J Infect Dis, 1996 Oct, 174(4), 777 - 85 Differential activation of extracellular signal-regulated kinase (ERK) 1, ERK2, p38, and c-Jun NH2-terminal kinase mitogen-activated protein kinases by bacterial peptidoglycan; Dziarski R et al.; Soluble staphylococcal peptidoglycan (sPGN) is an inducer of cytokine secretion and may activate macrophages through the CD14 lipopolysaccharide (LPS) receptor . To elucidate sPGN-activated signal transduction pathways, stimulation of mitogen-activated protein (MAP) kinases by sPGN was studied in mouse RAW264.7 macrophages . sPGN strongly activated extracellular signal-regulated kinase (ERK) 1 and ERK2, moderately activated c-Jun NH2 terminal kinase (JNK), and weakly activated p38 MAP kinase, in contrast to LPS, which strongly activated all of these kinases, and phorbol 12,13-dibutyrate (PDB), which strongly activated ERK1 and ERK2 but did not activate p38 or JNK . sPGN- and LPS-induced activation of ERK1 and ERK2, unlike PDB-induced activation, was sensitive to inhibition by herbimycin A and insensitive to inhibition by increased intracellular cAMP . These results demonstrate differential activation of MAP kinases by sPGN, similar but not identical activation of signal transduction pathways by sPGN and LPS, and different mechanisms of MAP kinase activation by bacterial stimulants and phorbol esters. J Bacteriol, 1996 Oct, 178(20), 6070 - 3 IS257-mediated cointegration in the evolution of a family of staphylococcal trimethoprim resistance plasmids; Leelaporn A et al.; Analyses of the Staphylococcus epidermidis multiresistance plasmids pSK697 and pSK818 have revealed them to be closely related to the trimethoprim resistance plasmid pSK639, also isolated from S . epidermidis . pSK697 and pSK818 were found to contain a cointegrated copy of a second plasmid related to the S . epidermidis multidrug antiseptic and disinfectant resistance plasmid pSK108 and the S . aureus tetracycline resistance plasmid pT181, respectively . In contrast to pSK639, both plasmids were found to contain a third copy of IS257, such that the integrated plasmids in both cases are flanked by a copy of this element . This organization and the presence of duplicated sequences at the extremities of the integrated plasmids implicate IS257 in the formation of these cointegrate plasmids . Sequence analysis of the IS257 elements from these plasmids has provided insights into the probable mechanism of cointegration, viz., nonresolved replicative transposition of IS257. J Immunol, 1996 Oct 1, 157(7), 2982 - 8 Staphylococcal protein A simultaneously interacts with framework region 1, complementarity-determining region 2, and framework region 3 on human VH3-encoded Igs; Potter KN et al.; Staphylococcal protein A (SPA) is a B cell superantigen that binds to human VH3-encoded Igs independently of the D- and JH-encoded regions and light chain sequences . The SPA-binding structure formed by VH3-encoded Igs remains controversial . We localized the regions in a VH3-encoded Ab required for SPA binding by producing mutant Abs in the baculovirus expression system in which regions of a human-derived Ab known to bind SPA were exchanged with those from a mouse Ab of the J558 family, a family not associated with SPA binding . The pattern of SPA binding indicates not only that residues in FR1, CDR2, and FR3 are involved but also that the three regions are required to interact simultaneously with SPA for binding to occur . When any one of the three regions was replaced with the corresponding region from the nonbinding Ab, SPA binding was severely disrupted . These data indicate that SPA requires simultaneous interaction with three distinct regions of a VH3 structure, which together in three-dimensional space form an extended solvent-exposed surface . These studies more precisely define the genetic requirements for VH3-encoded Ig binding to SPA. J Immunol, 1996 Oct 1, 157(7), 2976 - 81 Differential binding avidities of human IgM for staphylococcal protein A derive from specific germ-line VH3 gene usage; Hakoda M et al.; Human IgM that express the variable region genes of the VH3 family bind staphylococcal protein A (SPA) . We previously reported that the SPA-binding IgM can be divided into two groups based on the differential binding avidities for solid-phase SPA . To study the molecular basis for these differences, we cloned B cells from human blood by EBV transformation . The nucleotide sequences of the expressed Ig heavy chain genes were determined on 20 B cell clones that produce SPA-binding IgM . The germ-line VH3 gene usage in IgM with high avidities for SPA were distinct from the germ-line VH3 genes used in IgM with low avidities for SPA . There was no correlation in the usage of D or JH genes or in the usage of light chains in IgM according to the SPA binding avidity . These results suggest that the differential binding avidities for SPA are at least partly due to specific germ-line VH3 gene usage . An investigation of direct binding of SPA to the synthetic peptides corresponding to the portions of the variable regions of SPA-binding and non-SPA-binding IgM showed that the peptides corresponding to the VH3 family specific framework region 3 sequences had significant SPA binding capacities, while the peptides corresponding to the other subdomains and those corresponding to framework region 3 of the reported VH3 sequences from non-SPA-binding IgM showed little or no binding . It is of interest that the Ig-framework region 3 subdomain corresponds to the fourth hypervariable region, which in the TCR-beta chain has been implicated as a critical site for T cell superantigen binding. J Immunol, 1996 Oct 1, 157(7), 2857 - 63 Regulation of superantigen-induced T cell activation in the absence and the presence of MHC class II; Lando PA et al.; To study MHC class II-dependent and -independent SAg2 activation and the relative importance of CD80/CD28 costimulation, staphylococcal enterotoxin A (SEA) was presented to T cells as a fusion protein containing the Fab fragment of an mAb directed against the CA215 glycoprotein . Chinese hamster ovary (CHO) cells transfected with HLA-DR4, CA215, and CD80, individually or in combinations, were used as presenting cells . A strong T cell proliferation was obtained when C215Fab-SEA fusion proteins were presented by CHO-DR/CD80 or CHO-CA215/CD80 double transfectants, whereas only low levels of proliferation were seen in the absence of CD80 . Large amounts of IL-2, IFN-gamma, and TNF were produced in addition to an increase in IL-2 mRNA as a result of CD80 costimulation . Only approximately 50% of the SEA-reactive T cells responded by expression of IL-2 receptor chains and by blast formation when activated with SEA in the absence of MHC class II . Reverse transcription-PCR-assisted repertoire analysis of SEA-reactive TCR V beta families showed that the CA215-dependent activation involved an expansion of fewer TCR V beta families compared with MHC class II-dependent activation . One-half of the six analyzed TCR V beta families were expanded independently of class II . This indicates that MHC class II has only a partial influence on the TCR V beta repertoire imprinted by SAg . This finding redefines the role of MHC class II in SAg presentation . It is suggested that MHC class II molecules are selected as SAg-binding molecules mainly as a suitable targeting receptor for professional APC expressing costimulatory molecules such as CD80 and CD86. Presse Med, 1996 Sep 28, 25(28), 1276 - 80 {Outcome of cardiac valve ring abscesses after medical treatment: attempt to identify criteria of favorable prognosis}; Tribouilloy C et al.; OBJECTIVES: Identify factors predicting favorable outcome after medical management of valve ring abscesses in order to propose a surveillance schedule for conservative treatment . METHODS: A multicentric study conducted from July 1989 to February 1996 included 28 patients (mean age 64 +/- 16 years, range 26-83) hospitalized for active endocarditis and valve ring abscesses diagnosed at transthoracic or transesophageal echography . Conservative medical therapy was given because of a decision of the medico-surgical team (n = 9), high surgical risk (n = 12), or patient refusal of surgery (n = 7) . Outcome was favourable in 18 patients (Group I) and unfavorable in 10 (Group II) due to death (n = 9) or subsequent surgery (n = 1) . Univariate and multivariate analysis were used to determine differences between the groups in terms of clinical and laboratory data . RESULTS: Mean follow-up in Group I was 33 +/- 18 months and 15 +/- 10 months in Group II . Univariate analysis showed significant differences between Group I and II respectively for age (59 +/- 18 yr vs 72 +/- 10, p = 0.04), delay to apyrexia after antibiotics (4.3 +/- 2.8 vs 8.3 +/- 2.4 days, p < 0.0008), heart failure (5% vs 70%, p = 0.003), grade III or IV valvular regurgitation (5% vs 60%, p < 0.04), and mean surface area of the abscess (1.5 +/- 1.2 vs 5.4 +/- 6.4 cm2, p < 0.03) . Independent factors at multivariate analysis were by decreasing order: lack of heart failure at admission, delay to apyrexia, abscess surface area, and age . Outcome was favorable (mean follow-up 33 +/- 10 months) in all patients with an abscess surface area < 1.5 cm2, no signs of heart failure, no grade III or IV valvular regurgitation, apyrexia after less than 8 days on antibiotics and no staphylococcus positive blood culture . CONCLUSION: Medical management of valve ring abscesses may be indicated in selected patients in care units with rigorous surveillance facilities . Further studies are needed to precisely identify surveillance and treatment criteria. Int J Cancer, 1996 Sep 27, 68(1), 109 - 13 Immune response during tumor therapy with antibody-superantigen fusion proteins; Rosendahl A et al.; To engineer superantigens (SAg) to express tumor reactivity, we genetically fused the Fab-part of the tumor-reactive MAb C215 and the bacterial SAg staphylococcal enterotoxin A (SEA) . Treatment of mice carrying established lung micrometastases of the C215-transfected syngeneic B16 melanoma with 3-4 daily injections of C215Fab-SEA resulted in strong antitumor effects, while only moderate effects were seen when treatment was given every 4th day (intermittent treatment) . High serum levels of IL-2, TNF-alpha, IFN-gamma and strong induction of CTLs (cytotoxic T lymphocytes) were noted after priming with the fusion protein . T cells responded well to 3 daily injections of C215Fab-SEA and then gradually entered a hyporesponsive state, characterized by a reduced ability to produce IL-2, TNF-alpha and IFN-gamma and failure to mediate cytotoxicity in vitro . Intermittent treatment was characterized by increased levels of IL-10, concomitant with accentuated loss of IL-2, TNF-alpha and IFN-gamma production . A 10-fold increase in SEA-reactive TCR V(beta)3+ CD4+ cells was observed in the spleen, while a loss of TCR V(beta)3+ CD8+ and V(beta)11+ CD8+ cells was noted . This is in striking contrast to injections of native SEA which induced a marked deletion of TCR V(beta)3+ CD4+ T cells, but not of CD8+ cells . Recovery of the TH1 cytokine profile occurred within 1-2 weeks, while restoration of cytotoxicity required several months and correlated with recovery of TCR V(beta)3+ CD8+ and TCR V(beta)11+ CD8+ T cells . These results show that the temporal relationship of SAg stimulations dictates the cytokine profile . Moreover, different mechanisms appear to regulate hyporesponsiveness in CD4+ and CD8+ T cells. Cell Immunol, 1996 Sep 15, 172(2), 254 - 61 The immunodominant region of Staphylococcal nuclease is represented by multiple peptide sequences; Nikcevich KM et al.; Several published reports have lead to the characterization of naturally processed peptides that are presented in association with either class I or class II MHC molecules . Most peptides isolated from class II molecules are heterogeneous in length and exhibit ragged amino and carboxy termini . An intriguing finding was that one region of a molecule was often represented by many distinct peptides, rather than by a single dominant peptide species . Each of the peptides representing this dominant region exhibited a common core of amino acids, suggesting that this core may play a significant role in the binding of the peptide to class II and the recognition by peptide-specific T cells . Work from our laboratory has focused on the mechanisms involved in the immunodominance of antigenic determinants using the bacterial antigen Staphylococcal nuclease (Nase) as a model . Using truncated synthetic peptides, we have identified the immunodominant determinant of Nase to be located within the region 81-100 with a minimal antigenic core of 91-100 as determined . Addition of five residues to the carboxy terminus of this peptide had a negative effect on T cell recognition of this region . The present studies were undertaken in an effort to determine the sequence of the naturally processed immunodominant Nase determinant(s) presented in association with I-Ek class II . Our results indicate that the dominant region of the Nase molecule is represented by at least four distinct peptide species that are predicted to lie between residues 86 and 106 with a common core sequence of 91-96 . These results indicate that the negative effects of flanking regions are dependent upon length and amino acid composition, and thus the use of truncated peptides to study minimal antigenic determinants may be misleading. Minerva Chir, 1996 Sep, 51(9), 707 - 11 {A rare case of primary abdominal actinomycosis}; Lazzaretti MG et al.; The authors describe a case of primary abdominal actinomycosis operated on because of peritonitis sustained by a tubo-ovarian abscess . They discuss the pathogenesis of the case: the patient had been on intrauterine device contraception till two months earlier and had been operated on for breast cancer . Preoperative diagnosis is quite impossible and only the microscopic observation of the specimen can show the causative agent . Surgical options are reported, stressing the need for an adequate period of antimicrobial therapyPIP: In September 1993 a 43-year-old female patient with cancer underwent left mastectomy followed by immediate reconstruction . 6 days passed without problems, but then she presented at the emergency ward with abundant exudation of serous material from the cicatrices . Microbiological test showed evidence of Staphylococcus epidermitis . Drainage of the skin and smooth muscle was performed and the secretion was immediately reduced and seemed to disappear in a short time . In the next 3 days fever arose accompanied by abdominal pain . Blood test showed leucocytosis (24,500 GB), increase of the suppressor lymphocytes (CD8) and the reduction of CD4/CD8 ratio . Abdominal-pelvic echogram showed evidence of an enlarged right adnexum as well as that of the homolateral tube, but no discharge of fluid in the pelvic cavity . Gynecological examination in this patient, who had worn an IUD two months prior, excluded lesions in the portio or vagina and the vaginal flora did not show fungi or parasites . Diagnostic laparoscopy followed, which demonstrated in the pelvic cavity a large para-uterine tumefaction . The pelvic organs were adhering to the parietal layer of the peritoneum and in the whole peritoneal cavity, including the interhepatic-diaphragmatic space, fibrin plaque and pus was observed . Laparotomy was performed, which confirmed a parauterine mass and a tubo-ovarian complex with numerous recesses containing fetid, grayish pus . Complete right adnexectomy was carried out with abundant lavage and multiple drainage of the peritoneal cavity . Subsequently, the abdominal situation improved, but a new examination of drained liquid showed the presence of cutaneous bacterial flora but no fungi or parasites . Ovarian actinomycotic abscess with acute peritonitis and salpingitis was demonstrated . Subsequent antibiotic therapy consisted of piperacilline for 15 days, and 4 months after the episode the patient was well without return of the foci of infection . Diagn Microbiol Infect Dis, 1996 Sep, 26(1), 43 - 5 The utility of non-beta-lactam antimicrobial MICs as markers to distinguish oxacillin-resistant from oxacillin-susceptible strains of Staphylococcus epidermidis; Fass RJ et al.; Among 6,068 strains of Staphylococcus epidermidis, 75.5% were oxacillin-resistant . Oxacillin-susceptible strains were more frequently susceptible to erythromycin, clindamycin, ciprofloxacin, trimethoprim/sulfamethoxazole, gentamicin, and tetracycline than oxacillin-resistant strains . With the exception of erythromycin, non-beta-lactam MICs were less discriminatory for identifying oxacillin-resistant strains with oxacillin MICs < or = 2 micrograms/ml than for those with oxacillin MICs > or = 4 micrograms/ml. ASAIO J, 1996 Sep-Oct, 42(5), M881 - 4 Evidence that bacteria prefer to adhere to thrombus; Bos HM et al.; This study was undertaken to investigate the possible association between thrombosis and infection using an in vitro test model in which fresh bovine blood was recirculated through test conduits (3.5 mm inner diameter) containing stent-like devices . Anticoagulation was adjusted so that the recirculating blood deposited thrombi on the stent to cause gradual occlusion, thus impeding the flow . Four stent-like devices were placed in separate conduits in each experiment, and blood was recirculated with the help of pneumatically driven ventricles . Flow through these conduits was monitored by ultrasonic flow detection . To quantitate bacterial interaction with thrombi, Staphylococcus epidermidis (15E10(9)) was labeled with 111Indium-oxine and added to the blood . Experiments lasted until the flow in the test conduits dropped to 10% of the starting flow . During this recirculation, as flow gradually decreased, one stent was taken out when flow was still at 100%, the second at 75%, the third at 50%, and the fourth at 10% of the starting flow . The number of bacteria associated with the thrombus was measured by gamma counting . The following observations were made: 1) the amount of thrombus increased with time in all experiments (this was confirmed in separate experiments by using autologous 111Indium labeled platelets); 2) bacterial adhesion showed a concomitant increase as thrombus size increased (this was confirmed by using 111Indium labeled bacteria), and 3) bacterial incorporation into the thrombus occurred regardless of whether they were viable or pretreated with the antibiotic rifampin . These observations suggest that as thrombi develop, they may preferentially attract micro-organisms . This suggests that devices with adherent thrombi may have greater susceptibility for infection. ASAIO J, 1996 Sep-Oct, 42(5), M645 - 8 Immunoadsorption with protein A in humoral rejection of kidney transplants; Pretagostini R et al.; The presence of alloantibodies may play a role in accelerated or acute humoral rejection . Different therapeutic strategies based on a removal of anti donor antibodies and prevention of their resynthesis have been used in the management of transplant rejection episodes . Immunoadsorption with staphylococcal protein A, a method to selectively remove immunoglobulin G, may represent a new treatment to reverse humoral rejection in kidney transplantation . From 1991 to January 1996, such a method was used in 23 patients in whom an acute humoral rejection developed over a mean period of 14.1 +/- 9.5 days after operation . Twenty-two patients had been transplanted from living donors and one from a cadaveric donor . The ages ranged from 23 to 58 years (mean, 34 +/- 10 years) . All transplants were performed according to a negative direct crossmatch . Basic immunosuppression included cyclosporine, steroids, azathioprine, and antilymphocyte globulin or monoclonal antibodies (OKT3) . Rejection was diagnosed on the basis of hematochemical tests, Doppler ultrasonography, and kidney biopsy . Only steroid and monoclonal and polyclonal antibody resistant rejections with > 165% positive direct crossmatches against the donor were treated with Protein A immunoabsorption . The procedure used is based on the treatment of 2-3 plasma volumes for the first 2 days and then every other day until a negative crossmatch is obtained, together with improvement in clinical status (mean treatments, 7.3 +/- 4.5 {range, 4-23}; mean duration of treatment, 12.3 +/- 10.2 days {range, 3-44}) . From the start of treatment, azathioprine is replaced by cyclophosphamide at a dose of 1-2 mg/kg/day . During treatment, a remarkable fall in immunoglobulin G levels is achieved on the first day, whereas immunoglobulin M titers remain constant, with a slight decrease in serum albumin . Immediately after treatment, a negative crossmatch was found in 22 (95.6%) of 23 patients . In six patients (26%), graft function did not recover, and one patient (4.3%) died . Preliminary results show that immunoabsorption with staphylococcal protein A may be an effective support in the treatment of humoral acute rejection, particularly when it is performed as soon as an early diagnosis of humoral rejection is made . In fact, such treatment has a highly selective adsorption, allows treatment of large volumes of plasma, and can achieve a rapid decrease in the titer of circulating immunoglobulins. Toxicol Pathol, 1996 Sep-Oct, 24(5), 619 - 26 Potentiation of inhaled staphylococcal enterotoxin B-induced toxicity by lipopolysaccharide in mice; LeClaire RD et al.; Nonhuman primates are the established model for evaluating toxic responses to staphylococcal enterotoxins (SEs), as they react similarly to humans . Rodents are generally considered unresponsive to SEs . Binding affinities and T-cell reactivity suggest that SE binds more efficiently to primate major histocompatability complex class II receptors than to mouse receptors . We investigated the potentiation of staphylococcal enterotoxin B (SEB) inhalation toxicity by lipopolysaccharide (LPS) in BALB/c mice . Lethality occurred only when SEB was potentiated by LPS . Neither SEB nor LPS produced lethal effects alone . Temporal responses of interleukin 1 alpha, tumor necrosis factor alpha, interleukin 2, and interferon-gamma evoked by inhaled SEB were enhanced by LPS . By 24 hr after intoxication, serum cytokines decreased to baseline levels, and consistent pulmonary perivascular leukocytic infiltrates were evident histologically . Histologic lesions induced by inhalation exposure to SEB by mice, with or without potentiation by LPS, were similar to those in the rhesus monkey . Predominant pulmonary lesions included severe, diffuse interstitial and alveolar pulmonary edema, leukocytic infiltrates, mild perivascular edema, and alveolar fibrin deposition . Although the mechanism of aerosolized SEB-induced toxicity has not been completely resolved, similarities in histologic lesions, cytokine responses, and acute dose-response suggest the LPS-potentiated mouse model may be a credible alternative to the nonhuman primate model. J Perinatol, 1996 Sep-Oct, 16(5), 331 - 5 Vancomycin cerebrospinal fluid concentrations after intravenous administration in premature infants; Reiter PD et al.; OBJECTIVE: Staphylococcal species are the most common cause of nosocomial infections in the neonate . Because of staphylococcal resistance patterns, vancomycin has become the drug of choice for treatment . Although the blood stream is the usual site of infection, premature infants are at increased risk for the development of meningitis . The aim of this study was to determine vancomycin cerebrospinal fluid (CSF) concentration and penetration following intravenous (IV) administration in critically ill premature infants . STUDY DESIGN: A multiple-dose, open-label, case series was performed at a level III neonatal intensive care unit in a university teaching hospital . Three critically ill premature infants, 26 to 31 weeks of gestation requiring a course of IV vancomycin for suspected or proved sepsis were studied . Vancomycin was administered intravenously at 20 mg/kg, every 18 to 24 hours over 60 minutes . Serum and CSF vancomycin concentrations were obtained and pharmacokinetic analysis and CSF penetration was calculated . RESULTS: Serum vancomycin pharmacokinetics were consistent with those previously reported . CSF vancomycin concentrations ranged from 2.2 to 5.6 micrograms/ml and the calculated vancomycin CSF penetration ranged from 26% to 68% . CONCLUSIONS: CSF penetration of vancomycin after IV administration was much higher than that reported in older infants and children . This higher penetration may improve clinical outcomes in neonates with central nervous system infections . These data should be encouraging to clinicians who choose to use IV vancomycin for neonatal meningitis. Perit Dial Int, 1996 Sep-Oct, 16(5), 505 - 10 Continuous peritoneal dialysis-associated peritonitis of nosocomial origin; Troidle L et al.; OBJECTIVE: To describe our experience with nosocomial continuous peritoneal dialysis (CPD)-associated peritonitis focusing on the incidence, possible risk factors, spectrum of organisms, and outcome . DESIGN: Retrospective review of the medical records of our CPD patients admitted to an acute-care hospital between November, 1993 and December, 1994 . SETTING: University-associated acute-care hospitals in New Haven, Connecticut . PATIENTS: One hundred and eighty-eight patients maintained on CPD therapy and admitted to an acute-care hospital . RESULTS: Nineteen patients (5%) developing nosocomial peritonitis (NP) were identified from the 408 admissions occurring during the study period . Patients developing NP were older than the hospitalized CPD patients not developing NP (65.5 +/- 14.6 vs 58.4 +/- 14.7 years, p < 0.05) . Comorbid diseases including diabetes, peripheral vascular disease, gastrointestinal disease, cardiovascular disease, and human immunodeficiency virus seropositivity were not more common in the patients developing NP . Patients developing NP were hospitalized significantly longer than the CPD patients not developing NP (39.5 +/- 46.5 days vs 12.7 +/- 12.4 days, p < 0.001) . The mean serum albumin was lower in the NP patients than in the CPD patients not developing NP (2.35 +/- 0.52 g/dL vs 3.02 +/- 0.60 g/L, p < 0.001) . Antecedent antibiotic use and performance of invasive procedures were noted in 89% and 68% of the patients developing NP, respectively . Staphylococcal species, enterococcal species, and gram-negative organisms accounted for 26%, 21%, and 53% of the episodes of NP, respectively . Furthermore, two strains of Enterococcus resistant to vancomycin were cultured . Eight patients developing NP expired, 8 patients continued CPD therapy, 2 patients transferred to hemodialysis, and one patient recovered renal function . CONCLUSION: We conclude that NP is uncommon . Increased age, increased length of hospital stay, and hypoalbuminemia may predispose patients to the development of NP . Further studies with case controls should help to clarify whether antecedent antibiotics or prior performance of invasive procedures predispose patients to the development of nosocomial peritonitis . The spectrum of organisms accounting for NP is different than the spectrum of organisms causing community-acquired CPD-associated peritonitis . Some of these organisms may be resistant to standard antibiotic therapies . Patients developing NP do poorly, with 42% expiring while being treated for NP. Clin Exp Rheumatol, 1996 Sep-Oct, 14(5), 507 - 12 Experimental septic arthritis in rabbits treated by a combination of antibiotic and steroid drugs; Wysenbeek AJ et al.; OBJECTIVES: Intraarticular steroid injection is traditionally contraindicated during acute septic arthritis . However, there is abundant evidence which proves that the damage to the joint is not only due to the direct effect of bacteria, but also to the local protective mechanisms evoked by the organism . There is, therefore, theoretical justification for a combined therapy of systemic antibiotics and intraarticular corticostertoids in septic arthritis . METHODS: Experimental arthritis was induced by the intraarticular injection of Staphylococcus epidermidis in rabbits . The experimental scheme included three groups of animals: animals that were infected but not treated (group 1); animals treated with systemic antibiotics (group 2); and animals treated with systemic antibiotics and intraarticular steroids (group 3) . Nine days later the animals were sacrificed and joint histopathological-histochemical indices were calculated . RESULTS: Animals from groups 2 and 3 had a smaller pannus, reduced proteoglycan loss, no loss of cartilage height and diminished synovial inflammation in comparison to the animals from group 1 . The animals from groups 2 and 3 were identical in terms of cartilage cellularity, surface erosion, chondrocyte cloning, pannus formation and proteoglycan loss . Synovial inflammation appeared to be less pronounced in group 3 animals when compared to animals of group 2 . CONCLUSION: Concomitant antibiotic-steroid treatment of septic arthritis seems to be harmless in this experimental setting. Optom Vis Sci, 1996 Sep, 73(9), 590 - 4 Factors affecting Staphylococcus epidermidis adhesion to contact lenses; Fleiszig SM et al.; BACKGROUND: Staphylococcus epidermidis is a major causative agent of infectious keratitis associated with contact lens wear . Adhesion of this bacterium to contact lenses may contribute to the pathogenesis of infection and could be influenced by lens surface properties, packaging/storage solutions, and vary among different strains according to the level or type of adhesins expressed . METHODS: Adhesion of six clinical isolates of S . epidermidis to three different contact lens materials was tested . Adhesion assays were performed on lenses immediately after removal from their packages, and also after lenses were soaked in sterile phosphate buffered saline (PBS) for 7 days to dilute the packaging solution . RESULTS: For lenses tested immediately upon removal from their packaging, adhesion to polymacon (in PBS with 0.1% polyvinyl alcohol) was significantly greater than to etafilcon A (in borate buffered saline) and vifilcon A (in PBS) . After soaking, adhesion to polymacon lenses was significantly less than to the other lens materials . This pattern was consistent for all strains, although major differences in baseline adhesion levels existed between strains, with exopolysaccharide (slime)-positive bacteria being more adherent to lenses . CONCLUSIONS: Properties of contact lens materials were not the sole determinant of viable S . epidermidis adhesion to lenses . Strain variability, including levels of exopolysaccharide expression, and the solution used for lens immersion also influenced adhesion. Protein Sci, 1996 Sep, 5(9), 1942 - 6 A fragment of staphylococcal nuclease with an OB-fold structure shows hydrogen-exchange protection factors in the range reported for "molten globules"; Alexandrescu AT et al.; Hydrogen-exchange rates for an OB-fold subdomain fragment of staphylococcal nuclease have been measured at pH 4.7 and 4 degrees C, conditions close to the minimum of acid/base catalyzed exchange . The strongest protection from solvent exchange is observed for residues from a five-stranded beta-barrel in the NMR structure of the protein . Protection factors, calculated from the experimental hydrogen-exchange rates, range between 1 and 190 . Similarly small protection factors have in many cases been attributed to "molten globule" conformations that are supposed to lack a specific tertiary structure . The present results suggest that marginal protection from solvent exchange does not exclude well-defined structure. Protein Sci, 1996 Sep, 5(9), 1898 - 906 A dynamic bundle of four adjacent hydrophobic segments in the denatured state of staphylococcal nuclease; Wang Y et al.; In an earlier study of the denatured state of staphylococcal nuclease (Wang Y, Shortle D, 1995, Biochemistry 34:15895-15905), we reported evidence of a three-strand antiparallel beta sheet that persists at high urea concentrations and is stabilized by a local "non-native" interaction with four large hydrophobic residues . Because the amide proton resonances for all of the involved residues are severely broadened, this unusual structure is not amenable to conventional NMR analysis and must be studied by indirect methods . In this report, we present data that confirm the important role of interactions involving four hydrophobic residues (Leu 36, Leu 37, Leu 38, and Val 39) in stabilizing the structure formed by the chain segments corresponding to beta 1-beta 2-beta 3-h, interactions that are not present in the native state . Glycine substitutions for each of these large hydrophobic residues destabilizes or disrupts this beta structure, as assessed by HN line sharpening and changes in the CD spectrum . The 13C resonances of the carbonyl carbon for several of the residues in this structure indicate conformational dynamics that respond in a complex way to addition of urea or changes in sequence . Studies of hydrogen exchange kinetics in a closely related variant of staphylococcal nuclease demonstrate the absence of the stable hydrogen bonding between the strands expected for a native-like three-strand beta sheet . Instead, the data are more consistent with the three beta strand segments plus the four adjacent hydrophobic residues forming a dynamic, aligned array or bundle held together by hydrophobic interactions. J Hosp Infect, 1996 Sep, 34(1), 31 - 42 A comparison of methods to determine whether clinical isolates of Staphylococcus epidermidis from the same patient are related; Hedin G; Staphylococcus epidermidis is a major cause of hospital-acquired infections but also part of the normal skin flora . A common clinical question is whether repeated isolation of S . epidermidis from one patient represents the same strain; because if different strains are isolated, they are often thought to be contaminants . In this study, different typing methods were compared to answer this question . Twenty isolates of S . epidermidis from five different patients were investigated . The isolates from each patient had identical or very similar antibiograms, and were recovered on different occasions . Typing was performed by antibiogram, biotype, slime production, plasmid profile, and pulsed-field gel electrophoresis (PFGE) banding pattern of SmaI digests of chromosomal DNA . In addition, the level of resistance to methicillin was determined by growth curves in broth containing methicillin for a series of different inocula for each isolate . The results showed that the isolates from each patient belonged to the same clone, but examples of instabilities in their antibiograms, plasmid profiles, as well as their PFGE banding patterns were seen . A change in the level of methicilli, resistance was observed in one strain; otherwise this characteristic was found to be strain-specific and stable in vivo . It was concluded that in combination with biotyping and antibiotic resistance testing the level of resistance to methicillin could be used as an aid to distinguish between two or more clinical isolates of S . epidermidis from the same patient. Int J Food Microbiol, 1996 Sep, 32(1-2), 59 - 71 The combined effects of environmental conditions on lipolysis of pork fat by lipases of the meat starter culture organisms Staphylococcus xylosus and Debaryomyces hansenii; Sorensen BB et al.; The effects of environmental conditions on lipolysis by cell-free extracts from the meat starter culture organisms Staphylococcus xylosus and Debaryomyces hansenii were studied using pork fat emulsions as model systems . For the individual effects of temperature and pH it was found that the optimal conditions for the lipolysis by S . xylosus lipase were 37 degrees C and pH 7.0, and 37 degrees C and pH 6.5 for the lipolysis by D . hansenii lipase . For the combined effects of conditions relevant to meat fermentation, i.e . 10-30 degrees C, pH 4.7-6.0, 2.5-7.5% (w/v) NaCl and incubation times of 2-6 days, the empirical models indicated that temperature, pH and incubation time had important effects on total lipolysis whereas NaCl concentration had little effect . For both cultures lipolysis was strongly inhibited at conditions of meat fermentation compared to optimal conditions . For any set of the conditions which were examined the total lipolysis caused by D . hansenii lipase was lower than that caused by S . xylosus lipase. Antimicrob Agents Chemother, 1996 Sep, 40(9), 2224 - 5 Macrolide-lincosamide-streptogramin B resistance in Staphylococcus lentus results from the integration of part of a transposon into a small plasmid; Werckenthin C et al.; The 8.0-kb macrolide-lincosamide-streptogramin B resistance plasmid pSES20 from Staphylococcus lentus harbored part of a Tn917-like transposon including the left terminal repeat, a gene almost identical to ermB, and its regulatory region, as well as the internal direct repeat . Homology between pSES20 and Tn917 ended at a sequence closely related to those of the resolution sites of Tn917 and Tn552 and staphylococcal recombination sites. Chemotherapy, 1996 Sep-Oct, 42(5), 384 - 90 Antimicrobial prophylaxis with ceftriaxone in neurosurgical procedures . A prospective study of 100 patients undergoing shunt operations; Arnaboldi L; Shunt infection is a major complication of shunt implantation, with Staphylococcus epidermidis found in almost 45% of infected shunts . This pathogen produces an extracellular slime that enables it to adhere to implantable devices and resist antibiotic therapy . Antimicrobial prophylaxis can prevent slime production . In this paper we report the results of a prospective study involving 100 shunt operations . The objective was to evaluate the efficacy of a single preoperative dose of ceftriaxone (2 g, i.v.) in preventing shunt infection . Ceftriaxone was chosen for its pharmacokinetic properties . No shunt infection was observed over a 4-year follow-up period . On the basis of these results we recommend prophylaxis with ceftriaxone as a safe and effective way of preventing shunt infection . In addition, one-shot prophylaxis with ceftriaxone is more cost-effective than multiple-dose antibiotic regimens. Hum Immunol, 1996 Sep 1, 49(2), 113 - 21 Human CD4-reactive antibodies from SLE patients induce reversible inhibition of polyclonal T lymphocyte proliferation; Lenert G et al.; We report on isolation of human polyclonal CD4-reactive antibodies of IgM and/or IgG isotypes from several SLE patients . These antibodies bound specifically to CD4-expressing cell lines and to rCD4 in ELISA and immunoblots . Saturation of CD4-binding sites occurred at antibody concentrations between 5 and 15 micrograms/ml . Anti-CD4 antibodies, in a dose-dependent manner, suppressed the proliferative responses of human peripheral blood mononuclear cells (PBMC) to superantigens (Staphylococcal enterotoxins A and B), anti-CD3 antibodies, and mitogens (PWM and Con A, but not PHA) . They could also inhibit the proliferation of highly purified human T cells induced by immobilized anti-CD3 antibodies . To promote their effects on T cells, human anti-CD4 antibodies had to be present at lymphocyte cultures before or at the time of priming . There was no significant inhibition when antibodies were added more than 24 h following T cell activation . Substantial evidence that the immunosuppression induced by anti-CD4 antibodies was due to their direct effect on T cells was obtained . Down-regulatory effect of anti-CD4 antibodies could be significantly reversed by addition of exogenous IL-2 and by preincubation with soluble recombinant (r)CD4 . Interestingly, at least one affinity-purified anti-CD4 antibody could costimulate the T cell proliferation induced by superantigens or anti-CD3 antibodies, especially when used at subsaturating concentrations (1-4 micrograms/ml) and when added subsequently to the initiation of cultures. J AOAC Int, 1996 Sep-Oct, 79(5), 1095 - 101 Immunoenzymatic detection of staphylococcal enterotoxins: international interlaboratory study; Lapeyre C et al.; An international interlaboratory study was performed by 13 laboratories to validate a commercially available, rapid enzyme immunoassay for detection of staphylococcal enterotoxins A, B, C, D, and E in foods . The 5 enterotoxin serotypes were detected at a level of 0.5 ng/g in mushrooms and ravioli, 0.8 ng/g in meat, 1 ng/mL in milk, and 1.5 ng/g in raw milk cheese when these foods were artificially contaminated with enterotoxin A . Enterotoxins A, B, C, D, or E were also detected in culture supernatants with no protein A interference when normal rabbit serum was used . This method was validated by the French Normalization Agency for the identification of staphylococcal enterotoxins in foods and culture fluids. Eur J Immunol, 1996 Sep, 26(9), 2075 - 80 Synergistic effect between CD40 and class II signals overcome the requirement for class II dimerization in superantigen-induced cytokine gene expression; Mehindate K et al.; Although staphylococcal enterotoxin A (SEA), B (SEB), and toxic shock syndrome toxin 1 (TSST-1) bind to major histocompatibility complex (MHC) class II molecules, they differ in their mode of binding . Signaling induced by these toxins via MHC class II molecules seems to be largely mediated by their mode of interaction . In the present study, we have demonstrated that contrary to SEA, stimulation of the human monocytic cell line THP-1 with SEB or TSST-1 failed to induce interleukin-1 beta or tumor necrosis factor-alpha gene expression . Treatment of THP-1 cells with interferon-gamma increased the level of MHC class II expression but did not enhance the SEB and TSST-1 response . However, cross-linking of SEB or TSST-1 bound to MHC class II molecules with specific antibodies leads to cytokine gene expression, indicating that dimerization of class II molecules is a requirement for this superantigen-induced response . The presence of anti-CD40 antibodies in the course of SEB or TSST-1 stimulation overcomes this requirement, indicating that certain signal(s) induced via CD40 molecules can replace those induced by dimerization of class II molecules . Pretreatment with anti-lymphocyte functional antigen-1 (LFA-1) antibodies completely inhibited SEA-induced response as well as that induced by SEB or TSST-1 in the presence of CD40 antibodies, supporting the involvement of LFA-1 intercellular adhesion molecule system in these responses . The entirety of these results demonstrate clearly that dimerization of class II molecules is a prerequisite for superantigen-induced T cell-independent cytokine gene expression which can be replaced by signaling via CD40 in an LFA-1-dependent system. Arthritis Rheum, 1996 Sep, 39(9), 1499 - 506 Increased responsiveness of rheumatoid factor-producing B cells in seronegative and seropositive rheumatoid arthritis; He X et al.; OBJECTIVE: To compare the frequencies and responsiveness of rheumatoid factor (RF)-producing B cells in the peripheral blood of patients with seronegative and seropositive rheumatoid arthritis (RA) . METHODS: Frequencies of IgM+, IgG+, and RF+ B cells were determined by limiting-dilution analysis of purified peripheral blood B cells from 6 patients with seropositive RA, 8 patients with seronegative RA, and 7 normal controls . B cell help was provided by cloned T helper cells, which were stimulated by either anti-CD3 or the bacterial superantigen staphylococcal enterotoxin D (SED) . IgM and IgG antibodies and RF in culture supernatants were detected by enzyme-linked immunosorbent assay . RESULTS: In the presence of anti-CD3-stimulated T helper cells, 2-10% of B cells from normal individuals secreted IgM and IgG antibodies . The frequency of RF+ B cells was low and ranged from 1:182 to 1:885 (RF+: IgM+) B cells . In patients with seropositive RA, the numbers of Ig-producing B cells were reduced by a factor of 2, while the fraction of RF+ B cell precursors was expanded by more than 50-fold (7-20% of IgM+ B cells; P = 0.004) . Patients with seronegative RA had higher frequencies of RF-producing B cells (1.5-6% of IgM+ B cells) than normal individuals (P = 0.002), but not to the same extent as seropositive patients (P = 0.002) . Stimulation of B cells using SED preferentially induced RF+ B cells in normal controls and in patients with seronegative and seropositive RA . CONCLUSION: B cell precursors with the potential to secrete RF were detectable in high frequencies in normal individuals and in patients with seropositive and seronegative RA . In all donors, these B cells could be stimulated with the bacterial superantigen SED . In normal individuals, RF+ B cells remained nonresponsive to help provided by anti-CD3-activated T cells, but were responsive in RA patients . Seronegative and seropositive RA form a continuous spectrum of disease, with a higher number of RF-secreting B cells in the seropositive patients. Immunity, 1996 Sep, 5(3), 275 - 83 The involvement of an ATP-gated ion channel, P(2X1), in thymocyte apoptosis; Chvatchko Y et al.; In the immune system, apoptosis is involved in intrathymic elimination of self-reactive thymocytes and in peripheral T cell tolerance to exogenous antigens . Here, we describe the role in T cell apoptosis of P(2x1), a nonselective cation channel activated by ATP . P(2X1) molecules are up-regulated in thymocytes during dexamethasone-induced apoptosis, and antagonists to these receptors protect thymocytes from cell death . Moreover, P(2X1) mRNA and protein levels increase in thymocytes induced to die in vivo by the superantigen staphylococcal enterotoxin B . In contrast, T cells undergoing apoptosis in the periphery do not express P(2X1) . The demonstration that P(2X1) ion channels play a role in the apoptosis of thymocytes but not peripheral T cells illustrates a novel mechanism contributing to thymocyte cell death and opens new possibilities for investigating clonal deletion in the thymus. Am J Kidney Dis, 1996 Sep, 28(3), 428 - 36 Risk factors for peritonitis in long-term peritoneal dialysis: the Network 9 peritonitis and catheter survival studies . Academic Subcommittee of the Steering Committee of the Network 9 Peritonitis and Catheter Survival Studies; Golper TA et al.; To determine factors involved in peritoneal dialysis-associated peritonitis and catheter loss, all point prevalent peritoneal dialysis patients in Health Care Finance Administration (HCFA) end-stage renal disease (ESRD) Network 9 were followed throughout 1991 for peritonitis events and throughout 1991 to 1992 for catheter survival . Data were collected by questionnaires compiled by the dialysis facility and validated by network staff . Peritonitis was reported 1,168 times in 729 of the 1,930 patients . By gamma-Poisson regression, a significantly increased risk for peritonitis was observed for patients with previous peritonitis, black race, and those dialyzing with standard connectors or cyclers compared with disconnect systems . Decreased risks were observed for patients with longer ESRD experience and when prophylactic antibiotics were administered before catheter insertion . Postinsertion leakage, diabetes, visual problems, previous or current immunosuppression, and physical activity were not risk factors . Infection of any kind caused the removal of 68% of the 414 catheters lost . Patients with downward-directed tunnels were less likely to experience concomitant exit site/tunnel infections associated with peritonitis . Peritonitis episodes with Staphylococcus epidermidis-like organisms were more likely to resolve with a single course of antibiotics . Perhaps because of their higher infection rate, blacks were more likely than whites to use a disconnect system . In general, the outcome of peritonitis in blacks was similar to that in whites, except that blacks were less likely to be hospitalized and were less likely to die. Scand J Immunol, 1996 Sep, 44(3), 252 - 60 Monocyte-dependent costimulatory effect of TGF- beta 1 on rat T-cell activation; Schiott A et al.; TGF- beta 1 is known to have suppressive effects on both T-cell proliferation and effector functions, but costimulatory effects have also been reported . In the present investigation the effect of TGF- beta 1 is studied in vitro on T-cell proliferative responses of rat spleen cells and of lymph node cells to alloantigens (MLR), the superantigen Staphylococcal enterotoxin A (SEA) or IL-2 . Without addition of TGF- beta 1, adherent, freshly isolated rat spleen monocytes have a suppressive effect on T-cell activation, which upon addition of TGF- beta 1 is reversed to a strong costimulatory effect . The costimulatory effect of TGF- beta 1 is shown to be entirely dependent on the presence of fresh monocytes . Costimulation is demonstrated when TGF- beta 1 is added to spleen cells at the start of the in vitro assays but not when added more than 24 h after the start . Costimulation is not demonstrable when TGF- beta 1 is added to lymph node cells alone but is readily detectable after admixture of freshly isolated spleen monocytes to the lymph node cells . TGF- beta 1 added at the end of culture induces suppression of T-cell activation irrespective of the presence or absence of monocytes . When TGF- beta 1 is added both at the start of an MLC and again after 4 days, the costimulatory effect is maintained, although somewhat moderated . The costimulatory effect of TGF- beta 1 is demonstrated as an increase of the T blast cell population of both CD4+ IL-2R+ and CD8+ IL-2R+ T-cell subsets, whereas the suppressive effect of TGF-beta 1 is shown as reduction of the same parameters. Scand J Immunol, 1996 Sep, 44(3), 229 - 38 Requirement of CD80 and CD86 molecules for antigen presentation by eosinophils; Tamura N et al.; The authors analysed the antigen-presenting ability of eosinophils purified from peritoneal exudate cells of interleukin-5 (IL-5) transgenic mice . The granulocyte-macrophage colony-stimulating factor (GM-CSF)-treated eosinophils induced proliferative responses of primed lymph node T cells and thymus T cells to staphylococcal enterotoxin B (SEB), while untreated eosinophils induced little or no response . GM-CSF-treated eosinophils also induced proliferation of ovalbumin (OVA)-primed lymph node T cells to OVA . Although untreated eosinophils expressed no MHC class II molecule on the surface the eosinophils could be induced to express major histocompatibility complex (MHC) class II molecules when treated with GM-CSF . In the present study, anti-I-Ak monoclonal antibodies (MoAbs) strongly inhibited proliferation of thymus T cells and proliferation of OVA-primed lymph node T cells in response to OVA, but weakly inhibited proliferation of primed T cells in response to SEB . Furthermore, CD80 (B7-1) and CD86 (B7-2) were expressed on the surfaces of untreated eosinophils . The expression of those two molecules on the eosinophils was increased by incubation with GM-CSF . Moreover, anti-CD80 or anti-CD86 MoAbs blocked proliferative responses of primed lymph node T cells and thymus T cells to SEB, and also blocked responses of primed lymph node T cells to OVA . Thus, CD80 and CD86 play an important role in stimulation of T cells by eosinophils. J Immunol, 1996 Sep 1, 157(5), 1935 - 43 Switching on of the proliferation or apoptosis of activated human T lymphocytes by IFN-gamma is correlated with the differential expression of the alpha- and beta-chains of its receptor; Novelli F et al.; To find out how physiologically secreted IFN-gamma controls either the proliferation or the apoptosis of human T lymphocytes, the kinetics of expression of the alpha- and beta-chains of its receptor (IFN-gamma R) were sequentially followed on T lymphocytes first activated with PHA and then cultured in the presence of IL-2, and related to the kinetics of expression of Fas, Bcl-2, and IL-2R p55 chain . Both IFN-gamma R chains were poorly expressed on the membrane of resting T lymphocytes . Following their stimulation with PHA, IFN-gamma R alpha but not IFN gamma R beta-chain up-modulated before T lymphocyte entry into the S phase, and then IFN-gamma R alpha down-modulated when they passed through the S and G2/M . The ensuing proliferative response was inhibited by an anti-IFN-gamma R alpha mAb that impeded the binding of IFN-gamma . When PHA-activated T lymphoblasts were cultured for 16 days with IL-2, IFN-gamma R alpha expression increased, whereas that of the beta-chain remained barely detectable . Fas and Bcl-2 were both highly expressed . When these T lymphoblasts were restimulated by PHA, OKT3, or Staphylococcus enterotoxin beta-pokeweed mitogen, both chains up-modulated and most cells underwent apoptosis in a way apparently independent of Bcl-2, but not of Fas . This apoptosis, too, was prevented by the anti-IFN-gamma R alpha mAb . Physiologically secreted IFN-gamma is thus involved in the activation of resting T lymphocytes and in the apoptosis of reactivated lymphoblasts . However, high expression of IFN-gamma R beta took place when IFN-gamma induced apoptosis, but not when it induced proliferation . In conclusion, a correlation exists between differential expression of the IFN-gamma R beta-chain and the delivery by IFN-gamma of proliferative or apoptotic signals. Infect Immun, 1996 Sep, 64(9), 3893 - 6 The hemagglutinin of Staphylococcus saprophyticus is a major adhesin for uroepithelial cells; Meyer HG et al.; The 160-kDa hemagglutinin of Staphylococcus saprophyticus also serves as a fibronectin-binding protein, and the two activities may be present on different parts of the molecule . Bacteria expressing the 160-kDa hemagglutinin bound in large numbers to histological sections of human ureters, whereas nonhemagglutinating bacteria did not bind . Binding was decreased by an antiserum to the 160-kDa protein and by a preparation of sheep erythrocyte membranes . Fibronectin had no effect . We therefore conclude that binding of S . saprophyticus to uroepithelial cells is mediated by the hemagglutinating activity of the 160-kDa surface protein. J Urol, 1996 Sep, 156(3), 991 - 4 A randomized prospective comparison of antibiotic tissue levels in the corpora cavernosa of patients undergoing penile prosthesis implantation using gentamicin plus cefazolin versus an oral fluoroquinolone for prophylaxis; Schwartz BF et al.; PURPOSE: We performed a prospective randomized trial comparing the efficacy, safety and cost of parenteral antibiotics and oral fluoroquinolones for prophylaxis in penile prosthesis surgery . MATERIALS AND METHODS: We prospectively randomized 20 consecutive patients undergoing penile prosthesis surgery to receive ofloxacin orally or gentamicin and cefazolin parenterally followed by cephradine orally . Intraoperatively corpora cavernosa tissue and simultaneous peripheral serum samples were evaluated for antibiotic levels . Median followup was 16 months (range 8 to 21, mean 15.35) . RESULTS: There were no implant losses or reoperations and complications were comparable in the 2 groups . The difference in mean serum-to-tissue ratios of ofloxacin versus the combination of cefazolin and gentamicin was statistically significant (p < 0.03) . The minimum inhibitory concentrations of ofloxacin met or exceeded those of the 2 most common offending organisms, Staphylococcus epidermidis and Escherichia coli, in 80% of patients, which was comparable to the results of the parenteral regimen . Cost savings of the medications alone were more than $250,000 in the ofloxacin group . By eliminating a hospital stay of the 25,000 cases of penile prosthesis placement in the United States yearly a total cost savings of more than $36 million would be realized . CONCLUSIONS: When oral ofloxacin is given for prophylaxis in penile implant surgery, the procedure may be performed on an outpatient basis and health care dollars are saved. J Biol Chem, 1996 Aug 30, 271(35), 21075 - 80 Limited proteolysis of rat phosphatidylinositol transfer protein by trypsin cleaves the C terminus, enhances binding to lipid vesicles, and reduces phospholipid transfer activity; Tremblay JM et al.; Rat phosphatidylinositol transfer protein (PITP) is a 32-kDa protein of 271 amino acids that transfers phosphatidylinositol and phosphatidylcholine between membranes . The alpha isoform of rat PITP was expressed in Escherichia coli and purified in high yields . The purified protein contained 1 mol of phosphatidylglycerol and had a transfer activity for phosphatidylinositol and phosphatidylcholine equal to or greater than that of PITP purified from mammalian brain . Limited protease digestion was used to further define structure, activity, and function relationships in PITP . PITP alone is relatively resistant to digestion by chymotrypsin, trypsin, and Staphylococcus V8 protease but is readily cleaved by subtilisin . Phospholipid vesicles containing phosphatidic acid enhance susceptibility to digestion by all four proteases . In the presence of vesicles, PITP, which migrates as a 36-kDa protein in SDS-polyacrylamide gel electrophoresis, is cleaved rapidly by trypsin to a form that appears to be 2-3 kDa smaller than the native form . The tryptic fragment retains partial phospholipid transfer activity and shows an enhanced affinity for phospholipid vesicles containing phosphatidic acid . Analysis of the tryptic digestion products by immunoblotting, N-terminal sequencing, and electrospray mass spectrometry showed that trypsin cleaves the C terminus of PITP at Arg253 and Arg259 . Thus, removal of the C terminus enhances the affinity of PITP for vesicles and results in a dimunition of transfer activity . Overall, the data show that PITP undergoes conformation changes and that the C terminus becomes more accessible to trypsin when bound to vesicles . Hence, the C terminus is not an essential component of the membrane binding site and may be located distal to it. Cell Immunol, 1996 Aug 25, 172(1), 135 - 8 Anti-HLA class I antibody inhibits Staphylococcus enterotoxin A (SEA)-induced proliferation of human PBMC; Baskar P et al.; The antigen-presenting role of major histocompatibility complex (MHC) Class I molecules in the activation of appropriately restricted T cells is well documented . Now growing evidence indicates that MHC Class I molecules can, in addition, exert a regulatory effect and influence the resulting immune responses . In this report we show that a monoclonal antibody (mAb) directed against a conserved region of the human leukocyte antigens (HLA)-A, -B, and -C was able to inhibit proliferation of human peripheral blood mononuclear cells induced by the superantigen, Staphylococcus enterotoxin A . While anti-HLA inhibition was associated with a decrease in IL-2 receptor (IL-2R) expression, the addition of exogenous IL-2 did not restore the proliferative response in the presence of anti-HLA mAb . The inhibition of DNA synthesis was also associated with a decrease in the expression of the early activation marker CD69 . These results suggest a critical role for HLA Class I molecules in the early events of human lymphocyte activation and proliferation as well as in their expression of the IL-2R. Vet Rec, 1996 Aug 24, 139(8), 185 - 7 Tylosin in the treatment of canine superficial pyoderma; Harvey RG; Thirty dogs with superficial pyoderma were treated orally with tylosin at a dose of 20 mg/kg twice daily for three weeks . Staphylococcus intermedius was recovered from 21 (70 per cent) of the dogs . Twenty-two of the dogs were free of clinical signs after three weeks of treatment and two dogs responded to a further two weeks of treatment, giving a total response rate of 80 per cent . Five cases (16.6 per cent) failed to respond and three of these subsequently responded to other antibacterial treatment . One dog suffered transient gastritis after the doses of tylosin and subsequently responded to a different antibacterial agent. J Mol Biol, 1996 Aug 23, 261(3), 372 - 85 Capsid targeting sequence targets foreign proteins into bacteriophage T4 and permits proteolytic processing; Mullaney JM et al.; A membrane-independent morphogenetic viral signal peptide is identified within bacteriophage T4 internal protein III (IPIII) . Utilizing a phagederived expression-packaging-processing system, which packages foreign proteins fused with IPIII into the phage capsid, a synthetic cleavage site introduced at the C terminus of IPIII, is demonstrated to be functional and permits processing of fusion proteins . IPIII, which possesses a native P21 cleavage site at its N terminus, is altered to possess a second P21 cleavage site at its C terminus where cleavage occurs by means of the scaffold proteinase P21 within the capsid . The altered IPIII was inserted into an expression vector to permit the creation of fusion proteins with staphylococcal nuclease, EcoRI endonuclease, beta-globin, and luciferase . Western immunoblot analysis of packaged T4eG326 indicates that the IPIII:fusion-proteins are packaged into phage and proteolytically processed, thus the synthetic P21 cleavage site positioned at the C terminus of IPIII is demonstrated to be functional, and 20 to 200 protein molecules are packaged per capsid . Truncation experiments identified the minimal portion of IPIII required to achieve targeting into the phage capsid as a ten amino acid residue from the N terminus, which includes the N-terminal methionine residue and the proteinase P21 cleavage site, designated the CTS (capsid targeting sequence) . The addition of the CTS to a fragment of luciferase permits the protein to be packaged and processed, which demonstrates that the CTS is by itself sufficient to target foreign protein to the capsid . The imputed dual function of the CTS is supported by site-directed PCR mutagenesis, which reveals two functionally separate domains of the CTS for targeting and processing . The CTS appears to function in a core-related targeting mechanism that directs a polymorphic set of proteins into the T-even capsid or scaffold . Although structure formation is often assumed to involve extended protein interfaces, the analysis shows that a limited but specific sequence, the CTS, drives the interaction required to achieve targeting. Eur J Biochem, 1996 Aug 15, 240(1), 215 - 22 The use of photolabelled peptides to localize the substance-P-binding site in the human neurokinin-1 tachykinin receptor; Girault S et al.; The amino acid p-benzoyl-L-phenylalanine, (p-Bz)Phe, has been incorporated into substance P (SP), Arg-Pro-Lys-Pro-Gln-Gln-Phe-Phe-Gly-Leu-Met-NH2, to localize the agonist-binding domains of the human neurokinin-1 (NK-1) receptor overexpressed in a transfected mammalian cell line . The NK-1-specific agonist {Pro9}SP was modified at position 8 by (p-Bz)Phe and acylated at the N-terminus by a biotinyl sulfone reporter via a 5-aminopentanoyl spacer . After photolysis, the biotinyl sulfone moiety allowed easy and efficient removal of biotinylated fragments from the complex incubation mixture with streptavidin-coated beads . Direct elution from the beads with the matrix used for matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOFMS), which was facilitated by saturation of streptavidin sites with biotin, and subsequent MALDI-TOF mass spectrometry analysis allowed identification of the NK-1 fragments obtained after photolysis and proteolytic digestion . Trypsin digestion and combined trypsin/Staphylococcus aureus V8 protease enzymatic cleavage established that the site of covalent attachment of the photolabelled SP resides in the second extracellular loop Thr173-Arg177 . Cyanogen bromide cleavage shows that the probe is covalently attached to the methyl group of a methionine residue from human NK-1 . These experiments identified Met174 as the modified residue. FEMS Microbiol Lett, 1996 Aug 15, 142(1), 59 - 63 Attachment of bacteria to model solid surfaces: oligo(ethylene glycol) surfaces inhibit bacterial attachment; Ista LK et al.; Bacterial cell attachment to the surfaces of self-assembled monolayers formed by the adsorption of omega-substituted alkanethiols on transparent gold films has been studied under defined bacterial culture and flow conditions . Phase contrast microscopy was used to quantify the attachment of two organisms, one of medical (Staphylococcus epidermidis) and one of marine (Deleya marina) importance . Self-assembled monolayers terminated with hexa(ethylene glycol), methyl, carboxylic acid and fluorocarbon groups were investigated . Over the range of experimental conditions, self-assembled monolayers formed from HS(CH2)11(OCH2CH2)6OH were found to be uniformly resistant to bacterial attachment, with a 99.7% reduction of attachment for both organisms when compared to the most fouled surface for each organism . On other surfaces, S . epidermidis and D . marina were shown to exhibit very different attachment responses to the wettability of the substratum . While the attachment of S . epidermidis correlated positively with surface hydrophilicity, D . marina showed a preference for hydrophobic surfaces . This study suggests that surfaces incorporating high densities of oligo(ethylene glycol) are good candidates for surfaces that interact minimally with bacteria. J Immunol, 1996 Aug 15, 157(4), 1758 - 72 Inhibition of superantigen-induced proinflammatory cytokine production and inflammatory arthritis in MRL-lpr/lpr mice by a transcriptional inhibitor of TNF-alpha; Edwards CK 3rd et al.; We have used fas-defective MRL-lpr/lpr mice to study the effects of the staphylococcal enterotoxin superantigens on the development of autoimmune, inflammatory joint disease in animals that are susceptible to the development of rheumatoid arthritis-like disease . We show that systematic administration by a single i.p . injection of staphylococcal enterotoxin B (SEB; 10 micrograms/mouse) caused a mild, inflammatory arthritis +30 days postchallenge in the knee joints of young (< 2-mo-old) MRL-lpr/lpr mice, but not aged-matched MRL +/+ mice . In aged (> 8-mo-old) MRL-lpr/lpr mice, but not in aged MRL +/+ mice, SEB caused a severe, inflammatory arthritis, as assessed histologically, and systemic autoimmune disease, including glomerulonephritis and autoantibody production . Furthermore, in aged MRL-lpr/lpr mice, SEB but not heat-denatured SEB caused acute weight loss and elevated levels of serum proinflammatory cytokines . Compared with highly purified peritoneal macrophages obtained from either aged MRL +/+, young MRL-lpr/lpr, or young MRL +/+, peritoneal macrophages obtained from aged MRL-lpr/lpr mice constitutively expressed 2- to 10-fold greater levels of TNF-alpha, IL-1 beta, IL-6, and IL-10, and produced elevated amounts of these cytokines when treated in vitro with SEB . SEB-challenged aged MRL-lpr/lpr mice treated with anti-TNF mAb (100 micrograms/mouse; every other day), anti-V beta 8 TCR mAb (250 micrograms/mouse; every other day), or orally with the novel TNF-alpha inhibitor MDL 201,449A (9-{(1R, 3R)-trans-cyclopentan-3-ol} adenine; 25 mg/kg/day) exhibited reduced inflammatory arthritis, autoantibody formation, and serum TNF-alpha levels, but not IL-10 levels, after +30 days of treatment . These data suggest that SEB is an extremely potent macrophage-activating factor in vitro and in vivo, enhancing several aspects of autoimmune disease in MRL-lpr/lpr mice, and that anti-TNF therapies may have potential use in inflammatory arthritis. J Immunol, 1996 Aug 15, 157(4), 1422 - 31 Implantation of IL-2-containing osmotic pump prolongs the survival of superantigen-reactive T cells expanded in mice injected with bacterial superantigen; Kuroda K et al.; In the present study we investigated the mechanism of deletion of superantigen (sAg)-reactive T cells expanded in sAg-injected mice . In staphylococcal enterotoxin A (SEA)-injected mice, IL-2 activity in serum peaked at 1 to 3 h and the expression of IL-2R alpha-chain (IL-2R alpha) on SEA-reactive (V beta 3+, or V beta 11+) T cells peaked at 6 to 12 h after the injection . Expansion of V beta 3+ or V beta 11+ T cells peaked at 2 days after the injection when most of these T cells were IL-2R alpha negative, and IL-2 activity was not detected at all in serum, suggesting the involvement of IL-2 deprivation in the deletion of expanded T cells . Implantation of an osmotic pump containing human rIL-2 (IL-2 pump) prolonged the expanded states of V beta 3+ or V beta 11+ T cells in SEA-injected C57BL/6 mice and of V beta 8+ T cells in SEB-injected MRL +/+ and Fas Ag-defective MRL-Ipr/Ipr mice . Adult thymectomy did not change at all the effect induced by implantation of IL-2 pump . DNA fragmentation was blocked substantially in mice co-treated with SEA and IL-2 pump . In addition, CD4+ T cell blasts, obtained by in vitro stimulation with rIL-2 of splenic CD4+ T cells from mice co-treated with SEA and IL-2 pump, produced substantial amounts of IL-2 upon restimulation with SEA . These results indicate that deprivation of IL-2 is deeply involved in the deletion of expanded sAg-reactive T cells and their anergy induction in sAg-injected mice. Cancer Res, 1996 Aug 15, 56(16), 3731 - 6 Augmentation of antitumor immunity with bacterial superantigen, staphylococcal enterotoxin B-bound tumor cells; Shimizu M et al.; We have examined the effectiveness of a bacterial superantigen, staphylococcal enterotoxin B (SEB)-coupled tumor cells, to induce antitumor activity . SEB was chemically conjugated to tumor cells using a heterobifunctional cross-linking agent acting through NH2 and SH groups . V beta 8+ T cells were activated and increased in number after the culture with SEB-bound Meth A cells . The cultured T cells exhibited an antitumor activity in the Winn assay . Interleukin 2 (IL-2) receptor alpha + (IL-2R+) V beta 8+ T cells but not IL-2R+ V beta 6+ T cells increased in number in mice injected with SEB-bound Meth A cells . However, the percentages of V beta 8+ and V beta 6+ T cells did not change by this immunization . The antitumor effector cells were V beta 7- 8- CD4+ T cells . In vivo immunization with SEB-bound cells induced a strong antitumor activity, i.e., tumor-free mice/total mice = 14 of 15 (93%) for Meth A and 7 of 15 (47%) for hepatoma MH134 . The induced antitumor activity was both dose dependent and tumor specific . Treatment with SEB-bound cells prolonged the survival days of Meth A-bearing mice by 62% . These results suggest that SEB-bound tumor cells may be a powerful method for induction of in vivo antitumor activity. Biochemistry, 1996 Aug 13, 35(32), 10441 - 7 Identification from a phage display library of peptides that bind to toxic shock syndrome toxin-1 and that inhibit its binding to major histocompatibility complex (MHC) class II molecules; Sato A et al.; Phage display technique is a powerful tool with which to identify novel binding sequences for antibody and receptor targets . Few studies, however, have used this technology to select affinity peptides for ligand molecules . Here, we screened a peptide phage library for binding to toxic shock syndrome toxin 1 (TSST-1) to examine whether peptide ligands for TSST-1 which mimic the structure of major histocompatibility complex (MHC) class II receptors could be identified . After three cycles of biopanning, four potent sequences reactive with TSST-1 were isolated (designated phages 2, 3, 8, and 11) . Selected phage were found to react specifically with TSST-1 but not with other staphylococcal exotoxins . A synthetic peptide (pep3) corresponding to the most frequently identified sequence (phage3) was shown to inhibit binding of all four isolated phage to TSST-1, suggesting that they bind to a common site on TSST-1 . Furthermore, pep3 was shown to compete with MHC class II molecules for binding to TSST-1 in a concentration-dependent manner . Comparison of their sequences with MHC class II molecules revealed that phage8 shared sequence homology with two regions of the beta chain of MHC class II molecules: amino acids 57-62, containing a residue (Tyr-60) involved in TSST-1 binding as suggested by X-ray crystallographic data of TSST-1-MHC class II complex; and amino acids 188-193, a region not previously known as a contact domain . These results suggest that the selected sequences recognized the MHC class II binding site on TSST-1 . Thus, affinity selection for peptides binding to ligand molecules (e.g., TSST-1) rather than their cognate receptors (e.g., MHC class II) from a random phage display library represents a useful approach to understanding receptor-ligand interactions. Biochemistry, 1996 Aug 13, 35(32), 10328 - 38 Engineered disulfide bonds in staphylococcal nuclease: effects on the stability and conformation of the folded protein; Hinck AP et al.; Efforts to enhance the stability of proteins by introducing engineered disulfide bonds have resulted in mixed success . Most approaches to the prediction of the energetic consequences of disulfide bond formation in proteins have considered only the destabilizing effects of cross-links on the unfolded state (chain entropy model) {Pace, C . N., Grimsley, G . R., Thomson, J . A., & Barnett, B . J . (1988) J . Biol . Chem . 263, 11820-11825: Doig, A . J., & Williams, D . H . (1991) J . Mol . Biol . 217, 389-398} . It seems clear, however, that disulfide bridges also can influence the stability of the native state . In order to assess the importance of the latter effect, we have studied four variants of staphylococcal nuclease (V8 strain) each containing one potential disulfide bridge created by changing two wild-type residues to cysteines by site-directed mutagenesis . In each case, one of the introduced cysteines was within the type VIa beta turn containing cis Pro117, and the other was located in the adjacent extended loop containing Gly79 . In all four cases, the overall loop size was kept nearly constant (the number of residues in the loop between the two cysteines varied from 37 to 42) so as to minimize differences from chain entropy effects . The objective was to create variants in which a change in the reduction state of the disulfide would be coupled to a change in the position of the equilibrium between the cis and trans forms of the Xxx116-Pro117 peptide bond in the folded state of the protein . The position of this equilibrium, which can be detected by NMR spectroscopy, has been shown previously to correlate with the stability of the native protein . Its determination provides a measure of strain in the folded state . The thermal stabilities and free energies for unfolding by elevated temperature and guanidinium chloride were measured for each of the four mutants under conditions in which the introduced cysteines were cross-linked (oxidized) and unlinked (reduced) . In addition, reduction potentials were determined for each mutant . Formation of the different disulfide bridges was found to induce varying levels of folded state strain . The stabilization energy of a given disulfide bridge could be predicted from the measured perturbation energy for the peptide bond isomerization, provided that energetic effects on the unfolded state were calculated according to the chain entropy model . Undiagnosed strain in native states of proteins may explain the variability observed in the stabilization provided by engineered disulfide bridges. Biochemistry, 1996 Aug 6, 35(31), 10234 - 9 Testing the correlation between delta A and delta V of protein unfolding using m value mutants of staphylococcal nuclease; Frye KJ et al.; The application of hydrostatic pressure to aqueous protein solutions results in the unfolding of the protein structure because the protein-solvent system volume is smaller for the unfolded state . Contributions to this decrease in volume upon unfolding (delta Vu) derive from altered interactions of the protein with solvent and are presumed to include electrostriction of charged residues, elimination of packing defects, and hydration of hydrophobic surfaces upon unfolding . If the contribution of hydrophobic surface area solvation to the observed volume change of unfolding were large and negative, as is generally assumed, then one would expect to find a correlation between the amount of surface area exposed on unfolding, delta A(u), and the volume change, delta Vu . In order to test this correlation, we have determined delta Vu for two mutants of staphylococcal nuclease, A69T + A90S and H121P, whose unfolding by denaturant is, respectively, either significantly more (28%) or significantly less (28%) cooperative than that observed for wild-type (WT) . This cooperativity coefficient or m value has been shown to correlate with delta A(u) . If, in turn, delta Vu is correlated with delta A(u), we would expect the m+ mutant, A69T + A90S, to exhibit a delta Vu that is more negative than WT nuclease, while the delta Vu for the m- mutant, H121P, should be smaller in absolute value . To verify the correlation between m value and delta A(u) for these mutants, we determined the xylose concentration dependence of the stability of each mutant at atmospheric pressure and as a function of pressure . The efficiency of xylose stabilization was found to be much greater for the m+ mutant than for WT, consistent with an increase in delta A(u), while that of the m- mutant was found to be only slightly greater than for WT, indicating that other factors may contribute to the denaturant m value in this case . Regardless of the denaturant m value or the effect of xylose on stability, the volume changes upon unfolding for both mutants were found to be within error of that observed for WT . Thus, there does not appear to be a correlation between the volume change and the change in exposed surface area upon unfolding . We have previously shown a lack of pH dependence of the volume change, ruling out electrostriction as a dominant contribution to delta Vu of nuclease . These studies implicate either compensation between polar and nonpolar hydration or excluded volume effects as the major determinant for the value of delta Vu. Exp Eye Res, 1996 Aug, 63(2), 129 - 36 Aqueous and vitreous penetration of ciprofloxacin following different modes of systemic administration; Madu AA et al.; The overall importance of the peak or the mean serum concentrations as predictors of ocular drug penetration is unknown . To address this fundamental question with an agent which shows promise as adjunctive therapy in the treatment of endophthalmitis, we studied the penetration of ciprofloxacin into the aqueous and vitreous humors following three different modes of systemic administration . New Zealand white rabbits received either a single bolus dose (40 mg kg-1), three intermittent doses of 13.33 mg kg-1 evenly spaced over an 8 hr period, or a continuous infusion of 40 mg kg-1 over an 8 hr period . Pharmacokinetic analysis was performed using RSTRIP II, a non-linear, least square regression model analysis program . The serum area under the concentration-time curve (AUC) values for each mode of drug administration were similar: 32.9 micrograms hr ml-1 for single dose, 31.9 micrograms hr ml-1 for intermittent dose, and 33.8 micrograms hr ml-1 for continuous infusion modes . The percentage penetration into the aqueous and vitreous were also similar; 30.5% and 6.5% for a single dose, 31.6% and 7.4% for intermittent doses and 30.0% and 7.5% for continuous infusion . The penetration into the aqueous and vitreous humors was not influenced by mode of administration . As with other quinolones we have studied, elimination rates were similar for the central and peripheral compartments in the post-distributive phase . Vitreous humor ciprofloxacin concentrations achieved were below that which inhibits most Staphylococcus epidermidis, the most common isolate in patients with post-operative endophthalmitis. Vet Immunol Immunopathol, 1996 Aug, 52(4), 301 - 11 Analysis of monoclonal antibodies reactive with molecules upregulated or expressed only on activated lymphocytes; Davis WC et al.; Monoclonal antibodies potentially specific for antigens expressed or upregulated on activated leukocytes were selected for further analysis from the panel submitted to the third international workshop on ruminant leukocyte antigens . The kinetics of expression of these activation antigens on resting peripheral mononuclear cells (PBMC) and PBMC stimulated with concanavalin A or staphylococcal superantigen SECI for 4, 24 or 96 h were compared, as well as their appearance on various subsets of cells . For some of them, a molecular mass could be determined after immunoprecipitation from radio-labeled, lectin-stimulated cells . Based on the results from the clustering, kinetic studies and biochemical data, evidence was gathered for assigning two additional mAbs to cluster BoCD25 (IL-2 receptor) and two mAbs to cluster BoCD71 (transferrin receptor) . Four mAbs recognized an early activation antigen predominantly expressed on gamma delta T cells in short-term cultures . A number of other activation antigens were further characterized. Immunology, 1996 Aug, 88(4), 611 - 7 Isotype and antibody specificity of spontaneously formed immunoglobulins in pig fetuses and germ-free piglets: production by CD5- B cells; Cukrowska B et al.; Pig fetuses, colostrum-deprived newborns and germ-free (GF) piglets, animals in which B-cell development is not influenced by maternal regulatory factors, were employed to study the occurrence and specificity of natural antibodies (NAb) . Serum immunoglobulins of all isotypes were found in 44-day-old fetuses (the gestation period in pigs lasts 114 days) and their level, with predominating IgM, was increased during fetal ontogeny . In sera of fetuses at the end of embryonic life as well as of newborns and older GF piglets, antibody activity against autoantigens (thyroglobulin, hormones, ssDNA), phylogenetically conserved proteins (myosin), haptens (trinitrophenyl; TNP) and bacterial components (Escherichia coli O86, tetanic anatoxin) was detected by enzyme-linked immunosorbent assay . The antigen-biding activity of IgM NAb increased after isolation of the serum immunoglobulins on a Staphylococcus Protein A (SPA)-Sepharose column . IgM reactivity similar to that detected in serum was found in supernatants from polyclonally stimulated cultures of spleen of 8- and 12-day-old GF piglets . Pig fetal liver IgM+ B cells, which were able to produce IgM after polyclonal stimulation, did not express the CD5 molecule . Our results indicate that pig preimmune repertoire is comparable to that described in humans and mice, although in contrast to these species pig B-1 cells do not express CD5. Immunology, 1996 Aug, 88(4), 586 - 92 Activation by mitogens and superantigens of axolotl lymphocytes: functional characterization and ontogenic study; Salvadori F et al.; Urodele amphibians have weak and slow immune responses compared to mammals and anuran amphibians . Using new culture conditions, we tested the ability of lymphocytes of a well-studied salamander, the Mexican axolotl (Ambystoma mexicanum) to proliferate in vitro with diverse mitogenic agents . We demonstrated that the axolotl has a population of B lymphocytes that proliferate specifically and with a high stimulation index to the lipopolysaccharide (LPS) known as a B-cell mitogen in mammals . This proliferative capacity is observed without significant changes throughout ontogenesis . In the presence of LPS, axolotl B lymphocytes are able to synthesize and secrete both isotopes of immunoglobulin described in this species, IgM and IgY . Moreover, a distinct lymphocyte subpopulation is able to poliferate significantly in response to the mitogens usually known as T-cell specific in mammals, phytohaemagglutinin (PHA) and concanavalin A (Con A) . The activated cells are T lymphocytes, as shown by depletion experiments performed in vitro with monoclonal antibodies, and in vivo by thymectomy . Splenic T lymphocytes of young axolotls (before 10 months) do not have this functional ability, which suggests maturation and/or migration phenomena during T-cell ontogenesis in this species . Axolotl lymphocytes are able to proliferate in vitro with a significant stimulation index to staphylococcal enterotoxins A and B (SEA and SEB) . These products act on mammalian lymphocytes as superantigens: in combination with products of the major histocompatibility complex (MHC), they bind T-cell receptors with particular V beta elements . The fact that these superantigens are able to activate lymphocytes of a primitive vertebrate suggests a striking conservation of molecular structures implied in superantigen presentation and recognition. Alcohol Clin Exp Res, 1996 Aug, 20(5), 900 - 7 Regulation of human monocyte functions by acute ethanol treatment: decreased tumor necrosis factor-alpha, interleukin-1 beta and elevated interleukin-10, and transforming growth factor-beta production; Szabo G et al.; We and others have previously shown that even acute ethanol exposure has the capacity to modulate immune functions, particularly monocyte functions . Herein, we tested the hypothesis that acute ethanol treatment inhibits inflammatory, while increasing inhibitory cytokine production in human blood monocytes that, in turn, could contribute to the overall immune abnormalities seen after alcohol use . Our data show that in vitro treatment of blood monocytes with a physiologically relevant dose of alcohol (25 mM) results in significantly decreased induction of tumor necrosis factor-alpha (TNF alpha) and interleukin (IL)-1 beta by bacterial stimulation of either Gram-positive {staphylococcal enterotoxin B (SEB), 1 microgram/ml of SEB} or Gram-negative {lipopolysaccharide (LPS), 1 microgram/ml of LPS} origin both at the protein and mRNA levels . In contrast, acute ethanol treatment induces monocyte production of mediators with immunoinhibitory potential, including transforming growth factor-beta and IL-10 . We further show that ethanol not only induces monocyte/macrophage (Mo) IL-10 and transforming growth factor-beta, but even augments bacterial (both LPS and SEB) stimulation-induced production of both of these cytokines . IL-10 is a potent inhibitor of Mo TNF alpha production . We found that ethanol-induced elevation in Mo IL-10 levels contributes to the decreased Mo TNF alpha production to bacterial challenge in ethanol-exposed Mo . However, mRNA levels for TNF alpha are downregulated as early as 1.5 hr after ethanol treatment, suggesting that ethanol likely has an IL-10 independent, direct effect on early signaling events of TNF alpha induction. J Dairy Res, 1996 Aug, 63(3), 361 - 8 Field study on the relationship between teat thickness changes and intramammary infections; Zecconi A et al.; The aim of this study was to describe the results of teat thickness measurement applied routinely in three commercial dairy herds and to evaluate the influence of machine-induced teat thickness changes on intramammary infection and the frequency of new infection . A total of 1018 fore milk samples and the same number of teat apex measurements have been evaluated . Overall, relative teat thickness changes were normally distributed (mean -0.16%, SD 10.15%), while a specific pattern could be observed within herds . Increases in teat thickness of > 5% were significantly associated with infection and new infection (odds ratio > 1), but the association was not significant when teat thickness decreased by more than 5% . When results were classified according to aetiology, analysis showed that coagulase-negative staphylococcal infections were significantly associated with both increases and decreases in teat thickness numerically greater than 5%. Spine, 1996 Aug 1, 21(15), 1820 - 3 Conservative therapy of atlantoaxial osteomyelitis . A case report; Lam CH et al.; STUDY DESIGN: A rare case of C1-C2 vertebral osteomyelitis treated conservatively is described . The radiologic findings as well as the follow-up evaluation are reported . OBJECTIVE: To increase knowledge about the pathogenesis and treatment of vertebral osteomyelitis in the high cervical region . SUMMARY OF BACKGROUND DATA: This is one of the first cases reported of successful conservative treatment of osteomyelitis at this level . METHODS: In a 58-year-old man with lumbar staphylococcal infection, a subsequent cervical infection developed . Because the lumbar spondylitis was treated promptly, the cervical osteomyelitis was treated at a very early stage of development . RESULTS: Operative decompression is the treatment most often used in osteomyelitis at the C1-C2 level . This is an extremely unusual circumstance in which early treatment of the infection negated the need for surgery . CONCLUSION: Conservative treatment of osteomyelitis at the C1-C2 level can be efficacious in the correct setting. Protein Sci, 1996 Aug, 5(8), 1737 - 41 Crystal structure of a biologically inactive mutant of toxic shock syndrome toxin-1 at 2.5 A resolution; Papageorgiou AC et al.; Toxic shock syndrome toxin-1 (TSST-1) is one of a family of staphylococcal exotoxins recognized as microbial superantigens . The toxin plays a dominant role in the genesis of toxic shock in humans through a massive activation of the immune system . This potentially lethal illness occurs as a result of the interaction of TSST-1 with a significant proportion of the T-cell repertoire . TSST-1, like other superantigens, can bind directly to class II major histocompatibility (MHC class II) molecules prior to its interaction with entire families of V beta chains of the T-cell receptor (TCR) . The three-dimensional structure of a mutant (His-135-Ala) TSST-1 was compared with the structure of the native (wild-type) TSST-1 at 2.5 A resolution . The replacement of His 135 of TSST-1 with an Ala residue results in the loss of T-cell mitogenicity and toxicity in experimental animals . This residue, postulated to be directly involved in the toxin-TCR interactions, is located on the major helix alpha 2, which forms the backbone of the molecule and is exposed to the solvent . In the molecular structure of the mutant toxin, the helix alpha 2 remains unaltered, but the His to Ala modification causes perturbations on the neighboring helix alpha 1 by disrupting helix-helix interactions . Thus, the effects on TCR binding of the His 135 residue could actually be mediated, wholly or in part, by the alpha 1 helix. Nippon Kyobu Geka Gakkai Zasshi, 1996 Aug, 44(8), 1185 - 92 {A case report of active aortic prosthetic valve endocarditis due to methicillin-resistant Staphylococcus epidermidis}; Asakura T et al.; A 71-year-old woman with active aortic prosthetic endocarditis due to Methicillin-resistant Staphylococcus epidermidis (MRSE) and subannular mycotic aneurysm and paravalvular leakage and acute mitral regurgitation underwent emergent surgical treatment . The mycotic aneurysm was closed using a prosthetic patch after surgical debridement . Re-aortic valve replacement with a 21-mm Hancock II prosthesis was performed at the paraannular position by utilizing the patch . Mitral valve was also replaced with a 27-mm Hancock II prosthesis . Antibiotic therapy was provided by vancomycin combined with rifampicin and gentamicin . The following regimen was given, vancomycin 1 g i.v . q12h for 6 weeks plus gentamicin 80 mg/day i.v . for 4 weeks plus rifampicin 450 mg/day orally for 6 weeks . Vancomycin and gentamicin doses were modified appropriately according to the monitored serum levels in the patient with renal failure . Postoperative course was uneventful . The patient is doing well 11 months after surgery and no recurrence of infection has been seen . We conclude that prompt surgical removal of the infected sources and appropriate antibiotic therapy based on the bacteriology may be the only curative treatment for uncontrolled infection at the active phase of MRSE prosthetic endocarditis. Cell Immunol, 1996 Aug 1, 171(2), 240 - 5 Superantigen-induced Langerhans cell depletion is mediated by epidermal cell-derived IL-1 alpha and TNF alpha; Shankar G et al.; The purpose of this study was to examine the role of cytokines in staphylococcal enterotoxin-A (SEA)-induced epidermal Langerhans cell (LS) depletion . This was accomplished by analyzing the effect of SEA on cytokine secretion by cultured epidermal cell populations by means of ELISA and by assessing the capacity of cytokines and cytokine-specific antibodies to affect epidermal LC density (as measured by immunoperoxidase staining of epidermal cells bearing surface Ia) . The results of this study indicate that SEA induces the secretion of IL-1 alpha and TNF alpha by epidermal cells, that these cytokines induce LC depletion from epidermis, and that antibodies specific for these agents inhibit the depletion of LC by SEA . Taken together, these findings suggest that IL-1 alpha and TNF alpha may play essential roles in SEA-mediated depletion of LC from murine epidermis. J Pharmacol Exp Ther, 1996 Aug, 278(2), 800 - 7 Agonist synergism in airway smooth muscle contraction; Gerthoffer WT; Hyperreactive responses of asthmatics to bronchoconstrictor agonists are caused by mediators produced by a variety of cell types within the airways . Nerves, mast cells, eosinophils, airway epithelia and airway smooth muscle cells probably all interact to increase bronchomotor tone in response to challenge with allergenic stimuli . In addition to important cell-cell interactions, there may be a significant postjunctional component of airway hyperreactivity in which mediators interact at the level of airway smooth muscle cells . To test this hypothesis, I investigated synergistic interactions of methacholine (MCH), histamine and serotonin using intact and chemically skinned canine tracheal smooth muscle . A threshold concentration of histamine (100 nM) significantly increased force when it was added 20 min after or 10 min before stimulation with low concentrations of MCH (1-10 nM) . A threshold concentration of serotonin (10 nM) also significantly increased contractions induced by MCH . Serotonin (10 nM) added during a steady-state contraction induced by 6 or 10 nM MCH increased force and increased fura-2 fluorescence, which suggests that force increases in part because myoplasmic Ca++ increases . I also tested the hypothesis that mechanisms beyond the initial steps of signal transduction contribute to synergism by permeabilizing tracheal smooth muscle fibers with staphylococcal alpha-toxin . Histamine (30 microM), MCH (1 microM), GTP-gamma-S (100 microM) and phorbol dibutyrate (1 microM) all potentiated contractions induced by 1 microM Ca++ in skinned fibers . These data suggest that postjunctional components of airway hyperreactivity may be a combination of synergistic effects of agonists occurring at the level of excitation-contraction coupling as well as sensitization of contractile proteins to Ca++. J Exp Med, 1996 Aug 1, 184(2), 725 - 33 Function of the p55 tumor necrosis factor receptor "death domain" mediated by phosphatidylcholine-specific phospholipase C; Machleidt T et al.; Tumor necrosis factor (TNF) is a pleiotropic mediator of inflammation that has been implicated in the pathogenesis of devastating clinical syndromes including septic shock . We have investigated the role of a TNF-responsive phosphatidylcholine-specific phospholipase C (PC-PLC) for the cytotoxic and proinflammatory activity of TNF . We show here that the cytotoxicity signaled for by the so-called "death domain" of the p55 TNF receptor is associated with the activation of PC-PLC . The xanthogenate tricyclodecan-9-yl (D609), a specific and selective inhibitor of PC-PLC, blocked the cytotoxic action of TNF on L929 and Wehi164 cells . In vivo, D609 prevented both adhesion molecule expression in the pulmonary vasculature and the accompanying leukocyte infiltration in TNF-treated mice . More strikingly, D609 protects BALB/c mice from lethal shock induced either by TNF, lipopolysaccharide, or staphylococcal enterotoxin B . Together these findings imply PC-PLC as an important mediator of the pathogenic action of TNF, suggesting that PC-PLC may serve as a novel target for anti-inflammatory TNF antagonists. Infect Immun, 1996 Aug, 64(8), 3410 - 5 Purification and characterization of the staphylococcal slime-associated antigen and its occurrence among Staphylococcus epidermis clinical isolates; Baldassarri L et al.; The Staphylococcus epidermidis slime-associated antigen (SAA) was purified and characterized . N-Acetyl-glucosamine accounted for 70% of the dry weight of SAA, which was immunolocalized on the ruthenium red-positive material produced by slime-positive strains . A total of 59% of slime-producing S . epidermidis clinical isolates expressed SAA, while the phenotype slime- SAA+ was never recovered. J Immunol, 1996 Aug 1, 157(3), 1200 - 6 Complement activation by a B cell superantigen; Kozlowski LM et al.; Staphylococcal protein A (SpA), acting as a B cell superantigen, binds to the Fab region of human VH3+ Igs . Using SpA abrogated of its IgG Fc binding activity (Mod SpA) as a model B cell superantigen, we determined whether such an interaction causes complement activation . Addition of Mod SpA to human serum led to complement consumption and the generation of C3a . To determine whether this complement activation 1) was due to an interaction between VH3+ Igs and the Fab binding site of SpA and 2) proceeded via the classical complement pathway, we tested a panel of monoclonal IgM proteins for the ability to hind C1q following interaction with SpA . C1q binding was restricted to SpA-reactive, VH3+ IgM proteins . To formally determine whether the binding of SpA to the reactive VH3+ IgM proteins led to complement activation, we reconstituted the serum from a hypogammaglobulinemic patient with monoclonal IgM proteins and measured complement consumption and C3a generation following the addition of Mod SpA . We observed complement consumption and C3a production only in Mod SpA-treated serum reconstituted with a VH3+, SpA-binding, IgM protein . Taken together, these results provide compelling evidence that the interaction of the Fab binding site of SpA and VH3+ Igs can lead to complement activation via the classical pathway . This novel interaction may have significant implications for the in vivo properties of a B cell superantigen. Am J Pathol, 1996 Aug, 149(2), 629 - 38 Prelabeled glycoprotein Ib/IX receptors are not cleared from exposed surfaces of thrombin-activated platelets; White JG et al.; The present investigation has re-examined the hypothesis proposing that glycoprotein (GP)Ib/IX receptors for von Willebrand factor are rapidly cleared from exposed surfaces to internal membrane systems after activation of platelets by thrombin in suspension . Platelets were prelabeled with either a polyclonal antibody to GPIb alpha, antiglycocalicin (A-Gl), or a cocktail of two monoclonal antibodies, AP1 and 6D1, exposed to 0.1 or 0.2 U/ml thrombin for 5 or 10 minutes, fixed and stained with Staphylococcus protein A coupled to gold to detect A-Gl or goat anti-mouse IgG bound to gold particles to locate AP1 and 6D1 before or after preparation of frozen thin sections or embedding for plastic thin sections . The frequency and distribution of protein-A-gold markers for GPIb/IX on thrombin-activated platelets viewed in thin plastic sections did not differ from the density on resting platelets stained with A-Gl . Cryosections of A-Gl-prelabeled platelets labeled again on cryosections revealed GPIb present on linings of the open canalicular system of resting and activated platelets, but the density of gold in interior channels and frequency of gold particles on exterior surfaces were not altered by thrombin stimulation . Platelets prelabeled with the cocktail of 6D1 and AP1 and studied in cryosections also failed to reveal uptake of GPIb/IX receptors into the open canalicular system after activation by thrombin . The findings do not support the concept that thrombin causes clearance of GPIb/IX receptors from exterior surfaces to interior membranes of activated platelets. J Infect Dis, 1996 Aug, 174(2), 338 - 45 Superantigen vaccines: a comparative study of genetically attenuated receptor-binding mutants of staphylococcal enterotoxin A; Bavari S et al.; Superantigens exert their pathologic effects by direct binding to major histocompatibility complex (MHC) class II molecules and T cell antigen receptors (TCR), thus circumventing the normal, antigen-specific immune response . A direct link between disease and toxin suggests an excellent opportunity for vaccine intervention . Site-directed mutants of staphylococcal enterotoxin A (SEA) that have attenuated binding to either the TCR or the MHC class II molecule were developed . Both kinds of SEA mutants induced high levels of antibody against SEA when used as vaccines, and the immunized animals were fully protected when challenged with wild type toxin . However, a residual lethality was associated with the attenuated TCR-binding mutant . These results, combined with an understanding of the molecular nature of superantigen and receptor interactions, indicate that targeting MHC class II binding by site-directed mutagenesis will produce the most effective vaccine. Gastroenterology, 1996 Aug, 111(2), 462 - 71 Interferon gamma plays a critical role in T cell-dependent liver injury in mice initiated by concanavalin A; Kusters S et al.; BACKGROUND & AIMS: T cell-dependent liver injury involving endogenous tumor necrosis factor (TNF) alpha can be induced by either concanavalin A in naive mice or by activating anti-CD3 antibody or staphylococcal enterotoxin B in D-galactosamine-sensitized mice . In this study, the role of interferon gamma (IFN-gamma) in these T-cell models was addressed . METHODS: Mice were pretreated with a neutralizing anti-mouse IFN-gamma antiserum before injection of T cell-activating agents . Plasma cytokine and transaminase levels were determined . Apoptotic cell death was assessed by hepatic DNA fragmentation . RESULTS: Anti-IFN-gamma antiserum significantly protected mice from concanavalin A-induced liver injury . Circulating IFN-gamma was completely suppressed, and TNF was reduced by 50% . Recombinant TNF-alpha administered to mice treated with concanavalin A and anti-IFN-gamma antiserum failed to initiate liver injury . Similar results were obtained with recombinant IFN-gamma in concanavalin A-challenged mice under the condition of TNF neutralization . Neither hepatic DNA fragmentation nor release of transaminases was inhibited by anti-IFN-gamma antiserum when liver injury was induced by staphylococcal enterotoxin B or anti-CD3 antibody in D-galactosamine-sensitized mice . CONCLUSIONS: Both TNF as well as IFN-gamma are critical mediators of liver injury in concanavalin A-treated mice, whereas hepatic DNA fragmentation and liver failure in the D-galactosamine models depend only on TNF. J Biotechnol, 1996 Jul 31, 48(3), 241 - 50 Integrated production of human insulin and its C-peptide; Nilsson J et al.; The potential for the development of an integrated process for production of human insulin and its C-peptide in Escherichia coli has been investigated . Human proinsulin was produced intracellularly in E . coli fused to two synthetic IgG-binding domains (ZZ) derived from staphylococcal protein A . High expression levels (3 g/l culture) of the gene product, which accumulated as inclusion bodies, was obtained . Solubilization of inclusion bodies by oxidative sulfitolysis and subsequent renaturation was performed directly after cell lysis and pellet wash . IgG affinity chromatography was used for efficient recovery of pure proinsulin fusion protein in a single step . Monomers of the proinsulin fusion protein constituted approximately 70% . A single step conversion of the fusion protein into insulin and C-peptide by trypsin and carboxypeptidase B treatment was achieved by engineering the junction between proinsulin and its affinity handle, ZZ . Characterization of the cleavage products by reversed phase chromatography (RPC) verified that human insulin and C-peptide were generated and that the ZZ affinity handle was resistant to cleavage . Human insulin and C-peptide were recovered with high yields by preparative reversed-phase high performance liquid chromatography (RP-HPLC) . The potential use of the presented scheme for large-scale production of recombinant insulin and/or its C-peptide is discussed. J Mol Biol, 1996 Jul 26, 260(4), 570 - 87 Accretion of structure in staphylococcal nuclease: an 15N NMR relaxation study; Alexandrescu AT et al.; 15N main-chain dynamics are compared in four forms of staphylococcal nuclease with different stabilities to unfolding: (1) SN-T, the ternary complex of the protein, Ca2+, and the inhibitor thymidine 3', 5'-bisphosphate; (2) SN, the protein in the absence of added ligands; (3) SN-OB, a folded fragment that corresponds to an "OB-fold" subdomain; (4) delta 131 delta, a denatured 131-residue fragment . SN-T exhibits very little internal motion on the nanosecond timescale . In SN, a moderate increase in flexibility is observed for the first three strands of the five-stranded beta-sheet, and for a loop between strands 4 and 5 . In SN-OB, the loops between strands 3 and 4, and between strands 4 and 5, are extremely flexible on the nanosecond timescale . While the beta-sheets of SN-OB and SN have comparable dynamics on the nanosecond timescale, the beta-sheet in SN-OB experiences additional motion on a slower timescale of 330(+/-170) microseconds . We attribute the latter to interconversion between a major folded (> or = 98%) and a minor unfolded (> or = 2%) conformation . In delta 131 delta, the first three strands of beta-sheet experience conformational averaging on the millisecond timescale . Most of the remainder of the polypeptide chain is highly flexible on the nanosecond timescale . When all four forms of nuclease are considered, there is an increase in the proportion of residues with large amplitude internal motions (low order parameters) as the stability of the folded state is decreased . Residues with low order parameters cluster to distinct regions of the chain, and have H alpha chemical shifts and 3JHN-H alpha coupling constants that tend towards "random coil" values . Conversely, a trend towards uniformly high order parameters suggests a consolidation of structure with increasing stability to denaturation. Int J Cancer, 1996 Jul 17, 67(2), 232 - 7 Tumor-infiltrating lymphocytes can be activated in situ by using in vivo activants plus F(ab')2 bispecific antibodies; Thibault C et al.; In vitro-activated T lymphocytes can be retargeted with anti-CD3 x anti-tumor bispecific antibodies (BsAb) to kill tumor cells in vitro and in vivo . The purpose of the present study was to examine the systemic and intra-tumor effects of in vivo T-cell activation with BsAb, staphylococcal enterotoxin B (SEB), and beta-glucan in combination with BsAb as a retargeting agent . CL-62 melanoma cells were injected subcutaneously into C3H/ HeN mice . Mice were subsequently treated with BsAb alone or with SEB or beta-glucan plus BsAb . Fresh splenocytes, lymph-node cells and tumor-infiltrating lymphocytes (TIL) were tested for their proliferative response using incorporation of 3H-thymidine, and for their ability to lyse CL-62 cells in the presence or absence of BsAb in 4-hr 51Chromium release assays . Toxicity of treatments was assessed in a D-galactosamine model . BsAb, alone or combined with beta-glucan, had essentially no effect on systemic T-cell cytotoxicity, and essentially no effect on systemic proliferation, unless exogenous IL-2 was provided . At the tumor site, BsAb alone, BsAb plus beta-glucan, and SEB plus BsAb all significantly increased BsAb-mediated TIL cytotoxicity . In contrast to the TIL-limited effects of BsAb and of BsAb plus beta-glucan, SEB plus BsAb markedly increased both systemic and intra-tumor T-lymphocyte activation . Toxicity correlated with measures of systemic activation, with limited effects from BsAb alone and from beta-glucan plus BsAb, and with marked lethality from SEB plus BsAb . Overall, these results suggest moderate intra-tumor activation of TIL, but no measurable systemic activation after in vivo treatment with BsAb or beta-glucan plus BsAb . SEB plus BsAb results in complete T-cell activation in both systemic and intra-tumor compartments, but at the expense of increased systemic toxicity . In conclusion, TIL can be activated in situ with different combinations of in vivo activants . In vivo-activated TIL can be retargeted with bispecific antibodies to lyse tumor cells, and may be an alternative to ex vivo T-cell activation and adoptive transfer therapy. J Mol Biol, 1996 Jul 5, 260(1), 22 - 33 Sequence-specific DNA binding of the phage Mu C protein: footprinting analysis reveals altered DNA conformation upon protein binding; Ramesh V et al.; The mom gene of bacteriophage Mu, which codes for a DNA modification function, is regulated in a complex manner at both transcriptional and translational levels . The phage-encoded C protein functions as an activator of mom transcription . The mom promoter has features of an activator-dependent weak promoter, and the C binding site is located upstream and overlapping the -35 region and includes the palindromic sequence TTAT(N)6ATAA . The interactions of this activator protein at its binding site in Pmom has been investigated using four different chemical footprinting reagents . The protein footprint spans a region of 18 to 25 bp, depending on the nature of the chemical reagent used . Dimethylsulfate protection experiments revealed the base-specific interactions . The protected guanines are separated by 15 bp and are located beyond the interrupted palindromic sequence . A tripartite footprint was observed with hydroxyl radical, generated by Fe(II)-EDTA, which shows the binding of the protein to one face of the helix . The extent of protection conferred by the bound protein, however, is not uniform, suggesting that the interaction is asymmetric . The chemical nuclease 1,10-phenanthroline-copper, a minor groove specific ligand, shows hyper-reactivity upon protein binding in the top strand nucleotide triplet CAC, again confirming the protein-induced alterations in DNA conformation . Gel exclusion chromatography and chemical crosslinking experiment with the purified protein suggest that this mode of interaction is accomplished by a dimeric protein . This observation is supported by electrophoretic mobility shift assay using heterodimer of pure C protein and staphylococcal protein A-C fusion . The deletion analysis implicates a role for the carboxyl-terminal region of the protein in DNA binding. Rom J Morphol Embryol, 1996 Jul-Dec, 42(3-4), 169 - 78 Anatomoclinical and bacteriological study on chronic marginal periodontal disease . Therapy with staphylococcic vaccine; Laky D et al.; Histological, histochemical and electronmicroscopic investigations were carried out on gum biopsies taken from cases with chronic marginal periodontal disease before and after the treatment with staphylococcic vaccine, thus, morphopathologically proving the findings of Gafar et al . (1985) and of Georgescu et al . (1973; 1974; 1975; 1979; 1995) . Before treatment, gum lesions were severe, consisting in an abundance of polymorphic inflammatory infiltrates in the chorion along with oedema and disorganised connective fibres, atrophies and ulceration of the mucosa, purulent deposits at the surface with bacteria and fungi . After the treatment with the staphylococcic vaccine, an epithelial regeneration was seen together with the absence of the chronic inflammation . The intensely fibroparius granulated tissue was also noted, in all cases observing abundant connective fibres, accounting for good results after treatment, notably the firmness of the tooth fitting. Microb Drug Resist, 1996 Summer, 2(2), 239 - 43 An electron microscopic study of clinical and laboratory-derived strains of teicoplanin-resistant Staphylococcus haemolyticus; Giovanetti E et al.; Staphylococcal resistance to glycopeptides (which involves more teicoplanin than vancomycin) is uncommon and largely confined to Staphylococcus haemolyticus, an emerging nosocomial pathogen with a tendency to develop antibiotic resistance . In this study, six S . haemolyticus strains, including two isogenic pairs of teicoplanin-susceptible/-resistant strains and two resistant clinical isolates, were used in a morphologic and morphometric electron microscope investigation . Cells from both clinical and laboratory-derived teicoplanin-resistant strains exhibited abnormally roughened, irregular outlines when observed by transmission electron microscopy . However, no significant differences in cell wall thickness resulted from morphometric analysis when the susceptible/resistant cells of the two isogenic pairs were compared . By scanning electron microscopy, an abnormally roughened, blistered surface was associated with teicoplanin-resistant cocci . A certain variability was noted between strains, not clearly related to the resistance level . In freeze-fracture investigations, a higher number per square micrometer of intramembrane particles, more significant in the E than in the P membrane fracture face, was observed in the laboratory-derived resistant clones as compared to susceptible parent strains . Further studies are needed to understand the cause-effect relation between these ultrastructural alterations and staphylococcal resistance to teicoplanin (but not to vancomycin). Zh Mikrobiol Epidemiol Immunobiol, 1996 Jul-Aug, (4), 71 - 4 {An immunomodifier--staphylococcal anatoxin--prevents the development of immunosuppression caused by informational stress in mice}; Ozherelkov SV et al.; The action of information stress for 14 days leads to the development of immunosuppression, which is manifested by the suppression of humoral response to sheep red blood cells (SRBC) and the decrease of resistance to Langat virus having low pathogenicity . As shown in this investigation, an immunomodifier, purified staphylococcal toxoid (PST), protects experimental animals from the immunosuppressive effect of information stress . After the injection of PST to stress-affected mice in doses of 15 or 1.5 binding units per animal on days 9, 11 and 13 of the experiment their humoral response to SRBC and resistance to Langat virus are partially restored (by 45-60%). Izv Akad Nauk Ser Biol, 1996 Jul-Aug, (4), 499 - 501 {The effect of long-term UV radiation exposure on the skin autonomous microflora and on body nonspecific resistance}; Strzhizhovskii AD et al.; Hairless mice of strain HRS were exposed to vertical UV-B-irradiation (280-320 nm) at daily doses of 1 or 2 kJ/m2 (biologically effective doses 200 and 400 J/m2) five days a week for 13 and 6 weeks . As the length of exposure increased, the abundance of automicroflora of the dorsal skin first decreased, then exceeded the control value, was normalized, and remained at the normal level until the end of the experiment . The rates of effect appearance and disappearance, just as its amplitude, increased with the daily doses . No changes were foun