Biochemistry, 1992 Oct 27, 31(42), 10217 - 25 Compact denatured state of a staphylococcal nuclease mutant by guanidinium as determined by resonance energy transfer; James E et al.; The protein from a mutant clone of staphylococcal nuclease with a cysteine substituting for a lysine at position 78 was prepared and labeled with a cysteine-specific fluorescent probe 5-{{2-{(iodoacetyl)-amino}ethyl}amino}naphthalene-1-sulfonic acid (IAEDANS) . Time-resolved nonradiative energy-transfer studies were done using the single tryptophan at position 140 as the energy donor and the IAEDANS as the receptor . Changes in distance and distance distributions were observed as a function of increasing guanidinium (GuHCl) concentration (0-2 M) and in the presence or absence of Ca2+ and inhibitor 2'-deoxythymidine 3',5'-diphosphate (pdTp) . In the native state, both the ternary complex and the noncomplexed protein are best fit with one population having an average donor-acceptor distance of approximately 23 A and an "apparent" full width at half-maximum (fwhm) of distance distribution of approximately 18 A . Besides the contribution of linker arm of the acceptor, it appears that there are some conformational heterogeneties either due to the disordering of the tryptophan region or due to the whole protein in the native state . During GuHCl unfolding, the average distance remains relatively constant up to GuHCl concentrations where both the ternary complex and the ligand-free protein are denatured (1-1.3 M) . The compact denatured states persist up to 2 M GuHCl . At 2 M GuHCl, the heterogeneity of the denatured state in the ternary complex is much larger than that of the ligand-free nuclease . The results show that the denatured states of staphylococcal nuclease mutant K78C by GuHCl are compact and these compact denatured states are likely due to residual structures or incompletely disrupted hydrophobic cores under these conditions. J Biol Chem, 1992 Oct 25, 267(30), 21297 - 9 Staphylococcal ADP-ribosyltransferase-sensitive small G protein is involved in brefeldin A action; Sugai M et al.; An early event in the action of brefeldin A (BFA) is the dissociation of beta-coat protein (beta-COP) from the Golgi membrane . We have recently shown that staphylococcal ADP-ribosyltransferase (epidermal cell differentiation inhibitor (EDIN)), which specifically modifies a small G protein, rho, mimics the action of BFA and disassembles the Golgi apparatus in Vero cells (Sugai, M., Chen, C-h., and Wu, H . C . (1992) Proc . Natl . Acad . Sci . U.S.A . 89, 8903-8907) . Three independent BFA-resistant cell lines (BER-40 from Vero cells, PtK1, and MDCK) showed cross-resistance to EDIN regarding the release of the beta-COP from the Golgi membrane by EDIN or BFA . BFA as well as EDIN induced disassembly of the actin microfilaments in Vero cells, and they both failed to induce the disassembly of actin microfilaments in BER-40, PtK1, and MDCK cells . BFA inhibited protein secretion in Vero cells but not in BFA-resistant cell lines, whereas EDIN did not inhibit protein secretion in either Vero or other cell lines . AlF-4 inhibited the effect of EDIN as well as that of BFA on the distribution of the beta-COP . These results suggest that an EDIN-sensitive rho protein together with trimeric and other small G protein(s) is involved in the regulation of the assembly of coated vesicles and vesicular transport in the Golgi apparatus. Proc Natl Acad Sci U S A, 1992 Oct 15, 89(20), 9657 - 61 Distinct binding sites on HLA-DR for invariant chain and staphylococcal enterotoxins; Karp DR et al.; During biosynthesis, class II molecules of the major histocompatibility complex exist as complexes of the polymorphic alpha and beta chains in association with trimers of the invariant chain (Ii) . The nonpolymorphic Ii contains sequences necessary for proper targeting of class II to endosomal compartments, where Ii is degraded . Ii also prevents the premature association of antigenic peptides with class II molecules . It is not known whether the effect of Ii on peptide binding extends to other ligands of class II, specifically exogenous superantigens . Cells expressing a mutant Ii molecule stably associated with HLA-DR at the cell surface were tested for their ability to interact with staphylococcal toxins . Most toxins (staphylococcal enterotoxins A-E and exfoliative toxin) were found to bind to cells expressing this alpha beta Ii complex with levels comparable to cells expressing only alpha beta chains at the cell surface . Cells expressing surface alpha beta Ii complexes stimulated polyclonal populations of peripheral blood T cells in association with these toxins . Binding of toxic shock syndrome toxin (TSST) and subsequent stimulation of T cells were reduced by the presence of the Ii . This reduction was not due to an alteration in the repertoire of T cells responding to TSST in the presence of Ii . Data from experiments with a T-cell clone suggest that interactions between class II molecules and T-cell antigen receptors occur during staphylococcal enterotoxin-mediated stimulation and that surface Ii does not interfere with such interactions. Cancer Res, 1992 Oct 15, 52(20), 5775 - 9 Phase II trial of suramin in patients with advanced renal cell carcinoma: treatment results, pharmacokinetics, and tumor growth factor expression; Motzer RJ et al.; Twenty-six patients with advanced renal cell carcinoma were treated with suramin administered by continuous infusion, with dosing determined by a nomogram . One patient achieved a partial response and five patients achieved a minor response or had stable disease for > 3 months . Toxicities included an immune-mediated thrombocytopenia in one patient and Staphylococcus sepsis that was not associated with neutropenia in five patients . Pharmacokinetic parameters were determined by the ADAPT II MAP-Bayesian parameter estimation program . Patient data were fit using a two-compartment open model and first-order rate elimination . This showed a wide interpatient variation in time to target level (median, 13.8 days), volume of distribution (median, 15.2 liters/m2), and t1/2-beta (median, 20.6 days) . The patients who achieved a partial response, minor response, or stable disease had a slower elimination rate of suramin, compared to patients with progressive disease . Tumor specimens were obtained prior to therapy and were analyzed for the production of five different growth factor-specific RNA transcripts . These included transforming growth factor alpha, acidic fibroblast growth factor, basic fibroblast growth factor, and platelet-derived growth factor types A and B . No difference in the pattern of growth factor expression was seen in tumors of responding and nonresponding patients . Suramin does not have significant antitumor activity in renal cell carcinoma . The wide variability in pharmacokinetics suggests that individual dosing should be used in future trials of suramin for treatment for other malignancies . Pertinent corollary studies of tumor biology and clinical pharmacology should be included whenever possible in clinical trials in patients with renal cell carcinoma. Biochemistry, 1992 Oct 13, 31(40), 9665 - 72 Three-dimensional solution structure of the B domain of staphylococcal protein A: comparisons of the solution and crystal structures; Gouda H et al.; The three-dimensional solution structure of the recombinant B domain (FB) of staphylococcal protein A, which specifically binds to the Fc portion of immunoglobulin G, was determined by NMR spectroscopy and hybrid distance geometry-dynamical simulated annealing calculations . On the basis of 692 experimental constraints including 587 distance constraints obtained from the nuclear Overhauser effect (NOE), 57 torsion angle (phi, chi 1) constraints, and 48 constraints associated with 24 hydrogen bonds, a total of 10 converged structures of FB were obtained . The atomic root mean square difference among the 10 converged structures is 0.52 +/- 0.10 A for the backbone atoms and 0.98 +/- 0.08 A for all heavy atoms (excluding the N-terminal segment from Thr1 to Glu9 and the C-terminal segment from Gln56 to Ala60, which are partially disordered) . FB is composed of a bundle of three alpha-helices, i.e., helix I (Gln10-His19), helix II (Glu25-Asp37), and helix III (Ser42-Ala55) . Helix II and helix III are antiparallel to each other, whereas the long axis of helix I is tilted at an angle of about 30 degrees with respect to those of helix II and helix III . Most of the hydrophobic residues of FB are buried in the interior of the bundle of the three helices . It is suggested that the buried hydrophobic residues form a hydrophobic core, contributing to the stability of FB.(ABSTRACT TRUNCATED AT 250 WORDS) Br J Hosp Med, 1992 Oct 21-Nov 3, 48(8), 496 - 7, 500-3 Infections and intravascular devices; Elliott TS et al.; Complications associated with intravascular devices include infections mainly caused by Staphylococcus epidermis and S . aureus . The reported incidence of these infections varies . Several factors influence the propensity for catheter infections . We recommend strategies for the prevention and treatment of catheter-related sepsis. Am J Cardiol, 1992 Oct 1, 70(9), 925 - 9 Transesophageal echocardiographic diagnosis of right-sided cardiac masses in patients with central lines; Cohen GI et al.; Transesophageal echocardiography provides excellent images of intracardiac masses; however, its use among a series of patients with central venous lines has not been fully described . Nineteen patients (aged 52 +/- 16 years; 10 women) had masses detected by transesophageal echocardiography in the presence of a permanent (0.2 to 16 years) pacing wire (n = 8), and a current (n = 9) or recent (n = 2) (1 to 281 days) indwelling catheter . Transthoracic echocardiography suggested the presence of a mass in 5 patients (26%), although in only 2 cases were its findings consistent with transesophageal findings . Transesophageal echocardiography indicated the presence of a mass in or near the superior vena cava in 13 patients, in the right atrium in 6, and adjacent to the tricuspid valve in 3 . Discrete masses measured 1.6 +/- 2.1 cm2 in area during transesophageal echocardiography . Eleven patients had positive blood cultures, 7 with staphylococcal species . Mass size was not significantly altered by the type of line or sepsis, but showed a weak correlation with line age (r = 0.56) . Transesophageal echocardiography altered the management of 9 patients, prompting surgery (n = 3) and line removal (n = 3), and antibiotic (n = 2) or anticoagulation (n = 3) therapy. J Infect Dis, 1992 Oct, 166(4), 861 - 5 The use of nonsteroidal antiinflammatory drugs to prevent adherence of Staphylococcus epidermidis to medical polymers; Farber BF et al.; The effect of salicylates and other nonsteroidal antiinflammatory drugs on the production of Staphylococcus epidermidis extracellular slime was studied . A dose-related decrease in slime production was observed with increasing concentrations of salicylic acid . S . epidermidis grown in 5 mM salicylic acid were less likely to adhere to Silastic, polyvinyl chloride, polyurethane, and Teflon catheters (P less than .006); strains grown in 2 mM salicylic acid, ibuprofen, indomethacin, or phenylbutazone were less adherent to Silastic catheters (P less than .001) . Similar results were obtained with polyurethane catheters . S . epidermidis strains were less likely to adhere (43%-82% inhibition) to polyurethane catheters treated with 500 mM salicylic acid diluted in ethanol (P less than .0001) . Similar differences were not observed with acetaminophen, ibuprofen, or acetylsalicylic acid . Adherence of radiolabeled S . epidermidis to salicylic acid-treated Silastic catheters demonstrated a dose-related reduction . The use of salicylic acid to coat medical devices may decrease the incidence of device-related infection. Minerva Cardioangiol, 1992 Oct, 40(10), 375 - 81 {Intraoperative microbiological monitoring in abdominal aortic aneurysms in elective surgery . A review of the literature and the authors' personal experience}; Raso AM et al.; Thirty-one patients, ranging in age from 57 to 78 years (mean 66), with the exclusion of cases with doubtful possible results, underwent abdominal aortic aneurysmectomy for asymptomatic AAA and had cultures from the aneurysmal wall and endovascular thrombus to identify possible microbiological source of future graft infection; 5 (16%) of 31 cultures yielded bacterial growth and the most common organism isolated was staphylococcus epidermidis . During an average follow-up of 15.4 months no graft infection was noted in patients with positive or negative aortic cultured . A literature review stresses the same disparity between positive cultures obtained at the aneurysmectomy and subsequent low graft-infection rate . It is concluded that the aneurysm wall itself does not represent an important source of early or late graft infection and it's suggested that the bacterial presence both in the wall and thrombus could be explained by an exogenous contamination at the operation time. Zentralbl Bakteriol, 1992 Oct, 277(3), 357 - 63 Thymocyte proliferation and maturation in response to staphylococcal lipoteichoic acid; Ohshima Y et al.; Lipoteichoic acid (LTA) from Staphylococcus saprophyticus strain S1 could be shown to induce thymocyte proliferation and maturation in BALB/c-mice after systemic administration . The increase in thymocyte numbers per mg organ weight was statistically significant . Determination of thymic lymphatic subsets revealed a considerable up-regulation of mature cells expressing helper/inducer (L3T4) or cytotoxic/suppressor (Lyt-2) phenotypes . Thus administration of staphylococcal LTA obviously accelerated murine thymocyte proliferation and maturation . Counts of BALB/c-mouse peripheral blood lymphocytes (PBL) revealed no evident fluctuation within one week after LTA administration, however, statistically significant increases could be detected two weeks after treatment . The determination of activated PBL expressing IL-2 receptors suggested that injection of staphylococcal LTA apparently induced an immunostimulation since those cells were significantly enhanced within one week after LTA administration. Thorac Cardiovasc Surg, 1992 Oct, 40(5), 261 - 5 Total correction of tetralogy of Fallot in adolescents and adults; Lukacs L et al.; From 1976 to 1988, 23 adolescent and adult patients underwent total correction of tetralogy of Fallot . There were 13 males and 10 females, ranging in age from 16 to 47 years (mean 24.3 +/- 8.6 years) . Eight patients were in New York Heart Association (NYHA) functional class II, 14 patients in class III, and one patient in class IV . Sixteen patients (69.6%) had undergone previous palliative operation . All shunts were patent at the time of repair . In 9 patients bovine pericardial monocusp patches were used for reconstruction of the right-ventricular outflow tract . Intraoperatively, the right-ventricular to left-ventricular systolic pressure ratio after repair ranged from 0.29 to 0.80 (mean 0.49 +/- 0.13) . There were 2 early deaths (8.7%) . Eight of 23 patients (34.8%) exhibited postoperative low cardiac output syndrome . One late death occurred: a 22-year-old male patient died of Staphylococcus sepsis 8 months postoperatively . All surviving patients were followed from 3 to 15 years (mean 8.3 +/- 2.7 years) . No patient required reoperation in the follow-up period . The actuarial survival estimate for all 23 patients was 87% at the end of 15 years . At follow-up 17 patients were in NYHA class I, two were in class II, and one was judged to be in class III . We believe advanced age is no contraindication to surgery in tetralogy of Fallot . Adolescents and adults remain in need of total correction which can be performed with acceptable risk and long-term symptomatic improvement. Can J Vet Res, 1992 Oct, 56(4), 382 - 6 The use of an implantable central venous (Hickman) catheter for long-term venous access in dogs undergoing bone marrow transplantation; Abrams-Ogg AC et al.; Methods were developed for the insertion and maintenance of long-term central venous catheters in dogs in order to provide reliable venous access during bone marrow transplantation . Single-lumen, 9.6 Fr Hickman catheters with a VitaCuff were used . The catheter was inserted into the jugular vein via a surgical cut-down, and tunnelled subcutaneously to exit over the thoracic spine . Fluoroscopic guidance was necessary to ensure proper positioning of the catheter tip in the right atrium . The catheter was secured at the venous entrance site with a grommet and at the cutaneous exit site with a finger-cuff suture . The exit site was bandaged; dressings were changed daily . Five dogs were studied . Catheter insertion and maintenance techniques were developed using two dogs . For the other three dogs, which developed 7 wk of profound myelosuppression induced by total body irradiation, the catheters were used for blood sampling and infusions of antibiotics, fluids, and blood products . For these three dogs there were 261 total catheter-days . Complete catheter obstruction did not occur . Partial obstruction (inability to withdraw blood) occurred for 13 days with one catheter . The tip of this catheter was in the cranial vena cava . One irradiated dog had a staphylococcal exit site infection for several days after catheter insertion, which resolved with antibiotic therapy . Infections of the subcutaneous tunnel, and catheter associated bacteremia, were not identified . Infectious and hemorrhagic complications of myelosuppression were less severe than in six other dogs where intermittent venipuncture was used for vascular access during radiation induced myelosuppression . In conclusion, long-term central venous catheterization is feasible in dogs during profound myelosuppression and markedly facilitates patient management. J Rheumatol, 1992 Oct, 19(10), 1586 - 90 Subcutaneous bursitis in scleroderma; Lagana A et al.; We encountered 3 types of subcutaneous bursitis in our patients with scleroderma: dry bursitis characterized by a rub, sterile bursitis characterized by inflammatory effusions without crystals by polarizing microscopy, and septic (staphylococcal) subcutaneous bursitis . The latter, which occurred in 6 of 40 consecutive patients, had a protracted course, was often complicated by fistulas, and tended to involve several bursae particularly in patients with extensive calcinosis. Anal Biochem, 1992 Oct, 206(1), 12 - 6 Fluorimetric assay of D-lactate; McLellan AC et al.; A fluorimetric assay for D-lactate in human blood samples was developed using an endpoint enzymatic assay with D-lactate dehydrogenase from Staphylococcus epidermidis . The intrabatch and interbatch coefficients of variance were 8.7% (n = 4) and 16.6% (n = 4), respectively . The limit of detection in blood was 3.73 nmol/ml . The assay suffers minor interference from S-D-lactoylglutathione, which was also present in the blood samples . The concentration of D-lactate in blood was (mean +/- SE, nmol/ml) normal healthy individuals, 11.0 +/- 1.2 (n = 7); and diabetic patients, 20.0 +/- 1.3 (n = 55) (a significant increase in diabetes mellitus; P < 0.01, Mann-Whitney U test). Int J Immunopharmacol, 1992 Oct, 14(7), 1285 - 92 Peptides and their constituent amino acids influence the immune response and phagocytosis in different ways; Belokrylov GA et al.; To elucidate the role of immunoactive amino acids (which are capable of stimulating the immune response) in the peptide regulation of antibody production and phagocytic processes we have studied the influence of some fragments of natural peptides and the amino acids included in them on the thymus-dependent immune response to sheep red blood cells (SRBC) and on the phagocytosis of Staphylococcus by murine peritoneal neutrophils . It has been found that the effects of amino acids, as well as of peptides that they form, on the immune response and on phagocytosis were diverse . Glutamic and aspartic acids, threonine and valine stimulated both the immune response and phagocytosis . Glycine and isoleucine influenced neither the immune response nor phagocytosis, whereas lysine, proline, tyrosine and leucine did not influence the immune response, but enhanced the phagocytic activity of neutrophils . Arginine inhibited the immune response but stimulated phagocytosis . Peptide TTKD section (the fragment 77-80 of murine Thy-1-antigen) contained in C- and N-terminus amino acids (T and D) which stimulated the antibody production and phagocytosis, and lysine (K) which stimulated phagocytosis only, enhanced both processes . Peptides LGIP and PYIK (the fragments 49-53 and 66-69 of murine Thy-1 antigen) which contained both immunologically inert amino acids (L, G, I, P, Y, K) and phagocytosis stimulating amino acids (L, Y, P, K) influenced neither the immune response nor the phagocytic activity of neutrophils.(ABSTRACT TRUNCATED AT 250 WORDS) Ginecol Obstet Mex, 1992 Oct, 60, 272 - 6 {Experience with the management of endometritis in the Instituto Nacional de Perinatalogía}; Figueroa Damian R et al.; Endometritis is the most frequent infectious complication of the puerperal period; with the objective to know the clinical characteristics, etiology and evolution of this disease we did a retrospective study of the endometritis cases among the patients of the Instituto Nacional de Perinatologia (INPer) from January 1st, 1990 to May 31, 1991 . During the revision period were diagnosed 120 cases, but we were able to examine only 90 . In the 90% the resolution of the pregnancy was by means of caesarean . A 24.4% of the patients had premature rupture of the membranes . The 10% had diagnose of chorioamniotis . The latency period to develop postpartum endometritis was 120 +/- 66 hours, and post-caesarean 56 +/- 30 hours (p = 0.001) . The symptom most frequent were fever (100), foul-smelling lochia (61.1%) and uterine tenderness (60%) . The infection was polymicrobial, being the microorganisms most frequent isolated Staphylococcus coagulase negative, Escherichia coli and Peptostreptococcus . A 3.3% developed pelvi-peritonitis . One patient died. J Exp Med, 1992 Oct 1, 176(4), 1063 - 72 Defective maintenance of T cell tolerance to a superantigen in MRL-lpr/lpr mice; Zhou T et al.; In normal mice neonatal injection of staphylococcal enterotoxin B (SEB) induces tolerance in T cells that express reactive T cell receptor (TCR) V beta regions . To determine if a T cell neonatal defect was present in MRL-lpr/lpr mice, 20 micrograms of SEB was injected intraperitoneally every other day into V beta 8.2 TCR transgenic and nontransgenic MRL(-)+/+ and MRL-lpr/lpr mice from birth to 2 wk of age . At 2 wk of age, V beta 8+ T cells were depleted, and SEB reactivity was lost, in spleen, lymph node, and thymus . These effects were equivalent in +/+ and lpr/lpr SEB-tolerized mice . However, MRL-lpr/lpr mice failed to maintain neonatal tolerance . By 4 wk of age, there was a dramatic increase in T cells expressing V beta 8.2 in the peripheral lymph nodes of MRL-lpr/lpr mice but not MRL(-)+/+ mice . In vitro stimulation with SEB or TCR crosslinking revealed a total loss of neonatal tolerance 2 wk after cessation of SEB treatment in lpr/lpr mice, but not +/+ mice . The time-course of recovery of V beta 8+ T cells and reactivity to SEB and TCR crosslinking in the thymus of MRL-lpr/lpr mice was similar to that in the lymph node . Thymectomy at 2 wk of age eliminated tolerance loss in lymph nodes of MRL-lpr/lpr mice at 4 wk of age, indicating that loss of peripheral tolerance was due to the emigration of untolerized T cells from the thymus . Challenge of neonatally tolerized MRL-lpr/lpr mice with SEB (100 micrograms, i.p.) at 8 wk of age resulted in a dramatic onset of T cell-mediated autoimmune disease characterized by 30% weight loss and 60% morality . This indicated that loss of tolerance to SEB also occurred in vivo . In contrast, neonatally tolerized MRL(-)+/+ mice remained totally unresponsive to SEB challenge and did not undergo any detectable weight loss . These results suggest that there is normal induction of neonatal tolerance to SEB in lpr/lpr mice, but that tolerance is not maintained after the tolerizing antigen is removed . This loss of neonatal tolerance can lead to severe weight loss and death on exposure to the tolerizing antigen later in life. J Leukoc Biol, 1992 Oct, 52(4), 469 - 72 A synthetic peptide homologous to retroviral envelope protein down-regulates TNF-alpha and IFN-gamma mRNA expression; Haraguchi S et al.; We investigated the influence of CKS-17, a synthetic heptadecapeptide that corresponds to a highly conserved domain of the immunosuppressive retroviral envelope protein p15E, on staphylococcal enterotoxin B (SEB)-induced TNF-alpha gene expression in human peripheral blood mononuclear cells and highly purified human monocyte preparations, as well as the production of TNF-alpha protein, using human peripheral blood mononuclear cells . RNA hybridization studies show that CKS-17 inhibits SEB-induced TNF-alpha mRNA accumulation in human peripheral blood mononuclear cells and human monocytes . CKS-17 is also shown to be highly suppressive for SEB-induced production of TNF-alpha proteins . Similarly, CKS-17 inhibits expression of SEB-induced IFN-gamma mRNA in human peripheral blood mononuclear cells . These results suggest that CKS-17 down-regulates both TNF-alpha and IFN-gamma production at mRNA level. Infect Immun, 1992 Oct, 60(10), 4322 - 7 Hemagglutination and adherence to plastic by Staphylococcus epidermidis; Rupp ME et al.; Staphylococcus epidermidis is an important nosocomial pathogen responsible for intravenous catheter-related bacteremia and infections of other prosthetic medical devices . We found that the ability of S . epidermidis to hemagglutinate erythrocytes correlated with the adherence of bacteria to plastic and to intravenous catheters . S . epidermidis isolates responsible for prosthetic-valve endocarditis (n = 61) and isolates from intravenous catheters (n = 59) were significantly more likely to cause hemagglutination than isolates from the skin of preoperative cardiac surgery patients (n = 19) (P = 0.027) . S . epidermidis isolates (n = 23) recovered from the skin of patients 7 to 10 days after cardiac surgery were significantly more likely to exhibit hemagglutination than the preoperative isolates (P = 0.015) . By a quantitative adherence assay, we also observed that the hemagglutination titer and number of species of erythrocytes agglutinated correlated directly with adherence to polystyrene (P less than 0.001) . In addition, hemagglutinating isolates were significantly more likely to be recovered in high number from intravenous catheters when semiquantitative catheter culture techniques were used (P less than 0.001) . We speculate that hemagglutinin(s) either plays a direct role in adherence to polymers and thus prosthetic-device infection or serves as an easily demonstrable marker for adherence-prone isolates. Infect Immun, 1992 Oct, 60(10), 4127 - 32 Staphylococcus saprophyticus hemagglutinin is a 160-kilodalton surface polypeptide; Gatermann S et al.; Many strains of Staphylococcus saprophyticus cause direct hemagglutination of sheep erythrocytes . For a high proportion of clinical isolates, a surface protein (Ssp) that is apparently not involved in this property has been described . In this study, S . saprophyticus CCM883, a hemagglutinating but Ssp-negative strain, was used for the identification, purification, and characterization of a 160-kDa surface polypeptide that appears to be the major component of the hemagglutinin . Expression of the protein required the addition to the growth medium of EDTA in micromolar quantities, suggesting an inhibitory role for some unidentified metal ion . The protein was purified by means of Sephacryl S-300 chromatography, and antisera were raised in rabbits . Antibody against this protein inhibited the hemagglutination of two other, unrelated strains and was used to demonstrate, by electron microscopy, the presence of the protein on the surface of the cells . In a confirmatory experiment, the purified antigen was incubated with erythrocytes and binding was detected by the Western immunoblot technique with the antibody to the 160-kDa polypeptide . These experiments indicate that this surface protein is the hemagglutinin of S . saprophyticus. Eur J Immunol, 1992 Oct, 22(10), 2749 - 52 A versatile method to produce antibodies to human T cell receptor V beta segments: frequency determination of human V beta 2+ T cells that react with toxic-shock syndrome toxin-1; Romagne F et al.; Human V beta (hV beta) regions have been expressed in the context of mouse T cell receptor (TcR)-CD3 complexes, and subsequently used to raise hV beta-specific monoclonal antibodies (mAb) . The method of expression of hV beta outlined in this report contrasts in its versatility with the one reported by Choi et al . (Proc . Natl . Acad . Sci . USA 1991 . 88: 8357) . For instance, we have applied it successfully to the construction of mouse T cell transfectants expressing hV beta 1, hV beta 2, hV beta 3, hV beta 8, hV beta 9, hV beta 13.5, hV beta 19, hV beta 21, and hV beta 22 gene segments . mAb against the hV beta 2 and hV beta 19 regions have been raised by using these transfectants as immunogens in mice . Here, we illustrate the application of the anti-hV beta 2 mAb to the measurement of human T cells that react with the staphylococcal toxic-shock syndrome toxin-1. Lett Appl Microbiol, 1992 Oct, 15(4), 142 - 5 Plasmid-encoded resistance to tetracycline in Staphylococcus haemolyticus; Schwarz S; A small 4.35 kb plasmid, designated pSTS4, was isolated from a multiresistant Staphylococcus haemolyticus culture . It conferred resistance to tetracycline as shown by protoplast transformation . pSTS4 was further characterized by restriction endonuclease analyses and a preliminary restriction map constructed . It shared some structural similarities with previously described small TcR plasmids from other staphylococcal species of human and animal origin . pSTS4 encoded two proteins of approximately 37 kDa and 52 kDa as revealed by combined in vitro transcription/translation assays. J Chemother, 1992 Oct, 4(5), 281 - 5 Piperacillin/tazobactam plus gentamicin as empirical therapy for febrile neutropenic patients with haematological malignancy; Kelsey SM et al.; The efficacy of piperacillin/tazobactam (PIPC/TBT) in combination with gentamicin was assessed as empirical therapy in 44 febrile neutropenic patients with haematological malignancy . A favourable response to therapy was seen in 67% patients overall and in 57% of patients with microbiologically documented infection . PIPC/TBT demonstrated good clinical and in vitro activity against isolated pathogens, particularly Gram positive cocci such as Staphylococcus epidermidis . The MIC of both Gram positive and Gram negative pathogens to PIPC was reduced in the presence of TBT . PIPC/TBT plus gentamicin is a safe and effective combination for empirical therapy in febrile neutropenic patients, even in a unit with a predominance of Gram positive infections. Circ Res, 1992 Oct, 71(4), 951 - 9 Endothelin increases myofilament Ca2+ sensitivity in alpha-toxin-permeabilized rabbit mesenteric artery; Nishimura J et al.; This study was designed to investigate the mechanism of endothelin-1 (ET-1) contractions in Staphylococcus alpha-toxin-permeabilized vascular smooth muscle . Rabbit small mesenteric arteries permeabilized with alpha-toxin were mounted for isometric or isotonic force recording or were processed for determination of myosin light chain (MLC) phosphorylation levels . Addition of 100 nM ET-1 plus 10 microM GTP significantly enhanced myofilament Ca2+ sensitivity as compared with the addition of Ca2+ alone (EC50, 0.47 microM Ca2+ for Ca2+ alone and 0.13 microM Ca2+ for ET-1 plus (GTP) . This enhanced sensitivity was reversed by GDP beta S . ET-1-induced contractions were relaxed at a constant {Ca2+} by the addition of 30 microM cAMP or cGMP, demonstrating a direct effect of the cyclic nucleotides on contractile regulation . Inhibition of protein kinase C activity by 100 nM staurosporine relaxed ET-1 plus GTP-induced contractions, and pretreatment with 40 microM chelerythrine inhibited the ET-1 plus GTP increase in force . At 0.32 microM Ca2+, steady-state levels of shortening velocity were not increased by ET-1 plus GTP, although steady-state levels of MLC phosphorylation were significantly enhanced . The ET-1-induced increase in MLC phosphorylation was not altered by changes in {Ca2+}, whereas the shortening velocity was Ca2+ dependent, suggesting that the increase MLC phosphorylation level may be the result of protein kinase C, rather than MLC kinase, activation . These results are consistent with the hypothesis that ET-1 increases myofilament Ca2+ sensitivity by a G protein-dependent pathway and subsequent activation of protein kinase C.(ABSTRACT TRUNCATED AT 250 WORDS) Indian J Med Sci, 1992 Oct, 46(10), 297 - 300 Manage of acute osteomyelitis in children--should it be conservative? Sadat-Ali M. 73 children with acute osteomyelitis seen at King Fahd University Hospital, Al-Khobar, Saudi Arabia were reviewed . Majority of patients were between the 6-10 year age group and male children predominated . All patients had incision and drainage and antibiotic therapy for a minimum of 6 weeks . Metaphysis of the tibia was the commonest site of infection and the staphylococcal species was cultured in 77% of children . Ampiclox was the most sensitive antibiotic . The average follow up was 4.9 years and the incidence of chronic osteomyelitis was 1.75% . In conclusion this study confirms that clinically it is not possible to detect a subperiosteal abscess within the first 48 hours . As blood culture is not 100% reliable in isolating the ineffective organism and incision and drainage plays a double role, firstly in the identification of the offending organism and secondly in the evacuation of pus . A six-week antibiotic therapy is essential to prevent any relapses . The correct management of acute osteomyelitis in children is surgical drainage of the affected site combined with antibiotic therapy. Biull Eksp Biol Med, 1992 Oct, 114(10), 385 - 7 {The effect of recombinant interleukin-2 on the course of experimental staphylococcal peritonitis in mice}; Shadrin OV et al.; The effect of RIL-2 on the survival of mice with S . aureus--induced peritonitis was studied . Animals received bacterial suspension and RIL-2 as following: bacteria--on days 0, +2, RIL-2--day 0 (group 1); bacteria--days 0, +4, RIL-2--days 0, +2 (group 2); bacteria--days 0, +6, RIL-2--days 0, +2, +4 (group 3) . RIL-2 exerted no protective effect in group 1 . However, in groups 2 and 3, where the control animals survival was, resp., 56% and 38%, the RIL-2 treatment increased survival up to, resp., 84% and 70% . Antibiotics given instead of RIL-2 in analogous regimen decreased the survival in group 3 to the level of 25% . Thus, RIL-2 proved to be a potent therapeutic agent in the 2nd of 3d studied models of S . aureus--induced peritonitis in mice . The perspectives of RIL-2 use in the treatment of bacterial peritonitis, including porous ones, and of the immunodepression--aggravated conditions are discussed. Eur J Haematol, 1992 Oct, 49(4), 199 - 207 Granulocyte-macrophage colony-stimulating factor (GM-CSF) and granulocyte colony-stimulating factor (G-CSF) secretion by adherent monocytes measured by quantitative immunoassays; Sallerfors B et al.; The kinetics of GM-CSF and G-CSF secretion by purified adherent human monocytes were studied by quantitative immunoassays . Interleukin-1 (IL-1); 4-40 ng/ml and E . coli lipopolysaccharide (LPS); 0.1-1.00 ng/ml, were the most effective stimuli and induced dose-dependent secretion of both GM-CSF and G-CSF . Secretion of newly synthesized CSF was detectable 3-6 hours after stimulation and continued for approximately 24 h . Twenty minutes pulse exposure to LPS was sufficient to induce half maximum secretion of GM-CSF, and after 24-36 h the adherent monocytes could not be restimulated . Neither GM-CSF nor TNF could down-regulate the secretion of GM-CSF . IL-3 induced a minor secretion of GM-CSF whereas TNF, G-CSF, M-CSF and IFN-gamma were unable to induce GM-CSF secretion . In addition to LPS and IL-1, GM-CSF and to a minor degree TNF induced G-CSF secretion . Enriched T lymphocytes secreted GM-CSF, but not G-CSF, after stimulation with PHA or staphylococcal enterotoxin A (SEA), whereas LPS and IL-1 were without stimulatory effects . We also noted that enriched T lymphocytes added to LPS-stimulated adherent monocytes at ratios of 1:10 or more inhibited, in a dose-dependent fashion, GM-CSF secretion by 13-55% . These findings add new quantitative data on CSF secretion by human monocytes. Cell, 1992 Sep 4, 70(5), 741 - 50 pp59fyn mutant mice display differential signaling in thymocytes and peripheral T cells; Stein PL et al.; We have generated mutant mice that do not express pp59fyn, a nonreceptor protein tyrosine kinase related to pp60src, by homologous recombination in embryonic stem cells . fyn- mice did not display an overt phenotype . Because fyn is associated with the T cell receptor (TCR), thymocyte and T cell signaling was analyzed in the mutant background . Cross-linking of TCR-CD3 in thymocytes led to markedly reduced calcium fluxes and abrogated proliferation, whereas mature splenic T cells retained largely normal proliferation despite depressed calcium movements and IL-2 production . Similarly, proliferation induced by Thy-1 cross-linking was reduced in thymocytes but not in splenic T cells . fyn- thymocytes were impaired at a late stage of maturation and showed limited clonal deletion to the Mls-1a self-super-antigen but not to staphylococcal enterotoxin A . These results implicate fyn as a critical component in TCR signaling in thymocytes and, potentially, in the process that determines T cell repertoire in the adult mouse. Nature, 1992 Sep 3, 359(6390), 79 - 82 Infection breaks T-cell tolerance; Rocken M et al.; Clonal deletion or clonal anergy establish tolerance in T cells that bear potentially autoreactive antigen receptors . Here we report that concomitant infection with the nematode Nippostrongylus brasiliensis breaks an established T-cell tolerance induced by injection of mice with Staphylococcus enterotoxin B (SEB) . CD4+ T cells from SEB-tolerant mice did not produce either interleukin-2 or interleukin-4 when challenged in vitro with SEB . N . brasiliensis infection of SEB-primed animals resulted in a normal expansion of SEB-tolerant CD4+V beta 8+ T cells in vivo as well as an equivalent increase of SEB-reactive, interleukin-4-producing CD4+V beta 8+ T cells both in SEB-tolerant and in normal animals . Thus, infection with N . brasiliensis circumvented the tolerance established with SEB . Activation of anergic, potentially autoreactive CD4+ T cells by infectious agents seems to be a major pathway for the initiation of autoimmune diseases . Our results suggest that infectious agents may break tolerance in potentially autoreactive CD4+ T cells by activation of alternative reaction pathways. J Med Microbiol, 1992 Sep, 37(3), 201 - 5 Purification and characterisation of elastase from Staphylococcus epidermidis; Sloot N et al.; An elastase of Staphylococcus epidermidis was purified by ion exchange chromatography on CM-Sepharose and characterised . Its M(r) is c . 21 kDa, its optimal temperature for activity is 42 degrees C and the pH optimum is 6.8 . The enzyme is activated by cysteine and other SH-donators and inhibited by L-trans-epoxy-succinylleucylamido-(4-guanidino)butane (E64), an inhibitor of cysteine proteases, but not by 3,4-dichloroisocoumarin (3,4-DCI), an inhibitor of serine proteases . This finding suggests that the elastase of S . epidermidis is a cysteine protease . Because S . epidermidis elastase degrades human sIgA, IgM, serum albumin, fibrinogen, and fibronectin, this enzyme may be regarded as a virulence factor. J Exp Med, 1992 Sep 1, 176(3), 845 - 53 Studies on T cell maturation on defined thymic stromal cell populations in vitro; Jenkinson EJ et al.; We describe an in vitro system in which positive selection of developing T cells takes place on defined stromal cell preparations, which include major histocompatibility complex class II+ epithelial cells but exclude cells of bone marrow origin . In this system, maturation of double-positive T cell receptor negative (TCR-), CD4+8+ thymocytes into single-positive TCR+, CD4+ and CD8+ cells takes place together with the development of functional competence . As in vivo, this maturation is associated with the upregulation of TCR levels as cells progress from double-positive to single-positive status . We also show that class II+ epithelial cells in these cultures are less efficient than dendritic cells in mediating the deletion (negative selection) of V beta 8+ cells by the superantigen staphylococcal enterotoxin B . For the first time, this approach provides a model in which the cellular interactions involved in both positive and negative selection can be studied under controlled in vitro conditions. Infect Immun, 1992 Sep, 60(9), 3489 - 96 Staphylococcal alpha toxin: a study with chronically instrumented awake sheep; Harshman S et al.; The in vivo responses to staphylococcal alpha toxin are reported for 15 chronically instrumented awake yearling sheep . The data obtained from a total of 30 experiments are grouped into four categories of response: no response, noted in seven experiments done on 5 sheep; pressor response, obtained seven times in 4 sheep; fluid and solute exchange, noted on six occasions in 3 sheep; and acute heart failure and death, which occurred in 10 of the 15 sheep . "No response" denoted no change in any of the measured outcome variables . The group of sheep labeled as showing "pressor response" responded to alpha toxin infusion with an increase in pulmonary artery pressure, unaccompanied by changes either in lung lymph flow or in lung mechanics . "Changes in lung fluid and solute exchange" involve increases in lung lymph flow . The harbinger of the last category, acute left heart failure leading to death, was a marked elevation in left atrial pressure . The threshold response dose in sheep is approximately 21 micrograms/kg . A very steep dose-response curve is observed, with only a narrow window of doses, 15 to 25 micrograms/kg, between the group showing no response and the group showing death from acute heart failure . The data obtained in these studies indicate that the lethal effects of alpha toxin in sheep include acute heart failure, which may be due to direct toxicity to heart muscle and/or the coronary vasculature endothelium. J Intraven Nurs, 1992 Sep-Dec, 15(5), 283 - 7 Antibiotic lock technique; Cowan CE; Central venous catheters deliver life-prolonging medications to Group IVC AIDS patients . Unfortunately, the access device frequently provides a foci for debilitating infections . A Group IVC AIDS patient experienced repeated methicillin-resistant Staphylococcus epidermidis colonization and infection of his central venous access device . An antibiotic lock technique was created using a flush solution of vancomycin 1 mg/ml with 100 unit heparin/ml . The patient locked his catheter with 2.0 ml every evening for a period of 5 days . Repeat catheter and peripheral culturing occurred 72 hours after the 5-day period . After using the antibiotic lock technique, the patient maintained his catheter without further infection for an additional 2 months . This new approach may represent improved therapy and prevent the hazards of catheter exchange and systemic antibiotic therapy. Zentralbl Veterinarmed B, 1992 Sep, 39(7), 519 - 25 Isolation and characterization of immunoglobulin binding proteins from Staphylococcus intermedius and Staphylococcus hyicus; Greene RT et al.; 125-I-IgG binding activities were observed with 15 (17%) of 90 S . intermedius isolates from dogs and 39 (95%) of 41 S . hyicus isolates from pigs . Binding activities were not detected with S . hyicus isolates from cows . The IgG binding proteins of 2 S . intermedius, 2 S . hyicus, and protein A from S . aureus Cowan I were isolated from their cell surfaces . The proteins precipitated with IgG preparations from human, rabbit, pig, dog and horse, but not with IgG from cow, mouse and chicken . This indicated that these IgG binding proteins could be classified as type I receptors . In addition, the isolated proteins from all 3 staphylococcal species precipitated with polyclonal chicken anti-protein A antiserum . SDS-PAGE, Western blotting and gel isoelectric focussing of the proteins revealed numerous bands in the 42,000 D range and acid isoelectric points . The isoelectric point of the isolated proteins from both S . intermedius cultures was slightly more acidic than those from S . hyicus and S . aureus . The present results indicate a close functional and antigenic similarity, if not identity, between IgG binding proteins of S . intermedius and S . hyicus, and protein A of S . aureus. Antibiot Khimioter, 1992 Sep, 37(9), 25 - 7 {Augmentin in the clinical picture of infectious diseases}; Gracheva NM et al.; Augmentin is a new combination manufactured by Smith Kline Beecham (Great Britain) . It is composed of amoxycillin and clavulanic acid and has antibacterial activity . Augmentin was used in clinical trials in the therapy of 50 adult patients with suppurations after surgical operations on the organs of the abdominal cavity, general staphylococcal infections, pneumonia and prophylactically during the preoperative period . It was also used in the treatment of 30 children patients with bronchopulmonary affections and inflammatory otorhinolaryngological diseases . The clinical trials were performed in the Clinic of Infectious Diseases of the N . G . Gabrichevskii Moscow Research Institute of Epidemiology and Microbiology . For comparison ampicillin was used in the trials . Augmentin was shown to be an efficient formulation with antibacterial activity which could be successfully used in the parenteral therapy of severe affections due to organisms sensitive to it . In the treatment of the children patients with pneumonia augmentin by its therapeutic efficacy proved to be superior to ampicillin . The tolerance of augmentin was good. Ann Pediatr (Paris), 1992 Sep, 39(7), 397 - 406 {Pustular dermatoses in the neonatal period}; Moisson YF et al.; The diagnosis of pustular dermatosis occurring during the first months of life is usually based on clinical findings . However, some cases may require a few simple investigations including cytological studies, cultures, and skin biopsies . The most common causes of infectious pustular skin lesions include bacterial infections, which may be septicemic (with Listeria as the leading causative agent) or initially localized (staphylococcus); viral infections (varicella, herpes); fungal infections, i.e., candidiasis (congenital or neonatal) or the very recently described form of pustulosis due to Malassezia furfur; or parasitic (scabies) . The main benign transient neonatal forms of pustulosis include infantile acropustulosis (for which the relationship with scabies is discussed), toxi-allergic erythema, transient pustular melanosis, and neonatal acne . Lastly, rare causes of neonatal pustulosis are reviewed . The need for investigating every neonate with skin pustules for an infectious disease, especially due to a bacterium, is emphasized. J Clin Microbiol, 1992 Sep, 30(9), 2385 - 90 Phenotypic variation of Staphylococcus epidermidis isolated from a patient with native valve endocarditis; Deighton M et al.; Two colonial variants of Staphylococcus epidermidis were isolated from the valvular tissue of a patient with native valve endocarditis . In addition to differing in colonial morphology, the two variants differed in hemolysis on blood-containing media, in adherence capacity, and in the expression of certain enzymes . Under suitable conditions, both variants were themselves capable of phenotypic variation, although they differed in the rate at which variants were generated . The variants yielded identical profiles on restriction endonuclease analysis of plasmid DNA and pulsed-field gel electrophoresis of whole-cell DNA . This report suggests a possible role for phenotypic variation in coagulase-negative staphylococcal virulence . Congo red agar would be an excellent medium for studying the contribution of variation to the virulence of these organisms. FEMS Microbiol Immunol, 1992 Sep, 5(1-3), 93 - 100 A simple method for the determination of the pore radius of ion channels in planar lipid bilayer membranes; Krasilnikov OV et al.; A new method of pore size determination is presented . The results of applying this simple method to ion channels formed by staphylococcal alpha-toxin and its N-terminal fragment as well as to cholera toxin channels are shown . The advantages and the difficulties of this method are discussed . It was found that (i) the mobility of ions in solutions depends only on the percentage of concentration of added non-electrolytes and practically not on their chemical nature (sugars or polyglycols) and molecular size; (ii) the proportional change of both ion channel conductance and bulk solution conductivity by low M . nonelectrolytes may be used as an indication of a diffusion mechanism of ion transport through channels; (iii) the slope of the dependence of the ion channel conductance on the bulk conductivity of solutions containing different concentrations of non-electrolyte is a good measure of channel permeability for non-electrolytes. Ann Neurol, 1992 Sep, 32(3), 352 - 7 Bacterial toxin superantigens activate human T lymphocytes reactive with myelin autoantigens; Burns J et al.; Some bacteria that are common human pathogens produce protein toxins that are potent activators of human T lymphocytes expressing certain types of T-cell receptors . In this study we examined the ability of staphylococcal toxins to stimulate human T lymphocytes that also recognized the myelin autoantigens myelin basic protein and proteolipid protein . T-cell populations responding to myelin basic protein or proteolipid protein were isolated from 4 subjects including 1 individual with multiple sclerosis . All myelin antigen-specific T cells responded in proliferation studies to at least one of the nine superantigenic toxins used in this study . The superantigenic toxins were up to 7 x 10(5)-fold more potent in proliferation assays than the myelin antigens to which the T cells were initially sensitized . In addition, cytotoxic, myelin basic protein-reactive T lymphocytes lysed antigen-presenting cells incubated with superantigenic toxins . These findings demonstrate a mechanism by which some bacterial infections might produce activation of myelin basic protein- and proteolipid protein-reactive T lymphocytes and perhaps contribute to demyelinating disease in humans. J Immunol, 1992 Sep 1, 149(5), 1497 - 503 Superantigen-mediated human monocyte-T lymphocyte interactions are associated with an MHC class II-, TCR/CD3-, and CD4-dependent mobilization of calcium in monocytes; Damaj B et al.; The present study was designed to examine the potential involvement of calcium ions as second messengers in the mediation of the staphylococcal enterotoxin A (SEA)/MHC class II-induced activation of human monocytes . Treatment of monocytes with a monomeric form of SEA failed to induce detectable changes in the level of intracellular calcium in either monocytes or THP-1 cells . However, cross-linking of SEA with biotin-avidin induced a rapid and transient increase in calcium levels in monocytes and in INF-gamma-treated THP-1 cells . This artificial cross-linking system was reproduced by natural physiologic ligands expressed on the surface of T lymphocytes . Delayed, transient, and concentration (cell as well as toxin)-dependent increases in the cytoplasmic level of free calcium in SEA-treated monocytes were observed upon the addition of autologous resting T cells or purified CD4+ cells, but not of CD8+ cells, B cells, or neutrophils . Antibodies against MHC class II Ag, TCR/CD3, and CD4 molecules inhibited the SEA-dependent interaction between monocytes and T cells as indicated by significant decreases in the rise of calcium levels observed in monocytes . Anti-CD8 and anti-class I antibodies did not affect the interaction between the monocytes and the T cells and failed to alter the calcium response . Taken together, these results suggest that the SEA-induced, T cell-dependent calcium mobilization in monocytes requires physical interactions between SEA-MHC class II, TCR/CD3, and CD4 molecules . The ability to mediate a T cell-dependent calcium increase in monocytes was shared by several enterotoxins including staphylococcal enterotoxin B and toxic shock syndrome toxin-1 . The characteristics of the SEA-mediated calcium mobilization in monocytes strongly support the hypothesis that this response is an integral part of the signal transducing machinery linked to MHC class II molecules. Appl Microbiol Biotechnol, 1992 Sep, 37(6), 772 - 83 Enzymatic preparation of 1,6-anhydro-muropeptides by immobilized murein hydrolases from Escherichia coli fused to staphylococcal protein A; Engel H et al.; In order to produce biologically active 1,6-anhydro-muropeptides in large amounts by enzymatic degradation of isolated bacterial murein polymer highly specific periplasmic murein-metabolizing enzymes from Escherichia coli are made available . The genes slt, dacB, and mepA, encoding the soluble lytic transglycosylase (Slt), the penicillin-sensitive DD-endopeptidase (PBP4), and the penicillin-insensitive murein endopeptidase A (MepA), were independently fused to the N-terminal encoding sequence of staphylococcal protein A (SpA) under control of the temperature-inducible phage lambda pR promoter . The SpA fusion proteins were stably over-produced at high levels in E . coli upon temperature induction at 42 degrees C and account for 3% (5 mg SpASlt/l culture), 3% (5 mg SpAPBP4/l culture), and 0.3% (0.5 mg SpAMepA/l culture) of total protein . The SpA fusion proteins, immobilized on IgG Sepharose, are proteolytically sensitive, in vitro, resulting in complete degradation of the SpA portion of the fusion proteins and release of the murein hydrolases in intact and enzymatically active form into the supernatant . Proteolytic degradation could be prevented by p-hydroxymercuribenzoic acid (PHMB) or ethylenediaminetetraacetate (EDTA) suggesting the involvement of the periplasmic protease Pi from E . coli . The immobilized fusion proteins were enzymatically active and could be used for the batch production of biologically active 1,6-anhydro-muropeptides, which were successively separated on HPLC . Isolated murein polymer was degraded quantitatively to monomeric 1,6-anhydro-muropeptides when immunoglobulin G (IgG)-SpASlt was used in combination with IgG-SpAMepA . A combination of IgG-SpASlt with IgG-SpAPBP4 left the 1,6-anhydro-dimers and oligomers being cross-linked via an LD-peptide bond (m-DAP-m-DAP) uncleaved. J Biotechnol, 1992 Sep, 25(3), 269 - 87 Fusions to the 5' end of a gene encoding a two-domain analogue of staphylococcal protein A; Rondahl H et al.; A novel gene fusion system has been constructed for fusions to the 5' end of gene zz, encoding a two-domain analogue of staphylococcal protein A designated ZZ . Four different genes were fused to the 5' end of zz, and their gene products were analyzed . One of the genes encodes a protein located intracellularly in Escherichia coli and the other three genes encode gene products destined for secretion across the cytoplasmic membrane by the presence of an amino terminal signal sequence . After production in E . coli, the fusion proteins were purified in a single step by IgG-affinity chromatography . The purified ZZ fusions could be used directly for amino terminal sequencing to confirm the start of translation of the intracellular product and the processing of the signal peptide of the translocated products . This is the first example of ZZ fusions to the C-terminus of gene products . To simplify the general use of fusions to the 5' end of zz, a new plasmid vector was constructed containing a multi restriction enzyme cloning linker and the lacZ' gene which enables screening for production in alpha-complementing supE strains of E . coli on indicator plates. J Hosp Infect, 1992 Sep, 22(1), 33 - 9 Plasmid-pattern analysis in the differentiation of Staphylococcus epidermidis isolates; Ferreiros CM et al.; Plasmids carried by 29 Staphylococcus epidermidis isolates from two different origins (invasive and carrier, non-hospital-associated strains) were analysed by agarose gel electrophoresis in order to evaluate any association with virulence . Plasmid patterns were found to be characteristic of individual strains and no particular molecular size band was found to correlate with either of the two categories of isolates . It is concluded that the plasmid pattern analysis carried out could not differentiate between carrier and invasive S . epidermidis, but did provide a useful tool for epidemiological investigation. Proc Natl Acad Sci U S A, 1992 Sep 1, 89(17), 8035 - 9 Superantigen staphylococcal enterotoxin B-induced T-helper cell activation is independent of CD4 molecules and phosphatidylinositol hydrolysis; Oyaizu N et al.; The role of the CD4 molecule in activation of T-helper cells was examined by investigating the effect of an anti-CD4 monoclonal antibody (Leu3a) in conventional peptide antigen-specific cloned T-helper cells that are also reactive to staphylococcal enterotoxin B (SEB) . These T-helper cell clones are CD4+/CD45RO+/T-cell antigen receptor beta-chain variable region 12-positive and can respond to nominal peptide antigens and SEB by proliferation in the presence of class II major histocompatibility complex-expressing accessory cells . Although antigen and SEB were comparable in their ability to induce proliferative responses, interleukin 2 (IL-2) production, and IL-2 receptor alpha-chain expression, stimulation with SEB failed to trigger phosphatidylinositol hydrolysis or a rise in the intracellular free calcium ion concentration . Leu3a treatment inhibited antigen-induced proliferative responses of T cells with concomitant suppression of IL-2 production and IL-2 receptor expression . In contrast, SEB-induced responses were unaffected by Leu3a . These findings indicate that the functional consequences of binding (ligation) of conventional antigen and of superantigen with the T-cell receptor are distinct in the context of both signal transduction pathways and participation of CD4 molecules. Scand J Immunol, 1992 Sep, 36(3), 479 - 86 Staphylococcus albus-induced protein kinase C translocation in human neutrophils: the effect of opsonization, cytochalasin B, pertussis toxin, intra- and extracellular calcium, and R59022; Obel N; Membrane-associated protein kinase C has been proposed to be essential in transmembrane signalling systems activating neutrophils . A main function of the neutrophil is phagocytosis and killing of microorganisms . Nevertheless, previously published reports mainly have described the effect of artificial or soluble stimulators upon neutrophil protein kinase C activity . Therefore, membrane-associated protein kinase C was studied in neutrophils stimulated by Staphylococcus albus . The bacteria were found to induce a striking increase in membrane-associated protein kinase C, an effect which depended upon a previous opsonization of the bacteria . Cytochalasin B, which inhibits phagocytosis, was shown to abrogate S . albus-induced protein kinase C translocation . Chelation of intracellular calcium totally abolished S . albus-induced protein kinase C translocation, a phenomenon that could not exclusively be ascribed to chelation of extracellular calcium . The diacylglycerol kinase inhibitor R59022, which has been reported to increase endogenous diacylglycerol accumulation, nearly doubled the effect of S . albus upon membrane-associated protein kinase C . Pertussis toxin in concentrations which completely inhibited fLMP-induced superoxide generation did not affect S . albus-induced protein kinase C translocation . It is concluded that phagocytosis of S . albus is accompanied by a translocation of protein kinase C to the cell membrane, a phenomenon that relies upon enhanced diacylglycerol production and calcium transients and occurs independently of pertussis toxin-inhibitable G-proteins. Ginekol Pol, 1992 Sep, 63(9), 447 - 50 {Evaluating the usefulness of qualitative analysis of latex c-reactive protein for diagnosing neonatal sepsis}; Grzywna W et al.; We analyzed the results of determinations C-reactive protein (Latex Test Dialab firm) of 136 newborns . We divided them into three groups: with generalized infections, with infection without sepsis and without infection . Sensitivity of method for neonates with septicaemia was 60%, for neonates with infection without sepsis--81.25%, together for all infections--66.1% . After overlooking sepsis caused by Staphylococcus epidermidis sensitivity for sepsis was 81.8% and for all infections--81.6%. Can J Microbiol, 1992 Sep, 38(9), 937 - 44 Receptors for toxic shock syndrome toxin-1 and staphylococcal enterotoxin A on human blood monocytes; See RH et al.; Staphylococcal toxic shock syndrome toxin-1 (TSST-1) as well as staphylococcal enterotoxin A (SEA) and B (SEB) have recently been shown to bind directly to the class II major histocompatibility antigen, HLA-DR . Whereas others have characterized TSST-1 and SEA binding to HLA-DR on transfected L cells or B lymphoma cell lines, we sought evidence for direct binding of TSST-1 and SEA to HLA-DR on purified human monocytes . A single class of high-affinity receptors was found for both TSST-1 (dissociation constant (Kd) 40 nM, 3.4 x 10(4) receptors per cell) and SEA (Kd 12 nM, 3.2 x 10(4) receptors per cell) on normal human monocytes . Affinity cross-linking of 125I-labeled toxins to monocytes revealed the presence of two membrane protein subunits with molecular masses consistent with the alpha and beta chains of human HLA-DR (35 and 28 kDa, respectively) . The anti-HLA-DR monoclonal antibody L243, but not L203 or 2.06, inhibited radiolabeled toxin binding to human monocytes and neutralized the mitogenic response of human T lymphocytes to both toxins . However, L243 was consistently more effective in blocking radiolabeled TSST-1 than SEA binding to human monocytes from the same donors, suggesting that TSST-1 and SEA may be binding to overlapping epitopes rather than to the same epitope on HLA-DR . Because TSST-1 and SEB bind to distinct epitopes on HLA-DR and because SEA cross competes with both TSST-1 and SEB on the HLA-DR receptor, we postulate that SEA occupies a binding site within HLA-DR that overlaps both TSST-1 and SEB.(ABSTRACT TRUNCATED AT 250 WORDS) Pol Tyg Lek, 1992 Aug 18-31, 47(34-35), 735 - 8 {IgE-containing immune complexes in bronchial asthma}; Jarzab J et al.; Level of circulating immunological complexes and their immunoglobulin content have been determined in 36 asthmatic patients, including 15 patients with atopic asthma and 21 patients with infectious asthma . A technique of staphylococcal protein A binding has shown, that the level of the circulating immunological complexes is increased in patients with infectious bronchial asthma . An amount of IgE in these complexes has been increased in both atopic and infectious bronchial asthma . However, a level of IgE-containing immunological complexes has been higher in the atopic asthma, then that in infectious form of the disease . An increased IgA content in the immunological complexes has been noted in the infectious asthma. Proc Natl Acad Sci U S A, 1992 Aug 15, 89(16), 7727 - 31 T-cell antigen receptor binding sites for the microbial superantigen staphylococcal enterotoxin A; Pontzer CH et al.; We have examined the interaction of the microbial superantigen staphylococcal enterotoxin A (SEA) with peptides corresponding to overlapping regions of the T-cell antigen receptor beta chain variable region V beta 3 . SEA is known to stimulate murine T cells bearing certain V beta elements, among them V beta 3 . Five peptides were synthesized representing amino acids 1-24, 20-44, 39-60, 57-77, and 74-95 of V beta 3 . We demonstrate here that soluble V beta 3-bearing beta chains can bind to a complex of SEA and major histocompatibility complex class II and that the synthetic peptide V beta 3-(57-77) blocked this interaction . The peptide V beta 3-(57-77) also inhibited SEA-induced interferon-gamma production and SEA-induced proliferation of B10.BR spleen cells . Conversely, the peptide corresponding to amino acids 57-77 of V beta 8.2, a V beta element that is not recognized by SEA, decreased staphylococcal enterotoxin C-2-induced proliferation but did not affect SEA-induced proliferation . The peptide inhibition of SEA-induced function was due at least in part to inhibition of V beta 3-bearing T-cell activity, since the percentage of T cells reactive with an anti-V beta 3 monoclonal antibody was significantly reduced by V beta 3-(57-77) . These data suggest that the region of V beta 3 encompassing amino acids 57-77 is an area that displays the appropriate sequence and conformation for binding of the SEA molecule and blocking of the resultant interaction with the T-cell antigen receptor. J Immunol, 1992 Aug 15, 149(4), 1156 - 63 In vivo induction of apoptosis in immature thymocytes by staphylococcal enterotoxin B; Lin YS et al.; Staphylococcal enterotoxins are potent T cell mitogens . Recent studies have shown that the binding of these toxins to class II MHC molecules on accessory cells is essential for the stimulation of T cells which bear specific V beta segment of TCR . In the present study we show that i.v . administration of staphylococcal enterotoxin B (SEB) results in an enlargement of spleen and lymph nodes but causes thymus atrophy . Elimination of CD4+CD8+ cells predominantly accounted for the shrinkage of thymus, and the lowest level of this cell population was reached 4 days after SEB injection . Furthermore, this decrease in CD4+CD8+ cells was accompanied by a relative increase in the percentages of CD4+CD8-, CD4-CD8+ and CD4-CD8- cells, whereas their absolute numbers actually reduced on day 4 . The thymus shrinkage involved apoptosis which was characterized by DNA fragmentation and morphologic changes . The depletion of Thy-1 high, TCR-alpha beta low and TCR-alpha beta intermediate cells also occurred with a kinetic correlated to the reduction of CD4+CD8+ cells . Our results further showed that the percentages of V beta 8+ cells reduced 12 h post SEB injection, increased after 2 days, and decreased again thereafter . SEB thus causes both apoptotic and stimulative effects in the thymus . Apparently, the tremendous loss of double-positive cells (greater than 90% in cell number on day 4) is not simply due to the reduction of V beta 8+ cells, the possible modulatory effect of other factors or hormones which may play a role in the cell death is discussed. J Bacteriol, 1992 Aug, 174(16), 5354 - 61 Purification and characterization of EpiD, a flavoprotein involved in the biosynthesis of the lantibiotic epidermin; Kupke T et al.; The plasmid-encoded epidermin biosynthesis gene, epiD, of Staphylococcus epidermidis Tu3298 was expressed in Escherichia coli by using both the malE fusion system and the T7 RNA polymerase-promoter system . EpiD was identified by Western blotting (immunoblotting) with anti-maltose-binding protein (MBP)-EpiD antiserum . EpiD and the MBP-EpiD fusion protein, which were mainly present in the soluble protein fraction, were purified from the respective E . coli clones . Purified EpiD showed the typical absorption spectrum of an oxidized flavoprotein with maxima at 274, 382, and 453 nm . The coenzyme released from EpiD by heat treatment was identified as flavin mononucleotide . S . epidermidis Tu3298/EMS11, containing a mutation within epiD, was unable to synthesize active epidermin . This mutated gene, epiD*, was cloned in E . coli and expressed as an MBP-EpiD* fusion protein . DNA sequencing of epiD* identified a point mutation that led to replacement of Gly-93 with Asp . Unlike MBP-EpiD, the fusion protein MBP-EpiD* could not bind flavin mononucleotide . We propose that EpiD catalyzes the removal of two reducing equivalents from the cysteine residue of the C-terminal meso-lanthionine to form a --C==C-- double bond and is therefore involved in formation of the unusual S-{(Z)-2-aminovinyl{-D-cysteine structure in epidermin. Arch Surg, 1992 Aug, 127(8), 893 - 7; discussion 897-8 Postinjury shock and early bacteremia . A lethal combination; Moore FA et al.; Gut bacteria translocation has been invoked as a prime cause of early postinjury death . To examine this hypothesis, we obtained emergency department blood cultures in 132 acutely injured patients requiring urgent laparotomy for trauma . In the latter half of these patients, mesenteric lymph node and liver biopsy cultures were also performed . The incidence of early bacteremia was 11% (10/94) in the patients without shock compared with 32% (12/38) in the group with shock . The majority (73%) were gram-positive bacteremias . Most notably, Staphylococcus was isolated in 13% (5/38) of the patients with shock, but these isolates were of no apparent clinical significance . In contrast, 18% (7/38) of the patients with shock had enteric bacteremias, and all of these patients died . Cultures were positive in 11% of the liver samples and 15% of the mesenteric lymph nodes . With the exception of two patients with concurrent enteric bacteremias, these hepatic and mesenteric lymph node bacteria were of no clinical significance . In conclusion, bacterial translocation occurs infrequently, and virtually all enteric bacteria were found in dying patients; the cause or effect remains to be defined. Invest Ophthalmol Vis Sci, 1992 Aug, 33(9), 2650 - 63 Immune response to Staphylococcus epidermidis-induced endophthalmitis in a rabbit model; Pleyer U et al.; Although Staphylococcus epidermidis is the most common cause of postoperative pseudophakic endophthalmitis, little is known about the immune response to S . epidermidis-induced endophthalmitis . Using a rabbit model, the immune response to an intravitreal injection of 7000 S . epidermidis (group 1) or 30,000 S . epidermidis (group 2) organisms was investigated . Clinical evaluations showed that rabbits in group 2 had a more severe inflammatory reaction in the conjunctiva, cornea, iris, and vitreous than those in group 1 . The inflammatory reaction in group 1 largely resolved by day 30; group 2 continued to show a severe inflammatory response . Histopathologic findings correlated with clinical findings, with rabbits in group 2 showing a more severe inflammatory reaction in both the anterior and posterior segments of the globe . Positive vitreous cultures for S . epidermidis were present in rabbits in group 1 on days 3, 7, 10, 14, and 21 but not thereafter . However, group 2 had higher vitreous colony counts at days 3, 7, and 14 and negative vitreous cultures thereafter . Neither group showed delayed hypersensitivity to S . epidermidis antigens (evaluated by skin tests) . Serum immunoglobulin (Ig) G antibody levels to phenol-inactivated S . epidermidis and glycerol teichoic acid (GTA) increased progressively, reached a peak at days 10-14, and then declined in both groups . Serum IgA antibody levels to these antigens were not detected . Group 2 had a more prolonged IgG antibody response in vitreous and aqueous than group 1 . Tear fluid showed the weakest IgG and IgA antibody response to S . epidermidis and GTA . S . epidermidis-induced endophthalmitis was associated with a humoral but not a delayed hypersensitivity response to this organism. Infect Immun, 1992 Aug, 60(8), 3456 - 9 Staphylococcal toxic shock syndrome toxin 1-induced tumor necrosis factor alpha and interleukin-1 beta secretion by human peripheral blood monocytes and T lymphocytes is differentially suppressed by protein kinase inhibitors; See RH et al.; The signal transduction pathways by which staphylococcal toxic shock syndrome toxin 1 (TSST-1) induces tumor necrosis factor alpha (TNF-alpha) and interleukin-1 beta (IL-1 beta) secretion were examined with various protein kinase inhibitors . TNF-alpha secretion by normal human monocytes and T cells in response to TSST-1 was suppressed by inhibitors of protein kinase C (H7) and tyrosine kinases (genistein) . In contrast, the secretion of IL-1 beta was blocked by a cyclic AMP- and cyclic GMP-dependent kinase inhibitor (HA1004) as well as by H7 and genistein . These results suggest that the secretion of TNF-alpha and IL-1 beta may be differentially regulated by TSST-1 and that protein kinases play an important role in mediating cytokine responses to the toxin. Proteins, 1992 Aug, 13(4), 275 - 87 NMR docking of a substrate into the X-ray structure of staphylococcal nuclease; Weber DJ et al.; The conformation of the staphylococcal nuclease-bound metal-dTdA complex, previously determined by NMR methods {Weber, D.J., Mullen, G.P., Mildvan, A.S . (1991) Biochemistry 30:7425-7437} was docked into the X-ray structure of the enzyme-Ca(2+)-3',5'-pdTp complex {Loll, P.J., Lattman, E.E . (1989) Proteins: Struct., Funct., Genet . 5:183-201} by superimposing the metal ions, taking into account intermolecular nuclear Overhauser effects from assigned aromatic proton resonances of Tyr-85, Tyr-113, and Tyr-115 to proton resonances of the leaving dA moiety of dTdA, and energy minimization to relieve small overlaps . The proton resonances of the Phe, Tyr, and Trp residues of the enzyme in the ternary enzyme-La(3+)-dTdA complex were sequence specifically assigned by 2D phase-sensitive NOESY, with and without deuteration of the aromatic protons of the Tyr residues, and by 2D heteronuclear multiple quantum correlation (HMQC) spectroscopy and 3D NOESY-HMQC spectroscopy with 15N labeling . While resonances of most Phe, Tyr and Trp residues were unshifted by the substrate dTdA from those found in the enzyme-La(3+)-3',5'-pdTp complex and the enzyme-Ca(2+)-3',5'-pdTp complex, proton resonances of Tyr-85, Tyr-113, Tyr-115, and Phe-34 were shifted by 0.08 to 0.33 ppm and the 15N resonance of Tyr-113 was shifted by 2.1 ppm by the presence of substrate . The optimized position of enzyme-bound dTdA shows the 5'-dA leaving group to partially overlap the inhibitor, 3',5'-pdTp (in the X-ray structure) . The 3'-TMP moiety of dTdA points toward the solvent in a channel defined by Ile-18, Asp-19, Thr-22, Lys-45, and His-46 . The phosphate of dTdA is coordinated by the metal, and an adjacent inner sphere water ligand is positioned to donate a hydrogen bond to the general base Glu-43 and to attack the phosphorus with inversion . Arg-35 and Arg-87 donate monodentate hydrogen bonds to different phosphate oxygens of dTdA, with Arg-87 positioned to protonate the leaving 5'-oxygen of dA, thus clarifying the mechanism of hydrolysis . Model building of an additional 5'-dGMP onto the 3'-oxygen of dA placed this third nucleotide onto a surface cleft near residues Glu-80, Asp-83, Lys-84, and Tyr-115 with its 3'-OH group accessible to the solvent, thus defining the size of the substrate binding site as accommodating a trinucleotide. Clin Nephrol, 1992 Aug, 38(2), 86 - 9 Heparin-induced thrombocytopenia in renal failure; Hall AV et al.; A 35-year-old patient with symptomatic chronic renal failure secondary to hereditary nephritis underwent heparinization during urgent hemodialysis . Subclavian catheter insertion was complicated by staphylococcal septicemia which precluded Gore-tex graft placement . Arteriovenous fistula failed immediately postoperatively and thrombectomy was required . Heparin-induced thrombocytopenia was suspected and the test was strongly positive . Acetyl salicylic acid (ASA) 40 mg, was used with heparin for dialysis, and there were no episodes of gastrointestinal hemorrhage or access site thrombosis . The use of very low-dose ASA in this setting is reported. J Leukoc Biol, 1992 Aug, 52(2), 202 - 8 Effects of antiorthostatic suspension and corticosterone on macrophage and spleen cell function; Kopydlowski KM et al.; The purpose of this study was to determine whether antiorthostatic suspension of C3HeB/FeJ mice for a period of 11 days affected macrophage and spleen cell function . We found that antiorthostatic suspension did not alter macrophage secretion of prostaglandin E2, tumor necrosis factor alpha, and interleukin-1 . Antiorthostatic suspension also did not affect macrophage-mediated contact-dependent cytotoxicity, TNF-mediated cytotoxicity, expression of class II histocompatibility molecules, or concanavalin A and Bandeiraea simplicifolia lectin binding sites . The proliferative response of splenic T cells in response to mitogens and staphylococcal exotoxins was significantly enhanced in antiorthostatically suspended mice . We detected significantly higher concentrations of corticosterone in the plasma of antiorthostatically suspended mice . Therefore, there did not appear to be any direct immunosuppressive effects of corticosterone on the parameters tested. J Exp Med, 1992 Aug 1, 176(2), 575 - 9 Memory T cells are anergic to the superantigen staphylococcal enterotoxin B; Lee WT et al.; We have used staphylococcal enterotoxin B (SEB) to study the role of naive and memory T cells in the induction of peripheral tolerance . After administration of SEB to mice, the numbers of naive and memory T cells increase, as does the proportion of memory T cells, which are unresponsive to further stimulation with SEB in vitro . In addition, memory T cells generated in response to conventional antigen, which proliferate and provide help to B cells in the presence of the conventional antigen, fail to respond to superantigen . Hence, memory T cells, in general, are anergized by SEB . These results suggest that SEB-induced activation and anergy reflect the combined responses of naive and memory T cells . The differential activation vs . anergy of naive and memory T cells by superantigen may be related to cytokine production and may play an important role in the etiology of autoimmune diseases or immunodeficiency diseases such as acquired immune deficiency syndrome. J Clin Microbiol, 1992 Aug, 30(8), 1948 - 52 Staphylococcus lugdunensis endocarditis; Shuttleworth R et al.; Staphylococcus lugdunensis is a recently described coagulase-negative species which has been associated with human infections, including infective endocarditis . A case of native valve endocarditis caused by this organism is described . The initial laboratory detection of S . lugdunensis is facilitated by a positive test for ornithine decarboxylase . The identification of such isolates should not cause difficulty unless undue reliance is placed upon a small number of tests. Epidemiol Infect, 1992 Aug, 109(1), 59 - 68 Restriction endonuclease fingerprinting of genomic DNA of Staphylococcus species of bovine origin; Matthews KR et al.; Fifty-one staphylococcal isolates from mammary secretions of cows with subclinical mastitis were examined by antibiograms and DNA restriction endonuclease fingerprinting (REF) . DNA REF differentiated closely related strains of each species isolated from mammary secretions of different mammary glands of the same cow and from the same mammary gland at different periods of the lactation cycle . In addition, REF analysis provided evidence concerning persistence of infection in the same or different mammary gland over different periods of the lactation cycle, and occurrence of infection with similar and dissimilar strains of each Staphylococcus species . Antibiograms were of limited value in differentiating closely related strains . The ease by which REF analysis can be performed together with the reproducibility and clarity of REF patterns suggest that this technique is useful for differentiating closely related and unrelated strains of Staphylococcus species isolated from bovine mammary secretions. Burns, 1992 Aug, 18(4), 332 - 5 Predominance of staphylococcal organisms in infections occurring in a burns intensive care unit; Taylor GD et al.; To assess the sites, incidence, and bacteriology of infections in intensive care burn patients, a prospective survey of all admissions to a tertiary care institution burn unit was carried out over a 12-month period . One hundred and sixteen patients were admitted, 106 with a diagnosis of thermal burns . Forty patients developed 90 infections . Only two deaths occurred, one in a patient with sepsis . In order of frequency, pneumonia, burn infection, UTI and primary bacteraemia were most common . Staphylococcal species accounted for a majority of infections at all body sites except UTI (47 per cent of all infections, including 11 of 14 bacteraemic infections) . Staph . aureus sepsis was more common in those carrying the organism on admission . Strain typing of paired admission and subsequent clinical isolates in 19 patients with Staph . aureus sepsis indicated that eight (42 per cent) became infected with a strain they carried on admission . Further reductions in septic complications of burns in our center would be best directed at staphylococcal species, particularly Staph . aureus . Both eradication of carrier state, and prevention of acquisition of Staph . aureus strains could be explored. J Clin Pathol, 1992 Aug, 45(8), 716 - 21 Staphylococcal toxins and sudden infant death syndrome; Malam JE et al.; AIMS: To investigate the hypothesis that commonly occurring bacterial toxins cause sudden infant death syndrome (SIDS) by (1), determining in which tissues bacterial toxins are concentrated after intravenous injection in rats; and (2), seeing if the same tissues contain detectable toxins in cases of SIDS . METHODS: The tissue distribution of intravenously injected staphylococcal enterotoxin A (SEA), enterotoxin B (SEB), enterotoxin C (SEC), enterotoxin D (SED), toxic shock syndrome toxin (TSST-1), and alpha-haemolysin was studied in rats using immunohistology and polyacrylamide gel electrophoresis with immunoblotting . Immunostaining was also carried out on formalin fixed kidneys from cases of SIDS and a comparison series of necropsy cases using anti-SEA, anti-SEB, anti-SEC2 and anti-SED . RESULTS: Immunohistology showed that SEB, SEC, SED and TSST-1 were all concentrated in the proximal convoluted tubular cells of the kidney . The presence of these toxins was confirmed in kidney homogenates using electrophoresis and immunoblotting . There was positive granular staining in the proximal convoluted tubular cells of the kidney in 36% of SIDS cases and 12% of the comparison series with anti SEC2 (chi 2 = 6; p < 0.025) . CONCLUSION: SEC, or a bacterial toxin with epitopes in common, could have a pathogenic role in SIDS. J Antimicrob Chemother, 1992 Aug, 30(2), 135 - 9 In-vitro efficacy of a central venous catheter complexed with iodine to prevent bacterial colonization; Jansen B et al.; Infections of central venous lines are still a problem in daily medicine . Despite adequate antibiotic therapy, removal of an infected catheter often becomes necessary . A simple procedure has been developed by which a special hydrophilic central venous catheter (Secalon-Hydrocath) can be loaded with iodine . Iodine is complexed in the hydrophilic polyvinylpyrrolidone surface coating of the Hydrocath catheter and is released during contact with an aqueous medium . The amount of complexed iodine depends on the incubation time in Lugol's solution . Antimicrobial activity of the loaded catheters was assessed with Staphylococcus epidermidis, showing complete inhibition of bacterial adherence to the catheters for the duration of iodine release . Depending on the experimental conditions, iodine released from the catheter is also active on bacteria in the surrounding medium. Int Immunol, 1992 Aug, 4(8), 851 - 9 Clonal unresponsiveness results from an interaction between staphylococcal enterotoxin B and T cells expressing unexpected V beta elements; Liu H et al.; The ability of staphylococcal toxins to stimulate large numbers of T cells has led to their designation as a superantigen . Previous studies have indicated that activation of T cells bearing particular V beta elements may be responsible for the toxic effects of these bacterial products . However, the widespread expression of functionally similar proteins by unrelated bacterial species suggests the possibility that these products may represent a successful microbial strategy for subversion of the host antibacterial response . We have examined the effects of the staphylococcal enterotoxin B (SEB) on T cell clones that express different V beta elements . We note that SEB stimulates clones bearing previously defined V beta elements to proliferate and to produce cytokines . In addition, we demonstrate that an interaction between SEB and the TCR of clones that express additional V beta elements, including V beta 2 and V beta 6, results in a sterile form of immunological activation . This activation phenotype is characterized by proliferation without detectable cytokine production and is followed by profound immunological unresponsiveness in vitro and in vivo . We propose that reduced levels of antibacterial responses resulting from this form of T cell unresponsiveness may account for the highly conserved expression of superantigens by diverse bacterial species. Eur J Immunol, 1992 Aug, 22(8), 2033 - 9 Presentation of superantigen by human T cell clones: a model of T-T cell interaction; Nisini R et al.; Superantigens (SAg) interact with T lymphocytes bearing particular V beta sequences as part of their T cell receptor (TcR) . The interaction, however, requires the presence of major histocompatibility complex (MHC) class II molecules on antigen-presenting cell (APC) . In peculiar circumstances, MHC class II+ T cell clones (TCC) have been shown to present peptides and selected antigens interacting with antigen-specific TCC in the absence of APC . In this report we studied the capacity of SAg to mediate a T-T cell interaction, investigating the TCC ability to present a panel of staphylococcal enteroxins (SE) independently of the presence of added APC . Upon exposure to a broad range of SE concentrations, MHC class II+ TCC showed an intense proliferative response even in the absence of professional APC . Diverse SE optimally stimulated responder TCC at different concentrations . The proliferation was inhibited by anti-DR monoclonal antibodies, both in the presence and in the absence of APC . The SE activation of TCC in the absence of APC induced the same series of phenotypic variations as that observed following the TCC stimulation with APC . Irradiated TCC efficiently presented membrane-bound SE to responder TCC as well as professional APC . These results show that a single cell of a given clone effectively presents the SE to other cells of the same clone, and provide evidence that SAg can efficiently mediate T-T cell interaction . In addition, the possibility also exists that one cell of the clone can actually undergo an auto-stimulation via SAg-mediated interactions between its own TcR and MHC class II molecule . It has recently been suggested that the V beta-selective depletion of T cells observed in acquired immunodeficiency syndrome (AIDS) patients might be a consequence of the interaction between a human immunodeficiency virus (HIV)-encoded SAg and T cells expressing a SAg complementary V beta . We suggest that the hypothesized HIV-encoded SAg might mediate T-T cell interactions that could play a relevant role in the V beta-selective depletion of T lymphocytes observed in HIV-infected patients. J Immunol, 1992 Aug 1, 149(3), 887 - 96 T cell receptor alpha-chain influences reactivity to Mls-1 in V beta 8.1 transgenic mice; Smith HP et al.; Most, but not all, V beta 8.1+ T cells respond to M1s-1 and are clonally deleted in the thymus of M1s-1-expressing animals . To formally examine the role of the TCR alpha-chain in reactivity and tolerance to M1s-1, we have analyzed M1s-1 reactivity in a large panel of CD4+ hybridomas generated from TCR V beta 8.1 transgenic mice, that express an identical, potentially M1s-1-reactive beta-chain . The data show that the alpha-chain strongly influences the M1s-1 reactivity of the hybridomas and that the differences in reactivity had relevance for tolerance . Thus, V alpha 11+ hybridiomas were biased toward M1s-1 reactivity and V alpha 11+ T cells were correspondingly absent from the peripheral repertoire of M1s-1-expressing transgenic mice . V alpha 2+ hybridomas, on the other hand, were biased against M1s-1 reactivity, and V alpha 2+ T cells were correspondingly amplified in the M1s-1-expressing transgenic mice . Structural analysis of the alpha-chains revealed that the M1s-1 reactivity of the V alpha 11+ hybridomas segregated precisely with family member, such that V alpha 11.1+ hybridomas were M1s-1-reactive and V alpha 11.3+ hybridomas were not M1s-1-reactive . On the other hand, there was not a clear correlation between family member and M1s-1 reactivity in the V alpha 2+ hybridomas . The hybridomas also showed striking variation in their reactivity to staphylococcal enterotoxin B (SEB), and the SEB reactivity of the V alpha 11+ hybridomas correlated precisely with family member and with M1s-1 reactivity . In contrast, there was not a clear correlation with V alpha 2+ alpha-chain structure and SEB reactivity . Also, there was no correlation between M1s-1 reactivity and SEB reactivity in individual V alpha 2+ hybridomas, suggesting that the recognition of the two superantigens by the same TCR is not equivalent . Taken together, these data define a role for the TCR alpha-chain in superantigen reactivity and T cell tolerance, and provide a structural explanation for the different fates of M1s-1-reactive T cells in normal and transgenic mice. Biochemistry, 1992 Jul 21, 31(28), 6396 - 401 Comparison of conformational features of staphylococcal nuclease in ternary complexes with pdTp, pdGp, and nitrophenyl-pdTp; Stanczyk SM et al.; The conformations of wild-type staphylococcal nuclease (SNase) in the ternary complexes with thymidine 3',5'-bisphosphate (pdTp), 2'-deoxyguanine 3',5'-bisphosphate (pdGp), and thymidine 3'-phosphate 5'-(p-nitrophenylphosphate) (NpdTp) with Ca2+ were examined by two-dimensional NMR NOESY and ROESY experiments . The results of these experiments indicate that the conformational features of the SNase are quite similar in the three ternary complexes . This suggests that the conformational features of SNase, in these ternary complexes, are not strongly dependent on whether the 5'-phosphate is a mono- or diester . This is in contrast to our prior studies on substitutions of active site charged amino acids which indicated that the conformational features of SNase in the ternary complex are quite sensitive to substitutions for active site charged amino acids (Hibler et al., 1987; Wilde et al., 1988; Pourmotabbed et al., 1990) . The similarity of the SNase conformational features in the ternary complexes with pdTp and pdGp indicates that the features of the nucleotide bound at the active site are not strong determinants of the enzyme conformation in the ternary complexes . These conclusions are in general agreement with the results on pdApdT ternary complexes with SNase which suggested that it is the conformational features of the bound nucleic acid which determine the differences in catalysis observed for SNase with different substrates (Weber et al., 1991), more so than the conformational features of the enzyme. Proc Natl Acad Sci U S A, 1992 Jul 15, 89(14), 6383 - 7 Conformation-dependent cleavage of staphylococcal nuclease with a disulfide-linked iron chelate; Ermacora MR et al.; We report the synthesis and evaluation of (EDTA-2-aminoethyl) 2-pyridyl disulfide . By using this easily prepared cysteine-specific hydrophilic reagent, an ethylenediaminetriacetic acid-Fe3+ complex (EDTA-Fe) was covalently attached to a single genetically engineered cysteine residue in staphylococcal nuclease . Upon addition of the iron reductant ascorbate, the nuclease-EDTA-Fe conjugate underwent a protein self-cleavage reaction mediated by reactive oxygen species . Sequence analysis of the products indicated that cleavage occurs close in tertiary structure to the EDTA-Fe attachment site . In the presence of denaturants, the cleavage pattern changes and the reaction is limited to residues proximal in sequence to the cysteine attachment site . These results indicate that intramolecular protein cleavage reactions mediated by EDTA-Fe can be used to evaluate changes in protein conformation . The reagent described should be a useful tool in the structural mapping of nonnative protein states populated at equilibrium, such as the molten globule, that are frequently refractory to conventional structure analysis. J Biol Chem, 1992 Jul 15, 267(20), 13958 - 63 Cloning and expression of wild-type and mutant forms of the cardiotonic polypeptide anthopleurin B; Gallagher MJ et al.; Venom of the sea anemone Anthopleura xanthogrammica contains a minimum of three polypeptide toxins capable of prolonging the repolarization phase of the action potential . A synthetic gene for the most toxic of the Anthopleura toxins, anthopleurin B (ApB), has been designed, synthesized, and expressed as a fusion protein with the gene 9 product of bacteriophage T7 in Escherichia coli . The fusion protein has been purified and its disulfide bonds reoxidized using glutathione redox couples . Upon cleavage with staphylococcal protease, this protocol yields approximately 1 mg of native ApB/liter of original culture . The recombinant protein has been shown to be identical to natural ApB with respect to amino acid composition, amino-terminal sequence, secondary structure, high pressure liquid chromatographic mobility, and biological activity . A second form of ApB containing additional residues of glycine and arginine at its amino terminus has also been characterized . This protein, termed GR-ApB, is identical in specific activity to the wild-type form . This work lays the groundwork for a detailed analysis of ApB structure and action by site-directed mutagenesis. Clin Orthop, 1992 Jul, (280), 289 - 99 Osteomyelitis experimentally induced with Bacteroides thetaiotaomicron and Staphylococcus epidermidis . Influence of a foreign-body implant; Mayberry-Carson KJ et al.; Experimental osteomyelitis was induced in the rabbit tibia with Staphylococcus epidermidis alone, with Bacteroides thetaiotaomicron alone, and with both bacteria as etiologic agents, in the presence or absence of a foreign-body implant . Animals were monitored by clinical observation and roentgenographic, microbiologic, histologic, immunofluorescent microscopic, and electron microscopic methods . Scanning and transmission electron microscopy showed masses of coccoid and rod-shaped bacteria embedded in a matrix of exopolysaccharide and adhered to bone, marrow, and the foreign-body implant (when present) . Of the 58 rabbits receiving an implant, osteomyelitis developed in 48 (83%), and bacteria were recovered by culture from 56 (97%) . Of the 31 animals without the implant, osteomyelitis developed in 18 (58%), but no bacteria were recovered by culture . Bacterial recovery appeared to be dependent on the presence of the implant . The rate of induction and the severity of osteomyelitis were enhanced by the presence of the foreign-body implant and by the polymicrobic infection. Cornell Vet, 1992 Jul, 82(3), 233 - 46 Effects of transport on constituents of bronchoalveolar lavage fluid from horses; Crisman MV et al.; To determine whether road transport affected pulmonary phagocyte activity, 7 healthy Thoroughbred horses were shipped 1,160 kilometers over 36 hours . Fluid collected by bronchoalveolar lavage (BAL) 12 hours, and 7 and 14 days after transport was analyzed . Results were compared to those from the same horses pre-transport, and 7 non-transported control horses that had BAL performed at the same times as the transported horses . Of cells recovered with BAL the percentage of viable pulmonary alveolar macrophages (PAMs) declined from 90.0 +/- 0.9% pre-transport to 80.0 +/- 3.7% by 2 weeks post transport . Although the ability of PAMs to inhibit the growth of Staphylococcus epidermidis had decreased by 2 weeks post-transport (19.2 +/- 3.7% vs . 8.8 +/- 2.3% inhibition) this could not be attributed to transport as a similar effect occurred in the control group . In contrast, the ability of PAMs to phagocytose sheep erythrocytes labelled with rabbit anti-erythrocyte antibodies increased from 74.0 +/- 8.1% to 92.3 +/- 1.5% by 12 hours post-transport . As all variables were unchanged or only mildly altered following transport, we conclude that this form of transport did not alter the PAM functions we assessed. Eur J Immunol, 1992 Jul, 22(7), 1935 - 8 In vivo responses of CD4+ and CD8+ cells to bacterial superantigens; Herrmann T et al.; Staphylococcal enterotoxin B (SEB) is a bacterial superantigen that binds to major histocompatibility complex (MHC) class II molecules and specifically activates T cells bearing V beta 8 T cell receptor domains . We have compared several aspects of the response of CD4+ and CD8+ T cell subsets to SEB in vivo . V beta 8+ cells in both subsets proliferated to a similar extent upon SEB injection . Furthermore, mRNA for interferon-gamma was induced in both subsets with similar kinetics and SEB dose-response . Finally CD8+ (but not CD4+) T cells from SEB-injected mice exhibited SEB-specific lysis of MHC class II-bearing target cells . Collectively, these data indicate that the CD4: MHC class II interaction confers no detectable selective advantage to CD4+ cells in the in vivo response to SEB . The observed effector functions of both subsets may contribute to SEB-induced immunopathology. J Vasc Surg, 1992 Jul, 16(1), 75 - 86 Thoracoabdominal aortic aneurysm associated with umbilical artery catheterization: case report and review of the literature; Cribari C et al.; Aneurysms in infants and children are rare and are usually associated with cardiovascular malformations or connective tissue disorders . A new subgroup of patients has become recognized over the past two decades--those with aneurysms associated with umbilical artery catheterization . Critically ill newborns who have required umbilical artery catheterization and have developed sepsis, usually staphylococcal, are at risk for the development of mycotic aneurysm disease of the aorta or its major branches or both . Since first described in 1970, 34 cases have been reported in the literature, 14 involving the descending thoracic aorta, 10 the abdominal aorta, 6 the iliac arteries, and 4 either the thoracoabdominal aorta or multiple aneurysms involving both the thoracic and abdominal aorta . This report presents a case we recently treated of a 15-month-old-boy with a large thoracoabdominal aortic aneurysm and aneurysms of the infrarenal abdominal aorta and proximal right common iliac artery . It includes a review of the recent literature to analyze pathogenesis, clinical manifestations, and to formulate methods of treatment. J Exp Med, 1992 Jul 1, 176(1), 37 - 46 Cyclosporin A markedly enhances superantigen-induced peripheral T cell deletion and inhibits anergy induction; Vanier LE et al.; Cyclosporin A (CsA) is a well-known immunosuppressive agent that modulates immune tolerance in many ways . CsA can give rise to a state of long-term nonimmunosuppressed transplantation tolerance, but it can also aggravate autoimmune diseases, and provoke specific forms of autoimmunity . These effects, which are often paradoxical, remain largely unexplained . In this study, we investigated the effects of CsA on superantigen (superAg)-reactive peripheral T cells . The intravenous injection of either staphylococcal enterotoxin B (SEB), or Mls-1a cells into Mls-1b recipients, causes long-term in vitro nonresponsiveness (anergy) and partial elimination of the peripheral T cell receptor (TCR) V beta 8+/CD4+ and -V beta 6+/CD4+ T cell subsets, respectively . We report that CsA markedly enhances the peripheral elimination of SEB- and Mls-1a-reactive T cells such that up to 90% of the targeted CD4+/V beta subpopulations are deleted . The degree of deletion depends on the dose and the schedule of CsA administration, and the number of superAg injections . In situations where the extent of deletion is only moderate, we find that the remaining superAg-reactive T cells fail to develop anergy, unlike the T cells of control superAg-immunized mice . Higher doses of CsA are required to enhance T cell deletion (greater than or equal to 25 mg/kg/d, i.p.) than to impair anergy induction (greater than or equal to 6.25 mg/kg/d, i.p.) . In view of these results, it appears that the degree of tolerance in CsA/superAg-treated mice depends on the balance between these opposing effects, i.e., enhancement of peripheral elimination versus the abrogation of anergy . The possibility of enhancing or preventing immune tolerance with a drug may have important clinical implications. Infect Immun, 1992 Jul, 60(7), 2612 - 8 Induction of tumor necrosis factor and interleukin-1 by purified staphylococcal toxic shock syndrome toxin 1 requires the presence of both monocytes and T lymphocytes; See RH et al.; Highly purified staphylococcal toxic shock syndrome toxin 1 (TSST-1) was tested for its ability to induce the cytokines tumor necrosis factor (TNF) and interleukin-1 (IL-1) from fractionated human peripheral blood mononuclear cells prepared from seven healthy donors . Highly purified monocytes alone or T lymphocytes alone did not produce TNF or IL-1 when incubated with TSST-1 at 37 degrees C for up to 72 h . However, the addition of 10 micrograms of TSST-1 per ml to a 1:1 mixture of monocytes and T cells resulted in significant TNF (predominantly TNF-alpha) and IL-1 beta production after 24 h at 37 degrees C . The nature of the monocyte/T-cell interaction did not appear to involve gamma interferon (IFN-gamma), since 10 micrograms of rabbit anti-IFN-gamma per ml did not neutralize TNF-alpha production after TSST-1 induction . Similarly, L243, a monoclonal antibody to HLA-DR which blocks TSST-1 binding to monocytes, did not inhibit TNF-alpha production following TSST-1 induction . However, direct contact between monocytes and T cells was required, since physical separation of cells in double-chamber culture wells abolished TNF-alpha secretion after TSST-1 stimulation . Furthermore, paraformaldehyde fixation of either monocytes or T cells prior to addition to viable T cells or monocytes, respectively, also abolished TNF-alpha secretion, suggesting that aside from cell contact, soluble factors were also involved . Our results suggest that cytokine production involves more than binding of TSST-1 to its receptor on monocytes alone and that cell contact with T cells and the release of a soluble factor(s) other than IFN-gamma may be essential for the induction of cytokines by this toxin. Blood, 1992 Jul 1, 80(1), 241 - 8 Characterization of multiple quinine-dependent antibodies in a patient with episodic hemolytic uremic syndrome and immune agranulocytosis; Stroncek DF et al.; A 23-year-old woman experienced six distinct episodes of severe combined neutropenia and thrombocytopenia . At least one of the episodes was accompanied by hemodialysis-requiring acute renal failure and fragmentation hemolysis (hemolytic uremic syndrome {HUS}) . In retrospect, all episodes were probably associated with the ingestion of quinine . Quinine-dependent antibodies to platelets, neutrophils, T lymphocytes, and red blood cells (RBCs) were detected in the patient's serum . By a monoclonal antibody antigen capture assay, the patient's serum contained IgG antibodies, which in the presence, but not absence, of quinine reacted with platelet glycoprotein (GP) complexes Ib/IX and IIb/IIIa, but not Ia/IIa . By immunoprecipitation assay, the serum, after addition of quinine, reacted strongly with an 85-Kd neutrophil membrane protein and weakly with 130- and 60-Kd moieties . Serum adsorbed with RBCs in the presence of quinine continued to react with platelets and neutrophils, and serum that was absorbed with platelets continued to react with neutrophils and RBCs, indicating that the antigenic targets were different on platelets, neutrophils, and RBCs . Since platelets and endothelial cells share some antigens, we tested patient serum for antibodies to human umbilical vein endothelial cells (HUVEC); no quinine-dependent antibodies to HUVEC were detected . However, her quinine-dependent antibodies not only bound to platelets and neutrophils, but also activated neutrophils . Thus, the patient's serum with quinine aggregated neutrophils, but neither agent alone caused activation . Moreover, the patient's serum with quinine (but not without) augmented the adherence of neutrophils to HUVEC . Treatment of the patient's serum with staphylococcal protein A removed the quinine neutrophil aggregation cofactor, suggesting it was due to IgG . In both neutrophil aggregation and adherence assays, decomplementation of the patient's serum markedly blunted its effect . Furthermore, the patient's serum failed to aggregate formalin-inactivated neutrophils, suggesting neutrophil activation, probably by activated complement, was necessary for aggregation and adhesivity to endothelium . We conclude that our patient's neutropenia, thrombocytopenia, lymphopenia, and anemia were due to quinine-dependent antibodies, and that these antibodies recognized epitopes that were different in the three target cell populations . We further suggest that HUS was likely secondary to the activation and adhesion of neutrophils to endothelium. J Neurosurg, 1992 Jul, 77(1), 29 - 36 Infection of cerebrospinal fluid shunts in infants: a study of etiological factors; Pople IK et al.; The aim of this study was to find reasons for the high incidence of cerebrospinal fluid shunt infections seen in neonates . Four-hundred sixty-six consecutive shunt operations were analyzed retrospectively in 294 children, and 60 children were studied prospectively by quantitative sampling of skin bacteria before surgery and by sampling open wounds, shunt catheters, surgical gloves, and airborne bacteria . In total, 110 strains of coagulase-negative Staphylococcus isolated from the skin of 53 children before surgery were then tested for bacterial adherence . Retrospectively, the infection rate for infants younger than 6 months old was 15.7% (28 of 178 procedures), compared with 5.6% (16 of 288 procedures) for older children (p = 0.0005) . Of all infections, 67% were due to coagulase-negative Staphylococcus . Age was the only major factor influencing the infection rate . Three of the 60 children studied prospectively developed postoperative shunt infections . All were younger than 6 months and all had high skin bacterial densities before surgery . Contamination during surgery was generally low, but correlated with the preoperative skin bacterial density . Strains of coagulase-negative Staphylococcus with high bacterial adherence were more commonly found in neonates than in older children . High skin bacterial density in neonates before surgery was a risk factor for infection in this study . These results also suggest that there is selection of more virulent strains of coagulase-negative Staphylococcus on the skin of neonates . Prevention of shunt infections in this high-risk group could be facilitated by the reduction of skin bacterial density before surgery using chlorhexidine shampoos and by the elimination of contamination by skin bacteria during surgery using packs soaked in an antiseptic agent to isolate wound edges and glove-changing before handling the shunt. J Immunol, 1992 Jul 1, 149(1), 317 - 22 Staphylococcal enterotoxin-mediated human T-T cell interactions; Koning F et al.; Staphylococcal enterotoxins (SE) are known to stimulate a large proportion of T cells . SE bind to MHC-class II molecules on APC and a particular segment of certain TCR V beta and V gamma gene products . Resting human T cells do not express HLA class II Ag and therefore cannot present SE to T cells . Activated human T cells, however, do express HLA-DR, -DP, and -DQ Ag and could consequently serve as APC for SE . As such, local immune responses to SE might be regulated and/or abrogated by SE-mediated T-T cell interactions leading to T cell destruction . We have investigated if such SE-mediated T-T cell interactions can occur in vitro using human cytolytic TCR-alpha beta+ and TCR-gamma delta+ T cell clones . We demonstrate that the TCR-alpha beta+ T cell clones can efficiently present staphylococcal enterotoxin A (SEA) to each other: T cell clones coated with SEA are lysed by SEA-reactive T cell clones but not by a SEA-nonreactive T cell clone . Furthermore, the SEA-reactive TCR-alpha beta+ clones (but not the SEA-nonreactive clone) destruct themselves in the presence of SEA at low concentrations of SEA (less than 0.01 microgram/ml) . Also, SEA-coated T cell clones can induce proliferative responses although such responses are much weaker than those induced when B cells are used as stimulator cells . In contrast, the SEA-reactive TCR-gamma delta+ T cell clones are resistant to autokilling in the presence of SEA and they do not lyse SEA-coated TCR-gamma delta+ targets . However, such targets can be lysed by TCR-alpha beta+ effector cells . These results indicate that TCR-gamma delta+ cells are relatively resistant to lysis and that during local nonspecific immune responses triggered by SE, which induces HLA-class II expression by the responding T cells, SE-mediated T-T cell interactions may play a role in the regulation and/or abrogation of these immune responses. Dtsch Tierarztl Wochenschr, 1992 Jul, 99(7), 306 - 10 {The effect of bacterial interference phenomena in experimental Staphylococcus hyicus infections of gnotobiotic piglets}; Waldmann KH et al.; The prevention of exudative epidermitis could be confirmed in experimental investigations with gnotobiotic piglets when the skin first was colonized with avirulent strains of Staphylococcus (Staph.) hyicus and subsequently exposed to virulent strains of Staph . hyicus . However, locally restricted cutaneous lesions in the area of application corresponding to exudative epidermitis were seen in five of nine piglets . Using the strain Staph . sciuri the spread of virulent Staph . hyicus could not be suppressed . Such infected two piglets developed generalized exudative epidermitis . In another experiment with four piglets it could be shown, that the relative protective mechanism correlating to bacterial interference on the one hand can be influenced by the virulence of causative organisms . On the other hand it even can be abolished when skin lesions are involved . For that reason probably the utilization of bacterial interference in prevention of exudative epidermitis under field conditions is considerably limited. J Vet Diagn Invest, 1992 Jul, 4(3), 270 - 8 Pleuropneumonia caused by Actinobacillus pleuropneumoniae biotype 2 in growing and finishing pigs; Frank RK et al.; Actinobacillus pleuropneumoniae biotype 2 was isolated in pure culture or as the predominant isolate from the lungs of 9 growing and finishing pigs with pleuropneumonia . Gross and microscopic lesions resembled those caused by A . pleuropneumoniae biotype 1 serotypes (Nos . 1, 5, and 7) traditionally seen in the United States . The overall mortality rate for growing and finishing pigs on this 1,200-sow farrow-to-finish farm ranged from 0.37% to 0.84% per month from July 1990 to February 1991, and mortality due to respiratory disease ranged from 0.17% to 0.52% per month for the same period . This Actinobacillus species did not require V factor (no satellitism on blood agar with a Staphylococcus streak), was strongly beta-hemolytic, and demonstrated restriction fragment length polymorphisms in hybridization studies with A . suis, A . lignieresii, and A . equuli . Biochemically, the isolate most closely resembled A . pleuropneumoniae, and a DNA fragment considered specific for A . pleuropneumoniae biotypes 1 and 2 was demonstrated using polymerase chain reaction . Necrohemorrhagic pleuropneumonia similar to that caused by A . pleuropneumoniae biotype 1 was reproduced experimentally in 2 4-week-old pigs inoculated intratracheally with broth cultures of the A . pleuropneumoniae biotype 2 . This study demonstrated the presence of A . pleuropneumoniae biotype 2 in the United States. FEMS Microbiol Immunol, 1992 Jul, 4(5), 247 - 54 Murine monoclonal antibodies against staphylococcal enterotoxin B: production and characterization; Goyache J et al.; A group of 14 monoclonal antibodies (mAbs) to staphylococcal enterotoxin B (SEB) were obtained by fusion of Sp2/O myeloma cells with spleen cells from female BALB/c mice immunized with commercial SEB . The antibodies belonged to IgG1 and IgG2b subclasses . We evaluated the anti-SEB titres, competition assays and sensitivity of detection by indirect ELISA . Reactivity and cross-reactivity were also studied by indirect ELISA and confirmed by immunoblotting . All the mAbs reacted with SEB and with a second band which had a different electrophoretic mobility and probably represents an aggregate of SEB or SEB bound to membranes . Three mAbs reacted only with SEB and the rest showed cross-reactions with SEC1 . No reactions were observed against any other serovar (SEA, SED and SEE) or other proteins. Anal Biochem, 1992 Jul, 204(1), 26 - 33 A method for the evaluation of the efficiency of signal sequences for secretion and correct N-terminal processing of human parathyroid hormone produced in Escherichia coli; Kareem BN et al.; Expression plasmids have been constructed for evaluation of different signal sequences for secretion and correct amino terminal processing of foreign proteins expressed in