J Biol Chem, 1994 Jun 3, 269(22), 15563 - 70 Interaction of N-terminal fragments of fibronectin with synthetic and recombinant D motifs from its binding protein on Staphylococcus aureus studied using fluorescence anisotropy; Huff S et al.; The N-terminal 29-kDa fragment of fibronectin (Fn29K) contains five type I "finger" modules . It binds to heparin, fibrin, and bacteria and is involved in fibronectin (Fn) matrix assembly . Binding to Staphylococcus aureus involves a cell wall-associated protein that contains approximately three repeats of a 38-residue D motif (Signas, C., Raucci, G., Jonsson, K., Lindgren, P.-E., Anantharamaiah, G.M., Hook, M., and Lindberg, M . (1989) Proc . Natl . Acad . Sci . U.S.A . 86, 699-703) . Synthetic peptides representing D1, D2, and D3, when labeled with fluorescein isothiocyanate (FITC), exhibited increases in fluorescence anisotropy upon addition of Fn29K but not other Fn fragments . The response could be reversed by titration with unlabeled peptides to yield inhibition constants that agreed with the dissociation constants obtained by fitting the initial response . Values of Kd ranged between 2 and 12 microM, with D3 having the highest affinity . Specificity of D3 for Fn29K was further illustrated by the fact that its C-terminal half (D3b, Lys801 to Lys821), when immobilized, selectively adsorbed Fn29K from a thermolysin digest of fibronectin . The binding site in Fn was further localized within Fn29K by analyzing smaller proteolytic or recombinant subfragments . Those containing fingers, F3-5 and F4-5, were purified on D3b-Sepharose and bound FITC-D3b with Kd values of 4-6 microM . Subfragments containing pairs of fingers 1-2, 2-3, or single fingers 1, 4, or 5 were inactive . Whole D1-3, expressed in Escherichia coli and labeled with fluorescein, bound 1.9 mol/mol of Fn29K with Kd = 1.5 nM . F4-5 and F2-3 bound with respective Kd values of 0.35 and 4.4 microM . These and other results indicate that binding of the individual D region peptides is mediated through their C-terminal halves, primarily to fingers 4 and 5 of fibronectin . The possible basis of the much higher affinity of D1-3 is discussed. J Bacteriol, 1994 Jun, 176(12), 3445 - 54 Transcriptional regulation by TrsN of conjugative transfer genes on staphylococcal plasmid pGO1; Sharma VK et al.; The major conjugative transfer gene cluster of staphylococcal plasmid pGO1 (trs) consists of 13 open reading frames (trsA to trsM) transcribed from one DNA strand and a single 189-bp open reading frame (trsN) within the first 348 bp of trs that is transcribed divergently . Promoter regions for trsN and trsA partially overlap . TrsN, a 7,181-Da protein, was purified as a fusion to glutathione S-transferase and found to have DNA-binding activity . Increasing concentrations of the fusion protein progressively retarded the gel migration of PCR-generated DNA fragments containing predicted promoters 5' to trsL, trsA, and trsN . The target sequences contained areas of identity, including regions of dyad symmetry, that were protected in DNase I footprinting studies . The binding of TrsN to its trsL target was required for this target DNA to be stably introduced into Staphylococcus aureus on a high-copy-number vector . Provision of excess TrsN from this high-copy-number vector in S . aureus decreased beta-galactosidase activity from a trsL-lacZ transcriptional fusion and decreased pGO1 conjugation frequency . Conversely, both transcription and conjugation increased in the presence of excess trsL target . We propose that TrsN negatively regulates the transcription of genes essential for conjugative transfer by binding to regions 5' to their translational start sites. Am J Gastroenterol, 1994 Jun, 89(6), 924 - 7 Bacterial endocarditis in patients with chronic liver disease; McCashland TM et al.; OBJECTIVE: Although patients with cirrhosis have an increased susceptibility for bacterial infections, endocarditis complicating cirrhosis has been reported only infrequently . In this study, our objective was to determine whether, bacterial endocarditis is, in fact, a complicating factor in cirrhosis . METHODS: We retrospectively studied all cases of bacterial endocarditis that occurred over the last 15 yr in patients with known cirrhosis . RESULTS: Ten patients (three males, seven females) were identified, whose mean age was 55 yr (range 29-65 yr) . Bacterial organisms included Staphylococcus aureus, coagulase-positive (eight patients), Peptostreptococcus (one patient), and Enterococcus (one patient) . Underlying liver disease consisted of alcoholism (five patients), autoimmune chronic active hepatitis (two), cryptogenic cirrhosis (two), and primary biliary cirrhosis (one) . Distribution of heart valves affected were mitral valve (six), aorta (two), and there were two involving both mitral and aortic valves . Echocardiograms revealed vegetation in 50% of the patients . Laboratory studies were markedly abnormal, with mean values of albumin 2.4 mg/dl, creatinine 2.5 mg/dl, BUN 76.5 mg/dl, and total bilirubin 8.2 mg/dl . Potential associated sources of infection were upper gastrointestinal bleeding (four), pneumonia (two), and one each of spontaneous bacterial peritonitis, hip replacement, heart catheterization, and abdominal abscess . The outcome was poor, with death in eight of 10 patients . CONCLUSIONS: Bacterial endocarditis may complicate cirrhosis, may be more frequent in females, typically involves the mitral valve, and probably is due to Staphylococcus aureus. Am J Gastroenterol, 1994 Jun, 89(6), 915 - 23 The contribution of vitamin K2 (menaquinones) produced by the intestinal microflora to human nutritional requirements for vitamin K; Conly JM et al.; BACKGROUND: Coagulopathy manifest by elevation of the prothrombin time (PT) in patients receiving broad spectrum antimicrobials indirectly suggests a role for intestinal microflora synthesized menaquinone (MK) in the maintenance of normal coagulation . Nonetheless, no direct evidence is available to support this contention . OBJECTIVE: Our objective was therefore to provide evidence that bacterially produced MK may be absorbed by the distal small bowel of humans . METHODS: Using a cell harvester, Staphylococcus aureus (ATCC 29213) was grown in 12-L batches, harvested, and extracted by high performance liquid chromatography (HPLC) to obtain 8 mg of pure MK . Four normal volunteers were placed on a diet severely restricted in vitamin K1 (median 32-40 U/day), and were given warfarin to maintain an International Normalized Ratio of approximately 2.0 . On the 10th day of warfarin administration, naso-ileal intubation was performed and 1.5 mg of MK was delivered into the ileum . PT, factor VII, II and serum vitamin K1 levels were monitored throughout the study . RESULTS: Mean serum vitamin K1 levels were reduced to 30% of the pre-diet value at the time of MK administration . Within 24 h of ileal MK administration, there was a significant (p < 0.05) increase in the factor VII level of 0.28 +/- 0.10 U/ml (mean +/- SEM) and a significant decrease of 2.5 (+/- 0.1) s in the PT, whereas in the control phase (during which no MK was administered), there were no significant changes in the PT or factor VII at corresponding time intervals . CONCLUSION: These data provide direct evidence for the absorption of vitamin K2 from the distal small bowel, supporting a definite role for bacterially synthesized vitamin K2 in contributing to the human nutritional requirements of this vitamin. Infect Immun, 1994 Jun, 62(6), 2478 - 82 Corneal virulence of Staphylococcus aureus: roles of alpha-toxin and protein A in pathogenesis; Callegan MC et al.; Staphylococcus aureus produces a variety of proteins, including alpha-toxin and protein A, that could contribute to corneal tissue damage during keratitis . We examined corneal infections produced by intrastromal injection of four S . aureus strains--three isogenic mutants, one lacking alpha-toxin (Hly- Spa+), one lacking protein A (Hly+ Spa-), and one lacking both alpha-toxin and protein A (Hly- Spa-), and the wild type (Hly+ Spa+)--in a rabbit model of experimental keratitis . Rabbit corneas were injected intrastromally with 100 CFU of one of the four strains, and the eyes were examined by slit lamp biomicroscopy over a 25-h period . Corneal homogenates were used for determination of CFU and neutrophil myeloperoxidase activity at 5-h intervals . All strains had the same logarithmic growth curve from 0 to 10 h postinfection, after which CFU remained constant at 10(7) CFU per cornea . By 15 h postinfection, slit lamp examination scores were significantly higher for eyes infected with Hly+ strains than for Hly(-)-infected eyes . At this time, distinct epithelial erosions were seen in Hly(+)-infected eyes but not in Hly(-)-infected eyes . Myeloperoxidase activity was significantly greater for Hly(+)-infected corneas than for Hly(-)-infected corneas at both 20 and 25 h postinfection . Spa(+)- and Spa(-)-infected eyes showed no differences in slit lamp examination scores or myeloperoxidase activities . These results suggest that alpha-toxin, but not protein A, is a major virulence factor in staphylococcal keratitis, mediating the destruction of corneal tissue in eyes infected with this bacterial pathogen. Infect Immun, 1994 Jun, 62(6), 2334 - 44 Immunopathological features of rat Staphylococcus aureus arthritis; Bremell T et al.; Staphylococcus aureus is the most common bacterial species found in nongonococcal bacterial arthritis in humans . We present the first description, to our knowledge, of an outbreak of spontaneous staphylococcal arthritis in a rat colony . In a group of 10 rats, 9 displayed arthritis . Clinically, the most obvious findings were arthritis of one or both hindpaws and malaise . Bacteriophage typing showed the common phage type 85 in isolates recovered from the joints, blood, and bedding of rats and from the nose and cheeks of one person from the staff of the animal facility . The S . aureus strain proved to produce staphylococcal enterotoxin A and exhibited strong binding to collagen types I and II and bone sialoprotein, which are potentially important virulence factors . When the recovered S . aureus strain was injected intravenously into healthy rats, severe septic arthritis was induced in almost all of the animals . The arthritic lesions were characterized by infiltration of phagocytic cells and T lymphocytes into the synovium . Many of the synovial cells strongly expressed major histocompatibility complex class II molecules . Increased levels of interleukin 6 in serum as well as a prominent polyclonal B-cell activation were noted throughout the disease course . Pretreatment of S . aureus-injected rats in vivo with an antibody to the alpha beta T-cell receptor significantly decreased the severity of the arthritis . Our results indicate that alpha beta + T lymphocytes contribute to an erosive and persistent course of S . aureus arthritis. Acta Paediatr Jpn, 1994 Jun, 36(3), 244 - 9 A flow cytometric method for measurement of hydrogen peroxide generation by pediatric polymorphonuclear leukocytes stimulated by Staphylococcus aureus and Escherichia coli; Ihara T et al.; The generation of hydrogen peroxide (H2O2), one of the reactive oxygen species, by polymorphonuclear leukocytes (PMN) stimulated by Staphylococcus aureus and Escherichia coli was studied in infants by cytofluorography . After heparinized whole blood was incubated with bacteria for 60 min, generated H2O2 was measured . The positive rate of H2O2 generation of PMN and mean fluorescent intensity of positive PMN stimulated by S . aureus and E . coli were significantly reduced in infants aged < 1 year and H2O2 generation increased with advancing age . In 10-15 year old children, the level of generated H2O2 reached adult levels . When sera from 1 year old children were added to separated PMN from healthy adults, H2O2 generation was reduced . In contrast, H2O2 generation by PMN from 1 year old children was increased by the addition of adult sera . These results suggest that the ability to generate H2O2 in response to S . aureus and E . coli is lower in infants and that such reduced activity may be related to the susceptibility of such infants to S . aureus and E . coli infections. Unfallchirurgie, 1994 Jun, 20(3), 174 - 8 {Delayed abscesses after unstable pelvic ring fractures of the lumbar spine . Case report from the viewpoint of the expert witness}; Dorges J et al.; As a result of medical certificates concerning traumatology and internal medicine the course of disease of a 51-year-old man after serious contusion with pelvic ring dislocation and lumbar vertebral fractures with extensive retroperitoneal abscesses is reported whereby the diagnosis and surgery of this complication has followed only five years after the accident . The significance of nosocomial infections by staphylococcus aureus in connection with closed injuries is pointed out; qualification and quality of the medical certificates are also referred to. Clin Infect Dis, 1994 Jun, 18(6), 992 - 4 Cervical vertebral osteomyelitis presenting as a retropharyngeal abscess; Faidas A et al.; Retropharyngeal abscess as a manifestation of nontuberculous cervical vertebral osteomyelitis in adults has been described rarely in the literature . We report a case of cervical vertebral osteomyelitis that was caused by Staphylococcus aureus and that presented as a retropharyngeal abscess in a 66-year-old man . In addition, we review related cases. Clin Infect Dis, 1994 Jun, 18(6), 942 - 5 Activation of the cytokine network in a patient with AIDS and the recalcitrant erythematous desquamating disorder; Dondorp AM et al.; An AIDS patient with the recalcitrant erythematous desquamating (RED) disorder, a presumed variant of the Staphylococcus aureus toxic shock syndrome that is characterized by relapses, skin involvement, and variable multiple-organ involvement, is described . A strain of S . aureus producing toxic shock syndrome toxin 1 was isolated from the patient, but no antibodies to this exotoxin were detectable before and during the most severe episode of the RED disorder . Levels of cytokines were measured during episodes and at other times . Tumor necrosis factor and interleukin 6 were detectable only during the most severe exacerbation of the disorder; this finding suggests a pathogenetic role for these cytokines. Clin Infect Dis, 1994 Jun, 18(6), 863 - 7 Osteomyelitis due to Bacillus cereus in an adolescent: case report and review; Schricker ME et al.; Non-anthracis Bacillus species associated with clinical infections are usually dismissed as contaminants or nonpathogens . As opportunists, however, Bacillus organisms can cause significant systemic infections including bacteremia, endophthalmitis, and pneumonia . Osteomyelitis with non-anthracis Bacillus organisms has been described in adults, although to our knowledge it has been described only once in a child . We report a case of chronic osteomyelitis due to Staphylococcus aureus and superinfection with Bacillus cereus in a 13-year-old adolescent . A Bacillus isolate should be considered a true pathogen in children with chronic osteomyelitis who have a poor clinical response to antistaphylococcal therapy. J Dairy Sci, 1994 Jun, 77(6), 1503 - 8 Effect of polyphosphate and sodium chloride on the growth of Listeria monocytogenes and Staphylococcus aureus in ultra-high temperature milk; Rajkowski KT et al.; With UHT-sterilized milk as a model system, combinations of polyphosphate (.5 and 1.0%) and NaCl (.5 and 4.5%) were studied to determine their effects on the growth kinetics of Listeria monocytogenes Scott A and Staphylococcus aureus 196E . The milk was inoculated with 10(3) to 10(4) cfu/ml of either L . monocytogenes or S . aureus and incubated under aerobic conditions at 12, 19, 28, or 37 degrees C . The addition of polyphosphate did not significantly inhibit the growth of either microbe at the temperatures studied, but the addition of NaCl or a combination of salts significantly inhibited growth . The addition of .5 or 1.0% polyphosphate alone to dairy products is not likely to affect substantially the growth of S . aureus or L . monocytogenes. Clin Nephrol, 1994 Jun, 41(6), 370 - 6 Effect of sodium fusidate and ofloxacin on Staphylococcus aureus colonization and infection in patients on continuous ambulatory peritoneal dialysis; Sesso R et al.; The effectiveness of sodium fusidate and ofloxacin to eliminate nasal and catheter exit-site Staphylococcus aureus colonization and to prevent infections was compared in 31 patients on continuous ambulatory peritoneal dialysis (CAPD) . In a prospective randomized study, 9 patients were treated with topical 2% sodium fusidate ointment applied in the anterior nares and in the pericatheter skin twice daily for 5 days; 9 subjects received oral ofloxacin 200 mg taken every 48 hours for 5 days and 13 subjects were in the control group . Treatment courses were repeated at one-month intervals . Mean duration of follow-up was 7.8 months (242 patients-month) . Follow-up samples from the nares and the catheter exit-site were obtained every month from all participants to determine the presence of S . aureus . Development of S . aureus exit-site infection and peritonitis were assessed . During the study, S . aureus was recovered from 45%, 59% and 52% of the samples from the nares and/or exit-site in the sodium fusidate, ofloxacin and control groups, respectively (p = 0.13) . S . aureus grew less frequently (p < 0.01) in samples from the exit-site in the sodium fusidate than in the other two groups . Eradication of nasal colonization (two negative cultures within one month) was observed in 43%, 40% and 33% of the cases in the sodium fusidate, ofloxacin and control groups, respectively (p > 0.50) . The corresponding figures for exit-site eradication were 43%, 33% and 11%, respectively (p = 0.34) . Twenty-four S . aureus-associated infection episodes (12 of exit-site and 12 of peritonitis) were diagnosed in 16 of the 31 patients.(ABSTRACT TRUNCATED AT 250 WORDS) Planta Med, 1994 Jun, 60(3), 218 - 21 Antimicrobial agents from Heterotheca inuloides; Kubo I et al.; By bioassay directed fractionation, four sesquiterpenoids 1-4 were isolated as antimicrobial agents from the dried flowers of Heterotheca inuloides, a Mexican medicinal plant locally known as "arnica" . 7-Hydroxy-3,4-dihydrocadalin (3) and 7-hydroxycadalin (4) exhibited potent antibacterial activity against Gram-positive bacteria with minimum inhibitory concentrations (MICs) ranging from 6.25 to 12.5 micrograms/ml . Notably, 7-hydroxy-3,4-dihydrocadalin showed bactericidal activity against methicillin-resistant Staphylococcus aureus (MRSA) with a minimum bactericidal concentration (MBC) of 12.5 micrograms/ml. Poult Sci, 1994 Jun, 73(6), 904 - 15 The role of environment and management on leg abnormalities in meat-type fowl; Hester PY; Continuous light with 1 h of darkness at midnight is a common photoperiod for raising meat-type fowl . The logic behind the 1 h of darkness, usually provided between midnight and 0100 h, is to acclimate the birds to darkness in the event of a power failure . Increasing evidence from several research laboratories indicates that lighting regimens other than continuous light, such as intermittent or step-up lighting, can lower the incidence of leg abnormalities in meat-type fowl . Some evidence suggests that increased exercise contributes in part to the reduction in lameness due to lighting . Management can influence the incidence of leg and foot problems via effects on rate of gain, flooring systems, and litter moisture . Rapid weight gains have been correlated with tibial dyschondroplasia (TD), although more evidence is needed to determine whether other types of leg deformities, such as long bone distortion, are related to rapid growth rates . Using low intensity X-ray imaging (hand-held lixiscope), live breeders with TD lesions can be identified and the incidence of TD can be reduced in breeder flocks through genetic selection . In addition, turkeys can be selected for wider shanks to improve walking ability . Slippery surfaces should be avoided to prevent spraddled legs . With the exception of TD, broilers reared in cages have more leg deformities than floor-reared birds . Dry litter conditions can help prevent foot pad dermatitis caused by Staphylococcus aureus and other bacteria. Jpn J Antibiot, 1994 Jun, 47(6), 826 - 36 {Treatment with arbekacin of surgical infections by resistant strains of Staphylococcus aureus . Arbekacin Study Group}; Morimoto K et al.; The frequency of infection by methicillin-resistant Staphylococcus aureus (MRSA) is high in Japan and control of such strains is urgently needed . Arbekacin (ABK), a semisynthetic aminoglycoside, has potent activity against S . aureus, including resistant strains, and against Gram-negative bacteria as well . For this reason, in surgical infections (which are often caused by more than one bacterium), this drug might be particularly effective . We calculated the MIC and the decrease in the MIC when cultures of 59 resistant strains of S . aureus isolated in our wards at Osaka City University Hospital, contained arbekacin in the medium . We also used the drug to treat 12 infections caused by resistant strains of S . aureus . The MICs of vancomycin had a single peak at 0.5 microgram/ml, and those for ABK had double peaks at 0.5 and 4.0 micrograms/ml . The effect of arbekacin in lowering the MIC of minocycline (MINO) was slight because of the low MIC of MINO . Effects on fosfomycin (FOM), ampicillin, clavulanic acid/ticarcillin, cefotiam, cefuzonam, flomoxef, and imipenem/cilastatin were strong; the peaks were lowered by 1/2(7)-1/2(11) . When 1.0 micrograms/ml ABK was present in the medium, the efficacy of FOM was increased enough that, by prediction from the pharmacokinetics of FOM (blood level when given at the usual dose), all but one (2%) of the 47 resistant strains would be eradicated clinically . If 2.0 micrograms/ml ABK were in the medium, all strain would be eradicated, by our calculations . We treated 11 infections and one colonization by resistant strains of S . aureus with ABK and evaluated the response in these cases of infection . Four infections were treated with FOM as well . The clinical efficacy was good in four infections (three patients), fair in four, and poor in three, for an efficacy rate of 36% . All presumed causative bacteria were eradicated in two (18%) of the 11 infections and S . aureus strains were eradicated in three (27%) of the 11 infections . No symptoms of side effects were reported, but blood urea nitrogen and creatinine rose in a 72-year-old woman with duodenal perforation and peritonitis . The MIC levels of ABK were satisfactory, but clinical efficacy for staphylococcal infections caused by resistant strains was unsatisfactory. Jpn J Antibiot, 1994 Jun, 47(6), 820 - 5 {Efficacy of arbekacin, a new aminoglycoside antibiotic, in surgical patients with MRSA infections}; Ishikawa S et al.; The clinical efficacy of a new aminoglycoside antibiotic, arbekacin (ABK), was studied in surgical patients who had been infected with methicillin-resistant Staphylococcus aureus (MRSA) . Six cases of pneumonia, 2 of wound infections and 2 of intra-abdominal infections were treated by ABK alone or ABK together with beta-lactam antibiotics such as imipenem/cilastatin or cefotiam . The overall clinical efficacies against these MRSA infections were excellent in one case, good in 6 and poor in 3 . In six cases treated by ABK alone, good clinical responses were obtained in 4 cases . Among 4 cases that received combination therapy with ABK, good responses were obtained in 3 cases . No adverse reactions were found in ABK monotherapy or in combined therapy . These data suggested that ABK is an effective antibiotic on surgical infections caused by MRSA. Jpn J Antibiot, 1994 Jun, 47(6), 813 - 9 {MRSA infections in surgery}; Fujita K et al.; Susceptibilities to antibiotics were determined in 36 strains of methicillin-resistant Staphylococcus aureus (MRSA) isolated from clinical specimens from 1990 to 1992 . Rates of resistance to arbekacin and minocycline were 31% and 53%, respectively . However, all MRSA isolates were susceptible to vancomycin . MRSA was found in 12 out of 35 cases . Three infections caused by MRSA included enterocolitis (3), abscess (5), pneumonia (1), cholangitis (1), peritonitis (1) and catheter related sepsis (1) . In two cases patients died with bacteremia within two years after the onset of MRSA infections. Jpn J Antibiot, 1994 Jun, 47(6), 804 - 12 {Clinical effect of the combined therapy of arbekacin and imipenem/cilastatin against methicillin-resistant Staphylococcus aureus}; Hashimoto A et al.; We evaluated the efficacy of combined therapy of arbekacin (ABK) and imipenem/cilastatin (IPM/CS) against infections by methicillin-resistant Staphylococcus aureus (MRSA) . The MICs of ampicillin, cefmetazole, cefotiam, cefuzonam, flomoxef, fosfomycin, ofloxacin, minocycline, ABK and IPM/CS against clinically isolated strains of MRSA were examined . Almost all strains of MRSA were resistant to these antibiotics except ABK . Furthermore, combination of ABK and IPM/CS showed smaller MICs than that of ABK or IPM/CS alone . All fractional inhibitory concentration indices (FIC indices) of ABK plus IPM/CS were lower than 0.75 . The efficacy rate of combined therapy of ABK and IPM/CS in 22 patients with MRSA infections (15 patients with pneumonia, 3 patients with chronic bronchitis, 2 patients with sepsis, a patient with subcutaneous abscess and a patient with DPB) was 68% . And no patients had adverse reactions . Six (27%) of 22 strains of MRSA were eradicated . Significant correlations were found between bacteriological effect and severity of disease, and between serum albumin level and clinical effect. Jpn J Antibiot, 1994 Jun, 47(6), 741 - 50 {Clinical efficacy of arbekacin on MRSA pneumonia}; Kawashima T et al.; Arbekacin sulfate (ABK) was administered by intravenous drip to pneumonia patients infected with methicillin-resistant Staphylococcus aureus (MRSA), and the efficacy and the safety were objectively evaluated by the executive committee . The daily dose was determined in principle as 150-200 mg, two times a day, 30-90 minutes drip infusion, and the dose was to be changed at each special occasion . Combined therapy with other antibiotics was scheduled in severe cases at a decision of the physician in charge . Data of 18 cases were accumulated . The efficacy could be evaluated for 12 cases (4 cases with ABK alone, and 8 cases with combined therapy), and the safety was evaluated for 18 cases . The clinical efficacy was: excellent, 1; good, 4; fair, 5; and poor, 2 . The efficacy rate was 41.7% . The bacteriological effect was: eradicated, 2 (16.7%); decreased, 2; and no change, 8 . There found no side effects except 3 cases of abnormal laboratory data, two abnormal renal functions(11.1%) and one abnormal hepatic function (5.5%) . In one of the renal disorders, decreased dose of ABK improved the function . In the other case, the impaired renal function lasted until death by heart failure . In the case of abnormal function, discontinuing the ABK therapy improved the hepatic function . In the 4 out of 5 cases that showed excellent or good clinical efficacy, patients recovered within relatively early days of ABK therapy . The average days for recovery was 7.8. Jpn J Antibiot, 1994 Jun, 47(6), 720 - 30 {Clinical efficacy of arbekacin in patients with methicillin-resistant Staphylococcus aureus infections . Research Group of MRSA Forum}; Nasuhara K et al.; We investigated the clinical efficacy of arbekacin (ABK) in patients with methicillin-resistant Staphylococcus aureus (MRSA) infections, and also studied coagulase types, beta-lactamase producing activity and drug sensitivity of MRSA isolated from various clinical specimens . A total of 23 patients with MRSA infections (13 cases of pneumonia, 1 case of sepsis, 1 case of pneumonia and sepsis and 8 cases of the others) who were hospitalized from April 1992 to September 1993 were enrolled in this study . They were 14 males and 9 females, and the mean age was 66.9 years (range, 18-91 years) . All patients had underlying diseases (mainly malignant tumors and cerebrovascular diseases) . ABK was given intravenously at doses ranging from 75 to 100 mg twice daily . The clinical efficacy rate was 90%; 8 cases showed excellent responses, 10 cases good, 1 case fair, 1 case poor and 3 cases were unevaluable . The eradication rate of MRSA was 81.8%; 16 cases were judged as eradicated, 3 cases decreased, 2 cases replaced, 1 case unchanged and in 1 case the bacteriological response was unknown . Side effects were not observed, but S-GPT was elevated in 1 case . Coagulase types of MRSA (123 strains) isolated at the institutes involved in the study were type II (56 strains), type IV (12 strains), type VII (13 strains) and other types (2 strains), but coagulase types of 40 strains could not be determined . Eighty-four strains (68.3%) produced beta-lactamases . MICs of ABK were 0.5 microgram/ml against 43 strains and 1 microgram/ml against 37 strains, and all of the MICs were under 4 micrograms/ml . In summary, ABK showed high antimicrobial activity against MRSA and clinical usefulness in the infections investigated. Jpn J Antibiot, 1994 Jun, 47(6), 701 - 9 {Basic studies on combination effects of arbekacin and beta-lactam antibiotics on methicillin-resistant Staphylococcus aureus}; Watanabe T et al.; Combination effects were studied with arbekacin (ABK) and beta-lactam antibiotics including imipenem/cilastatin (IPM/CS), flomoxef (FMOX), and cefotiam (CTM) for bactericidal activities, post-antibiotic effects (PAE's) and bactericidal activities of beta-lactam antibiotics after removal of ABK using methicillin-resistant Staphylococcus aureus (MRSA) strain 1936 . The following results were obtained . 1 . When ABK was administered in combination with IPM/CS, FMOX or CTM against MRSA strain 1936, low FIC index was not obtained . 2 . Higher bactericidal activity was observed when ABK was given before beta-lactam than when beta-lactam was given before ABK . 3 . Combination of ABK and each of beta-lactam antibiotic led a longer PAE than ABK alone . 4 . When each beta-lactam antibiotic was administered after a treatment and removal of ABK, greater bactericidal activity and growth inhibition were observed than when administered each beta-lactam alone . These findings basically demonstrated that ABK was effectively bactericidal against MRSA when administered in combination with beta-lactam antibiotic such as IPM/CS, FMOX or CTM, even when the FIC index did not indicate a favorable effect. Jpn J Antibiot, 1994 Jun, 47(6), 693 - 700 {Antibiotic efficacies of combined uses of arbekacin and various antibiotics against methicillin-resistant Staphylococcus aureus}; Deguchi K et al.; Efficacies of combined uses of arbekacin(ABK) with other antibiotics against between 57 and 76 clinically isolated methicillin-resistant Staphylococcus aureus(MRSA) were examined . Other antibiotics tested were fosfomycin(FOM), 5 cephems(CEPs), 2 penicillins(PCs) and imipenem . Percentages of strains that grew at various combination of serially diluted drugs in a microdilution method were scored, and for each pair of drugs > or = MIC50 or > or = MIC90 was determined . FIC-indices were calculated at > or = MIC50 . Bacterial growth was inhibited by low concentrations of ABK dose-dependently, while most of the strains tested were resistant to 9 other drugs . FIC-indices with ABK were lowest when used together with ampicillin (ABPC) (0.516) followed by combinations with cefotiam (CTM) and cefuzonam (CZON) (each, 0.53), and FOM (0.625) . When combined with 0.5 microgram/ml of ABK, 0.5 microgram/ml of ABPC inhibited more than 50% of MRSA strains . CTM or CZON in combination with the same amount of ABK to 2 micrograms/ml inhibited more than 50% of MRSA . Thus ABPC was found to be the most efficient in combined use with ABK to inhibit clinical MRSA strains. Jpn J Antibiot, 1994 Jun, 47(6), 640 - 6 Bactericidal activity of arbekacin against methicillin-resistant Staphylococcus aureus . Comparison with that of vancomycin; Aoki Y; Bactericidal activity of arbekacin (ABK) against methicillin-resistant Staphylococcus aureus (MRSA) was compared with that of vancomycin (VCM) . MIC80 of VCM against 1,056 clinical isolates was 2 micrograms/ml and not a single strain of those in which MIC above 4 micrograms/ml was detected . Whereas MIC80 of ABK against the isolates was 1 microgram/ml, but a few strains of which showed MIC of 8 micrograms/ml or 16 micrograms/ml . A killing-curve study indicated that bactericidal activity of ABK was critically dependent on its concentrations . ABK, at higher concentrations, showed excellent killing effects against all the tested isolates, and the effects were superior to those of VCM because of following reasons; great reduction CFU was attained within short time incubation, and the effects were not remarkably influenced by different inoculum sizes of MRSA . At lower concentrations of ABK, between a half and four times MIC, the re-increase of CFU of MRSA with appearance of characteristic small colonies was observed . Considering the concentration of ABK in usual dose, the significance of the re-growth should be carefully assessed . It showed be recommended that the peak concentration of ABK be elevated to higher level if adverse effects do not appear. Jpn J Antibiot, 1994 Jun, 47(6), 634 - 9 {Emergence of arbekacin resistant strains among methicillin-resistant Staphylococcus aureus}; Suzuki T et al.; We studied the efficacy of arbekacin (ABK) against MRSA that were resistant to gentamicin (GM) . ABK was fairly active against most of the GM-resistant MRSA strains . Against highly GM-resistant strains, however, ABK was slightly less active than it was against the low level GM-resistant strains . ABK-resistant mutants were isolated from GM-resistant MRSA at a frequency of 10(-4) to 10(-5) when the selection was made with MIC of ABK . But the frequency decreased with higher concentration of ABK or with a use of 1/2 MIC of beta-lactam antibiotics, such as cefazolin, cefotiam, cefamandole or flomoxef, together with ABK . We isolated a ABK-resistant mutant from a laboratory strain MS353/pMS91 which possess a GM-resistant gene on a plasmid . The gene encodes a modifying enzyme, AAC-6'/APH-2" . We compared the enzyme activities between the mutant MS353/pMS91M and MS353/pMS91 . The mutant strain showed enzyme activity six times as much as the primary strain . This result suggests that the increase of AGs-modifying enzymes was responsible for the ABK-resistance of MRSA. Jpn J Antibiot, 1994 Jun, 47(6), 627 - 33 {Genomic DNA fingerprinting of arbekacin-resistant MRSA by pulsed-field gel electrophoresis}; Nonoyama M et al.; We surveyed 387 clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA) obtained from 26 hospitals isolated in 1993 to determine whether they became resistant to arbekacin (ABK) . Twenty-five ABK-resistant MRSA (6.5%) were isolated from 9 hospitals . Analysis of genomic DNA fingerprinting by pulsed-field gel electrophoresis was used to confirm the classification by resistance patterns, phage typing and other biological characters . After digestion with endonuclease SmaI, two or three types of restriction patterns were found in ABK-resistant MRSA isolated from each hospital . We concluded that ABK-resistant MRSA may spread through nosocomial MRSA infections. Jpn J Antibiot, 1994 Jun, 47(6), 618 - 26 {A survey of Staphylococcus aureus for typing and drug-resistance in various areas of Japan during 1992 and 1993}; Hashimoto H et al.; Data on Staphylococcus aureus strains isolated at various areas in Japan during 1992 and 1993 were collected and analysed . Among 7,033 strains examined, 60.3% were methicillin-resistant S . aureus (MRSA), 86% of which were isolated from inpatients . MRSA were isolated most often (38%) from departments of internal medicine, followed by departments of surgery (13%) . The most frequent source of MRSA was sputa (38%), followed by pus (18%) . As for coagulase types, 69% of MRSA were Type II and next most frequent was Type VII (24%) . Among MSSA strains, Type VII (35%) and Type III (32%) were the most frequent . Frequencies of beta-lactamase-producing strains were 68% in MRSA and 59% in MSSA . More than 80% of MRSA strains were resistant to beta-lactams . The frequencies of resistance to fosfomycin, ofloxacin, minocycline and gentamicin were 88%, 72%, 19% and 66%, respectively . Less than 3% of MRSA strains were resistant to arbekacin or vancomycin. Jpn J Antibiot, 1994 Jun, 47(6), 561 - 74 {Development of arbekacin and synthesis of new derivatives stable to enzymatic modifications by methicillin-resistant Staphylococcus aureus}; Kondo S; Our studies on the resistance mechanisms and chemical modifications of aminoglycoside antibiotics led to the synthesis of arbekacin (ABK), which was refractory to most aminoglycoside-modifying enzymes in resistant bacteria . In 1990, ABK was launched into clinical uses in Japan as a chemotherapeutic agent for the treatment of infections caused by methicillin-resistant Staphylococcus aureus (MRSA) . By 1993 only a few MRSA strains moderately resistant to ABK (MIC, 6.25-12.5 micrograms/ml) had clinically been isolated . ABK was modified by the reaction with an excess of an enzyme preparation extracted from an ABK-resistant strain (12.5 micrograms/ml) and three inactivated products were produced, consisting mainly of ABK 2''-phosphate along with small amounts of 6'-N-acetyl-ABK and the doubly modified ABK . Based on these results, replacement of the 2''-hydroxyl by amino group in dibekacin (DKB) or in ABK was designed to obtain potent active derivatives against MRSA . Conversion of the 2''-hydroxyl group by DMSO-DCC oxidation followed by reductive amination with NH4OAc-NaBH3CN gave 2''-amino-2''-deoxy-DKB (D1) and -ABK (A1) . Their 5-deoxy (D2 and A2), 5-epifluoro (D3 and A3) and 5-epiamino (D4 and A4) derivatives were also synthesized . All 2''-amino-2''-deoxy-ABK derivatives (A1, A2, A3 and A4) showed excellent activities against MRSA and Gram-negative bacteria, as expected . Among them, A4 having low acute toxicity and nephrotoxicity was selected as a new candidate for anti-MRSA agent. APMIS, 1994 Jun, 102(6), 407 - 12 Evaluation of methods for the detection of nasal carriage of Staphylococcus aureus; Eriksen NH et al.; In the present study we investigate the optimal methodology for determination of the nasal carriage rate of Staphylococcus aureus . Tests were performed on 91 healthy laboratory staff . The reproducibility of different sampling, transportation, storage and culture methods was examined . We compared sterile dry cotton wool swabs with sterile dry cotton wool swabs impregnated with charcoal and 5% blood agar plates with mannitol salt agar plates after different incubation periods . Finally, we investigated the detection rate for S . aureus following direct plating compared to storage in Stuart's transport medium for 7 days . There were no differences in isolation rates from the right or left nostril using either cotton or charcoal swabs . Charcoal swabs gave an increased isolation rate as compared to cotton swabs, and incubation in broth enrichment medium containing 6.5% NaCl also increased the isolation rate . Storage in Stuart's transport medium for 7 days gave an increase in isolation rate as compared to direct plating on blood agar . With mannitol salt agar plates the increase in isolation rate when incubation was performed for from 2 to 4, 2 to 7, and 4 to 7 days was 5.9%, 16.7%, and 11.5%, respectively . For the detection of S . aureus nasal carriers we find the use of charcoal swabs and Stuart's transport medium combined with cultivation on mannitol salt agar for 7 days to be the optimal method. W V Med J, 1994 Jun, 90(6), 238 - 41 Staphylococcus aureus: a continuing problem; Neely JL; Caution is required in managing any immunocompromised host, not only because these patients will be carriers, but because they are also very susceptible to infections with S . aureus . These hosts are not candidates for short-course antibiotic therapy, and catheters should be removed when S . aureus bacteremia is diagnosed . The S . aureus cell wall is a major determinant of the host response and the pathogenicity of this organism . The clinician should recognize the three most important toxins produced by S . aureus: exfoliatin, TSST-1, and enterotoxin-B . Toxic shock syndrome can occur in any host, not just menstruating females, and the clinician should be very thoughtful when dealing with any Staphylococcus aureus infections arising from the use of a catheter. Artif Organs, 1994 Jun, 18(6), 448 - 53 Reduced susceptibility of polytetrafluoroethylene vascular prostheses to colonization by Staphylococcus aureus following in situ endothelialization; Zdanowski Z et al.; The impact of endothelialization of polytetrafluoroethylene (PTFE) grafts on susceptibility to experimental colonization by Staphylococcus aureus was studied in a rat model . One hundred and eight grafts (2 mm inner diameter, 5 mm length) were implanted into the infrarenal aorta (54 rats) or the infrarenal caval vein (54 rats) . The progress of endothelialization following graft implantation was evaluated by SEM at 1, 3, 7, and 14 days on 6 grafts from each group . We found that the endothelialization was more rapid in caval vein than in aorta: The caval vein grafts were completely endothelialized 2 weeks following implantation compared with endothelialization of approximately 0.5 mm of aorta grafts, measured from each anastomosis . During this time, the resistance to standardized intravenous challenge with 10(8) colony-forming units of S . aureus increased gradually in both groups, and all caval vein grafts tested at 2 weeks were sterile . However, all aorta grafts challenged at 2 weeks were colonized, although to a somewhat lower degree than at earlier challenge . Resistance of PTFE grafts to colonization with S . aureus thus correlated to the degree of endothelialization. Int J Pediatr Otorhinolaryngol, 1994 Jun, 29(3), 249 - 55 Septic cavernous sinus thrombosis following infection of ethmoidal and maxillary sinuses: a case report; Assefa D et al.; A 10-year-old boy with a unilateral septic cavernous sinus thrombosis complicating infection of the ethmoidal and maxillary sinuses is described . The causative agents identified were Eikenella corrodens and Staphylococcus aureus . The disease was cured without any sequelae. Kansenshogaku Zasshi, 1994 Jun, 68(6), 740 - 3 {Isolation of methicillin resistant Staphylococcus aureus from nasopharyngeal swabs on admission to a ward for pediatric patients}; Sakata H et al.; Between 1992 and 1993, 2313 nasopharyngeal swabs were taken and cultured from 1665 patients on admission to a pediatric ward at Asahikawa Kosei Hospital . Twenty nine strains of Methicillin resistant Staphylococcus aureus (MRSA) were isolated from 28 patients . The ages of these patients ranged from 2 months to 9 years and 14 patients were younger than 1 year . Their underlying diseases were bronchial asthma in 9 patients, prematurely born infants with chronic lung disease in 4 . Fourteen patients had a history of admission during the past 6 months . No patient had infectious diseases due to MRSA. Br J Surg, 1994 Jun, 81(6), 872 - 4 Conservative initial treatment for liver abscesses in children; Moore SW et al.; A total of 124 children aged less than 14 years with a liver abscess were seen in a 16-year period (1974-1990) and treated by non-operative initial management . Of the abscesses 98 occurred in the right liver and 26 in the left . The abscesses were solitary in 93 patients . Overall, 77 of the solitary and 21 of the multiple abscesses were confined to the right liver . In 78 of the right-sided and 20 of the left-sided abscesses the infection was primarily pyogenic in nature with Staphylococcus aureus being the usual organism cultured . The remainder were of amoebic origin . Clinical features were similar in patients with amoebic and pyogenic abscesses . Clinical and ultrasonographic follow-up demonstrated successful non-operative management and healing in 37 per cent of all patients submitted to an initial protocol of medical supportive care and antibiotic therapy . Of the multiple abscesses 60 per cent responded to non-operative management . Fourteen of the 16 solitary left-sided liver abscesses required drainage and three left-sided abscesses ruptured before drainage . Patients with a solitary left-sided abscess warrant early operative intervention. FEMS Microbiol Lett, 1994 Jun 1, 119(1-2), 59 - 63 Biochemical and genetic heterogeneity of staphylococcal protein A; Sakurada J et al.; We investigated the biochemical and genetic heterogeneity of protein A from Staphylococcus aureus . SpA genes (spas) of various strains were heterogeneous when detected as DraI and EcoRV fragments of chromosomal DNA . Polymerase chain reaction using primers to detect DNA encoding the IgG-binding domains in spa revealed that they numbered between 2 and 5 . Protein A from several S . aureus strains showed two types of reactivities to immunoglobulins in normal canine serum according to the number of active domains. FEMS Microbiol Lett, 1994 Jun 1, 119(1-2), 167 - 73 Typing of Staphylococcus aureus by amplification of the 16S-23S rRNA intergenic spacer sequences; Dolzani L et al.; The possibility of using polymorphisms in the spacer regions between 16S and 23S rRNA genes in order to type Staphylococcus aureus has been evaluated . To this purpose, DNA extracted from 74 independent isolates was amplified making use of a pair of primers complementary to conserved regions in the 16S and 23S genes . We have demonstrated that the method provides a good discrimination between unrelated isolates, giving better results when methicillin-sensitive strains are considered . Moreover, the amplification profiles were reproducible and all strains were typable . Given these results, and the technical simplicity of the process, we propose PCR-ribotyping to be taken into consideration as a method for typing S . aureus. Am J Physiol, 1994 Jun, 266(6 Pt 1), C1673 - 83 Metabolic characteristics of alpha-toxin-permeabilized smooth muscle; Trinkle-Mulcahy L et al.; Rabbit portal veins were permeabilized using Staphylococcus aureus alpha-toxin, and adenosinetriphosphatase (ATPase) was measured as the formation of {3H}ADP, {3H}AMP, and {3H}adenosine from {3H}ATP in the solution bathing the muscle . The resting ATPase (1.96 +/- 0.15 mM/min, n = 13) is approximately 5-10 times higher than that measured in Triton X-100-permeabilized muscles (0.28 +/- 0.01 mM/min, n = 4), with nucleotide accumulating as ADP, AMP, and adenosine . The ATPase activity is also seen when the intact muscle is incubated in a Krebs solution containing 1 mM MgATP (2.76 +/- 0.10 mM/min, n = 73) . This suggests that it is due primarily to an ecto-ATPase . The ectoenzyme is capable of hydrolyzing both ATP and ADP, and in both cases there is a higher rate at 3 than at 1 mM nucleotide . The high resting ATPase compromises the control of nucleotide concentrations within the permeabilized tissue even in the presence of an ATP-regenerating system consisting of phosphocreatine (PCr, 35mM) and creatine kinase (1 mg/ml) . Treatment of the intact muscle with the ectonucleotidase inhibitor 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS) followed by alpha-toxin permeabilization and inclusion of sodium azide in subsequent solutions reduces the ecto-ATPase by approximately 70% . Addition of PCr and creatine kinase then results in the maintenance of high {ATP} and low {ADP} in the muscle, and importantly, there are no significant changes in {ATP}, {ADP}, {adenosine/AMP}, or the ADP-to-ATP ratio upon activation of the muscle in pCa 4.5 . In general, the force output in high Ca2+ increased as the metabolic profile of the muscle improved . When ATPase was measured as the appearance of {32P}Pi from {32P}PCr and {gamma-32P}ATP, the alpha-toxin-permeabilized muscle subjected to the above treatment showed only approximately 30% higher total ATPase under activated conditions compared with the freeze-glycerinated Triton-treated portal vein . The suprabasal ATPase is similar in both preparations . We conclude that the reduction of the basal ATPase by the DIDS-azide treatment permits both rigorous control of nucleotide contents and accurate measurement of ATPase activity in alpha-toxin-permeabilized smooth muscle. Epidemiol Infect, 1994 Jun, 112(3), 501 - 11 Clonal study of enterotoxin-B producing strains of Staphylococcus aureus; Renaud F et al.; Sixty-nine Staphylococcus aureus strains, 39 of which produced staphylococcal enterotoxin B (SEB+) and 14 of which were associated with toxic shock (TS+), were studied using the following markers: serotyping, phage typing, antibiotyping, ribotyping, zymotyping and pulsed-field electrophoresis typing . Analysis of the results showed that the enterotoxin B producing strains were derived from at least three clones: the first two consisted of methicillin-susceptible strains, while the third included the methicillin-resistant (MRSA) strains . TS+ strains of nongenital origin appeared to be distributed between the three clones, with no specific characters. Epidemiol Infect, 1994 Jun, 112(3), 489 - 500 Epidemiology of Staphylococcus aureus in patients with cystic fibrosis; Branger C et al.; Seven hundred and thirty-four isolates of Staphylococcus aureus, recovered from the sputum of 238 cystic fibrosis patients in six French hospitals, were characterized by esterase electrophoretic typing, capsular polysaccharide serotyping and phage typing and tested against 14 antibiotics for sensitivity . Thirty-four esterase electrophoretic types were found with a genotypic diversity coefficient of 0.91 . Five hundred and forty-eight (78.7%) isolates produced capsular polysaccharide and 350 (50.3%) were type 8 . Four hundred and sixty isolates (66.6%) were phage typable and 202 (28.2%) were lysed by group III bacteriophages . No esterase electrophoretic type, capsular type or phage type was specific to cystic fibrosis . Isolates belonged to a wide range of types, similar to strains acquired outside hospitals . Eighty-five patients had three or more consecutive isolates over at least 6 months . The ability of S . aureus to persist for long periods of time has been demonstrated in 73% of them . Methicillin-resistance was encountered among 73 strains (9.8%) which were also multiresistant . Two hundred and eighty-nine (39.9%) strains were sensitive to all antibiotics tested except to penicillin . Pristinamycin and co-trimoxazole were the most effective antibiotics . These results could contribute to the elaboration of a rational approach to the prophylaxis and therapy of respiratory staphylococcal infections in cystic fibrosis patients. Clin Exp Immunol, 1994 Jun, 96(3), 508 - 12 Effects of cyclosporine and rapamycin on immunoglobulin production by preactivated human B cells; Kim HS et al.; In order to assess the direct effects of cyclosporine A (CsA) and rapamycin on B cells, we utilized a two-segment culture system of highly purified B lymphocytes consisting of induction (activation) in the presence of the formalinized Staphylococcus aureus bacteria and IL-2, and differentiation, respectively, in the presence of various combinations of cytokines . Results show that rapamycin strongly inhibited production of both IgM and IgG measured at the end of the secondary culture supported by IL-2/IL-6, whereas CsA up-regulated the immunoglobulin production . The stimulatory effect of CsA was also observed when preactivated B cells were recultured in absence of any cytokines . These results show that rapamycin and CsA have clearly distinct effects on human B lymphocyte responses in vitro . Rapamycin is a more potent in vitro immunosuppressant of B lymphocytes than CsA . It is effective at significantly lower concentrations, and it does not stimulate either the proliferation or antibody production by preactivated B cells. Arthritis Rheum, 1994 Jun, 37(6), 942 - 50 Comparative inhibitory effects of bucillamine and D-penicillamine on the function of human B cells and T cells; Hirohata S et al.; OBJECTIVE . Clinical trials have suggested that the efficacy of bucillamine (BUC) in rheumatoid arthritis (RA) may be superior to that of D-penicillamine (DP), although the basis of the differences remains unclear . Previous studies have revealed that BUC has unique immunomodulatory effects that depend upon its capacity to form an intramolecular disulfide (BUC-ID) . We therefore examined the effects of BUC-ID on the in vitro function of human B cells and T cells compared with those of DP, at their pharmacologically attainable concentrations . METHODS . IgM production was induced in highly purified B cells from healthy donors by stimulation with Staphylococcus aureus Cowan 1 (SAC) plus interleukin-2 (IL-2) or with immobilized anti-CD3-activated CD4+ T cells . Interferon-gamma (IFN gamma) production was induced in CD4+ T cells by stimulation with immobilized anti-CD3 . RESULTS . BUC-ID suppressed IgM production induced by SAC+IL-2 as well as that induced by immobilized anti-CD3-activated CD4+ T cells, whereas DP suppressed the latter more markedly than the former . DP (3 micrograms/ml) significantly suppressed IFN gamma production by immobilized anti-CD3-stimulated CD4+ T cells, but not IgM production induced by SAC + IL-2 stimulation . By contrast, BUC-ID (0.3 microgram/ml) significantly suppressed IgM production induced by SAC + IL-2, but not T cell IFN gamma production . Of note, BUC-ID did not suppress IL-6 production by SAC-activated B cells . CONCLUSION . These results indicate that the target cells of BUC and DP in vivo might be different, with the former inhibiting the function of B cells and the latter that of T cells . The data suggest the possibility that BUC may have a different effect in RA patients compared with the effect of DP, and may be effective in patients who do not respond to DP. Singapore Med J, 1994 Jun, 35(3), 277 - 82 An outbreak of methicillin-resistant Staphylococcus aureus in a neonatal intensive care unit in Singapore: a 20-month study of clinical characteristics and control; Tan KW et al.; Methicillin resistant Staphylococcus aureus (MRSA) is a major infection control problem in many countries . There have been many reports of outbreaks in neonatal nurseries including, in our part of the world, Australia, Hong Kong and Malaysia . A recent outbreak of MRSA in the neonatal intensive care unit in the Kandang Kerbau Hospital, Singapore, presented us with the opportunity to study the clinical characteristics of the outbreak and the effects of infection control measures . Neonates admitted to the neonatal intensive care unit were studied over a 20-month period . They were all screened for nasal colonisation on admission and weekly thereafter . Infections were documented . Over this period there were altogether 2,576 admissions of which 85 infants had nasal colonisation with MRSA (3.3%) and 28 developed infections (1%) . Although the majority of infants colonised by MRSA suffered no ill effects, 3 had septicaemia and 2 had septicaemia with osteomyelitis . There were no deaths . Standard infection control measures with barrier nursing and the use of mupirocin nasal ointment were ineffective, and control was achieved only after strict cohorting together with the use of mupirocin was instituted . This was done without additional costs to the department and without additional nurses or doctors. Mol Biochem Parasitol, 1994 Jun, 65(2), 259 - 70 Expression of a hydrophilic surface protein in infective stages of Leishmania major; Flinn HM et al.; A family of differentially expressed genes from Leishmania major contains one sequence (Gene B) that encodes a novel, hydrophilic protein found on the surface of infective parasite stages . The 177-residue, acidic Gene B protein is characterised by an amino acid repetitive element, comprising 45% of the total molecule, that is related to the cell-wall binding domain of protein A from Staphylococcus aureus . No identifiable signal peptide, membrane-spanning domain or consensus for glycosylphosphatidylinositol anchor attachment to the cell surface is found elsewhere in the deduced protein sequence . In vitro, the Gene B protein fractionates with the parasite cell surface glycoconjugates, lipophosphoglycan and the glycoinositolphospholipids . This protein is the first characterised surface peptide marker for infective stages of the Leishmania life cycle. J Electron Microsc (Tokyo), 1994 Jun, 43(3), 164 - 7 Distribution of WGA-binding sites on the surface of clinical isolates of Staphylococcus aureus; Yamada S et al.; Distribution of wheat germ agglutinin (WGA)-binding sites on the surface of clinical isolates of Staphylococcus aureus was examined by WGA-gold . Labelings on three methicillin-sensitive S . aureus (MSSA) isolates were heavier than those on three methicillin-resistant (MRSA) isolates and the 209P (MSSA) strain . These results were confirmed by an alkaline phosphatase-WGA assay . The WGA-binding on the MRSA was consistently poor, whereas wide ranging diversity was observed in WGA-binding among the MSSA . These results strongly suggest diversities in not only distribution, but also the quantity of WGA-binding carbohydrates exposed on the surface of the organisms. Am J Infect Control, 1994 Jun, 22(3), 152 - 62 Feasibility of a combined carrier test for disinfectants: studies with a mixture of five types of microorganisms; Best M et al.; BACKGROUND: There is mounting concern regarding the efficacy of many germicides on the market because officially recognized germicidal tests for various classes of microorganisms vary widely and often lack reproducibility and proper quantitation . We report here a carrier method for simultaneously and quantitatively assessing the efficacy of liquid chemical germicides against a mixture of microorganisms of varying degrees of resistance . METHODS: In the test, each small glass cup (10 mm wide x 14 mm long) was contaminated with 10 microliters of a standardized mixture of Staphylococcus aureus, Mycobacterium bovis bacille Calmette-Guerin, Trichophyton mentagrophytes spores, Sabin poliovirus type 1, and Bacillus stearothermophilus spores in 5% fetal bovine serum . The inoculum was dried for 60 minutes under ambient conditions and covered with 60 microliters of the disinfectant under test or a balanced salt solution control for the desired contact time . The carrier was then placed in 2940 microliters of an eluent and the eluates assayed separately for the five microorganisms . Tap water was used to dilute the test product as needed . RESULTS: Of the 11 products tested, 2% alkaline glutaraldehyde, 0.6% sodium hypochlorite (about 5000 ppm free chlorine), and a 0.4% quarternary ammonium compound containing 23% hydrochloric acid were effective against all five challenge organisms . A hard-surface spray containing 0.1% o-phenylphenol with 79% ethanol was effective against all but bacterial spores; 70% (volume/volume) ethanol alone and povidone-iodine (1% available iodine) were effective against S . aureus, the mycobacterium, and the fungus; a 3% solution of peroxygen compounds was effective only against S . aureus and the poliovirus; 1.5% chlorhexidine gluconate, 0.06% quaternary ammonia compound, and 0.03% o-phenylphenol + 0.03% p-tertiary amylphenol could inactivate nothing but S . aureus; and 3% hydrogen peroxide was ineffective in all tests . CONCLUSIONS: This method shows promise for use with various classes of microorganisms, individually or as mixtures . Its application should enable the classification of germicides according to spectrum of activity. Acta Trop, 1994 Jun, 57(1), 11 - 20 The sensitivity of axenic 200:NIH Entamoeba histolytica to bacitracin and its zinc salt; Andrews BJ; This study extends the previous knowledge of the action of bacitracin zinc against xenic cultures of Entamoeba histolytica . Results presented here, using axenic E . histolytica, confirm the enhanced activity of bacitracin zinc as compared to native bacitracin and demonstrate that this enhancement is due to a direct effect upon the trophozoite . In the presence of zinc, amoebastatic concentrations of bacitracin became amoebacidal . The expression of the amoebacidal characteristics of 5 g/l bacitracin zinc following 24-35 h incubation coincided with the return of cell division by amoeba exposed to the same concentration of native bacitracin . The activity of both bacitracin and bacitracin zinc on trophozoites was unaffected by autoclaving, indicating a mode of action different from that exerted on Staphylococcus aureus . In contrast to previous clinical observations, no synergism could be detected between bacitracin and neomycin. Khirurgiia (Mosk), 1994 Jun, (6), 32 - 5 {Microbiology of suppurative wound in laser radiation}; Cherkasskaia RS et al.; Comparative quantitative and qualitative appraisal of purulent lesions of the soft tissues in surgical patients treated with the infrared lasers "Uzor" (pulse) and "Skalyar (continuous) and by the traditional methods (antibiotics, antiseptics, ointments) was conducted under clinical conditions . Abscesses, mastitis, and phlegmons were studied before and after the operation and on the 3rd, 6th, and 12th postoperative day . It was found that Staphylococcus aureus in a monoculture was the etiological cause of purulent infection in 75.2% of cases . The bactericidal effect of pulse and continuous infrared irradiation led to a decrease in the number of infected patients and more rapid cleansing of the wound . In treatment with Skalyar the focus of affection was cleaned completely by the 6th day . Qualitative improvement of the cultural, biochemical, and lysogenic properties and antibiotic sensitivity of the staphylococcus isolated from patients in dynamics during irradiation did not occur. J Hosp Infect, 1994 Jun, 27(2), 139 - 47 Surveillance of postoperative infections in thoracic surgery; Kluytmans JA et al.; Postoperative infections (PIs) are serious complications of thoracic surgery . To gain insight into the nature and the scope of the problem, an 18-month prospective surveillance was conducted at the department of thoracic surgery of the University Hospital Rotterdam, Dijkzigt . PI were classified according to CDC criteria . One hundred and ninety-four out of 983 patients (19.7%) developed one or more PIs and in these 194 patients, 268 PIs were diagnosed . The incidence of PI was 2.0 per 100 days of postoperative stay . The mean postoperative length of stay (LOS) of the 194 patients with PI was 14.1 days longer than those without PI . Deep surgical wound infections (DSWIs) were associated with the longest prolongation of the median postoperative LOS in the hospital (30 days longer) . Although lower than DSWIs, incisional surgical wound infections also had a significant prolongation of stay (median 10 days longer) . Staphylococcus aureus was the most important pathogen associated with surgical wound infections (SWIs) . Phage typing of 29 strains causing SWI showed only two identical pairs, so only a minority of infections could be explained by cross-infection . Older age, and more complicated procedures (e.g . cardiac valve operations) were independent, statistically significant, risk factors for the development of PI . Since there is a progressive trend towards operating on older patients and performing more complicated procedures, the incidence of PI is expected to increase . Therefore it will become increasingly important to develop new strategies to prevent these serious complications. J Hosp Infect, 1994 Jun, 27(2), 127 - 34 Nasal carriage of Staphylococcus aureus in Australian (pre-clinical and clinical) medical students; Stubbs E et al.; The nasal carriage of Staphylococcus aureus in 808 Australian medical students was studied . Five groups of students experienced varying degrees of clinical exposure in a hospital environment ranging from 0 to 42 months . The overall percentage of carriers among the five groups did not vary . However, with increasing clinical exposure there was a decrease in the percentage of isolates sensitive to all antibiotics tested, and an increase in the carriage of S . aureus resistant to three or more antibiotics . No carriers of methicillin-resistant S . aureus (MRSA) were detected . The comparative rates of S . aureus carriage between female and male students varied . The relevance of medical students as nasal carriers of S . aureus in the hospital environment today is discussed. J Antimicrob Chemother, 1994 Jun, 33(6), 1191 - 200 Comparative efficacies of imipenem, oxacillin and vancomycin for therapy of chronic foreign body infection due to methicillin-susceptible and -resistant Staphylococcus aureus; Schaad HJ et al.; The efficacies of imipenem when directed against methicillin-susceptible (MSSA) and methicillin-resistant (MRSA) strains of Staphylococcus aureus were compared with those of oxacillin and vancomycin in a subcutaneous rat model, using chronically infected tissue cages . At three weeks after inoculation, stable chronic infections were established with average bacterial counts exceeding 10(6) cfu/mL tissue cage fluid for both strains . Intraperitoneal administration (twice a day for 7 days) of imipenem (80 mg/kg) or oxacillin (200 mg/kg) produced peak levels of 23 or 45 mg/L and through levels of < 0.1 and 5.7 mg/L, respectively . The therapeutic regimens of either imipenem (P < 0.001) or oxacillin (P < 0.02) administered for 7 days led to significant reductions in bacterial counts in the tissue cage fluids of animals chronically infected with MSSA . In contrast, imipenem was not effective against chronic MRSA tissue cage infections, despite the relatively low MIC of the infecting strain and the use of high dose (120 mg/kg) therapy . In-vitro susceptibility testings of MRSA performed before and after imipenem therapy demonstrated the emergence of a highly resistant subpopulation. J Antimicrob Chemother, 1994 Jun, 33(6), 1165 - 71 The accumulation of novobiocin by Escherichia coli and Staphylococcus aureus; Barrett-Bee K et al.; The accumulation of novobiocin by Escherichia coli was shown to be a nonsaturable process . Inhibitors or uncouplers of respiration had no effect on accumulation in cells grown in Isosensitest broth, whereas, bacteria grown in nutrient broth demonstrated an energy-dependent efflux mechanism . In contrast, accumulation of novobiocin by Staphylococcus aureus was found to have a Km of 74 microM and uncouplers and inhibitors of respiration reduced the accumulation suggesting that in S . aureus novobiocin is transported actively. J Antimicrob Chemother, 1994 Jun, 33(6), 1155 - 63 Effect of combination of oxacillin and non-beta-lactam antibiotics on methicillin-resistant Staphylococcus aureus; Komatsuzawa H et al.; The in-vitro activity of oxacillin combined with non-beta-lactam antibiotics, bacitracin, vancomycin, enduracidin, tunicamycin, flavomycin, fosfomycin, and cycloserine, was investigated in 23 methicillin-resistant and 15 methicillin-susceptible Staphylococcus aureus strains . In the presence of a non-beta-lactam antibiotic (0.25 MIC), the MICs of oxacillin for methicillin-resistant S . aureus (MRSA) strains and methicillin-susceptible S . aureus (MSSA) strains were lowered . This effect was most marked with MRSA strains, and bacitracin, flavomycin, and tunicamycin increased the susceptibility of MRSA to oxacillin by greater than 200-fold . More than 87% and 77% of MRSA and MSSA strains, respectively, were synergically inhibited by a combination of oxacillin with bacitracin, tunicamycin, flavomycin or cycloserine (fractional inhibitory concentration < or = 0.5) . Polyanetholesulfonic acid prevented the lysis of MRSA cells treated with a combination of oxacillin and 0.25 MIC of bacitracin, but did not prevent cell death . The penicillin binding protein (PBP) profile of MRSA was not affected by incubation with 0.25 MIC of non-beta-lactam antibiotics . These results suggest that the increased antibacterial activity of oxacillin in the presence of non-beta-lactam antibiotic is not the consequence of activation of autolysis or of a decrease in bulk PBP synthesis. Herz, 1994 Jun, 19(3), 149 - 51 {Aortic and mitral valve endocarditis after infection of the pacemaker pocket}; Funck R et al.; The case of a 76-year-old diabetic patient with known aortic valve sclerosis is reported . One week after implantation of a permanent pacemaker system (indication: 2nd degree AV-block type Mobitz) he developed fever . Large endocarditic vegetations were found on the aortic and mitral valve (blood cultures: were positive for Staphylococcus aureus) . Also from the pacemaker bed Staphylococcus aureus was isolated and an antibiotic treatment including vancomycin was started . Nevertheless the patient developed insufficiencies of both the aortic and mitral valves and became hemodynamically unstable . Due to cerebral embolisms and further deterioration of the patient's overall clinical state the already planned operative replacement of the aortic and mitral valve could not be performed . The patient died because of left ventricular failure after pacemaker infection which was complicated by endocarditis. Herz, 1994 Jun, 19(3), 144 - 8 {Fever and lung abscesses in anorexia nervosa after infusion therapy}; Hoffmann J et al.; A 26 year old female patient was admitted to our hospital because of septic temperatures and chills . In the patient's history renal insufficiency has been known for several years due to agenesia of the right and pyelonephritic renal congestion of the left kidney . Long lasting anorexia nervosa had been treated by psychotherapeutical interventions for years and when failing it necessitated repeated intravenous nutrition by central venous lines . The prominent symptom of the intravenously treated young woman was fever up to 39.7 degrees C and pneumonia, which was considered by the first treating clinic to be caused directly by diminished immunoreactivity in malnutrition and preuremia . The chest X-ray confirmed pneumonia and revealed multiple abscesses in both lungs (Figure 1) . After being transferred to our intensive care unit the pathophysiological context became obvious . From inspection (positive jugular pulsation), from auscultation (holosystolic murmur at the left parasternal border) tricuspid incompetence due to infective endocarditis was suspected . This was confirmed immediately by TM and two-dimensional transthoracic echocardiography, which showed a large vegetation on the anterior tricuspid valve leaflet (Figures 2a and 2b) . Tricuspid regurgitation was also ascertained by color flow echocardiography (Figure 2c) . Several blood cultures were positive for staphylococcus aureus . Clinical and laboratory recovery was achieved by antibiotic therapy with vancomycin and cephtazidim for 3 months.(ABSTRACT TRUNCATED AT 250 WORDS) FEMS Immunol Med Microbiol, 1994 Jun, 9(1), 35 - 9 Construction of a gyrA plasmid for genetic characterization of fluoroquinolone-resistant Staphylococcus aureus; Tankovic J et al.; We have constructed a gyrA trans-complementation plasmid, pAT512, by cloning the wild-type gyrA gene of Staphylococcus aureus into the expression vector pAT392 . Introduction by electrotransformation of pAT512 into a high-level fluoroquinolone resistant mutant of S . aureus (ciprofloxacin MIC = 16 micrograms ml-1) having a gyrA mutation which results in a Ser-84 to Leu substitution, reduced the MICs of norfloxacin and ciprofloxacin for the host of four- and eight-fold, respectively. Eur J Epidemiol, 1994 Jun, 10(3), 325 - 30 Molecular epidemiology for local outbreaks of methicillin resistant Staphylococcus aureus (MRSA) . The need for several methods; Sabria-Leal M et al.; Subtyping isolates may be useful for epidemiological studies of methicillin-resistant-Staphylococcus aureus (MRSA) outbreaks . Among subtyping methods, DNA-based techniques have been applied very effectively for this purpose . An outbreak of MRSA infections took place in one hospital in Barcelona early during 1991 . From the beginning of the outbreak to December 92, 70 MRSA isolates from different patients and sources were collected . All strains were evaluated by restriction endonuclease analysis of plasmid DNA (REAP) and macrorestriction endonuclease analysis of genomic DNA using Sma I and pulsed-field-gel-electrophoresis (PFGE) . Plasmid screening and REAP using Hind III demonstrated two plasmid subtypes: subtype A showing a large plasmid, and subtype B showing the same large plasmid plus a smaller one . Subtypes A and B corresponded to the more recent and older isolates, respectively, suggesting the loss of the small plasmid during the epidemic . PFGE using Sma I displayed two closely related profiles (PFGE subtype A and A'; CS = 0.90) . These subtypes were different from those subtypes exhibited from 4 methicillin-susceptible-Staphylococcus aureus (MSSA) isolates from the same hospital and from 2 epidemiologically unrelated MRSA isolates . Almost all isolates showing PFGE subtype A preceded those isolates showing PFGE subtype A' . This fact and the similarity between both subtypes suggested minor chromosomal DNA rearrangement during the outbreak from a unique strain . While PFGE using Sma I is a useful tool in evaluation of clonal dissemination, our data suggest epidemic or local outbreaks may need several methods to best delineate the source and spread of MRSA strains . The reproducibility and discriminatory power of REAP makes it a useful adjunct in this context. J Immunol, 1994 Jun 1, 152(11), 5259 - 67 Prostaglandin E2 inhibits T cell-dependent Ig secretion by neonatal but not adult lymphocytes; Splawski JB et al.; Prostaglandin E2 (PGE2) inhibits Ig production by adult B cells stimulated with Staphylococcus aureus and IL-2, and inhibits the production of IL-2 by anti-CD3 stimulated T cells . By contrast, PGE2 did not inhibit T cell-dependent Ig production by adult B cells . However, T cell-dependent Ig secretion by neonatal B cells was markedly inhibited by PGE2 even in the presence of supplemental IL-2 . Forskolin, a direct activator of adenylate cyclase, and dibutyryl cAMP caused similar effects . PGE2 inhibited T cell-dependent Ig secretion by both neonatal CD5+ and CD5- B cells but did not inhibit Ig secretion by either CD5+ or CD5- adult peripheral blood B cells . PGE2 did not inhibit IL-2-dependent anti-CD3 stimulated neonatal T cell proliferation or T cell-dependent neonatal B cell proliferation . PGE2 inhibited neonatal B cell Ig production when addition was delayed, suggesting that initial B cell activation and proliferation were not blocked, but that PGE2 prevented subsequent differentiation . In addition, PGE2 inhibited neonatal T cell help independent of cytokine production . PGE2-induced cAMP levels in neonatal B and T cells were not different from adult levels . These results indicate that production of Ig by neonatal lymphocytes is uniquely sensitive to the inhibitory effects of agents such as PGE2 that elevate cAMP levels . This sensitivity may contribute to the suppressed in vivo responses of human neonatal B cells. Infect Immun, 1994 Jun, 62(6), 2529 - 35 Leukotriene B4 generation and DNA fragmentation induced by leukocidin from Staphylococcus aureus: protective role of granulocyte-macrophage colony-stimulating factor (GM-CSF) and G-CSF for human neutrophils; Hensler T et al.; We studied the effect of leukocidin from Staphylococcus aureus V8 strains (Luk-PV) on the generation of Leukotriene B4 (LTB4) and its metabolites from human polymorphonuclear neutrophils (PMNs) . Significant amounts of LTB4 were generated by PMNs after leukocidin exposure in a time- and dose-dependent manner, as shown by reversed-phase high-performance liquid chromatography analysis . In this regard, the S and F components of leukocidin acted synergistically . The calcium ionophore A23187 induced LTB4 generation, and the metabolism of exogenously added LTB4 into biologically less active omega-oxidated compounds was significantly decreased after leukocidin exposure . Priming of PMNs with granulocyte-macrophage colony-stimulating factor (GM-CSF) or G-CSF prior to leukocidin exposure substantially increased toxin- and calcium ionophore A23187-induced LTB4 formation . The inhibitory effects of leukocidin on mediator release were accompanied by membrane damage and DNA fragmentation, which were both restored after pretreatment with GM-CSF . The data suggest that the presence of costimulatory priming factors such as GM-CSF or G-CSF in the microenvironment of an inflammatory focus determines the pathophysiological effects induced by S . aureus leukocidin. Nature, 1994 May 26, 369(6478), 324 - 7 Binary and ternary complexes between T-cell receptor, class II MHC and superantigen in vitro; Seth A et al.; Superantigens are proteins that in association with class II major histocompatibility complex (MHC)-bearing cells can stimulate virtually all T cells that express particular classes of the variable beta-domains of the T-cell receptor (TCR) . This mechanism of T-cell activation circumvents the usual requirement for peptide-specific MHC recognition . Staphylococcus aureus enterotoxin B (SEB) is a bacterial superantigen that causes food poisoning and shock . We have characterized the tertiary complex of SEB, a soluble T-cell receptor, and a soluble class II MHC molecule DR1, and the three binary complexes TCR-SEB, SEB-DR1, and the peptide-specific complex DR1-TCR . We report here that in each case the specificity of the interaction among the soluble molecules is the same as observed in biological assays . Native gel electrophoresis and plasmon resonance affinity measurements indicate that SEB-TCR complex can form in the absence of class II MHC and that SEB-TCR interaction increases the binding of DR1 . The observation that a superantigen can form complexes with TCR in both the absence and presence of class II MHC may provide a mechanism for its ability to induce anergy in some circumstances and activation in others (reviewed in ref . 8). FEMS Microbiol Lett, 1994 May 15, 118(3), 199 - 205 Surface-associated proteins of Staphylococcus aureus: their possible roles in virulence; Foster TJ et al.; A class of proteins that are associated with the cell surface of Gram-positive bacteria has been recognised . Common structural features which are implicated in the proper secretion and attachment of these proteins to the cell surface occur in the C-termini . N-terminal domains interact with the host by binding to soluble host proteins, to matrix proteins or to host cells . They probably have important roles in pathogenicity by allowing bacteria to avoid host defences and by acting as adhesins . Four such proteins of Staphylococcus aureus have been characterised: protein A (immunoglobulin binding protein), fibronectin binding proteins, collagen binding protein and the fibrinogen binding protein (clumping factor) . Site-specific mutants are being used to define their roles in pathogenesis in in vitro and in vivo models of adherence and infection. Med Clin (Barc), 1994 May 7, 102(17), 652 - 6 {Neurologic complications of infective endocarditis}; Almirante B et al.; BACKGROUND: Neurologic involvement is a frequent cause of morbidity and mortality in patients with endocarditis . The aim of this study was to evaluate the most relevant clinical, epidemiologic and evolutive characteristics of patients presenting neurologic complications during the evolution of endocarditis . METHODS: Fifty adult non intravenous drug addict patients who had neurologic complications during endocarditis were prospectively evaluated . The presence of cerebral ischemia, hemorrhage, or infectious complications were studied by established criteria . RESULTS: Neurologic complications were detected in 50 of the 282 patients (18%) with endocarditis diagnosed over 17 years in one institution . The most common complications were seen in patients with mitral endocarditis of either native valves (28%) or prosthetic valves (30%) . The prevalence was identical in the endocarditis of either type of valve (18%) . The most frequent complication was cerebral ischemia (29 episodes) and central nervous system infection (8 episodes) . Fifty-nine percent of the complications presented prior to the diagnosis of endocarditis with half of the remaining 41% occurring during the first 2 weeks of antibiotic treatment . Endocarditis by Staphylococcus aureus was associated with neurologic complications in 40% of the cases . Global mortality was 48%, being related with prosthetic endocarditis, the existence of cerebral hemorrhage or central nervous system infection, etiology by S . aureus and the presence of anticoagulant treatment . CONCLUSIONS: Neurologic complications are frequent during the evolution of infectious endocarditis, both or an initial feature and during its evolution . The presence of complications considerably impairs the prognosis of this disease. Gene, 1994 May 3, 142(1), 67 - 71 Photolabeling of the EcoP15 DNA methyltransferase with S-adenosyl-L-methionine; Ahmad I et al.; Radioactivity from S-adenosyl-L-{methyl-3H}methionine ({methyl-3H}AdoMet) was bound to the EcoP15 DNA methyltransferase (M.EcoP15) following short-wave ultraviolet (UV) irradiation . The labeled protein was subjected to polyacrylamide-gel electrophoresis in the presence of sodium dodecyl sulfate (SDS-PAGE), and detected by fluorography and autoradiography . Labeling was found to be dependent on the concentration of AdoMet and time of UV irradiation . The photolabeling by {methyl-3H}AdoMet was specific and blocked by S-adenosyl-L-homocysteine (AdoHcy) and sinefungin which are known to function as competitive inhibitors . Limited digestion of the M.EcoP15-AdoMet adduct by Staphylococcus aureus protease V8 generated three peptides of approx . 50, 32 and 30 kDa . Interestingly, only the 30-kDa peptide fragment contained radioactivity, as detected by SDS-PAGE, followed by fluorography and autoradiography . Further, sequencing of a few amino acids at the N-terminus of these peptides showed that the 30-kDa fragment was the N-terminal portion of M.EcoP15 . These results suggest that photolabeling is at the AdoMet-binding site and that the N-terminal half of M.EcoP15 may be involved in substrate binding. J Lab Clin Med, 1994 May, 123(5), 685 - 92 Early adhesion of bacteremic strains of Staphylococcus epidermidis to polystyrene: influence of hydrophobicity, slime production, plasma, albumin, fibrinogen, and fibronectin; Galliani S et al.; Twenty bacteremic strains of Staphylococcus epidermidis were characterized according to their hydrophobicity, their ability to produce slime, and their in vitro adhesion to polystyrene microtiter plates precoated or not with plasma proteins . Four strains of Staphylococcus aureus were also tested for adhesion . Slime production in S . epidermidis was not correlated with initial adhesion, whether measured qualitatively or by a quantitative method . Hydrophobicity (xylene:water partition) was well correlated with adhesion . Slime production, adhesion, and hydrophobicity were highly strain dependent among S . epidermidis organisms . For S . epidermidis, early adhesion was inhibited (10% to 98%) by albumin and fibronectin in all strains, by plasma (19 strains), and by fibrinogen (18 strains) . Stimulation occurred for one strain with plasma and two strains with fibrinogen . In contrast, adhesion was inhibited by albumin and markedly stimulated (twofold to 14-fold) by plasma, fibrinogen, and fibronectin for the four strains of S . aureus . Early adhesion of S . epidermidis to polymer surface appears to depend mainly on hydrophobicity and is usually impaired by plasma proteins, albumin, fibrinogen and fibronectin; with a heterogeneous behavior among the different strains tested . Slime production would interpose secondarily, after the first attachment. Clin Immunol Immunopathol, 1994 May, 71(2), 176 - 82 A single injection of Staphylococcus aureus enterotoxin B reduces autoimmunity in MRL/lpr mice; Gonzalo JA et al.; MRL/Mp-lpr/lpr (MRL/lpr) mice carry a mutation in the Fas gene whose product is involved in the regulation of lymphocyte apoptosis . This mutation is associated with the lpr phenomenon, i.e., a massive expansion of phenotypically abnormal CD4-CD8- cells ("double negative," DN) alpha/beta T cells (lpr cells) that becomes manifest at 3-4 months of age . As in normal mice, intravenous SEB injection into 2- or 6-month-old female MRL/lpr mice causes a transient expansion of SEB-reactive V beta 8+ T cells, followed by a deletion of this subset . In contrast, in the same animals, the frequency of abnormal V beta 8+CD4-CD8- cells is not modulated by SEB . Whereas DN T cells are completely resistant to SEB-mediated deletion in vivo, their precursors appear susceptible to SEB-induced deletion . Thus, a single injection of SEB prior to the surge of DN T cells in peripheral lymphoid organs, at 2 months of age, is sufficient to cause a stable long-term (6 months) deletion of DN cells . This is accompanied by a significant amelioration of autoimmune parameters (autoantibody titers, incidence of arthritis and nephritis), thus pointing to the feasibility of employing superantigens for simple manipulations of the immune repertoire that result in the long-term prophylaxis of autoimmune diseases. Arch Biochem Biophys, 1994 May 1, 310(2), 410 - 6 Mitochondrial hexokinase in brain: coexistence of forms differing in sensitivity to solubilization by glucose-6-phosphate on the same mitochondria; Kabir F et al.; Hexokinase (ATP:D-hexose 6-phosphotransferase, EC 2.7.1.1) is associated with mitochondria from brain of various species . The fraction of the bound activity that can be released in soluble form after incubation of the mitochondria with glucose-6-phosphate (Glc-6-P) has been shown to vary markedly among species (F . Kabir and J.E . Wilson, Arch . Biochem . Biophys . 300, 641-650, 1993) . A method has been developed by which mitochondria bearing significant amounts of bound hexokinase can be selectively immunoprecipitated by Staphylococcus aureus cells coated with anti-Type I hexokinase antibodies . Treatment of mitochondria from guinea pig, bovine, and human brain with Glc-6-P solubilized approximately 60, 40, and 20% of the bound hexokinase activity, respectively, but had no effect on the extent to which the mitochondria could be immunoprecipitated . This is consistent with the view that the residual hexokinase, resistant to solubilization with Glc-6-P, coexists on the same mitochondria bearing the Glc-6-P-sensitive form, i.e., removal of the latter does not prevent immunoprecipitation mediated by the Glc-6-P-resistant form . Mitochondrial porin has previously been shown to be involved in binding of hexokinase to mitochondria . Four porin species, having a common molecular weight but differing in isoelectric point, were detected, in the same relative amounts, in mitochondria from bovine and rat brain . The extent to which Glc-6-P solubilizes hexokinase from rat and bovine brain mitochondria is approximately 90 and approximately 40%, respectively . Thus the marked difference in sensitivity to solubilization with Glc-6-P cannot be attributed to differences in the relative amounts of different porin species present in rat and bovine mitochondria. J Med Microbiol, 1994 May, 40(5), 344 - 9 Coagulase deficiency in clinical isolates of Staphylococcus aureus involves both transcriptional and post-transcriptional defects; Vandenesch F et al.; The molecular basis of the non-expression of coagulase was investigated for 14 coagulase-negative isolates of Staphylococcus aureus obtained from different clinical samples . These isolates had typical S . aureus characteristics such as production of clumping factor, DNAase and protein A, but, with one exception, failed to produce detectable amounts of alpha-haemolysin . All 14 strains had DNA homologous to the coagulase gene (coa), but a coa-specific transcript was found in only seven of them . alpha-Haemolysin mRNA was detected in only eight strains without direct correlation to coa-mRNA expression . Thus, coagulase and alpha-haemolysin deficiencies in S . aureus may involve either transcriptional or post-transcriptional alterations although additional regulatory factors may influence the expression of both genes. J Fam Pract, 1994 May, 38(5), 473 - 7 The appropriateness of initial vancomycin dosing; Rodman DP et al.; BACKGROUND . Vancomycin use has markedly increased over the past several years because of an increased incidence of resistant organisms, particularly methicillin-resistant Staphylococcus aureus . Despite the availability of dosing nomograms and the use of peak and trough levels, vancomycin dosing has remained problematic . METHODS . All intravenous vancomycin orders over a 3-month period in a community and teaching hospital were screened for appropriateness of initial dosing based on available dosing nomograms . RESULTS . Of the 48 patients who received intravenous vancomycin, only 19 (39.6%) were given initial doses that achieved the desired serum concentration . There were no significant differences in the appropriateness of initial dosing between family medicine residents, attending physicians, and private staff physicians . Older patients in our study were at higher risk for overdosing, whereas younger patients were more likely to be underdosed . In this study, nomogram use could have yielded correct initial dosages in 40 of the 48 patients (83.3%) . CONCLUSIONS . Our study indicates a high percentage of inappropriate initial vancomycin dosing in a community and teaching hospital . The investigators believe inappropriate initial vancomycin dosing is common and may result in unnecessary expense, increased risk of therapeutic failures, and greater potential for adverse drug reactions . Increased use of vancomycin dosing nomograms could improve the rate of correct initial dosages. Am J Med Sci, 1994 May, 307(5), 340 - 1 Case report: fatal Staphylococcus aureus sepsis from single-donor platelet transfusion; Davda RK et al.; Bacterial sepsis is a rare and potentially fatal complication of platelet transfusions . Bacterial contamination is estimated to occur in 2% to 6% of platelet concentrates, but clinically apparent septicemia occurs in less than 1% of multi-donor platelet transfusions . The incidence of bacterial sepsis from single-donor platelet transfusions is significantly lower, possibly because of shorter storage time of apheresis platelets . The authors report a case of fatal Staphylococcus aureus sepsis from a single-donor platelet transfusion obtained through a closed system . They conclude that as the demand for platelet transfusion increases, recognition of bacterial sepsis from transfusions and continued reassessment of procedures for collection and storage of platelet concentrates is warranted. J Neurosurg, 1994 May, 80(5), 897 - 905 Blood-brain barrier permeability in staphylococcal cerebritis and early brain abscess; Lo WD et al.; The pattern of radiographic enhancement in cases of brain abscess has been extensively studied, but the magnitude of blood-brain barrier (BBB) damage that accompanies enhancement has not . The question of whether BBB permeability increases continuously as a cerebritis evolves into an abscess was studied . The tracers 3H-labeled aminoisobutyric acid and 14C-labeled butanol were used in a rat Staphylococcus aureus cerebritis model to measure simultaneously BBB permeability and blood flow . The rats were examined at 1, 2, 3, 5, or 7 days after inoculation, and tissue samples were collected from the cerebritis site and uninoculated regions . Permeability of the BBB in the cerebritis region increased to five times the normal values by 72 hours after inoculation, then reached a plateau . The plasma volume in the cerebritis region increased to six times greater than the normal value at 72 hours, then remained unchanged . Uninoculated brain in both ipsilateral and contralateral hemispheres showed no significant changes . Cerebral blood flow was not substantially altered at the inoculated or uninoculated sites . In this model, incidence of BBB damage rises rapidly, reaches a plateau, and does not continue to increase despite the ongoing evolution of a cerebritis into an abscess . The BBB damage is accompanied by an increase in the regional plasma volume, a novel finding that has not been previously reported in central nervous system inflammation . These results suggest that the vascular events contributing to brain edema formation become established early in the cerebritis phase and imply that control of the host's inflammatory response is important in the management of cerebritis-associated brain edema. J Infect Dis, 1994 May, 169(5), 1142 - 6 A superantigen as virulence factor in an acute bacterial infection; Rott O et al.; This study addresses the role of a bacterial superantigen as a potential virulence factor during an acute systemic infection . BALB/c mice were intravenously infected with a recombinant Staphylococcus aureus strain capable of producing plasmid-encoded staphylococcal enterotoxin B (SEB) or with the SEB plasmid-deficient parental strain . Infection with SEB-producing bacteria resulted in an initial expansion and subsequent decrease of circulating V beta 8+ T lymphocytes . This numeric decrease was accompanied by a SEB-specific state of hyporesponsiveness of splenic T cells . In parallel with SEB-triggered unresponsiveness of a large proportion of T lymphocytes, a weakening of the overall T cell responsiveness towards the invading bacteria was found . Furthermore, the production of SEB altered the kinetics of bacterial clearance: Animals infected with the SEB-producing variant showed a significantly elevated bacterial burden and could less efficiently clear the bacteria . However, the overall effect of SEB production on the course of bacterial infection was surprisingly weak, suggesting that the superantigen was only a minor virulence factor for the bacterium. J Bacteriol, 1994 May, 176(9), 2751 - 3 Serum inhibits penicillin-induced L-form growth in Staphylococcus aureus: a note of caution on the use of serum in cultivation of bacterial L-forms; Shimokawa O et al.; Heat-inactivated horse serum inhibited penicillin-induced L-form colony formation in Staphylococcus aureus when included in an osmotically stabilized culture medium . Most, perhaps all, L-form colonies that appeared with low frequencies on the serum-supplemented medium were of the penicillin-independent, stable type . This relationship must be taken into account when use of serum is considered for L-form cultivation. Infect Immun, 1994 May, 62(5), 1843 - 7 Site-directed mutagenesis of the alpha-toxin gene of Staphylococcus aureus: role of histidines in toxin activity in vitro and in a murine model; Menzies BE et al.; Staphylococcus aureus alpha-toxin is a membrane-damaging exoprotein that oligomerizes to form transmembrane pores . Chemical modification of histidines with diethylpyrocarbonate has been shown to reduce the hemolytic activity of alpha-toxin, suggesting that one or more of the histidine residues is important for toxin function . To individually assess the functional importance of each of the four histidine residues (residues 35, 48, 144, and 259), we used oligonucleotide-directed mutagenesis of the cloned alpha-toxin gene to replace each histidine with leucine . The mutant toxins were expressed in S . aureus and evaluated for hemolytic activity in vitro and for lethality in an intraperitoneal murine model . Substitution of histidine 35 with leucine produced a mutant toxin (H35L) without hemolytic or lethal activity . Mutant toxins H48L, H144L, and H259L exhibited 7, 16, and 46%, respectively, of the hemolytic activity of wild-type toxin . Immunoblotting of purified H35L toxin incubated with liposomal membranes demonstrated intact membrane binding and hexamer formation that was clearly detectable but reduced compared with that of the wild-type toxin . This suggests that hexamer formation alone is not sufficient for the expression of alpha-toxin activity . The nature of the defect underlying the lack of activity of the H35L mutant toxin remains to be elucidated but may involve failure of the hexamer to span the lipid bilayer to form a transmembrane pore or a change in the internal surface and permeability characteristics of the pore. Infect Immun, 1994 May, 62(5), 1719 - 25 Role of the sar locus of Staphylococcus aureus in induction of endocarditis in rabbits; Cheung AL et al.; A regulatory locus on the Staphylococcus aureus chromosome, designated sar, is involved in the expression of cell wall proteins, some of which are potentially important in the pathogenesis of endocarditis . For instance, mutant 11D2 (sar::Tn917LTV1) was found to bind substantially less to matrix proteins (i.e., fibrinogen and fibronectin) than parent strain DB . Remarkably, these two strains did not differ in other phenotypes considered important in the initiation of endocarditis (e.g., binding to platelets and resistance to platelet-derived microbicidal proteins) . The isogenic pair were compared for pathogenicity in a rabbit endocarditis model . There were significant differences in infectivity rates between the two strains (71 and 88% for DB versus 17 and 42% for mutant 11D2 at inocula of 10(3) and 10(4) CFU, respectively) . In early adherence studies, parent DB adhered substantially better than the mutant to valvular vegetations at an inoculum of 10(6) CFU (P = 0.05) . Southern blot analysis of colonies indicated that the location of the Tn917LTV1 insert in mutant 11D2 remained stable after animal passage . In vitro adherence assays revealed that mutant 11D2 was less adherent to cultured human endothelium than parent DB . These studies suggest that the sar locus is involved in the initial adherence of S . aureus to the fibrin-platelet-endothelium matrix on damaged valvular endothelium. Blood, 1994 May 1, 83(9), 2646 - 53 Monocyte dysfunction in patients with Gaucher disease: evidence for interference of glucocerebroside with superoxide generation; Liel Y et al.; Gaucher disease patients are occasionally affected by chronic or fulminant infections . Since Gaucher cells originate from tissue phagocytes, we studied the functional implications of glucocerbroside accumulation on phagocytes in Gaucher disease patients . Circulating monocytes and granulocytes from nine type I Gaucher disease patients, and matched controls, were studied . Evaluation of phagocytic activity included (1) maximal superoxide generation rates following stimulation by phorbol 12-myristate 13-acetate (PMA), opsonized zymosan (OZ), or formyl-methionyl-leucylphenylalanine (FMLP); (2) nitroblue tetrazolium reduction test (NBT); (3) chemotaxis toward FMLP; (4) phagocytosis of OZ particles; and (5) killing activity against Staphylococcus aureus . Superoxide generation in monocytes following PMA, OZ, and FMLP stimulation was significantly suppressed at 52% +/- 15%, 39% +/- 8%, and 51% +/- 11% of control, respectively . Superoxide generation in granulocytes was normal . NBT reduction, staphylococcal killing, and phagocytosis were also markedly decreased in monocytes, and normal in granulocytes . Mean chemotaxis rates were normal in both monocytes and granulocytes; however, decreased chemotactic rates were observed in some patients . The abnormality of superoxide generation could be reproduced in a dose- and time-dependent manner in normal circulating monocytes incubated with glucocerebroside . Superoxide generation in glucocerebroside-conditioned normal monocytes in a cell-free system showed normal superoxide generation, reflecting the integrity of the NADPH oxidase complex itself . These results demonstrate markedly compromised phagocytic functions in circulating monocytes in Gaucher disease patients . These abnormalities can be attributed to accumulation of glucocerebroside, since it could be reproduced in normal monocytes incubated with glucocerebroside . Similar abnormalities in Gaucher cells throughout the reticuloendothelial system could impair host defense, and may be of particular importance in the pathogenesis of osteomyelitis in Gaucher disease patients. J Neurochem, 1994 May, 62(5), 1795 - 803 Sequestration of muscarinic cholinergic receptors in permeabilized neuroblastoma cells; Slowiejko DM et al.; The feasibility of using a permeabilized preparation of human SH-SY-5Y neuroblastoma cells for studies of muscarinic acetylcholine receptor (mAChR) sequestration has been evaluated . Exposure of cells permeabilized with digitonin, streptolysin-O, or the alpha-toxin from Staphylococcus aureus to oxotremorine-M (Oxo-M) for 30 min resulted in a 25-30% reduction in the number of cell surface mAChRs, as monitored by the loss of N{3H}methylscopolamine ({3H}NMS) binding sites . The corresponding value for intact cells was 40% . For cells permeabilized with 20 microM digitonin, the Oxo-M-mediated reduction in {3H}NMS binding was time (t1/2 approximately 5 min) and concentration (EC50 approximately 10 microM) dependent and was agonist specific (Oxo-M > bethanechol = arecoline = pilocarpine) . In contrast, no reduction in total mAChR number, as monitored by the binding of {3H}quinuclidinyl benzilate, occurred following Oxo-M treatment . The loss of {3H}NMS sites observed in the presence of Oxo-M was unaffected by omission of either ATP or Ca2+, both of which are required for stimulated phosphoinositide hydrolysis, but could be inhibited by the inclusion of guanosine 5'-O-(2-thiodiphosphate) . mAChRs sequestered in response to Oxo-M addition were unmasked when the cells were permeabilized in the presence of higher concentrations of digitonin (80 microM).(ABSTRACT TRUNCATED AT 250 WORDS) J Immunol, 1994 May 1, 152(9), 4604 - 11 Fc gamma receptor cross-linking down-regulates IL-1 receptor antagonist and induces IL-1 beta in mononuclear phagocytes stimulated with endotoxin or Staphylococcus aureus; Marsh CB et al.; The cross-linking of monocyte Fc gamma R is a potent stimulus for IL-1ra production but does not induce IL-1 beta . However, during systemic infection, IgG-coated bacteria can activate mononuclear phagocytes via both cell wall components and opsonized IgG . Therefore, we analyzed the effect of combinations of Fc gamma R cross-linking and bacterial cell wall components on mononuclear phagocyte IL-1 beta and IL-1ra production . Human mononuclear cells and monocytes were cultured either alone or with combinations of immobilized IgG, LPS, or heat-killed Staphylococcus aureus (HKSA) . Cells cultured on immobilized IgG released large amounts of IL-1ra but no detectable IL-1 beta . In response to LPS, mononuclear cells released IL-1ra at 1000-fold lower doses of LPS than was required to induce IL-1 beta . However, when measured in the presence of immobilized IgG, the LPS sensitivity for IL-1 beta release increased 100-fold, whereas IL-1ra release correlated inversely with the LPS dose . Furthermore, HKSA, a nonendotoxin stimulus, affected mononuclear cell IL-1 beta and IL-1ra release similarly . In addition, polymyxin B, a specific endotoxin inhibitor, blocked the LPS, but not the HKSA-induced changes in IL-1 beta and IL-1ra secretion, irrespective of immobilized IgG co-stimulation . In summary, these results suggest that mononuclear phagocyte stimulation with immobilized IgG favors IL-1ra over IL-1 beta production . Conversely, the addition of LPS or HKSA to the Fc gamma R-stimulated cells augments IL-1 beta but suppresses IL-1ra production. J Antimicrob Chemother, 1994 May, 33(5), 959 - 67 Microbiological tests to predict treatment outcome in experimental device-related infections due to Staphylococcus aureus; Zimmerli W et al.; Treatment outcome of experimental device-related infections cannot be predicted by the results of standard susceptibility tests such as MIC . Microorganisms involved in such infections have a slow growth rate and adhere to surfaces . Therefore, laboratory tests were developed taking into account these properties and compared with the treatment outcome in an animal model . Vancomycin, teicoplanin, ciprofloxacin and fleroxacin were tested alone, or in combination with rifampicin for their ability to cure experimental device-related infections in guinea pigs due to Staphylococcus aureus ATCC 29213 . Rifampicin alone or in combination was significantly more effective than the other four drugs (P < 0.001) . Combined treatment with rifampicin had a higher cure rate than rifampicin alone . Treatment success was not predicted by an antibiotic trough level exceeding the MIC at site of infection . In contrast, drug efficacy was predicted if the stationary-phase MBC was in the sensitive range, and if glass-adherent S . aureus was killed by low drug concentrations. Int Immunol, 1994 May, 6(5), 739 - 49 Expression of a G-protein beta subunit-related gene during lymphocyte activation; Shan X et al.; Using a subtractive strategy, we have cloned an activation-related gene from a human B cell cDNA library . Sequence analysis revealed that this gene was identical to H12.3, a gene belonging to an expanding family of guanine nucleotide-binding protein beta subunits . The expression of H12.3 was inducible in the late phase of mitogen-stimulated T and B cells . In T cells, IL-2 and IL-4 by themselves had no direct effect on the expression of H12.3, but they could augment the level of steady-state H12.3 mRNA stimulated by phytohemagglutinin . On the other hand, IFN-gamma and IL-6 had no obvious effect on the expression in B cells with or without Staphylococcus aureus Cowan I-stimulation . Cyclosporin A, a strong immunosuppressant, inhibited the mitogen-stimulated expression of H12.3, but rapamycin, another such agent, did not . In synchronized Jurkat cells, the expression of H12.3 had no cell cycle-dependent decrease in S and G2/M phase, while cyclin E, which controls the progression of the cell cycle in late G1 phase, did show a periodic expression pattern . The results suggest that H12.3 might be involved in regulation of lymphocyte activation. Clin Infect Dis, 1994 May, 18(5), 802 - 4 Liver abscesses due to Staphylococcus aureus in a patient with AIDS who underwent small bowel biopsy: case report and review; Gunnarsson G et al.; A 36-year-old man with overgrowth of Staphylococcus aureus in the small bowel underwent a biopsy of the small bowel and presented 5 weeks later with multiple liver abscesses . To our knowledge, small bowel biopsy has never been associated with liver abscesses . We discuss the literature on liver abscesses in patients with AIDS and the risks of bacteremia following upper endoscopy and small bowel biopsy and propose that the etiology of the liver abscesses in our patient was seeding of the portal vein following biopsy of a small bowel that had significant bacterial overgrowth. Clin Infect Dis, 1994 May, 18(5), 750 - 4 Infectious intracranial complications of sinusitis, other than meningitis, in children: 12-year review; Rosenfeld EA et al.; Sinusitis is usually a mild illness in children, but intracranial complications can be life-threatening . We retrospectively reviewed nine cases of intracranial infections secondary to paranasal sinusitis that occurred over a 12-year period, excluding patients with orbital infection only . Cases were highly age- and sex-associated: the median age was 14 years, 89% of patients were > 9 years of age, and seven (78%) of the nine patients were male . Symptoms included fever (67%), headache (67%), eye swelling (56%), and seizure (33%) . Rhinorrhea was uncommon (22%) . Only two patients (22%) had had previous episodes of sinusitis . Staphylococcus aureus and anaerobes were the predominant intracranial isolates . Computed tomography scans of the head showed progression of disease in patients treated with antibiotics alone; surgical drainage was required for all patients . The duration of therapy after surgery was 3-8 weeks . Only one patient (11%) had persistent neurological sequelae . We conclude that (1) teenage males are at greatest risk of developing intracranial infections from sinusitis, (2) common symptoms of sinusitis such as rhinorrhea may not always occur, and (3) outcome can be excellent when a combined medical/surgical approach is used for therapy. Protein Eng, 1994 May, 7(5), 655 - 62 A pore-forming protein with a metal-actuated switch; Walker B et al.; Staphylococcal alpha-hemolysin, a pore-forming exotoxin, is a polypeptide of 293 amino acids that is secreted by Staphylococcus aureus as a water-soluble monomer . It assembles to form hexameric pores in lipid bilayers . Previous studies of pore formation have established the involvement of a central glycine-rich loop . Here, we show that when five consecutive histidine residues replace amino acids 130-134 at the midpoint of the loop, they provide a switch with which pore activity can be (i) turned off by micromolar concentrations of divalent zinc ions and (ii) turned back on with the chelating agent EDTA . Planar bilayer recordings show that Zn2+ and EDTA can act on open channels from either side of the bilayer and thus demonstrate that the central loop lines part of the conductive pathway . Our results suggest that genetically-engineered pore-forming proteins might make useful components of metal ion sensors. Eur J Clin Microbiol Infect Dis, 1994 May, 13(5), 420 - 4 Comparison of four genotyping assays for epidemiological study of methicillin-resistant Staphylococcus aureus; van Belkum A et al.; Twenty-six methicillin-resistant Staphylococcus aureus strains were genetically differentiated by interrepeat PCR and the results compared with those of ribotyping, pulsed-field gel electrophoresis (PFGE) and random amplified polymorphic DNA analysis obtained in a previous study for the same strains . The comparison showed that the PCR-mediated assays were as discriminatory as PFGE, whereas ribotyping was the least powerful genotyping method . Due to the ease of performance, PCR fingerprinting may become the method of choice for establishing clonal relationship among Staphylococcus aureus isolates. Antimicrob Agents Chemother, 1994 May, 38(5), 945 - 52 Synthesis and antimicrobial activity of dimethyl- and trimethyl-substituted phosphonium salts with alkyl chains of various lengths; Kanazawa A et al.; Various phosphonium salts possessing single or double alkyl chains of various lengths (C10 to C18) were prepared as cationic biocides, and their antimicrobial activities against 11 typical strains of microorganisms including methicillin-resistant Staphylococcus aureus (MRSA) were evaluated . The phosphonium salts with long alkyl chains were found to show high levels of antimicrobial activity . Their activities depended strongly on the molecular structure, and a correlation between antimicrobial activity and molecular structure was observed . In the alkyltrimethylphosphonium salts, the bactericidal activity against S . aureus and Escherichia coli increased with increasing alkyl chain length, and the compound with the longest alkyl chain (C18) killed all the bacterial cells (ca . 10(7) cells per ml) within 30 min of contact at concentrations of 2.8 and 28 microM, respectively . In contrast, the bactericidal activity of dialkyldimethylphosphonium salts was found to decrease as the chain length of the substituents increased . It is significant that the phosphonium biocide containing double decyl groups exhibited the broadest spectrum of activity against microorganis