J Ocul Pharmacol Ther, 2000 Feb, 16(1), 75 - 9 Antibiotic treatment of orbital cellulitis: an analysis of pathogenic bacteria and bacterial susceptibility; Chang CH et al.; The proper choice of effective antibiotics is a mainstay for the treatment of orbital cellulitis . The lack of native data regarding the microorganism causing the infection and its antibiotic sensitivity prompted us to conduct this study . We retrospectively collected 29 cases of orbital cellulitis admitted to Chung-Ho Memorial Hospital of Kaohsiung Medical College from January 1994 to September 1998 . The effectiveness of antibiotics with bacterial susceptibility was analyzed . Of the 29 cases, fifteen were male and fourteen female . The patients ranged in age from 7 months to 79 years (mean, 37.6 years) . Sinusitis (9 cases, 31.0%) is the most common etiology . Fourteen cases received both medical and surgical treatments . Eighteen cases had purulent discharge from the infection areas sent for culture isolation of the microorganism . The culture positive rate was 50% (9 in 18 cases) . The Staphylococcus aureus (5 cases) was the most common pathogen . The bacterial susceptibility test showed drug resistance of 100% for penicillin G (seven out of seven cases; 7/7), 100% for ampicillin (10/10), and 0% for amikacin (0/3) and vancomycin (0/7) . Penicillin and ampicillin are not effective for those isolated bacteria . Oxacillin and gentamicin, frequently used in first line treatment, might encounter drug resistance in some cases . Amikacin and vancomycin, without any resistance in bacterial susceptibility tests, could be used in vision-threatening, critical, and intractable cases. J Biomater Sci Polym Ed, 1999, 10(12), 1207 - 21 Inhibition by heparin and derivatized dextrans of Staphylococcus epidermidis adhesion to in vitro fibronectin-coated or explanted polymer surfaces; Francois P et al.; The ability of Staphylococcus aureus to recognize several extracellular matrix or plasma proteins (e.g., fibrinogen, fibronectin, and collagen) promotes bacterial attachment to artificial surfaces . Whereas most S . aureus clinical isolates elaborate a wide repertoire of bacterial surface receptors' called adhesins, exhibiting specific binding of individual host proteins, S . epidermidis is lacking most of such protein adhesins . To document the interactions between S . epidermidis and various surface-adsorbed proteins, we first compared promotion of bacterial attachment by seven purified human proteins immobilized onto poly(methyl methacrylate) (PMMA) coverslips . Only two of them, namely fibronectin and fibrinogen, exhibited adhesion-promoting activities . In the presence of native heparin or two functionalized dextrans (CMDBS for Carboxy Methyl, Benzylamide sulfonate/sulfate), a dose-dependent inhibition of S . epidermidis adhesion to fibronectin-coated, but not to fibrinogen-coated surfaces was observed . The inhibitory effects of each CMDBS were much stronger than that of native heparin . In contrast, a control highly negatively charged, dextran exclusively substituted with carboxy methyl groups exerted no inhibition on S . epidermidis adhesion . To evaluate how CMDBS could interfere with S . epidermidis attachment to coverslips coated in vivo with extracellular matrix components, we also tested PMMA surfaces retrieved from tissue cages subcutaneously implanted in guinea pigs . Each CMDBS, but not heparin, strongly inhibited S . epidermidis adhesion to explanted coverslips, even in the presence of tissue cage fluid . In conclusion, fibronectin plays an important role in promoting S . epidermidis attachment to implanted biomaterials . Furthermore, S . epidermidis adhesion to fibronectin-coated or implanted biomaterials can be efficiently blocked in vitro by CMDBS. J Biol Chem, 2000 Feb 18, 275(7), 4654 - 9 Inhibition of the Staphylococcus aureus NADPH-dependent enoyl-acyl carrier protein reductase by triclosan and hexachlorophene; Heath RJ et al.; Enoyl-acyl carrier protein reductase (FabI) plays a determinant role in completing cycles of elongation in type II fatty acid synthase systems and is an important target for antibacterial drugs . The FabI component of Staphylococcus aureus (saFabI) was identified, and its properties were compared with Escherichia coli FabI (ecFabI) . ecFabI and saFabI had similar specific activities, and saFabI expression complemented the E . coli fabI(Ts) mutant, illustrating that the Gram-positive FabI was interchangeable with the Gram-negative FabI enzyme . However, ecFabI was specific for NADH, whereas saFabI exhibited specific and positive cooperative binding of NADPH . Triclosan and hexachlorophene inhibited both ecFabI and saFabI . The triclosan-resistant ecFabI(G93V) protein was also refractory to hexachlorophene inhibition, illustrating that both drugs bind at the FabI active site . Both the introduction of a plasmid expressing the safabI gene or a missense mutation in the chromosomal safabI gene led to triclosan resistance in S . aureus; however, these strains did not exhibit cross-resistance to hexachlorophene . The replacement of the ether linkage in triclosan by a carbon bridge in hexachlorophene prevented the formation of a stable FabI-NAD(P)(+)-drug ternary complex . Thus, the formation of this ternary complex is a key determinant of the antibacterial activity of FabI inhibitors. Clin Infect Dis, 2000 Feb, 30(2), 368 - 73 Pathogenic significance of methicillin resistance for patients with Staphylococcus aureus bacteremia; Soriano A et al.; To assess whether methicillin resistance is a microbial characteristic associated with deleterious clinical outcome, we performed a cohort study on 908 consecutive episodes of Staphylococcus aureus bacteremia and a case-control study involving 163 pairs of patients matched for preexisting comorbidities, prognosis of the underlying disease, length of hospitalization, and age . Of 908 bacteremic episodes, 225 (24.8%) were due to methicillin-resistant S . aureus (MRSA) . Multivariate analysis did not reveal that methicillin resistance was an independent predictor for mortality when shock, source of bacteremia, presence of an ultimately or rapidly fatal underlying disease, acquisition of the infection in an intensive care unit (ICU), inappropriate empirical therapy, female sex, and age were taken into account . Nonetheless, methicillin resistance was an independent predictor for shock . The case-control study could not confirm that shock was linked to MRSA when prior antimicrobial therapy, inappropriate treatment, ICU residence, and female sex were considered . Our data suggest that cohort studies tend to magnify the relationship of MRSA with clinical markers of microbial pathogenicity and that this effect is a shortcoming of these kind of studies that is caused by inadequate control for underlying diseases. Clin Infect Dis, 2000 Feb, 30(2), 322 - 7 Methicillin-resistant Staphylococcus aureus: the other emerging resistant gram-positive coccus among liver transplant recipients; Singh N et al.; We undertook a study of the characteristics and clinical impact of infections due to methicillin-resistant Staphylococcus aureus (MRSA) after liver transplantation . Of 165 patients who received liver transplants at our institution from 1990 through 1998, 38 (23%) developed MRSA infections . The predominant sources of infection were vascular catheters (39%; n=15), wound (18%; n=7), abdomen (18%; n=7), and lung (13%; n=5) . A significant increase in MRSA infections (as a percentage of transplant patients infected per year) occurred over time (P=.0001) . This increase was greater among intensive care unit patients (P=.001) than among nonintensive care unit hospital patients (P=.17) . Cytomegalovirus seronegativity (P=.01) and primary cytomegalovirus infection were significantly associated with MRSA infections (P=.005) . Thirty-day mortality among patients with MRSA infections was 21% (8/38) . Mortality was 86% in patients with bacteremic MRSA pneumonia or abdominal infection and 6% in those with catheter-related bacteremia (P=.004) . Thus the incidence of MRSA infection has increased exponentially among our liver transplant recipients since 1990 . These infections have unique risk factors, time of onset, and a significant difference in site-specific mortality; deep-seated bacteremic infections, in particular, portend a grave outcome. J Med Microbiol, 2000 Feb, 49(2), 187 - 92 Spread of the Brazilian epidemic clone of a multiresistant MRSA in two cities in Argentina; Da Silva Coimbra MV et al.; Methicillin-resistant Staphylococcus aureus (MRSA) is recognised as an important cause of nosocomial infection . The spread of some MRSA epidemic clones is well documented . In Brazil, and more recently in Portugal, a considerable number of hospital infections has been caused by a unique multiresistant MRSA clone designated as the Brazilian epidemic clone . This paper describes the spread of this clone in hospitals in two cities in Argentina. ASAIO J, 2000 Jan-Feb, 46(1), 33 - 7 Bacterial components inhibit fibroblast proliferation in vitro; Edds EM et al.; Perigraft fluid from Staphylococcus epidermidis infected grafts in a mouse model significantly inhibits fibroblast proliferation (60-98% at 7 and 28 days), compared with perigraft fluid from sterile grafts . The fibroblast inhibitor was trypsin-heat resistant and dependent primarily upon the bacteria, not the host proinflammatory mediators or the vascular graft biomaterial . We tested the inhibitory properties of S . epidermidis strains RP62A (slime producer) and RP62NA (nonslime producer) and Staphylococcus aureus strain 502a, using an in vitro tritiated thymidine murine fibroblast (ATCC CCL-12) proliferation assay . Whole killed bacteria, disrupted bacteria (live and killed), bacterial supernatants, and purified cell wall products (peptidoglycan, teichoic acid, and lipoteichoic acid from disrupted bacteria) were studied . Significant fibroblast inhibition occurred for all three bacterial strains with disrupted bacteria (live or killed) and cell free bacteria derived supernatants . The fibroblast inhibitor from disrupted slime producing S . epidermidis was trypsin-heat resistant . The fibroblast inhibitor from disrupted S . aureus and supernatants for all three bacterial strains at 1 x 10(7) were trypsin-heat sensitive . Fibroblast inhibition was not dependent upon bacterial viability and not mediated by bacterial cell wall products . In conclusion, components of slime and nonslime producing S . epidermidis and S . aureus inhibit fibroblast proliferation. Vet Microbiol, 2000 Jan, 71(1-2), 89 - 101 The dynamics of Staphylococcus aureus intramammary infection in nine Danish dairy herds; Larsen HD et al.; The aim of the present study was to examine the diversity of Staphylococcus aureus isolates from bovine intramammary infections (IMI) in nine dairy herds, and compare these with isolates from other sites on the cows by phage- and ribotyping . Whether colonisation of milkers with S . aureus could be a source of infection for bovine IMI was investigated . In addition, 100 epidemiologically unrelated S . aureus isolates from asymptomatic human carriers were also phage- and ribotyped to compare the human and bovine reservoir of S . aureus in Denmark . A total of 625 S . aureus isolates from bovine IMI, bovine skin lesions, milking personnel, and non-farm-related human carriers were included in the study . Certain types predominated in one or several herds during the study period of one-and-a-half to two years, whereas the presence of other types was of a more sporadic nature . Within the individual herds, there was a close correspondence between ribo- and phage types of S . aureus isolated from bovine intramammary infections and skin lesions . Isolates from milking personnel, however, were not identical to any of the predominant intramammary strains . Furthermore, several of the isolates from milking personnel showed ribo- and phage patterns identical to S . aureus isolates from human carriers . The findings of the present study underline the importance of strict milking hygiene and improvement of current mastitis therapy . The results support the hypothesis that some S . aureus mastitis strains are more contagious, virulent or persistent than others . The human reservoir of S . aureus does not play a major role as a source of bovine intramammary infections. Vet Microbiol, 2000 Jan, 71(1-2), 53 - 8 Phylogenetic relationships of Staphylococcus aureus from bovine mastitis based on coagulase gene polymorphism; Su C et al.; Coagulase gene restriction fragment length polymorphism (RFLP) patterns were analyzed to determine the phylogenetic relationship among isolates of Staphylococcus aureus from the Czech Republic (n = 27), France (n = 48), Korea (n = 115) and the United States (n = 278) . A total of 468 isolates of S . aureus were subtyped into 41 coagulase genotypes . Cluster analysis placed the 41 types into nine clusters . Eighteen API Staph profiles were determined for 102 S . aureus isolates representing 1 to 4 isolates of each coagulase type . The results of the study suggest that based on coagulase gene RFLP analysis, several genetic variants of S . aureus are prevalent . Comparison of coagulase and API Staph profiles indicated that the two identification system were independent of each other. Lett Appl Microbiol, 1999 Nov, 29(5), 347 - 9 Production of enterotoxins and toxic shock syndrome toxin by Staphylococcus aureus isolated from bovine mastitis in Brazil; Cardoso HF et al.; The production of toxic shock syndrome toxin 1 (TSST-1) and enterotoxins (SE) A, B, C and D by bovine mastitis isolates of Staphylococcus aureus was evaluated by immunodiffusion using the Optimum-Sensitivity Plate method . S . aureus strains were isolated from bovine mastitis in 23 dairy herds in the state of Minas Gerais, Brazil, during 1994-9 . Of 127 isolates, 83 (65.04%) produced one or several toxins, and among them production of SE was found in 54 (43.0%) isolates, of which 1138 (29.09%) secreted enterotoxin identified as type D . TSST-1 was found in 5829 (45.723.0%) isolates. Biotechnol Prog, 2000 Jan-Feb, 16(1), 116 - 24 A reusable and specific protein A-coated piezoelectric biosensor for flow injection immunoassay; Lu HC et al.; A hydrophilic matrix of periodate-oxidized dextran was used as a double-sided linker to covalently immobilize Staphylococcus aureus protein A (SpA) molecules onto a poly-L-lysine-modified piezoelectric crystal surface to improve their stability, activity, and binding specificity with human immunoglobulin G (IgG) in flow injection assays . The prepared sensing crystals displayed best sensitivity and reusability at a flow rate of 140 microL/min . A human IgG concentration as low as 0.3 nM can be detected by this system . Up to 19 successive assay repetitions were achieved without significant loss of sensitivity using the same crystal . The analysis of adsorption kinetics indicates that such a preparation can greatly increase the amount of available active human IgG binding sites on immobilized SpA . Hardly any response arising from unspecific binding was detected . In addition, the sensing crystal prepared by this method was found to retain activity better than one prepared via direct deposition when stored in either wet or dry states . Finally, the prepared SpA-coated crystals were applied to the affinity immobilization of polyclonal goat anti-Schistosoma japonicum glutathione-S-transferase (GST) and were able to subsequently detect GST and its genetically engineered mutant either in a purified form or in the crude cell lysate. J Antimicrob Chemother, 2000 Feb, 45(2), 139 - 44 MecI represses synthesis from the beta-lactamase operon of Staphylococcus aureus; Lewis RA et al.; Plasmid diploids were constructed in Staphylococcus aureus to study the effect of the repressor of methicillin resistance (MecI) on the synthesis of both beta-lactamase and the beta-lactamase repressor (BlaI) . MecI-mediated repression of the synthesis of beta-lactamase was shown by reduction in the specific activity of nitrocefinase in bacteria containing a plasmid carrying mecI but not when containing the same plasmid deleted for mecI . Antisera prepared against purified MecI and against purified BlaI were used in Western blots which showed that MecI repressed the synthesis of BlaI in these diploids . The interactions between the mec operon and the bla operon are discussed. Hinyokika Kiyo, 1999 Dec, 45(12), 835 - 7 {Giant psoas abscess with aggressive extension: report of a case}; Iwaki H et al.; We report a case of giant psoas abscess with aggressive extension outside the muscles of the iliopsoas component . A 57-year-old man was admitted to our hospital, presenting with right flank pain and severe general malaise . He had been diabetic, but no treatment had been performed for diabetes . Leukocytosis, positive CRP and hyperglycemia were noted, but he was nearly afebrile on admission . Computerized tomography revealed a large multilocular mass in the right retroperitoneal space involving the ipsilateral psoas muscle . The diagnosis was not apparent until the 12th hospital day, when moderate grade fever was noted and brownish purulent fluid was obtained by percutaneous puncture of the mass . Staphylococcus aureus was isolated on culture . Antibiotic chemotherapy was started, and ultrasound-guided percutaneous drainage was then performed under the diagnosis of psoas abscess . At that time, the abscess was aggressively extending from the iliopsoas component into the pelvic floor, involving the rectus muscle, the gluteal muscles and formation of subucutaneous lesions . At 46 days after drainage, surgical resection of the abscess with removal of the adjucent tissue was performed because of persistent discharge of pus and multiple residual lesions . The postoperative course was uneventful, and there has been no recurrence . Many cases of psoas abscess have been reported in the Japanese literature . Prompt drainage, either percutaneously or surgically are required . Surgical resection of the abscess, with not only opening the cavity but also removal of the adjacent tissue, may be recommended in some cases, especially those diffuse or multilocular lesions. J Hosp Infect, 1999 Dec, 43(4), 281 - 91 Antibiotic and biocide resistance in methicillin-resistant Staphylococcus aureus and vancomycin-resistant enterococcus; Suller MT et al.; Concern has been growing regarding the potential of antibiotic and disinfectant co-resistance in clinically important bacteria . In this study, the susceptibilities of methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-sensitive Staphylococcus aureus (MSSA) to chlorhexidine (CHX), the quaternary ammonium compounds cetylpyridinium chloride (CPC) and benzalkonium chloride (BC), triclosan, dibromopropamidine isethionate (DBPI) and triclocarban were compared . MRSA exhibited low-level resistance to CHX and the QACs, with MICs of 1.5 to 3-fold (CHX), and 2 to 4-fold (QACs) higher than MSSA . However, the MIC values for MRSA ranged between 0.025 (the MIC of MSSA) and 1 microg/mL with triclosan, and between <5 (the MIC of MSSA) and 75 microg/mL with DPBI . Nevertheless, these strains remain relatively sensitive to most of these antimicrobial agents . The bactericidal efficacy of CHX, CPC and DBPI (with the exception of one strain) correlated with their MIC value . This was not observed using triclosan; MRSA and MSSA strains were equally susceptible to its killing effect, regardless of MIC . The permeabilizing agent, ethylenediamine tetraacetic acid (EDTA) was unable to potentiate the antibacterial activities of the biocides against any of the strains tested . Attempts to select for staphylococcal strains with increased resistance to triclosan, CPC or CHX, using disc diffusion, step-wise broth, or repeated exposure/recovery technique, were only partially successful, and resistance was found to be unstable . The susceptibilities of vancomycin-resistant enterococcus (VRE) and vancomycin-sensitive enterococcus (VSE) to the biocides were also compared and found to be similar both in terms of MIC testing and time-kill studies. J Hosp Infect, 1999 Dec, 43 Suppl, S269 - 74 Antibiotic policies in Central/Eastern Europe (CEE) after 1990; Krcmery V et al.; Significant political and social changes in Eastern Europe have caused dramatic changes in healthcare: Centralized state drug policies were decentralized, vaccination strategies in some countries changed and directed financing of healthcare replaced by one or multiple health management organisations (HMO) . Centralized Infectious Diseases and/or Antibiotic Resistance surveillance were stopped after 1990 in four of six countries but resurrected after 1996 in Hungary, Poland and Slovakia . According to antibiotic (ATB) resistance and nosocomial infection rates, there are some differences in comparison to Western and Northern Europe e.g., a higher incidence of penicillin resistant pneumococci, ampicillin resistant H . influenzae and methicillin resistant Staphylococcus aureus . Because of increasing consumption of ATB before 1991, due to the free market, pharmaceutical marketing and reimbursement policies, several strategies to decrease consumption and/or resistance were implemented such as restriction of outpatient use, national and hospital formularies and Health Management Organizations-based restrictions . Probably due to the short time scale, no significant reduction in resistance has been documented although antibiotic consumption has declined. J Hosp Infect, 1999 Dec, 43 Suppl, S3 - 7 Implications of vancomycin-resistant Staphylococcus aureus; Tenover FC; Strains of Staphylococcus aureus with reduced susceptibility to glycopeptides have been reported from Japan (multiple strains), the United States (four strains), and Europe (France, the UK and Spain) and the Far East (Hong Kong and Korea) . The isolates from the US, France, and strain Mu50 from Japan, demonstrate vancomycin MICs of 8 microg/mL by broth microdilution testing and appear to have developed from pre-existing methicillin-resistant S . aureus (MRSA) infections . The strain from the UK and other parts of Europe appears heteroresistant to vancomycin and has MIICs in the 1-2 microg/mL range . Many of the isolates with reduced susceptibility to glycopeptides have been associated with therapeutic failures with vancomycin . Although nosocomial spread of the glycopeptide-intermediate S . aureus (GISA) strains has not been observed in US hospitals or in Europe, spread of GISA strains has apparently occurred in Japan . Laboratory studies have indicated that the disk diffusion test, the Stoke's method, and several automated methods of antimicrobial susceptibility testing do not detect GISA strains . The requirement to choose from a relatively small number of acceptable techniques for screening may well influence the ability of laboratories to conduct surveillance for these organisms . Finally, the isolation of such strains in three geographically distinct regions suggests that this phenomenon will continue to occur worldwide. Folia Med (Plovdiv), 1999, 41(3), 40 - 4 Mucocutaneous infections in hematological malignancies; Grudeva-Popova J et al.; INTRODUCTION: Mucocutaneous infections of diverse etiology are frequent complications in patients with hematological malignancies . OBJECTIVE: To study the incidence, predisposing factors, microbiology, and primary clinical manifestations of these complications in oncohematological patients . MATERIAL AND METHODS: The study included 862 patients (394 females and 468 males) with hematological malignancies, treated in the Clinic of Hematology of the Higher Medical Institute, Plovdiv from 1990 to 1998 . The patients were divided into three groups according to the primary disorder: lymphoproliferative disorders (541 patients), myeloproliferative disorders (296 patients), and bone marrow insufficiency (25 patients) . RESULTS AND DISCUSSION: The mucocutaneous infections studied included most often abscesses and pyodermias . The predominant isolate was Staphylococcus aureus . Viral infections were caused mainly by herpes simplex virus type I and, less frequently, varicella zoster virus with intercostal localization . Candida species was isolated predominantly in oropharyngeal and esophageal mycoses . CONCLUSION: Prolonged cytostatic-induced neutropenia and suppressed cellular immune response are the principal factors for the infectious complications in hematological malignancies . The early clinical diagnosis and prompt etiological treatment of the mucocutaneous infections are crucial for the prophylaxis of the systemic infectious complications. Prog Urol, 1999 Dec, 9(6), 1106 - 10 {IgA deficiency and multiple Staphylococcus aureus perirenal abscesses . A case report}; Gaschignard N et al.; The authors report the case of a patient with IgA deficiency who presented with a low-grade renal abscess . Due to the atypical nature of the clinical features and imaging, the diagnosis was not established prior to surgery . Despite radical surgery, the abscess recurred several weeks later . In the light of this case, the authors discuss the various possible diagnoses that must be considered in the presence of these types of images and the therapeutic approach during initial management . They discuss the various immune deficiencies to be investigated in a particular infectious context. J Pediatr, 2000 Feb, 136(2), 176 - 80 Defective interleukin-12/interferon-gamma pathway in patients with hyperimmunoglobulinemia E syndrome; Borges WG et al.; OBJECTIVE: Patients with the hyperimmunoglobulinemia E (hyper-IgE) syndrome are reported to have defective production of interferon gamma (IFN-gamma) . Because IFN-gamma is a major activator of polymorphonuclear leukocytes (PMNs), this could result in defective PMN chemotaxis and markedly elevated IgE levels because of the unopposed action of interleukin (IL)-4 . IL-12, an important enhancer of IFN-gamma production, also suppresses IgE production . This study assessed the IL-12/IFN-gamma pathway in patients with hyper-IgE syndrome . METHODS: Production of IL-12 and IFN-gamma by mononuclear cells from 10 patients with hyper-IgE syndrome in response to a number of stimuli was determined, as well as the effect of IL-12 on IFN-gamma release and cell proliferation . RESULTS: IL-12 and IFN-gamma production by the patients' cells was similar to that of control subjects independent of the stimulus used, except for Staphylococcus aureus, with which cells of patients with hyper-IgE syndrome released markedly less IFN-gamma (19.8%; P <.002) . The ability of recombinant IL-12 to enhance IFN-gamma release from patients' cells in response to all stimuli was, however, significantly lower than with control cells (12% to 51%; P <.03) . CONCLUSION: The lymphocytes of patients with hyper-IgE syndrome have an impaired response to IL-12, resulting in decreased IFN-gamma production, which may be of key importance in the pathogenesis of the immune abnormalities of hyper-IgE syndrome. J Immunol, 2000 Feb 15, 164(4), 1722 - 9 Inhibition of IL-12 production in human monocyte-derived macrophages by TNF; Ma X et al.; IL-12 is a pivotal cytokine that links the innate and adaptive immune responses . TNF-alpha also plays a key role in orchestrating inflammation and immunity . The reciprocal influence of these two inflammatory mediators on each other may have significant impact on the cytokine balance that shapes the type and extent of immune responses . To investigate the relationship between TNF-alpha and IL-12 production, we analyzed the effects of exposure of human monocyte-derived macrophages to TNF-alpha on LPS- or Staphylococcus aureus-induced IL-12 production in the presence or absence of IFN-gamma . TNF-alpha is a potent inhibitor of IL-12 p40 and p70 secretion from human macrophages induced by LPS or S . aureus . IL-10 is not responsible for the TNF-alpha-mediated inhibition of IL-12 . TNF-alpha selectively inhibits IL-12 p40 steady-state mRNA, but not those of IL-12 p35, IL-1alpha, IL-1beta, or IL-6 . Nuclear run-on analysis identified this specific inhibitory effect at the transcriptional level for IL-12 p40 without down-regulation of the IL-12 p35 gene . The major transcriptional factors identified to be involved in the regulation of IL-12 p40 gene expression by LPS and IFN-gamma, i.e., c-Rel, NF-kappaB p50 and p65, IFN regulatory factor-1, and ets-2, were not affected by TNF-alpha when examined by nuclear translocation and DNA binding . These data demonstrate a selective negative regulation on IL-12 by TNF-alpha, identifying a direct negative feedback mechanism for inflammation-induced suppression of IL-12 gene expression. Nat Biotechnol, 2000 Feb, 18(2), 163 - 7 Intracellular trehalose improves the survival of cryopreserved mammalian cells; Eroglu A et al.; We report that the introduction of low concentrations of intracellular trehalose can greatly improve the survival of mammalian cells during cryopreservation . Using a genetically engineered mutant of Staphylococcus aureus alpha-hemolysin to create pores in the cellular membrane, we were able to load trehalose into cells . Low concentrations (0.2 M) of trehalose permitted long-term post-thaw survival of more than 80% of 3T3 fibroblasts and 70% of human keratinocytes . These results indicate that simplified and widely applicable freezing protocols may be possible using sugars as intracellular cryoprotective additives. J Pathol, 2000 Feb, 190(2), 133 - 42 Trefoil factor family domain peptides in the human respiratory tract; dos Santos Silva E et al.; Trefoil factor family domain peptides (TFF) are thought to be involved in mucosal epithelial restitution and wound healing of the gastrointestinal tract and are up-regulated in ulceration and in a variety of solid tumours . It was hypothesized that TFFs are also expressed on mucosal surfaces of the human respiratory tract . Lung tissue, nasal polyps, and sputum samples from seven patients with cystic fibrosis (CF), two with chronic and acute bronchitis, and non-dysplastic material from two cases of bronchial adenocarcinoma were analysed for TFF expression by immunohistochemistry, immunofluorescence, western blot and RT-PCR . Expression of TFF1 and TFF3 was observed in material from all patients . TFFs were localized in goblet and ciliated cells, as well as in some submucosal cells of tracheobronchial tissues and nasal polyps from normal and CF individuals . In sputa of patients with CF and with chronic or acute bronchitis, TFF1 and TFF3 were detected by western blotting . Freshly cultivated nasal epithelial cells transcribed and secreted TFFs and mucins, whereas nasal cells cultivated for 6 weeks still expressed mucins, but not TFFs . Secreted TFFs and mucins also bound to the surface of Staphylococcus aureus in infected CF airways . In conclusion, TFF1 and TFF3 are expressed and secreted in normal and inflamed airways . The association of TFFs with bacteria may contribute to the anti-microbial mucociliary defence system . Cardiovasc Intervent Radiol, 2000 Jan-Feb, 23(1), 57 - 60 Subclavian arteritis and pseudoaneurysm formation secondary to stent infection; Malek AM et al.; Technically uncomplicated percutaneous angioplasty and stent placement of a left subclavian artery stenosis was performed in a 56-year-old man for treatment of subclavian steal syndrome and vertebrobasilar insufficiency . Six days later the patient was readmitted with Staphylococcus aureus bacteremia and stigmata of septic emboli isolated to the ipsilateral hand . Nine days later he had computed tomography (CT) evidence of a contrast-enhancing phlegmon surrounding the stent . Despite clinical improvement and resolution of bacteremia on intravenous antibiotic therapy, the phlegmon progressed, and at day 21 a pseudoaneurysm was angiographically confirmed . The patient underwent surgical removal of the stented arterial segment and successful autogenous arterial reconstruction . The possible contributory factors leading to stent infection were prolonged right femoral artery access and an infected left arm venous access . Although the role of prophylactic antibiotics remains to be defined, it may be important in cases where the vascular access sheath remains in place for a prolonged period of time. Mem Inst Oswaldo Cruz, 2000 Jan-Feb, 95(1), 29 - 33 Methicillin-resistant Staphylococcus aureus in human milk; Novak FR et al.; We collected and analyzed 500 samples of human milk, from five Brazilian cities (100 from each) to detect methicillin-resistant strains of Staphylococcus aureus (MRSA) producing enterotoxins . We found 57 strains of MRSA, and the mecA gene, responsible for resistance, was detected in all of them using a specific molecular probe . We examined 40 strains for the presence of four enterotoxins, after selecting a subset that included all strains from each region, except for the largest sample, from which 10 were randomly selected . Among these two presented enterotoxin B, and growth in human colostrum and trypicase soy broth . After 5 h of incubation at 37 degrees C, population sizes were already higher than 9.4 x 10(5) UFC/ml and enterotoxin was released into culture medium and colostrum . Our results stress the importance of hygiene, sanitary measures, and appropriate preservation conditions to avoid the proliferation of S . aureus in human milk. Clin Nucl Med, 2000 Feb, 25(2), 100 - 3 F-18 FDG uptake in breast infection and inflammation; Bakheet SM et al.; PURPOSE: Whole-body fluorine-18 fluorodeoxyglucose (F-18 FDG) positron emission tomography (PET) scanning has been useful in the management of breast cancer . However, F-18 FDG uptake sometimes has been associated with benign breast disease . Four cases are reported of F-18 FDG breast uptake caused by infectious or inflammatory mastitis that mimics malignant disease . METHODS AND RESULTS: Two women had F-18 FDG whole-body scans for the evaluation of a large breast mass after inconclusive results of ultrasonography . In both cases, intense focal F-18 FDG breast uptake was noted that mimicked breast cancer . Histologic examination showed, in one patient, chronic granulomatous infiltration that likely represented tuberculous mastitis, because she showed a good clinical response to empirical anti-tuberculous treatment . The second patient had lactational changes associated with acute inflammation, and the culture grew Staphylococcus aureus . The breast mass completely disappeared 3 weeks after a course of antibiotic treatment . The other two patients had staging F-18 FDG PET scans 1 and 12 months after lumpectomy for breast carcinoma to detect residual, recurrent, or metastatic disease . Both scans showed a ring-like uptake in the involved breast, with superimposed intense focal uptake suggesting tumor necrosis centrally and malignant foci peripherally . In both cases, histologic examination revealed hemorrhagic inflammation secondary to postsurgical hematomas and no evidence of malignancy . CONCLUSION: Acute or chronic infectious mastitis and postsurgical hemorrhagic inflammatory mastitis should be considered in patients who have a breast mass, especially those with a history of tenderness or surgery. Phytochemistry, 2000 Jan, 53(1), 93 - 6 An acylated phloroglucinol with antimicrobial properties from Helichrysum caespititium; Mathekga AD et al.; A new acylated form of a phloroglucinol with significant antimicrobial properties was isolated by bioactivity guided fractionation from Helichrysum caespititium (Asteraceae) . The structure elucidation, and conformation of the new phloroglucinol, 2-methyl-4-{2',4',6'-trihydroxy-3'-(2-methylpropanoyl) phenyl}but-2-enyl acetate, was established by high field NMR spectroscopic and MS data . The compound inhibited growth of Bacillus cereus, B . pumilus, B . subtilis and Micrococcus kristinae at the very low concentration of 0.5 microg/ml and Staphylococcus aureus at 5.0 microg/ml . Six fungi tested were similarly inhibited at low MICs, Aspergillus flavus and A . niger (1.0 microg/ml), Cladosporium chladosporioides (5 microg/ml), C . cucumerinum and C . sphaerospermum (0.5 microg/ml) and Phylophthora capsici at 1.0 microg/ml. J Clin Microbiol, 2000 Feb, 38(2), 866 - 9 Heterogeneous vancomycin resistance in methicillin-resistant Staphylococcus aureus strains isolated in a large Italian hospital; Marchese A et al.; Of 179 methicillin-resistant Staphylococcus aureus strains isolated from 1997 to 1998, two strains (1.1%) gave subclones for which the vancomycin MICs were 8 mg/liter . Pulsed-field gel electrophoresis showed identical restriction patterns for both isolates, suggesting transfer of a single clone between two different patients. J Clin Microbiol, 2000 Feb, 38(2), 846 - 50 Capsule expression by bovine isolates of Staphylococcus aureus from Argentina: genetic and epidemiologic analyses; Sordelli DO et al.; Staphylococcus aureus is an important cause of bovine mastitis worldwide, and effective preventive or therapeutic modalities are lacking . Although most human S . aureus isolates produce capsular polysaccharides (CPs), few reports have described the prevalence of capsules on bovine isolates . This information is important for the rational design of a vaccine for the prevention of staphylococcal mastitis . We serotyped 195 S . aureus strains isolated between 1989 and 1997 from the milk of mastitic cows in Argentina . Only 14 (7.1%) of the strains were serotype 5, and all were recovered between 1989 and 1992 . Thirteen serotype 8 strains were identified, and 12 of these were isolated between 1991 and 1994 . The remaining 168 isolates were nonreactive (NR) with CP serotype 5 (CP5)- or CP8-specific antibodies . Hybridization studies performed with genomic DNA from eight NR strains revealed that only three of them carried the capsule genes . Pulsed-field gel electrophoresis (PFGE) performed with 127 of the 195 S . aureus isolates revealed that most (86%) strains belonged to one of four major PFGE groups . Although 8 of 14 CP5 isolates showed a common PFGE pattern (arbitrarily defined as A1), 31 other A1 isolates from the same time period (1989 to 1992) were not CP5 positive . In contrast, only nine PFGE type B3 isolates were recovered between 1990 and 1994, and eight of these were positive for CP8 (P < 0.0003) . The results of this study underscore the variability in capsule expression by S . aureus strains isolated from different geographical regions and cast doubt on the roles of CP5 and CP8 in the pathogenesis and immunoprophylaxis of bovine mastitis in Argentina. Kidney Int, 2000 Feb, 57(2), 476 - 86 Accessory role of human peritoneal mesothelial cells in antigen presentation and T-cell growth; Hausmann MJ et al.; BACKGROUND: To assess the role of human peritoneal mesothelial cells (HPMCs) in the generation of an immune response during peritonitis, we tested their ability to activate T-cells by antigen presentation (AP) and by the secretion of interleukin-15 (IL-15) . IL-15 is a potent leukocyte activator that stimulates the proliferation of CD4+, CD8+, and B and natural killer (NK) cells . METHODS: HPMCs and mononuclear cells were derived from six volunteer patients who underwent elective abdominal surgery . Flow cytometry was used to analyze human lymphocyte antigen-DR (HLA-DR), intercellular adhesion molecule-1 (ICAM-1), and B7 molecules on HPMCs . Affinity-purified CD4 cells were used for AP assays . We used a specific enzyme-linked immunosorbent assay to detect interferon-gamma (IFN-gamma), IL-2, and IL-15 protein and reverse transcription-polymerase chain reaction for mRNA analysis . RESULTS: HPMCs expressed HLA-DR molecules following IFN-gamma treatment . ICAM-1 molecules were expressed at high levels, and B7-1 and B7-2 molecules could not be detected . The accessory function of HPMCs was assayed by T-cell stimulation using anti-CD3 antibodies (OKT3) . HPMCs were essential for a significant OKT3-induced T-cell proliferation . Anti-ICAM-1 antibodies blocked OKT3-induced proliferation . HPMCs served as effective antigen-presenting cells when Tetanus toxoid (TT) or Staphylococcus aureus-alpha-toxin were used as antigens . IFN-gamma, IL-2, and IL-15 accumulated during AP reactions . We found that IL-15 is produced by HPMCs, and IFN-gamma up-regulated its mRNA levels and protein secretion in a dose-dependent manner . We also detected IL-15 in the peritoneal effluent of patients undergoing continuous peritoneal dialysis treatment . In patients suffering from peritonitis, IL-15 levels were elevated (35.0 +/- 6.0 pg/mL, N = 10) as compared with noninfected patients (16.2 +/- 4.0 pg/mL, N = 7) . CONCLUSIONS: HPMCs participate in the peritoneal immune response against invading pathogens by AP . For this process, ICAM-1 is the major accessory molecule . In addition, HPMCs may contribute to T-cell activation by secretion of IL-15. J Invest Dermatol, 2000 Feb, 114(2), 281 - 8 Impaired responses of peripheral blood mononuclear cells to staphylococcal superantigen in patients with severe atopic dermatitis: a role of T cell apoptosis; Yoshino T et al.; Staphylococcus aureus colonization is an almost universal feature of atopic dermatitis . In order to investigate the role of staphylococcal enterotoxin B in the pathogenesis of atopic dermatitis, we assessed the correlation between clinical disease severity and proliferative response of peripheral blood mononuclear cells to staphylococcal enterotoxin B in patients with atopic dermatitis . Peripheral blood mononuclear cells from patients with mild atopic dermatitis showed significantly increased proliferative responses to staphylococcal enterotoxin B compared to controls . In contrast, peripheral blood mononuclear cells from patients with severe atopic dermatitis showed markedly suppressed proliferative responses . Additionally, longitudinal evaluation of peripheral blood mononuclear cell samples from the same patient demonstrated that proliferative responses were suppressed only at times of severe disease exacerbation . Mixing experiments, using autologous T cells and antigen presenting cells that were isolated at different time points from the same patient, demonstrated that T cells of severe atopic dermatitis patients were dysfunctional, but their antigen presenting cell function remained intact . We found no significant differences of interleukin-2 levels in the culture supernatants between healthy controls and atopic dermatitis groups . Fluorescence-activated cell sorter analysis for APO2.7 antigen, an early apoptosis cell marker, demonstrated that approximately 60% of staphylococcal-enterotoxin-B-stimulated T cells expressed APO2.7 antigen in severe atopic dermatitis cases . By contrast, 5%-20% of T cells expressed APO2.7 after staphylococcal enterotoxin B stimulation in cases of mild atopic dermatitis and in healthy controls . Nuclear staining with Hoechst 33258 also showed approximately 40% apoptotic cells in the CD19-CD16-PBMC of severe atopic dermatitis patients, compared with only 5%-10% in the mild atopic dermatitis group and in healthy controls . Blocking monoclonal antibody to Fas ligand partially prevented the staphylococcal-enterotoxin-B-induced apoptosis detected by APO2.7 expression and Hoechst 33258 staining . Suppressed proliferation of peripheral blood mononuclear cells in severe atopic dermatitis patients may be secondary to T cell death by apoptosis . These results suggest that an infection of S . aureus producing staphylococcal enterotoxin B may play a role in aggravation of atopic dermatitis by inducing apoptosis in T cells. Eur J Biochem, 2000 Feb, 267(3), 919 - 26 Molecular characterization of a human scavenger receptor, human MARCO; Elshourbagy NA et al.; Murine MARCO has been identified recently in subsets of macrophages located in the peritoneum, marginal zone of the spleen, and the medullary cord of lymph nodes, where it has been proposed that it serves as a bacteria-binding receptor . A scavenger receptor family member with an extended collagenous domain, murine MARCO has also been demonstrated in atherosclerotic lesions of susceptible mice . We report here the identification, tissue and chromosomal localization, and pharmacological characterization of human (h)MARCO . hMARCO was identified from a macrophage cDNA library by electronic screening with the murine MARCO sequence . Nucleotide sequence analysis confirmed that the full-length hMARCO clone encoded a 519-amino acid protein sharing 68.5% identity with murine MARCO . RNA blot analysis indicated that the hMARCO transcript is 2.0 kb in length and is predominantly expressed in human lung, liver, and lymph nodes . Radiation hybrid mapping localized hMARCO to chromosome 2q14 . Ligand-binding studies of COS cells expressing hMARCO demonstrated significant specific binding of both Escherichia coli and Staphylococcus aureus . In contrast, the hMARCO receptor expressed in COS cells did not specifically bind the scavenger receptor ligand acetylated low-density lipoprotein (LDL), despite its similarity to the elongated collagen-like binding domain of the macrophage scavenger receptor . In addition, acetylated (Ac)LDL and oxidized (Ox)LDL did not inhibit E . coli binding to hMARCO . These data suggest that hMARCO may play an important role in host defense, but it has no obvious role in the accumulation of modified lipoproteins during atherogenesis. Clin Endocrinol (Oxf), 2000 Jan, 52(1), 43 - 9 CSF rhinorrhoea following treatment with dopamine agonists for massive invasive prolactinomas; Leong KS et al.; OBJECTIVE: The management of CSF rhinorrhoea following dopamine agonist (DA) treatment for invasive prolactinomas is difficult and there is no clear consensus for its treatment . Our objective was therefore to investigate the different treatments for this condition . DESIGN AND PATIENTS: We examined the case notes of five patients with invasive prolactinomas and CSF rhinorrhoea following DA treatment . The different ways in which this complication had been managed is detailed along with a review of the literature . RESULTS: Five patients aged 24-67 years (3 male) with massive invasive prolactinomas (serum prolactin 95000-500000 mU/l) eroding the skull base were treated with dopamine agonists (3 bromocriptine, 1 cabergoline and 1 both) . CSF rhinorrhoea developed in all patients between 1 week and 4 months after commencing dopamine agonist treatment . In two patients (cases 1 and 4), CSF rhinorrhoea ceased within a few days of stopping bromocriptine but restarted when treatment was resumed . One of these (case 4), a 67-year-old woman had no further treatment and CSF leakage stopped completely . She died of unrelated medical problems 3 years later . In one patient staphylococcus aureus meningitis and pneumocephalus developed as a complication of CSF rhinorrhoea . Three patients had endoscopic nasal surgery to repair the fistula using muscle grafts, and to decompress the pituitary tumour, with success in two . One patient had intracranial surgery and dural repair, which was successful in sealing the leak . CONCLUSIONS: We suggest that surgery as soon as is feasible is the treatment of choice for the repair of a CSF leak following dopamine agonist treatment . An additional strategy is the withdrawal of dopamine agonist to allow tumour re-growth to stop the leak. J Dermatol Sci, 1999 Dec, 22(1), 62 - 5 Calcium oxide and magnesium oxide inhibit plasma coagulation by Staphylococcus aureus cells at the lower concentration than zinc oxide; Akiyama H et al.; We examined the effect of ceramic powder slurries on the coagulation of plasma by Staphylococcus aureus cells . Plasma coagulation by S . aureus strains or their cultured supernatant was inhibited in the plasma with 0.12% calcium oxide or 0.25% magnesium oxide after incubation for 24 h at 37 degrees C . Inhibition of plasma coagulation by calcium oxide and magnesium oxide was observed at the lower concentration than zinc oxide. J Bacteriol, 2000 Feb, 182(4), 1074 - 9 Cloning, characterization, and inactivation of the gene pbpC, encoding penicillin-binding protein 3 of Staphylococcus aureus; Pinho MG et al.; The gene pbpC from Staphylococcus aureus was sequenced: it encodes a 691-amino-acid protein with all of the conserved motifs of a class B high-molecular-weight penicillin-binding protein (PBP), including the transpeptidase conserved motifs SXXK, SXN, and KTG . Insertional inactivation of pbpC and introduction of the intact gene in a laboratory mutant missing PBP 3 showed that the pbpC gene encodes the staphylococcal PBP 3 . Inactivation of pbpC caused no detectable change in the muropeptide composition of cell wall peptidoglycan and had only minimum, if any, effect on growth rates, but caused a small but significant decrease in rates of autolysis . Cells of abnormal size and shape and disoriented septa were produced when bacteria with inactivated pbpC were grown in the presence of a sub-MIC of methicillin. Enferm Infecc Microbiol Clin, 1999 Dec, 17(10), 498 - 505 {The prevalence of methicillin-resistant Staphylococcus aureus phagotype 95 in the Hospitales Vall d'Hebron of Barcelona}; del Valle O et al.; BACKGROUND: In our hospital endemic methicillin resistant Staphylococcus aureus (MRSA) has been documented since 1971, with epidemic and interepidemic periods . During these years phage groups I, I-III, and non-typable were found by the international set of phages Phage group 95 (F95) was unusual between 1986 (when phage typing was first available) and 1991, with prevalence of 5.2% (mean), and 100% of sensibility to methicillin . In November 1991 appeared the first MRSA F95 strain, and its prevalence has been increasing until 1997 . MATERIAL AND METHODS: We have studied 133 strains of MRSA F95 isolated from 87 patients, 39 of them hospitalized in the General Hospital (HG), 38 in Traumatology Hospital (HT) and 8 in the Children's Hospital (HI) . Two of these patients had successive stancies in HG and HT . Antimicrobial susceptibility was determined by the disk diffusion method and microdilution to check oxacillin resistance . Moreover these method we have maked: detection of mecA, phage typing with the international set of phages and study of the PGFE patterns by digestion of chromosomic DNA with Smal . RESULTS: The percentage of methicillin resistance in F95 strains was increased since the appear of the first strain between 8.3% in 1991 to a maximum of 76.9% in 1995, we had a descens to 13.7% in 1996 but 1997 can back to augment it to 72.5% . MICs for oxacillin of these strains were low (< or = 64 mg/l to 87.4% of strains), and all of them were mecA positive, 78.1% of them were resistant to macrolides, 96.5% to tobramycin and 84.9% to quinolones, but only 10.5% to gentamicin, 4.7% were resistant to cotrimoxazol, 1.2% to fosfomycin and 2.5% to rifampin . All of them were sensible to doxycycline, and vancomycin . The pulse field gel electrophoresis showed 7 restriction patterns in MRSA F95, 73.8% of strains correspond to one of them (B), spreading from the spinal cord injury unit and prevalent in HT; and 10.8% to another (C), the first that appear, spreading from the neurosurgical unit and with high prevalence in HG . 6.9% has pattern J a B subtype that appear in broth HG and HT . Pattern E is prevalent in HI it was spread from neonatology unit . CONCLUSIONS: The emergence in a Center with endemic resistance of new strains of MRSA, not all of them of the same clone, with characteristic resistance pattern to antibiotics and in convivence with other phage groups is one demonstration of genetic variability of SAMR in our entorn. Adv Perit Dial, 1998, 14, 173 - 9 Nitric oxide is a marker of peritonitis in patients on continuous ambulatory peritoneal dialysis; Choi KC et al.; The aim of this study was to determine whether nitric oxide (NO) production is altered during peritonitis in patients receiving continuous ambulatory peritoneal dialysis (CAPD), and if so, whether there is an association between this alteration and the severity and prognosis of CAPD-induced peritonitis . The study population comprised 30 patients with 30 episodes of peritonitis . Thirteen patients without peritonitis were used as CAPD-control, and eighteen patients with normal renal function were used as normal-control . Total NO metabolites (NOx; nitrite + nitrate) were measured by the Griess method to reflect nitric oxide production . Peritoneal dialysate effluent and plasma were collected from 30 patients during episodes of peritonitis every day for the first 3 days, and then every 3 days for 2 weeks or until the patients were discharged . Plasma NOx levels in the control, CAPD-control, and CAPD-peritonitis groups were 87.0 +/- 11.5, 163.0 +/- 30.7 and 146.3 +/- 18.1 microM, respectively . Dialysate NOx levels in the CAPD-control and CAPD-peritonitis groups were 91.8 +/- 13.1 and 103.8 +/- 14.1 microM, respectively, and dialysate NOx levels did not differ between the two groups . The peak dialysate/plasma (D/P) ratios during the acute phase exceeded 1.0 in 46.7% of the patients of the CAPD-peritonitis group . The D/P ratios of NOx levels before and after treatment were 1.03 +/- 0.07 and 0.56 +/- 0.05, respectively . On the contrary, NOx levels in dialysate after treatment were not decreased, but those in plasma were increased after effective treatment . The peak D/P ratio increased 2.1-fold in the bacterial peritonitis group and 2.3-fold in the fungal peritonitis group, compared with the CAPD-control group . The lowest D/P ratios after treatment were similar to those in the CAPD-control group in patients with effective treatment, but remained 1.5-fold higher in patients for whom treatment was ineffective . In the evolutional study, the D/P ratios of NOx levels gradually declined to CAPD-control group levels (6.6 +/- 2.5 days) after effective antibiotic treatment, but it took longer for leukocyte counts in the peritoneal dialysate effluents (3.8 +/- 1.2 days) to normalize . In 5 patients with refractory peritonitis (Candida infection in three, Staphylococcus aureus infection in two), the D/P ratios of NOx levels remained elevated by 1.5-fold despite treatment, and the catheters were removed . These results suggest that dialysate NOx may be influenced not only by local NO production, but also by plasma NO or NOx diffusion . Therefore, we can suppose that the D/P ratio of NOx levels provides more clinical significance than dialysate NOx levels only . In conclusion, the D/P ratios of NOx levels may serve as a marker to assess the severity of peritoneal inflammation, treatment efficacy, and progression of refractory peritonitis in CAPD patients with peritonitis. Adv Perit Dial, 1998, 14, 137 - 41 Continuous cycler therapy, manual peritoneal dialysis therapy, and peritonitis; Troidle LK et al.; An increasing number of patients are prescribed a continuous-cycling regimen because standard manual peritoneal-dialysis exchanges alone are not sufficient in achieving adequate dialysis as defined by the Dialysis Outcome Quality Initiative . Consequently, the number of patients on continuous-cycler therapy is increasing . There is controversy as to whether there are differences in the development of peritonitis between patients maintained on manual therapy and those on continuous cycling therapy . As a result, we retrospectively reviewed the charts of all cycler peritoneal dialysis (CPD) patients maintained on either manual peritoneal dialysis (Baxter UltraBag; Group I) or continuous cycler peritoneal dialysis (Baxter HomeChoice Cycler; Group II) between 1 June 1994 and 31 December 1996 . A total of 239 patients were in Group I and 106 in Group II . Both groups were similar in age, race, gender, and presence of diabetes mellitus, coronary artery disease, peripheral vascular disease, and gastrointestinal disease . There was no difference in the overall rate of peritonitis between the two groups of patients {1 episode in 10.4 patient-months (Group I) vs . 1 in 10.0 patient-months (Group II); -0.01843 to 0.02619} . The rates of Staphylococcus aureus peritonitis {1 episode in 48.5 patient-months (Group I) vs . 1 in 141.8 patient months (Group II); -0.06152 to -1.1689}; polymicrobial peritonitis {1 episode in 278.8 patient-months (Group I) vs . 1 in 1134 patient months (Group II): -0.0079 to -0.0478}, and fungal peritonitis (1 episode in 202.7 patient-months (Group I) vs . no episodes (Group II); 0.00202 to 0.00785} were significantly lower among patients maintained on the Baxter HomeChoice Cycler . The rate of gram-negative peritonitis was higher among patients maintained on the Baxter HomeChoice Cycler, but this difference was not statistically significant {1 episode in 82.6 patient-months (Group I) vs . 1 episode in 45.4 patient months (Group II); 0.4723 to -0.0248} . We conclude that individual rates of peritonitis were different for patients maintained on either manual or continuous CPD therapy, while the overall rate of peritonitis was found to be similar for both groups of patients . The finding that there may be a difference with the gram-negative peritonitis rate is important since gram-negative peritonitis has been shown to have a more severe outcome in terms of morbidity, mortality, and patient dropout from CPD therapy . A larger, randomized, multicenter study comparing the rates of gram-positive, gram-negative, fungal, and polymicrobial peritonitis is warranted. Surg Today, 2000, 30(1), 16 - 21 Preoperative intranasal mupirocin ointment significantly reduces postoperative infection with Staphylococcus aureus in patients undergoing upper gastrointestinal surgery; Yano M et al.; Nasal carriage of Staphylococcus aureus including methicillin-resistant S . aureus (MRSA) is associated with an increased risk for postoperative staphylococcal infection . This study was conducted to investigate the effect of preoperative nasal mupirocin treatment on the postoperative infections in patients undergoing upper gastrointestinal surgery . The intervention group consisted of 141 consecutive patients who underwent upper gastrointestinal surgery between March 1, 1997, and February 28, 1998 . The patients in the intervention group were treated with intranasal mupirocin three times a day for 3 consecutive days before surgery . The incidence of postoperative staphylococcal infections in the intervention group was then compared with that of the historical control group . The control group consisted of 128 consecutive patients who underwent upper gastrointestinal surgery without mupirocin treatment between January 1, 1996 and December 31, 1996 . The postoperative staphylococcal infection rate in the control group (11.7%) was significantly higher (P < 0.001) than in the intervention group (0.71%) . The postoperative MRSA infection rate was significantly reduced by the intervention (control group 7.0% and intervention group 0%; P < 0.01) . These results suggest that preoperative nasal eradication of S . aureus with mupirocin thus appears to be an effective measure to prevent postoperative staphylococcal infection in patients undergoing upper gastrointestinal surgery. Ter Arkh, 1999, 71(12), 28 - 31 {Role of staphylococcus aureus hemolytic toxin-alpha in pathogenesis of infectious endocarditis: studies in vitro}; Baliakina EV et al.; AIM: To study effects of alpha-toxin (AT) Staphylococcus aureus on human platelets and endothelial cells . MATERIALS AND METHODS: Concentrations of intracellular calcium in human platelets and endothelial cells were estimated by fluorescence, phosphoinositide metabolism in the endothelial cells was studied using 3H-myoinositole . RESULTS: AT induced a dose-dependent increase of intracellular calcium in blood and vascular cells, stimulates dose-dependent formation of inositol phosphates in endothelial cells . CONCLUSION: AT action on the platelets and endothelial cells results in a significant receptor-independent rise in concentration of intracellular calcium, activation of phosphoinositide metabolism, death of cells . These data support the hypothesis that the platelet and endothelial cell damage is mostly due to the passive Ca2+ influxvia pores formed by AT in cellular membrane. J Leukoc Biol, 2000 Jan, 67(1), 40 - 5 Effect of age on human neutrophil function; Wenisch C et al.; Neutrophil phagocytosis, reactive oxygen intermediate production (intra- and extracellular), neutrophil bactericidal activity, and chemotaxis/chemokinesis were assessed in three age groups: 21-36, 38-56, and 62-83 years . A significant age-dependent reduction in the number of phagocytized Escherichia coli per neutrophil (measured by acridine orange staining) and Staphylococcus aureus phagocytosis (measured by flow cytometry) was seen (r = 0.669 and r = 0.684, P<0.001 for both) . These findings correlated with an age-dependent increase in intracellular calcium concentrations in resting neutrophils (r = 0.698, P<0.001) and a reduced hexose uptake (r = 0.591, P<0.01) . In addition, a significant reduction in the intracellular reactive oxygen production was seen after stimulation with S . aureus (P<0.001) with increasing age . In contrast, no differences between the groups in reactive oxygen production was seen after stimulation with E . coli . The neutrophil bactericidal activity was impaired with increasing age (64+/-4% of the phagocytized bacteria were killed in group 1; 66+/-2 in group 2, and 59+/-6 in group 3; P<0.01) . In addition, a trend toward a reduced neutrophil chemotaxis was seen with increasing age (P = 0.022) . The findings suggest that increased intracellular calcium concentrations in resting neutrophils and/or a reduced hexose uptake result in reduced phagocytic ability and decreased bactericidal activity of neutrophils in the elderly. Eur J Cardiothorac Surg, 1999 Dec, 16(6), 613 - 8 Does focal destruction of the thoracic aorta wall by Staphylococcus aureus lead to the development of infected aneurysms? An experimental study; Fantidis P et al.; OBJECTIVE: The infrequency of infected aneurysms suggests that either infection of segments of the aortic wall is uncommon, or that infections do not always lead top infected aneurysm formation . The purpose of the study was to determine whether focal Staphylococcus aureus infection of aortic wall segments leads consistently to the development of infected aneurysms and to evaluate the segments in which infection did not lead to the infected aneurysm formation . METHODS: Twenty pigs were inoculated with 0.1 ml of a Staphylococcus aureus inoculum in three segments of the thoracic aorta wall (study group) . In another 10 pigs, 0.1 ml of saline solution was injected in three segments of the thoracic aorta wall (control group) . Study group: histological abnormalities and bacterial culture of the inoculation sites were evaluated at 10 days (n = 5 pigs), 30 days (n = 5 pigs), and 90 days (n = 10 pigs) . Control group: histological abnormalities were evaluated at 10 days (n = 5 pigs) and 90 days (n = 5 pigs) . RESULTS: Study group: infected aneurysms developed in only two animals killed at 30 days . At 90 days, destruction of the elastic tissue, scar tissue and neointima formation were found in all the aortic segments studied . Control group: no significant changes were found in any of the segments evaluated . CONCLUSION: In our experimental model, acute local infection by S . aureus caused the development of infected aortic aneurysm in only 10% of the animals . In the remaining 90%, healing of the site of infection followed resolution of the infection. Microb Drug Resist, 1999 Winter, 5(4), 253 - 7 Inactivation of the methicillin resistance gene mecA in vancomycin-resistant Staphylococcus aureus; Sieradzki K et al.; Acquisition of high-level resistance to vancomycin in the laboratory mutant VM50 (vancomycin MIC increased from 1.5 to 100 microg/ml) was accompanied by the appearance of a heterogeneous phenotype and a virtual loss in methicillin resistance: in most cells of cultures of VM50 the methicillin MIC of the parental strain was reduced from 800 to 1.5 microg/ml with only a subpopulation (10(-5)) retaining methicillin resistance at near the parental level (MIC of 400 microg/ml) . Interestingly, the vancomycin MIC of this subpopulation was less (25 microg/ml) than that of VM50 (100 microg/ml) . A similar antagonism between methicillin and vancomycin resistance levels was observed upon introduction of an intact mecA into VM50 on a plasmid vector: methicillin resistance of the majority of cells increased from 1.5 to 100 microg/ml while the vancomycin MIC declined from 100 to 12/25 microg/ml . Membrane preparations from mutant VM50 showed no detectable penicillin-binding protein (PBP) 2A by the fluorographic assay . Sequencing of the mecA gene resident in mutant VM50 indicated the presence of a 19-bp duplication between nucleotide residues 280-298, leading to the generation of a stop codon TAA starting at nucleotide position 286. J Cataract Refract Surg, 2000 Jan, 26(1), 140 - 1 Infectious keratitis after photorefractive keratectomy in a comanaged setting; Heidemann DG et al.; A 48-year-old man had simultaneous bilateral photorefractive keratectomy (PRK) . The surgeon who performed the PRK did not see the patient in follow-up, and there was confusion regarding the comanaging doctor . Therefore, the patient was not examined immediately postoperatively . Several days later, he was hospitalized for an unrelated, painful orthopedic problem and heavily sedated . Seven days after the PRK, an ophthalmologist was consulted for ocular irritation and discharge . Examination showed bilateral, purulent conjunctivitis and severe infectious keratitis in the left eye . The patient was treated with periocular and topical antibiotics . Corneal cultures yielded Staphylococcus aureus . The keratitis resolved slowly, leaving the patient with hand motion visual acuity . A corneal transplant and cataract extraction was performed 15 months later, resulting in a best corrected visual acuity of 20/400 because of glaucomatous optic nerve damage . Severe infectious keratitis may occur after PRK . Poor communication between the surgeon, comanaging doctor, and patient may result in treatment delay. Biochem J, 2000 Feb 1, 345 Pt 3, 611 - 9 A bone sialoprotein-binding protein from Staphylococcus aureus: a member of the staphylococcal Sdr family; Tung H et al.; Staphylococcus aureus bacteria, isolated from bone and joint infections, specifically interact with bone sialoprotein (BSP), a glycoprotein of bone and dentine extracellular matrix, via a cell-surface protein of M(r) 97000 {Yacoub, Lindahl, Rubin, Wendel, Heinegard and Ryden, (1994) Eur . J . Biochem . 222, 919-925} . Amino acid sequences of seven trypsin fragments from the 97000-M(r) BSP-binding protein were determined . A gene encoding a protein encompassing all seven peptide sequences was identified from chromosomal DNA isolated from S . aureus strain O24 . This gene encodes a protein with 1171 amino acids, called BSP-binding protein (Bbp), which displays similarity to recently described proteins of the Sdr family from S . aureus . SdrC, SdrD and SdrE encode putative cell-surface proteins with no described ligand specificity . Bbp also shows similarity to a fibrinogen-binding protein from S . epidermidis called Fbe . A serine-aspartic acid repeat sequence was found close to the cell-wall-anchoring Leu-Pro-Xaa-Thr-Gly sequence in the C-terminal end of the protein . Escherichia coli cells were transformed with an expression vector containing a major part of the bbp gene fused to the gene for glutathione S-transferase . The affinity-purified fusion protein bound radiolabelled native BSP, and inhibited the binding of radiolabelled BSP to staphylococcal cells . Serum from patients suffering from bone and joint infection contained antibodies that reacted with the fusion protein of the BSP-binding protein, indicating that the protein is expressed during an infection and is immunogenic . The S . aureus Bbp protein may be important in the localization of bacteria to bone tissue, and thus might be of relevance in the pathogenicity of osteomyelitis. J Hand Surg {Am}, 2000 Jan, 25(1), 173 - 5 Methicillin-resistant Staphylococcus aureus in a finger felon; Connolly B et al.; Methicillin-resistant Staphylococcus aureus is an increasingly prevalent nosocomial pathogen that presents therapeutic challenges . We report an incidence of methicillin-resistant S aureus in a felon . The biochemical and clinical characteristics of methicillin-resistant S aureus are reviewed . The alarming increase of this organism in various types of infections demands the attention of all surgeons and emphasizes the importance of early surgical drainage and culture of pus in all cases of infection . (J Hand Surg 2000; 25A:173-175 . FEMS Immunol Med Microbiol, 2000 Feb, 27(2), 95 - 8 Frequency of nasal and wound isolates of Staphylococcus aureus associated with TSST-1 production in Jordanian population; Daghistani HI et al.; A total of 110 Staphylococcus aureus isolates were obtained from nasal carriers and wound infections of Jordanian population . The isolates were identified by cultural and biochemical methods . The nasal carrier rate of S . aureus among individuals was 22.7% . In comparison with the nasal S . aureus isolates the wound isolates did not produce significantly more virulence factors except DNase . Toxic shock syndrome toxin-1 production was higher among the S . aureus nasal isolates (40%) as compared with the wound isolates (26%) detected by an ELISA method which proved to be uniformly more sensitive than the immunodiffusion optimal sensitivity plate (OSP) method. Arch Biochem Biophys, 2000 Feb 1, 374(1), 3 - 7 Secretion and purification of recombinant beta1-4 galactosyltransferase from insect cells using pFmel-protA, a novel transposition-based baculovirus transfer vector; Zhou D et al.; The palette of transfer vectors available for generation of recombinant baculoviruses based on transposition-mediated recombination has been enlarged by constructing the pFmel-protA vector . The pFmel-protA plasmid includes the honeybee melittin secretion signal and a Staphylococcus aureus protein A fusion protein tag, which allows the secretion and purification of recombinant proteins . Using this system, the human beta1-4 galactosyltransferase-I protein was expressed in Sf9 insect cells at a level ranging from 22 to 28 U (4.8 to 6.0 mg)/L . The protein A tag enabled a simple monitoring of recombinant protein expression by enzyme-linked immunosorbent assay and Western blotting . Single step purification was achieved by immunoglobulin G affinity chromatography achieving a recovery yield of 28% and a specific activity of 1.9 U per mg of recombinant protein . Antimicrob Agents Chemother, 2000 Feb, 44(2), 294 - 303 Characterization of passage-selected vancomycin-resistant Staphylococcus aureus strains of diverse parental backgrounds; Pfeltz RF et al.; A series of 12 Staphylococcus aureus strains of various genetic backgrounds, methicillin resistance levels, and autolytic activities were subjected to selection for the glycopeptide-intermediate S . aureus (GISA) susceptibility phenotype on increasing concentrations of vancomycin . Six strains acquired the phenotype rapidly, two did so slowly, and four failed to do so . The vancomycin MICs for the GISA strains ranged from 4 to 16 microg/ml, were stable to 20 nonselective passages, and expressed resistance homogeneously . Neither ease of acquisition of the GISA phenotype nor the MIC attained correlated with methicillin resistance hetero- versus homogeneity or autolytic deficiency or sufficiency . Oxacillin MICs were generally unchanged between parent and GISA strains, although the mec members of both isogenic methicillin-susceptible and methicillin-resistant pairs acquired the GISA phenotype more rapidly and to higher MICs than did their susceptible counterparts . Transmission electron microscopy revealed that the GISA strains appeared normal in the absence of vancomycin but had thickened and diffuse cell walls when grown with vancomycin at one-half the MIC . Common features among GISAs were reduced doubling times, decreased lysostaphin susceptibilities, and reduced whole-cell and zymographic autolytic activities in the absence of vancomycin . This, with surface hydrophobicity differences, indicated that even in the absence of vancomycin the GISA cell walls differed from those of the parents . Autolytic activities were further reduced by the inclusion of vancomycin in whole-cell and zymographic studies . The six least vancomycin-susceptible GISA strains exhibited an increased capacity to remove vancomycin from the medium versus their parent lines . This study suggests that while some elements of the GISA phenotype are strain specific, many are common to the phenotype although their expression is influenced by genetic background . GISA strains with similar glycopeptide MICs may express individual components of the phenotype to different extents. Antimicrob Agents Chemother, 2000 Feb, 44(2), 272 - 7 Reversion of the glycopeptide resistance phenotype in Staphylococcus aureus clinical isolates; Boyle-Vavra S et al.; The recent identification of glycopeptide intermediate-resistant Staphylococcus aureus (GISA) clinical isolates has provided an opportunity to assess the stability of the glycopeptide resistance phenotype by nonselective serial passage and to evaluate reversion-associated cell surface changes . Three GISA isolates from the United States (MIC of vancomycin = 8 microg/ml) and two from Japan (MICs of vancomycin = 8 and 2 microg/ml) were passaged daily on nutrient agar with or without vancomycin supplementation . After 15 days of passage on nonselective medium, vancomycin- and teicoplanin-susceptible revertants were obtained from each GISA isolate as determined by broth dilution MIC . Revertant isolates were compared with parent isolates for changes in vancomycin heteroresistance, capsule production, hemolysis phenotype, coagulase activity, and lysostaphin susceptibility . Several revertants lost the subpopulations with intermediate vancomycin resistance, whereas two revertants maintained them . Furthermore, although all of the parent GISA isolates produced capsule type 5 (CP5), all but one revertant tested no longer produced CP5 . In contrast, passage on medium containing vancomycin yielded isolates that were still intermediately resistant to vancomycin, had no decrease in the MIC of teicoplanin, and produced detectable CP5 . No consistent changes in the revertants in hemolysis phenotype, lysostaphin susceptibility, or coagulase activities were discerned . These data indicate that the vancomycin resistance phenotype is unstable in clinical GISA isolates . Reversion of the vancomycin resistance phenotype might explain the difficulty in isolating vancomycin-resistant clinical isolates from the blood of patients who fail vancomycin therapy and, possibly, may account for some of the difficulties in identifying GISA isolates in the clinical laboratory. Infect Immun, 2000 Feb, 68(2), 973 - 6 Two allelic forms of the aureolysin gene (aur) within Staphylococcus aureus; Sabat A et al.; Proteinases of Staphylococcus aureus are emerging as potential virulence factors which may be involved in the pathogenecity of staphylococcal diseases . We describe here the structure of the gene encoding the metalloproteinase referred to as aureolysin . This gene occurs in two allelic forms and is strongly conserved among S . aureus strains, implying the possibility that the proteinase may have important housekeeping functions. Proc Natl Acad Sci U S A, 2000 Jan 18, 97(2), 617 - 22 Direct localization of a beta-subunit domain on the three-dimensional structure of Escherichia coli RNA polymerase; Opalka N et al.; To identify the location of a domain of the beta-subunit of Escherichia coli RNA polymerase (RNAP) on the three-dimensional structure, we developed a method to tag a nonessential surface of the multisubunit enzyme with a protein density easily detectable by electron microscopy and image processing . Four repeats of the IgG-binding domain of Staphylococcus aureus protein A were inserted at position 998 of the E . coli RNAP beta-subunit . The mutant RNAP supported E . coli growth and showed no apparent functional defects in vitro . The structure of the mutant RNAP was determined by cryoelectron microscopy and image processing of frozen-hydrated helical crystals . Comparison of the mutant RNAP structure with the previously determined wild-type RNAP structure by Fourier difference analysis at 20-A resolution directly revealed the location of the inserted protein domain, thereby locating the region around position 998 of the beta-subunit within the RNAP three-dimensional structure and refining a model for the subunit locations within the enzyme. J Lab Clin Med, 2000 Jan, 135(1), 43 - 51 Staphylococcus aureus adherence to thrombin-treated endothelial cells is mediated by fibrinogen but not by platelets; Shenkman B et al.; Recent studies emphasize the role of blood constituents in Staphylococcus aureus (S . aureus) adherence to subendothelial extracellular matrix . In the present study, the adherence of two strains of S . aureus (ATCC 29213 and RN 6390) grown to a postexponential phase to cultured human umbilical vein endothelial cells (EC-304) was examined . Under flow conditions (600 s(-1)), pretreatment of endothelial cells (ECs) with human alpha-thrombin (2 U/mL) significantly (2- to 4-fold) increased bacterial adherence to ECs . Adherence of both S . aureus strains to thrombin-treated ECs was similarly higher in the presence of whole blood, platelet-rich plasma, or platelet-poor plasma when compared with Tris-buffered saline solution (TBS) . Platelet inactivation in whole blood by prostaglandin E1 did not reduce the adherence rate . When ATCC 29213 bacteria were suspended in TBS containing increasing concentrations of fibrinogen at near-physiologic ranges (0.25 to 2 mg/mL), a dose-dependent increase in S . aureus adherence to thrombin-activated ECs was observed that reached a maximum level of about 12-fold . Fibronectin used at the above physiologic concentrations (12.5 to 100 microg/mL) enhanced bacterial adherence up to 2-fold . Von Willebrand factor (1 IU/mL) did not support bacterial adherence to ECs, either alone or in combination with fibrinogen . Inhibition of fibrin formation either by the Gly-Pro-Arg-Pro peptide or by hirudin increased bacterial adherence by 50% and 90%, respectively . Blockage of either ICAM-I, alpha5beta1, or alphavbeta3 receptors on ECs by appropriate monoclonal antibodies resulted in substantial inhibition of bacterial adherence (by 42%, 65%, and 72%, respectively) . Preincubation of S . aureus with a fibrinogen gamma-chain binding domain peptide led to 65% inhibition of adherence to ECs in the presence of fibrinogen . In contrast, preincubation of bacteria with the Arg-Gly-Asp-Ser peptide failed to affect their adherence . The data suggest that S . aureus adherence to the EC surface was (1) significantly enhanced by thrombin treatment of ECs, (2) not mediated by platelets, and (3) mediated by plasma fibrinogen, which binds to the bacteria via the C-terminus gamma-chain binding domain but not via the Arg-Gly-Asp sequence. J Lab Clin Med, 2000 Jan, 135(1), 32 - 42 Identification of plasma proteins adsorbed on hemodialysis tubing that promote Staphylococcus aureus adhesion; Francois P et al.; Risk factors for Staphylococcus aureus infections in patients undergoing hemodialysis include underlying disease, material-induced host defects, and the presence of vascular access catheters . To determine the specific contribution of various potentially adsorbed plasma components in promoting S aureus adhesion to shunt tubing during chronic hemodialysis, we quantified their respective amounts by Western immunoblotting and densitometry and estimated their individual adhesion-promoting activities with specific adhesion-modified bacterial mutants . Fibrinogen, which was the only component consistently present in tubing protein extracts from all patients, was adsorbed in significantly higher amounts on predialyzer than on postdialyzer tubing segments . In contrast, fibronectin and von Willebrand factor were irregularly present in patients' tubing, whereas vitronectin or thrombospondin remained undetectable in all samples . The contribution of fibrinogen in promoting S aureus attachment to hemodialysis tubing was demonstrated by (1) the significantly lower adhesion of a cIfA mutant of strain Newman compared with its parent; (2) the increased attachment of strain 8325-4 after complementation with the cloned cIfA gene on the multicopy plasmid pCF4; and (3) the general tendency for strains Newman and 8325-4(pCF4) to express higher attachment on predialyzer compared with postdialyzer tubing segments in relationship with the higher content of fibrinogen on the former material . However, the specific S aureus attachment-promoting activity of both prefilter and postfilter tubing-adsorbed fibrinogen were much lower than that of the native in vitro-adsorbed protein and may reflect masking or inactivation of the in vivo-adsorbed protein by unknown mechanisms. J Microbiol Immunol Infect, 1999 Sep, 32(3), 194 - 8 Susceptibility testing and clinical effect of fusidic acid in oxacillin-resistant Staphylococcus aureus infections; Liu CP et al.; One hundred and six oxacillin-resistant Staphylococcus aureus (ORSA) isolates collected from various clinical specimens at Mackay Memorial Hospital during the period from 1997 to 1998 were tested . The MICs of fusidic acid against ORSA isolates were in the range of 0.06 microgram/mL to 4 micrograms/mL . There was only one ORSA isolate (0.9%) resistant to fusidic acid in this study . Twenty-four patients with ORSA infection were enrolled into the study between July 1997 to June 1998 . Four patients without the evidence of sepsis received oral fusidic acid only . The other 20 patients with sepsis received intravenous glycopeptide first, and then the oral fusidic acid for 7 to 10 days . Oral fusidic acid was used for mild ORSA infection and achieved satisfactory clinical outcome . In severe ORSA infection, it was found that sequential therapy with fusidic acid had relapsed in three patients . We concluded that oral fusidic acid achieved a satisfactory outcome in our patients with mild ORSA infection . In such cases, fusidic acid may provide an effective alternative treatment of choice . The rare side effect of granulocytopenia and jaundice was noted in our case series . The side effect may be not so rare as previously thought, if it had been carefully looked for. J Biol Chem, 2000 Jan 21, 275(3), 1630 - 4 Isolation and characterization of proteins that bind to galactose, lipopolysaccharide of Escherichia coli, and protein A of Staphylococcus aureus from the hemolymph of Tachypleus tridentatus; Chiou ST et al.; In this study, we report the isolation and characterization of three novel hemolymph proteins that are believed to be involved in the innate immune response of horseshoe crabs, Tachypleus tridentatus . They include two closely related proteins, one that binds to the protein A of Staphylococcus aureus (PAP) and another that binds to the lipopolysaccharide of Escherichia coli (LBP) . PAP binds specifically to staphylococcal protein A (SpA) with a K(D) of 3.86 x 10(-5) M, whereas LBP binds to lipopolysaccharide (LPS) with a K(D) of 1.03 x 10(-6) M . Both PAP and LBP are glycoproteins with an apparent molecular mass of about 40 kDa . N-terminal sequences of PAP and LBP showed 61.9 and 72.2% identity, respectively, to tachylectin-3, a lectin isolated from the amebocyte of T . tridentatus, previously characterized by its affinity to the O-antigen of LPS and blood group A antigen (Muta, T., and Iwanaga, S . (1996) Curr . Opin . Immunol . 8, 41-47) . The third protein, a galactose-binding protein (GBP), was found to bind tightly to Sepharose CL-4B and could only be eluted from the column matrix with chaotropic agents, such as 4 M urea or 2 M guanidine hydrochloride . Further analysis indicated that GBP binds to D(+)-galactose with a K(D) of 2.47 x 10(-7) M . N-terminal sequence analysis showed that GBP shared a 50% identity with lectin L-6, identified in the granules of amebocyte of T . tridentatus . (Gokudan, S., Muta, T., Tsuda, R., Koori, K., Kawahara, T., Seki, N., Mizunoe, Y., Wai, S . N . , Iwanaga, S., and Kawabata, S . (1999) Proc . Natl . Acad . Sci . U . S . A . 96, 10086-10091) . Lectin-L6 and tachylectin-3 are nonglycosylated intracellular proteins with about half the molecular mass of PAP, LBP, and GBP . GBP also binds to PAP and LBP with K(D) values of 1.25 x 10(-7) and 1.43 x 10(-8) M, respectively, and this binding is enhanced about 10-fold upon the addition of SpA and LPS to form the GBP.PAP.SpA and GBP.LBP.LPS complexes, respectively. Antibiot Khimioter, 1999, 44(10), 16 - 9 {The microbiological aspects of infectious complications in the oncology clinic}; Dmitrieva NV et al.; At present 10 to 30 per cent of the microbial strains from cancer patients are problem ones: oxacillin resistant strains of Staphylococcus aureus, coagulase negative strains of Staphylococcus spp., aminoglycoside resistant strains of Escherichia coli, 3rd generation cephalosporin resistant strains of Klebsiella spp . and fungi of Candida which requires development of more rational approaches to antibacterial chemotherapy and prophylaxis of infectious complications . The infectious processes in the cancer patients proved to be highly polyetiological . Therefore, the study is significant for epidemiologic and therapeutic measures . Such an analysis in oncological clinic should be regular. Ann Surg, 2000 Jan, 231(1), 126 - 31 Technical refinement in adult-to-adult living donor liver transplantation using right lobe graft; Fan ST et al.; OBJECTIVE: To report the authors' experience with living donor liver transplantation in adults using right lobe liver grafts, performed by a modified technique . SUMMARY BACKGROUND DATA: The initial results of seven living donor liver transplants in adults using extended right lobe grafts were satisfactory, but serious complications occurred in two donors, and six recipients required repeat laparotomy . Another 11 similar operations were performed . Further evaluation was made with the aim of improving the postoperative outcome . METHODS: From December 1996 to August 1998, 11 patients underwent living donor liver transplantation using right lobe grafts . The first four patients underwent surgery using methods previously designed and the next seven underwent a modification designed to minimize devitalized tissues on the liver transection surface, improve hepatic venous drainage, and reduce the number of hepatic duct orifices . RESULTS: There were no donor deaths . Donor complications included cholestasis (n = 1) and minor wound infection (n = 1) . All the first four recipients required a repeat laparotomy for infected necrotic liver transection surface (n = 1), acute pancreatitis (n = 1), hepatic vein thrombosis (n = 1), and leakage from one of the two bilioenteric anastomoses (n = 1) . The patient with hepatic vein thrombosis died . In the last seven recipients, all of whom survived the operation, one required a repeat laparotomy with the discovery of a methicillin-resistant Staphylococcus aureus culture of fibrinous exudate at the left subphrenic peritoneum, and another had right hepatic duct stump necrosis . The latter was likely related to hypovolemic shock secondary to bleeding from the right saphenous vein on removal of a hemofiltration catheter . Comparison of the incidence of repeat laparotomy between the first four and the remaining seven recipients showed a significant trend of improvement . Combining the result of the seven patients reported previously, the improvement in terms of relaparotomy rate is significant . CONCLUSION: With modification of surgical technique, living donor liver transplantation in adults using right lobe liver grafts can become a relatively safe procedure. Eur J Pharmacol, 2000 Jan 3, 387(1), 1 - 7 The antibacterial and NMDA receptor activating properties of aminoglycosides are dissociable; Harvey SC et al.; The use of aminoglycoside antibiotics is limited by side effects, the most critical of which are vestibular and cochlear toxicity . Recent evidence indicates that these effects result from an excitotoxic process mediated, at least in part, through a polyamine-like activation of NMDA receptors . This study investigated whether these positive modulatory effects of aminoglycosides at NMDA receptors are dissociable from their antibacterial properties . A group of structurally related apramycin derivatives was evaluated for the ability to enhance {3H}dizocilpine binding to rat brain membranes, and for the ability to augment agonist responses on recombinant (NR1A/2B) NMDA receptors expressed in Xenopus oocytes . Based on the antibacterial potencies of these derivatives against Staphylococcus aureus and Escherichia coli, it is concluded that there is no correlation between the ability of an aminoglycoside to produce a positive modulation of NMDA receptors and minimum inhibitory antibacterial concentrations . These findings indicate that it may be possible to develop an aminoglycoside antibiotic with reduced potential for ototoxicity. J Hosp Infect, 2000 Jan, 44(1), 19 - 26 An epidemiological survey of methicillin-resistant Staphylococcus aureus in a tertiary referral hospital; Barakate MS et al.; Over a 30-month period from July 1995 to December 1997, new detections of methicillin-resistant Staphylococcus aureus (MRSA) were prospectively studied in a tertiary referral hospital . The aims of the study were to determine the incidence of colonization of patients admitted to each of the hospital's 39 clinical units and ascertain where each patient had become colonized . Epidemiological information (time to detection, ward movement, admission to other hospitals, data on MRSA isolations in hospital wards) and phage typing were used by the hospital's infection control unit to make this determination . Routine containment procedures included cohorting, flagging and triclosan body washes . Surveillance cultures were collected infrequently . Patients known to be colonized with MRSA were excluded from orthopaedic and haematology wards . During the study period, 995 patients were found to be newly colonized . The incidence of colonization varied from nil to 72 per 1000 admissions, being highest in the main intensive care unit and in services which frequently used that unit . The incidence of colonization in elective orthopaedic surgery (< 1 per 1000) and haematology (3 per 1000) was very low . Determining the place where patients acquired MRSA was made difficult by the high frequency of endemic phage types and frequent patient transfer between wards . Epidemiological data suggested that the main intensive care unit and surgical wards nursing patients with colorectal, urological and vascular diseases were the places where most patients became colonized . MRSA was never acquired by patients nursed in wards which practised an exclusion policy towards patients known to be colonized with MRSA . Our data suggest that in tertiary referral hospitals, where MRSA is not only endemic but frequently imported from other hospitals, it is possible to establish areas where MRSA is never acquired . Invest Ophthalmol Vis Sci, 2000 Jan, 41(1), 145 - 53 Adhesion molecule expression in a rat model of Staphylococcus aureus endophthalmitis; Giese MJ et al.; PURPOSE: To determine whether Staphylococcus aureus and its components induce expression of E-selectin and intercellular adhesion molecule (ICAM)-1 in rat ocular tissues and on human endothelial cells in culture . METHODS: Experimental and control rat eyes were injected with 80 colony-forming units of viable S . aureus and lipopolysaccharide-free sterile saline (NS), respectively . Eyes were enucleated and immediately frozen . E-selectin and ICAM-1 expression were evaluated on frozen sections by using standard immunohistochemical techniques . Using an enzyme-linked immunoassay, in vitro expression of E-selectin and ICAM-1 was evaluated on macrovascular endothelial cells after stimulation with S . aureus and selected purified components . RESULTS: In S . aureus-injected eyes, E-selectin and ICAM-1 expression peaked at six to 24 hours, decreased slightly at 24 and 48 hours, and further declined by 72 hours . However, in NS-injected eyes, peak levels of E-selectin and ICAM-1 were seen at 6 hours, after which expression declined in the areas in which an increase was previously observed . In in vitro assays, peptidoglycan (0.01 microg/ml) induced a fourfold increase in E-selectin (P < 0.0001) and a twofold increase in ICAM-1 (P < 0.002) expression . Ribitol teichoic acid (RTA) (1 microg/ml) induced a twofold increase in E-selectin (P < 0.0001) and a threefold increase in ICAM-1 (P < 0.0001) expression . CONCLUSIONS: Eyes injected with S . aureus demonstrated a more intense and prolonged expression of both E-selectin and ICAM-1 than did eyes injected with NS . In addition, S . aureus components induced the in vitro expression of these adhesion molecules on macrovascular endothelial cells . The relevance of these findings to microvascular endothelial cells is yet to be determined. J Bacteriol, 2000 Feb, 182(3), 664 - 71 Expression of the multidrug resistance transporter NorA from Staphylococcus aureus is modified by a two-component regulatory system; Fournier B et al.; To dissect genetically the regulation of NorA, a multidrug transporter of Staphylococcus aureus, we analyzed the differential expression of the norA promoter using a transcriptional fusion with a beta-lactamase reporter gene . Expression studies with an arlS mutant revealed that the norA promoter is ArlS dependent . The arlR-arlS locus was shown to code for a two-component regulatory system . The protein ArlR has strong similarity to response regulators, and ArlS has strong similarity to protein histidine kinases . We have also analyzed the 350-bp region upstream of the Shine-Dalgarno sequence of norA by gel mobility shift experiments . It was shown that only the 115-bp region upstream of the promoter was necessary for multiple binding of an 18-kDa protein . From transcriptional fusions, we have localized four different putative boxes of 6 bp, which appear to play a role in the binding of the 18-kDa protein and in the up-regulation of norA expression in the presence of the arlS mutation . Furthermore, the gel mobility shift of the 18-kDa protein was modified in the presence of the arlS mutation, and the arlS mutation altered the growth-phase regulation of NorA . These results indicate that expression of norA is modified by a two-component regulatory system. Comp Biochem Physiol B Biochem Mol Biol, 1999 Nov, 124(3), 281 - 8 Isolation and amino acid sequence of two trypsin isoinhibitors from duck pancreas; Wilimowska-Pelc A et al.; DPTI II and DPTI IV, two trypsin inhibitors from duck pancreas, have been isolated by affinity chromatography on immobilized anhydrotrypsin, anion exchange and RP-HPLC . The complete amino acid sequence of both inhibitors was determined after reductive carboxymethylation and digestion with Staphylococcus aureus V8 protease or trypsin . The inhibitors were each found to be a single polypeptide chain comprised of 69 amino acid residues and their molecular masses were estimated at 7687 Da for DPTI II and 7668 Da for DPTI IV . The only difference in amino acid sequence between the two inhibitors is the replacement of Arg for His residue in the C-terminal position of DPTI IV. J Orthop Res, 1999 Nov, 17(6), 947 - 52 Contamination of the medullary canal following pin-tract infection; Clasper JC et al.; We developed an ovine model of an external-fixator pin-tract infection . With use of a novel method of tissue sampling, infection of the medullary canal was confirmed in all (10 of 10) external-fixator pins that were contaminated with Staphylococcus aureus after they were inserted . In addition, all (five of five) adjacent, uncontaminated pins became infected . We demonstrated that pin-tract infection can be difficult to diagnose clinically, despite gross infection of the tract, and that bacteria can spread within the medulla . Three of the infected pins (20%) did not appear clinically infected, were not loose, and were normal on radiographs after 1 week . Staphylococcus aureus was isolated from the medulla around all 15 pin tracts, and nine other organisms were isolated from the tracts . Despite the presence of infection, the majority of the pins remained well fixed in the bone after 2 weeks. J Chemother, 1999 Oct, 11(5), 331 - 7 Emergence of resistance during mupirocin treatment: is it a problem in clinical practice? Henkel T, Finlay J. Mupirocin (pseudomonic acid A) is indicated for primary and secondary skin infections, and for the eradication of nasal colonization of Staphylococcus aureus, particularly methicillin-resistant S . aureus (MRSA) . This paper reviews the mechanisms by which resistance to mupirocin can develop, discusses clinically relevant breakpoints, and the clinical significance of reports of resistance . Following more than 10 years' use, short courses of treatment, even when repeated, are associated with remarkably little resistance and this resistance is unlikely to be clinically significant. Gynakol Geburtshilfliche Rundsch, 1999, 39(4), 217 - 25 {Experimental investigations and clinical use of photodynamic therapy (PDT) in the Rudolf Foundation Hospital}; Wierrani F; This article addresses experimental investigations and the clinical use of PDT in the Rudolfstiftung Hospital, Vienna . We investigated mesotetrahydroxyphenylchlorine (mTHPC) and the photosensitizer hematoporphyrin derivative alone or in combination to prove photodynamic antibacterial effects on Staphylococcus aureus (wild type) . mTHPC showed antibacterial toxicity in the dark; hematoporphyrin derivative showed suppressive growth effects only after white-light illumination . Photodynamic activity by the combination of both dyes was obtained in a roughly additive manner . Furthermore, we observed the development of resistance of erythromycin after the illumination procedure with hematoporphyrin derivative . Wild-type S . aureus developed no resistance to the other antibiotics tested . Furthermore, long-term follow-up examinations proved mTHPC-mediated PDT as a possible adjuvant intraoperative therapy in cases of relapses of gynecologic carcinomas . PDT is a tissue-selective and simple intervention . It shows few side effects, and, therefore, it reduces the overall burden of tumor patients . In another clinical investigation, we used 5-aminolevulinic acid-based PDT to treat intraepithelial neoplasia and human papillomavirus of the uterine cervix . 33 of 38 (86,8%) patients with superficial cervical intraepithelial neoplasia grades I and II were treated successfully with PDT . Eradication of human papillomavirus infections was successfully performed in 80% of the cases . Antibiot Khimioter, 1999, 44(11), 34 - 6 {The role of cefepime, a 4th-generation cephalosporin, in treating patients with surgical sepsis}; Shliapnikov SA et al.; Cefepime (Maxipime) was used in the management of 22 patients at the age of 18 to 73 years with the surgical sepsis syndrome (SAPS > 15) . In 16 patients surgical sepsis was due to pancreatitis, appendititis, abdominal wound or trauma or complications after planned surgical interventions on the organs of the abdominal cavity . In the other 6 patients surgical sepsis was due to inflammatory processes in soft tissues after minor trauma . In 10 patients (group 1) cefepime was used after the pathogen verification and antibioticogram examination . In 12 patients (group 2) the antibiotic was used in the empirical therapy as the first line drug after the patients acceptance from another unit when the pathogen nature was obscure . Cefepime was administered intravenously in a dose of 2.0 g twice daily for 7 to 10 days in combination with metronidazole in a dose of 0.5 g thrice daily . After 5-6 days of the treatment the patients of group 1 were switched to the cefepime intramuscular regimen . The lethality totaled 18 per cent (4 patients) . Three of them were from group 2 . The patients died of progressive polyorgan insufficiency . It is characteristic that in no cases cefepime induced septic shock due to the endotoxin escape . No septicopyemia was as well observed even in the patients with verified bacteremia due to Staphylococcus aureus. J Food Prot, 2000 Dec, 63(12), 1713 - 8 Evaluation of batch and semicontinuous application of high hydrostatic pressure on foodborne pathogens in salsa; Raghubeer EV et al.; The effects of high hydrostatic pressure (HPP; 545 MPa) on strains of Escherichia coli O157:H7, Listeria monocytogenes, enterotoxigenic Staphylococcus aureus, and nonpathogenic microorganisms were studied in tomato-based salsa . Products were evaluated for the survival of the inoculated pathogens following HPP treatment and after storage at 4 degrees C and 21 to 23 degrees C for up to 2 months . Inoculated samples without HPP treatment, stored under the same conditions, were also evaluated to determine the effects of the acid environment of salsa on the survival of inoculated strains . None of the inoculated pathogens were detected in the HPP-treated samples for all treatments throughout the storage period . Inoculated pathogens were detected in the non-HPP-treated samples stored at 4 degrees C after 1 month, with L . monocytogenes showing the highest level of survivors . In the non-HPP-treated samples stored at 21 to 23 degrees C, E . coli and S . aureus were not detected after 1 week, but L . monocytogenes was detected in low levels . Studies with nonpathogenic strains of the pathogens were conducted at Oregon State University using HPP treatments in a semicontinuous production system . The nonpathogenic microorganisms (E . coli, Listeria innocua, Listeria welshimeri, and nonenterotoxigenic S . aureus) were inoculated together into a feeder tank containing 100 liters of salsa . Microbiological results of samples collected before HPP treatment and from the aseptic filler were similar to those obtained for the pathogenic strains . No survivors were detected in any of the HPP-treated samples. Pediatrics . 2000 Dec;106(6):E87. The pivotal role of deep vein thrombophlebitis in the development of acute disseminated staphylococcal disease in children; Gorenstein A et al.; Deep vein thrombophlebitis (DVT) and septic pulmonary emboli (PE) are rare in children . The association of DVT and acute disseminated staphylococcal disease (DSD) during childhood has not been previously reported . We report 3 children who developed a triad of DVT, septic PE, and acute osteomyelitis with Staphylococcus aureus cultured from blood and bone . One child succumbed, while 2 survived following prolonged, morbid hospitalizations . The rapid clinical deterioration observed in these patients might be caused by the aggressiveness of staphylococcal infection combined with an ongoing showering of septic emboli from the ileo-femoral DVT . We suggest that infected DVT with septic PE had a pivotal role in the development of DSD in these children . The presence of this triad should prompt aggressive treatment with the appropriate antibiotics, anticoagulation, surgical drainage, and assisted ventilation when indicated. Clin Microbiol Rev, 2000 Jan, 13(1), 16 - 34, table of contents Exotoxins of Staphylococcus aureus; Dinges MM et al.; This article reviews the literature regarding the structure and function of two types of exotoxins expressed by Staphylococcus aureus, pyrogenic toxin superantigens (PTSAgs) and hemolysins . The molecular basis of PTSAg toxicity is presented in the context of two diseases known to be caused by these exotoxins: toxic shock syndrome and staphylococcal food poisoning . The family of staphylococcal PTSAgs presently includes toxic shock syndrome toxin-1 (TSST-1) and most of the staphylococcal enterotoxins (SEs) (SEA, SEB, SEC, SED, SEE, SEG, and SEH) . As the name implies, the PTSAgs are multifunctional proteins that invariably exhibit lethal activity, pyrogenicity, superantigenicity, and the capacity to induce lethal hypersensitivity to endotoxin . Other properties exhibited by one or more staphylococcal PTSAgs include emetic activity (SEs) and penetration across mucosal barriers (TSST-1) . A detailed review of the molecular mechanisms underlying the toxicity of the staphylococcal hemolysins is also presented. Plast Reconstr Surg, 2000 Jan, 105(1), 334 - 8; discussion 339-43 Optimizing breast pocket irrigation: an in vitro study and clinical implications; Adams WP Jr et al.; Subclinical infections have been implicated in the etiology of capsular contracture . Intraoperatively, breast pocket irrigation with povidone-iodine or other antibiotic solutions has been popularized; however, detrimental effects on wound healing for these agents have been reported and their efficacy against common organisms found around breast implants has not been studied . The purpose of this study was to compare the in vitro efficacy of serial dilutions of povidone-iodine and two double antibiotic solutions DAB-1 (gentamicin/polymyxin B) and DAB-2 (gentamicin/cefazolin), against organisms most commonly found around breast implants . In phase I trials, serial dilutions of povidone-iodine and DAB were combined 1:1 with cultures of five common organisms found around implants . In phase II, povidone-iodine was serially diluted in DAB-1 rather than saline . In phase III, povidone-iodine was serially diluted with DAB-2 . Efficacy for all phases was determined by plating the mixture onto agar plates and incubating at 37 degrees C for 48 hours . Povidone-iodine was 100 percent effective at a dilution of 12.5% against Staphylococcus epidermidis and 25% against Staphylococcus aureus but relatively ineffective against Escherichia coli and Pseudomonas, DAB-1 was found to be ineffective against S . epidermidis but effective against S . aureus, Propionibacterium acnes, E . coli, and Pseudomonas . In phase II trials, a concentration of 12.5% povidone-iodine in DAB was effective at killing all experimental bacteria . In phase III trials, 10% povidone-iodine in DAB-2 was effective at killing all bacteria tested . In conclusion, to maximize bacterial control of common breast implant organisms and to minimize the detrimental effects on wound healing, 10% povidone-iodine in gentamycin/cefazolin may be used with excellent results and its use clinically may reduce the incidence of capsular contracture. Poult Sci, 1999 Dec, 78(12), 1711 - 6 Delayed-type hypersensitivity reaction induced in broilers via trachea inoculation of killed Staphylococcus aureus; Zhu XY et al.; A study was conducted to determine whether the delayed-type hypersensitivity (DTH) reaction to killed Staphylococcus aureus antigen in chickens could be induced through multiple intratracheal inoculations . Three criteria were used to assess DTH: 1) delayed footpad reaction (DFR) with a peak response at 24 to 48 h postchallenge, 2) inhibition of monocyte/macrophage migration, and 3) mononuclear cell infiltration at the challenge site . Broilers were sensitized three times with a s.c . injection in the neck or intratracheal inoculation of killed S . aureus in polyethylene glycol at 2, 3, and 4 wk of age . Controls were given polyethylene glycol with a s.c . injection in the neck or intratracheal inoculation . Migration inhibition tests were conducted at 6 wk of age . At 7 wk of age, all birds were challenged intradermally with S . aureus antigen in PBS in the right footpad . The left footpad was injected with PBS . The thickness of the footpad was measured at 0, 4, 24, and 48 h postchallenge to evaluate the DFR . Birds were euthanatized, and both footpads were removed for histopathological examination . Subcutaneously or intratracheally sensitized birds showed significant DFR compared with nonsensitized birds (P < 0.0001), which reached maximum response at 24 h postchallenge . The s.c . sensitization resulted in an inhibition of the in vitro migration of monocytes/macrophages (P < 0.0001), whereas intratracheally sensitized birds did not show migration inhibition of monocytes/macrophages . Histological examination showed typical perivascular infiltration of small lymphocytes in S . aureus-injected footpads from s.c . and intratracheally sensitized birds . These results indicate that multiple intratracheal inoculation, as well as s.c . injection of killed S . aureus antigen, can be used to induce a cell-mediated DTH reaction in chickens. Poult Sci, 1999 Dec, 78(12), 1703 - 10 Delayed-type hypersensitivity reaction induced in broilers by killed Staphylococcus aureus; Zhu XY et al.; A trial was conducted to determine whether the delayed footpad reaction (DFR) induced by killed Staphylococcus aureus in chickens is a delayed-type hypersensitivity (DTH) reaction . Five criteria were used to assess DTH: 1) DFR with a peak response at 24 to 48 h postchallenge, 2) inhibition of monocyte/macrophage migration, 3) lymphocyte blastogenic response, 4) mononuclear cell infiltration at the challenge site, and 5) passive transfer of DFR by splenic lymphocytes . Broilers were sensitized twice with a s.c . injection in the neck of S . aureus antigen (150 microg/bird) diluted in polyethylene glycol at 3 and 4 wk of age . Controls were s.c . injected with polyethylene glycol . At 6 wk of age, a migration inhibition test was conducted before the birds were challenged intradermally with S . aureus antigen (75 microg/bird) in PBS in the right footpad . The left footpad was injected with PBS . The thickness of the footpad was measured at 0, 4, 24, and 48 h postchallenge to evaluate the DFR . After challenge, blood was collected for the lymphocyte blastogenesis assay . Birds were euthanatized, and both footpads were removed for histology . The spleens were collected aseptically; splenic lymphocytes were injected i.v . into recipient birds . Sensitized birds showed an increase in the DFR (P < 0.02) and blastogenic response (P < 0.01) compared with nonsensitized birds . Delayed footpad reaction reached a maximum response at 24 h postchallenge . The in vitro migration of monocytes/macrophages from sensitized birds was significantly inhibited (P < 0.01) . The histological appearance of S . aureus-injected footpads was characterized by dermal edema and perivascular infiltrates of small lymphocytes and macrophages . Birds that received sensitized splenic lymphocytes had a significantly pronounced DFR following challenge with S . aureus when compared with birds that received nonsensitized lymphocytes (P < 0.0001) . These results indicated that the DFR can be used as a standard in vivo test for cell-mediated DTH reaction induced by killed S . aureus antigen in chickens. Biochemistry, 2000 Jan 11, 39(1), 26 - 36 Characterization of the binding interface between the E-domain of Staphylococcal protein A and an antibody Fv-fragment; Meininger DP et al.; Staphylococcal protein A (SpA) is a cell-surface component of Staphylococcus aureus . In addition to the well-characterized interaction between SpA and the Fc-region of human IgG, an alternative binding interaction between SpA and the Fab-region of immunoglobulin domains encoded by the V(H)3 gene family has been described . To characterize structurally the interface formed by SpA repeats and type-3 V(H)-domains, we have studied the 32-kDa complex formed between an E-domain mutant (EZ4) and the Fv-fragment of the humanized anti-HER2 antibody (Hu4D5-8) using heteronuclear NMR spectroscopy . Protocols were established for efficient incorporation of (15)N, (13)C, and (2)H into EZ4 and the V(H)- and V(L)-domains of the Fv, allowing backbone resonances to be assigned sequentially for EZ4 and the V(H)-domain in both free and complexed states . Broadening of certain V(H)-resonances in the free and bound Fv-fragment suggests microsecond to millisecond time-scale motion in CDR3 . Residues experiencing significant chemical shift changes of backbone (1)H(N), (15)N, and (13)CO resonances upon complex formation delineate contiguous surfaces on EZ4 and the V(H)-domain that define the binding interfaces of the two proteins . The interaction surfaces identified by chemical shift mapping are comprised