J Microbiol Methods, 2003 Jul, 54(1), 105 - 10 Comparative studies on the activity of basil--an essential oil from Ocimum basilicum L.--against multidrug resistant clinical isolates of the genera Staphylococcus, Enterococcus and Pseudomonas by using different test methods; Opalchenova G et al.; The essential oil basil is obtained from the aerial parts of Ocimum basilicum L . After gas chromatographic separation, the following components were identified: linalol (54.95%), methylchavikol (11.98%), methylcinnamat (7.24%) and linolen (0.14%) . The activity of basil against multidrug resistant clinical isolates from the genera Staphylococcus, Enterococcus and Pseudomonas has been studied . For this purpose, standard and modified broth macrodilution methods were used and time kill kinetic of basil was studied . The minimum inhibitory concentrations (MICs) were reported between 0.0030% and 0.0007% (v/v) . These concentrations were compared with the inhibitory concentrations (ICs) and the logs of the bacterial counts reduction both obtained by basil diluted in 1% Tween (Tw) 80, saline test solution (STS) and spiritus vini (Sv) 95 degrees instead in a broth . The data, obtained after application of different methods of investigation and validated with membrane filtration, showed a strong inhibitory effect of basil on the test bacteria . The chosen bacteria are widespread and pose serious therapeutic difficulties because of their high level of resistance . For this reason, the results obtained were considered encouraging. Biochemistry, 2003 May 13, 42(18), 5395 - 402 Binding of nonphysiological protein and peptide substrates to proteases: differences between urokinase-type plasminogen activator and trypsin and contributions to the evolution of regulated proteolysis; Bergstrom RC et al.; Understanding the regulation of physiological processes requires detailed knowledge of the recognition of substrates by enzymes . One of the most productive model systems for the study of enzyme-substrate interactions is the serine protease family; however, most studies of protease action have used small substrates that contain an activated, non-natural scissile bond . Because few kinetic or structural studies have used protein substrates, the physiologically relevant target of most proteases, it seems likely that important mechanisms of substrate recognition and processing by proteases have not yet been fully elucidated . Consistent with this hypothesis, we have observed that K(m) values for protein substrates are reduced as much as 200-15000-fold relative to those of analogous peptide substrates . Here we examine the thermodynamic consequences of interactions between proteases and their substrates using staphylococcal nuclease (SNase) and SNase variants as model protein substrates . We have obtained values for enthalpy, entropy, and K(d) for binding of proteins and peptides by the nonspecific protease trypsin and the highly specific protease urokinase-type plasminogen activator (u-PA) . To avoid cleavage of substrates during these measurements, we used inactive variants of trypsin and u-PA whose catalytic serine S195 had been replaced by alanine . Differences in the K(d) values for binding of protein and peptide substrates closely approximate the large differences observed in the corresponding K(m) values . Improved binding of protein substrates is due to decreased enthalpy, and this effect is pronounced for the selective protease u-PA . Fundamental differences in recognition of analogous protein and peptide substrates may have influenced the evolution of protease specificity. Anal Biochem, 2003 Jun 1, 317(1), 107 - 15 General properties of GFP-display, an electrophoretic analysis for single amino acid changes in target polypeptides; Aoki T et al.; The migrating position of green fluorescent protein (GFP)-fused polypeptide varied on an SDS/urea gel by a single amino acid change in the fused polypeptide segment . An easy detection method for a single amino acid change based on this observation was called "GFP-display." Using various target polypeptides, staphylococcal protein A (SpA), Ras, p53, and human beta3 adrenergic receptor (AR), and their mobility-shift patterns resulting from the single amino acid changes, several important properties of GFP-display were revealed as follows: (i) . since the binding of dodecyl sulfate ions to acidic or hydrophilic amino acids is weaker than that to basic or hydrophobic amino acids, the ions bound weakly to the fused polypeptide segment are forced to come off by high concentrations of urea prior to the ions bound strongly, resulting in the mobility shift, (ii) . the mobility shift is estimated to a certain extent using a new parameter called the "GD value" calculated from the isoelectric point, hydrophilicity, and number of fused amino acids, and (iii) . the fluorescence intensity of GFP-fused polypeptide tends to increase with the average hydrophilicity of the fused polypeptide segment . GFP-display will be a helpful technique for many kinds of gene or protein studies related to amino acid substitutions such as the random mutagenesis in a gene of interest. Chemphyschem, 2003 Apr 14, 4(4), 359 - 65 On the temperature--pressure free-energy landscape of proteins; Ravindra R et al.; We studied the thermodynamic stability of a small monomeric protein, staphylococcal nuclease (Snase), as a function of both temperature and pressure, and expressed it as a 3D free-energy surface on the p,T-plane using a second-order Taylor expansion of the Gibbs free-energy change delta G upon unfolding . We took advantage of a series of different techniques (small-angle X-ray scattering, Fourier-transform infrared spectroscopy, differential thermal analysis, pressure perturbation calorimetry and densitometry) in the evaluation of the conformation of the protein and in evaluating the changes in the thermodynamic parameters upon unfolding, such as the heat capacity, enthalpy, entropy, volume, isothermal compressibility and expansivity . The calculated results of the free-energy landscape of the protein are in good agreement with experimental data of the p,T-stability diagram of the protein over a temperature range from 200 to 400 K and at pressures from ambient pressure to 4000 bar . The results demonstrate that combined temperature--pressure-dependent studies can help delineate the free-energy landscape of proteins and hence help elucidate which features and thermodynamic parameters are essential in determining the stability of the native conformational state of proteins . The approach presented may also be used for studying other systems with so-called re-entrant or Tamman loop-shaped phase diagrams. Vet Pathol, 2003 May, 40(3), 283 - 7 Abscess-forming inflammatory granulation tissue with Gram-positive cocci and prominent eosinophil infiltration in cats: possible infection of methicillin-resistant Staphylococcus; Ozaki K et al.; We occasionally encounter feline cervical or mesenteric lesions diagnosed histopathologically as abscess or inflammatory granulation tissue with eosinophil infiltration . Gram-positive cocci accompany the lesions . In the present study, such lesions obtained from 27 cats were examined to evaluate the histopathologic features and the nature of the causative bacteria . The average age was 7.3 +/- 3.5 years . No sex predilection was observed . Most frequent locations of the lesions included the abdominal cavity with/without mesenteric lymph nodes (11/27, 41%) and subcutaneous tissue or lymph nodes of the neck (9/27, 33%) . Common clinical presentation was a localized mass . Grossly, the lesions contained abscesses in the center and were surrounded by fibrous tissue . Microscopically, the necrotic zone contained bacterial colonies . Large numbers of eosinophils and macrophages infiltrated the area surrounding the necrotic tissue . The surrounding connective fiber-rich granulation tissue demarcated the eosinophilic abscess . The bacteria were Gram-positive cocci in 23 of the 27 cats and were positive for anti-staphylococcus antiserum in 19 of the 23 cats . In 15 out of 17 lesions, the colonies expressed immunoreactivity to penicillin-binding protein 2', which is a drug-resistance gene product of methicillin-resistant Staphylococcus (MRS) species . These findings suggest strongly that MRS causes this type of infectious lesion. Clin Infect Dis, 2003 May 1, 36(9), 1157 - 61 Epub 2003 Apr 14. Prosthetic joint infections: bane of orthopedists, challenge for infectious disease specialists; Lentino JR; Prosthetic joint infections (PJIs) occur in approximately 1.5%-2.5% of all primary hip or knee arthroplasties . The mortality rate attributed to PJIs may be as high as 2.5% . Substantial morbidity is associated with a loss of mobility, although this is temporary . The costs associated with a single episode of PJI are approximately $50,000 per episode, exclusive of lost wages . Risk factors that increase the occurrence of PJI include revision arthroplasty, time in the operating room, postoperative surgical site infection, and malignancy . Pain is the most consistent symptom . Staphylococcus species are the most common organisms isolated from PJI sites . Two-stage revision is superior to single-stage revision or to debridement with prosthesis retention . Long-term antibiotic suppression and/or arthrodesis are useful for patients too frail to undergo extensive surgery . Using an optimal approach, recurrent infection occurs in <10% of previously infected joints. Hybrid Hybridomics, 2003 Feb, 22(1), 33 - 9 Generation and characterization of a mouse monoclonal antibody with specificity similar to staphylococcal protein A (SPA); Hadji-Ghasemi F et al.; Human IgG is comprised of four subclasses (IgG(1), IgG(2), IgG(3), and IgG(4)) . Each subclass possesses different biological properties . One of the differential specificities of human IgG subclasses is binding of Fc fragment of IgG(1), 2, and 4 but, not IgG(3) to staphylococcal protein A (SPA) . This study was conducted to produce, select and characterize a monoclonal antibody (MAb) recognizing human IgG subclasses with specificity similar to SPA . Splenocytes from Balb/c mice immunized with Fc fraction of a human IgG(1) myeloma protein were fused with Sp2/0 myeloma cells . Fused cells were grown in hypoxanthine, aminopterine, and thymidine (HAT) selective medium and cloned by limiting dilution assay . Antibody-secreting cells were screened by enzyme-linked immunosorbent assay (ELISA) and the specificity of secreted MAb was further analyzed, using a panel of purified myeloma proteins by ELISA and immunoblotting . A murine hybridoma designated 6F11E1 was obtained that secretes an MAb specific for the Fc fragment of the immunizing protein . This MAb reacts with isotypic epitope common to IgG(1), 2 and 4 subclasses . An allelic epitope linked to IgG(3) molecules is also recognized by 6F11E1 . This pattern of reactivity was found to be highly similar to that of SPA . Our findings imply that similar or overlapping epitopes are recognized by 6F11E1 and SPA. J Biomater Sci Polym Ed, 2003, 14(3), 279 - 95 Inhibition of bacterial and leukocyte adhesion under shear stress conditions by material surface chemistry; Patel JD et al.; Biomaterial-centered infections, initiated by bacterial adhesion, persist due to a compromised host immune response . Altering implant materials with surface modifying endgroups (SMEs) may enhance their biocompatibility by reducing bacterial and inflammatory cell adhesion . A rotating disc model, which generates shear stress within physiological ranges, was used to characterize adhesion of leukocytes and Staphylococcus epidermidis on polycarbonate-urethanes and polyetherurethanes modified with SMEs (polyethylene oxide, fluorocarbon and dimethylsiloxane) under dynamic flow conditions . Bacterial adhesion in the absence of serum was found to be mediated by shear stress and surface chemistry, with reduced adhesion exhibited on materials modified with polydimethylsiloxane and polyethylene oxide SMEs . In contrast, bacterial adhesion was enhanced on materials modified with fluorocarbon SMEs . In the presence of serum, bacterial adhesion was primarily neither material nor shear dependent . However, bacterial adhesion in serum was significantly reduced to < or = 10% compared to adhesion in serum-free media . Leukocyte adhesion in serum exhibited a shear dependency with increased adhesion occurring in regions exposed to lower shear-stress levels of < or = 7 dyne/cm2 . Additionally, polydimethylsiloxane and polyethylene oxide SMEs reduced leukocyte adhesion on polyether-urethanes . In conclusion, these results suggest that surface chemistry and shear stress can mediate bacterial and cellular adhesion . Furthermore, materials modified with polyethylene oxide SMEs are capable of inhibiting bacterial adhesion, consequently minimizing the probability of biomaterial-centered infections. Zh Mikrobiol Epidemiol Immunobiol, 2000 Jul-Aug, (4 Suppl), 59 - 62 {Effect of carnosine on the morphofunctional state of mucosal cells of the soft palate of rats in staphylococcal infection}; Stadnikov AA et al.; The influence of carnosine on the morphofunctional state of mucosal cells of the soft palate of rats in experimental staphylococcal infection has been studied . Morphological changes in muscular and connective tissue cells of the mucosa of the soft palate of rats have been found to occur . The suppressive effect of carnosine with respect to the pathogen and its positive influence on the processes of the regeneration of eukaryotic tissues. Zh Mikrobiol Epidemiol Immunobiol, 2000 Jul-Aug, (4 Suppl), 56 - 9 {Biological importance of the anti-carnosine activity of bacteria}; Bukharin OV et al.; The anticarnosine activity has been found to be widely spread among bacteria, the degree of its manifestation depending on the source of isolation . The fact that anticarnosine activity plays a certain role in the phenomenon of persistence has been proved on the model of experimental staphylococcal infection in rats. Immunology, 2003 May, 109(1), 24 - 31 Telomere length measurement and determination of immunosenescence-related markers (CD28, CD45RO, CD45RA, interferon-gamma and interleukin-4) in skin-homing T cells expressing the cutaneous lymphocyte antigen: indication of a non-ageing T-cell subset; Neuber K et al.; The purpose of this study was to investigate the immunosenescence of skin-homing T cells expressing the cutaneous lymphocyte antigen (CLA) . Peripheral blood lymphocytes from 72 healthy individuals (33 male and 39 female; median age 54 years; age-range: 18-94 years) were investigated . The expression of CD28, CD45RA and CD45RO, as well as intracellular interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) formation of CLA+ 'skin homing' T cells, was analysed . In addition, T cells were detected immunohistologically in skin specimens from 15 young and 15 old, healthy individuals . The relative telomere length (RTL) was measured by fluorescence in situ hybridization using flow cytometry (flow FISH) . The total number of CLA+ T cells was found to remain constant with increasing age . In contrast to peripheral blood T cells (CD3+ CLA-), which showed significantly decreased CD28 and CD45RA expression in donors > 60 years of age, no age-related alterations of either CD28+ CLA+ T cells or CD45RA+ CLA+ T cells were observed . In the group of donors > 60 years of age, the proportion of intracellular IFN-gamma-producing CD3+ CLA- cells showed a significant increase, whereas the number of IFN-gamma- and IL-4-producing CLA+ T cells was not affected by age . After stimulation with phytohaemagglutinin (PHA) or staphylococcal enterotoxin B (SEB), CLA+ T cells from old donors did not show a reduced response compared with CLA+ T cells from young donors . Additionally, the counts of T cells in healthy skin from young and old adults were statistically not different . Furthermore, the RTL was significantly shortened in enriched CD45RO+ CLA- T cells from healthy old individuals, but not in aged CLA+ T cells . The present data suggest that CLA+ T cells might be a T-cell subpopulation which does not undergo immunosenescence . This may explain why the intensity of inflammatory skin reactions (e.g . psoriasis or eczema) seems to be independent of the patients' age. Allergy, 2003 Apr, 58(4), 329 - 36 Selective insufficiency of IFN-gamma secretion in patients with hyper-IgE syndrome; Ito R et al.; BACKGROUND: Hyper-immunoglobulin E (IgE) syndrome is a complex immune deficiency characterized by chronic eczematous dermatitis, recurrent staphylococcal infections, pneumatoceles, reduced neutrophil chemotaxis, and variably impaired T cell function . Although decreased interferon-gamma (IFN-gamma) production in patients with hyper-IgE syndrome is pointed out and known as a cause of reduced neutrophil chemotaxis, precise mechanism of their inadequate production of IFN-gamma remains unknown . To elucidate the pathogenesis of the defective production of IFN-gamma in patients with hyper-IgE syndrome, we assessed the in vitro production and secretion of IFN-gamma by peripheral blood mononuclear cells (PBMCs) from patients with hyper-IgE syndrome . METHODS: Chemotaxis of neutrophils, mRNA levels of several cytokines, intracellular production and extracellular secretion of IFN-gamma, interleukin-2 (IL-2), and IL-4 by PBMCs from three patients with hyper-IgE syndrome were determined . RESULTS: The transcription of IFN-gamma mRNA and the production of its protein molecules progressed normally . However, selective insufficiency in the secretion of IFN-gamma molecules was found in patients with hyper-IgE syndrome.Confocal laser scanning microscopy clearly demonstrated the accumulation of IFN-gamma in patients with hyper-IgE syndrome . CONCLUSION: We demonstrated that there was a selective insufficiency in the secretion of IFN-gamma in patients with hyper-IgE syndrome . We hope that this fact would offer a new paradigm for understanding this disease. Otolaryngol Head Neck Surg, 2003 Apr, 128(4), 447 - 51 Multiple-use atomizers in outpatient otolaryngology clinics are not necessarily an infectious risk; Visosky AM et al.; OBJECTIVE: We sought to determine the risk of bacterial transmission from multiple-use atomizers in an outpatient otolaryngology clinic . STUDY DESIGN AND SETTING: Atomizers in 6 examination rooms were analyzed for bacterial contamination . Sterility of associated tubing from 1 atomizer was assessed . Multidose drug solutions in the atomizers (phenylephrine and tetracaine) were cultured multiple times over a 2-week period . Serial dilutions of the original sample were cultured to minimize inhibitory effects of antiseptic agents in the drug formulations . RESULTS: No bacteria were recovered from tubing cultures . Only 2 (0.6%) of 336 cultures of the drug solutions yielded bacterial growth . In both cases, coagulase-negative Staphylococcus was recovered in 1 of the dilutions prepared from original samplings . CONCLUSION: Multiple-use atomizers that contain phenylephrine and tetracaine solutions with bacteriostatic preservatives in an outpatient otolaryngology clinic do not yield significant bacterial growth and, when used as described, do not pose an infectious risk of bacterial transmission between patients. Infect Immun, 2003 May, 71(5), 2542 - 7 Staphylococcal enterotoxin B-induced acute inflammation is inhibited by dexamethasone: important role of CXC chemokines KC and macrophage inflammatory protein 2; Schramm R et al.; This study was conducted to examine the anti-inflammatory mechanisms of dexamethasone during leukocyte recruitment and expression of the CXC chemokines macrophage inflammatory protein 2 (MIP-2) (CXCL2) and cytokine-induced neutrophil chemoattractant (KC) (CXCL1) in staphylococcal enterotoxin B (SEB)-induced acute inflammation . To do this, SEB was injected into murine air pouches with or without dexamethasone pretreatment for 2 h . SEB induced infiltration of leukocytes in a dose- and time-dependent manner, with the maximal response observed after 4 h of treatment with 10 microg of SEB . The recruited leukocytes comprised more than 77% neutrophils . Moreover, SEB challenge (10 microg) provoked time-dependent secretion of CXC chemokines, which peaked after 1 h . Local administration of antibodies against MIP-2 and KC significantly reduced SEB-triggered neutrophil accumulation by 38 and 59%, respectively . Dexamethasone (10 mg kg(-1)) significantly decreased neutrophil recruitment by 82% and reduced secretion of MIP-2 and KC by 89 and 85%, respectively, in response to SEB challenge . Our data demonstrate that dexamethasone potently inhibits neutrophil recruitment in SEB-induced inflammation . Moreover, we provide evidence that MIP-2 and KC are key mediators in the neutrophil response to SEB . Furthermore, our findings demonstrate that dexamethasone attenuates SEB-induced expression of MIP-2 and KC . Thus, this study elucidates important signaling pathways of SEB-induced neutrophil recruitment and anti-inflammatory mechanisms of action of dexamethasone. Crit Care Med, 2003 Apr, 31(4), 1154 - 9 Chronic Staphylococcal enterotoxin B and lipopolysaccharide induce a bimodal pattern of hepatic dysfunction and injury; Beno DW et al.; OBJECTIVE: To determine the effect of chronic exposure to endotoxin (lipopolysaccharide) and Staphylococcal enterotoxin B on hepatic injury and function . DESIGN: Prospective, controlled trial . SETTING: Research laboratory in a university hospital . SUBJECTS: Male Sprague-Dawley rats weighing 325-350 g with chronic vascular and bile catheters . INTERVENTIONS: Chronically catheterized rats were treated daily with saline, 50 microg/kg Staphylococcal enterotoxin B alone, 1000 microg/kg lipopolysaccharide alone, 1000 microg/kg lipopolysaccharide with 50 microg/kg Staphylococcal enterotoxin B, or 100 microg/kg lipopolysaccharide with 50 microg/kg Staphylococcal enterotoxin B for 10 days . Serum and biliary measures of hepatic injury and dysfunction were measured before and then 6 hrs and 1, 2, 3, 7, and 10 days after the start of treatment . The animals were killed at 10 days and the livers examined histologically . MEASUREMENTS AND MAIN RESULTS: Mean rates of bile flow, biliary indocyanine green excretion, and bile acid flux were significantly decreased immediately after treatment (6 hr, 1 and 2 days) and then at 10 days . Increases in biliary and serum gamma-glutamyltransferase and serum bile acids also occurred in a similar bimodal pattern . Animals treated with lipopolysaccharide or Staphylococcal enterotoxin B alone became tolerant and did not develop the bimodal pattern of hepatic dysfunction . Histologic examination of the liver at 10 days revealed periportal inflammation and fibrosis . CONCLUSIONS: The combination of lipopolysaccharide and Staphylococcal enterotoxin B leads to late liver injury, whereas either toxin alone does not . These data may explain the frequent development of liver dysfunction in patients exposed to multiple bacterial toxins such as in sepsis, multiple-system organ failure, and other diseases with altered intestinal permeability. Cytometry A, 2003 May, 53(1), 28 - 38 Optimal preparation of rhesus macaque blood for cytokine flow cytometric analysis; Nomura LE et al.; BACKGROUND: The rhesus macaque is a common substitute for human subjects in many disease models, including simian immunodeficiency virus, the non-human primate equivalent of the human immunodeficiency virus . Monoclonal antibodies and fluorochromes optimized for use in macaques were included in samples examined for immune responses with the use of intracellular cytokine flow cytometry (CFC) . METHODS: Sample preparation was optimized based on the following comparisons: activation of peripheral blood mononuclear cells (PBMCs) versus whole blood; separation of PBMCs using BD Vacutainer cell preparation tubes versus Ficoll; and activation of samples on the day they were collected versus holding samples overnight . RESULTS: When activated with the simian immunodeficiency virus type mac239 and Gag peptide mix or with the superantigen Staphylococcal enterotoxin B, separated PBMCs produced greater CD4 and CD8 fluorescence intensities and a larger percentage of CD69+ cytokine-positive cells than did whole blood samples . PBMCs separated by cell preparation tubes produced absolute T-lymphocyte counts equivalent to that with Ficoll separation, and CFC results with both methods were similar . When subjected to overnight shipping conditions, whole blood and PBMCs sometimes showed a reduction in mean fluorescence intensity and percentage of CD69+ cytokine-positive T lymphocytes . CONCLUSIONS: Due to this reduction in responses, it is preferable to activate samples on the day of blood collection . Samples can be surface stained and frozen in BD FACS Lysing Solution, to be thawed at a later date; this preserves their ability to display a cytokine response . Thus optimal CFC results are achieved by separating macaque PBMCs from whole blood, activating samples on day of collection, and, if necessary, freezing samples after surface staining for future analysis . Pediatr Nephrol, 2003 Apr, 18(4), 378 - 83 Epub 2003 Mar 18. Chronic hemodialysis in infants and children under 2 years of age; Shroff R et al.; Over the past 16 years, 18 children under 2 years of age received chronic hemodialysis (HD) at our center . Five children were anuric at the start of HD and 6 had significant co-morbidity . The most common underlying diagnosis was posterior urethral valves . The median age at the start of HD was 12.2 months . A total of 39 episodes (defined as a discrete time period during which HD was the principle form of renal replacement therapy) of HD were performed, with a median duration of 7 months and 91.3 dialysis sessions per episode . Problems with vascular access were very common, with a revision ratio of 40% . Twenty-two line revisions were required for 36 episodes of line infection, with a median rate of line infection of 2.7 infections/patient years . The most commonly encountered organism was coagulase-negative Staphylococcus (69%) . Twenty-three lines needed revision due to poor line function, despite the routine use of heparin . The effectiveness of HD was assessed in 11 patients who received HD for a continuous period of 3 or more months . The median urea reduction rate was 72%, while the parathyroid hormone levels improved to within twice the upper limit of the reference range in 69% . While there was no significant change in the median weight and height standard deviation score (SDS), the median SDS for head circumference showed significant improvement ( P=0.04) . Both growth and developmental outcomes were strongly influenced by existing co-morbidity . Sixteen (89%) children were transplanted . Four (22%) children died, 3 after successful transplants . None of the deaths occurred on HD or resulted from its complications . In conclusion, HD in infants and small children is an effective and safe form of renal replacement therapy, but problems with vascular access limit its long-term use. Toxicology, 2003 May 3, 187(2-3), 229 - 38 Toxicity of the staphylococcal enterotoxin B mutants with histidine-to-tyrosine substitutions; Korolev S et al.; In this study we made a series of site-directed mutants of staphylococcal enterotoxin B (SEB), in which histidine residues in the molecule were replaced by tyrosine . The mutant genes were cloned and expressed, and the corresponding proteins were purified . These mutant proteins were tested for binding to human HLA-DR4 and for mitogenetic activity in mouse splenocyte culture . Toxicity of the proteins in vivo was evaluated in the actinomycin D-primed C3H/HeJ mouse model . We found that SEB mutant proteins with fewer than four histidine-to-tyrosine (his-to-tyr) substitutions retained toxic properties similar to wild-type SEB . However, studies showed that his-to-tyr substitution of four consecutive histidine residues eliminated SEB toxicity . Our results clearly show that this genetically modified SEB protein is non-toxic and justifies its further development as a component of a new, safer vaccine to prevent SEB intoxication. Surg Infect (Larchmt), 2002 Winter, 3(4), 359 - 65 Epidural catheter colonization is not associated with infection; Kostopanagiotou G et al.; BACKGROUND: Epidural anesthesia is one of the most common types of regional anesthesia . Although retrospective reviews suggest that the incidence of infection from short-term epidural catheter use is exceedingly low in patients undergoing surgery, the correlation between epidural catheter cultures and infection has not been well defined . The purpose of this study was to determine the frequency of bacterial colonization of epidural catheters in adult patients undergoing surgery under epidural anesthesia, as well as the correlation between epidural catheter cultures and infection . METHODS: A prospective nonrandomized study was conducted over a period of 28 months . The incidence of bacterial contamination after epidural catheterization and the correlation between epidural catheter cultures and infection was investigated for 245 adult ASA I, II patients undergoing surgery under lumbar epidural anesthesia . Catheters were removed when epidural analgesia was no longer required, or if clinical signs of infection or catheter malfunction were present . All epidural catheters were cultured upon withdrawal . RESULTS: Epidural catheters were kept in place for 2.3 +/- 0.2 days (range 0.1-12 days) . Bacteriological analysis of the 245 epidural catheters yielded 28% positive cultures . The most prevalent microorganism was Staphylococcus epidermidis (58%) . In obstetric operations a frequency of 32% positive cultures was observed . Neither central nervous system nor systemic infections occurred during the study . No correlations were found among the type of surgery, the type of antibiotic administration, the sex or age of the patients, the duration of catheter placement, the maximum body temperature, and the frequency of positive epidural catheter cultures . There was no correlation between epidural catheter colonization and infection . CONCLUSION: The contamination of epidural catheters was found to be independent of the administered antimicrobial agents prior to surgery, the duration of catheter placement, and the presence of fever . Except for perioperative prophylaxis, therapeutic use of antibiotics for short-term epidural catheters is not recommended. J Exp Med, 2003 Apr 21, 197(8), 1017 - 28 Epub 2003 Apr 14. Immune enhancement of skin carcinogenesis by CD4+ T cells; Daniel D et al.; In a transgenic model of multi-stage squamous carcinogenesis induced by human papillomavirus (HPV) oncogenes, infiltrating CD4+ T cells can be detected in both premalignant and malignant lesions . The lymph nodes that drain sites of epidermal neoplasia contain activated CD4+ T cells predominantly reactive toward Staphylococcal bacterial antigens . HPV16 mice deficient in CD4+ T cells were found to have delayed neoplastic progression and a lower incidence of tumors . This delay in carcinogenesis is marked by decreased infiltration of neutrophils, and reduced activity of matrix metalloproteinase-9, an important cofactor for tumor progression in this model . The data reveal an unexpected capability of CD4 T cells, whereby, proinflammatory CD4+ T cells, apparently responding to bacterial infection of dysplastic skin lesions, can inadvertently enhance neoplastic progression to invasive cancer. Eur J Cardiothorac Surg, 2003 Apr, 23(4), 473 - 6 Delayed pericardial effusion following stab wounds to the chest; Harris DG et al.; INTRODUCTION: Delayed pericardial effusion following penetrating cardiac trauma has not been commonly reported, and the exact incidence remains unknown . It was more common before 1960, when pericardiocentesis was still a popular treatment for stable patients presenting with a stab wound to the heart . MATERIAL AND METHODS: During an 8-year period, 24 patients were diagnosed with delayed pericardial effusions following a recent stab wound over the chest . Nine patients had been initially treated at our trauma unit, and the remaining 15 patients were referred by a peripheral clinic . RESULTS: Diagnosis was confirmed by cardiac ultrasound or echocardiogram . Sixteen patients were adequately treated by subxiphoid drainage . Sternotomy was performed in five patients, left thoracotomy in two and right thoracotomy in one patient . No actively bleeding injuries were found . Three patients had active infection in the pericardial space . Fever, pleural effusions and ascites were common associated findings . Additional procedures performed included laparotomy for acute abdominal pain in two patients (both negative), and simultaneous drainage of a pleural empyema . Two patients with staphylococcal pericardial infections required subsequent pericardiectomy . SUMMARY: The diagnosis of a penetrating cardiac patient may be missed in a stable patient, and patients may present with delayed pericardial effusions and tamponade . Post pericardiotomy syndrome may be the most common cause of delayed pericardial effusion, followed by sepsis . Subxiphoid pericardial window is an adequate form of treatment . Recent literature reveals that occult cardiac injury is not uncommon, thus a case should be made to actively investigate all patients with precordial stab wounds with cardiac ultrasound or echocardiogram. Biochemistry (Mosc), 2003 Jan, 68(1), 50 - 3 Purification and some properties of lysostaphin, a glycylglycine endopeptidase from the culture liquid of Staphylococcus simulans biovar staphylolyticus; Fedorov TV et al.; This work presents a method for purification of lysostaphin, a glycylglycine endopeptidase, from the culture liquid of S . simulans biovar staphylolyticus to homogeneity in a few steps . The method includes ultrafiltration and ion-exchange and hydrophobic chromatographies . The enzyme was isolated in preparative amounts with the yield of 51% . Some physical and chemical properties of the enzyme are described. Scand J Infect Dis, 2003, 35(2), 110 - 3 Infectious complications after mass disasters: the Marmara earthquake experience; Keven K et al.; The Marmara earthquake occurred on 17 August 1999 . There were 639 renal victims, of whom 477 needed some form of renal replacement therapy . Although several medical complications have been reported in the literature, there has been no detailed description of infectious complications in patients with crush syndrome after earthquakes . Data from 35 hospitals considering clinical and laboratory findings, as well as infectious complications and the results of microbiological examinations, were analysed . 223 out of 639 (34.9%) patients had infectious complications, which comprised the most frequent medical problem in the renal victims . The patients who suffered from infections had a higher mortality rate than those who did not (p = 0.03) . Sepsis and wound infection were the main presentation of the infectious complications . 121 (18.9%) patients suffered from sepsis; the mortality rate was higher in these patients (27.3%) than in victims who did not suffer from sepsis (12.4%, p < 0.0001) . In a multivariate model, sepsis was associated with increased mortality (p = 0.0002, odds ratio 2.45, 95% confidence interval 1.52-3.96) . 53 (8.2%) and 41 (6.4%) patients had wound and pulmonary infections, respectively . Most of the infections were nosocomial in origin and caused by Gram-negative aerobic bacteria and Staphylococcus spp . Infectious complications are common in renal victims of catastrophic earthquakes and are associated with increased mortality when complicated by sepsis. Shock, 2003 Apr, 19(4), 352 - 7 Differential induction of hepatic dysfunction after intraportal and intravenous challenge with endotoxin and Staphylococcal enterotoxin B; Beno DW et al.; We have previously shown that systemic infusion of the bacterial toxins Staphylococcal enterotoxin B (SEB) and endotoxin (LPS) induces hepatic dysfunction as measured by decreased biliary indocyanine green (ICG) excretion . In this study, we compare the effects of these bacterial toxins after infusion into the portal and systemic circulation and directly measure biliary bile acid excretion as a measure of cholestasis . We hypothesized that bacterial toxins infused into the portal vein would induce greater hepatic dysfunction than toxins infused into the systemic circulation . Using a chronically catheterized rat model, biliary bile acid excretion was directly measured after infusion of LPS at 10 and 100 microg/kg with and without 50 microg/kg SEB into the portal vein (IPV) or inferior vena cava (IV) at baseline, and at 6 and 24 h . We found that when LPS was infused alone, only IPV administration caused a significant decrease in bile acid excretion at 6 h . There was no change in bile acid excretion after IV administration of LPS . In contrast, when the combination of LPS and SEB was infused, both IV and IPV administration significantly decreased bile acid excretion at 6 and 24 h . At 6 h post-LPS and -SEB administration, the decrease in bile acid excretion was significantly greater after IPV than IV administration . There was no site-specific difference in IFN-gamma release after infusion of toxins . However, peak TNFalpha release was decreased in IPV-infused rats {10 microg/kg (P < 0.05) or 100 microg/kg (P = ns) LPS with SEB} compared with the same doses in IV-infused rats . These data question the role of systemic TNF-alpha and IFN-gamma in regulating hepatic dysfunction and suggest a differential functional response of the liver to systemic and gut-derived septic events . This study also further explains the frequent development of liver dysfunction in patients with sepsis, multisystem organ failure, and other diseases with altered intestinal permeability. Leuk Lymphoma, 2003 Feb, 44(2), 379 - 80 Polyclonal reactive peripheral blood plasmacytosis mimicking plasma cell leukemia in a patient with Staphylococcal sepsis; Shtalrid M et al.; A 41-year-old man presented with rhabdomyolysis and sepsis while the peripheral blood smear showed a pseudo-leukemic picture of plasma cells . After starting supportive therapy, the morphologic finding disappeared within 24 h. Acta Vet Hung, 2003, 51(1), 61 - 72 Immunological anomalies and thrombocytopenia in 117 dogs and cats diagnosed with chronic fatigue syndrome (CFS); Tarello W; Retrospective analysis of immune dysfunctions found in 55 dogs and 62 cats diagnosed with Chronic Fatigue Syndrome (CFS), revealed leukopenia in 11% of dogs (n = 6) and 22.5% of cats (n = 14), lymphopenia in 14.5% of dogs (n = 8) and 10% of cats (n = 6), hypogammaglobulinaemia in 9% of dogs (n = 5) and 13% of cats (n = 8) and thrombocytopenia in 20% of dogs (n = 11) and 68% of cats (n = 42) . All patients had creatine kinase enzyme levels above the normal range (CK = 5-100 IU/L) and carried micrococcus-like organisms on erythrocytes . Blood cultures proved positive for Staphylococcus spp . in 16 cases . After low-dosage arsenic-based therapy (thiacetarsamide sodium) all animals experienced complete clinical remission . Subsequent controls demonstrated immune restoration in 4 representative FIV-FeLV negative cats, previously diagnosed with CFS associated with leukopenia, lymphopenia, hypogammaglobulinaemia and thrombocytopenia . The main conclusion is that a CFS-like disease in dogs and cats, characterised by the common hallmarks of high CK levels, absence of known causes of chronic fatigue in animals and presence of micrococcus-like organisms in the blood, can be associated with humoral and/or cellular immune deficiencies in 9-22.5% of cases and with thrombocytopenia in 20-68% of cases . Considerations are made on the possible role of micrococci in the aetiology of the condition and on the similarities with CFS in humans. J Clin Microbiol, 2003 Apr, 41(4), 1404 - 9 Staphylococcus lugdunensis infections: high frequency of inguinal area carriage; van der Mee-Marquet N et al.; Following a change in surgical practice, we noted that the rate at which Staphylococcus lugdunensis was isolated from samples from the plastic surgery unit of our hospital increased considerably . We investigated the sources of these S . lugdunensis strains, and we found that in the case of drain colonization or surgical site infection, the strain was more likely to have come from the patient's skin bacteria when the pubic site had been shaved preoperatively . To test the hypothesis of pubic site colonization, we evaluated the prevalence of S . lugdunensis carriage among the cutaneous flora of the inguinal area . We found that 22% of 140 incoming patients carried S . lugdunensis in this area and that carriage at both inguinal folds was frequent (68% of carriers) . A study of the genetic structure of the total population, including the clinical (n = 18) and the commensal (n = 53) strains, revealed that the diversity of the species was low and that the population was composed of two major groups that diverged at a distance of 35% . No particular characteristics made it possible to distinguish between clinical and commensal strains . Only isolates producing beta-lactamase were homogeneous; six of the eight beta-lactamase-positive strains displayed the same pulsed-field gel electrophoresis pattern. Clin Exp Allergy, 2003 Apr, 33(4), 435 - 41 Specific patterns of responsiveness to microbial antigens staphylococcal enterotoxin B and purified protein derivative by cord blood mononuclear cells are predictive of risk for development of atopic dermatitis; Sharp MJ et al.; BACKGROUND: Mononuclear cells from children with active atopic dermatitis (AD) have been reported to be hyper-responsive to certain microbial stimuli, in particular staphylococcal enterotoxin B (SEB) . However, it is not known whether this responsiveness is acquired during disease development, or is inherent . We investigated this question in a cohort of children at high risk of atopy followed prospectively from birth to age 3 years . We asked whether their cord blood mononuclear cell (CBMC) cytokine responses to SEB, to an unrelated microbial stimulus purified protein derivative (PPD), or to common allergens, were predictive of risk for subsequent AD development during infancy . METHODS: Children at high risk of developing atopy were randomly selected from an ongoing prospective cohort . Cord blood was collected at birth . The children were seen at 6 months, 1, 2 and 3 years and examined for the development of AD . IFN-gamma, IL-5, IL-10 and IL-13 production by CBMC cultured in the presence of SEB, PPD, PHA, house dust mite (HDM) allergen, ovalbumin (OVA) and cat allergen was determined . RESULTS: SEB-induced IL-5 production by CBMC was elevated in children who developed AD at 6 months (P = 0.01) and 2 years (P = 0.009) . PPD-induced IL-5 responses were also elevated in CBMC from children who developed AD at 6 months, 2 years and 3 years (P = 0.05, P = 0.06 and P = 0.06, respectively), as were PPD-induced IL-10 responses (P = 0.05 at 1 years, P = 0.007 at 2 years, P = 0.003 at 3 years) and corresponding IFN-gamma responses (P = 0.05 at 6 months, P = 0.003 at 2 years, P = 0.0004 at 3 years) . Increased IL-10 responses to HDM allergen were also observed throughout the observation period in CBMC from children who developed AD . CONCLUSION: Children who develop infantile AD appear to have a predisposition to respond to SEB in a Th2-dominant manner involving selective stimulation of IL-5 production . The increased IL-10 and IFN-gamma induced in response to PPD by children with AD may point to additional intrinsic differences in responses to microbial stimuli between those at high vs . those at low risk for AD, which merit more detailed investigations. J Recept Signal Transduct Res, 2003 Feb, 23(1), 33 - 52 A serine/threonine phosphorylation site in the ectodomain of a T cell receptor beta chain is required for activation by superantigen; Lukashev DE et al.; The presence of consensus phosphorylation sites in the ectodomains of cell surface proteins suggests that such post-translational modification may be important in regulation of surface receptor activity . To date, the only cell surface receptor for which such ectodomain phosphorylation has been conclusively demonstrated is the clonally expressed T cell antigen receptor (TCR) . Attempts to conclusively identify individual phosphorylated residues in TCR alpha and beta chains and determine their functional significance by biochemical approaches failed due to insufficient quantities of purified molecules . Here we present the results of an alternative approach where survey of phosphorylation sites in the TCR alpha and beta chains was accomplished using site-directed mutagenesis and retroviral vector expression, as well as in vitro phosphorylation of synthetic peptide substrates . All mutants studied directed the cell surface expression of normal amounts of TCR, and all transfectants could be stimulated to produce IL-2 in response to substrate-immobilized antibody to TCR . However, mutation of serine-88 in the protein kinase A phosphorylation site of the TCR beta chain resulted in a complete lack of response to the superantigen staphylococcal enterotoxin B (SEB) . In addition, this mutation abolished TCR-associated tyrosine phosphorylation, consistent with the impairment of cell signaling . Reversion of the serine-88/alanine mutation with phosphorylatable threonine completely restored the SEB recognition by TCR . These results, interpreted in the context of the known three-dimensional structure of the complex of SEB and TCR, are consistent with the view that serine-88 is important for the contact of the TCR beta chain with SEB. Eur J Pediatr, 2003 Jun, 162(6), 406 - 9 Epub 2003 Mar 27. Hepatic abscesses associated with umbilical catheterisation in two neonates; Moens E et al.; We describe two neonates with a liver abscess after umbilical venous catheterisation . The first case was a female neonate, born at 32 weeks of gestation . After persistence of elevated inflammatory parameters, an abscess in the right lobe of the liver was diagnosed . Percutaneous drainage under CT guidance was performed . The aspirated pus grew Staphylococcus epidermidis . Inflammatory parameters normalised after 27 days of antimicrobial therapy (vancomycin, cefotaxim, rifampicin) . The second case was in a male neonate, born at 29 weeks of gestation . Percutaneously aspirated pus from the liver abscess was cultured and remained sterile . The patient received antimicrobial therapy (vancomycin, cefotaxim, amikacin) for 26 days and was cured with conservative treatment . CONCLUSION: hepatic abscess should be considered in any infant with an umbilical catheter-associated sepsis and persistent inflammatory response in spite of adequate antimicrobial therapy, especially when signs of abdominal infection are present. J Mol Biol, 2003 Apr 18, 328(1), 255 - 71 Phase diagrams: a graphical representation of linkage relations; Rosgen J et al.; It is shown here that phase diagrams of ligand-binding biological macromolecules provide a powerful tool for the analysis of reaction mechanisms . The present study provides simple rules for the construction and interpretation of such phase diagrams . We give examples for the derivation of reaction schemes for macromolecules that can bind two different kinds of ligands . By sampling one dimension of a phase diagram it is possible to reconstruct the second dimension, including the correct stoichiometry, positive and negative linkage between the ligands and equilibrium binding constants for the complete series of reactions . The discussion is generalised to temperature and pressure-dependent phase diagrams . To exemplify the new diagram method we analyse the pH-dependent binding of trans-beta-indole acrylic acid to apo-Trp repressor, the pH-dependent thermal denaturation of alpha-chymotrypsinogen A, calcium binding and denaturation of annexin I, high affinity zinc binding to a metallo-beta-lactamase and high-pressure and temperature denaturation of RNase A and staphylococcal nuclease . Protein Pept Lett, 2003 Apr, 10(2), 175 - 81 An efficient fusion expression system for protein and peptide overexpression in Escherichia coli and NMR sample preparation; Cheng Y et al.; An efficient fusion expression system with a small fusion partner, His6-tagged N-terminal fragment of staphylococcal nuclease R, has been constructed and tested with two genes . The results show that the system is not only suitable for overexpression of small proteins and peptides but simplifies purification of target proteins and peptides . The study also provides a practical method for preparation of isotope-labeled protein sample for NMR analysis. J Mol Graph Model, 2003 Jun, 21(6), 473 - 86 Prediction of the multimeric assembly of staphylococcal enterotoxin A with cell-surface protein receptors; Cuff L et al.; Staphylococcal enterotoxin A (SEA) cross-links two class II major histocompatibility complex (MHC) molecules and forms a multimeric assembly with T-cell receptors (TcRs) . The X-ray crystal structure of SEA has been solved, yet details describing molecular recognition and association remain unclear . We present a structural model for the interactions of SEA with cell-surface proteins . Molecular docking calculations predicting SEA association with the class II MHC molecule HLA-DR1 were performed by using a rigid-body docking method . Docked orientations were evaluated by a Poisson-Boltzmann model for the electrostatic free energy of binding and the hydrophobic effect calculated from molecular surface areas . We found that the best-scoring SEA conformers for the DR1alpha interface display a binding mode similar to that determined crystallographically for staphylococcal enterotoxin B bound to HLA-DR1 . For the zinc-binding site of SEA, docking DR1beta yielded several orientations exhibiting tetrahedral-like coordination geometries . Combining the two interfaces, tetramers were modeled by docking an alphabeta TcR with trimolecular complexes DR1beta-SEA-DR1alpha and SEA-betaDR1alpha-SEA . Our results indicate that the complex DR1beta-SEA-DR1alpha provides a more favorable assembly for the engagement of TcRs, forming SEA molecular contacts that are in accord with reported mutagenesis studies . In contrast, the cooperative association of two SEA molecules on a single DR1 molecule sterically inhibits interactions with TcRs . We suggest that signal transduction stimulated by SEA through large-scale assembly is limited to four or five TcR-(DR1beta-SEA-DR1alpha) tetramers and requires the dimerization of class II MHC molecules, while TcR dimerization is unlikely. J Perinatol, 2003 Mar, 23(2), 94 - 7 Use of oxygen cannulas in extremely low birthweight infants is associated with mucosal trauma and bleeding, and possibly with coagulase-negative staphylococcal sepsis; Kopelman AE et al.; OBJECTIVE: We studied the association between the use of oxygen cannulas (OCs) and (1) nasal bleeding and (2) coagulase-negative staphylococcal sepsis (CNSS) . STUDY DESIGN: Review of care sheets, with chi(2) or sign-test group comparisons . RESULTS: Infants treated with OCs were suctioned more frequently (2.6 vs 1.3 times per day, p<0.001), and had more bloody nasal secretions (34.6% vs 4.6%, p<0.05) that increased with increasing OC days . By 10 days, 90% of infants had experienced bloody secretions.CNSS occurred less often in infants treated with oxyhoods than those on OC or CPAP (1 of 13, 8%, vs 10 of 44, 23%), but the difference was not significant . Eight of the 10 CNSS episodes clustered within 3 and 7 days of starting CPAP or cannula treatments . CONCLUSION: OC use in extremely low birthweight infants is associated with nasal mucosal injury and bleeding . Studies are needed to see if use of OCs is a risk factor for CNSS. Eur J Immunol, 2003 Apr, 33(4), 970 - 9 The CD4-related molecule, LAG-3 (CD223), regulates the expansion of activated T cells; Workman CJ et al.; The lymphocyte activation gene-3 (LAG-3, CD223) is a CD4-related, activation-induced cell surface molecule that binds to MHC class II with high affinity . The function of murine LAG-3 on T cells is unclear . Here, we show that V beta 7/8(+)LAG-3(-/-) T cells expand poorly following staphylococcal enterotoxin B (SEB) stimulation in vitro . LAG-3(-/-) T cells proliferate at a normal rate, but exhibit increased cell death . Similar observations were made with LAG-3(-/-)CD4(+)OT-II TCR transgenic T cells following peptide stimulation . Despite reduced T cell expansion and increased cell death, LAG-3(-/-)OT-II(+) T cells secrete more IL-2 and IFN-gamma following stimulation . Antigen-driven expansion of LAG-3(-/-) T cells was restored by constitutive expression of LAG-3 via retroviral-mediated stem cell gene transfer . We further show that LAG-3 function is mediated via its cytoplasmic domain, for which a conserved 'KIEELE' motif is essential . Our data support a role for LAG-3 in regulating the expansion of activated T cells. Pediatrics, 2003 Apr, 111(4 Pt 2), e519 - 33 Implementation of evidence-based potentially better practices to decrease nosocomial infections; Kilbride HW et al.; OBJECTIVE: Six neonatal intensive care units (NICUs) that are members of the Vermont Oxford National Evidence-Based Quality Improvement Collaborative for Neonatology collaborated to reduce infection rates . There were 7 centers in the original focus group, but 1 center left the collaborative after 1 year . Nosocomial infection is a significant area for improvement in most NICUs . METHODS: Six NICUs participating in the Vermont Oxford Network made clinical changes to address 3 areas of consensus: handwashing, line management, and accuracy of diagnosis . The summary statements were widely communicated . Review of the literature, internal assessments, and benchmarking visits all contributed to ideas for change . RESULTS: The principle outcome was the incidence of coagulase-negative staphylococcus bacteremia . There was an observed reduction from 24.6% in 1997 to 16.4% in 2000 . CONCLUSIONS: The collaborative process for clinical quality improvement can result in effective practice changes. Biochem Biophys Res Commun, 2003 Apr 11, 303(3), 863 - 7 A very sensitive enzyme-linked immunoabsorbent assay to Staphylococcal protein A in the presence of immunoglobulins; Valdes Veliz R et al.; The production of recombinant hepatitis B virus surface antigen (rHBsAg) purified by immunoaffinity chromatography with monoclonal antibodies is used to obtain a vaccine against this virus . Monoclonal antibodies to rHBsAg from mouse ascites have been purified by Staphylococcal Protein A (SpA)--prior coupling to Sepharose CL-4B (Amersham-Bioscences, Uppsala, Sweden) . A high sensitivity immunoassay has been developed for the quantification of part-per-million of SpA contaminants likely to co-purify with monoclonal antibodies obtained by Protein A affinity chromatography, in the presence of immunoglobulins . Specific sheep polyclonal Abs against SpA (SpAc1) were used as plate coating and the SpA detection was possible thanks to the conjugates of sheep Ab fragments F(ab)(2) (fSpAc1) and horseradish peroxidase (fSpAc1-peroxidase), reducing the possible unspecific interaction between SpA and Fc fragments . The immunoassay was shown to be specific for SpA contaminants . The quantification limit of the assay was 0.39 ng/ml spreading to the measurement of contamination levels less than 2 ppm of SpA in final preparations of monoclonal antibodies used for the immunopurification of pharmaceutical products, which is quite low for this application. Cytokine, 2003 Jan 21, 21(2), 65 - 73 Regulation of Staphylococcus epidermidis-induced IFN-gamma in whole human blood: the role of endogenous IL-18, IL-12, IL-1, and TNF; Stuyt RJ et al.; Interleukin 12 (IL-12) and IL-18 act synergistically to stimulate interferon gamma (IFN-gamma) production; moreover, IL-1 and tumor necrosis factor (TNF) may also augment IFN-gamma synthesis . We have investigated the relative contributions of these cytokines in the production of IFN-gamma and TNF by the Gram-positive bacterium Staphylococcus epidermidis, using the specific cytokine inhibitors IL-18 binding protein (IL-18BP), IL-1 receptor antagonist (IL-1Ra), anti-IL-12 antibodies (anti-IL-12 Ab), and TNF binding protein . Inhibition of caspase-1 reduced IFN-gamma and IL-1beta levels (by 80 and 67%, respectively) when heat-killed S . epidermidis was added to whole human blood cultures . IL-18BP reduced S . epidermidis-induced IFN-gamma (77% maximal suppression) . In contrast, blocking IL-1 receptors by IL-1Ra had no effect on IFN-gamma production . Blocking endogenous IL-12 and TNF reduced IFN-gamma production by 69 and 36% . S . epidermidis-induced TNF-alpha was inhibited by IL-18BP and IL-1Ra, but not anti-IL-12 Ab, whereas IL-8 production was unaffected by any of the specific cytokine blocking agents . In conclusion, S . epidermidis stimulates IFN-gamma which is IL-18, IL-12 and TNF-dependent, but IL-1 independent. Biophys J, 2003 Apr, 84(4), 2533 - 41 Reorientational dynamics of enzymes adsorbed on quartz: a temperature-dependent time-resolved TIRF anisotropy study; Czeslik C et al.; The preservation of enzyme activity and protein binding capacity upon protein adsorption at solid interfaces is important for biotechnological and medical applications . Because these properties are partly related to the protein flexibility and mobility, we have studied the internal dynamics and the whole-body reorientational rates of two enzymes, staphylococcal nuclease (SNase) and hen egg white lysozyme, over the temperature range of 20-80 degrees C when the proteins are adsorbed at the silica/water interface and, for comparison, when they are dissolved in buffer . The data were obtained using a combination of two experimental techniques, total internal reflection fluorescence spectroscopy and time-resolved fluorescence anisotropy measurements in the frequency domain, with the protein Trp residues as intrinsic fluorescence probes . It has been found that the internal dynamics and the whole-body rotation of SNase and lysozyme are markedly reduced upon adsorption over large temperature ranges . At elevated temperatures, both protein molecules appear completely immobilized and the fractional amplitudes for the whole-body rotation, which are related to the order parameter for the local rotational freedom of the Trp residues, remain constant and do not approach zero . This behavior indicates that the angular range of the Trp reorientation within the adsorbed proteins is largely restricted even at high temperatures, in contrast to that of the dissolved proteins . The results of this study thus provide a deeper understanding of protein activity at solid surfaces. Clin Chem Lab Med, 2003 Feb, 41(2), 159 - 63 Biliary bacteria in living related liver transplant recipients: microbiology and rapid detection system using flow cytometry; Saito T et al.; Since cholangitis is a major complication in living related liver transplant (LRLT) recipients, rapid detection of biliary bacteria is necessary for the management of patients . We have developed a screening method for the detection of biliary bacteria using flow cytometry (FCM) . Two hundred and seventy eight bile samples were obtained from 50 patients with biliary drainage tubes after LRLT at Kyoto University Hospital between July and September 2001 . Of the 278 samples, 165 (59.3%) were culture-positive . The most common isolates were Enterococcus species, Pseudomonas species, Staphylococcus species, Klebsiella species, and Candida species . As the original FCM system was inadequate for specifically detecting bacteria in bile samples, we established the most appropriate gate and cut-off value from the particle distribution represented on scattergram of the forward-scattered light and fluorescent light intensity . The 3% cut-off value was most preferably related to the culture results . The FCM system detected biliary bacteria with a sensitivity of 93.9%, specificity of 81.4%, positive predictive value of 88.1%, negative predictive value of 90.2%, false-positive rate of 7.6%, false-negative rate of 3.6%, and percent agreement of 88.9% between FCM and culture . Therefore, FCM can be a useful method in clinical laboratories for the rapid screening for biliary bacteria in LRLT recipients. Int J Pediatr Otorhinolaryngol, 2003 Apr, 67(4), 409 - 12 Hyper immunoglobulin-E syndrome: a case with chronic ear draining mimicking polypoid otitis media; Gorur K et al.; The hyper-IgE syndrome is a rare, complex immunoregulatory disorder characterized by pruritic dermatitis, recurrent staphylococcus skin abscesses and extremely elevated levels of IgE in serum . In this report, an 8-year-old girl with hyper-IgE syndrome is presented . She had pruritic dermatitis and skin abscesses on her extremities and scalp . Discharge and granulation tissue in right external ear canal were recognized in otorhinolaryngologic examination . Cultures of the suppuration of the external ear canal yielded S . aureus . Biopsy of the lesion was reported as granulation tissue . Local treatment of the ear canal was performed with ear wick soaked with steroid and antibiotic solutions two times per day. Ann Pharmacother, 2003 Apr, 37(4), 517 - 20 Mechanisms for linezolid-induced anemia and thrombocytopenia; Bernstein WB et al.; BACKGROUND: Linezolid has been associated with anemia and thrombocytopenia . Mechanisms for neither have been elucidated . OBJECTIVE: To propose mechanisms for linezolid-induced anemia and thrombocytopenia . CASE SUMMARY: A 78-year-old white woman with Staphylococcus epidermidis endocarditis was treated with linezolid after developing resistance to multiple antibiotic regimens . After 7 days of linezolid therapy, she developed thrombocytopenia, while an anemia present since admission remained unchanged . A bone marrow biopsy was performed, primarily looking for a mechanism for the thrombocytopenia . Histopathology revealed adequate megakaryocytes, ringed sideroblasts, and vacuolated pronormoblasts . A course of immune globulin (IVIG) was administered, with slowing in the rate of decline in platelets . She died 24 hours after her last dose of IVIG of congestive heart failure . DISCUSSION: The presence of ringed sideroblasts and vacuolated pronormoblasts suggests that linezolid-induced anemia is secondary to a chloramphenicol-like suppression of erythropoiesis . The presence of adequate, normal-appearing megakaryocytes suggests immune-mediated thrombocytopenia, not marrow suppression . Although the response to IVIG is difficult to interpret because of the patient's death, there was a slowing in the rate of decline of the platelet count, further supporting immune-mediated thrombocytopenia . An objective causality assessment indicated that the adverse drug event was probably due to linezolid . CONCLUSIONS: There appear to be 2 distinct mechanisms for linezolid-induced cytopenias . While anemia is reversible and manageable with transfusions, thrombocytopenia can be a treatment-limiting toxicity . The ability to treat through an immune-mediated cytopenia with IVIG may be beneficial for critically ill patients with few therapeutic options. Semin Respir Infect, 2003 Mar, 18(1), 17 - 22 Does infection play a role in the pathogenesis of pulmonary vasculitis? Capizzi SA, Specks U. The pulmonary vasculitides are a heterogeneous group of systemic inflammatory diseases of unknown etiology with potential for significant morbidity . The syndromes with particular predilection for the respiratory tract are Wegener's granulomatosis, microscopic polyangiitis, and Churg-Strauss syndrome . The discovery of antineutrophil cytoplasmic antibodies (ANCA) in these disorders has facilitated their diagnosis and contributed to the understanding of their pathogenesis . Clinical studies and some animal models suggest a disease-modifying role for antimicrobial therapy in ANCA-associated vasculitis . Nasal colonization with Staphylococcal aureus is an independent risk factor for relapse of Wegener's granulomatosis . This evidence suggests infectious pathogens as potential triggers of a cascade of events that result in vascular inflammation . Multiple laboratory studies have contributed to a coherent and plausible theory about the pathogenesis of ANCA-associated vasculitis in which infection plays a critical role . In susceptible individuals immune tolerance may break down and ANCA production resulting from molecular mimicry ensues . In addition, bacterial superantigens may serve as potent stimulators of the immune system . In this context, ANCA directed against proteinase 3 or myeloperoxidase may interact with their target antigens expressed on the surface of activated neutrophils, leading to an enhanced and perpetuated inflammation of vessels . Despite significant advances, the precise connection between infections and pulmonary vasculitis remains poorly understood, and further studies into the pathogenesis of these diseases are needed . J Chromatogr B Analyt Technol Biomed Life Sci, 2003 Mar 25, 786(1-2), 319 - 25 GST-Fbe can recognize beta-chains of fibrin(ogen) on explanted materials; Pei L et al.; Staphylococcus epidermidis, a coagulase-negative staphylococcus (CoNS), is one of the leading pathogens of nosocomial infections, particularly associated with foreign body infections . Adherence of S . epidermidis to fibrinogen deposited on the surfaces of implants is important for the development of foreign body infections . A gene (fbe) encoding a fibrinogen-binding protein from S . epidermidis (Fbe) was identified by shotgun phage display . A portion of fbe was cloned into a GST-fusion vector . Affinity to glutathione-Sepharose by the GST-tag and affinity to fibrinogen-Sepharose by the Fbe part were applied to purify the recombinant Fbe . The purity and efficacy of the methods used in protein purification was compared . Furthermore, the potential physiological role of Fbe was studied by the interaction between GST-Fbe and components extracted from explanted materials in vitro. AIDS Treat News, 2003 Feb 7, (388), 2 - 3 Antibiotic-resistant skin infections spreading among gay men, also in prisons; James JS; An outbreak of staphylococcus skin infection, causing large boils that are difficult to treat because these bacteria are resistant to many antibiotics, has spread recently among gay men in some cities, and also in certain prisons . In the past this infection occurred mainly in hospitals. J Cataract Refract Surg, 2003 Feb, 29(2), 371 - 8 In vitro study of bacterial adherence to processed dura mater, processed pericardium, pericardium in saline, and human sclera; John T et al.; PURPOSE: To study bacterial adherence to processed dura mater, processed pericardium, pericardium in saline, and human sclera and the difference in bacterial adherence to these tissues . SETTING: Research Laboratory, Loyola University Medical Center, Maywood, Illinois, USA . METHODS: Specimens of processed dura mater, processed human pericardium, pericardium in saline, and human sclera (N = 32) were exposed to Staphylococcus epidermidis (concentration 3 x 10(8)) for 10, 20, 40, and 60 minutes, washed for 5 seconds, fixed, and processed for scanning electron microscopy (SEM) . Each bacterial count represents an average of 50 random SEM fields at x5,000 magnification . After SEM, selected specimens were processed for transmission electron microscopy . RESULTS: The mean number of bacteria/mm(2) +/- SD adhering to the tissues at 10, 20, 40, and 60 minutes, respectively, were dura mater, 107,833 +/- 65,410, 104,500 +/- 13,471, 96,067 +/- 113,884, and 204,267 +/- 153,697; processed pericardium, 131,550 +/- 86,194, 100,900 +/- 20,031, 144,683 +/- 51,730, and 176,933 +/- 111,818; pericardium in saline, 7,925 +/- 1,520, 33,933 +/- 32,085, 1,217 +/- 1,287, and 21,550 +/- 16,436; and human sclera, 4,850 +/- 2,121, 23,700 +/- 17,961, 5,150 +/- 1,273, and 8,175 +/- 8,450 . A 2-way analysis of variance showed significant differences among groups (P =.001) and no significant difference in sample time (P =.929) . CONCLUSIONS: Bacterial adherence to processed dura mater, processed pericardium, pericardium in saline, and human sclera should be considered when these materials are used clinically during ophthalmic surgery and other surgical specialties . Adequate broad-spectrum antibiotic coverage is needed to prevent infection and subsequent complications in patients. J Cataract Refract Surg, 2003 Feb, 29(2), 361 - 6 Bacterial adhesion to rigid and foldable posterior chamber intraocular lenses: in vitro study; Schauersberger J et al.; PURPOSE: To determine the in vitro bacterial adherence to intraocular lenses (IOLs) routinely used at 1 center . SETTING: University Hospital of Vienna, Vienna, Austria . METHODS: Four types of rigid IOLs (hydrophobic poly{methyl methacrylate} {PMMA} and hydrophilic heparin-surface-modified PMMA) and 5 types of foldable intraocular lenses (hydrophobic silicone, hydrophobic acrylic, and hydrophilic acrylic) were used in the experiment . Under standardized conditions, the IOLs were contaminated with 2 strains of Staphylococcus epidermidis . A sonication method and impression method were used for quantification . The mean bacterial density per lens type (colony forming units/mm(2)) was compared . RESULTS: With both germs and both quantification methods, more bacteria was found on hydrophobic lenses than on IOLs with hydrophilic surfaces (P =.001) . The Alcon AcrySof and Askin UV80F IOLs had the greatest and the Corneal Acrygel and Bausch & Lomb Hydroview IOLs the least affinity to these microorganisms . CONCLUSION: The data suggest that hydrophilic IOLs can help reduce the rate of postoperative endophthalmitis as a result of their surface properties. J Dairy Sci, 2003 Feb, 86(2), 530 - 7 Influence of pulsation rate on udder health and teat thickness changes in dairy ewes; Peris C et al.; In this work on machine milking of ewes, pulsation rates of 120 and 180 cycles per min were compared, both with a pulsation ratio of 50:50 and a vacuum level of 36 kPa, comparing intramammary infection (IMI), somatic cell count (SCC) and teat end thickness changes . To this end, two groups of 20 Manchega ewes were used in a crossover experimental design with two experimental periods of 24 d for each . Bacterial exposure of all teats was increased by dipping them in a suspension of Staphylococcus simulans at four consecutive milkings of each period . Pulsation rate of 180 cycles per min, compared with 120 cycles per min, had no negative effect upon new IMI (11 and 16% of ewes infected, respectively) and SCC . No teat end lesions were observed in those animals milked with the two pulsations assayed . Also, teat thickness changes (-0.38 and -0.36 mm at 120 and 180 cycles per min, respectively) were not affected significantly . Finally, in absence of IMI, the two pulsation rates assayed did not affect the SCC. Biophys Chem, 2003, 100(1-3), 261 - 80 Addition of side chains to a known backbone with defined side-chain centroids; Kazmierkiewicz R et al.; An automatic procedure is proposed for adding side chains to a protein backbone; it is based on optimization of a simplified energy function for peptide side chains, given its backbone and positions of side-chain centroids . The energy is expressed as a sum of the energies of interaction between side chains, and a harmonic penalty function accounting for the preservation of the positions of the C(alpha) atoms and the side-chain centroids . The energy of side-chain interactions is calculated with the soft-sphere ECEPP/3 potential . A Monte Carlo search is carried out to explore all possible side-chain orientations within a fixed backbone and side-chain centroid positions . The initial, usually extended, side-chain conformations are taken directly from the ECEPP/3 database . The procedure was tested on six experimental (X-ray or NMR) structures: immunoglobulin binding protein (PDB code 1IGD, an alpha+beta-protein); transcription factor PML (PDB code 1BOR, a 49-104 fragment of the ring finger domain, predominantly beta-protein); bovine pancreatic trypsin inhibitor (crystal form II) (PDB code 1BPI, an alpha+beta-protein); the monomer of human deoxyhemoglobin (PDB code 1BZ0, an alpha-helical structure); chain A of alcohol dehydrogenase from Drosophila lebanonensis (PDB code 1A4U); as well as on the 10-55 portion of the B domain of staphylococcal protein A (PDB code 1BDD) . In all cases except 1BPI, the data for the algorithm (i.e . the backbone or C(alpha) coordinates and the positions of side-chain centroids) were taken from the experimental structures . For protein A, the C(alpha) coordinates and positions of side-chain centroids were also taken from the 1.9-A-resolution model predicted by the UNRES force field . In all comparisons with experimental structures, complete side-chain geometry was reconstructed with a root-mean-square (RMS) deviation of approximately 0.6-0.9 A from the heavy atoms when complete backbone and side-chain-centroid coordinates were used in reconstruction, or approximately 1.0 A when the C(alpha) and centroid coordinates were used. Eur J Ophthalmol, 2003 Jan-Feb, 13(1), 11 - 7 Does identification of the causal organism of corneal ulcers influence the outcome? Pharmakakis NM, Andrikopoulos GK, Papadopoulos GE, Petropoulos IK, Kolonitsiou FI, Koliopoulos JX. PURPOSE: To investigate whether identification of the causal organism in corneal ulcers influences their outcome . METHODS: We retrospectively studied 114 patients, 72 males and 42 females aged 6-89 years, admitted to this eye clinic during the years 1994-2000 on account of an infectious corneal ulcer . Their examination included a detailed history, visual acuity measurement, and biomicroscopy in everyday follow-up . The ulcers were classified according to their severity and outcome . We assessed the cases where cultures had been done, reviewed the results, and searched for a possible correlation between the outcome and the fact of culturing the ulcer and identifying the causal organism . RESULTS: Of the 114 corneal ulcers studied, 23 were mild, 49 moderate, and 42 severe . Fifty (44%) had not been cultured, but 64 ulcers (56%) had been cultured, with a positive result in 37 cases (58%), Staphylococcus and Pseudomonas species being the most common organisms found . In moderate and severe ulcers, there was a tendency to a higher proportion of successful outcome for cultured ulcers, but with no significant correlation . CONCLUSIONS: Despite a tendency towards favorable results in culture-positive corneal ulcers, the influence of the detection of the organism on their outcome has not been proved . The role of the initial broad-spectrum antibiotic therapy remains important. Tunis Med, 2002 Sep, 80(9), 509 - 14 {Pacemaker infections}; Mokaddem A et al.; Infectious complications following pacemaker implantation are not common but can be particularly severe . The reported incidence varies from 0.5 to 5% in the literature . The duration of the procedure and repeat procedures are considered to be predisposing factors . The main cause of these infections is thought to be local contamination during the implantation . The commonest causal organism is staphylococcus . Because the presentation may be atypical, the diagnosis is often confirmed by transesophageal echocardiography which is the investigation of choice for imaging a vegetation on an endocavitary pacing lead in cases of infectious endocarditis . The seriousness of this infection requires early diagnosis and adapted treatment including double bactericidal antibiotherapy and complete ablation of the material . Systematic preoperative prophylactic antibiotic therapy is recommended. Arthritis Rheum, 2003 Mar, 48(3), 642 - 50 Bacterial peptidoglycans but not CpG oligodeoxynucleotides activate synovial fibroblasts by toll-like receptor signaling; Kyburz D et al.; OBJECTIVE: To test the hypothesis that bacterial products acting as adjuvants, such as CpG oligodeoxynucleotides (ODNs) and peptidoglycans (PGs), are able to activate synoviocytes, and to determine the involvement of Toll-like receptors (TLRs) in this activation process . METHODS: Cultured synovial fibroblasts obtained from patients with rheumatoid arthritis (RA) or osteoarthritis (OA) were stimulated with CpG ODNs or PGs . The expression of various integrins was determined by fluorescence-activated cell sorting . TLR and matrix metalloproteinase (MMP) messenger RNA (mRNA) was measured by real-time polymerase chain reaction . Additionally, levels of interleukin-6 (IL-6) and IL-8 in the culture supernatants were assessed by enzyme-linked immunosorbent assay . Blocking experiments were performed by adding anti-TLR-2 and anti-TLR-4 monoclonal antibodies to cultures stimulated with bacterial PGs . RESULTS: Incubation of synovial fibroblasts with CpG ODNs resulted in neither up-regulation of the expression of integrins on the cell surface, up-regulation of MMP mRNA expression, nor IL-6 and IL-8 production . However, incubation of RA synovial fibroblasts as well as OA synovial fibroblasts with staphylococcal PGs led to an up-regulation of CD54 (ICAM-1) surface expression and to increased expression of MMP-1, MMP-3, and MMP-13 mRNA . Furthermore, production of the proinflammatory cytokines IL-6 and IL-8 was increased by treatment with PGs . We demonstrated that cultured synovial fibroblasts express low levels of TLR-2 and TLR-9 mRNA . TLR-2 was up-regulated after stimulation with PGs, whereas TLR-9 mRNA remained at baseline levels after stimulation with CpG ODNs . Anti-TLR-2 monoclonal antibodies significantly inhibited production of IL-6 and IL-8 induced by stimulation with PGs . CONCLUSION: We demonstrate that bacterial PGs activate synovial fibroblasts, at least partially via TLR-2, to express integrins, MMPs, and proinflammatory cytokines . Inhibition of TLR signaling pathways might therefore have a beneficial effect on both joint inflammation and joint destruction. Euro Surveill, 1997 Apr, 2(4), 25 - 28 Widespread dissemination of epidemic MRSA in German hospitals; Witte W et al.; Strains of Staphylococcus aureusresistant to multiple antibiotics, including those resistant to methicillin (MRSA), present a major problem in the control of hospital acquired infections . Some strains show a definite capacity for spread once introduced in South Med J, 2003 Feb, 96(2), 194 - 7 Panhemispheric infarction: a complication of cuffed catheter; Agraharkar M et al.; The need for reliable vascular access remains the Achilles heel of hemodialysis . Complications of vascular access are a leading cause of morbidity and mortality in patients who undergo hemodialysis, especially in those patients with end-stage renal disease . Among methods of vascular access, arteriovenous fistulae have the lowest rate of infection and should be the access of choice when vascular anatomy permits . Also, the incidence of staphylococcal infections in patients infected with human immunodeficiency virus is increasing . To emphasize the need to use arteriovenous fistula access for hemodialysis whenever possible, we report the case of a patient with end-stage renal disease and human immunodeficiency virus infection who died as a result of panhemispheric infarction and uncal herniation as a result of fulminant staphylococcal bacteremia caused by central venous catheter sepsis. Curr Mol Med, 2003 Mar, 3(2), 127 - 38 Atopic dermatitis: molecular mechanisms, clinical aspects and new therapeutical approaches; Galli E et al.; Atopic dermatitis (AD) is a genetically determinated, chronic inflammatory skin disorder associated with cutaneous erythema and severe pruritus, affecting 10-15% of children with increasing incidence and socio-economical relevance . Frequently, AD is associated with development of allergic rhinitis and/or asthma later in childhood . In most of patients AD is associated with a sensitization to food and/or environmental allergens and increased serum-IgE, while only a fewer percentage missed links to the classical atopic diathesis . Currently investigated pathogenetic aspects of AD include imbalanced Th1/Th2 responses, altered prostaglandin metabolism, intrinsic defects in the keratinocyte function, delayed eosinophil apoptosis, and IgE-mediated facilitated antigen presentation by epidermal dendritic cells . An inflammatory response of the two-phase-type and the effects of staphylococcal superantigens (SAgs) are also reported . At present a standardized cure of AD and a consensus on therapeutical approach of the severe form of the disease have not been established . Current management of AD is directed to the reduction of cutaneous inflammation and infection, mainly by S . aureus, and to the elimination of exacerbating factors (irritants, allergens, emotional stresses) . Since patient with AD show abnormalities in immunoregulation, therapy directed to adjustment of their immune function could represent an alternative approach, particularly in the severe form of the disease . In this review, we analyse the clinical and genetic aspects of AD, the related molecular mechanisms, and the immunobiology of the disease, focusing our attention on current treatments and future perspectives on this topic. Zh Mikrobiol Epidemiol Immunobiol, 2003 Jan-Feb, (1), 74 - 7 {Effectiveness of the oral administration of tomicide in experimental infection}; Blinkova LP et al.; Experimental data on the oral administration of Tomicid using different schemes for the protection of white mice from staphylococcal infection are presented . The use of Tomicid, administered in the maximum dose admissible for mice, ensured the protection of up to 2/3 of the total number of mice . A single oral administration of the preparation immediately after infection protected 1/3 of the survived mice from local staphylococcal infection . Good prospects of using Tomicid for the prevention of catarrhal diseases in guinea pigs were established. Lijec Vjesn, 2002 Oct, 124(10), 310 - 3 {Endocarditis associated with atrial and ventricular cardiac pacing leads . Case report}; Bencic ML et al.; The infection of a transvenous lead implanted for cardiac stimulation is a rare, but serious complication . We report observation of a 25-year old man whose Staphylococcus epidermidis sepsis linked to endocarditis was related to atrial and ventricular pacing leads, and was diagnosed after two months of medical treatment . The most important role during the diagnostic process was played by the echocardiographic examination, especially transoesophageal, which revealed the large vegetations on atrial as well as ventricular pacing lead . The diagnosed condition was treated by complete removal of pacing system using open chest surgery and cardiopulmonary pump . After four weeks of vigorous antibiotic treatment, a new DDDR pacing system was implanted, but with epicardial leads. Ann Vasc Surg, 2003 Mar, 17(2), 156 - 61 Epub 2003 Mar 06. Postoperative complications of carotid patching: pseudoaneurysm and infection; Borazjani BH et al.; Patch angioplasty after carotid endarterectomy has been advocated to improve results by decreasing the incidence of recurrent stenosis and postoperative carotid thrombosis . Aneurysmal and infectious complications may be secondary to use of prosthetic materials in arterial reconstruction . We report four patients who developed late operative site complications related to carotid patching . All four of our patients had delayed pseudoaneurysms and three had infection related to the Dacron patch . In a typical case, a 57-year-old who had a right carotid endarterectomy in 1994 presented with a 1-month history of an enlarging right neck mass 7 years later . Imaging revealed a 6 x 4 cm pseudoaneurysm originating from an opening between the patch graft and the old endarterectomized carotid wall . Analysis of the literature disclosed an additional 45 patients who had pseudoaneurysms and/or infection related to carotid patching, most frequently with Dacron . We postulate that a low-grade Staphylococcus epidermidis infection of the foreign body patch may be the etiology . Autogenous saphenous vein interposition graft and antimicrobials effective against gram-positive organisms corrected the pseudoaneurysm . Although the benefits of routine carotid patching may include a decrease in restenosis, this advantage must be weighed against the risk of late pseudoaneurysm and/or infection when a prosthetic patch is used to closed the endarterectomy site. J Antimicrob Chemother, 2003 Mar, 51(3), 585 - 91 Antibacterial poly(D,L-lactic acid) coating of medical implants using a biodegradable drug delivery technology; Gollwitzer H et al.; OBJECTIVES: Biomaterial-associated bacterial infections present common and challenging complications with medical implants . The purpose of this study was to determine the antibacterial properties of a low molecular weight biodegradable poly(D,L-lactic acid) coating with integrated antibiotics gentamicin and teicoplanin . METHODS: Coating of Kirschner-wires was carried out by a solvent casting technique under aseptic conditions with and without incorporated antibiotics . Release kinetics of gentamicin and teicoplanin were studied in phosphate-buffered saline . Initial bacterial adhesion of Staphylococcus epidermidis on coated and bare implants was determined by radiolabelling and counts of detached viable organisms . RESULTS: The incorporated antibiotics showed a continuous release over a period of at least 96 h with an initial peak of release in the first 6 h . Attachment of non-viable microorganisms, detected by radiolabelled bacteria, was increased significantly by the polymer coatings (P < 0.05) . In contrast, the number of viable bacteria was reduced by the pure polymer (P < 0.01) and further by the polymer-antibiotic combinations (P < 0.05) . CONCLUSIONS: Poly(D,L-lactic acid) coating of implants could offer new perspectives in preventing biomaterial-associated infections . Combinations with other drugs to formulate custom-tailored implant surfaces are feasible. J Biotechnol, 2003 Mar 20, 101(3), 229 - 39 A modified Escherichia coli protein production strain expressing staphylococcal nuclease, capable of auto-hydrolysing host nucleic acid; Cooke GD et al.; The large-scale production of recombinant biotherapeutics, particularly recombinant proteins, provides significant process and regulatory challenges to the biotechnology industry in order to meet the regulatory agencies stringent requirements in a cost-effective manner . Host cell derived nucleic acid causes problems from both a process and a regulatory perspective, as high molecular weight chromosomal DNA is responsible both for the viscosity of cell lysates, and it is a source of heterologous DNA sequences whose inclusion in the final product must be prevented . We have constructed a modified Escherichia coli JM107 expression host (JMN), containing a staphylococcal nuclease expression cassette, integrated into the host chromosome at the dif locus . The nuclease is expressed as a fusion to the ompA signal peptide, and is translocated to the periplasm of the cell, protecting the cytoplasmic nucleic acid from any toxic activity . The nuclease is released during cell lysis, where it subsequently acts to hydrolyse host nucleic acid present in the lysate . Results with this strain show that sufficient levels of nuclease activity are produced to completely auto-hydrolyse the host's chromosomal DNA to a size non-visible on 1% agarose gel, generating a markedly lower lysate viscosity . This provides a suitable methodology to remove heterologous DNA sequences early in the product stream and decrease lysate viscosity, improving the efficiency of downstream processing and product yield, whilst avoiding the addition of exogenous nuclease and its prohibitive costs at large-scale. Biol Pharm Bull, 2003 Mar, 26(3), 365 - 7 Tryptanthrin inhibits interferon-gamma production by Peyer's patch lymphocytes derived from mice that had been orally administered staphylococcal enterotoxin; Takei Y et al.; Tryptanthrin, a biologically active compound found in the medicinal plant Polygonum tinctorium, reportedly has several biological activities . We investigated the effects of tryptanthrin on cytokine production by lymphocytes in response to staphylococcal enterotoxin B (SEB), which causes a variety of disorders in humans based on its induction of large amounts of immunostimulatory cytokines . Tryptanthrin dose-dependently inhibited interferon-gamma (IFN-gamma) and interleukin-2 production by mouse spleen cells and Peyer's patch (PP) lymphocytes in vitro . The efficacy of tryptanthrin was further studied in a mouse model in vivo . Tryptanthrin was administered orally 2 h after an oral challenge with SEB . Nineteen hours after SEB administration, PP lymphocytes were prepared, and IFN-gamma production by PP lymphocytes was examined . The production of IFN-gamma increased after SEB administration, and the elevated IFN-gamma production was significantly inhibited by tryptanthrin treatment . These results suggest that tryptanthrin may be effective in the treatment of disorders of the intestines, such as food poisoning, that are associated with activated lymphocytes. Eur J Dermatol, 2003 Jan-Feb, 13(1), 95 - 7 Adult purpura fulminans associated with staphylococcal infection and administration of colony-stimulating factors; Galimberti R et al.; Purpura fulminans (PF) is a rare syndrome of progressive haemorragic necrosis due to disseminated intravascular coagulation (DIC) and dermal vascular thrombosis leading to purpura and tissue necrosis . PF is more often associated with either a benign infection or a severe sepsis . Rarely, it has been related to drug intake . We report the case of a 24-year-old female patient who suffered from staphylococcal sepsis and pancytopenia, for which she was treated with antibiotics, granulocyte-colony stimulating factor (G-CSF) and granulocyte/macrophage CSF (GM-CSF) . Two days after the last GM-CSF dose, she developed widespread necrotic plaques with erythematous borders and purpura in the breast, arms and legs . Coagulation tests indicated DIC and a skin biopsy showed fibrin thrombi in the superficial dermal vessels . The patient totally recovered after removal of the necrotic tissues and application of skin autografts . Although staphylococcal infection was most probably involved in the development of PF, a role of CSF cannot be excluded in this case. Vox Sang, 2003 Feb, 84(2), 96 - 104 Inactivation of pathogens in platelet concentrates by using a two-step procedure; Mohr H et al.; BACKGROUND AND OBJECTIVES: Platelet concentrates are contaminated with residual leucocytes and may also be infected with viruses and bacteria . We investigated whether these pathogens can be inactivated by a two-step procedure comprising photodynamic treatment in the presence of the phenothiazine dye, thionine, followed by irradiation with ultraviolet light (UV-B, wavelength range 290-330 nm) . MATERIALS AND METHODS: Platelet concentrates were prepared from buffy coats . The concentrates were spiked with different viruses, bacteria and leucocytes, then illuminated with yellow light in the presence of thionine at dye concentrations between 1 and 5 microm and with UV-B at doses up to 2.4 J/cm2 . The infectivity of samples and the viability of leucocytes were assayed before and after treatment . The influence of treatment on in vitro platelet function was also examined . RESULTS: The inactivation of free viruses in platelet concentrates by photodynamic treatment with thionine/light was significantly enhanced when it was followed by irradiation with UV-B . The inactivation of leucocytes and of bacteria by UV-B was improved when it was preceded by thionine/light . Sterile platelet concentrates were prepared from buffy coats infected with Staphylococcus epidermidis . Platelet function and the storage stability of platelet concentrates were only moderately influenced by the two decontamination steps . CONCLUSIONS: Photodynamic treatment in the presence of the phenothiazine dye, thionine, followed by low-dose UVB, has the potential to inactivate viruses, leucocytes and bacteria, which might contaminate platelet concentrates . Both treatments complement each other. Ann Thorac Surg, 2003 Feb, 75(2), 593 - 6 Multiple pulmonary artery aneurysms due to infective endocarditis; Bozkurt AK et al.; Bilateral pulmonary artery aneurysms developed in the course of staphylococcal endocarditis in a 6-year-old girl with ventricular septal defect . Consecutive computed tomography scans revealed the progressive enlargement of one of the aneurysms . She underwent an urgent left upper lobectomy because of the impending rupture and a possible life-threatening hemorrhage . The second ipsilateral aneurysm was plicated in order to exclude the aneurysm sac . In the next operation the ventricular septal defect was closed and vegetations located on the tricuspid valve were removed . On follow-up spontaneous thrombotic resolution occurred in the right-sided aneurysms. Ann Thorac Surg, 2003 Feb, 75(2), 538 - 42 Treatment of recurrent staphylococcal mediastinitis: still a controversial issue; De Feo M et al.; BACKGROUND: Although surgical management with early debridement and closed mediastinal irrigation has proved successful in reducing early mortality following poststernotomy deep sternal wound infection, recurrence rates are still up to 20% . This study compared the effectiveness and safety of wound dressing with granulated sugar versus early muscle flap surgery in the management of recurrent postoperative Staphylococcal mediastinitis . METHODS: Between January 1995 and January 2002, 25 patients with severe recurrent staphylococcal mediastinitis were treated with granulated sugar wound dressing (group A) or with wound debridement, v-shape sternectomy and associated muscle flap surgery (group B) . Clinical outcomes and perioperative data were analyzed . Outcomes were compared between the groups evaluating the length of time for normalization of white blood cell (WBC) count and of body temperature and length of hospital stay . Patient characteristics determining best treatment option were identified . Survival and incidence of recurrence at follow-up were also analyzed . RESULTS: Study groups proved homogenous as to preoperative characteristics . Complete cure was achieved earlier in group A than in group B (defervescence: p = 0.0005; WBC normalization: p = 0.0001, respectively) . Hospital stay was shorter in group A . A statistically significant difference was found in hospital mortality (16% overall) between the two groups with better outcomes in group A (p = 0.039) . In the patient subset with the most severe preoperative profile (hemodialysis, tracheostomy, inotropic support) surgical treatment produced worse results than the sugar dressing method (p 0.048) . No case of recurrence was observed . CONCLUSIONS: Both treatments proved effective in recurrent type IV A Staphylococcal mediastinitis . Granulated sugar proved a safer option in severely compromised patients. Int J Impot Res, 2003 Feb, 15(1), 18 - 21 Antimicrobial activity of antibiotic-soaked, Resist-coated Bioflex; Hellstrom WJ et al.; This study investigates whether a hydrophilic coating (Resist), designed to inhibit bacterial adherence, applied to inflatable penile prostheses can prolong the effect of intraoperative antibiotics . The activity of antibiotic-soaked Bioflex (penile prosthetic substrate material) discs with and without Resist was examined by measuring the zone of inhibition following in vivo exposure in four groups of rabbits: 1, 2, 3 and 5 days' duration of disc implantation . Coated and uncoated discs were soaked in an aqueous solution of gentamicin and bacitracin . The implanted antibiotic-soaked discs were extracted, and the zone of inhibition against four microorganisms in vitro demonstrated that the Resist coating was especially effective against Staphylococcus epidermidis, and statistically significant improvements were observed for the coated over the uncoated substrate up to 3 days following implantation . This effect, and the anti-adherence properties of Resist, may prevent adhesion and colonization of some microorganisms to penile implants and reduce chances for infection. Proc Natl Acad Sci U S A, 2003 Mar 18, 100(6), 3191 - 6 Epub 2003 Feb 25. Structural basis for recognition by an in vitro evolved affibody; Hogbom M et al.; The broad binding repertoire of antibodies has permitted their use in a wide range of applications . However, some uses of antibodies are precluded due to limitations in the efficiency of antibody generation . In vitro evolved binding proteins, selected from combinatorial libraries generated around various alternative structural scaffolds, are promising alternatives to antibodies . We have solved the crystal structure of a complex of an all alpha-helical in vitro selected binding protein (affibody) bound to protein Z, an IgG Fc-binding domain derived from staphylococcal protein A . The structure of the complex reveals an extended and complementary binding surface with similar properties to protein-antibody interactions . The surface region of protein Z recognized by the affibody is strikingly similar to the one used for IgG(1) Fc binding, suggesting that this surface contains potential hot-spots for binding . The implications of the selected affibody binding-mode for its application as a universal binding protein are discussed. Plant Foods Hum Nutr, 2002 Fall, 57(3-4), 365 - 76 A preliminary study on the use of tempe-based formula as a weaning diet in Nigeria; Osundahunsi OF et al.; Tempe, an Indonesian mold fermented food, was prepared from cowpeas and soybeans using the traditional oriental process with modifications where appropriate . Four complementary foods were developed from whole maize meal or dehydrated fermented maize (ogi) flour fortified with either cowpea tempe or soybean tempe . Wholesomeness and potentials of the vanilla-flavored foods as weaning diets were determined . The nutrient content of all the developed products were within the range prescribed by the FAO/WHO pattern for processed weaning foods . The products had loose bulk densities between 0.40-0.55 g/ml and packed bulk densities between 0.68-0.75 g/ml; reconstitution indexes were between 92.30-104.00 g/ml; viscosities were 34.2-65.0 CP at 70 degrees C and 45.0-76.9 CP at 30 degrees C . Total plate counts ranged from 3.2 x 10(3) - 4.3 x 10(4) cfu/g; coliforms staphylococcus, mold, and yeast were absent in some of the products . The results on sensory attributes showed that tempe-based weaning diets were comparable with Mameal, a commercial product . Maize and maize-fortified diets were slightly higher in packed bulk density . Fortification reduced the viscosity of the diets and the values reduced with increasing temperature . Tempe-based weaning foods reconstituted easily in hot water, while cooking destroyed most of the microorganisms present . Maize-based tempe fortified foods were relatively inexpensive and have potential as weaning foods. Urol Res, 2003 Feb, 30(6), 394 - 8 Epub 2002 Dec 20. Granulocyte colony-stimulating factor for the treatment of biomaterial-associated staphylococcal infections in-vitro; Schlobe A et al.; Staphylococcal infections are a common and severe complication after the implantation of a prosthesis . We developed an in-vitro model for biomaterial-associated infections and studied the effects of human recombinant granulocyte colony-stimulating factor (rhuG-CSF; filgrastime) on the eradication of bacteria from the surface of biomaterial . Latex beads (25 micro m) were incubated with 10(7) colony forming units of either a slime producing (DSM 3269) or non-slime producing strain (ATCC 14990) of Staphylococcus epidermidis . Infected particles were consecutively confronted with effector cells, derived from heparinized whole blood samples taken from healthy volunteers, after stimulation with rhuG-CSF (5,000 IU/ml, 10,000 IU/ml) . Control blood specimens were not stimulated or conditioned with normal saline . The results indicate that stimulation with rhuG-CSF induced an increased rate of phagocytosis and lead to a more rapid reduction of adhering bacteria from the surface of the beads . Therefore, the in-vitro data suggest that patients with prosthesis infection may profit from an additional treatment with rhuG-CSF. Pediatr Surg Int, 2002 Dec, 18(8), 707 - 11 Epub 2002 Oct 24. Atypical mycobacterial disease in children: a personal series; Mushtaq I et al.; Atypical mycobacterial disease is common in children in Australia . Over 22 years, records were kept prospectively by the senior author . The diagnosis was confirmed in 118 patients, either by culture or by the combination of a positive skin test plus typical histology . There were 46 boys and 72 girls with a median age at diagnosis of 28 months . Most children (n = 56) presented with chronic lymphadenitis or abscess formation (n = 55) . The duration of illness varied from 4 days to 18 months . The most common sites affected were the head and neck (n = 112), with the pre-auricular region and anterior end of the submandibular triangle being characteristic . Nine patients had multifocal disease . The aim of treatment is to excise as much of the infected tissue as possible: 47 children had node excision through a planned incision that was closed primarily, with only 4 needing a second operation; 42 had excision of a node through the base of the superficial part of a collar-stud abscess with 6 recurrences . However, of the 33 children who had only drainage/curettage of the cavity or node 10 had recurrences requiring re-operation . Only 1 patient required a third operation . Morbidity was extremely low, with 1 staphylococcal wound infection . No child suffered permanent paresis of the mandibular division of the facial nerve . It is our belief that surgical excision of both the macroscopically affected and adjacent macroscopically unaffected nodes is necessary to achieve cure in the majority of cases. J Exp Bot, 2003 Mar, 54(384), 971 - 83 A Tudor protein with multiple SNc domains from pea seedlings: cellular localization, partial characterization, sequence analysis, and phylogenetic relationships; Abe S et al.; A major high molecular weight protein (HMP) in the cytoskeletal fraction from pea has been purified . A combination of chromatographic techniques and protease fragment analysis also facilitated the isolation of the encoding cDNA, disclosing the sequence of the complete open reading frame . The protein possesses four complete N-terminal Staphylococcal nuclease (SNc) domains, a central Tudor domain and a partial SNc domain at the C-terminus, which may act as a coiled-coil cytoskeleton interaction motif . Cell fractionation studies showed that the protein was abundant in the cytoskeleton fraction in dark-grown pea seedlings, but essentially was absent from the nucleus . Gel filtration column chromatography indicated that the native protein exists as a dimer, while isoelectric focusing suggested that there were at least four HMP isotypes . The protein co-eluted with ribosomes from a heparin affinity column in vitro, consistent with ribosome/polysome interactions in vivo . Significantly, sequence analysis of the C-terminal SNc motif may accurately predict nuclear versus cytoplasmic localization resulting in potentially very different functional roles for this protein family in different organisms . An antibody to HMP from peas was also raised and an HMP with a similar molecular mass was detected in the cytoskeleton fractions and to a lesser extent in the nuclear fraction (250 g pellet) from rice and wheat seedlings. J Am Vet Med Assoc, 2003 Feb 15, 222(4), 451 - 4 Isolation of Staphylococcus schleiferi from dogs with pyoderma; Frank LA et al.; OBJECTIVE: To determine frequency with which Staphylococcus schleiferi could be isolated from dogs with pyoderma and antimicrobial susceptibility patterns of isolates that were obtained . DESIGN: Prospective study . ANIMALS: 54 dogs with a first (n = 14) or recurrent (40) episode of pyoderma . PROCEDURE: Specimens were obtained and submitted for bacterial culture . Isolates were identified as S schleiferi on the basis of growth and biochemical characteristics . Two isolates were submitted for DNA sequencing to confirm identification . Methicillin susceptibility was determined by means of disk diffusion with oxacillin-impregnated disks . RESULTS: 3 of 14 dogs examined because of a first episode of pyoderma and 12 of 40 dogs examined because of a recurrent episode of pyoderma were receiving antimicrobials at the time of specimen collection . Staphylococcus schleiferi was not isolated from any dog with first-time pyoderma but was isolated from 5 dogs with recurrent pyoderma that were not receiving antimicrobials at the time of specimen collection and 10 dogs with recurrent pyoderma that were receiving antimicrobials . Nine isolates were identified as S schleiferi subsp schleiferi, and 6 were identified as S schleiferi subsp coagulans . All S schleiferi subsp schleiferi isolates were resistant to methicillin, but only 2 S schleiferi subsp coagulans isolates were . Two methicillin-resistant isolates were also resistant to fluoroquinolones, and 1 isolate had intermediate susceptibility to fluoroquinolones . CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that S schleiferi subsp schleiferi and S schleiferi subsp coagulans may be isolated from dogs with recurrent pyoderma . Although isolates from dogs with pyoderma were frequently resistant to methicillin, multiple drug resistance was uncommon. Surg Infect (Larchmt), 2001 Summer, 2(2), 171 - 5; discussion 175-7 New alternatives in management of the infected vascular prosthesis; Wilson SE; BACKGROUND: Infection of vascular prosthetics implanted for arterial occlusive disease occurs in approximately 1-5% of patients, including early and late clinical presentation . The incidence of infection depends on the anatomical site, with the highest rate occurring in vascular access grafts placed for hemodialysis and in inguinal and lower extremity incisions in patients undergoing bypass procedures for femoropopliteal tibial occlusion . Treatment of prosthetic graft infection has traditionally included antimicrobials, excision of the infected prosthesis, and extraanatomical bypass . With the recognition that prosthetic infection secondary to Staphylococcus epidermidis can be managed with less extensive procedures, clinical data have been reported on in situ replacement with antibiotic-bonded prostheses . Patients who have S . aureus isolated, including methicillin-resistant S . aureus, or gram-negative pathogens, still require conventional graft excision and extraanatomical bypass . METHODS: Selective review of the English-language literature . RESULTS: Recent clinical series show that management by graft excision of infected infrarenal aortic prosthetics and axillofemoral bypass results in 2- and 5-year survival rates of 67% and 47%, and limb salvage rates in survivors of 93% and 82% at 2 and 5 years . In situ replacement with a rifampicin-bonded prosthesis has been accomplished successfully in smaller numbers of patients and shows promising early results . Other methods under study include cryopreserved arterial and femoral vein allografts and autogenous femoral vein grafts, but data are limited when used as replacement for infected prosthetics . CONCLUSION: Advances in the management of infected vascular prostheses over the last decade have led to improved mortality and decreased amputation rates with conventional excision and extraanatomical bypass . Newer methods including in situ graft replacement with antibiotic-impregnated prosthetics appear suitable for low-virulence S . epidermidis infection . Early results are promising for cryopreserved allografts and autogenous femoral vein in situ grafts; however, more clinical experience and longer follow-up will be needed to confirm their durability in a contaminated field. Br J Surg, 2003 Mar, 90(3), 313 - 9 Serum antigen(s) drive the proinflammatory T cell response in acute pancreatitis; Sweeney KJ et al.; BACKGROUND: Immune cells and cytokines are central to the systemic inflammatory response syndrome and multiple organ failure associated with acute pancreatitis . The specific role of T cells in this response is unclear, and this study focused on evaluating T cell activation and its regulation in patients with acute pancreatitis . METHODS: Peripheral blood samples of 14 patients with acute pancreatitis were obtained within 24 h of the onset of pain, within 48 h and at 1 week . T cell expression of surface markers CD69, CD62L and CD25 was measured . The production of interleukin (IL) 10 and IL-2 in vitro in response to the superantigen Staphylococcus enterotoxin B (SEB) was assessed . Serum samples from these patients were co-cultured with peripheral blood mononuclear cells from volunteers in the presence or absence of cytotoxic T lymphocyte-associated antigen (CTLA) 4 immunoglobulin, a specific inhibitor of antigen-dependent T cell activation . RESULTS: Expression of CD69 was significantly increased in CD3(+) and CD4(+) populations at 48 h and 1 week, and on CD8(+) cells at 1 week . There was a significant increase in the production of SEB-induced IL-2 compared with findings in controls, but no significant IL-10 response . Serum from patients with pancreatitis activated normal T cells . This response was abolished completely by CTLA-4 . CONCLUSION: Acute pancreatitis results in the systemic activation of T cells . These cells are primed for a proinflammatory response to antigen stimulation and can be inhibited by antigen-specific T cell blockade . These data indicate that the immunoinflammatory response in acute pancreatitis is fueled by one or more serum antigens and offer prospects for further understanding of the aetiogenesis of pancreatitis . Proc Natl Acad Sci U S A, 2003 Mar 18, 100(6), 3185 - 90 Epub 2003 Feb 19. An affibody in complex with a target protein: structure and coupled folding; Wahlberg E et al.; Combinatorial protein engineering provides powerful means for functional selection of novel binding proteins . One class of engineered binding proteins, denoted affibodies, is based on the three-helix scaffold of the Z domain derived from staphylococcal protein A . The Z(SPA-1) affibody has been selected from a phage-displayed library as a binder to protein A . Z(SPA-1) also binds with micromolar affinity to its own ancestor, the Z domain . We have characterized the Z(SPA-1) affibody in its uncomplexed state and determined the solution structure of a Z:Z(SPA-1) protein-protein complex . Uncomplexed Z(SPA-1) behaves as an aggregation-prone molten globule, but folding occurs on binding, and the original (Z) three-helix bundle scaffold is fully formed in the complex . The structural basis for selection and strong binding is a large interaction interface with tight steric and polar/nonpolar complementarity that directly involves 10 of 13 mutated amino acid residues on Z(SPA-1) . We also note similarities in how the surface of the Z domain responds by induced fit to binding of Z(SPA-1) and Ig Fc, respectively, suggesting that the Z(SPA-1) affibody is capable of mimicking the morphology of the natural binding partner for the Z domain. Microbes Infect, 2003 Jan, 5(1), 13 - 7 Affinity of Helicobacter pylori to cholesterol and other steroids; Trampenau C et al.; Helicobacter pylori has a particular affinity to cholesterol . It is not known, however, whether other steroidal substances are bound as well . In order to characterize the specificity and nature of the H . pylori-steroid interaction, the affinity of H . pylori to cholesterol and several steroidal hormones was investigated . Seven strains of H . pylori (five reference strains, two wild strains) and one strain each of Staphylococcus epidermidis and Escherichia coli were cultured on a cholesterol-free medium . Cholesterol-free bacteria were incubated with cyclodextrin-mediated cholesterol and several cyclodextrin-mediated steroidal hormones (beta-estradiol, testosterone, progesterone, hydrocortisone, dexamethasone) . The steroid contents of the bacteria were determined by gas liquid chromatography . High amounts of cholesterol were detected in all H . pylori strains, whilst steroidal hormones were not found . Neither S . epidermidis nor E . coli showed an appreciable amount of cholesterol in the chromatographic examinations . Bacterial pretreatment with proteinase K diminished cholesterol adsorption of H . pylori . These data indicate a specific affinity of H . pylori to cholesterol . This unique property might serve as a pathogenicity component enabling survival and colonization of H . pylori in the gastric environment. Biomaterials, 2003 May, 24(10), 1829 - 31 Residual gentamicin-release from antibiotic-loaded polymethylmethacrylate beads after 5 years of implantation; Neut D et al.; In infected joint arthroplasty, high local levels of antibiotics are achieved through temporary implantation of non-biodegradable gentamicin-loaded polymethylmethacrylate beads . Despite their antibiotic release, these beads act as a biomaterial surface to which bacteria preferentially adhere, grow and potentially develop antibiotic resistance . In routine clinical practice, these beads are removed after 14 days, but for a variety of reasons, we were confronted with a patient in which these beads were left in situ for 5 years . Retrieval of gentamicin-loaded beads from this patient constituted an exceptional case to study the effects of long-term implantation on potentially colonizing microflora and gentamicin release . Gentamicin-release test revealed residual antibiotic release after being 5 years in situ and extensive microbiological sampling resulted in recovery of a gentamicin-resistant staphylococcal strain from the bead surface . This case emphasizes the importance of developing biodegradable antibiotic-loaded beads as an antibiotic delivery system . Surg Infect (Larchmt), 2001 Fall, 2(3), 225 - 9; discussion 229-30 Catheter materials affect the incidence of late blood-borne catheter infection; Mehall JR et al.; BACKGROUND: Adherence of bacteria and subsequent catheter-related infections (CRI) are greatly enhanced by the fibrin sheath that develops on indwelling catheters . Since the infection rate of catheters without fibrin sheaths is low and the fibrin sheath mediates bacterial adherence, catheter material is not thought to affect the incidence of late catheter-related infection . METHODS: A total of 276 ra