J Pediatr Surg, 2000 Jan, 35(1), 120 - 3 Patch incorporation in diaphragmatic hernia; Kimber CP et al.; BACKGROUND/PURPOSE: Biomaterial insertion often is required for closure of congenital diaphragmatic hernia (CDH) . The optimal biomaterial remains uncertain . This study was designed to compare a commonly used patch (polytetrafluoroethylene) with a recently available fabric, fluorinated polyester . The aim of this study was to determine the clinical performance, histological tissue-polymer interaction, bacterial adhesion, and shrinkage rates of biomaterial inserted endoscopically into a CDH lamb model . METHODS: Polytetrafluoro-ethylene (PTFE) and fluorinated polyester (FP) were randomised for laparoscopic patch insertion into 12 lambs . All lambs (age <4 weeks) underwent 3-port laparoscopy, surgical creation of diaphragmatic hernia, and sutured patch placement . Two PTFE and 2 FP lambs were killed at 1-, 3-, and 6-month intervals postoperatively . Postmortem examination histopathology, electron microscopy, and specific bacterial broth immersion (Escherichia coil, Staphylococcus aurens, and epidermidis) were performed . RESULTS: All 12 lambs completed the study with intact patches that were fully peritonised . One abdominal adhesion was noted in a FP lamb at 6 months . FP was comparatively easier to insert, manipulate, and suture endoscopically . Histopathology findings showed that PTFE patches created a strong peripheral foreign body reaction with dystrophic calcification, whereas FP was well incorporated with intrapatch fibroblastic activity and neovascularsation . No significant difference in resistance to bacterial adhesion of relevant organisms was noted between the materials . Graft shrinkage for FP was 7% in one direction only, evident by 3 months . CONCLUSIONS: Fluorinated polyester has advantages in this laparoscopic lamb model . It shows rapid and sustained incorporation with intrapatch neovascularisation when compared with polytetrafluoro-ethylene's significant foreign body reaction . It was preferred for its endoscopic handling and suturing properties . The laparoscopic techniques used may contribute to the general lack of adhesions, and insufficient data are available to comment on the comparative effect of the materials on adhesion formation . No difference was demonstrated in resistance to bacterial adherence in the harvested materials. Vojnosanit Pregl, 1999 Sep-Oct, 56(5), 541 - 5 {Staphylococcal pneumonia associated with pneumatoceles, bilateral pyopneumothorax and sepsis}; Stanic V et al.; A case of soldier with community acquired staphylococcal pneumonia and multiple pneumatoceles as the rare complication in adults is presented . Their recognition provides appropriate treatment . In the patient were developed bilateral pneumothorax, pleural empyema and sepsis . Surgical treatment was performed by thoracic drainage . Recovery of pulmonary function was poor, as distinguished from children. J Immunol, 2000 Feb 1, 164(3), 1175 - 84 IL-2 unresponsiveness in anergic CD4+ T cells is due to defective signaling through the common gamma-chain of the IL-2 receptor; Grundstrom S et al.; Repeated administration of the superantigen staphylococcal enterotoxin A to mice transduces a state of anergy in the CD4+ T cell compartment, characterized by inhibition of IL-2 production and clonal expansion in vivo . In contrast to what has been reported on anergic T cell clones in vitro, culture of in vivo anergized CD4+ T cells in the presence of exogenous IL-2 did not overcome the block in responsiveness . In this study, we demonstrate that CD4+ T cells from mice anergized with staphylococcal enterotoxin A also exhibit a reduced proliferative capacity in response to IL-7 and IL-15, cytokines that share a common gamma-chain with the IL-2R . Flow-cytometric analysis revealed only modest changes in the expression of the different IL-2R chains . In a number of experiments, our results also provide evidence that excludes a major role of the IL-2R alpha-chain in this system . According to these results, the inability of anergic cells to respond to IL-2 is not mainly due to a down-regulation of the high affinity IL-2R, but to a perturbation in intracellular signaling . Our study confirmed that the activation and tyrosine phosphorylation of Janus-associated kinase 3 and STAT5 were considerably weaker after anergy induction . Moreover, anergic CD4+ T cells showed significantly reduced DNA-binding ability to STAT5-specific elements . Taken together, we suggest that the observed IL-2 unresponsiveness in anergic CD4+ T cells could be due to a defect in signaling through the common gamma-chain of the IL-2R. J Biol Chem, 2000 Jan 21, 275(3), 1665 - 72 The spectral and thermodynamic properties of staphylococcal enterotoxin A, E, and variants suggest that structural modifications are important to control their function; Cavallin A et al.; The superantigens staphylococcal enterotoxin A and E (SEA and SEE) can activate a large number of T-cells . SEA and SEE have approximately 80% sequence identity but show some differences in their biological function . Here, the two superantigens and analogues were characterized biophysically . SEE was shown to have a substantially higher thermal stability than SEA . Both SEA and SEE were thermally stabilized by 0.1 mM Zn(2+) compared with Zn(2+)-reduced conditions achieved using 1 mM EDTA or specific replacements that affect Zn(2+) coordination . The higher stability of SEE was only partly caused by the T-cell receptor (TCR) binding regions, whereas regions in the vicinity of the major histocompatibility complex class II binding sites affected the stability to a greater extent . SEE exhibited a biphasic denaturation between pH 5.0-6.5, influenced by residues in the TCR binding regions . Interestingly, enzyme-linked immunosorbent assay, isoelectric focusing, and circular dichroism analysis indicated that conformational changes had occurred in the SEA/E chimerical constructs relative to SEA and SEE . Thus, it is proposed that the Zn(2+) binding site is very important for the stability and potency of SEA and SEE, whereas residues in the TCR binding site have a substantial influence on the molecular conformation to control specificity and function. J Neuroimmunol, 2000 Jan 24, 102(2), 131 - 6 Superantigen presenting capacity of human astrocytes; Hassan-Zahraee M et al.; We found that human fetal astrocytes (HFA) are able to support superantigen (SAG) staphylococcal enterotoxin B (SEB) and toxic shock syndrome toxin-1 (TSST-1)-induced activation of immediately ex vivo allogenic human CD4 T cells . Using radiolabelled toxins, we demonstrate that both SEB and TSST-1 bind with high affinity to MHC class II antigen expressing astrocytes; binding is displaceable with excess cold toxin . Competition experiments further indicate that TSST-1 and SEB at least partially compete with each other for binding to astrocytes suggesting they bind to the same HLA-DR region on these cells . Our study supports the hypothesis that SAG would be capable of stimulating immune responses within the human CNS and contribute to persistence or recurrence of inflammatory responses within this compartment. J Nephrol, 1999 Sep-Oct, 12(5), 318 - 21 Outcome of peritonitis treated with intraperitoneal (i.p.) weekly vancomycin and i.p . daily netilmicin; Chadwick DH et al.; The treatment of peritoneal dialysis related peritonitis has been streamlined by the recommendations of an ad hoc committee, which has advocated avoidance of vancomycin as an empirical therapy because of the prevailing problems of vancomycin resistant enterococci (VRE) . This report relates to the continued use of vancomycin combined with netilmicin in our programme as empiric therapy for CAPD peritonitis and reports on favorable results of over 80% cure rate using this regimen . The need for change to vancomycin avoiding regimes has to be related to the knowledge of local prevalence of VRE and staphylococcal sensitivity patterns. Toxicol Sci, 1999 Dec, 52(2), 189 - 98 Modulation of the activity of human monocyte-derived dendritic cells by chemical haptens, a metal allergen, and a staphylococcal superantigen; Coutant KD et al.; For the development of mechanistic assays in immunotoxicology, the phenotype, cytokine production, and stimulatory function of dendritic cells (DCs) were assessed after incubation with the chemical haptens aminophenol, chlorpromazine hydrochloride, dinitrochlorobenzene (DNCB), and with the DNCB-corresponding tolerogen DCNB, the metal allergen nickel sulfate, the irritants sodium dodecyl sulfate and benzoic acid, as well as with staphylococcal enterotoxin B (SEB) and lipopolysaccharide (LPS) . DCs were differentiated from human monocytes by in vitro exposure to GM-CSF and interleukin-4 (IL-4) for 7 days . Flow cytometric data revealed that only representative haptens increased the surface expression of HLA-DR, CD86, CD40, and of CD54 on DCs when compared to irritants or to the tolerogen . This event was associated with an increased ability of DCs to stimulate T cell proliferation . Moreover, after incubation with the haptens, but not with the irritants or the tolerogen, a higher production of TNF-alpha by DCs was observed . Under our experimental conditions, no release of IL-1beta, IL-10, or IL-12 was detected . Compared to the activation elicited by haptens, SEB strongly up-regulated HLA-DR and costimulatory molecule expression . In agreement with this effect, there was a marked release of TNF-alpha and a slight production of IL-12 . IL-1beta and IL-10 were not detected in the culture medium . Finally, SEB-pulsed DCs showed a strong T-cell-stimulating activity . These data underline the activating potential of haptens versus irritants or a tolerogen on DC functions . The different levels of DC activation by haptens and SEB suggested that distinct cellular events were involved. Dermatol Clin, 2000 Jan, 18(1), 73 - 8, viii New and emerging therapies in pediatric dermatology; Raimer SS; Many of the dermatologic conditions for which children seek medical attention are caused by infectious organisms . Several medications have recently become available or are on the horizon for the treatment of pediatric skin infections and infestations . Treatment of tinea capitis with fluconazole, itraconazole, and terbinafine, antibiotic therapy for staphylococcal skin infections, cidofovir for the treatment of verrucae vulgaris and molluscum contagiosum and ivermectin for scabies and head lice are discussed. Sante, 1999 Jul-Aug, 9(4), 209 - 13 {Pleuropulmonary staphylococcal infection in infants, in a hospital environment in Ouagadougou (Burkina Faso)}; Sawadogo A et al.; We observed 36 cases of pleuropulmonary staphylococcal infection (PPS) in infants aged 0 to 30 months, during a prospective study carried out between April 1st 1995 and March 31 1996 at the Pediatrics Department of Ouagadougou University Hospital . PPS accounted for 0.5% of all hospital admissions and 11.6% of all acute basal respiratory infections in children aged less than 30 months . Slightly more boys than girls were affected, with a sex ratio of 1.2 . We identified the classic triad of symptoms: cough-fever-polypnea, associated with abdominal ballooning and a change in general condition . On X rays, the typical images showing parenchymatous bubbles were the second most frequent observation (27.8%) after parenchymatous opacities (69.5%) . The most frequently used antibiotics were oxacillin (Bristopen), gentamycin (Gentallin) and cefuroxime-axetil (Zinnat) . The prognosis of PPS is poor, with a high mortality rate (27.8%) and a risk of pleural recurrence . Being very young, late hospitalization, malnutrition and leukopenia were identified as factors indicating a poor prognosis . Recygling of health care personnel for the management of acute respiratory infections, a decrease in malnutrition and an improvement in vaccination cover are essential if the mortality and morbidity of acute respiratory infections, and PPS in particular, are to be reduced. J Immunol, 2000 Jan 15, 164(2), 754 - 61 The multidrug resistance protein 1: a functionally important activation marker for murine Th1 cells; Prechtl S et al.; Previously, we described the expression of an energy-dependent pump in resting murine Th2 (but not resting Th1) cells which extruded the fluorescent dye Fluo-3 . After stimulation with Ag and APCs, Th1 cells also expressed this pump . Furthermore, expression of the murine multidrug resistance protein 1 (mrp1) correlated with the presence of the pump . In this study, we report that Fluo-3 is indeed transported by murine mrp1 or its human ortholog MRP1, as revealed by transfection of HEK 293 cells with mrp1 or MRP1 cDNA . Like antigenic activation, IL-2 dose-dependently enhanced the Fluo-3-extruding activity in murine Th1 cells . Although TNF-alpha and IL-12 by themselves only weakly enhanced Fluo-3 extrusion, each of them did so in strong synergism with IL-2 . An Ab directed against mrp1 was used to quantify the expression of mrp1 protein in T cells at the single-cell level . Like the Fluo-3 pump, mrp1 protein expression was enhanced by IL-2 . Immunohistochemical studies using confocal laser microscopy indicated that mrp1 is localized mainly at the plasma membrane . In addition, protein expression of mrp1 was induced in Vbeta8+CD4+ T cells 12 h after in vivo application of Staphylococcal enterotoxin B . Finally, mrp1 was functionally relevant during the activation process of Th1 cells, because T cell activation could be suppressed by exposure of cells to the mrp1 inhibitor MK571 . Thus, we present mrp1 as a novel, functionally important activation marker for Th1 cells and short-term in vivo activated CD4+ T cells, whereas its expression seems to be constitutive in Th2 cells. Clin Infect Dis, 2000 Jan, 30(1), 146 - 51 Use of linezolid, an oxazolidinone, in the treatment of multidrug-resistant gram-positive bacterial infections; Chien JW et al.; We report our experience with linezolid in an investigation of its use against resistant gram-positive bacterial infections . Fifteen patients who had renal failure (n=6), recent liver transplantation (n=5) or surgery (n=6), cancer (n=3), endocarditis (n=2), or human immunodeficiency virus infection (n=1), along with infections due to vancomycin-resistant enterococcus (VRE), and 2 patients with infections due to methicillin-resistant Staphylococcus species who had adverse reactions to vancomycin were treated with linezolid (600 mg every 12 h for 5-42 days (mean+/-SD, 20.5+/-3.5 days) . Abscess drainage or prosthetic device removal was undertaken . Microbiological cure occurred in all 10 patients who completed therapy, and all 7 patients alive at follow-up were free of infection . No deaths were attributable to the index infection . Adverse events associated with linezolid use were mild leukopenia in 1 patient and nausea in another . It appears that administration of linezolid, in conjunction with surgical intervention or device removal, is an effective treatment option for serious resistant gram-positive bacterial infections. Postgrad Med J, 1999 Sep, 75(887), 540 - 3 Aetiology and outcome in 53 cases of native valve staphylococcal endocarditis; Hricak V et al.; Within the last 30 years the profile of infective endocarditis has altered considerably with regard to microbiological causation, clinical features, and natural history . A contributory factor has undoubtedly been the development of potent antibiotics and their sometimes indiscriminate use . The increase in intravenous drug abuse in urban centres, the use of immunosuppressive agents, and the use of prosthetic heart valves have also all contributed . Although cardiac surgery in the uninfected heart provides a perfect environment for infective endocarditis, the improved design of prosthetic valves and the enhanced long-term survival and decreased immediate operative risk, means that surgery is viewed as the best option in many cases . In a series of 53 cases of staphylococcal endocarditis from a national endocarditis survey, those risk factors which influenced outcome were analysed . Thirty out of 53 patients had predisposing heart disease . Mortality was 39.6% . Statistical analysis revealed that attributable mortality was significantly associated with skin infection, systemic embolisation, and inappropriate therapy . Interestingly, surgical treatment was associated with better outcome. Mt Sinai J Med, 1999 Oct-Nov, 66(5-6), 320 - 6 Hickman-Broviac catheter-related infections in children with malignancies; Stamou SC et al.; Infectious complications are frequently encountered following Hickman-Broviac (H-B) catheter insertion . The medical records of 164 children with malignancies who underwent H-B catheter insertion from March 1, 1988 to December 31, 1997 were reviewed retrospectively . During a 35,697 catheter-day period, 77 catheter-related infections occurred, including 50 catheter-insertion-site infections and 27 bloodstream infections . The risk for the development of catheter-related infections was 2.15 per 1000 catheter-days (1.4 and 0.75 per 1000 catheter-days for catheter-insertion-site and bloodstream infections, respectively) . In 17 (63%) of 27 episodes of bloodstream infections, antimicrobial treatment controlled the infection without catheter removal . A previous catheter-insertion-site infection caused by Staphylococcus epidermidis (p=0.01), the occurrence of mechanical catheter complications (p=0.007), and a normal coagulation status of the host (p=0.03) were significantly associated with the development of catheter-related bloodstream infections . H-B catheters remain important in pediatric oncology . Due to the significant morbidity associated with the development of catheter-related bloodstream infections, risk factors found to increase the incidence rate of such infections must be identified and properly managed. Proc Natl Acad Sci U S A, 2000 Jan 4, 97(1), 133 - 8 Staphylococcal protein A: unfolding pathways, unfolded states, and differences between the B and E domains; Alonso DO et al.; We present multiple native and denaturation simulations of the B and E domains of the three-helix bundle protein A, totaling 60 ns . The C-terminal helix (H3) consistently denatures later than either of the other two helices and contains residual helical structure in the denatured state . These results are consistent with experiments suggesting that the isolated H3 fragment is more stable than H1 and H2 and that H3 forms early in folding . Interestingly, the denatured state of the B domain is much more compact than that of the E domain . This sequence-dependent effect on the dimensions of the denatured state and the lack of correlation with structure suggest that the radius of gyration can be a misleading reaction coordinate for unfolding/folding . Various unfolding and refolding events are observed in the denaturation simulations . In some cases, the transitions are facilitated through interactions with other portions of the protein-contact-assisted helix formation . In the native simulations, the E domain is very stable: after 6 ns, the C(alpha) root-mean-square deviation from the starting structure is less than 1.4 A . In contrast, the native state of the B domain deviates more and its inter-helical angles fluctuate . In apparent contrast, we note that the B domain is thermodynamically more stable than the E domain . The simulations suggest that the increased stability of the B domain may be due to heightened mobility, and therefore entropy, in the native state and decreased mobility/entropy in the more compact denatured state. Ryumachi, 1999 Oct, 39(5), 757 - 62 {A case of microscopic polyangiitis with severe cardiac and respiratory muscle involvement}; Sendoh W et al.; A 66-year-old female was admitted to our hospital in January, 1998, complaining of low grade fever and muscle weakness of her legs . Physical examination revealed muscle weakness of her neck (4/5) and proximal skeletal muscles of her bilateral legs (3/5-4/5) . She showed proteinuria and microhematuria . Her serum levels of ureanitrogen, creatinine, aspartate aminotransferase, alanine aminotransferase, creatinekinase, aldolase and myoglobin were all within the normal ranges . Antinuclear antibodies were negative, but her serum levels of pANCA (743 EU) and C reactive protein (18.0 mg/dl) were elevated . Neuroconduction velocity of her left common peroneal nerve was decreased to 40.8 m/sec and electric myograph showed neurogenic changes . Magnetic resonance images (MRI) of her bilateral thigh depicted high signal intensity in quadriceps by T 2 weighed images, but the signals were not enhanced by gadolinium injection . Muscle and renal biopsies revealed necrotizing vasculitis of the small arteries . Crescentic glomerulonephritis was also observed by renal biopsy . These findings supported the diagnosis of microscopic PN . On 16 th admission day, she developed acute cardiac and respiratory failures due to cardiac and respiratory muscle involvements with PN, and was assisted by mechanical ventilation . She was treated with methylprednisolone pulse therapy (500 mg/day, three consecutive days) on 18 th admission day, followed by 40 mg of oral prednisolone daily . However, her symptoms deteriorated, and herserum creatinine levels increased to 2.4 mg/dl . On 24 th admission day, intravenous cyclophosphamide pulse therapy (500 mg/day) was instituted . Her cardiac wall motion on echocardiography and serum creatinine levels gradually improved, but her skeletal and respiratory muscle weakness did not improve . On 38 th admission day, she was complicated with respiratory infection by methicillin resistant Staphylococcus aures . On 62 th admission day, she died of endotoxic shock . This is the first report describing respiratory muscle involvement with PN, and the second report describing MRI findings of muscle involvement by PN . Therefore, our case provides important clinical information for the diagnosis and treatment of the disease. Proc Natl Acad Sci U S A, 1999 Dec 21, 96(26), 14848 - 53 Pressure-induced protein-folding/unfolding kinetics; Hillson N et al.; We use an off-lattice minimalist model to describe the effects of pressure in slowing down the folding/unfolding kinetics of proteins when subjected to increasingly larger pressures . The potential energy function used to describe the interactions between beads in the model includes the effects of pressure on the pairwise interaction of hydrophobic groups in water . We show that pressure affects the participation of contacts in the transition state . More significantly, pressure exponentially decreases the chain reconfigurational diffusion coefficient . These results are consistent with experimental results on the kinetics of pressure-denaturation of staphylococcal nuclease. Cell Immunol, 1999 Nov 1, 197(2), 129 - 35 Cross-linking staphylococcal enterotoxin A bound to major histocompatibility complex class I is required for TNF-alpha secretion; Wright AD et al.; The mechanism of how superantigens function to activate cells has been linked to their ability to bind and cross-link the major histocompatibility complex class II (MHCII) molecule . Cells that lack the MHCII molecule also respond to superantigens, however, with much less efficiency . Therefore, the purpose of this study was to confirm that staphylococcal enterotoxin A (SEA) could bind the MHCI molecule and to test the hypothesis that cross-linking SEA bound to MHCII-deficient macrophages would induce a more robust cytokine response than without cross-linking . We used a capture enzyme-linked immunosorbent assay and an immunprecipitation assay to directly demonstrate that MHCI molecules bind SEA . Directly cross-linking MHCI using monoclonal antibodies or cross-linking bound SEA with an anti-SEA antibody or biotinylated SEA with avidin increased TNF-alpha and IL-6 secretion by MHCII(-/-) macrophages . The induction of a vigorous macrophage cytokine response by SEA/anti-SEA cross-linking of MHCI offers a mechanism to explain how MHCI could play an important role in superantigen-mediated pathogenesis . Med Clin (Barc), 1999 Oct 23, 113(13), 488 - 9 {Oral antibiotic therapy in the adult bacterial osteomyelitis: results after two years of follow-up}; Javaloyas de Morlius M et al.; BACKGROUND: Oral antibiotic therapy achieves clinical and bacteriological cure of the adult bacterial osteomyelitis associated or not to orthopedic implant . PATIENTS AND METHODS: We carried out a prospective study, with follow-up at 24 months, of patients with adult bacterial osteomyelitis, that were initially treated with parenteral antibiotic therapy for a week, followed with oral antibiotic therapy during 2 to 6 months, depending on the absence or presence or orthopedic implant, respectively . RESULTS: 37 patients presented 44 episodes of adult bacterial osteomyelitis, 34 of them in presence of orthopedic implant . The most frequent causative microorganism was S . aureus (44%), followed by coagulase-negative Staphylococcus (29%) . The oral antibiotic therapy most frequently employed was the combination cotrimoxazole-rifampin, followed by ciprofloxacin-rifampin . The clinical and bacteriologic cure were 85%, 82% in the group with orthopedic implant . 82% required surgical intervention . CONCLUSIONS: The oral antibiotic therapy, preceded by a short parenteral therapy, can get high rates of clinical and bacteriological cure in the adult's bacterial osteomyelitis. J Thromb Thrombolysis, 1994, 1(1), 49 - 54 Heparin Binding Sites Are Located in a 40-kD gamma-Chain and a 36-kD beta-Chain Fragment Isolated from Human Fibrinogen; Mohri H et al.; Objective: We have previously shown that (125)I-fibrinogen binds to heparin sepharose CL-6B . To identify the localization of the heparin binding domain in human fibrinogen, reduced and alkylated fibrinogen was digested by limited-Staphylococcus aureus V8 protease . Methods/Results: Two fragments have now been isolated and purified to apparent homogeneity by heparin-affinity chromatography . These fragments, denoted the 40-kD and 36-kD fragments, contain NH(2)-terminal sequences of Ala-Ser-Ile-Leu-Thr-Hb;-Asp and Thr-Val-Asn-Ser-Asn-Ile-Pro, respectively . These fragments established the positions of these peptides within the gamma chain of fibrinogen as beginning with the residue tentatively designated 124 and within the beta chain as beginning with the residue designated 186 . Binding of (125)I-fibrinogen to heparin-sepharose CL-6B was completely inhibited by a mixture of these fragments, with an IC(50) of 3.2 microM . The synthetic peptide of the gamma chain carboxy-terminal 15 residues (GQQHHLGGAKQAGDV;G15) partially inhibited fibrinogen binding . The mixture of these fragments partially inhibited the ADP-induced aggregation of platelets . Conclusions: These data indicate that the domains for heparin binding may be present on both the gamma chain and the beta chain of fibrinogen, and that the domain on the gamma chain may be close to the binding domain on the carboxy terminus of the fibrinogen gamma chain to glycoprotein IIb-IIIa. Infect Immun, 2000 Jan, 68(1), 120 - 4 Induction of tumor necrosis factor alpha and interleukin-8 gene expression in bronchial epithelial cells by toxic shock syndrome toxin 1; Aubert V et al.; Major histocompatibility complex (MHC) class II engagement by toxic shock syndrome toxin 1 (TSST-1) transduces signals leading to proinflammatory cytokine gene expression (tumor necrosis factor alpha {TNF-alpha}) in human monocytes . To study the proinflammatory role of MHC class II molecules expressed by bronchial epithelial cells (BEC), primary human BEC were isolated from surgical bronchial samples, expanded in vitro, and cultured in the presence or absence of gamma interferon (IFN-gamma) for 48 h . (125)I-TSST-1 binding to BEC pretreated with IFN-gamma was inhibited up to 97% by anti-MHC class II monoclonal antibody 3B12, indicating that in BEC also MHC class II molecules were targets for the staphylococcal exotoxin . As analyzed by a quantitative reverse transcriptase PCR, a 1-h stimulation of BEC with TSST-1 resulted in a vigorous expression of TNF-alpha and interleukin-8 (IL-8) genes . TNF-alpha and IL-8 expression was optimal in BEC pretreated with 50 IU of IFN-gamma/ml, whereas TSST-1 stimulation of BEC pretreated with 200 IU of IFN-gamma/ml failed to enhance either TNF-alpha or IL-8 transcripts . In a time course study, peak expression of TNF-alpha and IL-8 mRNA was reached 6 h after TSST-1 stimulation . These results demonstrate that bacterial superantigen TSST-1 binds to MHC molecules on BEC and induces TNF-alpha and IL-8 gene expression upon engagement of MHC class II molecules on BEC, thus contributing to the perpetuation of bronchial mucosa inflammation via chemokine or cytokine gene expression. Biochemistry, 1999 Dec 14, 38(50), 16514 - 28 Effect of staphylococcal delta-lysin on the thermotropic phase behavior and vesicle morphology of dimyristoylphosphatidylcholine lipid bilayer model membranes . Differential scanning calorimetric, 31P nuclear magnetic resonance and Fourier transform infrared spectroscopic, and X-ray diffraction studies; Lohner K et al.; We investigated the effects of various concentrations of staphylococcal delta-lysin on the thermotropic phase behavior of large multilamellar dimyristoylphosphatidylcholine (DMPC) vesicles by differential scanning calorimetry (DSC), 31P nuclear magnetic resonance (NMR) and Fourier transform infrared (FTIR) spectroscopy, and X-ray diffraction . The DSC studies revealed that at all concentrations, the addition of delta-lysin progressively decreases the enthalpy of the pretransition of DMPC bilayers without significantly affecting its temperature or cooperativity . Similarly, the addition of smaller quantities of peptide has little effect on the temperature of the main phase transition of DMPC bilayers but does reduce the cooperativity and enthalpy of this transition somewhat . However, at higher peptide concentrations, a second phase transition with a slightly increased temperature and a markedly reduced cooperativity and enthalpy is also induced, and this latter phase transition resolves itself into two components at the highest peptide concentrations that are tested . Moreover, our 31P NMR spectroscopic studies reveal that at relatively low delta-lysin concentrations, essentially all of the phospholipid molecules produce spectra characteristic of the lamellar phase, whereas at the higher peptide concentrations, an increasing proportion exhibit an isotropic signal . Also, at the highest delta-lysin concentrations that are studied, the isotropic component of the 31P NMR spectrum also resolves itself into two components . At the highest peptide concentration that was tested, we are also able to effect a macroscopic separation of our sample into two fractions by centrifugation, a pellet containing relatively smaller amounts of delta-lysin and a supernatant containing larger amounts of peptide relative to the amount of lipid present . We are also able to show that the more cooperative phase transition detected calorimetrically, and the lamellar phase 31P NMR signal, arise from the pelleted material, while the less cooperative phase transition and the isotropic 31P NMR signal arise from the supernatant . In addition, we demonstrate by X-ray diffraction that the pelleted material corresponds to delta-lysin-containing large multilamellar vesicles and the supernatant to a mixture of delta-lysin-containing small unilamellar vesicles and discoidal particles . We also show by FTIR spectroscopy that delta-lysin exists predominantly in the alpha-helical conformation in aqueous solution or when interacting with DMPC, and that a large fraction of the peptide bonds undergo H-D exchange in D(2)O . However, upon interaction with DMPC, the fraction of exchangeable amide protons decreases . We also demonstrate by this technique that both of the phase transitions detected by DSC correspond to phospholipid hydrocarbon chain-melting phase transitions . Finally, we show by several techniques that the absolute concentrations of delta-lysin and the thermal history, as well as the lipid:peptide ratio, can affect the thermotropic phase behavior and morphology of peptide-lipid aggregates. J Hematother Stem Cell Res, 1999, 8 Suppl 1, S21 - 2 Anti-inflammatory aspects of Filgrastim and impact on IL-2 release; Hartung T; G-CSF has immunomodulatory effects of neutrophilic granulocytes and monocytes/macrophages . Two studies were done: one in normal volunteers and the other in HIV-infected patients plus their respective control donors to evaluate the effect of Filgrastim on cytokine responses . Filgrastim treatment of volunteers resulted in an anti-inflammatory cytokine response, when blood was stimulated ex vivo with the endotoxin lipopolysaccharide (LPS) . Similarly, in the presence of Filgrastim in vitro, the LPS-inducible release of proinflammatory cytokines was attenuated . Blood from HIV-infected patients at advanced stages of disease showed reduced interleukin (IL)-2 formation in response to staphylococcal exotoxin B (SEB), which was restored in the presence of Filgrastim. J Antimicrob Chemother, 1999 Nov, 44(5), 669 - 74 Foreign body infection: a new rat model for prophylaxis and treatment; Van Wijngaerden E et al.; A subcutaneous catheter model in the rat was developed that allowed the study of prevention and treatment strategies for foreign body infection . In contrast to earlier models, the foreign body was inoculated with a low inoculum of Staphylococcus epidermidis just before implantation, thus mimicking intraoperative contamination with skin flora . Reproducible infection of all catheters followed if no prophylaxis was given . However, foreign body infection could be prevented or treated with antibiotics such as teicoplanin, which was marginally effective, and rifampicin, which proved very effective. J Invest Dermatol, 1999 Dec, 113(6), 1082 - 9 1alpha,25-dihydroxycholecalciferol and cyclosporine suppress induction and promote resolution of psoriasis in human skin grafts transplanted on to SCID mice; Dam TN et al.; Accumulating evidence has emphasized the importance of immunocompetent cells in determining the psoriatic phenotype . We have investigated the effect of 1alpha,25-dihydroxycholecalciferol, the naturally occurring active form of vitamin D3, cyclosporine A, and interleukin-10 on the phenotype of human psoriatic skin xenotransplants . First, psoriatic skin transplants were injected with either 1alpha,25-dihydroxy- cholecalciferol, cyclosporine A, or interleukin-10 . Second, we determined the ability of autologous lymphocytes, activated in vitro using staphylococcal enterotoxin B and interleukin-2 and then exposed to either 1alpha, 25-dihydroxycholecalciferol or cyclosporine A, to induce psoriatic lesions if they were injected into the dermis of uninvolved skin grafts . We found that injections into transplanted psoriatic plaques of either 1alpha,25-dihydroxycholecalciferol or cyclosporine A, but not interleukin-10, resulted in a consistent reduction in the clinical and histologic score of psoriasis with remission towards uninvolved psoriatic skin . Injection of activated immunocytes into symptomless psoriatic skin grafts, changed the grafts towards plaque-type psoriasis with silvery scale, parakeratosis, elongated rete pegs, acanthosis, and dermal angiogenic reaction . In contrast, if activated immunocytes were exposed to 1alpha, 25-dihydroxycholecalciferol or cyclosporine A prior to injection, only minimal changes occurred . It was determined that neither staphylococcal enterotoxin B and interleukin-2 activation by itself, nor the drugs investigated, changed the CD4/CD8 ratio of activated (CD25 + ) cells . Our results are consistent with the hypothesis that psoriasis may be induced by activated T lymphocytes, and indicate that novel immunomodulatory drugs can serve to inhibit the pathogenetic ability of immunocytes in psoriasis. Neurol Neurochir Pol, 1999 May-Jun, 33(3), 699 - 702 {A rare case of intracranial hematoma complicating bacterial endocarditis}; Michalska-Krzanowska G et al.; Endocarditis is an infectious and inflammatory disease of cardiac valves with other coexisting symptoms affecting heart and other organs and systems . Patients with cardiac valves diseases are in the group of high risk . The authors present a case of successfully treated endocarditis affecting mitral valve prosthesis in the course of staphylococcus septicaemia complicated by intracranial haematoma. Mol Microbiol, 1999 Oct, 34(1), 10 - 23 The 213-amino-acid leucine-rich repeat region of the listeria monocytogenes InlB protein is sufficient for entry into mammalian cells, stimulation of PI 3-kinase and membrane ruffling; Braun L et al.; The Listeria monocytogenes InlB protein is a 630-amino-acid surface protein that mediates entry of the bacterium into a wide variety of cell types, including hepatocytes, fibroblasts and epithelial cells such as Vero, HEp-2 and HeLa cells . Invasion stimulates host proteins tyrosine phosphorylation, PI 3-kinase activity and rearrangements in the actin cytoskeleton . We previously showed that InlB is sufficient for entry of InlB-coated latex beads into cells and recent results indicate that purified InlB can stimulate PI 3-kinase activity and is thus the first bacterial agonist of this lipid kinase . In this study, we identified the region of InlB responsible for entry and stimulation of signal transduction events . Eight monoclonal antibodies directed against InlB were raised and, of those, five inhibited bacterial entry . These five antibodies recognized epitopes within the leucine-rich repeat (LRR) region and/or the inter-repeat (IR) region . InlB-staphylococcal protein A (SPA) fusion proteins and recombinant InlB derivatives were generated and tested for their capacity to mediate entry into cultured mammalian cells . All the InlB derivatives that carried the amino-terminal 213-amino-acid LRR region conferred invasiveness to the normally non-invasive bacterium L . innocua or to inert latex beads and the corresponding purified polypeptides inhibited bacterial entry . In addition, the 213-amino-acid LRR region was able to stimulate PI 3-kinase activity and changes in the actin cytoskeleton (membrane ruffling) . These properties were not detected with purified internalin, another invasion protein of L . monocytogenes that displays LRRs similar to those of InlB . Taken together, these results show that the first 213 amino acids of InlB are critical for its specific properties. Immunology, 1999 Oct, 98(2), 171 - 80 Induction of dendritic cell costimulator molecule expression is suppressed by T cells in the absence of antigen-specific signalling: role of cluster formation, CD40 and HLA-class II for dendritic cell activation; McLellan AD et al.; Full activation of T lymphocytes by dendritic cells (DC) during antigen presentation is known to require the interaction of several inducible receptor-ligand pairs . We have postulated that the reciprocal activation of DC by T lymphocytes is also important . Potential signalling molecules that might increase the stimulatory capacity of DC during antigen presentation to T lymphocytes were tested using an in vitro model . Fresh human blood DC were cocultured with CD4+ and CD8+ allogeneic or with autologous T lymphocytes plus Staphylococcus superantigen A (SEA) . Surprisingly, costimulator expression on DC cocultured with T lymphocytes was reduced in comparison to DC cultured alone . However, the minority (10-30%) of DC clustering with T lymphocytes showed antigen-specific up-regulation of the CD40, CD80 and CD86 costimulator molecules, whereas the non-clustered DC (70-90%) had less up-regulation than control DC cultured alone and did not respond to antigen-specific triggering . Monoclonal antibodies (mAb) to CD40 ligand (CD40L) and human leucocyte antigen (HLA)-DR, but not lymphocyte function-associated antigen-1 (LFA-1), LFA-3 or HLA-class I, significantly inhibited the T-lymphocyte induction of DC costimulator expression . Since HLA-class II, but not HLA-class I mAb, inhibited allogeneic T-lymphocyte-mediated activation of DC, CD4 T lymphocytes appear to be the main subset activating DC in the mixed lymphocyte reaction . Cross-linking of CD40, but not HLA-class II, up-regulated DC or B-cell costimulator expression . Although direct class II signalling does not appear to play a role in DC activation, antigen-specific T-cell recognition contributes via other mechanisms to regulate DC activation. Proteins, 1999 Nov 15, 37(3), 494 - 8 Kinetic characterization of the interaction of the Z-fragment of protein A with mouse-IgG3 in a volume in chemical space; Andersson K et al.; The kinetic rate parameters for the interaction between a single domain analogue of staphylococcal protein A (Z) and a mouse-IgG3 monoclonal antibody (MAb) were measured in Hepes buffer with different chemical additives . Five buffer ingredients (pH, NaCl, DMSO, EDTA, and KSCN) were varied simultaneously in 16 experiments following a statistical experimental plan . The 16 buffers thus spanned a volume in chemical space . A mathematical model, using data from the buffer composition, was developed and used to predict apparent kinetic parameters in five new buffers within the spanned volume . Association and dissociation parameters were measured in the new buffers, and these agreed with the predicted values, indicating that the model was valid within the spanned volume . The pattern of variation of the kinetic parameters in relation to buffer composition was different for association and dissociation, such that pH influenced both association and dissociation and NaCl influenced only dissociation . This indicated that the recognition mechanism (association) and the stability of the formed complex (dissociation) involve different binding forces, which can be further investigated by kinetic studies in systematically varied buffers. Mol Immunol, 1999 Aug, 36(11-12), 769 - 76 The murine clan V(H) III related 7183, J606 and S107 and DNA4 families commonly encode for binding to a bacterial B cell superantigen; Cary S et al.; Superantigens, by virtue of their unconventional binding interactions with Ag receptors, can simulate a large subset of mature lymphocytes in the repertoire . Recent studies have documented that in vivo exposure to the model bacterial B cell superantigen, Staphylococcal protein A (SpA), induces large scale effects on murine B-cell clonal selection by mechanism(s) that include deletion of supra-clonal sets . While the structural bases for the immunomodulatory properties of several T-cell superantigens have been well characterized, the requirements for murine Fab-binding of SpA remain incompletely defined . To investigate these structural requirements, a series of direct binding and inhibition studies were performed with a large panel of Moabs of diverse variable region gene usage . These studies confirm previous reports that superantigen binding is completely restricted to the products of clan V(H) III-related families, that include the small S107 and J606 families, and we also demonstrated that usage of the related small DNA4 family commonly correlates with weaker binding activity . Furthermore, our results document that genes from the largest clan V(H) III family, 7183, commonly encode for Fab-mediated binding of SpA, while antibodies from five other VH families, J558, Q52, Sm7, VH11 and VH12, did not display Fab-mediated SpA binding activity . By contributing to the essential foundation for understanding of the structural basis for binding interactions, these findings will aid interpretation of evolving observations regarding the clonal fates induced by in vivo B-cell superantigen exposure. Mayo Clin Proc, 1999 Dec, 74(12), 1227 - 30 Staphylococcus lugdunensis endocarditis: a complication of vasectomy? Fervenza FC, Contreras GE, Garratt KN, Steckelberg JM. Three cases of Staphylococcus lugdunensis endocarditis have been reported in patients with a history of vasectomy preceding the development of endocarditis . We describe a new case of a 39-year-old man who developed infective endocarditis due to S . lugdunensis after vasectomy . He was successfully treated with a 7-week course of intravenous antibiotics and subsequently underwent mitral valve reconstruction for severe mitral regurgitation . The present case further supports an association between vasectomy and S . lugdunensis endocarditis. Int Immunol, 1999 Dec, 11(12), 1873 - 80 Increased c-Fos/activator protein-1 confers resistance against anergy induction on antigen-specific T cell; Kawasaki H et al.; We have studied the contribution of c-Fos/activator protein-1 (AP-1) to antigen-specific T cell response with reference to T cell anergy by increasing c-Fos/AP-1 in vivo and in vitro . First, after injection of a high dose of staphylococcus enterotoxin B (SEB), clonal deletion of SEB-reactive V(beta)8(+) CD4 T cells occurred both in control B6 and H2-c-fos transgenic (fos) mice, whereas proliferation of T cells against SEB was profoundly depressed in B6 but not in fos mice . Second, the keyhole limpet hemocyanin-specific CD4 T(h)1 cell clone produced decreasing amounts of IL-2 in response to increasing amounts of concanavalin A (Con A) in vitro, whereas the decrease was less significant in the T(h)1 clones stably transfected with c-fos gene . Electrophoretic mobility shift assay with nuclear protein from the transformants showed that overexpression of the c-fos gene compensated the amounts of AP-1 in the nuclei of Con A-treated T(h)1 clones . Thus, increased c-Fos/AP-1 confers resistance against anergy induction on antigen-specific T cells. Dtsch Med Wochenschr, 1999 Oct 29, 124(43), 1267 - 70 {Heparin-induced thrombocytopenia type II: reexposure to heparin}; Matthies B et al.; HISTORY AND ADMISSION FINDINGS: At the age of 55 years a now 70-year-old man had his aortic valve replaced by a prosthetic (Bjork-Shiley) valve, and 11 years later a VDD pacemaker had been implanted . 18 months before the latest admission he had been hospitalized for treatment of staphylococcal endocarditis involving the aortic prothesis . At that time thrombocytopenia developed during heparin administration, diagnosed clinically and with the heparin-induced platelet activity (HIPA) test as type II heparin induced thrombocytopenia . His latest admission was for the diagnosis and treatment of peripheral arterial disease of the right leg (Fontaine stage IIb) . INVESTIGATIONS: Right popliteal and pedal pulses were not palpable . He was able to walk pain-free for only 70 m . Doppler sonography demonstrated an arm-leg index on the right of 0.7 . Angiography revealed marked stenosis in the right superficial femoral artery and a filiform stenosis in the right popliteal artery . TREATMENT AND COURSE: Both stenoses were relieved by percutaneous transluminal balloon angioplasty, in the course of which 5000 IU heparin were administered as a bolus intraarterially . Postoperative anticoagulation was maintained for 2 days with recombinant hirudin . There was no evidence of platelet reduction or heparin-induced antibodies despite the renewed infusion of heparin . CONCLUSION: Single re-administration of heparin in a patient who had developed a type II heparin-induced thrombocytopenia several years before does not necessarily lead to a booster of antibodies and thus to a reduction of platelets in the peripheral blood . It is a moot point whether the course in this case was an exception or the rule. Biosci Biotechnol Biochem, 1999 Oct, 63(10), 1828 - 30 Construction of a LukS-PV mutant of a staphylococcal Panton-Valentine leukocidin component having a high LukS-like function; Shimatani A et al.; A 2-residue (D12I13) segment of LukS of a staphylococcal leukocidin component is an essential region for the hemolytic function of LukS towards rabbit erythrocytes in the presence of LukF . Here, we report that insertion of D, I, or AA residue(s) between A11 and E12 residues of LukS-PV, in which the 2-residue D12I13 segment in LukS was absent, confers the full LukS function on LukS-PV, which has only 4% hemolytic activity of that of LukS towards rabbit erythrocytes. J Immunol, 1999 Dec 15, 163(12), 6686 - 93 Staphylococcal enterotoxin H displays unique MHC class II-binding properties; Nilsson H et al.; Staphylococcal enterotoxin H (SEH) has been described as a superantigen by sequence homology with the SEA subfamily and briefly characterized for its in vivo activity . In this study, we demonstrate that SEH is a potent T cell mitogen and inducer of T cell cytotoxicity that possesses unique MHC class II-binding properties . The apparent affinity of SEH for MHC class II molecules is the highest affinity ever measured for a staphylococcal enterotoxin (Bmax1/2 approximately 0.5 nM for MHC class II expressed on Raji cells) . An excess of SEA or SEAF47A, which has reduced binding to the MHC class II alpha-chain, is able to compete for binding of SEH to MHC class II, indicating an overlap in the binding sites at the MHC class II beta-chain . The binding of SEH to MHC class II is like SEA, SED, and SEE dependent on the presence of zinc ions . However, SEH, in contrast to SEA, binds to the alanine-substituted DR1 molecule, betaH81A, believed to have impaired zinc-bridging capacity . Furthermore, alanine substitution of residues D167, D203, and D208 in SEH decreases the affinity for MHC class II as well as its in vitro potency . Together, this indicates an MHC class II binding site on SEH with a different topology as compared with SEA . These unique binding properties will be beneficial for SEH to overcome MHC class II isotype variability and polymorphism as well as to allow an effective presentation on APCs also at low MHC class II surface expression. Eur J Pediatr Surg, 1999 Oct, 9(5), 343 - 6 Association of inflammatory pseudotumor of the liver and Papillon-Lefevre syndrome--case report; Czauderna P et al.; A case of hepatic inflammatory pseudotumor mimicking malignancy in a 4-year-old girl with the Papillon-Lefevre syndrome (PLS) is reported . Only recently, an association between this inherited syndrome and liver abscesses has been found . Its possible pathogenesis is discussed and immunologic defects resulting from the Papillon-Lefevre syndrome are presented . The development of inflammatory pseudotumor of the liver might be caused by immunologic disturbances and staphylococcal infection . The picture of the hepatic tumor on imaging in patients with PLS should be attributed rather to inflammatory than neoplastic process. Immunol Res, 1999, 20(2), 163 - 73 Immune response to staphylococcal superantigens; Krakauer T; Staphylococcal exotoxins, staphylococcal enterotoxins A-E (SEA-SEE), and toxic shock syndrome toxin- (TSST-1) are potent activators of the immune system and cause a variety of diseases in humans, ranging from food poisoning to shock . These toxins are called superantigens because of their ability to polyclonally activate T cells at picromolar concentrations . Superantigens bind to both MHC class II molecules and specific Vbeta regions of the T cell receptor, leading to the activation of both antigen-presenting cells and T lymphocytes . These interactions lead to excessive production of proinflammatory cytokines and T cell proliferation, causing clinical symptoms that include fever, hypotension, and shock . Recent studies suggest that staphylococcal superantigens may also be involved in the pathogenesis of arthritis and other autoimmune disorders . This review summarizes the in vitro and in vivo effects of staphylococcal enterotoxins and TSST-1, recent progress with the use of transgenic knockout mice to identify key mediators and receptors involved in superantigen-induced shock, and therapeutic agents to mitigate the toxic effects of staphylococcal superantigens. J Hepatol, 1999 Nov, 31(5), 815 - 24 Interleukin-10 inhibits hepatic injury and tumor necrosis factor-alpha and interferon-gamma mRNA expression induced by staphylococcal enterotoxin B or lipopolysaccharide in galactosamine-sensitized mice; Nagaki M et al.; BACKGROUND/AIMS: Proinflammatory cytokines including tumor necrosis factor alpha (TNF-alpha) and interferon gamma (IFN-gamma) play a critical role in the pathogenesis of liver injury induced by lipopolysaccharide (LPS) or staphylococcal enterotoxin B (SEB) in D-galactosamine (GalN)-sensitized mice . The aim of this study was to examine the ability of interleukin-10 (IL-10), a recently characterized, highly potent anti-inflammatory mediator, to protect sensitized mice against hepatotoxicity induced by SEB or LPS . METHODS: IL-10 was injected at various concentrations into BALB/c mice treated by GalN/SEB or GalN/LPS . Liver injury was assessed biochemically and histologically . Serum levels of TNF-alpha and IFN-gamma were measured and the expressions of TNF-alpha and IFN-gamma mRNA in the liver and spleen were determined by reverse-transcription polymerase chain reaction . RESULTS: Treatment with IL-10 markedly reduced serum transaminase activities in a dose-dependent manner and reduced hemorrhagic liver damage in sensitized mice exposed to either toxin . IL-10 also inhibited increases in serum TNF-alpha and IFN-gamma concentrations with either toxin . Treatment with IL-10 significantly reduced TNF-alpha mRNA and IFN-gamma mRNA expression in the liver and spleen after administration of either toxin to sensitized mice . CONCLUSIONS: These findings suggest that IL-10 is capable of regulating both T cell- and macrophage-mediated hepatic injury in vivo and that this cytokine might be useful in the treatment of acute liver failure. Ann Rheum Dis, 1999 Nov, 58 Suppl 1, I73 - 81 PEGylated recombinant human soluble tumour necrosis factor receptor type I (r-Hu-sTNF-RI): novel high affinity TNF receptor designed for chronic inflammatory diseases; Edwards CK 3rd; The proinflammatory cytokine, tumour necrosis factor alpha (TNFalpha) has been shown to play a pivotal part in mediating acute and chronic inflammation . The activities of TNFalpha are modulated by the proteolytic shedding of the soluble extracellular domains of the two TNF receptors, p55 sTNF-RI and p75 sTNF-RII . Amgen Inc has cloned and expressed a recombinant form of a natural inhibitor of TNFalpha, referred to as recombinant human soluble TNF receptor type I (r-Hu-sTNF-RI, sTNF-RI) . sTNF-RI is an E coli recombinant, monomeric form of the soluble TNF-type I receptor . A high molecular weight polyethylene glycol (PEG) molecule is attached at the N-terminus position to form the molecule intended for clinical evaluations (PEG sTNF-RI) . Preclinical studies to date demonstrate that PEG sTNF-RI is efficacious in rodent models of chronic inflammatory disease including rheumatoid arthritis and Crohn's disease at doses as low as 0.3 mg/kg given every other day . This dose results in plasma concentrations of 0.3 to 0.5 microg/ml . Higher doses with correspondingly higher plasma concentrations yield higher efficacy . It has also demonstrated efficacy in E coli lipopolysaccharide, and Staphylococcus enterotoxin B mediated models of acute inflammation in rodents and primates . Pharmacokinetic studies in mice, rats, cynomolgus monkeys, baboons, and chimpanzees have been conducted with PEG sTNF-RI . Absorption from a subcutaneous dose was slow, with the time to reach maximal plasma concentrations of 24-48 hours in rats, and in monkeys, and 3-29 hours in chimpanzees . The initial volume of distribution of PEG sTNF-RI was essentially equivalent to that of plasma (40 ml/kg) . This suggests the protein does not appear to extensively distribute from the systemic circulation with a volume of distribution at steady state (Vss) less than 200 ml/kg in all species studied . These results are consistent with previous experience with PEGylated proteins in which PEGylation decreases both the rate of absorption and the plasma clearance of human recombinant proteins in animals and humans . The use of a PEG molecule will probably provide a more advantageous dosing schedule (that is, less frequent dosing) for the patient compared with a non-PEG sTNF-RI. Res Microbiol, 1999 Oct, 150(8), 531 - 41 Distribution of Staphylococcus sciuri subspecies among human clinical specimens, and profile of antibiotic resistance; Marsou R et al.; The distribution of three subspecies comprising Staphylococcus sciuri was determined for a collection of 30 clinical isolates originating from Morocco, the United Kingdom, and France . The sources of these isolates were principally wounds, skin, and soft tissue infections . At the species level, the isolates were identified according to biochemical characteristics and at the subspecies level by the ribotyping technique . PCR analysis performed with the 16S-23S ribosomal DNA intergenic spacer was less powerful for subspecies differentiation . S . sciuri subsp . sciuri was the most frequent subspecies (21 isolates) found in the collection, whereas S . sciuri subsp . rodentium (seven isolates) and S . sciuri subsp . carnaticus (two isolates) were less common . mecA or a mecA-related gene was detected by PCR and Southern blot in all 30 S . sciuri isolates, supporting the suggestion that S . sciuri species are the natural reservoir of the mecA gene . While the linA/linA' gene coding for lincomycin resistance was present in five isolates, an uncharacterized gene for this resistance was suspected in seventeen other isolates. Khirurgiia (Mosk), 1999, (10), 29 - 34 {Antimicrobial chemotherapy in patients with pyo-septic diseases in intensive care units}; Iakovlev VP et al.; The analysis of the treatment of 900 patients with large festered wounds of various genesis and location for the period from 1973 to 1998 years in the intensive care department has shown, that infection of respiratory ways is encountered in 30% of cases (in patients with nonsporeforming anaerobic bacteria--in 11-12%), bacteriuria--in 70-80%, bacteriamia--in 75% of patients with sepsis . In acute pyogenous diseases of soft tissues microbes from the wounds in monoculture were isolated out in 83.3% of cases, associations of gram-positive and gram-negative bacteria--in 16.7%, in chronic pyogenous diseases of soft tissues--in 60 and 40% of cases, respectively . In sepsis associations of gram-positive and gram-negative microbes were isolated in 55.6% of cases . Most often (91%) pathogenic staphylococcus was found in hemocultures . Uring in 62% of cases contained association of gram-positive and gram-negative microorganisms, in sputum gram-positive microflora in monoculture (69%) prevailed . In the group of patients with peritonitis, phlegmon of the anterior abdominal wall, diabetic phlegmon and gangrene, crush syndrome the association of anaerobic and aerobic microflora (from 57.1 to 98.8%) prevailed in the wounds . Application of up-to-date antimicrobial means in the intensive care unit resulted in a decrease of mortality rate in sepsis and complicated course of wound infection up to 23%, and in anaerobic nonsporeforming infection--up to 15%. Am J Nephrol, 1999, 19(5), 605 - 8 Staphylococcus lugdunensis pulmonary valve endocarditis in a patient on chronic hemodialysis; Kamaraju S et al.; We describe a case of Staphylococcus lugdunensis pulmonary valve endocarditis in a 65-year-old woman on chronic hemodialysis and provide a review of previously reported cases . The patient presented with fever and altered mental status, but had no other localizing symptoms or signs; coagulase-negative staphylococcus (subsequently identified as S . lugdunensis) was isolated from two sets of blood cultures . Transthoracic and transesophageal echocardiograms showed a large (2.3 x 3.1 cm) vegetation on the pulmonary valve with moderate valvular insufficiency . The patient was treated with 6 weeks of antibiotic therapy and is stable 4 months following the completion of therapy; no surgical intervention was performed . Of the 28 previously reported cases of S . lugdunensis endocarditis, only 1 had previously survived with medical therapy alone . This is the 3rd case report of S . lugdunensis endocarditis in a patient on hemodialysis; the presumed portal of entry in this and previously reported cases was the vascular access device . Endocarditis due to this organism is characterized by a high mortality, rapid tissue destruction, and a predilection for native valves . Because the clinical outcome is much more favorable with valvular replacement, speciation of the organism assumes great importance in defining the therapeutic approach . Copyright Heart, 1999 Dec, 82(6), 714 - 20 Long term follow up of prosthetic valve endocarditis: what characteristics identify patients who were treated successfully with antibiotics alone? Truninger K, Attenhofer Jost CH, Seifert B, Vogt PR, Follath F, Schaffner A, Jenni R. OBJECTIVE: To identify predictors for the safe use of antibiotic treatment without reoperation in patients with prosthetic valve endocarditis . SETTING: Retrospective study in a tertiary care centre . SUBJECTS AND DESIGN: All 49 episodes of definite prosthetic valve endocarditis (Duke criteria) diagnosed at one institution between 1980 to 1997 were analysed . Ten episodes (20%) were treated with antibiotics only (antibiotic group) and 39 episodes (80%) with combined antibiotic and surgical treatment (surgery group) . The analysis included detailed study of hospital records and data on long term follow up which were obtained in all patients by a questionnaire or telephone contact with physician or patient . The length of follow up (mean (SD)) was 41 (32) months in the antibiotic group and 45 (24) months in the surgery group (NS) . Long term survival was estimated by the Kaplan-Meier method and compared by the log-rank test . RESULTS: There was no significant difference in age, history of previous endocarditis, number of previous heart operations, vegetations, emboli, type of prosthesis, or percentage of early prosthetic valve endocarditis and positive blood cultures between the two groups . In the antibiotic group, there were more enterococcal (50%; p = 0.005) and in the surgery group more staphylococcal infections (55%; p = 0.048) . Annular abscesses (p < 0 . 0001) and aortoventricular dehiscence (p = 0.02) were more common in the surgery group . No patient in the antibiotic group had heart failure . Long term follow up showed no significant difference between the surgery and antibiotic groups regarding late mortality (14% v 18%) and five year rates of recurrent endocarditis (14% v 16%), event related mortality (14% v 3%, log-rank test), and the need for reoperation (14% v 19%; log-rank test) . The only patient with conservatively treated staphylococcal prosthetic valve endocarditis died after reoperation for recurrence . CONCLUSIONS: Haemodynamically stable patients with non-staphylococcal prosthetic valve endocarditis who are carefully supervised can be treated with antibiotics alone without an increased rate of reinfection, reoperation, or death. Am J Respir Cell Mol Biol, 1999 Dec, 21(6), 675 - 83 Role of T cells in bronchoalveolar space in the development of interstitial pneumonia induced by superantigen in autoimmune-prone mice; Fujiki M et al.; To study the mechanisms underlying the development of interstitial pneumonia in autoimmune disease, we analyzed bronchoalveolar lavage fluid (BALF) in an animal model of interstitial pneumonia in which an intratracheal instillation of staphylococcal enterotoxin B (SEB) induced interstitial pneumonia in autoimmune-prone mice . Increases in the numbers of total cells, macrophages, lymphocytes, and neutrophils were observed in BALF from SEB-treated MRL +/+ mice, and peaked at 3 d after SEB administration (Day 3) . Flow cytometric analyses revealed increases in SEB-reactive Vbeta8(+) T cells, indicating that SEB-reactive cells play an important role in bronchoalveolar space . The expressions of tumor necrosis factor (TNF)-alpha, interferon (IFN)-gamma, JE/monocyte chemoattractant protein-1, regulated on activation, normal T cells expressed and secreted, and KC/gro messenger RNA (mRNA) in BALF cells from SEB-treated mice peaked at Day 3 . Increased expression of TNF-alpha mRNA was observed mainly in macrophages and CD8(+) T cells, and the increase in IFN-gamma mRNA was observed mainly in CD8(+) T cells in BALF at Day 3 . The expression of platelet-derived growth factor mRNA was very weak at Day 3 but strongly expressed at Day 14 . An immunosuppressant, FK506, but not corticosteroid, suppressed SEB-induced T-cell expansion in BALF as well as increased cytokine and chemokine production in the bronchoalveolar space of SEB-treated mice . Histologically, FK506 but not corticosteroid significantly reduced both the cell infiltration to alveolar septal walls and the synthesis of pulmonary collagen fibers . Further, transfer of T cells of MRL +/+ mice with SEB into SCID mice gave rise to interstitial pneumonia . These results suggest that superantigen-reactive T cells in the bronchoalveolar space may trigger the development of interstitial pneumonia in this model. Biochemistry, 1999 Oct 26, 38(43), 14271 - 8 Carboxylate groups on the manganese-stabilizing protein are required for its efficient binding to photosystem II; Frankel LK et al.; The effects of the modification of carboxylate groups on the manganese-stabilizing protein of photosystem II were investigated . Carboxylate groups (including possibly the C-terminus) on the manganese-stabilizing protein were modified with glycine methyl ester in a reaction facilitated by 1-ethyl-3-{3-(dimethylamino)propyl}carbodiimide . The manganese-stabilizing protein that was modified while associated with NaCl-washed photosystem II membranes contained 1-2 modified carboxylates, whereas the protein that was modified while free in solution contained 4 modified carboxylates . Both types of modified protein could reconstitute oxygen evolution at high manganese-stabilizing protein to photosystem II reaction center ratios . However, the protein that had been modified in solution exhibited a dramatically altered binding affinity for photosystem II . No such alteration in binding affinity was observed for the protein that had been modified while associated with the photosystem . Mapping of the sites of modification was carried out by trypsin and Staphylococcus V8 protease digestion of the modified proteins and analysis by matrix-assisted laser desorption/ionization mass spectrometry . These studies indicated that the domains (157)D-(168)D and (212)E-(247)Q (C-terminus) are labeled only when the manganese-stabilizing protein is modified in solution . Modified carboxylates in these domains are responsible for the altered binding affinity of this protein for the photosystem. J Food Prot, 1999 Nov, 62(11), 1350 - 3 Emesis in the shrew mouse (Suncus murinus) induced by peroral and intraperitoneal administration of staphylococcal enterotoxin A; Hu DL et al.; Peroral and intraperitoneal administration of staphylococcal enterotoxin A (SEA) to Suncus murinus elicited an emetic response . The 50% emetic dose of SEA by peroral administration was found to be 32 microg per kg of body weight, whereas that by intraperitoneal administration was 3 microg per kg . Multiple emetic responses occurred 70 to 108 min after peroral administration of an emetic dose of SEA . Similar responses occurred 65 to 102 min after intraperitoneal injection of an emetic dose of SEA . No significant difference in vomiting was observed between male and female animals . Anti-SEA serum neutralized SEA-induced emesis in S . murinus . These findings indicate that S . murinus may serve as a suitable animal model to study the enterotoxigenicity of SEA. Proc Natl Acad Sci U S A, 1999 Nov 23, 96(24), 13944 - 9 Trafficking of spontaneously endocytosed MHC proteins; Chiu I et al.; Class I MHC protein primarily presents endogenous antigen but also may present exogenous antigen . Here, we investigated the intracellular pathway of spontaneously internalized class I MHC protein by confocal microscopy . beta(2)-microglobulin (beta(2)m), labeled with a single fluorophore, was exchanged at the surface of B cell transfectants to specifically mark cell surface and endocytosed class I MHC protein . Intracellular beta(2)m colocalized with fluorophore-conjugated transferrin, implying that class I MHC protein endocytosed into early endosomes . These endosomes containing fluorescent beta(2)m were found close to or within the Golgi apparatus, marked by fluorescent ceramide . Even after 24 hr of incubation, very little fluorescent beta(2)m was found in intracellular organelles stained by DiOC(6), marking the endoplasmic reticulum, or fluorophore-conjugated low density lipoprotein, marking late endosomes and lysosomes . Fluorophore-conjugated superantigens (staphylococcal enterotoxin A and B), presumed to enter cells bound to class II MHC protein, also were found to endocytose into beta(2)m-containing early endosomes . Staining with mAb and use of transfectants expressing MHC protein attached to green fluorescent protein confirmed the presence of intracellular compartments rich in both class I and II MHC protein and demonstrated that class I and II MHC protein also colocalize in discrete microdomains at the cell surface . These cell surface microdomains also contained transferrin receptor and often were juxtaposed to cholesterol-rich lipid rafts . Thus, class I and II MHC protein meet in microdomains of the plasma membrane and endocytose into early endosomes, where both may acquire and present exogenous antigen. Proc Natl Acad Sci U S A, 1999 Nov 23, 96(24), 13932 - 7 A transgenic mouse model to analyze CD8(+) effector T cell differentiation in vivo; Manjunath N et al.; Antigen-specific effector T cells are prerequisite to immune protection, but because of the lack of effector cell-specific markers, their generation and differentiation has been difficult to study . We report that effector cells are highly enriched in a T cell subset that can be specifically identified in transgenic (T-GFP) mice expressing green fluorescent protein (GFP) under control of the murine CD4 promoter and proximal enhancer . Consistent with previous studies of these transcriptional control elements, GFP was strongly and specifically expressed in nearly all resting and short-term activated CD4(+) and CD8(+) T cells . However, when T-GFP mice were challenged with vaccinia virus, allogeneic tumor cells, or staphylococcal enterotoxin A, the cytotoxic and IFN-gamma-producing T cells lost GFP expression . Upon T cell receptor (TCR) ligation by alphaCD3, sorted GFP(+) cells fluxed calcium and proliferated vigorously . In contrast, GFP(-) effector cells showed a diminished calcium flux and did not proliferate . Instead, they underwent apoptosis unless supplied with exogenous IL-2 . By reverse transcription-PCR analysis, the GFP(-) cells up-regulated the pro-apoptotic molecule, Fas-L, and down-regulated gene expression of the proximal TCR signaling molecule, CD3zeta, and c-jun, a component of the AP-1 transcription factor . Thus, differential regulation of TCR signaling may explain the divergent responses of naive and effector T cells to antigen stimulation. Infect Immun, 1999 Dec, 67(12), 6611 - 8 Production of tumor necrosis factor alpha in human T lymphocytes by staphylococcal enterotoxin B correlates with toxin-induced proliferation and is regulated through protein kinase C; Yan Z et al.; The superantigen staphylococcal enterotoxin B (SEB) simultaneously binds both the major histocompatibility complex (MHC) class II receptor on monocytes and the T-cell receptor (TCR) on T lymphocytes, resulting in a range of cell responses including induction of tumor necrosis factor alpha (TNF-alpha) . In this study, we have used mixed cultures of human peripheral blood monocytes and lymphocytes to investigate biochemical events controlling SEB induction of TNF-alpha . TNF-alpha production induced by SEB in mixed cultures is more closely associated with T cells than with monocytes: (i) a TCR-binding-site mutant of SEB (N23F) is less active in TNF-alpha induction than an MHC class II receptor-binding-site mutant (F44R), and (ii) flow cytometric analysis indicated that SEB induced TNF-alpha production in T cells but not in monocytes . Pretreatment of cells with inhibitors of signal transduction pathways was employed to further define events in SEB-induced TNF-alpha production . Neither protein kinase A inhibitors nor two protein tyrosine kinase inhibitors altered SEB-induced TNF-alpha production . In contrast, SEB induced protein kinase C (PKC) translocation, and pretreatment of cultures with inhibitors of PKC blocked TNF-alpha induction . Alteration of levels of diacylglycerol (DAG), an activator of PKC, by treatment with inhibitors of phospholipase C or DAG kinase also altered SEB-induced TNF-alpha production . These data suggest that PKC activation plays a critical role in SEB-induced TNF-alpha production in human T cells. FEBS Lett, 1999 Nov 19, 461(3), 280 - 6 Discoupling the Ca(2+)-activation from the pore-forming function of the bi-component Panton-Valentine leucocidin in human PMNs; Baba Moussa L et al.; The consecutive cell activation, including Ca(2+)-channel opening, and pore formation leading to human neutrophil lysis were the two functions of the staphylococcal Panton-Valentine leucocidin attempted to be discoupled by site-directed mutagenesis . In a first approach consisting in deletions of the cytoplasmic extremity of the transmembranous domain, we produced a LukF-PV DeltaSer125-Leu128 with a slightly reduced Ca(2+) induction but with a significantly lowered lytic activity when combined with its synergistic protein LukS-PV . The second approach consisted in the modification of charges and/or introduction of a steric hindrance inside the pore, which also led to interesting mutated proteins: LukF-PV G131D, G131W and G130D . The latter had an intact Ca(2+) induction ability while the lytic one was 20-fold diminished . Binding properties and intrinsic pore diameters of these discoupled toxins remained comparable to the wild-type protein . The mutated proteins promoted interleukin-8 secretion, but they were rather inactive in an experimental model . New insights are brought concerning the role of the two functions in the virulence of this bi-component leucotoxin. Mol Cell Biol, 1999 Dec, 19(12), 8604 - 15 Cytosolic delivery of granzyme B by bacterial toxins: evidence that endosomal disruption, in addition to transmembrane pore formation, is an important function of perforin; Browne KA et al.; Granule-mediated cell killing by cytotoxic lymphocytes requires the combined actions of a membranolytic protein, perforin, and granule-associated granzymes, but the mechanism by which they jointly kill cells is poorly understood . We have tested a series of membrane-disruptive agents including bacterial pore-forming toxins and hemolytic complement for their ability to replace perforin in facilitating granzyme B-mediated cell death . As with perforin, low concentrations of streptolysin O and pneumolysin (causing <10% (51)Cr release) permitted granzyme B-dependent apoptosis of Jurkat and Yac-1 cells, but staphylococcal alpha-toxin and complement were ineffective, regardless of concentration . The ensuing nuclear apoptotic damage was caspase dependent and included cleavage of poly(ADP-ribose) polymerase, suggesting a mode of action similar to that of perforin . The plasma membrane lesions formed at low dose by perforin, pneumolysin, and streptolysin did not permit diffusion of fluorescein-labeled proteins as small as 8 kDa into the cell, indicating that large membrane defects are not necessary for granzymes (32 to 65 kDa) to enter the cytosol and induce apoptosis . The endosomolytic toxin, listeriolysin O, also effected granzyme B-mediated cell death at concentrations which produced no appreciable cell membrane damage . Cells pretreated with inhibitors of endosomal trafficking such as brefeldin A took up granzyme B normally but demonstrated seriously impaired nuclear targeting of granzyme B when perforin was also added, indicating that an important role of perforin is to disrupt vesicular protein trafficking . Surprisingly, cells exposed to granzyme B with perforin concentrations that produced nearly maximal (51)Cr release (1,600 U/ml) also underwent apoptosis despite excluding a 8-kDa fluorescein-labeled protein marker . Only at concentrations of >4,000 U/ml were perforin pores demonstrably large enough to account for transmembrane diffusion of granzyme B . We conclude that pore formation may allow granzyme B direct cytosolic access only when perforin is delivered at very high concentrations, while perforin's ability to disrupt endosomal trafficking may be crucial when it is present at lower concentrations or in killing cells that efficiently repair perforin pores. Rev Esp Cardiol, 1999 Oct, 52(10), 869 - 71 {Aortic periprosthetic abscess with extension to both atria}; Morillas PJ et al.; The periprosthetic abscess due to infective endocarditis constitutes a severe complication of an aortic valve replacement, causing high mortality, despite combined medical and surgical treatment, especially in "early" endocarditis . Transthoracic echocardiography, and especially transesophageal study, is the election procedure for a non invasive diagnosis of vegetation and local complications . We report the aggressive and fulminant case of a 43 year old woman with aortic periprosthetic abscess and the extension to both auricles, due to Staphylococcus epidermidis. Eur J Immunol, 1999 Nov, 29(11), 3516 - 26 CD40 associates with the MHC class II molecules on human B cells; Leveille C et al.; The MHC class II and CD40 molecules are two major components of the immune system that are involved in cell-cell interactions and signal transduction . Data obtained in the course of the present investigation show that these two molecules are physically associated on the surface of various human B cell lines and on normal tonsilar B cells . The CD40 / MHC class II complexes were not detected on the germinal center B cell line Ramos . However, stimulation of these cells via CD40 or MHC class II triggered their association, suggesting that the formation of the complex is related to the activation status of the cells . The formation of these complexes did not alter the interaction of MHC class II molecules with one of their natural ligands, the staphylococcal enterotoxin A (SEA), as evidenced by the ability of SEA to bind MHC class II / CD40 complexes . Cross-linking of MHC class II or CD40 molecules leads to the association as well as the co-association of both molecules to the NP-49-insoluble cellular matrix . Such association allowed us to demonstrate that only a fraction of these molecules can be physically associated on the cell surface . Based on previous observations and those presented here, it is highly possible that the CD40 / MHC class II complexes may have an important role in signal(s) induced via both molecules and during T / B cells interactions. Protein Eng, 1999 Oct, 12(10), 873 - 8 Affinity maturation of a Taq DNA polymerase specific affibody by helix shuffling; Gunneriusson E et al.; The possibility of increasing the affinity of a Taq DNA polymerase specific binding protein (affibody) was investigated by an alpha-helix shuffling strategy . The primary affibody was from a naive combinatorial library of the three-helix bundle Z domain derived from staphylococcal protein A . A hierarchical library was constructed through selective re-randomization of six amino acid positions in one of the two alpha-helices of the domain, making up the Taq DNA polymerase binding surface . After selections using monovalent phage display technology, second generation variants were identified having affinities (K(D)) for Taq DNA polymerase in the range of 30-50 nM as determined by biosensor technology . Analysis of binding data indicated that the increases in affinity were predominantly due to decreased dissociation rate kinetics . Interestingly, the affinities observed for the second generation Taq DNA polymerase specific affibodies are of similar strength as the affinity between the original protein A domain and the Fc domain of human immunoglobulin G . Further, the possibilities of increasing the apparent affinity through multimerization of affibodies was demonstrated for a dimeric version of one of the second generation affibodies, constructed by head-to-tail gene fusion . As compared with its monomeric counterpart, the binding to sensor chip immobilized Taq DNA polymerase was characterized by a threefold higher apparent affinity, due to slower off-rate kinetics . The results show that the binding specificity of the protein A domain can be re-directed to an entirely different target, without loss of binding strength. Biochemistry, 1999 Nov 9, 38(45), 14897 - 905 An engineered minidomain containing an elastin turn exhibits a reversible temperature-induced IgG binding; Reiersen H et al.; A two-helix version of the triple alpha-helical staphylococcal Protein A, previously shown to retain the Fc binding properties of protein A, has been engineered to contain an elastin sequence, GVPGVG, within the inter-helix turn . The original type I beta-turn was replaced with a beta-turn from the muscle protein elastin, which has an inverse temperature-induced folding transition . These "elastin mutants" had lost their helical structure, as measured by circular dichroism (CD), and exhibited a lower stability than the wild-type domains (T(m) reduced by about 48 degrees C) in 30% trifluoroethanol . For the wild-type domains, the amount of alpha-helix and the binding affinity for Fc decreased as the temperature was increased . In contrast, although the starting affinity was lower for the disulfide elastin-turn mutant, it exhibited a 21-fold improvement in affinity over the same temperature range . The melting curve for the elastin-turn minidomain showed cooperative behavior, as measured by the increase in CD-amplitude at 222 nm . The observed CD behavior is consistent with the formation of a type I beta-turn, exhibiting similar DeltaH and DeltaS values to those seen previously for short elastin peptides {Reiersen, H., Clarke, A . R., and Rees, A . R . (1998) J . Mol . Biol . 283, 255-264}, and accounting for the increase in on-rate . This demonstrates that, when inserted into a stable globular protein, short elastin sequences have the ability to modify local structure and activity, by operating as temperature modulated switches. CLAO J, 1999 Oct, 25(4), 228 - 32 Microbiological evaluation of contact lenses and contact lens disinfection solutions in an asymptomatic population and in medical personnel; Kozer-Bilgin L et al.; PURPOSE: To compare the contamination of contact lenses and contact lens disinfection solutions in a population of contact lenses wearers comprised of medical personnel and non-medical subjects . METHODS: Forty-six medical personnel working in an infectious environment and 35 non-medical subjects were enrolled in the study . Contact lenses and contact lens disinfecting solutions were cultured and potential pathogens were isolated . RESULTS: The frequency of isolation of pathogens from the contact lenses of medical personnel was significantly higher than from non-medical subjects . There was no growth in both the contact lens and the disinfecting solution cultures in 56.5% of the subjects in the medical personnel group, and in 57.1% of non-medical group of subjects . Both the contact lens and disinfecting solution cultures were positive in 21.7% of the medical personnel group, while in the non-medical group, only 2.9% of subjects had positive cultures . CONCLUSIONS: Microbacterial contamination of contact lenses and disinfection solution was more frequent in the medical personnel group compared with non-medical subjects . In the medical personnel group, the most frequently isolated microorganism was Staphylococcus albus (19.8%), which is seen in hospital infections . Because soft and rigid gas permeable contact lens usage was approximately equal, we conclude that medical personnel are more prone to contamination. Br J Haematol, 1999 Jun, 105(4), 1086 - 91 Differentiation of autoimmune thrombocytopenia from thrombocytopenia associated with immune complex disease: systemic lupus erythematosus, hepatitis-cirrhosis, and HIV-1 infection by platelet and serum immunological measurements; Samuel H et al.; A method and approach are described to differentiate classic autoimmune thrombocytopenia (ATP) from immune complex-associated thrombocytopenia in systemic lupus (SLE), hepatitis/chronic liver disease (LIV-ITP) and HIV-1 related thrombocytopenia (HIV-1-ITP) . The platelet immunologic profile of IgG, C3C4 and IgM was measured with a solid-phase ELISA, employing 125I-staphylococcal protein A to detect indicator antibody binding . Polyethylene glycol was employed to precipitate immune complexes (PEG-IC) . Platelet-associated IgG (PAIgG) was 2.8-, 5.6- and 5.8-fold higher in SLE, LIV-ITP and HIV-1-ITP patients respectively compared to ATP patients: platelet C3C4 was 3.2-, 4.8- and 4.5-fold higher respectively; platelet IgM was 2.2-, 3.7- and 3.8-fold higher respectively; serum PEG-IC levels were 4.2-, 4.8- and 2.1-fold higher respectively . With all parameters measured, there was no overlap between the 75th percentile for ATP patients and the 25th percentile for all three cohorts . The likelihood of having a platelet C3C4 level higher than the highest ATP level was 69% for SLE, 90% for LIV-ITP and 94% for HIV-1-ITP respectively; with PEG-IC measurements the likelihood was 83%, 100% and 100% respectively . Serum IgG, C3, C4, IgM and PEG-IC were examined for a possible relationship with platelet measurements . Except for a positive correlation between serum and platelet IgM in ATP, r = 0.5, P < 0.04, there was no positive correlation with any of the parameters measured . An inverse correlation was noted between PEG-IC level and platelet C3C4 in SLE, r = 0.7, P < 0.04 . Thus platelet immunologic profile and serum PEG-IC level measurements differentiated classic ATP from immune complex-associated thrombocytopenias (SLE, LIV-ITP, HIV-1-ITP) . Except for IgM measurements in ATP, platelet measurements could not be attributed to their respective serum concentration. Eur Arch Otorhinolaryngol, 1999, 256(9), 439 - 41 Necrotizing otitis externa caused by Staphylococcus epidermidis; Soldati D et al.; We present a case of malignant necrotizing otitis externa (MNOE) caused by Staphylococcus epidermidis, which is usually a non-pathogenic microorganism . The patient is an otherwise healthy, nondiabetic 58-year-old white man . Contributory history began in 1994 after surgery for bilateral exostoses of the external auditory canals . Between April 1994 and May 1998 persistent otalgia occurred, with progressive mixed hearing losses, purulent discharge from both ears, spontaneous perforations of the tympanic membranes and ulceration of canal wall skin . From the beginning, Staph . epidermidis was isolated in all but one culture, but was not recognized as the pathological agent because of the presence of other more frequently involved bacteria and fungi . After multiple intravenous and oral antibiotics and antifungal treatments failed, further management involved frequent debridement of both external auditory canals and tympanic membranes, right tympanoplasty, bilateral mastoidectomy, revision tympanomastoidectomies and left modified radical mastoidectomy . Antistaphylococcal therapy including ceftazidime, vancomycin, teicoplanin, clindamycin and rifampicin was tried . Following the modified radical radical mastoidectomy, normalization of the status of his ears took approximately 2 months and has since remained stable to date . His left ear is deaf because of vancomycin administration, while magnetic resonance imaging and gallium scintigraphy have shown persistent inflammation of the skull base. Transplantation, 1999 Oct 27, 68(8), 1107 - 11 Outcome of transplantation of organs procured from bacteremic donors; Freeman RB et al.; BACKGROUND: Transplantation of organs from donors who are bacteremic is controversial . We examined the outcome of recipients of solid organs from donors with bacteremia and/or fungemia at the time of organ recovery . METHODS: All organ donors from a single organ procurement organization between January 1990 and December 1996 were retrospectively analyzed . We calculated rates of transmission from bacteremic or fungemic donors to their recipients and compared the graft and patient survival rates for recipients of these organs with those for recipients of organs from non-bacteremic donors . RESULTS: There were 95 (5.1%) bacteremic donors from a total of 1775, from whom 212 recipients received organs . Forty-six (48%) of the bacteremic donors had pathogens in their blood . Among the 101 recipients of organs from these, no evidence of transmission could be documented . (0% transmission rate, 95% CI 0-3) . The remaining 49 donors had either Staphylococcus epidermidis or other unlikely pathogens recovered from the blood . Examination of the 111 recipients of organs from these donors also found no evidence for transmission (0% transmission rate, 95% CI 0-3) . Of the 212 recipients, 193 (91%) received a mean of 3.8+/-2.5 days of antibiotics postoperatively . The 30-day graft and patient survival for recipients of organs from bacteremic donors was not significantly different from recipients of organs from nonbacteremic donors (P = 0.695 for patient survival, and P = 0.310 for graft survival) . CONCLUSIONS: Organs transplanted from bacteremic donors do not transmit bacterial infection or result in poorer outcomes . Use of organs from these donors could help increase organ availability. Jpn J Cancer Res, 1999 Sep, 90(9), 928 - 33 Prevention of lung metastasis by intra-tumoral injection of Cepharanthin and staphylococcal enterotoxin B in transplantable rat osteosarcoma; Okada K et al.; The antitumor effect of intra-tumoral injection of Cepharanthin, a biscoclaurin alkaloid extracted from Stephania cephalanta Hayata, and staphylococcal enterotoxin B was evaluated using F344 male rats bearing transplantable rat osteosarcoma, S-SLM . A macroscopic lung metastatic nodule of tumor was transplanted into the subcutaneous back space, and 0.5 mg of Cepharanthin and 2 pg of staphylococcal enterotoxin B were injected into the tumor on days 12, 13 and 14 . On day 28, all animals were killed with an overdose of pentobarbital sodium, and the transplanted tumors and lungs were examined . The wet weight of the lungs of the rats treated with Cepharanthin and staphylococcal enterotoxin B was significantly lower, and apoptosis in the lung metastatic nodules was significantly higher than that of the control or that of rats treated with only Cepharanthin or staphylococcal enterotoxin B . In the transplanted tumors, infiltration of TRAP (tartrate-resistant acid phosphatase)-positive multinucleated giant cells was prominent in the rats treated with Cepharanthin and staphylococcal enterotoxin B . These findings indicate that intra-tumoral injection of Cepharanthin and staphylococcal enterotoxin B induced infiltration of TRAP-positive multinucleated giant cells within the transplanted rat osteosarcoma, and reduced lung metastasis. Int Endod J, 1999 Sep, 32(5), 370 - 5 Bacterial leakage versus dye leakage in obturated root canals; Barthel CR et al.; AIM: The aim of this study was to compare in vitro bacterial and dye leakage tests, commonly used to determine the seal of root canal fillings . METHODOLOGY: Ninety-six single-rooted teeth had their crowns removed at the cemento-enamel junction and their roots instrumented to ISO size 60 within 1 mm of the apex . Three groups of 30 roots were obturated by lateral condensation using gutta-percha and one or other of the sealers . AH26, Ketac Endo, or Roth's 801 sealer . Three roots were used as negative controls and three roots as positive controls . The roots were then exposed at their coronal end first for 38 days to soy broth containing Staphylococcus epidermidis and thereafter for 48 h to basic fuchsin . Bacterial leakage was recorded when the challenging organism could be recovered from the apical end . Dye leakage was checked by microscopy of transverse sections of the apical tip at the end of the experiment . RESULTS: For the bacterial experiment, there was no significant difference amongst the three sealer groups . The dye experiment showed significantly greater leakage in the AH26 compared to the Ketac Endo group . No correlation between the results of the two tests could be seen . CONCLUSIONS: The results of this study suggest that the molecular size of the penetrating agent may not be the relevant parameter when attempting to determine an appropriate test for the sealability of root canal fillings. J Pediatr Surg, 1999 Oct, 34(10), 1510 - 3 Complications of percutaneous insertion of Hickman catheters in children; Skladal D et al.; BACKGROUND/PURPOSE: The aim of this study was a retrospective evaluation of insertion and management complications of percutaneous Hickman catheter lines in pediatric patients to investigate whether the complication rate is acceptable in comparison with other insertion methods or other age groups . METHODS: Over a period of 22 months a total of 27 Hickman catheters were inserted in 22 pediatric patients (20 oncological, 2 nononcological; age 6 weeks to 17.5 years) . RESULTS: Twenty-three of 36 insertion attempts (63.9%) were successful at first attempt . In another 4 patients, catheters were placed after repeated attempts . In an additional 4 patients, catheters were inserted by surgeons after percutaneous insertion failed . As immediate complications, 1 pneumothorax and 1 malposition were seen . Late complications included 1 to 29 (median, 8) days of fever in 15 patients, corresponding to 53 of 1,000 catheter days . Fourteen patients showed 21 positive blood cultures, including 11 cases of Staphylococcus epidermides, which might be related to the catheter . Antibiotics were given for a total of 1 to 130 (median, 35) days, that is 205 of 1,000 catheter days . No catheter was removed because of infectious complications . The total life span of the Hickman catheters was 1 to 371 (median, 163) days, the patients were in the hospital from 1 to 351 (median, 102) days because of their underlying disease . At the end of the study period, 8 of 27 (29.6%) catheters remained functioning in situ; 9 (33.3%) had been selectively removed . Two patients died with the catheter (7.4%) functioning well . Another 2 patients showed catheter thrombosis . Six catheters (22.2%) in 5 patients showed inadvertent dislodgement . CONCLUSION: Percutaneous Hickman catheter insertion in pediatric patients is effective; however, complication rate is relevant, but not higher than percutaneous insertion of subclavian vein or Hickman catheters in adults. Protein Expr Purif, 1999 Nov, 17(2), 290 - 8 Tandem affinity tags for the purification of bivalent anti-DNA single-chain Fv expressed in Escherichia coli; Cocca BA et al.; Antibodies to DNA define an important autospecificity that arises in systemic lupus erythematosus (SLE) . To elucidate the molecular features that may explain the pathogenesis of SLE, a heterologous system for expression of cloned V genes is often desirable . Here, a single-chain Fv coding domain was constructed by using the heavy- and light-chain V genes of a high-affinity site-directed mutant of the murine anti-dsDNA autoantibody, 3H9 . This scFv was joined in frame to the c-jun leucine zipper for dimerization, and to two affinity tags, domain B of the staphylococcal protein A and a pentahistidine peptide, for purification . Dimerization of the scFv was determined by size-exclusion chromatography . The yields of the scFv following affinity purification on IgG agarose or Ni-NTA agarose were compared, and the activities of the resulting protein fractions were determined . A two-step purification of periplasmic extracts on Ni-NTA agarose and IgG agarose, followed by elution with 3.5 M MgCl(2), yielded scFv with the highest specific activity . The final purified material bound DNA by ELISA, electrophoretic mobility shift assay, and immunofluorescence of fixed Hep-2 cells . Antibodies purified in this fashion should have applications in structure/function studies in which it is essential to generate highly purified antigen-combining sites . Cutis, 1999 Oct, 64(4), 250 - 2 Leukocytoclastic vasculitis following staphylococcal protein A column immunoadsorption therapy for idiopathic thrombocytopenic purpura; Bourelly PE et al.; A patient with refractory idiopathic thrombocytopenic purpura developed widespread palpable purpura, fever, diarrhea, and arthralgias forty-eight hours following a third protein A immunoadsorption treatment . A skin biopsy revealed leukocytoclastic vasculitis . His skin lesions and constitutional symptoms resolved with administration of antibiotics and bed rest. Eur J Immunol, 1999 Oct, 29(10), 3262 - 72 Homeostatic regulation of CD8+ T cells by perforin; Kagi D et al.; To prevent uncontrolled expansion, the massive proliferation of T cells during an acute immune response has to be followed by controlled deletion . Here we show that similar to Fas, perforin is not only an important effector molecule of cytotoxic T lymphocytes (CTL) but also involved in down-regulating peripheral T cells . Mice deficient for both the CTL effector molecule perforin and the apoptosis-inducing Fas ligand spontaneously develop infiltration of highly activated CD8(+) T cells in kidney and liver and die between 5 and 12 weeks of age . Injection of staphylococcal enterotoxin B (SEB) into perforin-deficient mice results in dramatically increased selective expansion and prolonged persistence of CD8(+), but not CD4(+), SEB-reactive T cells . Also, secondary immunization of TCR transgenic perforin-deficient mice with the lymphocytic choriomeningitis virus glycoprotein-derived epitope peptide leads to an increased proliferation of transgenic CD8(+) T cells, that is not explained by failure to deplete professional antigen-presenting cells . These results point to a novel mechanism of T cell homeostasis in which the acquisition of perforin-dependent cytotoxic activity regulates the expansion and persistence of CD8(+) effector T cells in vivo. Microbiology, 1999 Oct, 145 ( Pt 10), 2881 - 9 Staphylococcal phosphoenolpyruvate-dependent phosphotransferase system--two highly similar glucose permeases in Staphylococcus carnosus with different glucoside specificity: protein engineering in vivo? Christiansen I, Hengstenberg W. Previous sequence analysis of the glucose-specific PTS gene locus from Staphylococcus carnosus revealed the unexpected finding of two adjacent, highly similar ORFs, glcA and glcB, each encoding a glucose-specific membrane permease EIICBA(Glc) . glcA and glcB show 73% identity at the nucleotide level and glcB is located 131 bp downstream from glcA . Each of the genes is flanked by putative regulatory elements such as a termination stem-loop, promoter and ribosome-binding site, suggesting independent regulation . The finding of putative cis-active operator sequences, CRE (catabolite-responsive elements) suggests additional regulation by carbon catabolite repression . As described previously by the authors, both genes can be expressed in Escherichia coli under control of their own promoters . Two putative promoters are located upstream of glcA, and both were found to initiate transcription in E . coli . Although the two permeases EIICBA(Glc)1 and EIICBA(Glc)2 show 69% identity at the protein level, and despite the common primary substrate glucose, they have different specificities towards glucosides as substrate . EIICBA(Glc)1 phosphorylates glucose in a PEP-dependent reaction with a Km of 12 microM; the reaction can be inhibited by 2-deoxyglucose and methyl beta-D-glucoside . EIICBA(Glc)2 phosphorylates glucose with a Km of 19 microM and this reaction is inhibited by methyl alpha-D-glucoside, methyl beta-D-glucoside, p-nitrophenyl alpha-D-glucoside, o-nitrophenyl beta-D-glucoside and salicin, but unlike other glucose permeases, including EIICBA(Glc)1, not by 2-deoxyglucose . Natural mono- or disaccharides, such as mannose or N-acetylglucosamine, that are transported by other glucose transporters are not phosphorylated by either EIICBA(Glc)1 nor EIICBA(Glc)2, indicating a high specificity for glucose . Together, these findings support the suggestion of evolutionary development of different members of a protein family, by gene duplication and subsequent differentiation . C-terminal fusion of a histidine hexapeptide to both gene products did not affect the activity of the enzymes and allowed their purification by Ni2+-NTA affinity chromatography after expression in a ptsG (EIICB(Glc)) deletion mutant of E . coli . Upstream of glcA, the 3' end of a further ORF encoding 138 amino acid residues of a putative antiterminator of the BglG family was found, as well as a putative target DNA sequence (RAT), which indicates a further regulation by glucose specific antitermination. Int J Oncol, 1999 Nov, 15(5), 873 - 82 Treatment with tumor-reactive Fab-IL-2 and Fab-staphylococcal enterotoxin A fusion proteins leads to sustained T cell activation, and long-term survival of mice with established tumors; Sogaard M et al.; C215Fab-IL-2 fusion protein, with full IL-2 and antigen binding activity, was produced in E . coli at high level (>50 mg/l) . When co-administered with Fab-superantigen fusion protein (C215Fab-SEA) in mice strong and sustained T cell activation was observed . Combination treatment of mice carrying B16 melanoma transfected with C215 antigen was also more efficient than using C215Fab-SEA (p<0.01) or C215Fab-IL-2 alone (p<0.001) . In a long-term survival experiment 5/12 mice having received combination treatment 5 days after i.v . inoculation of B16 cells survived >85 days . Improved therapeutic efficacy correlated with increased tumor infiltration by activated CD25+ T cells, indicating a T cell mediated mechanism. Hepatology, 1999 Nov, 30(5), 1241 - 51 Concanavalin A hepatotoxicity in mice: tumor necrosis factor-mediated organ failure independent of caspase-3-like protease activation; Kunstle G et al.; Several models of tumor necrosis factor (TNF)/TNF-receptor 1 (TNF-R1)-dependent liver injury in mice were investigated with respect to caspase-3-like protease activation representing a pivotal mechanism of apoptotic cell death . Injection of TNF or T-cell-activating agents (i.e., agonistic anti-CD3 antibody or staphylococcal enterotoxin B {SEB}) into galactosamine (GalN)-sensitized mice caused TNF/TNF-R1-dependent liver injury . Intravenous concanavalin A (Con A) alone induced TNF-mediated hepatotoxicity dependent on both TNF-R1 and TNF-R2 . Hepatic caspase-3-like proteases were activated in GalN/TNF, GalN/anti-CD3, or GalN/SEB-treated mice, but not in Con A-treated mice . Consistently, the broad-spectrum caspase inhibitor, benzoyloxycarbonyl-val-ala-asp-fluoromethylketone (zVADfmk), prevented TNF-mediated hepatotoxicity in all GalN-dependent models, but failed to protect against Con A . Under transcriptional arrest, however, Con A induced TNF-R1-dependent, but not TNF-R2-dependent, activation of caspase-3-like proteases, and zVADfmk prevented animals from Con A-mediated liver injury under this condition . Histological analysis revealed distinct differences between Con A- and GalN/Con A-induced liver injury regarding apoptotic morphology of hepatocytes . We conclude that impaired transcription induces a switch of Con A hepatotoxicity toward a caspase-3-like protease-dependent pathway . The observation that the functional state of the transcriptional machinery decides whether TNF-driven hepatocyte apoptosis involves activation of caspase-3-like proteases or alternative signaling pathways in vivo might be of relevance for the immunopathology of the liver. Transplantation, 1999 Oct 15, 68(7), 1018 - 23 FK506 markedly enhances apoptosis of antigen-stimulated peripheral T cells by down-regulation of Bcl-xL; Migita K et al.; BACKGROUND: FK506 is a clinically effective immunosuppressive agent and promoter of immunologic tolerance . However, limited information is available about the mechanism of FK506-induced immunosuppression . METHODS: In the present study, we investigated the molecular mechanism of FK506-mediated enhancement of apoptosis using in vivo activated T lymphocytes . We examined the effects of FK506 on apoptosis-related proteins in superantigen-stimulated peripheral T cells . Results . Injection of staphylococcal enterotoxin B (SEB) into BALB/c mice resulted in a selective apoptosis of splenic Vbeta8-positive T cells after 48 hr . Injection of FK506 within 36 hr of SEB injection resulted in a marked enhancement of DNA fragmentation of splenic Vbeta8+ T cells . FK506 did not affect the expression of Fas antigen on SEB-activated Vbeta8+ T cells . As Bcl-2-related proteins are involved in apoptotic process, we also evaluated their role by examining the expression of Bcl-2, Bcl-X(L), and Bax on SEB-FK506-treated murine splenic T cells . Although SEB injection slightly increased the expressions of Bcl-2 and Bax on V138+ T cells, FK506 did not modulate Bcl-2 or Bax expression in these cells . In contrast, the expression of Bcl-x(L) on Vgamma8+ T cells, which was markedly induced by SEB, was abrogated by FK506 . Conclusions . Our findings indicate FK506-induced enhancement of apoptosis of activated T cells is mediated by down-regulation of Bcl-X(L) expression on these cells . Our results also suggest that Bcl-x(L) is a critical determinant of apoptosis of activated T cell and may represent a potential target for new therapies designed to achieve immunological tolerance. J Bone Joint Surg Br, 1999 Sep, 81(5), 886 - 9 Intraoperative bacterial contamination in operations for joint replacement; Davis N et al.; All surgical operations have the potential for contamination, and the equipment used can harbour bacteria . We collected samples from 100 elective primary hip and knee arthroplasties . These showed rates of contamination of 11.4% for the sucker tips, 14.5% for light handles, 9.4% for skin blades and 3.2% for the inside blades used during surgery; 28.7% of gloves used for preparation were also contaminated . Of the samples taken from the collection bags used during hip arthroplasty, 20% grew bacteria, which represents a significant microbial reservoir . Also, 17% of theatre gowns were contaminated at the end of the operation . Contamination was found in 10% of the needles used during closure of the fascia . Overall, 76% of the organisms grown were coagulase-negative staphylococcus . A total of 63% of operations showed contamination in the field of operation . Some changes in practice are suggested . Follow-up for a minimum of two years revealed one deep infection but the organism was not identified as a contaminant . These data provide a baseline for studying the bacteriology of the surgery of revision arthroplasty. Vox Sang, 1999, 77 Suppl 1, 57 - 64 Extracorporeal immunoadsorption for the treatment of haemophilic patients with inhibitors to factor VIII or IX; Knobl P et al.; BACKGROUND AND OBJECTIVES: This article reviews the relevance of immunoadsorption in the treatment of haemophilic patients with inhibitors . MATERIALS AND METHODS: Immunosorba (sepharose-bound staphylococcal protein A) and Ig-Therasorb (sepharose-bound polyclonal sheep antibodies to human immunoglobulin) columns are suitable for the clinical use of immunoadsorption . They allow the processing of large plasma volumes (>7,000 ml) without relevant side-effects . RESULTS: A haemophilic patient was treated with the Malmo protocol, another was admitted with intracerebral bleeding . Immunoadsorption reduced the inhibitor titer by 70-90% . CONCLUSIONS: Immunoadsorption can be used in cases of acute bleeding, before surgery, acquired factor VIII (FVIII) antibodies, and before the start of immune tolerance therapy . We suggest the inclusion of this method in immune tolerance protocols in order to improve levels, recovery, and half-life of FVIII and to save concentrates. Eur J Pharmacol, 1999 Sep 17, 381(1), 77 - 84 Structural determinants of phorbol ester binding in synaptosomes: pharmacokinetics and pharmacodynamics; Murphy TV et al.; The present study used structurally distinct phorbol esters to investigate the relationship between their pharmacokinetics of binding to protein kinase C (PKC) in rat brain cortex synaptosomes, their affinity for PKC in synaptosomes and ability to enhance noradrenaline release from rat brain cortex . Affinity binding studies using {3deoxyphorbol 13-tetradecanoate (dPT)=PDB&z . Gt;12-deoxyphorbol 13-acetate (dPA)=phorbol 12,13-diacetate (PDA) . In intact synaptosomes PDB, dPA and PDA rapidly displaced bound {3H}PDB whereas PMA and dPT were comparatively slow . However, the displacement rates for all the phorbol esters were equally rapid in synaptosomal membranes or synaptosomes permeabilised with Staphylococcus alpha-toxin . These results suggest that the lipophilic phorbol esters (dPT and PMA) are slower to displace {3H}PDB binding because they are hindered by the plasma membrane . In brain cortex slices it was found that the rate of displacement of {3H}PDB binding was closely correlated with the degree of elevation of transmitter noradrenaline release . Thus kinetic characteristics may determine biological responses and this may be particularly evident in events which occur rapidly or where there is fast counter-regulation. J Ren Nutr, 1999 Oct, 9(4), 206 - 13 Successful intradialytic parenteral nutrition after abdominal "Catastrophes" in chronically hemodialysed patients; Korzets A et al.; OBJECTIVE: To assess the therapeutic contribution of intradialytic parenteral nutrition (IDPN) in four acutely ill, hypercatabolic, hemodialysed patients . All underwent major surgery, complicated by infection and malnutrition . DESIGN: A retrospective clinical study . SETTING: An in-center hemodialysis unit, at a tertiary referral hospital . PATIENTS: Patient 1: a young woman, with a good renal transplant . Developed gastric lymphoma, which required gastrectomy . After cessation of immunosuppression, "lost" her kidney and returned to hemodialysis . Received IDPN for 4 months and recovered well from severe malnourishment . Patient 2: an elderly, malnourished man, on continuous ambulatory peritoneal dialysis (CAPD) . Developed biliary peritonitis and bacteremia . In a 3-month period, the patient had four operations . Maintained on IDPN for 4 months . Patient 3: a young and obese man, who suffered from life-threatening staphylococcal aureus peritonitis, resulting in widespread bowel adhesions . Underwent repeated aspirations of purulent ascites, laparoscopy, and explorative laparotomy . IDPN was administered for 4 months and stopped on the patient's request . Patient 4: a young man, who after cadaveric renal transplantation remained hospitalized for 6 months because of acute rejection and peritoneal and retroperitoneal abscesses . Had major surgery performed seven times . Received IDPN for 6 months, and is now well . RESULTS: All four patients benefited from 4 to 6 months of IDPN, as an integral part of intensive supportive and nutritional treatment . Weight loss was halted, as patient appetite returned and oral nutrition became adequate . Estimated daily protein intake reached 1.2 g/kg, while caloric intake rose to nearly 30 kcal/kg/d (Table 3) . Mean serum albumin levels increased from 25.5 g/L +/- 0.9 g/L to 38.0 g/L +/- 1.5 g/L . No adverse side effects were seen from IDPN . CONCLUSION: IDPN is a worthwhile part of treatments used in the catabolic, postoperative hemodialysed patient . It is safe and efficient when used over a 6-month period in trying to attenuate existing, or worsening malnutrition in these patients . It should be commenced at an early stage in these patients, after attempts at oral nutritional support have been deemed inadequate. J Neurol, 1999 Sep, 246(9), 815 - 20 Spinal epidural abscess complicating chronic epidural analgesia in 11 cancer patients: clinical findings and magnetic resonance imaging; Sillevis Smitt P et al.; We reviewed the records of all patients who had received an epidural catheter for management of chronic cancer pain in a 3-year period (1993-1996) . Patients with nervous system infections were identified, and pertinent clinical, radiological (magnetic resonance imaging), and bacteriological data were analyzed . We identified 11 patients who developed spinal epidural abscess (SEA) . All of these had back pain; radicular signs occurred in seven patients and spinal cord compression in two patients . Magnetic resonance imaging revealed SEA in all 11 patients . SEA was iso- to hypointense on T1-weighted images and hyperintense on T2-weighted images relative to spinal cord . After gadolinium administration seven lesions showed characteristic rim enhancement while three showed minimal enhancement . No signs of diskitis or osteomyelitis were present, and the abscess was always localized to the posterior epidural space . Cultures were positive in all cases and revealed Staphylococcus epidermidis in eight and S . aureus in three . All patients were treated with intravenous antibiotics, and four had an additional decompressive laminectomy . Two patients died within 1 week of diagnosis from overwhelming septicemia despite apparently adequate antibiotic treatment . Within 4 weeks after diagnosis of SEA two patients died from widely metastatic disease, although infection may have contributed . One patient developed septicemia while receiving appropriate antibiotics and underwent emergency laminectomy . The neurological deficits recovered in all patients who survived the acute infectious episode . We conclude that patients with chronic epidural catheters for cancer pain require prompt neurological evaluation and magnetic resonance imaging when SEA is suspected . Early evaluation and treatment may lead to full recovery. J Pharmacol Exp Ther, 1999 Nov, 291(2), 680 - 7 RWJ 67657, a potent, orally active inhibitor of p38 mitogen-activated protein kinase; Wadsworth SA et al.; Tumor necrosis factor-alpha (TNF-alpha), a cytokine secreted by activated monocytes/macrophages and T lymphocytes, has been implicated in several disease states, including rheumatoid arthritis, inflammatory bowel disease, septic shock, and osteoporosis . Monocyte/macrophage production of TNF-alpha is dependent on the mitogen-activated protein kinase p38 . RWJ 67657 (4-{4-(4-fluorophenyl)-1-(3-phenylpropyl)-5-(4-pyridinyl)-1H-imidazol -2-yl}-3-butyn-1-ol) inhibited the release of TNF-alpha by lipopolysaccharide (a monocyte stimulus)-treated human peripheral blood mononuclear cells with an IC(50) of 3 nM, as well as the release of TNF-alpha from peripheral blood mononuclear cells treated with the superantigen staphylococcal enterotoxin B (a T cell stimulus), with an IC(50) value of 13 nM . This compound was approximately 10-fold more potent than the literature standard p38 kinase inhibitor SB 203580 in all p38 dependent in vitro systems tested