Microbiol Immunol, 1996, 40(2), 115 - 9 Detection and measurement of Staphylococcus epidermidis slime using an ELISA technique; Mempel M et al.; A polyclonal rabbit anti-serum against the strong slime-producing Staphylococcus epidermidis strain RP62A was absorbed with the slime-negative phase variant of this strain PV1 in order to remove not slime-specific antibodies . Using this antiserum we established an ELISA which enables detection of slime production in S . epidermidis extracts . The ELISA showed high absorbance when extracts from slime-positive strains (confirmed in the tissue culture tube test) were used as antigens . The high absorbance of slime-positive strains was greatly reduced by periodate oxidation of the extracts and was resistant to proteinase digestion suggesting that the detected antigen is composed of polysaccharides . In contrast to other rapid and simple laboratory detection methods for S . epidermidis slime, the slime-specific ELISA gave positive results in the presence of human serum. Beitr Infusionsther Transfusionsmed, 1996, 33, 191 - 5 Influence of hemodilution on cefuroxime levels and bacterial contamination of intra- and postoperative processed wound blood during hip replacement; Wollinsky KH et al.; MATERIALS AND METHODS: In a prospective randomized study we investigated the effect of hemodilution on cefuroxime levels and bacterial contamination of processed shed blood during hip arthroplasty . 10 patients received cefuroxime 1,5 g as single shot prophylaxis before (group A), 10 after hemodilution (15 ml/KG) (group B) . Cefuroxime levels in serum 1 h after administration, at the end of operation, after 12 h and in drainage-blood after 12 h were assessed by HPLC . Bacteriological study was performed from collecting bags (Vacufix), wound drainage blood and processed red blood cell concentrates, using pour plate technique and broth culture enrichment . RESULTS AND DISCUSSION: Mean concentrations of cefuroxime were higher in group B than group A, but differed not significantly . No bacterial contamination was found in collecting bags and wound drainages in both groups . Processed red cell concentrates in group B showed no growth . In group A, however, 3/10 were contaminated with < or = CFU/ml of coagulase-negative Staphylococcus, Staphylococcus epidermidis and Propionibacterium acnes . The differences between groups did not reach the level of statistical significance and could not be related to lower cefuroxime levels . No wound infection occurred in either group. ASAIO J, 1996 Jan-Feb, 42(1), 52 - 9 Synthesis of tumor necrosis factor alpha and interleukin-1 receptor antagonist, but not interleukin-1, by human mononuclear cells is enhanced by exposure of whole blood to shear stress; Pomianek MJ et al.; Extracorporeal circulation exposes blood to shear stress . In many studies, researchers reported effects of shear stress on morphology and function of various blood cells, but effects on cytokine synthesis have not been studied . The authors investigated the effect of shear stress on the synthesis of interleukin-1 beta, interleukin-1 alpha, tumor necrosis factor alpha, and interleukin-1 receptor antagonist by human peripheral blood mononuclear cells . Whole heparinized blood at room temperature was exposed to shear stresses of 50, 200, or 500 dyne/cm2 for 5 min or 30 sec, and to 980 dyne/cm2 for 5 sec . Peripheral blood mononuclear cells were then separated from sheared blood and cultured for 24 hrs with or without lipopolysaccharide or Staphylococcus epidermidis . Total (intra + extracellular) cytokine synthesis was measured by specific radioimmunoassay . Viability of cultured peripheral blood mononuclear cells, determined by trypan blue exclusion and lactate dehydrogenase release, was not significantly affected by shear stress . Shear stress without lipopolysaccharide or S . epidermidis stimulation did not affect synthesis of interleukin-1 or tumor necrosis factor alpha but did enhance synthesis of interleukin-1 receptor antagonist . Lipopolysaccharide- or S . epidermidis- induced synthesis of interleukin-1 was not significantly altered by shear stress . In contrast, lipopolysaccharide-induced tumor necrosis factor alpha synthesis increased with increasing shear stress and was significantly elevated over unsheared controls, whereas S . epidermidis-induced tumor necrosis factor alpha and lipopolysaccharide- or S . epidermidis-induced interleukin-1 receptor antagonist synthesis were not significantly enhanced by shear . Therefore, sublytic trauma, such as exposure to shear stress, affects in vitro responses of peripheral blood mononuclear cells to secondary stimuli. Retina, 1996, 16(3), 246 - 9 Intravenous cefazolin in penetrating eye injuries . A swine model; Nossov PC et al.; PURPOSE: A swine model of penetrating ocular trauma was used to determine the delivery of systemically administered cefazolin to the vitreous cavity of traumatized and nontraumatized eyes . METHODS: Thirty-one pigs received a scleral laceration to the right eye under anesthesia and then were given intravenous cefazolin every 8 hours . Seven pigs received nine doses at 17 mg/kg . Seven animals received three doses of 36 mg/kg, and six others received nine doses of this regimen . Six pigs received three doses of 79 mg/kg and five others received three doses of 190 mg/kg . RESULTS: Vitreous levels of cefazolin averaged 15.6 micrograms/mL in traumatized eyes but were less than 1 microgram/mL in control eyes of animals receiving three doses at 190 mg/kg (P < or = 0.025) . Mean serum concentration in these animals was 49.3 micrograms/mL . Vitreous levels were less than 1 microgram/mL in traumatized and control eyes in animals given lower doses of cefazolin (range, 17-79 mg/kg) despite multiple treatments over 2 and 3 days . CONCLUSIONS: These data demonstrate that systemically delivered cefazolin achieves levels ten times the minimum inhibitory concentration for Staphylococcus epidermidis in injured eyes . Therapeutic intraocular levels can be obtained through intravenous dosing, provided that therapeutic serum concentrations are achieved. Antimicrob Agents Chemother, 1996 Jan, 40(1), 215 - 7 Tandem duplication in ermC translational attenuator of the macrolide-lincosamide-streptogramin B resistance plasmid pSES6 from Staphylococcus equorum; Lodder G et al.; A tandem duplication of 23 bp in the ermC gene translational attenuator of plasmid pSES6 from Staphylococcus equorum which mediated constitutive resistance to macrolide-lincosamide-streptogramin B antibiotics was identified . This duplication included the ribosome binding site for the ermC gene as well as the first 5 bp of the ermC coding sequence . It was postulated that this sequence duplication affects the possible RNA conformations so that the ribosome binding site for ErmC synthesis is readily accessible to the ribosomes and thus constitutive expression of the ermC gene occurs. Br J Dermatol, 1996 Jan, 134(1), 120 - 2 The occurrence of bacteraemia with skin surgery; Carmichael AJ et al.; The need for chemoprophylaxis for bacterial endocarditis is partly dependent on the risk of bacteraemia associated with the procedure, which has not been adequately defined for skin surgery . The incidence of postoperative bacteraemia in 149 immunocompetent out-patients with non-infected lesions was 0.7% (95% CI 0.3-3.8%) . Procedures included excisions, flaps, grafts and micrographically controlled surgery . Coagulase-negative staphylococcus was the most common skin isolate at the site of surgery, present in 68.5% of patients . The most effective chemoprophylaxis would be intravenous vancomycin, which is inconvenient and has an inherent risk of morbidity . Given the low incidence of bacteraemia and the disadvantages of the optimum chemoprophylaxis, surgery on non-infected lesions does not warrant prophylactic antibiotics to prevent the very low risk of bacterial endocarditis. Vet Res Commun, 1996, 20(3), 205 - 14 The antimicrobial susceptibility of Staphylococcus species isolated from canine dermatitis; Kruse H et al.; In a retrospective study, 1538 strains of beta-haemolysin-producing Staphylococcus species isolated from dermatitis in dogs at three veterinary clinical microbiology laboratories in Norway during 1986-87 and 1993-94 were investigated for their antimicrobial susceptibility . None of the strains was resistant to cloxacillin, cephalexin or the quinolones enrofloxacin and ciprofloxacin . More than 96% of the strains were susceptible to trimethoprim-sulphonamide, bacitracin and fucidic acid . Between 67% and 89% of the strains were susceptible to erythromycin, lincosamides, tetracycline, neomycin and chloramphenicol . Only 37.9% of the strains were susceptible to penicillin . The frequency of penicillin resistance increased significantly between the first and second periods, from 46.0% to 58.6% . The frequency of resistance to lincomycin, clindamycin and erythromycin also increased significantly between the first and second periods, from 3.0%, 2.1% and 3.3% to 25.5%, 19.5% and 24.8%, respectively . A moderate increase in resistance to tetracycline was also noted, from 20.4% in the first to 27.6% in the second period . On the other hand, the frequency of resistance to trimethoprim-sulphonamide decreased significantly from 4.1% in the first to 0.9% in the second period . Many different resistance patterns were observed in each period . However, the proportion of multiresistant strains increased from 2.1% in the first to 10.2% in the second period . There was a decrease in resistance to the combination of trimethoprim-sulphonamide and penicillin from the first to the second period . Resistance to the combination of lincosamides and penicillin increased . For the combinations penicillin-tetracycline-lincosamides, penicillin-lincosamides-erythromycin, and penicillin-tetracycline-lincosamides-erythromycin, there was a striking increase in resistance between the first and the second periods. Ann Fr Anesth Reanim, 1996, 15(2), 189 - 91 {Prolonged apnea after suxamethonium administration during staphylococcal toxic shock}; Blanloeil Y et al.; A toxic shock syndrome occurred after a femoral nail removal requiring revision surgery . After administration of suxamethonium (1 mg.kg-1), an apnoea prolonged over 45 minutes was observed . The trachea was extubated 105 minutes after suxamethonium administration . For the nail removal, two days before, the anaesthetic had been given by the same anaesthesiologist, with a similar protocol . Apnoea extended over 20 minutes . The day of the revision surgery, plasma cholinesterase activity was 410 UI.L-1 and reached 910 UI.L-1, 9 months later . Dibucaine number was 20 and fluorure number 17 . The apnoea was in relation with a genetic plasma cholinesterase deficiency increased by the toxic shock syndrome . Shock and hepatic insufficiency were suspected to contribute to the decrease in plasma cholinesterase . Suxamethonium should be avoided in case of toxic shock syndrome. Int J Immunopharmacol, 1996 Jan, 18(1), 79 - 87 Comparative evaluation of in vitro and in vivo immunosuppressive potential of cyclosporin G with cyclosporin A and FK-506; Saada V et al.; Cyclosporin G (CsG), a promising cyclosporin A (CsA) analogue, was examined and compared with two reference immunosuppressive drugs: CsA and FK-506, regarding their inhibitory effects on different lymphocyte activation pathways as well as on graft-versus-host reaction (GvHR) across differences at major or minor histocompatibility loci . The results showed that, at different concentrations, CsG efficiently inhibited proliferation induced by alloantigens (mixed lymphocyte culture), mitogens (concanavalin A, pokeweed mitogen) and the combination of phorbol myristate acetate + ionomycin, to the same extent as observed with CsA and FK-506 . It was also shown that CsG exhibited the same strong inhibitory effects as the two other immunosuppressants upon stimulation triggered by viral (MLs-1a) or bacterial (staphylococcal enterotoxin B) superantigen . Determination of IL-2 activity in the supernatant of MLC also confirmed similar strong inhibitory effects, exerted by CsG compared to CsA and FK-506 . In systemic and local GvHR across major or minor histocompatibility barriers, CsG as well as CsA and FK-506 presented an equivalent immunosuppressive potential . In conclusion, from various experiments involving different modes of activation, it was shown that CsG was as strongly immunosuppressive as CsA and FK-506. Free Radic Biol Med, 1996, 20(6), 813 - 20 Activation of the HIV long terminal repeat and viral production by H2O2-vanadate; Kazazi F et al.; The long terminal repeat (LTR) of human immunodeficiency virus type 1 (HIV-1) contains sequences required for the initiation of gene transcription . Among the substances known to activate the HIV-1 LTR is hydrogen peroxide (H2O2) . We report here that H2O2-induced activation of the LTR in the macrophage cell line THP-1 and the lymphocyte cell line, Jurkat, is greatly increased by vanadate . Activation of the LTR by phorbol myristate acetate, tumor necrosis factor alpha, lipopolysaccharide, or Staphylococcus epidermidis extract was not increased by vanadate, indicating some selectivity for H2O2 . H2O2 and vanadate also acted synergistically to increase the production of HIV-1 virions by the latently infected macrophage cell line U-1 as determined by p24 antigen release and the detection of intact virions by electron microscopy . Effects were observed at H2O2 and vanadate concentrations down to 3 x 10(-6) M, with high concentrations leading to cell toxicity . Catalase was strongly inhibitory when added prior to the interaction of H2O2 and vanadate, but was considerably less inhibitory when the H2O2 and vanadate were allowed to preincubate prior to the catalase addition . H2O2 reacts with vanadate to form peroxides of vanadate that have potent biological effects . Our findings suggest that among these is the activation of the HIV-1 LTR. Biol Neonate, 1996, 69(4), 249 - 56 Staphylococcus epidermidis sepsis in the intensive care nursery: a characterization of risk associations in infants < 1,000 g; Johnson-Robbins LA et al.; We undertook to determine Staphylococcus epidermidis colonization patterns and risks of sepsis in a cohort of 82 consecutive intensive care nursery admissions (birth weight 1,285 +/- 57 g), with 24 infants weighing < 1,000 g at birth . Colonization was determined by skin and stool cultures collected at three time points . Multiple neonatal variables were classified into three intervals preceding the time of sample collection including the occurrence of S . epidermidis sepsis . 16 infants (20%) developed S . epidermidis sepsis . 81% of these episodes occurred in infants < 1,000 g . Skin colonization was nearly universal at all sampling points . Rectal colonization was 63.6% initially (10 +/- 0.4 days), then declined to 32% by the third sample (37 +/- 0.4 days) . Neither prevalence of skin nor rectal colonization influenced the incidence of sepsis significantly . Statistically significant risk associations for sepsis for the entire intensive care nursery population included: low birth weight, gestational age, presence of a central line, and delayed feeding . For infants < 1,000 g the occurrence of sepsis during the second study time period (54% of the episodes) was associated with preceding steroid exposure . During the third study time period, birth weight and delayed attainment of full enteral feeds showed a statistically significant association with sepsis . We conclude that infants < 1,000 g are at an increased risk of S . epidermidis sepsis . Extreme immaturity, steroid therapy, and prolonged hyperalimentation are all significant risk associations. Acta Derm Venereol, 1996 Jan, 76(1), 10 - 2 Soluble Pityrosporum-derived chemoattractant for polymorphonuclear leukocytes of psoriatic patients; Bunse T et al.; The chemoattraction of polymorphonuclear leukocytes (PMNs) from psoriatic patients, atopic patients and healthy control persons by Pityrosporum orbicularelovale was investigated using the Boyden chamber method . The chemotactical attraction of PMNs from psoriatic patients by Pityrosporum (stimulation index SI = 58 +/- 50) was significantly increased (p < 0.05) compared to PMNs from atopic patients (SI = 20 +/- 17) and control persons (SI = 26 +/- 24) . This effect seems to be specific for Pityrosporum, since the chemotactical response to Staphylococcus epidermidis was not increased in psoriasis . The chemotactical factor produced by Pityrosporum is hydrophilic and is destroyed by acid hydrolysis, indicating its protein nature . The yeast Pityrosporum may thus play a role in the koebnerization of psoriasis. J Biomol NMR, 1996 Jan, 7(1), 77 - 82 Separation of intramolecular NOE and exchange peaks in water exchange spectroscopy using spin-echo filters; Mori S et al.; A technique for separating intramolecular NOE and solvent-proton exchange peaks in exchange spectroscopy is demonstrated . This method utilizes the large differences in relaxation and coupling properties of water and macromolecules to separate the two effects . The spin-echo filter consists of a water-frequency selective 90 degrees pulse followed by a spin-echo sequence . If the echo time is sufficiently long, protein resonances (e.g . C alpha H protons) excited by the selective pulse are removed due to their much shorter T2 values and J-coupling evolution . By combining the filter with exchange spectroscopy (EXSY) or water exchange (WEX) filter experiments, exchange peaks can be selectively observed . In this paper the filter is combined with a modified version of the WEX filter (WEX II filter) with 1D and 2D detection and applied to a zinc finger peptide and to staphylococcal nuclease, allowing estimation of the contribution of intramolecular NOEs to the exchange spectra. Diabetologia, 1996 Jan, 39(1), 28 - 36 Resistance to tolerance induction in the diabetes-prone biobreeding rat as one manifestation of abnormal responses to superantigens; Sellins KS et al.; T cells taken from normal rats treated with an exogenous source of bacterial superantigen in vivo specifically failed to proliferate following re-stimulation with the same superantigen in vitro . Responsiveness was restored following the addition of an exogenous source of interleukin-2 indicating that the T cells had been made functionally tolerant and not deleted . While staphylococcal enterotoxin treatment of normal rats virtually abolished T-cell proliferation to the same enterotoxin in vitro, T cells from similarly treated diabetes-prone Biobreeding (BB-DP) rats were markedly resistant to this in vivo effect . Responses in BB-DP rats were never reduced by more than 50% even when a 4 times more effective dose of enterotoxin was employed . The resistance of BB-DP peripheral T cells to staphylococcal enterotoxin-induced tolerance could not be attributed to differences in T-cell receptor V beta chain family usage of BB-DP vs normal T cells but was associated with qualitative differences in the way in which BB-DP T cells responded to staphylococcal enterotoxins in vitro . While under optimal stimulatory conditions BB-DP T-cell proliferative responses to staphylococcal enterotoxins appeared comparable to those from non-diabetes-prone animals, under superoptimal conditions BB-DP, but not diabetes-resistant, donor T-cell proliferative responses to staphylococcal enterotoxins could be blocked in vitro with antibodies to CD4 antigens . In addition, BB-DP T-cell proliferative responses were more sensitive to suboptimal staphylococcal enterotoxin doses in vitro . We discuss ways in which abnormal BB-DP T-cell responses to superantigens in general and resistance to staphylococcal enterotoxin-mediated tolerance induction in particular may play a role in the generation of a peripheral T-cell repertoire prone to autoimmunity. Vet Microbiol, 1996 Jan, 48(1-2), 9 - 17 Correlation between occurrence of exudative epidermitis and exfoliative toxin-producing ability of Staphylococcus hyicus; Tanabe T et al.; Staphylococcus hyicus was isolated from healthy pigs and pigs affected with exudative epidermitis (EE) . Thirty seven strains (P-7 to P-43) were isolated from pigs affected with EE on 8 farms while 131 strains were isolated from healthy pigs bred on 2 farms in Japan . Isolation rate for pigs affected with EE was 100% while that for healthy pigs was 35.4% . The biochemical and cultural characteristics of the isolates from healthy and diseased pigs were identical except for the Voges-Proskauer reaction . The culture supernatant of many isolates caused skin exfoliation in 1-day-old chickens . Therefore, many isolates were considered to produce S . hyicus exfoliative toxin (shET) . The rate of shET production by the isolates from piglets affected with EE was 87.5%, while that of the isolates from healthy pigs was 76.1% . shETs were divided in two serotypes by immunodiffusion . Piglets experimentally infected with shET-producing and nonproducing strains were observed . Local skin erythema at the inoculation site was observed with nonproducing strains and disappeared within 48 h, while the skin erythema at the sites inoculated with shET-producing strains did not disappear until 7 days after inoculation . Typical clinical signs, such as exfoliation, exudation and crusting were observed only in the piglets inoculated with shET-producing strains. J Cancer Res Clin Oncol, 1996, 122(8), 458 - 64 Analysis of T cell receptor V beta expression in rabbit T lymphocytes induced to proliferate by an HTLV-I-transformed leukemogenic T cell line; Isono T et al.; Peripheral blood lymphocytes (PBL) from rabbits of the Chbb:HM strain proliferated in coculture with an X-ray-irradiated HTLV-I-transformed leukemogenic cell line of (B/J x Chbb:HM) F1 origin, whereas PBL from rabbits of the B/J and F1 strains hardly proliferated at all in co-culture with the same cell line . A proviral HTLV-I genome was detected in high-molecular-mass DNA from these proliferating cells . An analysis of T cell receptor V beta expression revealed that these lymphocytes were of restricted V beta subfamilies, suggesting that the preferential stimulation and transformation of lymphocytes occurred in this co-culture . Staphylococcal enterotoxins similarly stimulated lymphocytes and the proliferated lymphocytes were mostly of distinct V beta subfamilies depending on stimulator enterotoxins . These results suggested that the leukemogenic cell line possesses an antigen that preferentially stimulates lymphocytes of restricted V beta subfamilies. J Biomater Appl, 1996 Jan, 10(3), 210 - 6 Development of silastic polyurethane (Angioflex) materials with antibacterial agent; Rathinam K et al.; Bioimplants incorporated with antimicrobial agents are needed to control Foreign Body Associated Infection (FBAI) in clinical settings . Attempts are made here to develop five different types of polyurethane (Angioflex), viz., (1) bare polymer, (2) bare polymer glow discharged, (3) bare polymer coated with chlorhexidine, (4) chlorhexidine coated polymer glow discharged, and (5) material (4) recoated with another layer of chlorhexidine digluconate . These materials are tested for their in vitro antibacterial effects using disc diffusion technique against five different standard clinical staphylococcus strains, viz., Wood 46 (Staph . aureus), A 182 (Staph . epidermidis), A 313 (Staph . epidermidis), A 61 (Staph . epidermidis), and A 72 (Staph . epidermidis) . Maximum antibacterial effects (zone of inhibition) are observed with polyurethanes incorporated with chlorhexidine digluconate (3) and chlorhexidine incorporated and glow discharged (4) . Findings of this study indicate that glow discharge does not seem to produce either additive 8r synergistic antibacterial effects with chlorhexidine digluconate coated Angioflex material. Immunology, 1996 Jan, 87(1), 49 - 54 The superantigen Staphylococcus enterotoxin B induces a strong and accelerated secondary T-cell response rather than anergy; Schultz H et al.; The primary and secondary immune response of V beta 8+ T cells to the bacterial superantigen Staphylococcus enterotoxin B was compared in BALB/c mice . Secondary responder T cells were found to up-regulate the expression of the adhesion molecule LFA-1 faster, and to enter the cell cycle earlier than primary responder T cells . Both, primary and secondary responder T cells upregulate the expression of CD2 and CD25 and turn into blast cells with superimposable time kinetics . Secondary responder T cells terminate DNA synthesis, blast formation and the upregulation of CD25 and CD2 expression earlier than primary responder T cells and become more rapidly deleted . Two days after superantigen challenge, when primary responder T cells reach peak activity in terms of DNA synthesis and blast formation, secondary responder T cells have returned to the size of microblasts and ceased to replicate their DNA . Whereas our results are consistent with the observations leading to the concept of superantigen-induced T-cell anergy, they demonstrate, by revealing the accelerated vigorous secondary T-cell response to the superantigen, that this concept requires reconsideration. J Surg Res, 1996 Jan, 60(1), 3 - 6 Antibiotic efficacy against Staphylococcus epidermidis adherent to vascular grafts; Bergamini TM et al.; Antibiotic bonded grafts may improve the treatment results of vascular graft infections . The purpose of this study was to determine the antibiotic and antibiotic concentration needed to be effective against Staphylococcus epidermidis-infected vascular grafts . The efficacy of four antibiotics (minocycline, cefazolin, vancomycin, and rifampin) against S . epidermidis adherent to Dacron or Teflon vascular grafts was studied in vitro . Kill kinetic studies were performed with 18 and 42 hr of exposure of Dacron-adherent and Teflon-adherent S . epidermidis at 1, 4, 16, and 64 times the minimum inhibitory concentration (MIC) of each antibiotic . Antibiotic efficacy against graft-adherent S . epidermidis at 42 hr was best at concentrations 64x MIC for minocycline, cefazolin, and vancomycin and 4x MIC for rifampin . None of the antibiotics totally eradicated the graft-adherent bacteria . Antibiotics were equally effective for S . epidermidis adherent to Dacron and Teflon grafts . Antibiotic concentrations several times that predicted by the MIC were needed for all antibiotics to achieve significant killing of graft-adherent bacteria, with rifampin the most effective at the lowest concentration. J Lab Clin Med, 1996 Jan, 127(1), 71 - 80 Influence of strain, biomaterial, proteins, and oncostatic chemotherapy on Staphylococcus epidermidis adhesion to intravascular catheters in vitro; Galliani S et al.; Initial adhesion of four phenotypically different strains of Staphylococcus epidermidis to 16 silicone, polyurethane, or hydrophilic polyurethane catheters was assessed in vitro by a bacterial radiolabeling method . The effect of catheter exposure to plasma proteins, to an anticancer polychemotherapy (5-fluorouracil, doxorubicin, cyclophosphamide), or to both of them was determined . Bacterial adhesion on native catheters was dependent on the hydrophobicity of both bacteria and catheters . The four strains tested adhered preferentially to silicone catheters (p < 0.05); adhesion was moderate to polyurethane surfaces, whereas the least adhesion was obtained for hydrophilic polyurethane catheters . Adsorption of plasma proteins on the surface produced a marked decrease in adhesion on silicone (-66.2%; p < 0.001) and polyurethane (-32.8%; p < 0.01) catheters and a marked increase in adhesion on hydrophilic surfaces (+91.7%; p < 0.05) . Chemotherapeutic treatment of the catheter produced a slight but not significant decrease in adhesion on silicone (-17.4%) and polyurethane (-19.8%) catheters and a marked increase in adhesion on hydrophilic polyurethanes (+148.2%; p < 0.001) . The in vitro simulation of catheter use suggested that oncostatic drugs and plasma proteins play an important role in S . epidermidis adhesion to intravascular catheters . Overall, bacterial adhesion is lowest on hydrophilic polyurethane catheters before and after simulation of catheter use. FASEB J, 1996 Jan, 10(1), 67 - 74 Thermodynamics of denaturation of staphylococcal nuclease mutants: an intermediate state in protein folding; Carra JH et al.; A valuable approach to understanding the forces that maintain protein structure is to analyze the thermodynamic effects of mutations on protein folding . The folding process is most often described using an energetic model that assumes a two-state transition between the native and denatured states . However, some results obtained using this approach for mutants of the protein staphylococcal nuclease have contradicted expectations from our current understanding of protein energetics . The application of differential scanning calorimetry to a set of mutant nuclease proteins allowed us to measure directly the effects of mutations on the enthalpy and heat capacity changes of unfolding, as well as on the cooperativity . We found that most of these effects can be understood with a three-state model of folding including a distinct intermediate, but not with the two-state model . Use of a three-state instead of a two-state model leads to large differences in conclusions about the stability effects of some mutations, suggesting that reevaluation of the effects of mutations on this and other proteins may be necessary to achieve an accurate description of folding energetics . The two-state assumption commonly used in protein stability studies may be an oversimplification in many cases. Eur J Immunol, 1996 Jan, 26(1), 231 - 7 Activation via the antigen receptor is impaired in T cells, but not in B cells from patients with common variable immunodeficiency; Fischer MB et al.; The patients included in this study belong to a subset of common variable immunodeficiency (CVID) patients whose peripheral blood T cells have a T cell receptor (TCR)-mediated activation defect leading to impaired expression of the interleukin (IL)-2 gene upon stimulation with recall antigens (tetanus toxoid, Escherichia coli) or superantigens (staphylococcal enterotoxins) . In the present report we demonstrate that the patients' peripheral blood T cells failed to generate the second messenger inositol 1,4,5-trisphosphate (Ins(1,4,5)P3) following stimulation with superantigen or mAb specific for the monomorphic region of the TCR beta-chain . Patients' T cell lines were also impaired in generating Ins(1,4,5)P3 when stimulated with tetanus toxoid-pulsed autologous monocytes . Addition of a second or third co-stimulatory signal provided by recombinant IL-2, CD28 or both had no effect on the Ins(1,4,5)P3 formation of the patients' antigen-driven T cell lines . The T cell activation defect, however, was not absolute, as Ins(1,4,5)P3 formation in the patients' T cells after phytohemagglutinin or aluminium fluoride stimulation was normal . The impairment in signal transduction via the T cell antigen receptor was limited to the patients' T cells, as no activation defect after ligation of surface immunoglobulin, the antigen receptor on B cells, could be detected. Eur J Immunol, 1996 Jan, 26(1), 1 - 9 Antibody-targeted superantigen therapy induces tumor-infiltrating lymphocytes, excessive cytokine production, and apoptosis in human colon carcinoma; Litton MJ et al.; Bacterial superantigens are the most potent known activators of human T lymphocytes . To engineer superantigens for immunotherapy of human colon carcinoma, the superantigen, staphylococcal enterotoxin A (SEA) was genetically fused to the Fab region of the colon carcinoma-reactive monoclonal antibody C242 . In the present study the effector mechanisms involved in the anti-tumor response to C242 Fab-SEA were characterized . Immunohistochemistry and computer-aided image analysis were used in studies of cryopreserved tumor tissue to evaluate the phenotype of infiltrating cells and their cytokine profiles in response to therapy . Human T cells and monocytes were recruited to the tumor area and penetrated the entire tumor mass within hours after injection of C242 Fab-SEA . The production of cytokines at the single-cell level was found to be dominated by tumor necrosis factor (TNF)-alpha, interleukin (IL)-2, IL-4, IL-5, IL-10, IL-12, interferon (IFN)-gamma, granulocyte-macrophage colony-stimulating factor, and transforming growth factor-beta, whereas IL-1-alpha, IL-1ra, IL-1 beta, TNF-beta, IL-3, IL-6, and IL-8 were undetectable . Most of the TNF-alpha, IL-2, IL-12, and IFN-gamma were made by the infiltrating human leukocytes, while the colon carcinoma cells were induced to produce IL-4, IL-10, and TNF-alpha . Up-regulation of IFN-gamma receptors and TNF R p60 receptors was found, while the TNF R p80 receptor was absent . The cytokine production, T cell infiltration, and CD95 Fas receptor expression concomitantly occurred to induce programmed cell death in the tumor cells . This was followed by a strong reduction of the tumor mass that was seen within 24 h after C242 Fab-SEA infusion . These findings demonstrate that antibody-superantigen proteins efficiently recruit tumor-infiltrating lymphocytes actively producing a variety of cytokines likely to be essential for the therapeutic effects observed in the model . Although the humanized SCID model has obvious limitations in its predictive value for treatment of human cancer, we believe that these results encourage clinical evaluation of antibody-targeted superantigens. Ann Thorac Surg, 1996 Jan, 61(1), 359 - 65; discussion 372-3 Implantable LVAD infections: implications for permanent use of the device; McCarthy PM et al.; BACKGROUND . Infection in implantable left ventricular assist device (LVAD) patients is common and has serious implications regarding permanent use of the LVAD . METHODS . Thirty-three patients had HeartMate LVAD insertion as a bridge to heart transplantation . The mean length of hospital stay was 8 days before LVAD insertion . Before insertion 6 patients (18%) had positive pulmonary cultures and 5 patients (15%) had bacteremia . RESULTS . During LVAD support 18 patients (55%) had bloodstream infection . Of 24 patients (73%) successfully bridged to transplantation, 12 (50%) had positive blood cultures including Staphylococcus species (n = 9), Candida (n = 3), Pseudomonas (n = 2), and Enterococcus (n = 2) . Infectious complications encountered in this series included driveline infection requiring surgical revision, septic embolus, "cleared" device infection, "suppressed" device infection, and LVAD infection treated by device removal in 1 patient and device exchange in another . CONCLUSIONS . Infection in implantable LVAD patients is common, especially in patients in whom multiple organ failure develops, requiring prolonged stay in the intensive care unit . Strategies are needed to prevent these infections in recipients of the permanent LVADs because treatment of an established infection is difficult and expensive. J Magn Reson B, 1996 Jan, 110(1), 26 - 38 Cross-correlation effects on NMR lineshapes and peptide conformation; Cuperlovic M et al.; Information about molecular structure and dynamics can potentially be obtained by studying dipole-dipole and chemical-shift anisotropy (CSA) auto-correlation and dipole-CSA cross-correlation effects in high-resolution NMR spectra . Equations for the lineshapes of the HN multiplet in the fragment- 15NH-CH- as a function of NH-CH dihedral angle are derived by including these effects within the framework of the Redfield treatment of relaxation . To test the utility of the theoretical results, 1H{15N} HSQC proton lineshape data for a variant of the enzyme staphylococcal nuclease in which all valine residues are labeled with 15N have been analyzed to obtain the conformational angle (phi) between the N-H and adjacent C-H bonds . The results are generally in good agreement with values of phi obtained from crystal structure data . Considerations in the further development of the analysis of the lineshape of the HN multiplet for experimental determinations of phi are discussed. J Bacteriol, 1996 Jan, 178(2), 531 - 6 Analysis of the Staphylococcus epidermidis genes epiF, -E, and -G involved in epidermin immunity; Peschel A et al.; The lantibiotic epidermin is produced by Staphylococcus epidermidis Tu3298 . The known genes involved in epidermin biosynthesis and regulation are organized as operons (epiABCD and epiQP) that are encoded on the 54-kb plasmid pTu32 . Here we describe the characterization of a DNA region that mediates immunity and increased epidermin production, located upstream of the structural gene epiA . The sequence of a 2.6-kb DNA fragment revealed three open reading frames, epiF, -E, and -G, which may form an operon . In the cloning host Staphylococcus carnosus, the three genes mediated an increased tolerance to epidermin, and the highest level of immunity (sevenfold) was achieved with S . carnosus carrying epiFEG and epiQ . The promoter of the first gene, epiF, responded to the activator protein EpiQ and contained a palindromic sequence similar to the EpiQ binding site of the epiA promoter, which is also activated by EpiQ . Inactivation of epiF, -E, or -G resulted in the complete loss of the immunity phenotype . An epidermin-sensitive S . epidermidis Tu3298 mutant was complemented by a DNA fragment containing all three genes . When the epiFEG genes were cloned together with plasmid pTepi14, containing the biosynthetic genes epiABCDQP, the level of epidermin production was approximately fivefold higher . The proteins EpiF, -E, and -G are similar in deduced sequence and proposed structure to the components of various ABC transporter systems . EpiF is a hydrophilic protein with conserved ATP-binding sites, while EpiE and -G have six alternating hydrophobic regions and very likely constitute the integral membrane domains . When EpiF was overproduced in S . carnosus, it was at least partially associated with the cytoplasmic membrane . A potential mechanism for how EpiFEG mediates immunity is discussed. J Bacteriol, 1996 Jan, 178(2), 462 - 9 Transcriptional regulation of the sucrase gene of Staphylococcus xylosus by the repressor ScrR; Gering M et al.; In Staphylococcus xylosus, scrB is one of two genes necessary for sucrose utilization . It encodes a sucrase that hydrolyzes intracellular sucrose-6-phosphate generated by the uptake of sucrose via the sucrose-specific enzyme II of the phosphotransferase system, the gene product of scrA . ScrB sucrase activity is inducible by the presence of sucrose in the culture medium . Primer extension experiments demonstrated that the observed regulation is achieved at the level of scrB transcription initiation . The protein mediating sucrose-specific regulation of scrB was found to be encoded immediately upstream of the sucrase gene . The nucleotide sequence of the regulatory gene scrR comprises an open reading frame that specifies a protein of 35.8 kDa . This protein exhibits similarity to transcriptional regulators of the GalR-LacI family . Inactivation of the scrR reading frame in the genome of S . xylosus led to the constitutive expression of scrB at a high level, identifying ScrR as a repressor of transcription . Sucrose-specific regulation of scrB was also lost upon deletion of 4 bp of a palindromic sequence (OB) covering positions +6 to +21 downstream of the scrB transcriptional start site . These results suggested a direct interaction of the ScrR repressor and the operator OB . Accordingly, a fusion protein consisting of the maltose-binding protein of Escherichia coli and the ScrR protein was able to interact with an scrB promoter fragment in gel mobility shift experiments but failed to bind an scrB fragment carrying the 4-bp deletion derivative of OB . An scrR promoter fragment, which dose not contain a sequence resembling OB, was not shifted by the fusion protein . This result corroborates scrR primer extension analyses showing that transcription of the repressor gene itself is not regulated . Therefore, the sucrase gene operator OB is the target sequence through which the ScrR protein exerts its negative effect on transcription initiation . In the promoter region of scrA, the gene essential for sucrose transport, two palindromic sequences that are similar to the scrB operator are found . Their presence in scrA suggests that ScrR controls a sucrose-specific regulon in S . xylosus. J Bacteriol, 1996 Jan, 178(1), 284 - 8 Serine protease EpiP from Staphylococcus epidermidis catalyzes the processing of the epidermin precursor peptide; Geissler S et al.; The function of serine protease EpiP in epidermin biosynthesis was investigated . Epidermin is synthesized as a 52-amino-acid precursor peptide, EpiA, which is posttranslationally modified and processed to the mature 22-amino-acid peptide antibiotic . epiP was expressed in Staphylococcus carnosus with xylose-regulated expression vector pCX15 . The cleavage of the unmodified EpiA precursor peptide to leader peptide and proepidermin by EpiP-containing culture filtrates of S . carnosus (pCX15epiP) was followed by reversed-phase chromatography and subsequent electrospray mass spectrometry. Nat Struct Biol, 1996 Jan, 3(1), 59 - 66 Mapping the structure of a non-native state of staphylococcal nuclease; Ermacora MR et al.; Non-native states of proteins populated at extremes of pH, or by mutation or truncation of the protein sequence, are thought to be equilibrium models for kinetic intermediates on the folding pathway . While the global physical properties of these molecules have been well characterized, analysis of their structure by NMR spectroscopy has proven difficult . Here we report the use of a new chemical cleavage technique, based on reactive oxygen species, to map the backbone fold of a truncated form of staphylococcal nuclease in a non-native state . The fragment adopts a native-like fold, however the technique also reveals regions of non-native structure. Am J Respir Crit Care Med, 1996 Jan, 153(1), 398 - 403 Surfactant subfractions during nosocomial infection in ventilated preterm human neonates; Griese M et al.; Long after resolution of the neonatal respiratory distress syndrome, deterioration of respiratory function in ventilated premature infants during severe nosocomial infections is commonly observed . Based on an increased oxygen demand and ventilatory support, impairment of the pulmonary surfactant system was hypothesized to occur . The clinical course of 10 premature neonates (764 +/- 57 g, 26.6 +/- 0.4 wk) with nosocomial infection mainly due to Staphylococcus epidermidis was divided into four periods in each individual patient: "before deterioration" (average 8 to 11 d of life), "deterioration" (11 to 17 d), "peak" (17 to 22 d), and "recovery" (22 to 24 d) . A total of 810 airway specimens were obtained by small volume lavage (1 ml/kg bw), pooled to yield appropriate amounts for differential centrifugation into two distinct subfractions known as large surfactant aggregates (LA) and small surfactant aggregates (SA) . "Before deterioration" the amount of phospholipids recovered was constant, and the two fractions were characterized by electron microscopic morphology and biochemical analysis . In the LA fraction lamellar body-like lipid structures were demonstrated, and the phospholipid composition was typical of pulmonary surfactant in premature neonates with a high content of phosphatidylcholine and phosphatidylinositol . With "deterioration" and "peak" the masses of total phospholipids and of phosphatidylcholine recovered were reduced (p < 0.05) . At the same time the mass ratio of SA/LA for phosphatidylcholine decreased from 0.32 +/- 0.10 to 0.18 +/- 0.03, indicating a more pronounced decrease of the SA fraction (p < 0.05) . The phospholipid composition in the LA fraction did not change during the course of nosocomial infection . In the SA fraction a decrease of phosphatidylcholine and a concomitant increase in lysophosphatidylcholine were observed at the "peak" of the infection . We concluded that, in ventilated premature neonates during nosocomial infection and respiratory deterioration, changes in phospholipid subfractions occur, possibly indicating impairment of pulmonary surfactant metabolism . These findings may be important when considering treatment of acute lung injury with nebulized exogenous surfactant. J Infect Dis, 1996 Jan, 173(1), 212 - 8 Intracellular pathways involved in tumor necrosis factor-alpha release by human monocytes on stimulation with lipopolysaccharide or staphylococcal peptidoglycan are partly similar; Mattsson E et al.; This study compared the effects of intracellular pathway inhibitors on tumor necrosis factor-alpha (TNF-alpha) release from human monocytes . Cells were stimulated with peptidoglycan (PG) from Staphylococcus epidermidis or with Escherichia coli lipopolysaccharide (LPS), both in the presence of 10% human serum . Of 10 substances tested, only the protein tyrosine kinase inhibitor tyrphostin AG 126 discriminated significantly between PG and LPS: TNF-alpha release induced by PG, but not by LPS, was dose-dependently suppressed . The results obtained with other modulatory substances, including different protein kinase and G protein inhibitors, suggest that calmodulin-dependent protein kinase, protein tyrosine kinase, and a cholera-toxin-sensitive G protein are involved in both PG- and LPS-induced TNF-alpha release . Further, drugs such as pentoxifylline, chloroquine, and the antioxidant apocynin similarly inhibited TNF-alpha release by PG- as well as LPS-stimulated cells. J Urol, 1996 Jan, 155(1), 155 - 7 The prosthesis salvage operation: immediate replacement of the infected penile prosthesis; Brant MD et al.; PURPOSE: We describe our experience with salvage of the infected penile prosthesis at initial presentation in 11 patients . MATERIALS AND METHODS: All patients with prosthesis infection who presented since 1991 were considered for salvage surgery . Contraindications to a salvage operation included necrotic infections, diabetic patients with purulence in the corporeal bodies, rapidly developing infections and erosion of the device cylinders . RESULTS: In 1 patient in this group a salvage attempt was repeated after re-infection, for an overall success rate of 91% . Mean followup for the group was 21 months (range 9 to 42) . Staphylococcus epidermidis was the infecting organism in 75% of our patients . CONCLUSIONS: Our experience demonstrates the safety and advantages of the immediate salvage technique. Ann N Y Acad Sci, 1995 Dec 29, 771, 472 - 84 Substance P and stress-induced changes in macrophages; Chancellor-Freeland C et al.; The present paper further links nervous-endocrine-immune systems by describing influences of SP on the immune system, and more specifically, on macrophage function . We have discussed how macrophages are important to immune responses in that much of cellular and humoral responses depend on macrophage function . Macrophages are sensitive to stress in that cold-water stress causes increased cytokine production, either spontaneously (IL-1), or after induction with LPS (IL-6, TNF alpha) . Increased cytokine levels (IL-1, IL-6) may induce acute phase reactants in the liver, which is presumably the mechanism operative in the studies indicating increases in acute phase reactants after certain stressors in animals . SP is a likely candidate to affect immune function . Previous data show that macrophages from various species have receptors for and respond to SP in vitro . SP stimulates phagocytic and chemotactic capacity, as well as increased cytokine, PGE2, and thromboxane B2 production . SP is also involved in neurogenic inflammation and is likely to be involved in the pathogenesis of several inflammatory diseases . Present data indicate SP's involvement in macrophage responses to stress . We have shown that stress induced differential SP receptor binding to peritoneal macrophages, although the precise nature of binding differences has not yet been clearly elucidated . Stress also induces more immunoreactive SP in the peritoneal fluid that bathes the peritoneal macrophages . We hypothesize that the two events, altered SP binding and concomitant increased ligand, are causally related . In addition to other correlational data showing concomitant increased SP binding plus ligand concentrations, there is more direct evidence that SP ligand may induce SP receptor expression since the SP antagonist, CP-96,345, prevents the induction of SP receptor mRNA in the staphylococcal toxin A-induced gastroenteritis (C . Pothoulakis and S . E . Leeman, personal communication) . Further supporting our notion for a causal relationship we have found the elimination of SP in vivo (via capsaicin pretreatment) reduced SP binding, as has been previously reported . We have also examined the role of SP on stress-induced altered macrophage function in vitro . SP greatly enhanced the LPS-induced macrophage TNF alpha production from stressed animals; in contrast, it produced relatively little effect on macrophages from control animals . Capsaicin pretreatment diminished the enhanced cytokine production in response to stress, such that levels of TNF alpha and IL-6 approximated those of control mice . Taken together, past and present data suggest that (1) stress may initiate, or at least contribute to, an inflammatory response, and that (2) SP is involved in the macrophage stress response . SP has long been known to be involved in inflammatory processes; our data further suggest its role in mediating stress-induced cytokine alterations. J Biol Chem, 1995 Dec 29, 270(52), 31008 - 15 Purification, characterization, and cDNA cloning of a 27-kDa lectin (L10) from horseshoe crab hemocytes; Okino N et al.; We separated granular components of horseshoe crab hemocytes by a combination of centrifugation on sucrose density gradient and high performance liquid chromatography, and a 27-kDa protein termed L10 was found to be a major component in the large granules (Shigenaga, T., Takayenoki, Y., Kawasaki, S., Seki, N., Muta, T., Toh, Y., Ito, A., and Iwanaga, S . (1993) J . Biochem . (Tokyo) 114, 307-316) . In the present work, lectin activity of this protein and its unique primary structure were elucidated . L10 was purified by four steps of chromatography, including dextran sulfate-Sepharose CL-6B, CM-Sepharose CL-6B, Sephacryl S-200, and Mono S . At least three 27-kDa isoproteins, named L10a, L10b, and L10c, were isolated . Their amino acid compositions were almost indistinguishable, and there were no amino sugars . All the isoforms had hemagglutinating activity against human A-type erythrocytes, in a Ca(2+)-independent manner with L10b showing the highest activity . The L10b-mediated hemagglutination was inhibited in the presence of N-acetylglucosamine or N-acetylallolactosamine, and the association constant (Ka) between L10b and N-acetylglucosamine was 1.95 x 10(4) M-1 . Furthermore, L10b specifically agglutinated Staphylococcus saprophyticus KD . Ultracentrifugation analysis revealed that L10b is present in monomer form in solution . A cDNA coding for an isoform of L10 was isolated from a hemocyte cDNA library . The open reading frame of the 768-base pair cDNA coded for the signal sequence of 19 residues . The mature protein had 236 residues with the calculated molecular weight of 26,757 . Amino acid sequences of the peptides derived from L10c exactly corresponded to the predicted sequence of the cDNA, whereas amino acid replacements of Ile-129 to Val and His-213 to Tyr existed both in L10a and L10b, suggesting that the cDNA codes for L10c . Cysteine was absent and there were five tandem repeats with 47 amino acids in each segment with internal sequence identities of 49-68% . The entire amino acid sequences had no significant sequence similarity with other known proteins. Proc Natl Acad Sci U S A, 1995 Dec 19, 92(26), 12156 - 9 Isolation of HLA-DR1.(staphylococcal enterotoxin A)2 trimers in solution; Tiedemann RE et al.; Mutational studies indicate that the superantigen staphylococcal enterotoxin A (SEA) has two separate binding sites for major histocompatibility complex (MHC) class II molecules . Direct evidence is provided here for the formation of SEA-MHC class II trimers in solution . Isoelectric focusing separated SEA-HLA-DR1 complexes into both dimers and HLA-DR1.SEA2 trimers . The molar ratio of components was determined by dual isotope labeling . The SEA mutant SEA-F47S, L48S, Y92A, which is deficient in MHC class II alpha-chain binding, formed only dimers with HLA-DR1, whereas a second SEA mutant, SEA-H225A, which lacks high-affinity MHC class II beta-chain binding was incapable of forming any complexes . Thus SEA binding to its MHC receptor is a two-step process involving initial beta-chain binding followed by cooperative binding of a second SEA molecule to the class II alpha chain. Thromb Res, 1995 Dec 15, 80(6), 509 - 18 Sphingomyelinase and cell-permeable ceramide analogs increase the release of plasminogen activator inhibitor-1 from cultured endothelial cells; Soeda S et al.; We investigated the effect of exogenous staphylococcal sphingomyelinase (SMase) on the release of tissue-type plasminogen activator (t-PA) and plasminogen activator inhibitor-1 (PAI-1) from cultured human umbilical vein endothelial cells (HUVEC) . Addition of SMase (2 units/ml) to the culture medium induced an approx . 15-fold increase in the extracellular level of PAI-1 antigen at 3 h . No significant increase in the level of t-PA antigen was detected . Treatment of HUVEC with SMASE (2 units/ml) for 3 h resulted in a significant decrease in the cellular sphingomyelin (SM) level, accompanied by a corresponding increase in the ceramide level . Cell-permeable ceramide analogs also enhanced the release of PAI-1 from cultured HUVEC in concentration- and time-dependent manners . A 6-fold increase in PAI-1 antigen level was observed after incubation for 3 h with 10 microM N-acetylsphingosine . Similar effect was noted as early as 2 h with 10 microM N-hexyanoylsphingosine . Addition of sphingosine failed to affect the release of PAI-1 from cultured HUVEC, indicating that the effects of ceramide analogs were independent of sphingosine generation . Pretreatment with cycloheximide or actinomycin D abated the response of HUVEC to N-acetylsphingosine in the increased levels of both extracellular and intracellular PAI-1 . These results suggest that ceramide, generated via "SM cycle", acts as a lipid mediator of PAI-1 release from vascular endothelial cells, and may contribute to a better understanding of the pathogenesis of the PAI-1-associated thrombotic disorders. FEMS Microbiol Lett, 1995 Dec 15, 134(2-3), 165 - 9 Functional cloning of the dihydropteroate synthase gene of Staphylococcus haemolyticus; Kellam P et al.; A 1,7-kilobase fragment of the Staphylococcus haemolyticus chromosome containing the dihydropteroate synthase gene has been cloned by complementation in a temperature-sensitive mutant of Escherichia coli . The gene, designated folP, predicts a gene product of 29613 Da which shares significant amino acid sequence identity with other known bacterial dihydropteroate synthases . Analysis of the DNA sequence upstream and downstream of folP identified two further, incomplete open reading frames, one of which shows predicted amino acid sequence similarity to a second bacterial folic acid synthesis enzyme, dihydroneopterin aldolase. Cancer Res, 1995 Dec 15, 55(24), 6140 - 5 Analysis of the T cell receptor in the lymphoproliferative disease of granular lymphocytes: superantigen activation of clonal CD3+ granular lymphocytes; Zambello R et al.; To investigate whether cell populations in CD3+ lymphoproliferative disease of granular lymphocytes (LDGLs) were skewed toward the use of specific V beta regions, we studied the repertoire of T-cell receptor (TCR) V beta gene products in 18 patients, as well as their relationship to the clonal bands in the Southern blot and the activation mediated by superantigens . Using a panel of monoclonal antibodies (mAbs) for conserved V beta segments and PCR, a dominant population expressing a specific V beta region was demonstrated in all patients . In five (27%) cases, granular lymphocytes (GLs) were found to express the V beta 13.1, while V beta 8 and V beta 6 were each expressed in three (17%) cases . The remaining cases were characterized by the proliferation of TCR V beta 2, V beta 3, V beta 4, V beta 9, V beta 12, V beta 16, and V beta 20 . This finding indicates a biased usage of a limited TCR V beta in LDGLs, since nearly 60% of the cases utilized only three families of the TCR V beta genes . In all of the cases studied, we proved that the subset recognized by mAb and PCR was identical to that accounting for the extra band(s) of the Southern blot . This finding confirms the clonal nature of the population identified according to TCR V beta expression both by phenotype and PCR . On functional grounds, we evaluated whether GLs can be activated through the specific TCR using the superantigens recognizing discrete V beta families, such as staphylococcal proteins, including SEA, SEB, SEC1, SEC2, SED, and SEE . We demonstrated that the TCR-alpha/beta of clonal GLs in LDGL patients is functionally active in delivering cytotoxic and proliferative signals upon superantigen activation. Biochemistry, 1995 Dec 12, 34(49), 15895 - 905 The equilibrium folding pathway of staphylococcal nuclease: identification of the most stable chain-chain interactions by NMR and CD spectroscopy; Wang Y et al.; In a previous report {Alexandrescu, A . T., Abeygunawardana, C., & Shortle, D . (1994) Biochemistry 33, 1063-1072}, NMR methods were used to characterize the residual structure in delta 131 delta, a large fragment of staphylococcal nuclease that serves as a model denatured state under nondenaturing conditions . On the basis of a large number of missing amide protons for the residues that form a three-strand antiparallel beta sheet in the native state, it was concluded that this beta meander may be highly populated in delta 131 delta, with severe line broadening due to relatively slow exchange between different conformational states . In the present report, results from circular dichroism spectroscopy and NMR spectroscopy indicate strands beta 2-beta 3 form a beta hairpin at urea concentrations below 6 M . Amide proton resonances from several hydrophobic residues adjacent to this beta hairpin disappear in concert with all of the beta 2-beta 3 residues, suggesting a local, non-native hydrophobic interaction may help stabilize the beta hairpin . At concentrations below 3 M, all amide resonances from strand beta 1 in delta 131 delta also disappear, suggesting that beta 1 may combine with the beta 2-beta 3 hairpin to form a native-like beta meander . In addition, the hydrophobic helix alpha 2 decreases from approximately 30% population in 0 M urea to approximately 10%-15% at 6 M urea, whereas helix alpha 1 goes from 10%-15% populated in 0 M urea to undetectable in 6 M urea . Characterization of a second, distinctly different denatured state, WT nuclease at pH 3.0 and low salt, reveals that this low-density acid-denatured state is structurally similar to delta 131 delta at low concentrations of urea . From these and previously published data, a tenative equilibrium folding pathway can be constructed for staphylococcal nuclease which describes the relative strengths and interdependencies of the chain-chain interactions involved in forming the native state. Biochem Biophys Res Commun, 1995 Dec 5, 217(1), 74 - 80 IL-12 rescues galactosamine-loaded mice from lethal shock triggered by staphylococcal enterotoxin; Takahashi I et al.; Staphylococcal enterotoxin B (SEB) causes lethal shock in D-galactosamine sensitized mice . The lethal shock triggered by SEB is mediated by T cells . We found that the lethal shock was restricted by MHC class II molecule . In addition, consecutive oral exposures to SEB induced tolerance against the shock in the SEB-sensitive mice . To elucidate the tolerance mechanism, the role of anti-inflammatory cytokines was examined . RT-PCR analysis revealed that CD4+ T cells from the SEB-sensitive mice expressed significant levels of IFN-gamma, IL-2, IL-4 and IL-10 mRNA, while those from the tolerant mice exhibited significant levels of IFN-gamma but not IL-2 or IL-4 mRNA . These results indicate that polarity of T helper (Th) cells from Th0 to Th1 was involved in the tolerance to the SEB-induced lethal shock . Lymphoid tissues of the tolerant mice generated mRNA of IL-12, a cytokine which favors Th1 response . It was also demonstrated that intraperitoneal administration of IL-12 conferred protection against the lethal shock in the sensitive mice. Biochemistry, 1995 Dec 5, 34(48), 15700 - 3 Influence of phosphoenolpyruvate and magnesium ions on the quaternary structure of enzyme I of the phosphotransferase system from gram-positive bacteria; Hubner G et al.; Solution X-ray scattering patterns of enzyme I of the phosphotransferase system from Staphylococcus carnosus indicate an increase in radius of gyration and molecular mass in the presence of Mg2+ or both Mg2+ and phosphoenolpyruvate, indicating a partial dimerization of enzyme I . Mg2+ ions are essential for both the dimerization and the activation, whereas the substrate phosphoenolpyruvate shifts the monomer--dimer equilibrium to the enzymatically active dimer by decreasing the dissociation rate of the phosphorylated dimer. Ned Tijdschr Geneeskd, 1995 Dec 2, 139(48), 2498 - 501 {A patient with acute leukemia and meningitis caused by Staphylococcus epidermidis treated with fosfomycin}; Silbermann MH et al.; In a 17-year-old male patient with acute lymphoblastic leukaemia, who was being treated with chemotherapy, a Staphylococcus epidermidis infection with several septicaemias developed during a period of protracted neutropenia . The patient was treated with vancomycin and fusidic acid, but blood cultures remained positive . The patient also developed staphylococcal meningitis . After the antibiotic regimen was supplemented by fosfomycin, the blood cultures became sterile . Combination treatment with vancomycin and fosfomycin was continued for two months without apparent toxicity . In individual cases of infection with multiresistant S . epidermidis fosfomycin may be included in the antibiotic regimen . This is the first report of parenteral use of fosfomycin in the Netherlands. Ophthalmology, 1995 Dec, 102(12), 1925 - 31 Endophthalmitis after secondary intraocular lens implantation . A case-report study; Scott IU et al.; PURPOSE: To investigate whether endophthalmitis is more common after secondary intraocular lens (IOL) implantation than after other intraocular surgeries . Risk factors, causative organisms, and visual acuity outcomes of endophthalmitis after secondary IOL implantation were investigated . METHODS: The ten cases consisted of all patients treated at the Bascom Palmer Eye Institute between January 1, 1984, and December 31, 1994, for endophthalmitis after secondary IOL implantation . The 34 control patients consisted of all patients who underwent secondary IOL implantation within 3 years of the case patients (6-year span) by the same surgeons but who did not develop endophthalmitis . Demographic and clinical data were abstracted from patients' medical records . RESULTS: Fifty percent (5 of 10) of case patients had a history of diabetes mellitus, compared with 5.9% (2 of 34) of control patients . All three posterior chamber IOLs among case patients had transscleral suture fixation, whereas all four posterior chamber IOLs among control patients were in the sulcus without scleral sutures . Case patients were more likely than control patients to have IOLs with polypropylene haptics (30.0% and 5.9%, respectively) and to have had their IOLs placed through a superior rather than a temporal incision (70.0% and 17.9%, respectively) . Eighty percent of case patients and 14.7% of control patients had either eyelid abnormalities (marked blepharitis, ectropion) or postoperative wound defects . Staphylococcus epidermidis was isolated in 50% of case patients . After treatment, 90.0% of case patients achieved a visual acuity of greater than or equal to 20/100 . CONCLUSION: Endophthalmitis after secondary IOL implantation was associated with diabetes, transscleral suture fixation of posterior chamber IOLs, polypropylene haptics, preoperative eyelid abnormalities, re-entry of eye through a previous wound, and postoperative wound defects . Most cases were caused by Staphylococcus epidermidis, and the post-treatment visual outcomes were generally good. J Hematother, 1995 Dec, 4(6), 571 - 7 Bifunctional antibody retargeting in vivo-activated T lymphocytes: simplifying clinical application; Chapoval AI et al.; For antitumor x anti-CD3 bifunctional antibody (BFA) therapy to be clinically relevant in solid tumors, activated lymphocytes must be present within tumors . Toward that end, three uniquely different in vivo activation approaches were investigated in a p97 human antigen expressing syngeneic murine melanoma model . beta-Glucan (200 micrograms), staphylococcal enterotoxin B (SEB) (50 micrograms), and F(ab')2 BFA (10 micrograms) were tested for their ability to activate lymphocytes, neutralize pulmonary metastases, and treat established tumors . Systemic activation, measured as the ability of splenocytes to lyse tumor cells in vitro in the presence of BFA, was enhanced by the in vivo administration of SEB but not by beta-glucan or F(ab')2 BFA . Despite lacking a systemic effect, F(ab')2 BFA increased both direct and BFA-mediated cytotoxicity in fresh tumor infiltrating lymphocytes . beta-Glucan did not increase systemic or intratumor T cell activation . However, it significantly enhanced the ability of splenocytes to lyse NK-sensitive YAC-1 cells . When tested in a pulmonary metastases model, all three forms of immune modulation combined with F(ab')2 BFA significantly reduced the number of metastases . BFA were more effective at tumor neutralization when combined with SEB compared with adoptively transferred, in vitro-activated splenocytes . These studies demonstrate that immune modulators when combined with F(ab')2 BFA can provide effective antitumor therapy . Several clinical obstacles may be overcome by the application of these reagents. J Arthroplasty, 1995 Dec, 10(6), 817 - 22 Bacterial growth on antibiotic-loaded acrylic cement . A prospective in vivo retrieval study; Kendall RW et al.; Twenty-three patients with intraoperative culture-proven periprosthetic infection of the hip or knee were enrolled in a prospective cement retrieval study . All were treated with a two-stage technique using antibiotic-loaded acrylic cement as an antibiotic depot . Staphylococcus epidermidis was the most commonly isolated organism (19 of 23 cases) . Cement and tissue were examined at second-stage revision for the presence of viable organisms . In this series, no organisms were isolated from the surface of the cement, a 100% concordance with the tissue cultures . A subsequent failure rate of 4.4% (1 case) was seen in this series . Investigation suggests this may represent reinfection from a new strain of organism rather than failure of eradication of the original infection. J Arthroplasty, 1995 Dec, 10(6), 716 - 21 Postoperative infection following orthopaedic surgery in human immunodeficiency virus-infected hemophiliacs with CD4 counts < or = 200/mm3; Ragni MV et al.; Human immunodeficiency virus-infected hemophiliacs are at risk for bacterial and opportunistic infections with worsening immunosuppression . Thus, the risk of postoperative infection following orthopaedic surgery is of considerable concern . A survey of United States hemophilia treatment centers was conducted to determine the incidence of postoperative infection in human immunodeficiency virus-positive hemophiliacs with CD4 counts of 200 mm3 or less undergoing orthopaedic surgery . A total of 115 centers from 37 states reported that postoperative infection occurred in 10 (15.1%) of 66 patients undergoing 74 orthopaedic procedures, between several weeks and 5 months following surgery . In five (50%), pre-operative infection preceded postoperative joint infection . Staphylococcus was the most common organism isolated in a prosthetic joint infection, in 6 of 10 (60.0%), and the knee was the most commonly affected joint, in 9 of 10 (90.0%) . Joint arthroplasty appeared to have 10 times the risk of nonarthroplasty procedures for postoperative infection (9 of 34 {26.5%} and 1 of 40 {2.5%}, respectively, P < .01) . Two subjects developed chronic osteomyelitis . The rate of postoperative infection in human immunodeficiency virus-positive hemophiliacs with CD4 counts of 200/mm3 or less appears to be high, when compared with the general population . Early, vigorous treatment should be instituted for suspected infection, antibiotic prophylaxis considered for invasive procedures, and surgical intervention individualized based on the balance of risks and benefits. East Afr Med J, 1995 Dec, 72(12), 800 - 4 Quality of intravenous infusion fluids manufactured in Kenya; Aluoch-Orwa JA et al.; The incidence and nature of microbial contamination of intravenous fluids prepared by four manufacturing establishments in Kenya was evaluated using the European Pharmacopoeia membrane filtration method for sterility testing . The percentage failures were 28.6% for source D, 18.8% for source A, 12.5% for source B and 10.5% for source C . The major contaminant was aspergillus which was isolated from samples from three sources . Candida and Staphylococcus accounted for the contamination of samples from two sources . Failure rates due to the chemical composition of the products was 66.7% for Source A, 60.0% for D, 41.7% for C and 13.3% for B . The experience of the manufacturing sites appeared to correlate with the quality of the products, with the older manufacturing establishments showing lower percentage failures. Eur J Clin Microbiol Infect Dis, 1995 Dec, 14(12), 1052 - 6 A simple and inexpensive method for the identification of Staphylococcus epidermidis and Staphylococcus hominis; Mulder JG; Eight hundred and ninety-two strains of Staphylococcus species were identified by means of desferrioxamine susceptibility and fermentation results of three carbohydrates, with the API Staph system (bioMerieux, France) as reference method . No identification could be obtained for 34 strains with API Staph . Of the remaining 858 strains, identical identification was obtained with 842 (98.1%) . All 707 strains identified as Staphylococcus epidermidis or Staphylococcus hominis by the API Staph system were found to be desferrioxamine susceptible, and all but 5 (3.3%) of 151 strains identified as other staphylococcal species were found to be resistant, yielding an identification correlation of 99.4% for desferrioxamine . The five additional strains which were susceptible to desferrioxamine were identified as Staphylococcus capitis (2 strains), Staphylococcus lugdunensis (2 strains), and Staphylococcus warneri (1 strain) by API Staph, and as Staphylococcus epidermidis (1 strain), Staphylococcus hominis (3 strains), and one other staphylococcal species by the experimental system. AIDS Res Hum Retroviruses, 1995 Dec, 11(12), 1433 - 9 HLA class II on HIV particles is functional in superantigen presentation to human T cells: implications for HIV pathogenesis; Rossio JL et al.; The mechanisms of immune suppression by the human immunodeficiency virus, HIV-1, are more complex than simple helper T cell deletion via infection and viral-induced lysis . Since the recent description of cellular proteins associated with HIV suggests that these proteins may be active in viral pathogenesis, the nature of HLA class II gene product carried on HIV, one of the most abundant of the human components carried with the virus, was examined . HIV bearing HLA-DR was shown to act with bacterial superantigen, staphylococcal enterotoxin A (SEA), to stimulate highly purified human T lymphocytes . T cell stimulation by wild-type HIV was shown by both induction of proliferation and by production of the cytokine interleukin 2 (IL-2) . In contrast, HIV produced from mutant cells lacking class II genes were unable to cooperate with SEA to activate T cells . Neither whole HIV nor several proteins purified from HIV (gp120, gp41, p24, p7, and p6) exhibited superantigen-like activity in this system . HLA-DR-bearing HIV could, in the continued presence of SEA, induce T cell apoptosis, as detected by nuclear fragmentation and morphological criteria . These data indicate that human cellular proteins associated with HIV may be biologically active, and these proteins should be considered in mechanisms of viral pathogenicity and immunogenicity. Protein Sci, 1995 Dec, 4(12), 2545 - 58 Chemically crosslinked protein dimers: stability and denaturation effects; Byrne MP et al.; Nine single substitution cysteine mutants of staphylococcal nuclease (nuclease) were preferentially crosslinked at the introduced cysteine residues using three different bifunctional crosslinking reagents; 1,6-bismaleimidohexane (BMH), 1,3-dibromo-2-propanol (DBP), and the chemical warfare agent, mustard gas (bis(2-chloroethyl)sulfide; mustard) . BMH and mustard gas are highly specific reagents for cysteine residues, whereas DBP is not as specific . Guanidine hydrochloride (GuHCl) denaturations of the resulting dimeric proteins exhibited biphasic unfolding behavior that did not fit the two-state model of unfolding . The monofunctional reagent, epsilon-maleimidocaproic acid (MCA), was used as a control for the effects of alkylation . Proteins modified with MCA unfolded normally, showing that this unusual unfolding behavior is due to crosslinking . The data obtained from these crosslinked dimers was fitted to a three-state thermodynamic model of two successive transitions in which the individual subunits cooperatively unfold . These two unfolding transitions were very different from the unfolding of the monomeric protein . These differences in unfolding behavior can be attributed in large part to changes in the denatured state . In addition to GuHCl titrations, the crosslinked dimers were also thermally unfolded . In contrast to the GuHCl denaturations, analysis of this data fit a two-state model well, but with greatly elevated van't Hoff enthalpies in many cases . However, clear correlations between the thermal and GuHCl denaturations exist, and the differences in thermal unfolding can be rationalized by postulating interactions of the denatured crosslinked proteins. Eur J Immunol, 1995 Dec, 25(12), 3437 - 44 Definition of sites on HLA-DR1 involved in the T cell response to staphylococcal enterotoxins E and C2; Hargreaves RE et al.; We have exploited the relative inefficiency of interaction between staphylococcal enterotoxins, SEE or SEC2, and H-2Ek compared to HLA-DR1 molecules to deduce which regions of the major histocompatibility complex (MHC) class II molecule are involved in the T cell response to these superantigens . Transfectants expressing hybrid DR/H-2E MHC class II molecules were used to present SEE to the T cell receptor V beta 8.1-expressing Jurkat cell line, and SEC2 to human peripheral blood T cells . For SEE, the critical region of the class II molecule for T cell reactivity and for binding was the beta 1 domain alpha-helix . The functional data were corroborated by measurements of direct binding . Sequence comparison between DR and H-2E raised the possibility that the glutamic acid at position 84 in the beta chain of H-2Ek, in place of glycine was responsible for the observed functional effects . This suggestion was supported by the finding that DQw2 (glutamine at 84) transfectants supported the SEE response much more efficiently than DQw6 that has glutamic acid at this position . In addition, amino acid substitutions at either position 36 or 39 in the DR alpha 1 domain abolished T cell reactivity without any obvious alteration in binding . For SEC2, use of transfectants expressing exon-shuffled alpha and beta chain genes showed that replacement of the alpha 1, alpha 2 and beta 1 domains with H-2E sequence inhibited the presentation of SEC2 . Similarly, the substitutions at positions 36 and 39 in the alpha 1 domain abolished the T cell response to SEC2 . Taken together, these data may be best explained by a model in which these two toxins have primary binding sites on the beta 1 domain (SEE) and the alpha 1 and alpha 2 domains (SEC2), but by virtue of a secondary binding site on the opposite surface of the class II molecule, cross-link two adjacent DR molecules . Such cross-linking may be important in the induction of T cell reactivity. Eur J Immunol, 1995 Dec, 25(12), 3425 - 30 Bacterial superantigen specificities of mouse T cell receptor V beta 20; Bravo de Alba Y et al.; The study of the mouse T cell receptor (TcR) beta chain repertoire in BALB/c thymocytes led to the identification of the V beta 20 gene segment . The expression of V beta 20 estimated at the transcriptional level differs among mouse strains, suggesting clonal deletion . In the present study, we reconstituted by transfection functional TcR using the V beta 20 segment with different V alpha segments and studied the action of superantigen toxins . The V beta 20-transfectant T cells are activated by staphylococcal enterotoxins A and E (SEA and SEE) but not by the other tested toxins . The activation is dependent on the presence of cells expressing major histocompatibility complex class II molecules . Different HLA DR alleles can present the bacterial toxins, establishing that they interact with TcRV beta 20 as superantigens . Moreover, the V alpha domain associated with the V beta 20 domain has an influence on the response to these toxins . The fact that V beta 20 is recognized by SEA and SEE, although both toxins are known to interact with different sets of V beta, suggests the presence of different TcR binding sites on the toxins. J Immunol, 1995 Dec 1, 155(11), 5220 - 6 Regulation of JAK3 expression and activation in human B cells and B cell malignancies; Tortolani PJ et al.; Members of the Janus family (JAK) of protein tyrosine kinases are critical enzymes in signaling pathways via hematopoietin receptors . We have cloned JAK3, which unlike other known family members (JAK1, JAK2, and TYK2) is preferentially expressed in hematopoietic cells but not in a variety of other cells . Functionally, JAK3 and JAK1 are coupled to the receptors for IL-2, IL-4, IL-7, IL-9, and IL-15 in T cells and NK cells . Because of the importance of IL-2, IL-4, and IL-7 in B cell physiology, we sought to determine whether JAK3 was also present in B lymphocytes and whether it was involved in signaling via cytokines that are important for B cell development and function . In this report, we demonstrate that JAK3 is expressed in normal human peripheral blood B cells at levels that are comparable to those in T cells . In addition, the levels were found to be markedly up-regulated following stimulation with staphylococcal protein A Cowan and anti-CD40 Abs . In addition, IL-4 and IL-7 induced the rapid tyrosine phosphorylation of JAK3 and JAK1, and IL-4 activated both JAK3 and JAK1 phosphotransferase activity . JAK3 protein was also detected in immature B cell lines, but not in more well differentiated cell lines . Additionally, JAK3 was detected in lysates from bone marrow lymphoblasts of patients with B cell precursor acute lymphocytic leukemia and cell lines derived from human B cell lymphomas . Together, these data suggest that the regulation of JAK3 expression and activity is likely to be important in B cell development and function. J Immunol, 1995 Dec 1, 155(11), 5083 - 7 Defective TCR-mediated signaling in anergic T cells; Migita K et al.; Extrathymic T cell tolerance to Ags can be achieved through clonal deletion by activation-induced programmed cell death as well as by functional unresponsiveness (anergy) of Ag reactive-T cells . Previous studies demonstrated that in vivo administration of staphylococcal enterotoxin B (SEB) induces SEB-specific T cell anergy . To investigate the molecular mechanisms of T cell anergy, we analyzed the TCR-mediated signal transduction in normal and anergic T cells . Anergic T cells exhibited impaired protein tyrosine phosphorylation on TCR-mediated stimulation . The altered tyrosine phosphorylation in anergic T cells may be caused by a defect in tyrosine phosphorylation of TCR zeta-chain and its subsequent association with ZAP-70 tyrosine kinase . Our data demonstrated that TCR zeta-chain phosphorylation and its sequential interaction with ZAP-70 are essential for the initiation of TCR-mediated signal transduction. Clin Immunol Immunopathol, 1995 Dec, 77(3), 332 - 8 Modulatory effects of staphylococcal superantigen TSST-1 on IgE synthesis in atopic dermatitis; Lester MR et al.; Atopic dermatitis (AD) is a chronic skin disorder associated with elevated serum IgE and colonization of the skin by Staphylococi which secrete toxins with superantigenic activity . The present study examined the immunomodulatory effects of toxic shock syndrome toxin (TSST)-1, a prototypic superantigen, on IgE synthesis by interleukin (IL)-4-stimulated peripheral blood mononuclear cells (PBMC) from five patients with AD and five normal subjects . TSST-1 inhibited IL-4-induced IgE synthesis by AD and normal PBMC (P < 0.05) . In contrast, IgG synthesis was not similarly affected (P > 0.30) . Inhibition of IL-4-induced IgE production was associated with induction of interferon (IFN)-gamma synthesis by TSST-1 (P < 0.02) . Normal PBMC synthesized significantly more (P < 0.005) IFN-gamma than AD PBMC . A neutralizing antibody to IFN-gamma reversed TSST-1-induced suppression of IgE synthesis by the normal PBMC (P < 0.03), but not the AD PBMC . In AD, but not normal, PBMC anti-IFN-alpha antibody reversed the suppression of IgE synthesis induced by TSST-1 . These results demonstrate that TSST-1 uses different mechanisms for modulation of IgE synthesis in AD versus normal PBMC . Furthermore, the reversal of TSST-1-induced suppression of IgE synthesis in AD PBMC by anti-IFN-alpha, but not anti-IFN-gamma, is consistent with the concept that AD is associated with defective Th1 cell function and enhanced monocyte activity. J Exp Med, 1995 Dec 1, 182(6), 1997 - 2006 Physical dissociation of the TCR-CD3 complex accompanies receptor ligation; Kishimoto H et al.; Recent studies indicate that there may be functional uncoupling of the TCR-CD3 complex and suggest that the TCR-CD3 complex is composed of two parallel signal-transducing units, one made of gamma delta epsilon chains and the other of zeta chains . To elucidate the molecular mechanisms that may explain the functional uncoupling of TCR and CD3, we have analyzed their expression by using flow cytometry as well as immunochemical means both before and after stimulation with anti-TCR-beta, anti-CD3 epsilon, anti-CD2, staphylococcal enterotoxin B, and ionomycin . We present evidence that TCR physically dissociates from CD3 after stimulation of the TCR-CD3 complex . Stimulation with anti-CD3 resulted in down-modulation of TCR within 45 min whereas CD3 epsilon was still expressed on the cell surface as detected by flow cytometry . However, the cell surface expression of TCR and CD3 was not affected when cells were stimulated with anti-TCR-beta under the same conditions . In the case of anti-CD3 treatment of T cells, the TCR down-modulation appeared to be due to the internalization of TCR, as determined by immunoelectron microscopy . Immunochemical analysis of cells after stimulation with either anti-TCR or anti-CD3 mAbs revealed that the overall protein levels of TCR and CD3 were similar . More interestingly, the dissociation of the TCR-CD3 complex was observed with both treatments and occurred in a manner that the TCR and the associated TCR-zeta chain dissociated as a unit from CD3 . These results provide the first report of physical dissociation of TCR and CD3 after stimulation through the TCR-CD3 complex . The results also suggest that the signal transduction pathway triggered by TCR may differ from that induced by CD3. Cell Immunol, 1995 Dec, 166(2), 227 - 35 Superantigen reactivity of gamma delta T cell clones isolated from patients with multiple sclerosis and controls; Stinissen P et al.; gamma delta T cells have been implicated as playing a role in the demyelinating processes of multiple sclerosis (MS) . However, the nature of the ligands which lead to activation and accumulation of gamma delta T cells in the brain lesions remains unknown . This study was undertaken to examine whether gamma delta T cells derived from cerebrospinal fluid and blood of MS patients could be stimulated by bacterial superantigens: staphylococcal enterotoxins A and B and toxic shock syndrome toxin-1 . A panel of 16 gamma delta T cell clones isolated from MS patients and controls was found to react with the superantigens used at a nanogram range and displayed specific cytotoxic activity toward target cells pulsed with the corresponding superantigens . The responses of the gamma delta T cell clones did not require MHC-matched accessory cells and were not blocked by antibodies to the MHC molecules, suggesting a non-MHC restricted interaction . The superantigen reactivity was associated with both V delta 2+/V gamma 2+ and V delta 1+/V gamma 1+ subsets, reportedly found in the MS lesions . Our data suggest an alternative pathway which may account for gamma delta T cell activation in MS. Cell Immunol, 1995 Dec, 166(2), 187 - 95 Antigen-induced death of alloreactive human T-lymphocytes occurs in the absence of low molecular weight DNA fragmentation; Pohl T et al.; Stimulation via the CD3/TCR molecular complex induces proliferation of resting T cells, but triggers programmed cell death (apoptosis) in immature thymocytes and preactivated mature T cells . Activation-induced cell death (AICD) triggered by anti-CD3/TCR mAb or by staphylococcus enterotoxin superantigen is associated with fragmentation of genomic DNA into oligonucleosomal fragments of 200 bp length, thus displaying the characteristic features of apoptosis . Here, we show that a fraction (20-50%) of cells in alloreactive CD8 human short-term T cell lines, generated by repeated restimulation with EBV-transformed B cell lines, undergo AICD when restimulated with the appropriate (but not with third party) stimulator cells . AICD of responder T cells is inhibited when stimulator cells are preincubated with anti-HLA class I mAb but not with anti-HLA class II mAb, indicating that T cell death is dependent on alloantigen (HLA class I) recognition by responding CD8 T cells . Importantly, alloantigen-induced T cell death occurs in the absence of detectable DNA fragmentation . Thus, several independent assay systems all failed to reveal low molecular weight DNA fragmentation, even though DNA fragmentation was readily detected in T cell lines exposed to PHA or gamma-irradiation . Alloantigen-induced T cell death was prevented by aurintricarboxylic acid, which has previously been shown to inhibit apoptosis in experimental systems where no DNA fragmentation occurs . Taken together, these results demonstrate that alloantigen can trigger AICD in mature responding T cells in the absence of low molecular weight DNA fragmentation. J Mol Biol, 1995 Dec 1, 254(3), 381 - 91 Determination of sequence specificity between a plasmid replication initiator protein and the origin of replication; Thomas CD et al.; Staphylococcal plasmids of the pT181 family replicate by a rolling circle mechanism, requiring the activities of a plasmid-specified Rep protein . The initiation event involves site-specific phosphodiester bond cleavage by Rep within the replication origin, ori . In vitro the Rep proteins also display type-I topoisomerase activity specific for this plasmid family . Although the single site of bond cleavage, ICR II, is conserved among all members of the pT181 family, the plasmid-specific Rep proteins are able to discriminate between family members in vivo, initiating replication only from the cognate origin . The basis of such specificity is believed to be due to a non-covalent binding interaction between Rep and a DNA sequence adjacent to the site of phosphodiester bond cleavage . Using the RepD protein specified by plasmid pC221, we present data for the physical parameters of RepD:oriD complex formation . Quantification of the relative strengths of the non-covalent interactions for different but related ori target sequences, measured by gel mobility shift experiments, has yielded data that are in accord with the known specificity of the protein in vivo . Oligonucleotide competition experiments demonstrate that this interaction is indeed attributable to the specificity determinant, ICR III . Protein-DNA crosslinking methods show that a carboxyl-terminal proteolytic fragment of RepD makes a specific interaction with the ICR III region of its cognate replication origin . Analysis of topoisomerase rates indicates that the interaction between ICR III and the carboxyl terminus of the protein is required before a productive interaction, namely the phosphodiester bond cleavage at the ICR II, can occur. Plast Reconstr Surg, 1995 Dec, 96(7), 1702 - 8 Toxic shock syndrome as a complication of breast prostheses; Poblete JV et al.; A case of a 21-year-old woman who developed toxic shock syndrome 6 days after augmentation mammaplasty with saline breast implants is reported . The infecting organism was S . aureus that was toxic shock syndrome exotoxin 1-negative and staphylococcal endotoxin B-positive . The causes and etiology of this rare postoperative complication are discussed. Proc Natl Acad Sci U S A, 1995 Nov 21, 92(24), 10995 - 9 Staphylococcal enterotoxins modulate interleukin 2 receptor expression and ligand-induced tyrosine phosphorylation of the Janus protein-tyrosine kinase 3 (Jak3) and signal transducers and activators of transcription (Stat proteins); Nielsen M et al.; Staphylococcal enterotoxins (SE) stimulate T cells expressing the appropriate variable region beta chain of (V beta) T-cell receptors and have been implicated in the pathogenesis of several autoimmune diseases . Depending on costimulatory signals, SE induce either proliferation or anergy in T cells . In addition, SE can induce an interleukin-2 (IL-2) nonresponsive state and apoptosis . Here, we show that SE induce dynamic changes in the expression of and signal transduction through the IL-2 receptor (IL-2R) beta and gamma chains (IL-2R beta and IL-2R gamma) in human antigen-specific CD4+ T-cell lines . Thus, after 4 hr of exposure to SEA and SEB, the expression of IL-2R beta was down-regulated, IL-2R gamma was slightly up-regulated, while IL-2R alpha remained largely unaffected . The changes in the composition of IL-2Rs were accompanied by inhibition of IL-2-induced tyrosine phosphorylation of the Janus protein-tyrosine kinase 3 (Jak3) and signal transducers and activators of transcription called Stat3 and Stat5 . In parallel experiments, IL-2-driven proliferation was inhibited significantly . After 16 hr of exposure to SE, the expression of IL-2R beta remained low, while that of IL2R alpha and IL2R gamma was further up-regulated, and ligand-induced tyrosine phosphorylation of Jak3 and Stat proteins was partly normalized . Yet, IL-2-driven proliferation remained profoundly inhibited, suggesting that signaling events other than Jak3/Stat activation had also been changed following SE stimulation . In conclusion, our data suggest that SE can modulate IL-2R expression and signal transduction involving the Jak/Stat pathway in CD4+ T-cell lines. J Immunol, 1995 Nov 15, 155(10), 4829 - 37 Increased susceptibility of fas mutant MRL-lpr/lpr mice to staphylococcal enterotoxin B-induced septic shock; Mountz JD et al.; MRL-lpr/lpr mice are defective in the fas Ag/APO-1 apoptosis gene (CD95) . Using the hepatotoxin D-galactosamine (D-GalNH2), we demonstrate that MRL-lpr/lpr mice have an increased susceptibility to staphylococcal enterotoxin B (SEB)-induced lethal shock, which causes them to exhibit the septic shock-like behaviors of fur ruffling and listlessness, and death occurs within 8 to 18 h . SEB susceptibility is greater in V beta 8.2 TCR transgenic MRL-lpr/lpr mice than in nontransgenic mice . In studies designed to elucidate the molecular pathways of SEB-induced septic shock, we found that C57Bl/6.Ab0/Ab0, MHC class II-deficient "C2D" mice, but not C57Bl/6-(+/+) mice, are nonresponsive to challenge with SEB . C2D mice, backcrossed with the fas mutation resulting in double-knockout C2D;lpr/lpr mice, are more susceptible to challenge with SEB/D-GalNH2 . The LD50s for C57Bl/6.C3H-gld/gld "fas ligand-mutant mice" challenged with SEB/D-GalNH2 were comparable to C57Bl/6.MRL-lpr/lpr and MRL-lpr/lpr mice, suggesting that reciprocal mutations in either fas or fas ligand increases susceptibility to bacterial superantigens (SAGs) . SEB-induced lethal shock can be reversed by treatment with Abs to V beta 8 TCR, MHC class II Ia+, IL-2, and TNF-alpha, by the immunosuppressant cyclosporin A, or by treatment with carbocyclic nucleoside analogues . These data indicate that SAG-induced septic shock is dependent on interactions with the TCR and MHC class II Ags, and they also suggest a critical role for a functional fas and/or fas ligand in resistance to SAG-induced septic shock. J Mol Biol, 1995 Nov 3, 253(4), 559 - 75 The crystal structure of the antibody N10-staphylococcal nuclease complex at 2.9 A resolution; Bossart-Whitaker P et al.; The three-dimensional structure of the antibody N10 Fab fragment complexed with staphylococcal nuclease (SNase) has been determined to 2.9 A resolution . Eighteen residues from six complementarity-determining regions (CDR) recognize an epitope of five distinct SNase segments with a total of 17 residues . The overall shape of the antibody-antigen interface is U-shaped rather than the more or less rectangular interface seen in other antibody-protein antigen interfaces . Despite the U-shaped interface, the amount of surface buried in the complex, 828 A2 for SNase and 793 A2 for N10, is typical of antibody-protein antigen complexes . Contributing to the shape of the interface is the shortest antibody heavy chain-CDR3 loop reported to date, which probably allows access of bulk solvent in the center of the "U" interface . Another unusual feature of the N10 antibody is the 15 residue antibody light chain-CDR1, a length seen in only three other reported antibodies . Antibody light chain-CDR1 displays a previously unobserved conformation in its distal portion . Finally, although some of the movement observed in the antibody-bound SNase may be due to crystal contacts, it is clear that some side-chain rearrangements are the result of antigen-antibody interaction. Eur Heart J, 1995 Nov, 16(11), 1686 - 91 Is the clinical spectrum and prognosis of native valve infective endocarditis in non-addicts changing? Tornos MP, Olona M, Permanyer-Miralda G, Almirante B, Evangelista A, Soler-Soler J. One hundred and ninety-four episodes of endocarditis on native valves in non-addict patients were diagnosed from 1975 to 1992 and were divided into groups A (78 patients, 1975-1983) and B (116 patients 1984-1992) . Both groups had the same gender distribution, similar valvular involvement and microbiological characteristics . In group B patients, median age was older (46 vs 54 years, P = 0.0002), the number of patients without previous heart disease was higher (46% vs 22%, P = 0.02) and the median time of symptoms before diagnosis was shorter (30 vs 50 days, P = 0.038) . Both groups had similar incidence of heart failure (32% vs 36%), surgical treatment (30% vs 33%) and embolic episodes (26% vs 34%) . Surgical mortality decreased from 43% to 18% (P = 0.03) . Overall mortality decreased non-significantly from 19% in group A to 12% in group B . Predictors of death in group A were heart failure (odds ratio 9.6, 95% confidence interval 3-36) and surgical treatment (odds ratio 5, 95% confidence interval 1.3-19) . Predictors of death in group B were age (odds ratio 4.98, 95% confidence interval 1.4-19), female sex (odds ratio 5.3, 95% confidence interval 1.3-20), staphylococcal infection (odds ratio 4.9, 95% confidence interval 1.1-22) and heart failure (odds ratio 5.2, 95% confidence interval 1.3-20) . Although in recent years infective endocarditis occurs in older patients and is more common in patients with previously unknown heart disease a substantial change in major clinical and prognostic variables is not apparent in our population . Overall in-hospital mortality has decreased from 19% to 12% mainly due to better surgical results. Hepatogastroenterology, 1995 Nov-Dec, 42(6), 816 - 20 Bacteremia associated with extracorporeal shockwave lithotripsy of gallbladder stones; Kullman E et al.; BACKGROUND/AIMS: The possible induction of bacteremia by extracorporeal shock wave lithotripsy (ESWL) of gallbladder stones was studied . MATERIALS AND METHODS: Seventy-six patients undergoing a total of 107 ESWL treatments were studied . RESULTS: Twenty-four (22%) of the 107 treatments were associated with bacteremia . Staphylococcus epidermidis was cultured during and/or after 23 (96%) of the treatments associated with bacteremia . The ESWL-induced tissue damage of the skin in the pass-way of the shock-waves was the most likely cause of bacteremia in these patients . There was no correlation between the occurrence of bacteremia and the age or body mass index of the patients . Neither was there any correlation of bacteremia related to the duration of the treatment, the number of shock waves, the energy delivered, the stone volume or the occurrence of calcified stones . No patient developed sepsis or endocarditis . Transient fever shortly after treatment was recorded in 5 patients (5%), one of whom had bacteremia . CONCLUSIONS: Routine antibiotic prophylaxis is not indicated in patients undergoing ESWL for gallbladder stones . The question whether such prophylaxis should be given to patients at special risk, for instance patients with artificial heart valves or known valvular heart disease, remains to be answered in larger controlled and randomized studies. Int J STD AIDS, 1995 Nov-Dec, 6(6), 447 - 9 Spectrum of opportunistic infections among AIDS patients in Tamil Nadu, India; Kumarasamy N et al.; A retrospective case note review of 100 AIDS patients attending a large Indian centre was performed . Of these 100 patients, 94% gave a history of heterosexual HIV transmission, 68% were male . The majority of females were aged 21 to 30 years . The most common mode of presentation was tuberculosis (61%), both pulmonary (46%) and extrapulmonary (15%) . Oral candidiasis extending on to the oesophagus was the second most predominant opportunistic infection . This study also highlights the difficulty in detecting AIDS cases in India owing to difficulties in taking a sexual history and lack of laboratory facilitiesPIP: In India, physicians examined and screened 100 AIDS patients aged 12-55 admitted to Government General Hospital in Madras for opportunistic infections as part of a study to document the characteristics of AIDS patients in Tamil Nadu State . 58% were 21-30 years old . The male/female ratio was 2:1 . 94% had acquired HIV via heterosexual intercourse . 81% of all patients had multiple sex partners and unprotected penetrative sex . Around 66% had more than one opportunistic infection . The most common opportunistic infection was tuberculosis (61%), especially pulmonary tuberculosis (46%), followed by oral candidiasis (41%), cryptosporidial diarrhea (16%), and fungal infection of the skin (16%) . The tuberculosis in most AIDS patients was reactivation of previously acquired tuberculosis . All tuberculosis patients responded well to standard treatment . The most common organism causing opportunistic infections was Staphylococcus pyogenes . Obstacles to acquiring more information about characteristics of these AIDS patients included the taboo of talking about sex and limited laboratory facilities . Clinicians should consider HIV in the differential diagnosis and management of all persons with tuberculosis . Zhonghua Yu Fang Yi Xue Za Zhi, 1995 Nov, 29(6), 344 - 7 {Observational and experimental studies on relationship between domestic pigs and epidemic hemorrhagic fever investigation}; Zhang Y et al.; Observational and experimental studies on the relationship between domestic pigs and epidemic hemorrhagic fever (EHF) were carried out during 1986 to 1994 with reverse passive hemagglutination (RPHA), reverse passive hemagglutination inhibition (RPHI), horse-radish peroxidase-staphylococcal protein A (HRP-SPA), indirect immunofluorescence assay (IFA) and reverse transcript polymerase chain reaction (RT-PCR) to understand the roles of domestic pigs played in the epidemic areas . Results showed domestic pigs could infect epidemic hemorrhagic fever virus (EHFV) naturally or experimentally, and EHFV could disseminate to many organs of the animals and caused transient pathologic changes in them . EHFV could duplicate within the animal bodies and be excreted out of the bodies with their excreta . Proportion of inapparent infection of EHF in people closely exposed to domestic pigs was significantly higher than in those without it . It indicated domestic pigs maybe played an important role in EHF as a reservoir host. Immunopharmacology, 1995 Nov, 31(1), 109 - 16 Thalidomide increases the synthesis of IL-2 in cultures of human mononuclear cells stimulated with Concanavalin-A, Staphylococcal enterotoxin A, and purified protein derivative; Shannon EJ et al.; Thalidomide significantly increases the quantity of extracellular IL-2 in cultures of human mononuclear cells stimulated with mitogens or antigen . Cells from 7 donors exposed for 2 h to 4.0 micrograms/ml of thalidomide and stimulated for 16-18 h with 20 micrograms/ml of Concanavalin-A (Con-A) averaged producing 187 +/- 49% more IL-2 than cells stimulated with Con-A alone . In similar experimental procedures and comparisons the pg/ml of IL-2 secreted by thalidomide-treated cells from five donors stimulated with 50 ng/ml of Staphylococcal enterotoxin A (SEA) increased by 159 +/- 32%, and the pg/ml of IL-2 secreted by thalidomide-treated cells from 2 donors stimulated with 5.0 micrograms/ml of purified protein derivative of Mycobacterium tuberculosis increased by 120 +/- 4% . Thalidomide also significantly increases the quantity of intracellular IL-2 in cells stimulated with mitogens . Cells exposed to thalidomide and stimulated with Con-A had an increase in intracellular IL-2 of 130% after 8 h and 157% after 12 h in culture; cells stimulated with SEA had an increase in intracellular IL-2 of 120% after 8 h and 182% after 12 h in culture . Thalidomide did not alter the percent of lymphocytes expressing the alpha-chain of IL-2 receptor, nor did it significantly increase incorporation of {3H}thymidine by cells. Bioorg Med Chem, 1995 Nov, 3(11), 1519 - 25 Design, synthesis, and biological evaluation of isocyanurate-based antifungal and macrolide antibiotic conjugates: iron transport-mediated drug delivery; Ghosh M et al.; The syntheses and preliminary biological evaluation of conjugates of a synthetic isocyanurate-based trihydroxamate siderophore with two antifungal agents, 5-FU (conjugate 9) and norneoenactin (conjugate 12), and a macrolide antibiotic, erythromycylamine (conjugate 18), are described . A 19F NMR study was used to determine the hydrolytic stability of conjugate 9 under assay conditions . Preliminary biological studies with ferric complexes of conjugates 9 and 12 indicated that these antifungal agents are recognized by Candida and perhaps are actively transported into the cell by the siderophore-transport mechanisms . While conjugate 18 did not show any significant antibacterial activity, presumably due to size restriction, the 5-FU conjugate 9 appeared to be moderately active against a variety of Gram-positive strains, and was more active than the 5-FC control against some strains of Staphylococcus. J Clin Immunol, 1995 Nov, 15(6 Suppl), 37S - 41S Modification of immediate hypersensitivity responses by staphylococcal enterotoxin B; Gelfand EW et al.; The staphylococcal enterotoxins have been termed superantigens based on their ability to stimulate polyclonal proliferative responses of murine and human T lymphocytes expressing particular T-cell receptor V beta gene products . Certain of these toxins have been shown both to activate and to induce anergy in reactive T cells . Staphylococcal enterotoxin B is known to interact with murine T cells bearing V beta 3, -7, -8.1, -8.2, -8.3, and -17 . In BALB/c mice V beta 3+ and V beta 17+ T cells are deleted; V beta 7+ T cells are low in frequency . BALB/c mice sensitized to ovalbumin via the skin and airways develop immediate hypersensitivity including IgE/IgG1 antiovalbumin antibodies, immediate cutaneous reactivity to ovalbumin and, increased airway responsiveness . In both in vitro and in vivo studies, the development of these responses has been associated with the V beta 8+ subset of T cells and controlled by V beta 2 + T cells . In view of the central role of V beta 8+ T cells in these responses, we tested the effects of staphylococcal enterotoxin B on the development of immediate hypersensitivity in this system . Intradermal injection of staphylococcal enterotoxin B prevented the development of these responses in the absence of a major deletion of V beta 8+ T cells . The data suggest that the administration of staphylococcal enterotoxin B prevented the antigen-induced expansion of V beta 8+ T cells resulting in a state of responsiveness or anergy, thus preventing the manifestations of immediate hypersensitivity . Bacterial toxins may provide a novel approach to intervention in allergic or autoimmune diseases. J Clin Immunol, 1995 Nov, 15(6 Suppl), 26S - 36S B-cell superantigens: definition and potential impact on the immune response; Levinson AI et al.; Superantigens have been extremely helpful tools in exploring fundamental questions in immunobiology including mechanisms of cell activation, tolerance, and autoimmunity . Until recently, attention has been focused exclusive on T-cell superantigens . However, new data suggest that there are superantigens that directly activate B cells . By definition, these agents (1) stimulate a high frequency of B cells, (2) target B cells that have restricted usage of VH or VL family genes, and (3) bind to immunoglobulins outside the sites that bind conventional antigens . A candidate B-cell superantigen that has received considerable attention in this laboratory is staphylococcal protein A . This agent is best known to the immunologist because of its ability to bind to the Fc fragment of IgG . This binding has been localized to two alpha-helical structures on each of four or five homologous regions that comprise the extracellular domain of protein A . However, it is now clear that protein A contains a second site that binds to determinants on the Fab regions of certain immunoglobulins independently of their heavy-chain isotype . In man this so-called alternative site appears to bind only to immunoglobulins that utilize heavy-chain genes of the VH3 subfamily . In the mouse this type of binding is restricted to immunoglobulins using heavy chains belonging to the S107 and J606 VH families . In this review, we examine the growing list of microbial products that dominate B-cell superantigenic properties . Using staphylococcal protein A as a model for a B-cell superantigen, we consider the potential impact of this novel class of antigens on the immune response . We focus on the ability of B-cell superantigens to influence the expression of the B-cell repertoire . In addition, we consider the hypothesis that the interaction of a B-cell superantigen with its reactive serum immunoglobulins activates the classical complement cascade and thus represents a powerful stimulant of tissue inflammation. Int Immunol, 1995 Nov, 7(11), 1763 - 9 Resident CD4+ alpha beta T cells of the murine female genital tract: a phenotypically distinct T cell lineage that rapidly proliferates in response to systemic T cell activation stimuli; Ibraghimov AR et al.; A population of CD4+ cells has been identified in the murine female genital tract (FGT) . Phenotypic studies of FGT CD4+ cells demonstrate that they express CD3 and that the majority of these cells are alpha beta TCR+Thy-1+ . Most of the Thy-1+CD4+alpha beta TCR+ cells resemble memory T cells based on their expression of CD44, L-selectin and CD45RB antigens . The vast majority of Thy-1+CD4+alpha beta TCR+ FGT cells are CD5+ and all of them are B220- . Systemic stimuli including infection with Trypanosoma brucei brucei, injection with anti-CD3 epsilon, or bacterial superantigens staphylococcal enterotoxin A or B cause a rapid accumulation of CD4+ cells in the FGT exceeding that observed for CD4+ cells in spleen and lymph nodes (LN) . Expansion of the FGT CD4+ cells, which are phenotypically distinct from the splenic and LN CD4+ T cells, is due to local proliferation rather than an influx of cells from the circulation . The CD4+ population in the FGT of adult nu/nu mice is dramatically reduced, indicating its thymic dependency . In lpr/lpr mice, FGT CD4 cells do not display changes characteristic of splenic or LN CD4 cells in the same animals . These findings demonstrate that the CD4+ cells of the murine FGT are thymic dependent, but that they constitute a T cell lineage that phenotypically and, probably functionally, is distinct from other peripheral CD4+ T cell populations. Int Immunol, 1995 Nov, 7(11), 1721 - 7 Characterization of an alternative superantigen binding site expressed on a renal fibroblast cell line; Rogers TJ et al.; It is well established that the bacterial superantigens bind to both TCR beta-chain and, with moderate affinity, to MHC class II molecules . Class II-bearing cells bind the superantigen and present the superantigen to T cells expressing certain TCR beta-chain variable region alleles . We have observed that the superantigen staphylococcal enterotoxin (SE) B binds to COS-1, an African green monkey kidney fibroblast-like cell line . This cell line fails to express class II and the binding of SEB is saturable . Scatchard analysis of radiolabeled ligand binding data reveals a binding affinity for COS of 51 nM, while binding to DR1-transfected L cells is measured at 150 nM . Further analysis shows that SEB bound to COS cells, unlike SEB bound to DR1-bearing L cells, is not recognized by T cells in the presence or absence of accessory cytokines or anti-CD28 . Studies carried out with chemical cross-linking agents show that radiolabeled SEB associates with a membrane protein of approximately 85 kDa . These studies suggest that the alternative superantigen binding molecule (p85) binds SEB with an affinity roughly equivalent to SEB binding by class II molecules, but at a site which does not permit recognition by the TCR. Cutis, 1995 Nov, 56(5), 276 - 8 Generalized pustular eruption associated with converting enzyme inhibitor therapy; Carroll J et al.; A 67-year-old man presented with a high fever and a generalized rash . His extended hospital stay was characterized by fever with repeated staphylococcal bacteremia and the appearance of axillary lymphadenopathy and splenomegaly . Skin lesions became hyperpigmented, dry, and atrophic with areas of exfoliation and uclers . Examination of skin and lymph node biopsy specimens showed findings consistent with mycosis fungoides . The patient unexpectedly recovered on discontinuation of captopril . A positive macrophage inhibiting factor response for both captopril and enalapril indicated that the non-sulfhydryl moiety was the antigenic stimulant for the lesion resembling mycosis fungoides. J Gastroenterol, 1995 Nov, 30 Suppl 8, 73 - 5 CD4+ intestinal mucosal lymphocytes in the pathogenesis of Crohn's disease; Watanabe M et al.; To analyze the nature of intestinal mucosal lymphocytes in Crohn's disease, we established T cell lines of patients' intraepithelial lymphocytes . T cell lines from the affected terminal ileum of the patients showed an increased proportion of CD4+V beta 5.2/5.3+ T cells . These cells were increased in number after stimulation with staphylococcal enterotoxins C1 and D, showed an increase in cytolytic activity, and produced a large amount of interferon-gamma . To clarify the role of CD4+ mucosal lymphocytes in the intestinal inflammation, we then developed a novel colitis model by immunizing a rat with trinitrobenzenesulfonic acid (TNB) emulsion with adjuvant . Deep ulceration and granuloma formation in this colitis model resembled the histopathological findings of human Crohn's disease . Immunohistochemical and flow cytometric analysis demonstrated that the number of CD45RC(high)CD4+ mucosal lymphocytes was increased . Interestingly, the administration of anti-CD4 Abs prevented severe inflammation in the model . After treatment with anti-CD4 Abs, the anti-TNB Ab titer, the number of CD45RC(high)CD4+ cells, and interferon-gamma mRNA expression were significantly decreased in the mucosa of the model . These results suggest that some subsets of CD4+ mucosal lymphocytes play an important role in the triggering and progression of inflammation in Crohn's disease. Appl Environ Microbiol, 1995 Nov, 61(11), 3894 - 903 Isolation and characterization of genetically engineered gallidermin and epidermin analogs; Ottenwalder B et al.; Gallidermin (Gdm) and epidermin (Epi) are highly homologous tetracyclic polypeptide antibiotics that are ribosomally synthesized by a Staphylococcus gallinarum strain and a Staphylococcus epidermidis strain, respectively . These antibiotics are secreted into media and are distinguished by the presence of the unusual amino acids lanthionine, 3-methyllanthioni