmatis exhibited nucleotide modifications compared with the wild-type QRDR . The QRDR from type 1 mutants had a C-T or an A-G transition leading to a change from Ala-83 to Val or Asp-87 to Gly, respectively . The QRDR from type 2 mutants had a Val-83 mutation or both Val-83 and Gly-87 mutations detected in the type 1 mutants . These results suggest that point mutations in the QRDR of the mycobacterial gyrA gene are responsible for acquired quinolone resistance in M . smegmatis.

Res Immunol, 1994 Sep, 145(7), 541 - 52
Effects of pH or ionic strength on the reactivities of mono- and polyspecific IgG antibodies; Labrousse H et al.; The reactivities of mono- and polyspecific mouse monoclonal antibodies (mAb) were compared by ELISA using immobilized antigens under different conditions, varying the pH or the NaCl concentration . The monospecific group was composed of 6 IgG directed against Staphylococcus aureus capsular polysaccharides and of 3 IgG specific to actin, myosin or tubulin . These antibodies were compared with 6 polyreactive mAb also of the IgG isotype . Marked differences were noted between the reactivities of the mono- and polyreactive IgG . pH variations had little or no effect on the reactivity of monospecific mAb to polysaccharides or to proteins . In contrast, the binding of polyreactive mAb was dependent on the pH, and the profile differed for each antigen . The NaCl concentration had opposite effects on mono- and polyreactive mAb: the binding of almost all the monoreactive mAb was increased at high NaCl concentration, while it was decreased for polyreactive mAb . In contrast, the effects of varying the pH or ionic strength on the coated antigens were negligible . The variation coefficients calculated for the pH and NaCl concentration were higher for the polyreactive mAb under study, which seems to indicate that electrostatic interactions and charged residues might be more important for these mAb than for the monoreactive ones . This characteristic might be one explanation for the particular properties of these polyreactive antibodies.

Res Immunol, 1994 Sep, 145(7), 517 - 31
Superantigen-reactive human T cells express a biased repertoire of T-cell receptor V beta joining regions; Quiros Roldan E et al.; A major characteristic of superantigens is their ability to stimulate T cells based predominantly on the type of variable segment of the T-cell receptor (TCR) V beta chain . Recently, however, reports from several laboratories have also implied a role for non-V beta elements in superantigen binding . The goal of the present study was to determine whether TCR V beta-D beta-J beta joining sequences may influence the interaction of superantigens with their target cells . To ascertain how the actual TCR repertoire of superantigen-triggered cells deviates from the theoretical one, we generated a large panel of joining region sequences from TCR carrying the TCR V beta 12 and TCR V beta 5,1 regions . The 245 sequences analysed represent transcripts of T cells from the same donor triggered either with an anti-CD3 monoclonal antibody or with the Staphylococcus aureus enterotoxins . Comparison of the joining sequences of these different groups demonstrates a skewed J beta usage in the sequences derived from superantigen-triggered cells and also provides evidence that ascribes to the putative CDR3 region of V beta segments a role in superantigen recognition . Finally, the data presented give some hints of the regions of the putative CDR3 loop that may play a major role in this function.

Lik Sprava, 1994 Sep-Dec, (9-12), 161 - 5
{The lysoamidase cleaning of the nasal and throat mucosa and hands of S . aureus carriers}; Malanchin IN et al.; A combined method has been devised of treatment of those harboring Staphylococcus aureus, using lysoamidase and myramistin . Nasopharyngeal mucosa is irrigated with buffered solution of lysoamidase from an atomizing device, the skin of the hands is treated with 0.01% solution of myramistin . A total of 138 persons have been sanitized, among whom 70 were of the medical staff of the obstetric gynecologic in-patient department, 16 doctors and nurses from the surgery department, 22 students and pedagogues from the medical institute, 30 workers of the engineering plant . The suggested method ensures efficient elimination of Staphylococcus aureus from all three biotypes in different groups for time ranges of 15 to 30 days . It can be used at health care institutions for sanitation of medical personnel, and at engineering plants in prophylaxis of purulent diseases of the hands and fingers in those workers having sustained a minor injury related to work at the plant.

Microb Pathog, 1994 Sep, 17(3), 187 - 201
Recovery of human fibroblasts from attack by the pore-forming alpha-toxin of Staphylococcus aureus; Walev I et al.; When applied at low concentrations (< 10 micrograms/ml), staphylococcal alpha-toxin generates a small channel in keratinocyte and lymphocyte membranes that permits selective transmembrane flux of monovalent ions . Here we show that a moderate concentration (1-50 micrograms/ml) of alpha-toxin similarly produces a small pore in membranes of human fibroblasts . This process leads to rapid leakage of K+ and to a drop in cellular ATP to 10-20% of normal levels in 2 h . In the presence of medium supplemented with serum and at pH 7.4, the cells are able to recover from toxin attack, so that normal levels of K+ and ATP are reached after 6-8 h at 37 degrees C . The repair process is dependent on the presence of serum in the medium and is very sensitive towards pH . Decreases of pH in the medium to < or = 7.0 as well as increases to > or = 7.8 causes the repair mechanism to fail . The fate of cell-bound toxin molecules was investigated by using a radiolabelled tracer and by immunological detection of toxin exposed at the cell surface . The results indicated that 50-70% of the toxin was shed from cell membranes . However, there was no clear correlation between shedding and recovery, and shedding was also observed in cells that died at pH 7.8 . Shedding was not decisive for repair, since cells that had recovered from toxin attack continued to carry 30-40% of initially bound toxin on their cell surface . Blockade of Na+/K(+)-ATPases with ouabain evoked similar kinetics of K(+)-depletion in control cells, compared with cells that had just recuperated from toxin attack and that still carried 30-40% alpha-toxin on their surface . We therefore tentatively concluded that repair of alpha-toxin lesions was due to closure of small pores, rather than from compensation of membrane leaks by up-regulation of Na+/K(+)-ATPase activity . We speculate that repair of small membrane lesions may extend to other agents that produce channels of similar nature in nucleated cells . Larger pores created by E . coli hemolysin or streptolysin O, both of which form larger functional transmembrane lesions, could not be repaired by fibroblasts.

Hiroshima J Med Sci, 1994 Sep, 43(3), 87 - 92
Susceptibility of methicillin-resistant Staphylococcus aureus clinical isolates to various antimicrobial agents . IV . Aminoglycoside-modifying enzyme AAC(6')/APH(2") is responsible for arbekacin-resistance enhanced by bleomycin; Inouye Y et al.; Resistance patterns against various antimicrobial agents including beta-lactams, aminoglycosides, tetracyclines, fluoroquinolones, macrolides were examined for 58 strains of methicillin-resistant Staphylococcus aureus (MRSA) isolated at Hiroshima University Hospital from April to November 1992 . All the MRSA strains produced type II-coagulase but not beta-lactamase . Regarding aminoglycoside-modifying enzymes, 7 strains (12%) appeared to be producing aminoglycoside 4',4"-adenyltransferase AAD(4',4") encoded by aadD without coproduction of bifunctional aminoglycoside 6'-acetyltransferase/2"-phosphotransferase AAC(6')/APH(2") encoded by aacA-aphD (referred to as tobramycin-resistant type, TOBr) . The remaining 51 strains (88%) were phenotypically producers of both enzymes (i.e., mix-resistant type, Mixr) . AAD(4',4"), encoded by aadD which was reported to be closely linked with bleomycin (BLM)-resistance determinant, could be seen in 100% MRSA strains and ca . 90% strains expressed AAC(6')/APH(2") . BLM endowed Mixr-type but not TOBr-type MRSA strain with enhanced resistance to arbekacin (ABK) dose-dependently, presumably by modifying the production of AAC(6')/APH(2") . The manifestation of ABK-resistant phenotype by Mixr-type MRSA required the coexistence of BLM . Therefore, ABK must be administered carefully to cure MRSA infection in patients who have been treated with BLM.

J Med Chem, 1994 Aug 19, 37(17), 2791 - 6
Synthesis and antibacterial activity of thiazolopyrazine-incorporated tetracyclic quinolone antibacterial agents . 2; Jinbo Y et al.; A novel series of 8-(2-substituted morpholino)-9,1-{(N-methylimino) methano}-7-fluoro-5-oxo-5H-thiazolo{3,2-a}quinoline-4-carboxylic acids, designated 8a-j, with a unique tetracyclic structure were synthesized, and the in vitro and in vivo antibacterial activities against Gram-positive strains, including methicillin-resistant Staphylococcus aureus isolates (MRSA), and Gram-negative strains were evaluated . These morpholino derivatives, 8a-j, showed excellent in vitro antibacterial activities against Gram-positive bacteria . The substitutions at the C-2 position of the 8-morpholino moiety of compound 8 play an important role in the enhancement of in vivo antibacterial activity . The unsubstituted morpholino derivative 8a, the 2,6-dimethyl derivative 8c, and the 2-ethylmorpholino derivative 8d showed poor in vivo antibacterial activity, while 8b, 8f-h, and 8j exhibited good activities . The 2-(methoxymethyl)morpholino derivative, 8h, showed the most potent activity in vivo . The therapeutic effects of 8h on systemic infection against S . aureus IID 803 were over 10-fold more potent than that of ofloxacin . Compound 8h, which showed superior oral bioavailability, has a chiral center . The enantiomers of 8h were synthesized, and the in vitro and in vivo antibacterial activities were evaluated . Both enantiomers, (S)-8h and (R)-8h, and the racemic compound 8 exhibited similar activities in vitro and in vivo . Compounds 8b and 8f-h also showed good levels of antibacterial activity against MRSA strains . The morpholino derivatives with unique tetracyclic structures are characterized by strong antibacterial activities against MRSA strains.

Eur J Biochem, 1994 Aug 15, 224(1), 229 - 40
Evidence that two non-overlapping high-affinity calmodulin-binding sites are present in the head region of synapsin I; Goold R et al.; Calmodulin is an important element in the regulation of nerve terminal exocytosis by Ca2+ . Calmodulin has been shown to interact with the synaptic vesicle phosphoproteins synapsins Ia and Ib {Okabe, T . & Sobue, K . (1987) FEBS Lett . 213, 184-188; Hayes, N . V . L., Bennett, A . F . & Baines, A . J . (1991) Biochem . J . 275, 93-97} . These proteins are thought to provide regulated linkages between synaptic vesicles and cytoskeletal elements . It is well established that calmodulin modulates synapsin I activities via calmodulin-dependent protein-kinase-II-catalysed phosphorylation . The direct binding of calmodulin to synapsin I suggests a second mode of regulation in addition to phosphorylation . In this study, we present evidence indicating that two sites for calmodulin binding exist in the N-terminal head region of synapsins Ia and Ib . In unphosphorylated synapsin I, these sites had a Kd value of = 36 +/- 14 nM for binding to calmodulin labelled with acetyl-N'-(5-sulpho-1-naphthyl)ethylene diamine . The Kd values for synapsin I phosphorylated at various sites were as follows: site I 18 +/- 11 nM; sites II and III 35 +/- 14 nM; sites I-III 16 +/- 9 nM . The fluorescence data indicated a stoichiometry of not less than 2 mol calmodulin bound to 1 mol synapsin I at saturation in each case . Consistent with this stoichiometry, two chemically cross-linked species (96 kDa and 116 kDa) containing calmodulin and synapsin I were generated in vitro, corresponding to one and two calmodulin molecules bound/synapsin I . Defined fragments of synapsin I were generated with the reagent 2-nitro-5-thiocyanobenzoic acid, which cleaves at cysteine residues . Cysteine-specific cleavage of whole synapsin I after cross-linking to biotinylated calmodulin generated a pair of polypeptide complexes (approximately 46 kDa and 38 kDa), the masses of which indicated cross-linking of calmodulin to the N-terminal and middle regions of synapsin I . Purified N-terminal and middle fragments each showed a Ca(2+)-dependent interaction with calmodulin affinity columns . Two calmodulin-binding fragments (7.4 kDa and 6.5 kDa) were generated using Staphylococcus aureus V8 protease digestion of synapsin I . These fragments were isolated by calmodulin affinity chromatography and reverse-phase HPLC . N-terminal sequence analysis indicated that each was contained within one of the 2-nitro-5-thiocyanobenzoic-acid-derived calmodulin-binding fragments.(ABSTRACT TRUNCATED AT 400 WORDS)

Int J Cancer, 1994 Aug 15, 58(4), 474 - 9
Progressive B-cell chronic lymphocytic leukaemia frequently exhibits aberrant p53 expression; Aguilar-Santelises M et al.; We have analysed the p53 status in non-progressive and progressive chronic B-cell leukemia (B-CLL) by ELISA, immunoprecipitation, FACS and cDNA sequencing in relation to in vitro proliferation in response to Staphylococcus aureus strain Cowan I (SAC) and IL-2 . In FACS, cells from progressive leukaemia were found to over-express p53 with a conformation recognized by PAB240 . In a PAb240-based ELISA, 60% of progressive B-CLL were positive . DNA sequencing of p53 exons 5 to 9 revealed a codon 179 His to Gln change in one of the ELISA-positive, progressive B-CLL but failed to reveal any mutations in 4 other ELISA-positive, progressive B-CLL . Among progressive B-CLL populations, 10/14 responded by proliferation in vitro to SAC/IL-2 . In non-progressive cells, low levels of p53 were found by FACS, none was positive in the PAb240 ELISA and only one case showed a weak proliferative response to SAC/IL-2 . Low p53 expression was also seen in different types of normal B cells, resting and activated, and in EBV-transformed B-cell lines, in contrast to the high expression observed in Burkitt lymphoma cell lines with verified p53 mutations . We conclude that progressive B-CLL is characterized by aberrant p53 expression which may be a significant prognostic factor.

J Immunol, 1994 Aug 15, 153(4), 1466 - 77
Differential regulation of C alpha 1 and C alpha 2 germ-line and mature mRNA transcripts in human peripheral blood B cells; Kitani A et al.; In this study, we investigated the regulation of germ-line and mature C alpha mRNA transcript expression in human peripheral blood B cells . In initial studies, we found that C alpha germ-line and mature transcripts were constitutively expressed in total peripheral blood B cells but not in high density surface IgM+ (surface IgA-) B cells; the latter cells were therefore used in subsequent studies . Focusing first on regulation of germ-line (I alpha-C alpha) transcripts, we showed that C alpha 1 germ-line transcripts are induced by TGF-beta 1 alone but such induction is enhanced by Staphylococcus aureus, Cowan I (SAC) . In contrast, C alpha 2 germ-line transcripts are optimally induced by TGF-beta 1 in the absence of a B cell stimulus . In addition, although SAC alone induced both C alpha 1 and C alpha 2 germ-line transcripts, such induction is dependent on endogenous (B cell-derived) TGF-beta 1 production, because no induction is detected in cells cultured in the presence of neutralizing anti-TGF-beta 1 . Turning next to mature (VDJ-C alpha) transcripts we showed that SAC plus TGF-beta 1 induces mature C alpha 1 transcripts, and such induction is IL-10 dependent because it is enhanced by exogenous IL-10 and abrogated by the presence of anti-IL-10 . In contrast, SAC plus TGF-beta 1 does not induce C alpha 2 mature transcripts, even in the presence of exogenous IL-10; such transcripts are induced, however, by T cell stimuli such as anti-CD40 (presented by CDw32-transfected L cells) in the presence of TGF-beta 1/IL-10 or by PWM-activated T cells . In summary, these studies show that C alpha 1 and C alpha 2 germ-line transcript induction is differentially regulated, in keeping with previous studies of differences in germ-line CH transcript induction in the first and second IgH duplication units . Furthermore, C alpha 1 and C alpha 2 mature transcript induction is also differentially regulated, with C alpha 2 requiring a T cell signal such as that delivered by the CD40 ligand . Finally, IL-10 appears to be uniquely supportive of the induction of both C alpha 1 and C alpha 2 mature transcripts.

Structure, 1994 Aug 15, 2(8), 747 - 54
The structure of an asymmetric dimer relevant to the mode of action of the glycopeptide antibiotics; Groves P et al.; BACKGROUND: Glycopeptide antibiotics of the vancomycin group are of crucial clinical importance in the treatment of methicillin resistant Staphylococcus aureus (MRSA)--the often lethal 'super-bug'--characterized by its resistance to a wide range of antibiotics in common use . The antibiotics exert their physiological action by blocking cell wall synthesis through recognition of nascent cell wall mucopeptides terminating in the sequence -D-Ala-D-Ala . Evidence suggests that the antibiotics are able to enhance their biological activity by the formation of homodimers, and this is supported by the observation that dimerization and peptide binding in vitro are cooperative phenomena . The basis of this enhancement is not understood at the molecular level . RESULTS: The first detailed structure of a dimeric glycopeptide antibiotic, that of eremomycin, is presented based upon solution NMR data . The overall structure of the dimer complex is asymmetric . The source of this asymmetry--a parallel alignment and mutual interaction of the disaccharides--appears to promote dimerization through specific sugar-sugar recognition . CONCLUSIONS: A molecular basis for the observed cooperativity of cell wall peptide binding by eremomycin is evident from these studies of the dimer . The carboxylate anion of the cell wall component, which is crucial to binding, forms an amide-mediated ion-pair interaction to the alkylammonium ion of the ring 6 sugar in the other half of the dimer making the structure and positioning of this sugar important in mediating cooperativity.

Br J Nurs, 1994 Aug 11-Sep 7, 3(15), 753 - 9
Methicillin-resistant Staphylococcus aureus: do we just have to live with it?
Siu AC.
Infection with methicillin-resistant Staphylococcus aureus is a familiar problem in hospital patients and has been a global issue since the 1960s . Control of this organism is expensive both financially and in terms of human resources . This article is an attempt to review the epidemiological evolution and our understanding of the organism and bring together possible solutions.

Eur J Pharmacol, 1994 Aug 3, 270(4), 379 - 82
Septic shock: no correlation between plasma levels of nitric oxide metabolites and hypotension or lethality; van den Berg C et al.; In the Wistar rat (Riv:TOX strain), Escherichia coli-derived lipopolysaccharide, up to 100 mg/kg, did not affect blood pressure . However, 6 h after administration of live E . coli or Staphylococcus aureus (a microorganism without lipopolysaccharide), both dosed at 12 x 10(9) colony forming units/kg, mean arterial blood pressure significantly decreased to 64% and 48% compared to control, respectively . In contrast to lipopolysaccharide, bacteria produced a dose-dependent lethality within 24 h . Live S . aureus increased plasma levels of nitric oxide metabolites (NOx) only four-fold, while both lipopolysaccharide and live E . coli approximately 20-fold . In conclusion, we demonstrated a lack of correlation between plasma NOx levels and hypotension or lethality.

Laryngoscope, 1994 Aug, 104(8 Pt 1), 927 - 31
Toxic shock syndrome after functional endonasal sinus surgery: an all or none phenomenon?
Abram AC, Bellian KT, Giles WJ, Gross CW.
Reported cases of toxic shock syndrome (TSS) following nasal surgery or functional endonasal sinus surgery (FESS) are uncommon . Classic TSS is a serious multisystem disorder resulting from Staphylococcus aureus phage I toxic shock syndrome toxin 1 (TSST-1), and it is characterized by fever, rash, hypotension, mucosal hyperemia, vomiting, diarrhea, and laboratory evidence of multisystem organ dysfunction . TSS cases following nasal surgery have been associated with nasal packing, mucosal barrier violation, prior S aureus phage I colonization, as well as low antitoxin antibody levels . Of the 1700 FESS procedures performed at our institution, 3 cases were complicated by classic TSS, with 2 additional patients having a postsurgical course compromised by a milder degree of TSS . Diagnostic criteria, clinical presentation, management, and etiology are discussed, and the possibility of a continuum from mild-to-classic TSS is addressed.

J Lab Clin Med, 1994 Aug, 124(2), 293 - 301
N-acetylcysteine enhances in vitro the intracellular killing of Staphylococcus aureus by human alveolar macrophages and blood polymorphonuclear leukocytes and partially protects phagocytes from self-killing; Oddera S et al.; The processes of phagocytosis and intracellular killing of bacteria by alveolar macrophages (AMs) and polymorphonuclear leukocytes (PMNs) result in the production of reactive oxygen species that can induce self-damage to the phagocytic cells . N-Acetylcysteine (NAC), a mucolytic agent used to treat chronic respiratory inflammatory disorders, possesses antioxidant properties and has therefore been used for the prevention of damage induced by oxygen radicals . This study was designed to evaluate whether NAC can interfere with the processes of phagocytosis and intracellular killing of bacteria and protect the phagocytic cells from self-killing . Human AM, obtained by bronchoalveolar lavage, and peripheral blood PMNs were cultured with Staphylococcus aureus (American Type Culture Collection 25923 strain) in the presence of different concentrations of NAC (1, 10, and 100 mg/L) and two chromophores (4',6'-diamidino-2-phenylindole dihydrochloride and propidium iodide), which identify live or dead bacteria and dead phagocytes . As compared with PMNs, AMs were more effective in ingesting bacteria (p < 0.05) and were equally effective as intracellular killers (p > 0.05), but were susceptible to a significantly higher rate of self-killing (p < 0.01) . The presence of NAC in the cell cultures at the highest dose tested (100 mg/L) induced a significant enhancement of the bactericidal activity of both AMs (p < 0.05) and PMNs (p < 0.05) . This increased intracellular killing was not associated with increased proportions of dead phagocytes either in AMs or PMNs cultures (p > 0.05, each comparison), suggesting a protective effect of NAC on damage induced by toxic products generated during phagocytosis.

J Bacteriol, 1994 Aug, 176(16), 4993 - 5000
Cloning and characterization of a gene affecting the methicillin resistance level and the autolysis rate in Staphylococcus aureus; Maki H et al.; Tn918 mutagenesis of a high-level methicillin-resistant Staphylococcus aureus (methicillin MIC, 800 micrograms/ml) led to the isolation of a low-resistance mutant . The Tn918 insert was transferred back to the parent to produce strain SRM563 (methicillin MIC, 12.5 micrograms/ml), which showed heterogeneous resistance . Twenty-two clinical isolates of methicillin-resistant S . aureus were transformed with DNA of SRM563 . In the transformants of most strains, instances of reduced resistance were observed with concomitant increases of autolysis rate induced by Triton X-100 and were generally more profound in high-resistance strains . Two transformants exhibited a decrease of the autolysis rate and little reduction of resistance . In the transformant of methicillin-susceptible strain RN2677, an increase of the autolysis rate and little reduction of resistance were observed . The production of low-affinity penicillin-binding protein (PBP2') did not significantly decrease in the mutants . Insertion of Tn918 occurred within the 3'-terminal region of a novel gene designated llm, which was cloned and sequenced . RNA blot analysis demonstrated that the gene was transcribed . The encoded protein was composed of 351 amino acid residues with a molecular weight of 38,512 and was hydrophobic, suggesting its location on the membrane . The gene was detected by PCR in all S . aureus strains tested but not in the other 26 staphylococcal species . Comparison of the 3'-terminal sequences of the gene among several S . aureus strains showed that, whereas nucleotide substitutions occurred at the third position in seven of eight 3'-terminal codons, only C-terminal amino acid variation of glutamate or aspartate was observed.

J Bacteriol, 1994 Aug, 176(16), 4883 - 9
Genetic analysis of type 5 capsular polysaccharide expression by Staphylococcus aureus; Lee JC et al.; Capsules are produced by over 90% of Staphylococcus aureus strains, and approximately 25% of clinical isolates express type 5 capsular polysaccharide (CP5) . We mutagenized the type 5 strain Reynolds with Tn918 to target genes involved in CP5 expression . From a capsule-deficient mutant, we cloned into a cosmid vector an approximately 26-kb EcoRI fragment containing the transposon insertion . In the absence of tetracycline selection, Tn918 was spontaneously excised, thereby resulting in a plasmid containing 9.4 kb of S . aureus DNA flanking the Tn918 insertion site . The 9.4-kb DNA fragment was used to screen a cosmid library prepared from the wild-type strain . Positive colonies were identified by colony hybridization, and a restriction map of one clone (pJCL19 with an approximately 34-kb insert) carrying the putative capsule gene region was constructed . Fragments of pJCL19 were used to probe genomic DNA digests from S . aureus strains of different capsular serotypes . Fragments on the ends of the cloned DNA hybridized to fragments of similar sizes in most of the strains examined . Blots hybridized to two fragments flanking the central region of the cloned DNA showed restriction fragment length polymorphism . A centrally located DNA fragment hybridized only to DNA from capsular types 2, 4, and 5 . DNA from pJCL19 was subcloned to a shuttle vector for complementation studies . A 6.2-kb EcoRI-ClaI fragment complemented CP5 expression in a capsule-negative mutant derived by mutagenesis with ethyl methanesulfonate . These experiments provide the necessary groundwork for identifying genes involved in CP5 expression by S . aureus.

Infect Immun, 1994 Aug, 62(8), 3416 - 23
Platelet microbicidal protein alone and in combination with antibiotics reduces Staphylococcus aureus adherence to platelets in vitro; Yeaman MR et al.; Bacterial adherence to platelets on the cardiac valve surface is believed to be critical in the induction of infective endocarditis . Recent studies have confirmed that thrombin-activated platelets secrete platelet microbicidal protein (PMP), which can both kill and exert nonlethal antiadherence effects against endovascular pathogens . In the present study, we quantified the influence of antibiotic and/or PMP exposures on in vitro platelet adherence of two Staphylococcus aureus strains, identical by DNA restriction and cell wall protein profiles, that differed in their susceptibility to PMP-induced killing (PMPs or PMPr, respectively) . Adherence assays were performed by flow cytometry in the presence of sublethal PMP concentrations (1 to 2.5 micrograms/ml) alone or in combination with ampicillin (AMP) alone, sulbactam (SUL) alone, or AMP plus SUL (AMP-SUL), at levels achievable in serum . Exposure of the PMPs and PMPr S . aureus strains to antibiotics (for 2 h at 37 degrees C) prior to flow cytometry resulted in no substantive changes in the percent adherence to platelets compared with that for S . aureus cells not exposed to antibiotics, except for modestly increased adherence of both PMPs and PMPr cells exposed to AMP-SUL (18.5 and 15.8% increases, respectively) . Addition of PMP to antibiotic-S . aureus mixtures (final 30 min) caused a significant decrease in S . aureus adherence to platelets, for both the PMPs and PMPr S . aureus strains, compared with antibiotic exposure alone (e.g., reduction in platelet adherence from 57.9 +/- 8.2% to 12.2 +/- 3.6% for PMPs cells exposed to AMP-SUL and PMP {P = 0.01}) . Moreover, addition of PMP following exposure of the PMPs and PMPr strains to AMP-SUL reversed the enhanced bacterium-platelet adherence observed with such antibiotic exposures alone (P < or = 0.005) . These data demonstrate that PMP exerts a potent antiplatelet adherence effect which is independent of its microbicidal capacity, rendering S . aureus cells less adherent to platelets in the presence or absence of antibiotics . Reduction of microbial adherence to platelets by PMP alone or with antibiotics provides further insight into the mechanism(s) that may be involved in host defense and antibiotic prophylaxis of infective endocarditis and other endovascular infections.

Infect Immun, 1994 Aug, 62(8), 3184 - 8
Interaction of the two components of leukocidin from Staphylococcus aureus with human polymorphonuclear leukocyte membranes: sequential binding and subsequent activation; Colin DA et al.; The sequential interaction between the two components S and F of leukocidin from Staphylococcus aureus and the membrane of human polymorphonuclear neutrophils has been investigated in the presence of 1 mM Ca2+ . With 125I-labeled components, it has been shown that binding of the F component occurred only after binding of the S component . The kinetic constants of binding of both components were not statistically different (Kd, approximately 5 nM; Bm, approximately 35,000 molecules per cell), and both Hill coefficients were 1 . The application of increasing concentrations of leukocidin provoked a dose-dependent secretion of the granule content, as determined by hexosaminidase and lysozyme activity measurements . Furthermore, the separate perfusion of S and F components on human polymorphonuclear neutrophils deposited on a filter induced secretion of the granules content only when the perfusion of the S component preceded that of the F component . We conclude, therefore, that (i) S-component binding is a prerequisite for F-component binding and for subsequent activation of polymorphonuclear neutrophils and (ii) there is a specific binding site for the S component in the plasma membrane.

Radiology, 1994 Aug, 192(2), 363 - 5
Infection with fatal outcome after endovascular metallic stent placement; Therasse E et al.; The authors describe a case in which infection developed in a patient after placement of a metallic endoprosthesis in the iliac artery . The patient died of respiratory failure secondary to sepsis . Imaging studies did not reveal an abscess, aneurysm, or stent occlusion . Stent cultures and hemocultures grew Staphylococcus aureus . As with any foreign body, infectious complications may occur after insertion of metallic stents and should be recognized as soon as possible to initiate appropriate therapy.

J Clin Microbiol, 1994 Aug, 32(8), 1866 - 9
Development of enzyme-labeled oligonucleotide probe for detection of mecA gene in methicillin-resistant Staphylococcus aureus; Shimaoka M et al.; A DNA hybridization method with an enzyme-labeled oligonucleotide probe (mecA-ELONP) was developed to detect the methicillin-resistant gene (mecA) in methicillin-resistant Staphylococcus aureus . For rapid identification, bacterial colonies were transferred from agar plates directly onto nylon membranes . Lysis of cells, denaturation of DNA, and hybridization were performed on the membranes . These procedures required only 3 h for completion . The results obtained by this test closely corresponded with those obtained by determining the MICs of oxacillin against S . aureus . The results of the mecA-ELONP also correlated well with those of a commercially available PCR test . Thus, mecA-ELONP proved to be a reliable and convenient method for the rapid identification of methicillin-resistant S . aureus, which could be useful in clinical microbiology laboratories.

Antimicrob Agents Chemother, 1994 Aug, 38(8), 1703 - 10
Teicoplanin alone or combined with rifampin compared with vancomycin for prophylaxis and treatment of experimental foreign body infection by methicillin-resistant Staphylococcus aureus; Schaad HJ et al.; The prophylactic and therapeutic activities of teicoplanin were evaluated in two different experimental models of foreign body infections caused by methicillin-resistant Staphylococcus aureus (MRSA) . In a guinea pig model of prophylaxis, subcutaneously implanted tissue cages were infected at a > 90% rate by 10(2) CFU of MRSA in control animals . A single dose of 30 mg of teicoplanin per kg of body weight administered intraperitoneally 6 h before bacterial challenge was as effective as vancomycin in preventing experimental infection in tissue cages injected with either 10(2), 10(3), or 10(4) CFU of MRSA . In a rat model evaluating the therapy of chronic tissue cage infection caused by MRSA, the efficacy of a 7-day high-dose (30 mg/kg once daily) regimen of teicoplanin was compared with that of vancomycin (50 mg/kg twice daily) . Whereas high levels of teicoplanin were found in tissue cage fluid, continuously exceeding its MBC for MRSA by 8- to 16-fold, no significant reduction in the viable counts of MRSA occurred during therapy . In contrast, either vancomycin alone or a combined regimen of high-dose teicoplanin plus rifampin (25 mg/kg twice daily) could significantly decrease the viable counts in tissue cage fluids . Whereas the bacteria recovered from tissue cage fluids during therapy showed no evidence of teicoplanin resistance, they failed to be killed even by high levels of this antimicrobial agent . The altered susceptibility of in vivo growing bacteria to teicoplanin killing might in part explain the defective activity of this antimicrobial agent when used as monotherapy against chronic S . aureus infections . These data may indicate the need for a combined regimen of teicoplanin with other agents such as rifampin to optimize the therapy of severe staphylococcal infections.

Am J Infect Control, 1994 Aug, 22(4), 224 - 7
Comparative effectiveness of hand-cleansing agents for removing methicillin-resistant Staphylococcus aureus from experimentally contaminated fingertips; Huang Y et al.; BACKGROUND: Five subjects participated in a study of optimal conditions for removing methicillin-resistant Staphylococcus aureus from contaminated fingertips . METHODS: Fingertips were contaminated experimentally and cleaned by various methods . Bacterial removal was measured as percentage and is given as mean +/- standard error of the mean . RESULTS: Rinsing the fingertips with tap water for 20 seconds and drying them with paper towels removed 95.2% +/- 1.6% of the contamination . Application of hand-cleansing agents to fingertips for 20 seconds, followed by a 20-second tap-water rinse and towel drying, removed bacteria as follows: povidone-iodine detergent, 99.2% +/- 0.4%; chlorhexidine detergent, 97.2% +/- 0.8%; and liquid soap, 96.1% +/- 1.1% . In a modification of the method, 80% ethyl alcohol applied to the fingertips for 20 seconds, followed by air drying for 40 seconds, removed 99.1% +/- 0.8% of the bacteria . Statistical analyses indicated that povidone-iodine and 80% ethyl alcohol were more effective than the other agents . CONCLUSIONS: Rinsing contaminated fingertips with tap water and towel drying them is sufficient to reduce contamination with methicillin-resistant S . aureus by 95% . Washing with povidone-iodine or 80% ethyl alcohol further reduces contamination by an additional 99%.

Int J Pept Protein Res, 1994 Aug, 44(2), 183 - 91
Solid-phase peptide synthesis and biological activity of bovine thymopoietin II (bTP-II); Smith DD et al.; Bovine thymopoietin (bTP), a 49 amino acid polypeptide, was synthesized using Merrifield's solid-phase peptide synthesis methodology . The polypeptide was purified using anion-exchange chromatography and reversed-phase HPLC and characterized by mass spectrometry and amino acid analysis of the full-length peptide and of products derived from digestion with Staphylococcus aureus V8 protease . The biological activity of the synthesized product was tested in several assay systems . Synthetic bTP was found to induce the expression of Thy 1.2 antigen on T-lymphocytes from athymic mice, in agreement with previous studies on the biological activity of endogenous bTP . Biological activity at skeletal muscle and neuronal nicotinic acetylcholine receptor sites, as reported by others for bTP, could not be confirmed in our studies . The absence of biological activity at nicotinic receptor sites may be related to the results of a recent report demonstrating the presence of a cobratoxin-like molecule in preparations of natural bTP . These data indicate that synthetic peptides have an important role for the evaluation of the specificity of the biological activity of polypeptides.

Ann Plast Surg, 1994 Aug, 33(2), 142 - 7
Debridement of bacterial and particulate-contaminated wounds; McDonald WS et al.; Debridement of contaminated wounds is an essential component of uncomplicated wound healing . Efficient techniques should be capable of removing bacteria as well as foreign matter because of the well-known ability of foreign bodies to potentiate infection . We have compared the ability of current debridement techniques with the relatively new ultrasound debridement to clean wounds contaminated with bacteria and particulate matter . In part I, we prepared dorsal wounds on 15 Sprague-Dawley rats, and 20 mg of Montmorillonite clay soil fraction, a well-known infection-potentiating factor, was placed in each wound . Animals were randomly assigned to one of three treatment groups: ultrasound debridement, soaking, and irrigation . The amount of clay removed from each wound was measured . In part II, 48 Sprague-Dawley rats were given a standard wound and inoculated with a subinfective dose of Staphylococcus aureus bacteria and 10 mg of Montmorillonite clay particles . Each rat was randomly assigned to a debridement technique--soaking, scrubbing, high-pressure irrigation, and ultrasound--and was examined after 7 days for inflammatory responses . Results of part I demonstrated that ultrasound debridement and irrigation remove statistically equal amounts of clay (p < 0.05) . In part II, we found that high-pressure irrigation and ultrasound debridement effectively treat contaminated wounds (gross infection, p < 0.05; wound induration, p < 0.0001; bacteria counts, p < 0.002) . This study presents ultrasound debridement as effective treatment of contaminated wounds.

Vaccine, 1994 Aug, 12(11), 988 - 92
Vaccination against Staphylococcus aureus mastitis: immunological response of mice vaccinated with fibronectin-binding protein (FnBP-A) to challenge with S . aureus; Mamo W et al.; Mice were immunized with fusion proteins encompassing the fibronectin-binding domain of a staphylococcal fibronectin-binding protein (FnBP-A) . A specific antibody response against the fibronectin-binding part of the fusion proteins was detected in the serum of all vaccinated animals . The protective potential of these vaccinations was evaluated in a mouse mastitis model, using Staphylococcus aureus, strain SA113, for challenge . The mice vaccinated with FnBP fusion proteins showed a decreased number of bacteria recovered from the mammary glands and significantly reduced cases of severe mastitis . Histopathological examination of tissue from challenged glands of vaccinated mice revealed either no pathological reactions or disseminated inflammatory reactions with focal necrosis whereas four of six examined tissues from challenged glands of non-vaccinated animals showed total necrosis . A combination of FnBP fusion protein with staphylococcal alpha-toxoid did not increase the efficacy of the vaccination and animals vaccinated with alpha-toxoid alone were as sensitive to challenge as those from the non-vaccinated control group . Thus vaccination of mice with recombinant FnBP resulted in significant protection against challenge with S . aureus.

Vet Microbiol, 1994 Aug 1, 41(3), 249 - 58
Pathogenicity in mice of Staphylococcus aureus mutants deficient in exoprotein synthesis; Odierno L et al.; Twelve mutants were isolated from a Staphylococcus aureus strain derived from bovine mastitis after mutagenesis by ultraviolet light . These mutants were found to be deficient for several characteristics such as production of most exoproteins and had altered phage type and/or colonial morphology in serum-soft agar medium . They also differed in virulence when assayed in mice by intraperitoneal administration; the ratio of the LD50 of the mutants vs . that of the parental strain ranged from 1 to 123 . The different virulence of the mutants could not be associated with lack of production of exoproteins or altered colonial morphology . On the other hand, a clear correlation was evidenced between lowered virulence and slower growth rate at 37 degrees C . Three mutants were assayed in the mouse mastitis model . One of them, which was about 40 times less virulent when assayed by intraperitoneal administration, induced a histopathological lesion similar to that produced by the parent strain; the other two mutants, which were about 70 to 120 times less virulent by intraperitoneal administration, induced only a very slight lesion . Mice were vaccinated by the intraperitoneal route with two of the less virulent mutants; the LD50 in the vaccinated mice that were challenged with the parental strain increased 11 to 14 times compared with that for the unvaccinated mice.

Ryumachi, 1994 Aug, 34(4), 786 - 9
{Gluteal abscess caused by Staphylococcus aureus in a patient with systemic lupus erythematosus}; Yoshino Y et al.; We describe here a 44-year-old female patient with systemic lupus erythematosus (SLE), who developed gluteal abscess resulting in symptoms suggestive of aseptic necrosis of the right femoral head . The patient was diagnosed as active SLE with organic brain syndrome in December 1992, and has been treated with high doses of methylprednisolone (initial dose: 100 mg daily) . As she recovered from the manifestations, the dose of methylprednisolone has been gradually decreased . In June 1993, she began to complain of right leg pain on walk with positive Patrick sign in the right hip . A massive gluteal abscess was identified by X-ray and CT scan, and drained . Cultures of the purulent fluid yielded Staphylococcus aureus . Of note, Staphylococcus aureus had been detected from repeated cultures of pharyngeal swabs, suggesting that the organism invading from the pharynx was carried in the blood to a small hematoma in the right gluteal muscle to form an abscess . The importance of an awareness of the possibility of the infection of Staphylococcus aureus in SLE patients is discussed.

Rev Esp Cardiol, 1994 Aug, 47(8), 518 - 22
{Infectious endocarditis in non-addict patients without predisposing heart disease . Differential features}; Ruiz M et al.; INTRODUCTION AND AIMS . Although uncommonly, infective endocarditis in non-addict patients may involve people without predisponente heart disease . The aim of our study was to assess the clinical and prognostic features of this type of endocarditis and to compare them with those of the more common type of endocarditis with underlying lesion . METHODS . With this aim, we have reviewed 71 consecutive cases of non-addict infective endocarditis diagnosed in our hospital in the last 7 years; there was no preexisting cardiac lesion in 9 patients (13% of all endocarditis and 21% of native valve endocarditis), while underlying heart disease, including mitral valve prolapse, was present in the remaining 62 patients . RESULTS . Mean age was significantly lower in 9 patients without preexistent lesion (28 +/- 18 versus 46 +/- 17 years, p < 0.01), while there was no differences for gender . Infection involved the aortic valve in 56%, the tricuspid or pulmonary valve in 33% and the mitral valve in only 11% of the patients without underlying cardiopathy (for 44%, 4% and 49%, respectively, in patients with cardiopathy) . Staphylococcus aureus caused 67% of cases in patients without cardiopathy and only 9% in those with cardiopathy . Surgery was required in a similar proportion by both groups of patients (55% and 56%), although mortality was more than twice higher in patients with prior cardiac lesions (25% versus 11%) . CONCLUSIONS . A significant proportion of non-addict infective endocarditis involves patients without predisponente heart disease . These cases have some differential features (younger age, aortic and right heart valves involvement, S . aureus as the main causative agent and lower mortality) in comparison to those of endocarditis in patients with underlying cardiopathy.

J Hosp Infect, 1994 Aug, 27(4), 275 - 83
Changes in microbial ecology and use of cloxacillin; Loulergue J et al.; There is generally a positive correlation between antibiotic consumption and incidence of resistance to antibiotics used either for prophylaxis or therapy in human infections . This was not the case for two surgical wards in our hospital . A 15-year study showed that the incidence of methicillin-resistant Staphylococcus aureus (MRSA) was unrelated to cloxacillin consumption, and in fact fell after introduction into the two wards of an antibiotic policy based on cloxacillin . The two wards, a 90-bed orthopaedic unit and a 60-bed trauma unit, had an incidence of MRSA that has remained below the hospital average (23% in 1989, 32% in 1992) . Before introduction of the policy the incidence of MRSA in 1977-1979 in the orthopaedic ward was 31%, and in the trauma ward 33% . In 1989 an investigation revealed no MRSA carriers in staff of either ward . In contrast, seven MRSA carriers were found among staff and patients of three other surgical units selected, because the percentage of MRSA isolated was above the average in our hospital . However, a different type of patient is found in these units, the treatment techniques differ and broader-spectrum antibiotics are used . In addition to the usual precautions regarding nursing care and isolation techniques, the best means of reducing MRSA epidemics is to reduce the reservoir of carriers . The fall in the MRSA infection rate in orthopaedic and traumatology wards can be explained by antibiotic policy but also by other infection control measures.

J Dairy Sci, 1994 Aug, 77(8), 2281 - 8
Staphylococcus aureus colonization of teat skin as affected by postmilking teat treatment when exposed to cold and windy conditions; Fox LK et al.; Study 1 was conducted to determine whether postmilking teat treatment with ointment before exposure to cold and wind resulted in better skin health than standard teat treatment . Teat treatments tested were 1% I2 and 10% glycerin, ointment with 1% chloroxylenol, ointment with .3% 8-hydroxyquinoline sulfate, and no treatment (control) . Teats were treated 7 d prior to chapping . A broth culture of Staphylococcus aureus was applied once to teats after chapping was established . Treatments were applied after milking and before sample collection for 11 d following S . aureus application . Milk samples were collected aseptically, teat skin swabbing solutions were collected, and teat condition was scored . Cows were exposed to ambient winter conditions, and a wind velocity of 152.4 m/min was applied to the mammary gland surface for 15 min immediately postmilking . Ointment and control teats had significantly better skin condition than teats treated with I2 solution . Colonization of S . aureus was greatest on ointment treated teats . Study 2 was conducted to determine whether teat condition of cows receiving postmilking I2 solution treatments would be improved if teats were blotted dry before exposure to wind and cold ambient conditions . Two mammary quarters of each cow received I2 solution treatment of study 1, but teats were blotted dry prior to exit from the milking parlor . No treatment was applied to the other teats . Teat condition scores were similar between treatments, but S . aureus colonization was significantly greater on control teats . Results indicate a possible disadvantage to treating teats with ointments after milking, as evidenced by increased S . aureus colonization.(ABSTRACT TRUNCATED AT 250 WORDS)

Indian J Exp Biol, 1994 Aug, 32(8), 553 - 8
Immunotherapeutic modification by an ayurvedic formulation Septilin; Rao CS et al.; Effect of Septilin, an ayurvedic formulation proven to be effective in the therapy of chronic infections, was investigated on the phagocytic system and humoral response in rats and mice . Septilin exhibited significant protection in E . coli-induced abdominal sepsis in normal mice and in Staphylococcus aureus-induced sepsis in neutropenic mice . It significantly reduced the viable E . coli cells when incubated with neutrophils in rats . Septilin stimulated the phagocytic function of the reticuloendothelial system in mice . In normal rats, Septilin enhanced anti-SRBC hemagglutination antibody titre by 5.7-fold and showed significant protection in cyclophosphamide-induced humoral suppression.

J Dermatol Sci, 1994 Aug, 8(1), 1 - 10
Staphylococcus aureus infection on experimental croton oil-inflamed skin in mice; Akiyama H et al.; Staphylococcus aureus cells were inoculated on the surface of skin inflamed by application of croton oil in cyclophosphamide-treated mice . Skin specimens were taken at 1, 3, 6, 12, and 24 h inoculation and each specimen was examined by microscopy . The S . aureus cells which attached to the surface of the skin immediately after inoculation had invaded the horny layer within 1 h . The cells gradually penetrated deeper into the epidermis . Electron microscopy revealed fibril-like structures around the S . aureus cells and the cells which adhered to the horny layer and fibrin by means of Ruthenium red-positive, fibril-like structures . A combined application of 0.1% gentamicin ointment, 2% fusidic acid ointment, and clobetasol propionate ointment was more effective in decreasing the number of S . aureus cells in the lesions than was an application of clobetasol propionate ointment alone . However, a combined application of 0.1% gentamicin ointment and 2% fusidic acid ointment without clobetasol propionate ointment showed almost the same efficacy as that with clobetasol propionate ointment . Although povidone iodine killed S . aureus in vitro at a concentration of 0.01% (100 micrograms/ml) in 40 s, its in vivo efficacy was limited.

Planta Med, 1994 Aug, 60(4), 365 - 8
Xanthones of Garcinia cowa; na Pattalung P et al.; Five xanthones have been isolated from Garcinia cowa Roxb . (Guttiferae): cowanin (1), cowanol (2), cowaxanthone (3), 1,3,6-trihydroxy-7-methoxy- 2-5-bis(3-methyl-2-butenyl)xanthone (4), and norcowanin (5) . The structures were assigned by spectroscopic studies . Xanthones 2 and 3 showed moderate antimicrobial activity against Staphylococcus aureus.

Can J Microbiol, 1994 Aug, 40(8), 677 - 81
Characterization of a Tn551-mutant of Staphylococcus aureus defective in the production of several exoproteins; Giraudo AT et al.; A Tn551 insertional pleiotropic mutant defective in the production of several exoproteins was isolated from Staphylococcus aureus 196E and characterized . The pleiotropism of the mutant was due to a single insertion of the transposon as evidenced by Southern blot hybridization and by the transfer of its phenotype by transduction to S . aureus ISP479 . The mutants showed diminished or null levels of alpha- and beta-hemolysis, DNase, coagulase, and protein A in the supernatants of broth cultures . Production of proteases, lipase, staphylokinase, or enterotoxin A was not modified . The mutants did synthesize the cell-bound form of protein A and also the extracellular form of this protein coded by pRIT11, which lacks the COOH-terminal segment of the molecule . These observations suggest that the sae locus does not involve a positive regulatory gene acting at the transcriptional level . The phenotype of the mutant was different from that of other insertional mutants affecting exoprotein synthesis, such as agr, xpr, or sar . This new mutation has been designated sae (for S . aureus exoprotein expression).

Mol Pharmacol, 1994 Aug, 46(2), 329 - 37
Localization of the forskolin labeling sites to both halves of P-glycoprotein: similarity of the sites labeled by forskolin and prazosin; Morris DI et al.; An iodinated derivative of forskolin, 6-O-{{2-{3-(4-azido-3-{125I} iodophenyl)propionamido}ethyl}carbamyl}forskolin ({125I}6-AIPP-Fsk), photolabels the multidrug efflux pump P-glycoprotein in membranes prepared from the multidrug-resistant cell lines KB-V1 and KB-C1 . The labeling site for {125I}6-AIPP-Fsk was localized by immunoprecipitation of tryptic fragments of P-glycoprotein labeled in KB-C1 membranes . A 6-kDa, photolabeled, tryptic fragment was immunoprecipitated by antiserum raised against residues 348-419 of P-glycoprotein, PEPG9, but not by antisera raised against flanking regions PEPG7 and PEPG11 . A peptide that corresponds to residues 343-359 of P-glycoprotein inhibited immunoprecipitation of the 6-kDa fragment by antiserum against PEPG9 but had no effect on the immunoprecipitation of photolabeled fragments by antiserum against PEPG7 . A second peptide, corresponding to residues 360-376, had no effect on the immunoprecipitation by antiserum against PEPG9 . {125I}6-AIPP-Fsk labels the carboxyl-terminal half of P-glycoprotein, because low molecular mass tryptic fragments were immunoprecipitated by three carboxyl-terminal antisera . Therefore, {125I}6-AIPP-Fsk labels both halves of P-glycoprotein, and labeling in the amino-terminal half can be localized to residues 291-359, which span proposed transmembrane regions 5 and 6 . KB-V1 membranes photolabeled with {125I}6-AIPP-Fsk and {125I}iodoarylazidoprazosin were digested with either Staphylococcus aureus V8 protease or chymotrypsin and had similar digestion patterns, suggesting that the two drugs label the same sites on P-glycoprotein.

Zh Mikrobiol Epidemiol Immunobiol, 1994 Aug-Sep, Suppl 1, 28 - 32
{The biological significance of the anticomplement activity of bacteria}; Brudastov IuA et al.; The occurrence and the average levels of anticomplementary activity among the representatives of bacterial species as Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, mainly reflecting differences between their ecovariants, were established . In connection with the fact that some of E . coli uropathogenic strains with high anticomplementary activity were found to be capable of inhibiting the activity of C5 in a hemolytic system and with the presence of correlation between the property under study and complement resistance, the existence of common mechanisms for the realization of these properties was supposed . Extracellular products of S . aureus were shown to be capable of inhibiting the functional activity and opsonic action of C3 component of the complement, which was related to the presence of anticomplementary activity in these bacteria . Experimental staphylococcal infection in mice revealed the predominant accumulation of bacteria possessing this property . Clinico-bacteriological studies demonstrated that the anticomplementary activity of E . coli contributed to the increased risk of postoperative complications in cholecystitis and to the prolongation of bacteriuria in chronic pyelonephritis.

J Chemother, 1994 Aug, 6(4), 219 - 25
Epidemiology of Staphylococcus aureus bacteremia in Denmark; Espersen F et al.; The number of Staphylococcus aureus bacteremia cases has increased annually in Denmark during the latest decades . This increase has occurred among older patients with hospital-acquired bacteremia . Methicillin-resistance, which earlier was a property of strains of the 83A phage-complex, has nearly disappeared, while the frequency of penicillin-resistance has increased . Today both the phage-type pattern and antibiotic resistance are nearly similar for strains from hospital-acquired and community-acquired cases . In hospitals the frequency of bacteremia after postoperative wound infections has decreased, while cases associated with intravascular catheters has increased, and these infections are now the most common cause of hospital-acquired S . aureus bacteremia . Endocarditis is most commonly found in community-acquired cases without an identified primary focus in patients between 21-50 years . Also hematogenous osteomyelitis is most common in community-acquired cases, but these infections have changed to having a high predilection for the vertebral column, and the prevalence of chronic cases has decreased.

J Antimicrob Chemother, 1994 Aug, 34(2), 261 - 5
In-vitro activity of new sulphanilil fluoroquinolones against Staphylococcus aureus; Allemandi DA et al.; Two new quinolones, NSFQ-104 and NSFQ-105, derivatives of ciprofloxacin and norfloxacin with a 4-(4-aminophenylsulphonyl)-1-piperazinyl at position 7 showed better in-vitro activity against strains of methicillin-susceptible and methicillin-resistant Staphylococcus aureus than ciprofloxacin or norfloxacin . Their in-vitro activity was enhanced at pH 5.5.

J R Coll Surg Edinb, 1994 Aug, 39(4), 214 - 7
A review of burns treated over a one-year period at Mpilo Central Hospital, Zimbabwe; Muguti GI et al.; A retrospective study is presented of 297 patients (57% male and 43% female) treated for burns at Mpilo Central Hospital, Bulawayo, Zimbabwe in 1991 . Of the patients, 73% were in the paediatric age group, 60% of whom were under 5 years of age . Most burns occurred at home (90%), with the majority occurring in the kitchen (61%) . The commonest burning agent was hot water (55%) . Open-fire burns accounted for 18% of cases . The surface area of burns ranged from 1% to 80% with a mean of 7% . The majority of patients (84%) had superficial thickness burns . Only 6% of the patients required active resuscitation with intravenous fluids . A total of 159 organisms were isolated on 111 pus swabs . The most commonly isolated organism was Staphylococcus aureus (66%) . Seven patients (five children and two adults) died, giving a mortality rate of 2% . Although it is desirable to have a purpose-built burns unit this study shows that satisfactory results can be achieved with limited facilities . In our practice, where 90% of burns occur in the home situation, an active public-education campaign would help to reduce the incidence of burns.

Chin Med J (Engl), 1994 Aug, 107(8), 570 - 3
Molecular epidemiologic study of burn wound infection caused by Staphylococcus aureus in children; Wu SX et al.; 115 children with burn injuries were admitted to the Children's Hospital of Chongqing Medical University . Of the 1554 samples, 276 strains of staphylococcus aureus were isolated from the burn wounds and other sites . The rate of burn wound infection caused by S . aureus was 25.2% (29/115) . The 54 epidemic strains of S . aureus all carried 1.6 and 1.9 Md plasmid DNAs belonging to phage type 618, and were resistant to at least 10 antimicrobial agents, including oxacillin, cephalothin and cephaloridine, but sensitive to tobramycin and amikacin . Identical plasmid profiles and phage types of isolated S . aureus indicated that a patient carrying a multi-resistant strain of S . aureus in his anterior nares caused an epidemic of S . aureus wound infection in 13 patients . S . aureus isolated from burn wounds of 8 cases were derived from the contaminated hands of their family members.

Zentralbl Hyg Umweltmed, 1994 Aug, 196(1), 52 - 69
{Nosocomial infections with the detection of Staphylococcus aureus in an average hospital--an 11-year analysis}; Heuck D et al.; Nosocomial infections over an 11 years period were monitored prospectively in a district hospital . A total of nosocomial infections among 162.197 patients discharged from 1980-1990 were analyzed . The incidence rate of nosocomial infections was 3.6% . The predominant isolates were Escherichia coli and Staphylococcus aureus (11%) . The highest rates of S . aureus infections were established in wound infections and skin and mucosal infections at surgery, gynaecology and paediatrics departments . The number of multiply resistant S . aureus strains has decreased and those of sensitive isolates has increased . 72% of 652 S . aureus isolates have been typed by phage typing and biotyping . Statistical analysis could not establish a significant relationship between phage patterns and nosocomial infections . The spectrum of phage patterns of these nosocomial strains was the same as those of the endogenous colonization of men . Nosocomial outbreaks could not been realized.

Bioorg Khim, 1994 Aug-Sep, 20(8-9), 842 - 56
{Study of the structural organization of OSCP--subunits of H+-ATPase of porcine heart mitochondria}; Grinkevich VA et al.; Unmodified and citraconilated OSCP of the pig heart mitochondrial H(+)-ATPase were hydrolysed by proteinase from Staphylococcus aureus V8 and trypsin, respectively . To purify the individual peptides, various types of HPLC and covalent chromatography on SH-Sepharose were used . By the automatic Edman method complete or partial amino acid sequences of the peptides obtained were determined, thus allowing for the reconstruction of the primary structure of pig OSCP . A linear antigenic determinant recognizable by A1 monoclonal antibody against bovine OSCP, was localized . Studies showed Gly43 residue (bovine OSCP) to be replaced by Ala43 (pig OSCP), which is responsible for a decrease of the affinity of the monoclonal antibody A1 to pig OSCP . Comparative analysis of primary structures of bovine and pig OSCP was carried out.

J Biochem (Tokyo), 1994 Aug, 116(2), 457 - 63
Evidence that the heparin-binding consensus sequence of vitronectin is recognized by Staphylococcus aureus; Liang OD et al.; Binding of heparin-binding form of vitronectin to Staphylococcus aureus was inhibited completely by heparin or by the same form of vitronectin . The binding was inhibited only to about 50% by the non-heparin-binding form of vitronectin, indicating an apparent involvement of the heparin-binding properties in the interaction between vitronectin and S . aureus . This was supported by experiments in which a synthetic peptide (Ala347-Arg361, comprising heparin-binding consensus sequences) was found to partly inhibit bacterial adherence to immobilized vitronectin . A bacterial cell surface protein could bind to the quinquedecapeptide, but not to the highly charged peptides consisting entirely of arginine or lysine, immobilized on microtiter plates and the binding could be competitively inhibited by an excess of soluble peptide . Direct binding of radiolabeled peptide to bacterial cells was also demonstrated, which was rapid, saturable, and pH-dependent . Furtherly a bacterial surface protein having molecular mass of 60 kDa was isolated by affinity chromatography on a quinquedecapeptide-HiTrap-NHS column . Our data suggest that the heparin-binding properties of vitronectin play a role in bacterial recognition.

Hybridoma, 1994 Aug, 13(4), 263 - 70
Characterization of neutralizing monoclonal antibodies directed against Staphylococcus aureus alpha-toxin; Heveker N et al.; A panel of neutralizing murine monoclonal antibodies (MAbs) against Staphylococcus aureus alpha-toxin has been established, using formaline-inactivated alpha-toxin as an immunogen . Five independent groups of neutralizing epitopes have been identified representing five functionally important structures in the toxin molecule . Because none of the antibodies binds to overlapping decapeptides representing the toxin sequence or to bromocyanogen cleavage products of alpha-toxin, they may all bind to conformational epitopes . Nevertheless, they all bind to monomeric alpha-toxin in a Western blot . Three of the antibodies bind to the toxin monomer in an enzyme-linked immunosorbent assay (ELISA) in the presence, but not in the absence, of detergent . These epitopes are not accessible in hexameric toxin; two of them may represent the contact sites of the toxin monomers upon hexamerization and one is related to a structurally important glycine-rich central hinge region . Two different antibodies bind to monomeric toxin in an ELISA in the presence and absence of detergent and their epitopes are present more than once on oligomeric toxin; they bind strongly to hexameric toxin in a Western blot . The binding properties of the antibodies to alpha-toxin in different assay systems are summarized in an epitope model, which describes the presence of neutralizing domains in the different conformational steps required for pore formation.

Antimicrob Agents Chemother, 1994 Aug, 38(8), 1808 - 12
Mechanistic studies and biological activity of bioxalomycin alpha 2, a novel antibiotic produced by Streptomyces viridodiastaticus subsp . "litoralis" LL-31F508; Singh MP et al.; The bioxalomycins, a novel complex of broad-spectrum antibiotics, were isolated from fermentations of Streptomyces viridodiastaticus subsp . "litoralis" LL-31F508 . Bioxalomycin alpha 2, the major component of this complex, exhibited antibacterial activity . The MICs ranged from < or = 0.002 to 0.008 micrograms/ml for gram-positive organisms and from 0.50 to 4 micrograms/ml for gram-negative organisms . Bioxalomycin alpha 2 was found to be bactericidal and to inhibit bacterial DNA synthesis preferentially . Bioxalomycin alpha 2 protected mice from a lethal challenge with Staphylococcus aureus Smith . The 50% effective dose of bioxalomycin alpha 2 administered orally was 10 times greater than that when the drug was given subcutaneously or intravenously . These data suggest a stability or bioavailability problem when the compound is administered orally.

Anal Chem, 1994 Aug 1, 66(15), 2465 - 70
Optical biosensor for monitoring microbial cells; Watts HJ et al.; The potential of a new optical biosensor, the resonant mirror, for detecting whole cells is demonstrated . Staphylococcus aureus (Cowan-1) cells, which express protein-A at their surface, were detected by binding to human immunoglobulin G (IgG) immobilized on an aminosilane-derivatized sensor surface at concentrations in the range 8 x 10(6)-8 x 10(7) cells/mL . A control S . aureus strain (Wood-46), which does not express protein-A, gave no significant response . Immobilization of the capture ligand on aminosilane surfaces with and without a hydrogel coating of carboxymethyl-dextran was compared . The greatest binding response was observed with non-dextran-coated surfaces . The sensitivity of the technique was increased a 1000-fold by using a human IgG-colloidal gold conjugate (30 nm) in a sandwich assay format . S . aureus (Cowan-1) cells were detected in spiked milk samples at cell concentrations from 4 x 10(3)-1.6 x 10(6) cells/mL using the sandwich assay.

J Immunol, 1994 Aug 1, 153(3), 1015 - 26
Interference with the binding of a naturally processed peptide to class II alters the immunodominance of T cell epitopes in vivo; Nikcevich KM et al.; T lymphocytes elicited in response to an immunizing Ag usually recognize only one or a few immunodominant peptides . The mechanisms governing this process are poorly understood . This study examines the consequences of peptide competition on immunodominance . Immunization of B10.A mice with the native Staphylococcus aureus nuclease protein primes T cells to the dominant 86-100 peptide presented in association with I-Ek class II molecules . To render the 86-100 peptide incapable of binding to the class II molecule, single amino acid substitutions were introduced in the native Staphylococcus aureus nuclease protein within a putative I-Ek class II binding motif . Introduction of residue changes at positions 89 and 91 in the protein prevents 86-100-specific T cell clone recognition of the protein in vitro . Competition studies demonstrate that substitutions at residues 89 or 91 decreased the I-Ek binding affinity of the 86-100 peptide . Immunization of B10.A mice with the L89F or Y91S mutant proteins does not prime T cells to the dominant 86-100 peptide; T cells are primed instead to I-Ek-restricted subdominant peptide(s) encompassed by the residues 111-135 . In vitro binding studies demonstrate that both the 111-130 and 116-135 synthetic peptides compete with a labeled I-Ek-binding peptide 20-fold less efficiently than the dominant 86-100 peptide, suggesting that these subdominant peptides may be of lower binding affinity than the dominant 86-100 peptide . These results support the hypothesis that dominance is dependent on peptide binding affinity for the appropriate class II molecule and the ability to compete with other peptides, derived from the same Ag, for class II binding.

Ned Tijdschr Geneeskd, 1994 Jul 30, 138(31), 1575 - 8
{Cerebellar abscess caused by an occipital dermal sinus in an infant}; Groen RJ et al.; A 7.5-month-old girl was admitted with the clinical signs and symptoms of raised intracranial pressure . This was caused by an obstructive hydrocephalus, due to cerebellar abscesses induced by an infected contiguous complete occipital dermal sinus . Staphylococcus aureus was cultured . The patient was treated by radical excision of the dermal sinus and the abscesses after initial external drainage of the lateral cerebral ventricle, followed by systemic antibiotic therapy . The different types of dermoid cysts of the posterior cranial fossa are described . Early detection of congenital dermal abnormalities along the craniospinal axis by routine examination of newborns is prognostically important . Computerised tomography or magnetic resonance imaging are the methods of choice for further investigation of suspect dermal lesions.

Ned Tijdschr Geneeskd, 1994 Jul 30, 138(31), 1571 - 4
{Methicillin-resistant Staphylococcus aureus in repatriates from the Faro airplane crash, 1992}; Stomph EC et al.; OBJECTIVE . To review the suitability of the guidelines of the Working Group Infection Prevention (WIP) and the Medical Inspectorate of Health with regard to possible methicillin-resistant Staphylococcus aureus (MRSA) contamination . SETTING . Medical Inspectorate of Health . DESIGN . Retrospective analysis . METHODS . For 95 repatriates from the Faro air crash, a questionnaire was filled out by medical microbiologists and attending specialists in Dutch hospitals (response rate: 100%) . Classification for MRSA risk category was done in accordance with the guidelines of the Medical Inspectorate of Health . Surveillance data of the National Institute of Public Health and Environmental Protection were consulted . RESULTS . Colonisation with MRSA occurred in 8 out of the 59 patients in the high-risk group (hospitalisation > 24 h in Portugal or invasive procedures) . No MRSA was found in the low-risk group (all other repatriates) . The MRSA-positive patients had burns at least . Infection was detected in three patients on admission, in five patients later (after 6-27 days of hospitalisation), after 1-9 negative cultures . Probably all patients had been contaminated with MRSA in Portugal . In two Dutch hospitals secondary contamination was observed . CONCLUSION . The WIP guidelines on MRSA control proved to be sufficient with exception of the inventory cultures and the isolation measures notably for patients with covered wounds . The MRSA risk classification of patients according to the guidelines of the Medical Inspectorate of Health proved adequate.

Ned Tijdschr Geneeskd, 1994 Jul 30, 138(31), 1565 - 7
{An outbreak of methicillin-resistant Staph aureus in a nursing home}; Peerbooms PG et al.; The introduction of methicillin-resistant Staphylococcus aureus (MRSA) in an Amsterdam nursing home (312 beds) by a patient discharged from a local hospital, the spread of this strain to other residents and staff of the nursing home and the actions taken to prevent this spread are described . During the period when this MRSA strain was circulating residents and staff members were checked periodically for presence of MRSA . In spite of all preventive measures as implemented in Dutch hospitals (including isolation of infected residents) additional residents (in a total of 8) and staff members acquired the MRSA strain . Relaxation of the precautions (no more isolation) and continued instruction of the members of the staff was attended by a decrease of further contamination . Four of 8 residents died while being colonised or infected . The course of this epidemic illustrates that limited precautions and proper training of staff in hygiene can restrict spread of MRSA in nursing homes.

Int J Cancer, 1994 Jul 15, 58(2), 248 - 53
Lymphotoxin-alpha/beta heterodimer is expressed on leukemic hairy cells and activated human B lymphocytes; Mapara MY et al.; The expression of human lymphotoxin (LT) alpha/beta cell-surface complex was studied in human B-cell lines as well as in normal and neoplastic human B lymphocytes . In the absence of TNF receptors, only the human hairy-cell leukemia (HCL)-derived cell line JOK-I revealed constitutive cell-surface expression of LT but not TNF-alpha . Immunoprecipitation experiments with anti-LT monoclonal antibody (MAb) 9B9 from cell-surface radioiodinated JOK-I cells revealed that a cell-surface lymphotoxin molecule (25 kDa) is expressed in association with a 33-kDa molecule . Enzymatic digestion with F/N-glycosidase and O-glycosidase showed that both proteins contained N-linked carbohydrate residues, whereas only the 25-kDa molecule contained O-linked sugar residues . Analysis of mRNA expression revealed specific transcripts of LT-alpha and LT-beta in JOK-I cells . Resting tonsillar B cells did not express cell-surface LT . However, LT-beta mRNA was observable in unstimulated tonsillar B cells, whereas LT-alpha mRNA, cell-surface LT and LT secretion could only be detected upon in vitro activation . Thus LT-beta and alpha appear to be sequentially expressed in human B cells . Neoplastic B cells from chronic lymphocytic leukemia (BCLL), being devoid of constitutive cell-surface LT expression, could be induced to express surface LT by in vitro stimulation with Staphylococcus aureus Cowan I (SAC) . Constitutive LT-beta transcripts, however, could also be detected in 4 out of 5 cases of BCLL . In contrast, human HCL cells displayed constitutive cell expression of lymphotoxin-alpha and beta . These findings demonstrate that cell-surface LT-alpha is expressed in association with LT-beta on activated normal B cells and neoplastic B cells representing an activated state.

Arch Intern Med, 1994 Jul 11, 154(13), 1505 - 8
Long-term efficacy of intranasal mupirocin ointment . A prospective cohort study of Staphylococcus aureus carriage; Doebbeling BN et al.; BACKGROUND: We investigated the long-term effect of a single 5-day application of intranasal mupirocin calcium ointment on Staphylococcus aureus nasal and hand colonization . The subjects were 68 healthy volunteers who were health care workers with stable S aureus nasal carriage and who had participated in a randomized, double-blind placebo-controlled clinical trial of intranasal mupirocin ointment . METHODS: A 1-year prospective cohort study of S aureus nasal carriers after treatment with active drug or placebo was performed . Cultures were obtained from all subjects 6 and 12 months after therapy . All subjects returned for the 6-month visit; 63 (93%) were examined at 1 year . The major outcome measure was the relative proportion of any S aureus cultured at either site at 6 and 12 months . The S aureus isolates were typed by restriction endonuclease analysis of plasmid DNA and by antibiotic susceptibility tests; the similarity of nasal and hand isolate "fingerprints" was compared . RESULTS: At 6 months, nasal carriage was 48% in the treatment group vs 72% in controls (relative risk, 0.68; 95% confidence interval, 0.45 to 1.02; P = .054); at 1 year, nasal carriage was 53% vs 76%, respectively (relative risk, 0.70; 95% confidence interval, 0.48 to 1.02; P = .056) . Hand carriage at 6 months was significantly reduced among mupirocin recipients relative to controls (15% and 48%; P = .04, adjusted for the baseline rate of hand carriage) . Thirty-six percent of treated subjects were recolonized in the nares with a new strain at 1 year, whereas 34% had reisolation of the original strain after initially negative posttherapy cultures . During the year of follow-up, hand carriage was observed at least once in two thirds of the subjects . Nearly all of the hand isolates (87%) exactly matched the subjects' coincident nasal plasmid fingerprint and antibiogram type . CONCLUSIONS: A single brief treatment course of intranasal mupirocin was effective in reducing nasal S aureus carriage for up to 1 year . When S aureus was recovered after nasal decolonization, the new isolate was as likely to represent colonization with a new strain as reisolation of the original strain . Staphylococcus aureus hand carriage was significantly decreased 6 months after therapy, further implicating the nares as the primary reservoir site for hand carriage.

Proc Natl Acad Sci U S A, 1994 Jul 5, 91(14), 6398 - 402
Mevalonate-mediated suppression of 3-hydroxy-3-methylglutaryl coenzyme A reductase function in alpha-toxin-perforated cells; Giron MD et al.; The regulation of mevalonic acid synthesis requires both nonsterol isopentenoid and sterol regulatory signal molecules . A primary target of this multivalent control process is the enzyme which catalyzes mevalonate synthesis: 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase (EC 1.1.1.34) . In this report Staphylococcus aureus alpha-toxin perforated Chinese hamster ovary cells were used to facilitate the identification of isopentenoidogenic reactions and metabolites required for mevalonate-mediated loss of HMG-CoA reductase activity . alpha-Toxin-perforated cells retained the capacity to decrease, upon demand, HMG-CoA reductase activity and protein in response to mevalonate or isopentenoid pyrophosphate esters . Also, it was deduced with highly specific metabolic inhibitors, that conversion of farnesyl 1-diphosphate to squalene was required for mevalonate-mediated suppression of reductase activity . Since squalene (2 microM) did not downregulate reductase activity, pre-squalene pyrophosphate or a derivative, or polyprenyl-1-pyrophosphate-generated inorganic pyrophosphate, or a combination of these metabolites are proposed as candidate regulatory nonsterol isopentenoid signal molecules.

J Biomed Mater Res, 1994 Jul, 28(7), 775 - 82
Lens-on-surface method for investigating adhesion of Staphylococcus aureus to solid surfaces incubated in blood plasma; Elwing H et al.; Adhesion of Staphylococcus aureus was investigated on flat silicon oxide surfaces that had been incubated in human plasma at different concentrations . Adhesion of bacteria did not occur at high incubation concentrations of plasma or when the surface had been incubated in egg albumin . However, significant adhesion was observed when plasma was diluted . With the use of antibody method, it was noted that the adhesion of the bacteria coincided with adsorbed fibrinogen, and possibly also with IgG . We also investigated the effect of "narrow space" on the adsorption of blood plasma and subsequent adhesion of S . aureus . In these experiments, blood plasma was incubated under a convex lens placed upside-down on the silicon oxide surface . This method creates a continuous gradient of space from the contact point of the lens and outward . After rinsing off the plasma and the lens, the surface was incubated with a suspension of S . aureus followed by quantification of the attached bacteria by means of optical methods . Adhesion of bacteria occurred in several circular zones that were easily detectable with the naked eye or by the means of simple optical methods . In addition, in these experiments, adhesion coincided with adsorbed fibrinogen or IgG at the surfaces . The increased bacterial adhesion to surfaces incubated in diluted plasma, or plasma incubated in narrow space, is a variant of the so-called "Vroman effect." With a model protein system consisting of fibrinogen and IgG and the corresponding antibodies, we demonstrate that "dilution" and "incubation in narrow space" are two phenomenologically similar methods.(ABSTRACT TRUNCATED AT 250 WORDS)

J Antibiot (Tokyo), 1994 Jul, 47(7), 821 - 32
Synthesis of 2''-amino-2''-deoxyarbekacin and its analogs having potent activity against methicillin-resistant Staphylococcus aureus; Kondo S et al.; Based on our studies on the enzymatic modifications of arbekacin by methicillin-resistant Staphylococcus aureus (MRSA), replacement of the 2''-hydroxyl group by an amino group in arbekacin was designed to synthesize derivatives that would be active against MRSA . 2''-Amino-2''-deoxyarbekacin and five analogs were synthesized starting from dibekacin . Among them, 2''-amino-2''-deoxyarbekacin and the 5-epiamino analog showed excellent antibacterial activities against not only MRSA but also Gram-negative bacteria including Pseudomonas, and lower toxicities than arbekacin.

Leukemia, 1994 Jul, 8(7), 1146 - 52
Leukemic cells from progressive B-CLL respond strongly to growth stimulation in vitro; Aguilar-Santelises M et al.; Isolated leukemic B cells from patients with B-chronic lymphocytic leukemia (B-CLL) were tested for proliferative response in vitro to Staphylococcus Aureus strain Cowan 1 (SAC), IL-2, and low molecular weight (MW) BCGF . Patients were classified according to clinical stage and progressiveness . Ten of eighteen cell populations from patients with progressive B-CLL responded in vitro with a stimulation index (SI) > 20 . Only 1/16 non-progressive patients had a proliferative but low response . Normal unfractionated tonsillar B cells responded to SAC and BCGF, whereas normal high buoyant density B cells were unresponsive . After 3 days of stimulation, responding B-CLL cells had multiplied and the B cells expressed CD5, CD19, and weakly CD21 . No cells in the responding cultures exhibited CD3 or the EBV nuclear antigen EBNA-1 . Cell maturation, measured as IgM secretion, was found in some, but not in all responding B-CLL cultures . Thus, B-CLL cells from patients with progressive disease have the capacity to respond to signaling through surface Ig receptors and to certain T-cell factors which was not the case for B-CLL cells from non-progressive patients . The pattern of in vitro response may be related to disease progression, reflecting a dependency of normal immunoregulatory mechanisms and/or a dysregulation of the growth control in the leukemic cells.

Eur J Biochem, 1994 Jul 1, 223(1), 303 - 8
The thermostability of natural variants of bacterial plasminogen-activator staphylokinase; Gase A et al.; Three natural variants (wild-type staphylokinase, {R36G, R43H}staphylokinase, and {G34S, R36G, R43H}staphylokinase) of the bacterial plasminogen-activator staphylokinase, a 136-amino-acid protein secreted by certain Staphylococcus aureus strains, have been characterized . These variants differ at amino acid positions 34, 36 and 43 only, and have a very similar plasminogen-activating capacity and conformation in solution, as revealed by fluorescence spectroscopy, dynamic light scattering and circular dichroism . However, the thermostability of these variants is significantly different . At 70 degrees C and 0.5 mg protein/ml, irreversible inactivation occurred with apparent half-life (t1/2) values 0.54 +/- 0.13, 0.81 +/- 0.20 and 3.7 +/- 0.7 h (mean +/- SEM) for wild-type staphylokinase, {R36G, R43H}staphylokinase, and {G34S, R36G, R43H}staphylokinase, respectively, with corresponding values at 0.08 mg/ml of 5.3 +/- 1.4 h and 11 +/- 2.0 h for wild-type staphylokinase and {R36G, R43H}staphylokinase, respectively . Dynamic light-scattering measurements indicated that inactivation was associated with protein aggregation, which precluded accurate determination of transition temperatures and enthalpies of unfolding . 0.08-0.34 mg/ml {G34S, R36G, R43H}staphylokinase, however, did not aggregate at 70 degrees C but underwent unfolding as revealed by a 20% increase in the Stokes' radius and a 30% decrease in circular dichroism . The unfolding was reversible upon cooling and was associated with full recovery of functional activity . Thus, these natural variants of staphylokinase have a different sensitivity to thermal inactivation, that is mediated by reversible unfolding of the protein and concentration-dependent irreversible aggregation . {G34S, R36G, R43H}staphylokinase, the most resistant natural variant, has a stability approaching the minimal requirements for pasteurization, which would facilitate its development for clinical use.

Am J Respir Cell Mol Biol, 1994 Jul, 11(1), 95 - 102
Characteristics of the inhibition of NADPH oxidase activation in neutrophils by apocynin, a methoxy-substituted catechol; Stolk J et al.; Phagocytes are able to generate reactive oxygen species by an activatable NADPH oxidase system . We investigated the inhibition of NADPH oxidase activation by a methoxy-substituted catechol, apocynin . Oxygen uptake by neutrophils incubated with 300 microM apocynin was completely inhibited at 7 min after addition of serum-treated zymosan (STZ), with a lagtime of inhibition of 2 to 3 min . The lagtime of effect of apocynin in neutrophils relatively deficient of myeloperoxidase was about 50% longer when compared with normal cells . Inhibition of the STZ-induced respiratory burst by apocynin was also observed in human eosinophils but not in human alveolar macrophages . Immunoblots of neutrophil membranes, isolated at 2 and 7 min after STZ stimulation of neutrophils, demonstrated translocation of the cytosolic oxidase components p47-phox and p67-phox to the membrane fraction . Translocation at 7 min after STZ stimulation was markedly reduced when the neutrophils had been incubated with 300 microM apocynin, but translocation was normal after 2 min of stimulation . These properties suggest that apocynin is an intracellular inhibitor of the assembly of NADPH oxidase in neutrophils and eosinophils and that apocynin requires conversion by peroxidases to exert its inhibitory effect . The capacity of neutrophils for intracellular killing of Staphylococcus aureus was not affected by apocynin . The potential therapeutic value of apocynin was demonstrated in vitro by its ability to protect secretory leukocyte proteinase inhibitor from oxidative inactivation by neutrophils.

Zh Mikrobiol Epidemiol Immunobiol, 1994 Jul-Aug, (4), 59 - 63
{Europium-labelled Staphylococcus aureus protein A as a reagent for determining specific antibodies}; Guzaeva TV et al.; In this work the conditions of labeling protein A with europium ions were studied and the conjugates obtained in this study were compared with traditional peroxidase conjugates currently used in immunochemistry . The conjugates of protein A with Eu3+ chelate were obtained with the use of cyclic dianhydride of diethylenetriaminepentaacetic acid (DADETPA) . Conjugation methods with the use of DADETPA was shown to permit obtaining high-quality conjugates with europium chelates . Europium-labeled protein A ensured the sensitivity of the determination of adsorbed IgG at a level of 2 ng/ml and the dynamic analytical range within 3-1,000 ng/ml, which essentially exceeded similar characteristics of peroxidase conjugates with protein A . Europium-labeled protein A was used for the detection of antibodies to Francisella tularensis in the sera of humans immunized against tularemia . The sensitivity of this assay exceeded that of the enzyme immunoassay 10- to 40-fold . A conclusion was made on the possibility of using europium labelled protein A for the determination of specific antibodies to F.tularensis . This preparation may be useful in the determination of specific antibodies in low-immune sera.

Zh Mikrobiol Epidemiol Immunobiol, 1994 Jul-Aug, (4), 49 - 51
{The use of solid-phase immunoenzyme analysis for the diagnosis of a suppurative infection of the maxillofacial area caused by Bacteroides melaninogenicus and Staphylococcus aureus}; Iastrebova NE et al.; In 186 patients with odontogenic infection the level of antibodies to B.melaninogenicus polysaccharide antigen, teichoic acid and S.aureus polysaccharide was determined in the enzyme-linked immunosorbent assay (ELISA) . The positive values of ELISA indices were found to coincide with the data of bacteriological study in more than 90% of cases . The role of B.melaninogenicus and S.aureus associations in the development of purulent infections was confirmed with the use of ELISA in 34.2% of the patients for S.aureus and in 50% of the patients for B.melaninogenicus and bacteriologically in 31.4% and 46.9% of the patients respectively.

Antimicrob Agents Chemother, 1994 Jul, 38(7), 1664 - 7
Mupirocin resistance among consecutive isolates of oxacillin-resistant and borderline oxacillin-resistant Staphylococcus aureus at a university hospital; Layton MC et al.; Mupirocin resistance was determined in consecutive oxacillin-resistant and borderline oxacillin-resistant Staphylococcus aureus clinical isolates collected over 14 months at a university hospital during 1991 and 1992 . Twenty of 86 (23%) oxacillin-resistant and borderline oxacillin-resistant S . aureus isolates were mupirocin resistant; 80% were high-level resistant . Prior mupirocin use was a significant risk factor (relative risk, 6.08; 95% confidence interval, 3.7 to 9.99) . Seven of 20 resistant isolates were distinct strains, as determined by pulsed-field gel electrophoresis typing . Two instances of clonal dissemination of a single strain occurred, but several other distinct mupirocin-resistant strains were documented . Mupirocin resistance was unexpectedly common among these isolates.

Antimicrob Agents Chemother, 1994 Jul, 38(7), 1649 - 50
Production of A and C variants of staphylococcal beta-lactamase by methicillin-resistant strains of Staphylococcus aureus; Norris SR et al.; Most methicillin-resistant Staphylococcus aureus (MRSA) strains produce beta-lactamase . To determine whether this enzyme(s) is identical to one or more of the four beta-lactamases produced by methicillin-susceptible strains, the beta-lactamases of 50 MRSA isolates were typed by using substrate profile analysis . Forty type A, no type B, ten type C, and no type D beta-lactamase-producing strains were identified . The beta-lactamase inhibitor sulbactam reduced the MICs of beta-lactamase-labile antibiotics, including ampicillin, penicillin G, and cefazolin, for type A and type C MRSA strains.

Antimicrob Agents Chemother, 1994 Jul, 38(7), 1515 - 8
Minocycline versus vancomycin for treatment of experimental endocarditis caused by oxacillin-resistant Staphylococcus aureus; Nicolau DP et al.; The purpose of this study was to determine the penetration of minocycline and vancomycin into cardiac vegetations and to determine their efficacy in a rabbit model of endocarditis caused by oxacillin-resistant Staphylococcus aureus . Animals were randomized into three groups: control (no antibiotic), minocycline (6 mg/kg given intravenously every 8 h), and vancomycin (50 mg/kg given intravenously every 8 h) . Penetration of the antibiotics into aortic valve vegetations was determined by using the tissue/serum area under the concentration-time curve ratio . The reductions in the bacterial density of the vegetations caused by both vancomycin (4.8 +/- 1.2 CFU/g) and minocycline (5.3 +/- 1.6 CFU/g) were significantly different from that of controls (8.7 +/- 1.8 CFU/g) . Although the penetration of minocycline was twice that of vancomycin, they were equally effective in reducing the bacterial density of the vegetations, since the concentrations of both agents in tissue remained above their MICs for oxacillin-resistant S . aureus . For organisms for which the MICs are higher, however, these penetration differences may result in treatment differences.

Protein Eng, 1994 Jul, 7(7), 925 - 31
Increased solubility of trimethoprim-resistant type S1 DHFR from Staphylococcus aureus in Escherichia coli cells overproducing the chaperonins GroEL and GroES; Dale GE et al.; The production of the trimethoprim-resistant type S1 dihydrofolate reductase (DHFR) from Staphylococcus aureus in Escherichia coli cells overproducing the chaperonins GroEL and GroES is described . The simultaneous overproduction of the chaperonins with DHFR results in an increased solubility of the enzyme . We compare the time course of production of active type S1 DHFR by measuring enzyme activity in cells overproducing or not overproducing the chaperonins . Although co-overproduction of the chaperonins reduces the total production level of type S1 DHFR, the amount of soluble and active DHFR is increased several-fold in comparison with cells producing only DHFR . Thus, the higher concentrations of GroES and GroEL in cells overproducing the chaperonins partially protect DHFR from aggregation, resulting in higher concentrations of soluble and active DHFR in the cell . Furthermore, we also demonstrate that the chaperonins can improve in vitro refolding yields of type S1 DHFR . These results suggest that it is possible to purify suitable amounts of trimethoprim-resistant type S1 DHFR for X-ray crystallographic studies.

Indian J Exp Biol, 1994 Jul, 32(7), 492 - 4
Effect of trace metals on production of exoprotein and beta-lactamase by Staphylococcus aureus; Bhat KG et al.; A minimum concentration of 0.01 mM of Mg2+ was essential for any growth of S . aureus . Of the trace metals, Mg2+ and Ca2+ were highly stimulatory for growth . All trace metals except Ni2+ in small concentrations were stimulatory for growth and exoprotein production . Amounts of exoprotein and beta-lactamase produced relative to the number of viable cells were maximum during the period of suboptimal proliferation of bacteria . Trace metals did not significantly influence the amount of beta-lactamase produced relative to exoprotein.

Leuk Lymphoma, 1994 Jul, 14(3-4), 311 - 8
Proliferative response of B chronic lymphocytic leukemia lymphocytes stimulated with IL2 and soluble CD23; Brizard A et al.; The in vitro proliferative response of purified B-chronic lymphocytic leukemia (B-CLL) lymphocytes cultured in the presence of soluble CD23 (sCD23) with or without IL2 was compared to the responses induced by phorbol 12-myristate 13-acetate (PMA), Staphylococcus aureus strain Cowan I (SAC), IL1, IL2, IL4, IL6 and the combination of IL2 and interferon (IFN) alpha or IFN gamma . As expected, B-CLL lymphocytes proliferated with PMA, SAC and IL2 with a clear enhancement of the IL2-induced response by IFN alpha or IFN gamma . They failed to proliferate in response to sCD23, IL1, IL4 or IL6 alone nor to the combinations of sCD23 and any of the 3 latter cytokines . However, sCD23 significantly increased the proliferation of B-CLL cells induced by IL2, suggesting a protective effect of sCD23 on apoptosis . Serum levels of sCD23 and CD23 membrane expression were high in every patient which is compatible with the hypothesis of an autocrine or paracrine activation loop . Detectable CD23 expression was lost in all cultures except for that stimulated by PMA . Only supernatants of PMA-stimulated cultures contained high sCD23 levels.

Intern Med, 1994 Jul, 33(7), 454 - 5
Rhabdomyolysis associated with bacteremic pneumonia due to Staphylococcus aureus; Bando T et al.; A case of rhabdomyolysis associated with bacteremic pneumonia due to Staphylococcus aureus is reported . A 58-year-old man was admitted because of severe left lobar pneumonia, and presented myoglobinuria on admission . The serum creatine phosphokinase level was transiently elevated and myoglobinuria were disclosed . Sputum and venous blood cultures grew Staphylococcus aureus . Rhabdomyolysis has rarely been reported in cases of bacterial infection, especially those due to Staphylococcus aureus . This case might be an extremely rare case of rhabdomyolysis complicating Staphylococcus aureus bacteremia following severe left upper lobar pneumonia.

Ann Plast Surg, 1994 Jul, 33(1), 68 - 71
Unilateral galactocele following augmentation mammoplasty; Deloach ED et al.; Development of unilateral galactoceles following breast augmentation is reported in 2 young females . Both galactoceles were drained and cultured . In 1 patient the implant was removed and a delayed reinsertion was undertaken . In the second patient the implant was replaced at the time of the drainage procedure . Culture and sensitivity in 1 patient showed no growth and in the second patient revealed Staphylococcus aureus . Although the cause is unknown, galactocele formation may be due to manipulation of breast tissue during surgery . The use of oral contraceptives may also play a role in this process . Hormonal suppression of lactation and removal of the implants may be indicated in these patients . Consideration should be also given to the use of systemic antibiotics directed toward skin pathogens.

Rev Clin Esp, 1994 Jul, 194(7), 535 - 9
{Study of 164 episodes of infectious endocarditis in drug addicts: comparison of HIV positive and negative patients}; Valencia ME et al.; OBJECTIVE . Intravenous drug-addicts (IVDA) are the most important risk group for acquiring HIV and developing infective endocarditis (IE) in Spain . The clinical and evolutive features of a large series of IE in HIV-infected IVDA are reported and compared with those observed in a smaller group of patients without HIV infection . METHODS . A retrospective analysis was made of 164 episodes of IE from 136 patients diagnosed in a Service of Infectious Diseases in Madrid (1986-1992) . IE was defined according to the modified Von Reyn criteria and only the probable and definite IE episodes were evaluated . RESULTS . A total of 143 IE episodes occurred in 117 IVDA infected with HIV (group A) and 21 episodes in 19 HIV-negative patients (group B) . Most of seropositive patients were asymptomatic carriers of HIV (54%) and only 22% had AIDS . The IE onset was acute for 97% patients in group A and 81% in group B, with fever and respiratory symptoms as main complaints . Chest X-ray was normal in 19% of cases in group A and in 28% in group B . Septic embolisms were observed in 56% and 41% of patients in group A and B, respectively . The vegetation originated mainly on the tricuspid valve and Staphylococcus aureus was recovered from most blood-cultures . The mortality rate was similar in both groups, 6% and 5% in groups A and B, respectively . CONCLUSIONS . Most IVDA with IE were HIV-positive patients in this series . IE is usually reported in the early stages of HIV infection and apparently its presence has no influence on the clinical course of IE.

Naunyn Schmiedebergs Arch Pharmacol, 1994 Jul, 350(1), 96 - 9
The inhibitory effects of allopurinol on the production and cytotoxicity of tumor necrosis factor; Olah T et al.; Allopurinol, a xanthine oxidase inhibitor, impaired the cytotoxic effect of human recombinant tumor necrosis factor (TNF) against WEHI cells . Actinomycin D abolished the inhibition of cytotoxicity by allopurinol . Allopurinol also exerted an inhibitory effect on the production of TNF by human mononuclear cells stimulated by either heat-killed Staphylococcus aureus or E . coli lipopolysaccharide . It is suggested that allopurinol inhibits TNF cytotoxicity by decreasing the level of oxygen free radicals generated (among other mechanisms) by the action of xanthine oxidase . Whatever the mechanism, the fact that allopurinol counteracts the toxicity of TNF can help towards an understanding of the complex nature of TNF toxicity.

Jpn J Antibiot, 1994 Jul, 47(7), 869 - 72
{Transferability of biapenem (L-627) to cerebrospinal fluid in rabbits with meningitis caused by Staphylococcus aureus}; Haruta T et al.; The transferability of biapenem (L-627) to cerebrospinal fluid (CSF) was studied employing rabbits with experimental meningitis caused by Staphylococcus aureus . The mean plasma concentration was 192 +/- 12.8 micrograms/ml at 15 minutes after intravenous administration of the drug at a dose level of 100 mg/kg . The mean concentration in CSF was maximum at 45 minutes after administration at 11.4 +/- 2.19 micrograms/ml . Pharmacokinetic parameters calculated from these values were as follows, Cmax (CSF/plasma): 5.96%; AUC (CSF/plasma): 8.15% between 15 and 60 minutes, 12.1% between 15 and 120 minutes and 15.0% between 15 and 180 minutes; T1/2 for L-627 in CSF: 152 minutes; T1/2 (CSF/plasma): 3.34 . In comparison with those of other beta-lactam antibiotics that were obtained in the same way, the transferability of L-627 was intermediate, and in consideration of the antimicrobial potency against the main pathogens of meningitis, it appears worthwhile of running clinical trials for this drug.

J Clin Microbiol, 1994 Jul, 32(7), 1830 - 2
Rapid detection of methicillin-resistant Staphylococcus aureus by Crystal MRSA ID System; Qadri SM et al.; A commercially available method for the rapid detection of methicillin-resistant Staphylococcus aureus (BBL Crystal MRSA ID System) was evaluated and compared with conventional methods . All 52 isolates of methicillin-susceptible and 142 isolates of intrinsic methicillin-resistant S . aureus were correctly identified in 4 h by the test method, whereas correct identification took 11 to 24 h by conventional methods . The test is simple, rapid, and easy to perform and the results are easy to interpret.

J Exp Med, 1994 Jul 1, 180(1), 95 - 109
Leukocytosis and resistance to septic shock in intercellular adhesion molecule 1-deficient mice; Xu H et al.; Intercellular adhesion molecule 1 (ICAM-1) is one of three immunoglobulin superfamily members that bind to the integrins lymphocyte function associated 1 (LFA-1) and Mac-1 on leukocytes . We have generated mice that are genetically and functionally deficient in ICAM-1 . These mice have elevated numbers of circulating neutrophils and lymphocytes, as well as diminished allogeneic T cell responses and delayed type hypersensitivity . Mutant mice are resistant to lethal effects of high doses of endotoxin (lipopolysaccharide {LPS}), and this correlates with a significant decrease in neutrophil infiltration in the liver . Production of inflammatory cytokines such as tumor necrosis factor alpha or interleukin 1 is normal in ICAM-1-deficient mice, and thus protection appears to be related to a diminution in critical leukocyte-endothelial interactions . After sensitization with D-galactosamine (D-Gal), ICAM-1-deficient mice are resistant to the lethal effect of low doses of exotoxin (Staphylococcus aureus enterotoxin B {SEB}), which has been shown to mediate its toxic effects via the activation of specific T cells . In this model, ICAM-1-mediated protection against SEB lethality correlates with a decrease in the systemic release of inflammatory cytokines, as well as with prevention of extensive hepatocyte necrosis and hemorrhage . ICAM-1-deficient mice sensitized with D-Gal, however, are not protected from lethality when challenged with low doses of endotoxin (LPS) . These studies show that the different contribution of ICAM-1 in the activation of either T cells or macrophages is decisive for the fatal outcome of the shock in these two models . This work suggests that anti-ICAM-1 therapy may be beneficial in both gram-positive and -negative septic shock, either by reducing T cell activation or by diminishing neutrophil infiltration.

Antibiot Khimioter, 1994 Jul, 39(7), 39 - 41
{Antibiotic prophylaxis and reconstructive operations in patients with burn sequelae}; Popov SV et al.; The results of the clinicolaboratory investigation of the efficacy of antibiotic prophylaxis in 215 patients subjected to reconstructive operations for postburn scars are presented . The biological investigation of the postburn scars revealed the presence of microflora in about half of the patients . The microflora was mainly represented by Staphylococcus aureus and S . epidermidis . The level of the dissemination was not high . Ulcerated scars had higher levels of the bacterial contamination . The pharmacokinetic study of cefazolin, carbenicillin, aztreonam and gentamicin showed that the release of the first three drugs to the scar tissues was good . The clinical observations demonstrated that purulent inflammatory complications developed in patients with risk factors (ulceration of the scar, necrotic foci in the wound, involvement of the oral cavity and nasal passages to the operation), not treated with the antibiotics or treated with the drugs not inhibiting the microflora of the operation wounds (gentamicin, aztreonam) . The results permitted to recommend the use of the antibiotic prophylaxis in reconstructive operations only in risk patients with burn consequences . With this purpose it is advisable to use antibiotics satisfactorily penetrating to the tissues of the postburn scars and active against the microflora vegetating in them (cefazolin and carbenicillin).

Zentralbl Veterinarmed B, 1994 Jul, 41(5), 328 - 35
Virulence for mice of Staphylococcus aureus strains from bovine mastitis related to colonial morphology and serological types in serum-soft agar; Calvinho LF et al.; Six Staphylococcus aureus strains isolated from bovine mastitic milk representing the typical growth types in soft agar media were injected into mice via the intraperitoneal route . Strains showing diffuse colony morphology (DCM) in serum-soft agar (SSA) as a permanent characteristic which reacted against anti-capsular sera types A, B, and D were virulent for mice . A strain showing DCM in SSA that reacted only against anti-capsular serum D behaved as a compact-colony-morphology-type strain in the peritoneal cavity of the mouse . Diffuse-type colony morphology and presence of capsular antigens type A, B, and D correlated with increased virulence for mice, but a capsule could not be demonstrated.

Rev Argent Microbiol, 1994 Jul-Sep, 26(3), 124 - 32
{Recovery of Staphylococcus aureus after acid injury in milk products}; Assis EM et al.; The growth behavior of Staphylococcus aureus in fresh Cheese (Minas and Muzzarella) during their shelf-life was studied . The possible injury of this microorganism caused by the increasing acidity was also investigated . Raw milk was inoculated with 10(6) cells/ml (S . aureus FRIA-100) and the cheese production was performed according to normal procedures . Minas and muzzarella cheese were stored at 7 degrees C for 40 and 60 days, respectively . At 2-3 days intervals, the following analysis were performed: acidity, pH, S . aureus counting using agar Baird Parker by the traditional methods and by the method recommended by the American Public Health Association to evaluate the reparation of injured cells . We had a secure indication of the presence of injured S . aureus when acidity was in the range of 0.7 to 0.8% expressed in lactic acid and when the cycle was 1.3 log higher than the traditional one.

Eur J Clin Microbiol Infect Dis, 1994 Jul, 13(7), 559 - 64
Prospective evaluation of a two-week course of intravenous antibiotics in intravenous drug addicts with infective endocarditis . Grupo de Estudio de Enfermedades Infecciosas de la Provincia de Cádiz; Torres-Tortosa M et al.; In a prospective study, a two-week course of antibiotics (cloxacillin 2 g/4 h plus amikacin 7.5 mg/kg/12 h) was evaluated in the therapy of right-sided infective endocarditis in intravenous drug users (IVDU) . All IVDU admitted to hospital during the study period who fulfilled the strict criteria for diagnosis of infective endocarditis were analysed . A subgroup of patients with right-sided endocarditis caused by Staphylococcus aureus who had a good prognosis were selected as being eligible for the two-week course of treatment . In a total of 139 episodes of infective endocarditis in IVDU, 72 (51.8%) cases were eligible for the two-week treatment . Of this group, 67 were cured, 4 needed prolongation of treatment to cure the infection and 1 died in hospital of respiratory distress syndrome on day 10 of treatment . In patients not eligible for the two-week treatment, the mortality was higher (24.2% versus 0.7%; p = 0.00015) . Drug toxicity in the treated group was low . It can be concluded that administration of cloxacillin and amikacin parenterally for 14 consecutive days was successful in the therapy of right-sided endocarditis in IVDU.

Rev Latinoam Microbiol, 1994 Jul-Sep, 36(3), 191 - 6
{Investigation of the presence of Staphylococcus aureus in cooked ham using 3 methods (preliminary study)}; Uscanga-Prieto I et al.; Fifty-one cooked ham samples were analized by three methods for investigation of Staphylococcus aureus; Modified Van Doorne and American Public Health Association; The Official Mexican Method . The first two are enrichment methods and in all three a comparison between Baird Parker agar and salt milk agar was done . The modified Van Doorne technique was the best for isolation of S . aureus from cooked ham . In Baird Parker agar it was possible to demonstrate the presence of S . aureus in all positive samples . The study shows the importance of using Baird Parker broth as an enrichment medium for the isolation of S . aureus from products in which a thermal treatment and addition of salts as sodium chloride and nitrites, inhibit the growth of this microorganism.

Rev Latinoam Microbiol, 1994 Jul-Sep, 36(3), 171 - 6
{Characterization of a transpositional mutant of Staphylococcus aureus underproducing exoproteins}; Giraudo AT et al.; A pleiotropic transpositional mutant derived from Staphylococcus aureus strain RC46, from bovine origin, was isolated and characterized . This mutant showed decreased production of several exoproteins such as alpha- and beta-hemolysins, DNase, coagulase and extracellular protein A . The production of cell-bound protein A, proteases and delta-hemolysin was not affected . The pleiotrophy of this mutant, designated sae (for S . aureus exoprotein expression), is due to a single insertion of transposon Tn917, as indicated by Southern blot hybridization . The sae mutant showed decreased virulence since its LD50 determined by intraperitoneal injection in mice was 32 times higher than that of the parental strain RC46.

Adv Ren Replace Ther, 1994 Jul, 1(2), 167 - 75
Managing Staphylococcus aureus catheter infection in continuous ambulatory peritoneal dialysis patients; Holley JL et al.; A case of Staphylococcus aureus catheter infection in a patient on peritoneal dialysis is presented and discussed by nephrologists, a social worker, a nurse specializing in the care of peritoneal dialysis patients, and the patient involved . The focus of the multidisciplinary case discussion concerns the management of S aureus catheter infections, including catheter removal, psychosocial issues and the patient's response to the need for catheter removal, the risk factors and prevention of S aureus catheter infections in peritoneal dialysis patients, and exit site care practices.

Adv Ren Replace Ther, 1994 Jul, 1(2), 107 - 18
Causes of hemodialysis access failure; Albers FJ; Complications of hemodialysis access remain significant problems in the population receiving renal replacement therapy . The causes of access loss must be recognized before appropriate interventions can be designed . The native primary arteriovenous fistula is the access of choice because of its good survival characteristics and low rate of complications . Unfortunately, a substantial number of patients have vasculature insufficient to create and maintain this access . Once a primary fistula is established, thrombosis is the leading cause of failure: its causes can be divided into early (less than 6 weeks) and late complications . In patients unable to have fistulas, arteriovenous conduits of expanded polytetrafluoroethylene (ePTFE) are now the prostheses of choice . Again thrombosis is the leading cause of this access loss but there is also a substantial rate of failure from infection, pseudoaneurysms, perigraft hematomas, and simple attrition of the prosthesis . Thrombosis of ePTFE grafts is usually associated with anatomic stenosis at the venous anastomosis, within the graft itself, or in the central venous system . Graft thromboses cannot always be attributed to anatomic lesions: in these circumstances, thrombosis has been attributed to low-flow states . It is possible that the normal balance between endothelial procoagulant and anticoagulant forces are disrupted within the arteriovenous conduit . Recurrent venipuncture, hemodialysis therapy itself, or pathophysiological forces inherent in the access may all favor hemostasis . Infection also causes significant hemodialysis access morbidity . The causative organism is usually Staphylococcus aureus but several other pathogens have been reported . Ideally, all prostheses should be removed when infected, but this approach must be tempered by the reality of limited hemodialysis access sites . There is no consensus as to the best therapeutic approach to access infection.

J Clin Invest, 1994 Jul, 94(1), 97 - 104
Interleukin 10 induces B lymphocytes from IgA-deficient patients to secrete IgA; Briere F et al.; We have previously shown that human B lymphocytes cultured in the CD40 system, composed of an anti-CD40 mAb presented by a CD32-transfected fibroblastic cell line, proliferate but do not secrete antibodies . However, the addition of particles of Staphylococcus aureus Cowan (SAC) induces B cell differentiation even in the absence of exogenous cytokines (CD40/SAC system) . Additionally, B lymphocytes cultured in the CD40 system in the presence of human IL-10, produce IgM, IgG, and IgA, and Ig levels are further increased by SAC . Here, we have studied the capacity of peripheral blood lymphocytes from patients with IgA deficiency (IgA-D) to secrete Igs, particularly IgA after CD40 triggering . Peripheral blood mononuclear cells (PBMNC) from IgA-D patients cultured in the CD40/SAC system produced IgM and IgG, but not IgA . The addition of IL-10 to the cultures, enhanced the production of IgM and IgG and most strikingly induced the production of high amounts of IgA . The addition of IL-10 to PBMNC from IgA-D patients activated through CD40 alone resulted in the production of IgA . Thus, SAC and anti-CD40 mAb stimulate B cells to differentiate into cells secreting IgG and IgM whereas IL-10 plays a central role in inducing B cells from IgA-D patients to differentiate into IgA secreting cells.

Cell Immunol, 1994 Jul, 156(2), 493 - 507
Inhibition of human B lymphocyte cell cycle progression and differentiation by rapamycin; Aagaard-Tillery KM et al.; In this study, we have analyzed the effects of the immunosuppressive agent rapamycin on the activation of highly purified normal human B lymphocytes . When the polyclonal activators Staphylococcus aureus (SA) and soluble CD40 ligand (CD40L) were used to stimulate B cells, rapamycin inhibited both interleukin 2 (IL2)-dependent and -independent proliferation, as well as IL2-dependent differentiation into antibody-secreting cells . Cell cycle analysis indicated that rapamycin inhibited the progression of SA+IL2-stimulated B cells past the mid-G1 phase of the cell cycle . To begin to identify rapamycin-sensitive signaling events essential for B cell activation, we examined the effects of rapamycin on p34cdc2 and p33cdk2 kinase activities . SA+IL2 stimulation induced the activation of both cyclin-dependent kinases . Of interest, rapamycin abrogated the activation of both p34cdc2 and p33cdk2 . Our results indicate therefore that rapamycin inhibits a number of SA- and CD40L-inducible events that may be necessary for both entry into S phase and for permitting subsequent B cell differentiation . These studies emphasize the utility of this drug as a tool to begin to dissect the activation pathways utilized by human B cells, as well as to provide implications for the therapeutic use of rapamycin in vivo.

Ugeskr Laeger, 1994 Jun 27, 156(26), 3871 - 5
{Septic arthritis}; Andersen K et al.; Septic arthritis (SA) is reviewed in the English literature with reference to registration of localisation of the infected joints, the bacteriological cause, predisposing factors, clinical information and laboratory data related to SA . Seventy-eight publications were available concerning 975 patients with 1086 infected joints caused by 1032 bacteria . Large joints such as the knee and hip were most frequently, however, infected, in recent years, SA in small axial joints has been reported with increased frequency . Gram-positive cocci and especially Staphylococcus aureus are most frequently the cause of infection . High temperatures, elevated sedimentation rate and leucocytosis are often noticed in SA, as is a warm, tender, swollen joint with effusion and painful movements . Investigation of the aspirated fluid from the joint for bacteria and number of leucocytes as well as CT-scanning or scintigraphy are the most important diagnostic tools . With rapid and correct antibiotic treatment the prognosis for good or excellent function of the joint is reported to be from 27-90%, while the mortality is reported as being 7-23%.

J Biol Chem, 1994 Jun 17, 269(24), 16821 - 8
Protein kinase C in yeast . Characteristics of the Saccharomyces cerevisiae PKC1 gene product; Antonsson B et al.; The Saccharomyces cerevisiae PKC1 gene encodes a homolog of mammalian protein kinase C (Levin, D . E., Fields, F.O., Kunisawa, R., Bishop, J.M., and Thorner, J . (1990) Cell 62, 213-224) . A protein of 150 kDa is recognized by a polyclonal antiserum raised against a trpE-Pkc1 fusion protein . In subcellular fractionations, Pkc1p associates with the 100,000 x g particulate fraction . This association is resistant to extraction with high salt concentrations, alkali buffer, or nonionic detergents, suggesting that Pkc1p may be associated with a large protein complex . Pkc1p modified at its COOH terminus with two repeats of the Staphylococcus aureus protein A IgG-binding fragment (ZZ sequence tag) was able to fully restore the growth defects of a pkc1ts strain at restrictive temperature . ZZ-tagged Pkc1p was partially purified by chromatography on DEAE-Sepharose, followed by IgG-Sepharose . In vitro, Pkc1p phosphorylates the pseudosubstrate peptide and myelin basic protein, but not histones . Replacing an isoleucine with an arginine 2 amino acids COOH-terminal of the acceptor serine in the substrate peptide resulted in a 10-fold decrease of Km . Pkc1p activity was independent of cofactors such as phospholipids, diacylglycerol, and Ca2+, known to activate several mammalian protein kinase C isoenzymes, making it a rather distantly related member of the protein kinase C superfamily.

FEMS Microbiol Lett, 1994 Jun 15, 119(3), 263 - 9
Cloning and expression of the penicillinase from a borderline methicillin-susceptible Staphylococcus aureus strain in Escherichia coli; Massidda O et al.; The blaZ gene contained in a single 17.2-kb beta-lactamase plasmid from a borderline methicillin-susceptible Staphylococcus aureus strain (a53) has been cloned in Escherichia coli . A Bluescript II derivative in which the ampicillin resistance gene has been replaced with the chloramphenicol resistance gene was used as a multi-copy vector . One ampicillin-resistant colony was detected among 31 chloramphenicol-resistant transformants selected . This E . coli clone harbored a recombinant plasmid (pAH12) containing two different staphylococcal HindIII inserts (7.0 and 5.3 kb), of which only the former hybridized with a blaZ probe . The clone showed an ampicillin MIC of > 1024 micrograms ml-1, independently of the inoculum size used, and produced large amounts of beta-lactamase, which hydrolyzed nitrocefin and penicillin G but not methicillin of the beta-lactamase substrate, padac . In contrast, S . aureus a53 hydrolyzed all four substrates . The fact that high levels of staphylococcal penicillinase are unable to cause methicillin hydrolysis confirms that penicillinase hyperproduction is unlikely to be the true mechanism responsible for the borderline phenotype . These results also suggest that the two different beta-lactamases (penicillinase and methicillinase) associated with borderline S . aureus strains have a different genetic origin.

Eur J Biochem, 1994 Jun 15, 222(3), 919 - 25
Purification of a bone sialoprotein-binding protein from Staphylococcus aureus; Yacoub A et al.; Bone sialoprotein (BSP) is selectively bound by Staphylococcus aureus cells isolated from patients suffering from infections of bone and joint tissues {Ryden C., Maxe, I., Franzen, A., Ljungh, A., Heinegard, D . & Rubin, K . (1987) Lancet II, 515} . We now report on the purification of a cell-wall protein from Staphylococcus aureus, strain O24, that possesses affinity for bone sialoprotein . Staphylococcal cell-wall components with capacity to inhibit binding of 125I-labeled BSP to staphylococcal cells were solubilized with LiCl (1.0 M, pH 5.0) . Preparative SDS/PAGE and protein-overlay experiments revealed that inhibitory activity present in LiCl extracts resided in a fraction of polypeptides with M(r) 75,000-110,000 . Staphylococcal proteins solubilized with LiCl were chromatographed on a Mono-Q anion-exchange column . Inhibitory activity was eluted at 0.6-0.8 M NaCl and could be further purified by affinity chromatography on BSP-Sepharose . Elution of the affinity matrix with 0.1 M glycine, pH 3.0, specifically eluted inhibitory activity . Analysis by SDS/PAGE revealed a single M(r) 97,000 polypeptide in the eluate . The purified M(r) 97,000 protein bound BSP in protein-overlay experiments . LiCl extracts from S . aureus, strain E514 or Staphylococcus epidermidis, strain 7686, both lacking the capacity to bind BSP did not contain the M9r) 97,000 protein . Our data demonstrate the presence of a S . aureus cell-surface BSP-binding protein . This protein could be involved in bacterial tropism in osteomyelitis.

Biochemistry, 1994 Jun 14, 33(23), 7477 - 84
Identification of a putative membrane-inserted segment in the alpha-toxin of Staphylococcus aureus; Ward RJ et al.; To gain a fuller understanding of the regions of the Staphylococcus aureus alpha-toxin important in pore formation, we have used Forster dipole-dipole energy transfer to demonstrate that a central glycine-rich region of alpha-toxin (the so-called "hinge" region) inserts deeply into the bilayer on association of toxin with liposomes . Mutant alpha-toxins with unique cysteine (C) residues at positions 69 and 130 {Palmer, M., et al . (1993) J . Biol . Chem . 268, 11959) were reacted with the C-specific fluorophore acrylodan, which acted as an energy donor . The chosen acceptor was N-(7-nitrobenz-2-oxa-13- diazol-4-yl)-1,2-bis(hexadecanoyl)-sn-glycero-3-phosphoethanolamin e (NBD-PE) . Measurement of the degree of donor quenching with increasing NBD-PE in the inner bilayer leaflet enables the distance of closest approach between donor and acceptor to be estimated . For toxin labeled with acrylodan at position 130 (in the hinge region), this distance is approximately 5 +/- 2 A, showing that the probe is close to the inner surface of the liposomes . A second probe labeled at position 69 (in the N-terminal domain) shows negligible energy transfer, indicating a distance of closest approach > 40 A . This implies that this N-terminal region remains "outside" the liposome . We propose a model in which the central region of the alpha-toxin inserts into the membrane and possibly participates in forming the wall of the pore.

Biochemistry, 1994 Jun 14, 33(23), 7294 - 9
Arsenate reductase of Staphylococcus aureus plasmid pI258; Ji G et al.; Arsenate reductase encoded by Staphylococcus aureus arsenic-resistance plasmid pI258 was overproduced in Escherichia coli and purified . The purified enzyme reduced radioactive arsenate to arsenite when coupled to thioredoxin, thioredoxin reductase, and NADPH . NADPH oxidation coupled to arsenate reduction also required thioredoxin and thioredoxin reductase . Glutaredoxin and reduced glutathione did not stimulate arsenate reduction . NADPH oxidation showed Michaelis-Menten kinetics with a Km of 1 microM AsO4(3-) and an apparent Vmax of 200 nmol/min per mg of protein . At high substrate concentration (above 1 mM AsO4(3-), a secondary rise in the reaction rate was observed, with a Km of 2 mM and an apparent Vmax of 450 nmol/min per mg of protein . This secondary rise also occurred upon addition of phosphate or nitrate (which were not substrates for the enzyme) . Arsenite (the product of the enzyme), tellurite, and antimonite {Sb(III)} were inhibitors . Selenate (but not selenite or sulfate) was a substrate for reductase-dependent NADPH oxidation, with an apparent Km of 13 mM SeO4(2-) . Arsenate reductase was purified as a monomer of 14.5 kDa, consistent with the DNA sequence . Electrospray mass spectrometry showed two molecular masses of 14,810.5 and 14,436.0 Da, suggesting that 70% of the purified protein lacked the N-terminal three amino acids; HPLC coupled to electrospray mass spectroscopy of protease digest products confirmed this conclusion and verified the entire amino acid sequence.

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