Staphylococcus

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 rats had catheters placed in the right jugular vein with the proximal end buried subcutaneously to eliminate exit site infection . Rats were divided into two groups: silastic catheters (SC; n = 133) and polyurethane catheters (PC; n = 143) . Injections of 1 x 10(8) CFU/mL of Staphylococcus epidermidis were given via the tail vein on either the day of surgery, day 0 (n = 53 SC, n = 51 PC), or on postoperative day 10 (n = 50 SC, n = 62 PC) . Thirty animals from each group (SC, PC) received sterile saline injections on day 10 and served as controls . Animals were sacrificed on postinjection day 3 . Catheters were removed via the chest and placed into trypticase soy broth . Broth was incubated at 37 degrees C for 48 h . Microscopy for the fibrin sheath was done on 20 randomly selected catheters (10/group) . Data were compared using Fisher's exact test, with p < 0.05 considered significant . RESULTS: Incidence of CRI was equal prior to the formation of the fibrin sheath, while CRI was significantly higher in silastic catheters in the presence of a fibrin sheath . Without a fibrin sheath (day 0), 8/53 silastic catheters and 3/51 polyurethane catheters were infected (p = NS) . With a fibrin sheath (day 10), 31/50 silastic catheters were infected versus 20/62 polyurethane catheters (p < 0.05) . Control catheters were all culture negative (30/group) . With light microscopy, 20/20 catheters had fibrin sheaths at day 10 with no visible difference between silastic and polyurethane catheters . CONCLUSION: Catheter material does affect the incidence of catheter-related infection even when catheters are coated with a fibrin sheath . This difference may relate to a difference in the fibrin sheath itself as it forms on different catheter materials.

Surg Infect (Larchmt), 2002 Spring, 3(1), 11 - 9
Outcome analysis of intraabdominal infection with resistant gram-positive organisms; Pelletier SJ et al.; BACKGROUND: Although the microbiology of intraabdominal infection has been well described, the role of resistant organisms remains unclear . To evaluate the hypothesis that intraabdominal infections from resistant gram-positive cocci (rGPC) have worse outcomes compared to those with susceptible organisms, patient characteristics and outcomes were compared between these groups . METHODS: Analysis of peritoneal infections was performed on prospectively collected data of all consecutive surgical infections from December 1996 to June 1999 at a university hospital . Intraabdominal infection was defined either by a positive peritoneal cavity culture or on clinical grounds (e.g., abscess), which prompted antimicrobial or surgical therapy . Resistant Staphylococcus and Enterococcus spp . were defined as those strains resistant to oxacillin, gentamicin, or vancomycin . RESULTS: Compared to episodes of intraabdominal infection from susceptible organisms (n = 365), infections due to rGPC (n = 52) were associated with an increased severity of illness (p < 0.0001), longer time from admission to treatment (p < 0.0001), longer duration of therapy (p = 0.008), greater proportion of nosocomial infection (p < 0.0001), increased length of stay (p < 0.0001), and an increased mortality rate (9% versus 23%; p = 0.003) . However, comparison of intraabdominal infection with rGPC to a group controlled for severity of illness demonstrated a prolonged time from admission until treatment and longer duration of hospitalization but a similar mortality rate between groups (17% versus 23%; p = 0.46) . CONCLUSION: Intraabdominal infection with rGPC is an indicator of poor prognosis and severe illness . Although not an independent predictor of mortality, the significantly increased duration of therapy and prolonged duration of hospitalization may have considerable economic impact.

J Pediatr Surg, 2003 Jan, 38(1), 138 - 42; discussion 138-42
Surgical infants on total parenteral nutrition have impaired cytokine responses to microbial challenge; Cruccetti A et al.; BACKGROUND/PURPOSE: Cytokines are essential for the prevention of microbial infections . Total parenteral nutrition (TPN) in infancy is associated with an increased risk of infection, and this could be related to altered cytokine production . The aim of the study was to determine if cytokine production is altered in monocytes from surgical infants receiving TPN . METHODS: There were 3 study groups: (a) infants receiving TPN, (b) enterally fed healthy control infants, and (c) enterally fed healthy control adults . Blood samples were incubated with either Escherichia coli LPS, Staphylococcus epidermidis, or with medium alone . Flow cytometry was used to measure monocyte intracellular cytokine: tumor necrosis factor (TNF)-alpha, interleukin (IL)-6, IL-1beta . RESULTS: After LPS stimulation, the percent of monocytes producing TNF-alpha and IL-6 were lower in infants on TPN than both control infants and adults . This was most apparent for TNF-alpha . The difference for IL-1beta was significant only between infant on TPN and control adults . When blood was stimulated with S . epidermidis, all 3 cytokines were significantly lower in the TPN group compared with control adults . However, the differences between infants on TPN and infant controls only reached statistical significance for IL-6 . CONCLUSIONS: The inflammatory response to bacterial challenge is impaired in infants on TPN compared with enterally fed infants or adults . The pattern of this response may be dependent on the nature of the microbial challenge . Our results indicate that the susceptibility of TPN-fed surgical infants to bacterial infections may in part be caused by impaired cytokine responses after bacterial invasion .

Vet Parasitol, 2003 Mar 10, 112(3), 241 - 7
Efficacy of selamectin administered topically in the treatment of feline otoacariosis; Blot C et al.; The efficacy of a novel avermectin, selamectin (Stronghold, Pfizer), was evaluated against naturally acquired aural infestations of Otodectes cynotis . Selamectin was administered topically in a single spot to the skin of each animal's back at the base of the neck in front of the scapulae at a minimum dosage of 6mgkg(-1) . Thirty cats of a cattery of 120 animals with an endemic infestation were treated on days 0 and 30 . Including untreated control cats was therefore not possible because of animal welfare consideration . Clinical examination and visualization of mites by otoscopic examination of the external ear canal and microscopic examination of aural debris/exudate were performed twice weekly from days 0 to 30 . On day 0 (and day 28 if the ears canals were erythematous), a swab was taken from each canal and sent to bacteriology and mycology units for detection and identification of bacteria (Staphylococcus mainly) and yeast (Malassezia pachydermatis only).Selamectin was safe, parasites were killed before day 3 and eliminated before day 17 . Selamectin was 100% effective against natural aural infestations, even if erythema (26-33% of cats) and scratch reflex (23-40% of cats) persisted for 2 weeks after the cats tested negative for mites.

Biochemistry, 2003 Feb 25, 42(7), 2056 - 61
Carboxylate groups on the manganese-stabilizing protein are required for efficient binding of the 24 kDa extrinsic protein to photosystem II; Bricker TM et al.; The effects of the modification of carboxylate groups on the manganese-stabilizing protein on the binding of the 24 kDa extrinsic protein to Photosystem II were investigated . Carboxylate groups on the manganese-stabilizing protein were modified with glycine methyl ester in a reaction facilitated by 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide . The manganese-stabilizing protein which was modified while associated with NaCl-washed membranes could bind to calcium chloride-washed PS II membranes and reconstitute oxygen evolution in a manner similar to that observed for unmodified manganese-stabilizing protein (Frankel, L.K, Cruz, J . C . and Bricker, T . M . (1999) Biochemistry 38, 14271-14278) . However, PS II membranes reconstituted with this modified protein were defective in their ability to bind the extrinsic 24 kDa protein of Photosystem II . Mapping of the sites of modification was carried out by trypsin and Staphylococcus V8 protease digestion of the modified protein and analysis by MALDI mass spectrometry . These studies indicated that the domains (1)E-(71)D, (97)D-(144)D, and (180)D-(187)E are labeled when the manganese-stabilizing protein is bound to NaCl-washed Photosystem II membranes . We hypothesize that modified carboxylates, possibly residues (1)E, (32)E, (139)E, and/or (187)E, in these domains are responsible for the altered binding affinity of the 24 kDa protein observed.

Am J Med Sci, 2003 Feb, 325(2), 101 - 5
Churg-Strauss syndrome: diagnostic difficulties and pathogenesis; Diri E et al.; A 52-year-old woman with a history of chronic obstructive pulmonary disease presented with symmetrical polyarthritis involving her metacarpophalangeal and proximal interphalangeal joints, knees, ankles, and hips and with a purpuric rash involving her lower extremities . She had a history of recurrent episodes of purulent otitis often associated with myalgias and arthralgias . Laboratory studies at presentation included leukocytosis with 16% eosinophils, an elevated rheumatoid factor titer, and an elevated antineutrophil cytoplasmic antibody titer . Cultures from the right ear canal grew . Skin biopsy revealed leukocytoclastic vasculitis with pericapillary eosinophils . The patient was treated with prednisone and then with azathioprine after the rash relapsed during the tapering of prednisone . Four months after her initial presentation, she developed bilateral foot drop . A sural nerve biopsy revealed vasa nervosum vasculitis . The diagnosis of Churg-Strauss syndrome was established, and she was treated with an increased dose of azathioprine and a slowly tapering prednisone regimen . This case report suggests that patients with Churg-Strauss syndrome can present with a syndrome suggesting rheumatoid arthritis . In this particular patient, recurrent staphylococcal infections may have triggered the vasculitic process.

Clin Experiment Ophthalmol, 2003 Feb, 31(1), 40 - 3
Penetration of oral and topical ciprofloxacin into the aqueous humour; Ghazi-Nouri SM et al.; AIMS: To investigate the penetration of ciprofloxacin via different modes of administration into the aqueous humour using capillary zone electrophoresis and to determine its prophylactic role in ophthalmic surgery . METHODS: A prospective study was conducted of 84 patients, divided into two groups, undergoing routine cataract surgery and intraocular lens implantation . Forty patients received 750 mg ciprofloxacin orally the evening before and on the morning of surgery 12 h apart . Forty-four patients received one drop of 0.3% topical ciprofloxacin 90 and 30 min prior to surgery . At the commencement of surgery 0.1-0.2 mL of aqueous fluid was sampled from the anterior chamber and assayed for ciprofloxacin concentration using capillary-zone electrophoresis . RESULTS: The concentration of ciprofloxacin in the group receiving topical doses was less than 0.1 micro g/mL, which is well below the minimum inhibitory concentration for cipro-floxacin inhibiting 90% (MIC90) of strains of Staphylococcus epidermidis (0.4 micro g/mL) . The mean concentration of ciprofloxacin in the aqueous humour of the oral group was 0.26 micro g/mL with a standard deviation of 0.12 micro g/mL (range 0.09-0.67 micro g/mL) with only 12.5% achieving levels higher than MIC90 for S . epidermidis . CONCLUSION: The topical ciprofloxacin 0.3% as given in this study achieved poor aqueous humour concentration . Oral ciprofloxacin given in this regime should not on its own be considered adequate prophylaxis in intraocular surgery . Capillary zone electrophoresis can be used as an alternative tool to the existing high-pressure liquid chromatography methods for analysing ciprofloxacin level in the aqueous humour.

J Am Chem Soc, 2003 Feb 19, 125(7), 1748 - 58
Side chain dynamics in unfolded protein states: an NMR based 2H spin relaxation study of delta131delta; Choy WY et al.; NMR relaxation data on disordered proteins can provide insight into both structural and dynamic properties of these molecules . Because of chemical shift degeneracy in correlation spectra, detailed site-specific analyses of side chain dynamics have not been possible . Here, we present new experiments for the measurement of side chain dynamics in methyl-containing residues in unfolded protein states . The pulse schemes are similar to recently proposed methods for measuring deuterium spin relaxation rates in (13)CH(2)D methyl groups in folded proteins.(1) However, because resolution in (1)H-(13)C correlation maps of unfolded proteins is limiting, relaxation data are recorded as a series of (1)H-(15)N spectra . The methodology is illustrated with an application to the study of side chain dynamics in delta131delta, a large disordered fragment of staphylococcal nuclease containing residues 1-3 and 13-140 of the wide-type protein . A good correlation between the order parameters of the symmetry axes of the methyl groups and the backbone (1)H-(15)N bond vectors of the same residue is observed . Simulations establish that such a correlation is only possible if the unfolded state is comprised of an ensemble of structures which are not equiprobable . A motional model, which combines wobbling-in-a-cone and Gaussian axial fluctuations, is proposed to estimate chi(1) torsion angle fluctuations, sigma(chi)()1, of Val and Thr residues on the basis of the backbone and side chain order parameters . Values of sigma(chi)()1 are approximately 10 degrees larger than what has previously been observed in folded proteins . Of interest, the value of sigma(chi)()1 for Val 104 is considerably smaller than for other Val or Thr residues, suggesting that it may be part of a hydrophobic cluster . Notably large (15)N transverse relaxation rates are observed in this region . To our knowledge, this is the first time that side chain dynamics in an unfolded state have been studied in detail by NMR.

Yan Ke Xue Bao, 1999 Dec, 15(4), 267 - 9
{Study on bacterial strains in conjunctival sac before and after intraocular surgery}; Yang W et al.; PURPOSE: To investigate the changes of bacterial strains in the conjunctival sac before and after intraocular surgery, as well as clinical effects of preoperative antibiotic agents . METHODS: One hundred and thirty-three eyes with intraocular surgery included 68 eyes with cataract surgery, 39 eyes with glaucoma surgery, 20 eyes with retinal detachment surgery and 6 eyes with vitretomy . Bacterial culture and pharmacologic sensitivity test were performed on the conjunctival smear from the 133 eyes with intraocular surgery before and after the operation and the anterior chamber aspirate from the 68 eyes with cataract surgery after the operation . RESULTS: At the time of admission, 61 eyes(45.8%) of 133 eyes with intraocular surgery were culture-positive conjunctival smears with the following identified: Staphlococcus aureus: 23 eyes; Staphlococcus epidermidis: 25 eyes; Gram-positive rods: 13 eyes . 8 eyes(6%) had culture-positive conjunctival smears before disinfection, (Staphlococcus aureus: 3 eyes; Staphlococcus epidermidis: 5 eyes) . One eye(1.5%) had culture-positive Staphylococcus epidermidis in anterior chamber aspirate of 68 eyes with cataract surgery at conclusion of surgery . Also, 4 eyes(3%) were culture-positive conjunctival smears after surgery . All the bacterial strains of the group were sensitive to Tobramycin, Gentamycin, Cephradinum(Velosel), Vancomycinum and Rifampin, but they were resistance to Tetracyclinum, Chloromycetin, Erythromycin and Ofloxacin . Eye Science 1999; 15: 267-269 . CONCLUSION: In order to prevent bacterial endophthalmitis after intraocular surgery, installation of an useful antibiotic eye drop before the surgery is important.

Arthroscopy, 2003 Feb, 19(2), 172 - 81
Surgical site infections after arthroscopy: Outbreak investigation and case control study; Babcock HM et al.; PURPOSE: The goal of this study was to determine the causes of increased post-arthroscopy surgical site infections (SSIs) and to define risk factors for infection . Type of Study: Outbreak investigation and case control study at a university-affiliated community hospital from 1994 to 1996, with surveillance through 1999 . METHODS: Demographic, clinical, and microbiological data were collected on 27 post-arthroscopy SSIs from 1994 through 1999 . Risk factors for SSI were identified by case-control analysis and presented as odds ratios (OR) and 95% confidence intervals (CI) . RESULTS: Initial investigation revealed an increased annual rate of post-arthroscopy SSIs in 1995 (1.3%) . Infection control deficiencies were identified, and feedback was provided to surgeons and staff . Instrument sterilization was standardized, flash sterilization prohibited, and preoperative shaving discouraged . Case-control analysis of 10 cases (from 1994 to 1996) found a statistically significant increase in risk of SSI with intra-articular corticosteroid joint injection (OR, 9.33; 95% CI, 1.6 to 64.9); other risk factors did not reach statistical significance . SSI rates dropped after feedback and education (0.34% in 1996) . Continued surveillance revealed 2 smaller outbreaks, in December 1997 (1997 rate, 1.13%) and September 1998 (1998 rate, 1.09%) . Case-control analysis of the 17 cases occurring in 1997 through 1999 was also performed . The 1997 outbreak appeared to be related to preoperative razor shaving (P =.003), which was then prohibited by hospital policy . One scrub nurse was also associated with 75% of these cases, which were culture-positive for coagulase-negative Staphylococcus . The cases in the 1998 outbreak shared prolonged procedure duration and conversion to arthrotomy . Of 27 cases, 24 required repeat hospitalization and repeat surgery, at an average excess cost of $9,154.84 per case . All received prolonged courses of intravenous or oral antibiotics . CONCLUSIONS: Post-arthroscopy SSIs are associated with significant morbidity and cost . Although small numbers make finding statistical significance difficult in case-control studies, infection control and CDC-recommended interventions can lower SSI rates . Careful definitions, ongoing surveillance, and long-term follow-up are helpful in reporting results of infection control interventions.

Clin Orthop, 2003 Jan, (406), 246 - 52
Bacterial skin contamination after surgical preparation in foot and ankle surgery; Ostrander RV et al.; An effective presurgical preparation is an important step in limiting surgical wound contamination and preventing infection . The purpose of this study was to evaluate residual bacterial skin contamination after surgical skin preparation in foot and ankle surgery to determine if current techniques are satisfactory in eliminating harmful pathogens . Fifty consecutive patients having surgical procedures of the foot and ankle were studied . Each lower extremity was prepared randomly with either a one-step povidone-iodine topical gel or a two-step iodophor scrub followed by a povidone-iodine paint . After preparation and draping, cultures were obtained at three locations: the hallux nailfold, web space between the second and third, and fourth and fifth toes, and the anterior ankle (control) . In the gel group, positive cultures were obtained from 76% of halluces, 68% of toes, and 16% of controls . In the scrub and paint group, positive cultures were obtained from 84% of halluces, 76% of toes, and 28% of controls . Numerous pathogens were cultured, with Staphylococcus epidermidis being the most prevalent . Based on the findings of the current study, presurgical skin preparation with a povidone-iodine based topical bactericidal agent is not sufficient in eliminating pathogens in foot and ankle surgery . The unique environment of the foot and its resident organisms may play a role in the higher infection rates associated with surgery of the foot and ankle.

Water Sci Technol, 2003, 47(1), 303 - 8
Isolation of naphthalene-degrading bacteria from tropical marine sediments; Zhuang WQ et al.; Oil pollution is a major environmental concern in many countries, and this has led to a concerted effort in studying the feasibility of using oil-degrading bacteria for bioremediation . Although many oil-degrading bacteria have been isolated from different environments, environmental conditions can impose a selection pressure on the types of bacteria that can reside in a particular environment . This study reports the successful isolation of two indigenous naphthalene-degrading bacteria from oil-contaminated tropical marine sediments by enrichment culture . Strains MN-005 and MN-006 were characterized using an extensive range of biochemical tests . The 16S ribosomal deoxyribonucleic acid (rDNA) sequence analysis was also performed for the two strains . Their naphthalene degradation capabilities were determined using gas chromatography and DAPI counting of bacterial cells . Strains MN-005 and MN-006 are phenotypically and phylogenetically different from each other, and belong to the genera Staphylococcus and Micrococcus, respectively . Strains MN-005 and MN-006 had maximal specific growth rates (micro(max)) of 0.082 +/- 0.008 and 0.30 +/- 0.02 per hour, respectively, and half-saturation constants (K(s)) of 0.79 +/- 0.10 and 2.52 +/- 0.32 mg per litre, respectively . These physiological and growth studies are useful in assessing the potential of these indigenous isolates for in situ or ex situ naphthalene pollutant bioremediation in tropical marine environments.

Shock, 2003 Feb, 19(2), 163 - 8
Trypanosoma cruzi sensitizes mice to fulminant SEB-induced shock: overrelease of inflammatory cytokines and independence of Chagas' disease or TCR Vbeta-usage; Paiva CN et al.; Trypanosoma cruzi-infected mice display increased susceptibility to shock induced by injection of lipopolysaccharide (LPS), anti-CD3, or resulting from interleukin (IL)-10-defective response to the parasite itself, but the basis of such susceptibility remains unknown . Herein, we tested the susceptibility of mice inoculated with virulent and avirulent T . cruzi to staphylococcal enterotoxins (SE), potent inducers of inflammatory cytokine secretion . Mice infected with T . cruzi CL-strain or inoculated with the avirulent clone CL-14, a clone that does not induce disease or polyclonal lymphocyte activation, succumb suddenly to low doses of staphylococcal enterotoxin B (SEB), but not to staphylococcal enterotoxin A (SEA) . High plasma levels of TNF, IFN-gamma, and liver transaminases alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were found in these mice, indicating lethal toxic shock . Sensitization to shock required inoculation of live avirulent trypomastigotes and a time interval before challenge with SEB . We found no prior skewing of T cell receptor (TCR) Vbeta-repertoire in CL-14-inoculated mice that could be responsible for sensitization . Splenocytes from CL-14-inoculated mice proliferated more under anti-Vbeta8 than anti-TCRbeta stimulation when compared with normal mice, but were suppressed to SEB stimulation . Both SEB and anti-Vbeta8 antibodies stimulated splenocytes from T . cruzi-inoculated mice to secrete higher levels of inflammatory cytokines than normal controls . Taken together, our results show that T . cruzi inoculation can sensitize mice to lethal SEB-induced shock even in the absence of tissue damage, polyclonal lymphocyte activation, or previously increased levels of inflammatory cytokines, and they suggest that altered reactivity of Vbeta8 lymphocytes may be involved in the phenomenon.

Chest, 2003 Feb, 123(2), 488 - 98
Pulmonary infiltrates in patients receiving long-term glucocorticoid treatment: etiology, prognostic factors, and associated inflammatory response; Agusti C et al.; BACKGROUND: Glucocorticoid treatment alters immunoregulatory defense mechanisms and may therefore favor the development of different pulmonary infections . METHODS: The etiology, prognostic factors, and associated inflammatory response of pulmonary infiltrates in 33 patients receiving long-term glucocorticoid treatment (LTGCT) were prospectively evaluated . RESULTS: Aspergillus spp (n = 9, 31%) and Staphylococcus spp (n = 6, 21%) were the most common causative agents . Using different diagnostic techniques, we obtained a specific diagnosis in 28 of 33 episodes (85%) of pulmonary infiltrates . Bronchoscopic techniques provided the diagnosis in 64% of the cases . Crude mortality was 45% . Variables associated with mortality were as follows: age > 64 years, bilateral radiographic involvement, delay in diagnosis, inappropriate empirical treatment, Simplified Acute Physiology Score (SAPS) II > or = 25, and requirement for mechanical ventilation (MV) . SAPS II > or = 25 (odds ratio {OR}, 16; 95% confidence interval, 1 to 260) and MV requirement (OR, 50; 95% confidence interval, 2 to 360) were also significant on multivariate analysis . Pulmonary infections were associated with an increase in the concentration of relevant inflammatory cytokines such as tumor necrosis factor-alpha and interleukin-6 both in serum and BAL . This local and systemic inflammatory response was attenuated when compared with the response observed in patients with pulmonary infections but without glucocorticoid treatment or receiving glucocorticoids for a short period of time (< 9 days) . CONCLUSIONS: Pulmonary infiltrates in patients receiving LTGCT are often caused by fungi and Gram-positive cocci, and are associated with attenuated local and systemic inflammatory response . Although in most cases, sputum cultures and bronchoscopic techniques are diagnostic, the associated mortality is high, particularly in those requiring MV.

Cell Immunol, 2002 Oct, 219(2), 98 - 107
Activation-induced cell death in human T cells is a suicidal process regulated by cell density but superantigen induces T cell fratricide; Gorak-Stolinska P et al.; Repeated ligation of the TCR results in apoptosis (activation-induced cell death; AICD) . Superantigens such as Staphylococcal enterotoxin B (SEB) are particularly efficient at inducing AICD in T cells . We investigated whether apoptosis in human T cell subsets was due to fratricide (killing of neighboring cells) or suicide (cell autonomous death) . AICD of Th1, Th2, Tc1, and Tc2 effector cells was dramatically enhanced at low cell densities and could be observed in single cell microcultures . AICD was unaffected by adhesion molecules or neighboring cells undergoing AICD, confirming the predominance of a suicidal mechanism . However, SEB was able to induce fratricidal apoptosis of type 1, but not type 2 cells . Fratricide was also observed when unstimulated T cells were exposed to activated Tc1 effector cells . Thus, AICD is tightly regulated to allow clonal T cell expansion and memory cell generation, but superantigens may subvert this process by allowing T cell fratricide.

Hunan Yi Ke Da Xue Xue Bao, 2002 Feb 28, 27(1), 21 - 2
{Detection of staphylococcal clumping factor with purified fibrinogen}; Chen J et al.; A modified method of detecting staphylococcal clumping factor with purified fibrinogen was introduced . The results showed that the method was simple and concordant with the one with rabbit plasma . Moreover, the results were easy to observe . The traditional method with human plasma was found to be likely to produce false positive.

Hua Xi Yi Ke Da Xue Xue Bao, 2002 Apr, 33(2), 299 - 301, 304
{Study on preparation and tissue distribution of SEA liposomes}; Li Z et al.; OBJECTIVE: To make a study on the preparation and tissue distribution of staphylococcal enterotoxin A (SEA) liposomes and to provide scientific basis for the therapy of liver cancer by using SEA liposomes . METHODS: SEA liposomes were prepared by reverse-phase evaporation; the diameter and entrapment efficiency (EC) of SEA liposomes were determined . 125I labeled SEA solution and 125I-SEA liposomes were administrated intravenously to mice, respectively . The radioactivity of the organs was determined by gamma-counter . RESULTS: The mean diameter and EC of SEA liposomes were 505 +/- 34 nm and 44.1% +/- 4.8%, respectively . SEA liposomes were found mainly distributed in the liver and spleen . SEA liposomes had a higher blood clearance, compared with SEA solution; SEA solution had high-radio-activity in plasma and kidney; there was statistical significance between the two groups (P < 0.05) . CONCLUSION: The preparation method of SEA liposomes is simple and repeatable . SEA liposomes possess liver-targeting properties and may provide a new application foreground for the treatment of liver cancer.

Zhong Yao Cai, 2000 May, 23(5), 272 - 4
{Study on the inhibitory activity, in vitro, of baicalein and baicalin against skin fungi and bacteria}; Yang D et al.; In this paper, we concentrated in examining, in vitro, the antiseptic activity of the baicalein and baicalin upon the seventeen pathogenic skin fungal and sixteen skin bacterial strains, these two flavonic compounds were known principally as the biosubstances of a traditional Chinese medicinal plant: Scutellaria baicalensis . In agar media, the baicalein possessed potent specific activity against the pathogenic yeasts with MICs of 70-100 micrograms/ml; But in the same condition, no inhibitory effect was observed upon dermatophytes and filamentous imperfect fungi for baicalein, and upon all used strains for baicalin . According to the antibacterial test of baicalein, a high efficacy was achieved against certain causative specie of axillary and foot's odour such as Micrococcus sedentarius, Staphylococcus epidermidis, S . hominis and C . xerosis with a MICs inferior to 250 micrograms/ml . The good inhibitory activity of baicalein could be linked to the group hydroxyl (-OH) in position seven of the molecule.

Neth J Med, 2002 Oct, 60(9), 349 - 53
Th1/Th2 cytokine imbalance in a family with hyper-IgE syndrome; Netea MG et al.; BACKGROUND: Hyperimmunoglobulin E (hyper-IgE) syndrome is a rare immunodeficiency characterised by recurrent skin and respiratory tract infections, skeletal and dental abnormalities, chronic eczema, and elevated serum IgE . We describe a family with four hyper-IgE syndrome patients (38, 37, 30 and 7 years old), in which we investigated the cytokine response to both specific and non-specific stimulation . METHODS: Whole blood from patients and volunteers was stimulated for either 24 or 48h at 37 degrees C with heat-killed Staphylococcus, C . albicans or a combination of IL-12 and IL-18 . Cytokine concentrations in the plasma were measured by specific radioimmuno-assays or ELISA . RESULTS: Serum IgE ranged from 5,000 to 16,670 IU/ml, and neutrophil chemotaxis was normal in all four patients . Tumour necrosis factor, interleukin (IL)-1beta, IL-6 and IL-8 production after stimulation of whole-blood cultures with lipopolysaccharide or heat-killed S . aureus did not differ between the adult patients and four healthy controls . In contrast, when blood from patients and controls was stimulated with heat-killed S . aureus or C . albicans, a severe imbalance towards a Th2 phenotype was found, with 10- to 30-fold reduction in the IFNgamma/IL-10 ratios in the hyper-IgE syndrome patients . The IFNgamma production in the patients was less severely impaired when blood was non-specifically stimulated with a combination of IL-18 and IL-12 . CONCLUSION: In this family with hyper-IgE syndrome, the imbalance in the Th1/Th2 cytokine production may have been involved in the pathogenesis of the recurrent infections and/or chronic eczema characteristic of this disease.

Clin Exp Allergy, 2002 Nov, 32(11), 1552 - 7
Polyclonal and allergen-induced cytokine responses in children with elevated immunoglobulin E but no atopic disease; Smart JM et al.; BACKGROUND: Reduced Th1 and elevated Th2 cytokine responses are considered to be a principal mechanism in the generation of the inflammation leading to the manifestations of atopic disease in the skin of atopic dermatitis and in the airways of asthma . If reduced Th1 and elevated Th2 responses are principal determinants of the manifestation of atopic disease it might be expected that subjects with established disease would exhibit differences in their cytokine profiles as compared with atopic patients without clinical disease . OBJECTIVE: To determine whether asymptomatic atopic children exhibit a cytokine imbalance similar to that seen in patients with established atopic disease or if they behave like non-atopic controls . Cytokine responses in a group of children with elevated IgE but no clinical manifestations of disease, atopic children with established disease and non-atopic controls were compared . METHODS: We examined allergen-induced (house dust mite, HDM, rye grass pollen and RYE) cytokine responses in parallel with polyclonal (staphylococcal enterotoxin B, SEB) cytokine responses in a group of children with elevated serum IgE levels without current or past evidence of atopic disease (median age 6.6 years) and compared these with a non-atopic control group (median age 6.5 years) and a group of children with atopic disease (median age 6.7 years) . RESULTS: Symptomatic atopic children had reduced SEB-induced IFN-gamma and increased SEB-induced IL-4 and IL-5 as compared with non-atopic controls . In contrast, SEB-induced IFN-gamma, IL-4 and IL-5 production in asymptomatic atopics was not significantly different from the non-atopic control subjects . Allergen-induced Th1 (IFN-gamma) and Th2 (IL-5 and IL-13) cytokine production was increased in both symptomatic atopics and asymptomatic atopics when compared with non-atopic controls . CONCLUSION: The defect in polyclonally induced IFN-gamma production was associated with the clinical manifestation of atopic disease but not the atopic stateper se . This suggests that the global reduction in IFN-gamma is the key determinant of the development of overt atopic disease . In contrast, elevated allergen-induced Th2 cytokine responses in children related to the atopic state per se irrespective of the presence of clinical atopic disease.

Sheng Wu Gong Cheng Xue Bao, 2002 Sep, 18(5), 597 - 600
{Cloning of staphylococcal enterotoxin B gene and its highly expression in Escherichia coli}; Yang LQ et al.; An about 700 bp DNA fragment was amplified from genome DNA of S . aureus TSTw by PCR . This fragment was cloned into pGEM-7Zf(+) and the recombinant plasmid was transformed into E . coli DH5 alpha . The sequencing result of the recombinant plasmid demonstrated that it contains seb gene with 717 bp (without signal encoding region of 81 bp) which has the same nucleotide sequence as described in literature . The seb gene was cloned into expression vector 7ZTS and was transformed into E . coli JM109 (DE3) . The expression level of SEB was as high as 33.3% of the cell total proteins.

Proteins, 2003 Feb 15, 50(3), 507 - 15
Molecular dynamics simulation reveals a surface salt bridge forming a kinetic trap in unfolding of truncated Staphylococcal nuclease; Gruia AD et al.; Surface salt bridges are ubiquitous in globular proteins . Their contribution to protein stability has been extensively debated in the past decade . Here, molecular dynamics simulations are performed starting from a non-equilibrium state of Staphylococcal nuclease (SNase) with C-terminal truncation (SNaseDelta) . The results indicate a key role in the unfolding of the surface salt bridge between arginine 105 and glutamate 135 . Experimentally, SNaseDelta is known to be partially unfolded . However, in simulations over 1 ns at 300 K and over 500 ps at 400 K, SNaseDelta remains stable in the native-like folded conformation, the salt bridge hindering unfolding . When the potential function is altered so as to selectively weaken the salt bridge, which then breaks rapidly at 430 K, the protein starts to unfold . The results suggest that breaking of this salt bridge presents a significant barrier to the unfolding transition of SNaseDelta from a native-like state to the unfolded state . Potential of mean force calculations indicate that the barrier height for this transition is approximately 7 kcal/mol .

Ann Cardiol Angeiol (Paris), 2000 Sep, 49(6), 347 - 50
{Staphylococcus epidermidis infective endocarditis after mitral surgery, successfully treated with aspirin and antibiotics: a case report}; Meimoun P et al.; We report a case of infectious endocarditis from Staphylococcus epidermidis that occurred early after mitral valve repair (one month), suggested by fever of 38.5 degrees C and valvular vegetations of less than 10 mm on the mitral valve . In the absence of standard recommendations (medical or surgical) in treating patients with infectious endocarditis occurring after mitral valve repair, and in the absence of complications, a medical regimen was chosen associating aspirin (anti-aggregant dosages) with antibiotics . The benefit of aspirin in endocarditis has been demonstrated in experimental studies with regards to valvular vegetations and embolic risk but remains to be studied in human clinical trials . With the association of aspirin (100 mg/d) and triple antibiotic therapy (rifampicin 1200 mg/d, vancomycin 2 g/d, gentamycin 180 mg/d), the clinical status improved with complete regression of vegetations in less than 24 days and the absence of recurrence at one-year follow-up.

Sheng Wu Yi Xue Gong Cheng Xue Za Zhi, 1999 Dec, 16(4), 411 - 4
{Modification of the surface of prosthetic valve material by LGDP for protection against microbial adhesion}; Wan C et al.; This study aimed at the surface modification of prosthetic valve materials by means of cold glow discharge plasma . Orthogonal design was adopted to select the optimal scheme by the experiments of the singular factor, such as output power, gas flow rate, treatment time etc . The PEG-200 was introducted to the surface . Grafting a layer of heparin onto the surface; coating a layer of PEG-like onto the surface first, and then the grafting heparin . Comparing the adhesive amounts of staphylococcus epidermidis (SE), the adhesive curves for Dacron, which was treated by the above methods; the last one is the best.

Biochemistry, 2003 Feb 4, 42(4), 1118 - 28
Changes in stability upon charge reversal and neutralization substitution in staphylococcal nuclease are dominated by favorable electrostatic effects; Schwehm JM et al.; Single site mutations that reverse or neutralize a surface charge were made at 22 ionizable residues in staphylococcal nuclease . Unfolding free energies were obtained by guanidine hydrochloride denaturation . These data, in conjunction with previously obtained stabilities of the corresponding alanine mutants, unequivocally show that the dominant contribution to stability for virtually all of the wild-type side chains examined is the electrostatic effect associated with each residue's charged group . With only a few exceptions, these charges stabilize the native state, with an average loss of 0.5 kcal/mol of stability upon neutralization of a charge . When the charge is reversed, the average destabilization is doubled . Structure-based calculations of electrostatic free energy with the continuum method based on the finite difference solution to the linearized Poisson-Boltzmann equation reproduce the observed energetics when the polarizability in the protein interior is represented with a dielectric constant of 20 . However, in some cases, large differences are found, giving insight into possible areas for improvement of the calculations . In particular, it appears that the assumptions made in the calculations about the absence of electrostatic interactions in the denatured state and the energetic consequences of dynamic fluctuations in the native state will have to be further explored.

Trans Am Ophthalmol Soc, 2002, 100, 137 - 41; discussion 141-2
Bacterial resistance after short-term exposure to antibiotics; Wood TO et al.; PURPOSE: To determine if there is a difference in antibiotic sensitivity to coagulase-negative Staphylococcus (CNS) cultured from the host versus the donor cornea at the time of corneal transplantation . Then to apply this knowledge to preoperative preparation of patients undergoing eye surgery . METHOD: A total of 923 donor corneas stored in Optisol and 895 host corneas with no preoperative antibiotic exposure were cultured . Forty-two CNS positive cultures grew from the donor corneas and 40 from the host corneas (P = .5) . RESULTS: There was an increase in resistance in the bacteria cultured from the donor compared with the host . The most striking changes occurred in host versus donor to: ciprofloxacin 27.5% (P = .0033); gentamicin 27% (P = .0113); tobramycin 31.6% (P = .059) . The combination of polymixin, bacitracin, and neomycin (P/B/N) was significantly better than ciprofloxacin, gentamicin, and tobramycin or the combination of ciprofloxacin, gentamicin, and tobramycin (C/G/T) (P = .0007) . CONCLUSION: The combination of C/G/T exhibited the highest change to resistant bacteria . P/B/N was the most effective commercially available preparation . These results should be considered when making the decision about which preoperative antibiotic to use, if any.

J Paediatr Child Health, 2003 Jan-Feb, 39(1), 73 - 4
Iatrogenic injury in childhood staphylococcal scalded skin syndrome; Holme SA et al.; Staphylococcal scalded skin syndrome (SSSS) may cause significant morbidity in children . It is common practice for adhesive occlusive dressings to be used to apply topical local anaesthetic prior to venepuncture . We report two cases in which removal of these dressings from children with SSSS caused injury and discomfort in areas previously free from blistering . We recommend that an alternative method of topical anaesthetic application is used to minimize skin trauma in these patients.

Scand J Immunol, 2003 Jan, 57(1), 62 - 7
Staphylococcal enterotoxin B upregulates fas-mediated apoptosis of peripheral blood mononuclear cells in childhood atopic dermatitis; Sohn MH et al.; Staphylococcal infection-producing superantigens, such as staphylococcal enterotoxin B (SEB), are presumed to play an important role of inflammatory processes in atopic dermatitis (AD) . The aim of this study was to elucidate the apoptotic response of peripheral blood mononuclear cells (PBMCs) from children with AD . PBMCs from AD children were sampled and cultured with SEB stimulation . Levels of apoptosis and Fas expression were measured using flow cytometry; the soluble Fas ligand (sFasL) was also measured using the enzyme-linked immunosorbent assay method, and the expression of FasL in PBMCs was observed using reverse transcriptase-polymerase chain reaction . There was no difference in the initial levels of apoptosis and Fas expression in precultured PBMCs of AD patients and healthy donors . After culturing for 48 h under SEB stimulation, the apoptosis level and Fas expression were significantly upregulated in the PBMCs from AD children compared with that from the normal controls . In patients, the sFasL was significantly increased, and the expression of FasL was observed in messenger RNA of peripheral monocytes . These results suggest that the Fas/FasL system is involved in the apoptosis induced by SEB in AD, with simultaneous increases in sFasL and expression of FasL.

Am J Perinatol, 2002 Nov, 19(8), 435 - 8
Parotid swelling in a premature neonate; Chevalier J et al.; Suppurative parotitis in neonates is very rare . We report a case of unilateral suppurative parotitis in a 29-week gestation age infant, who recovered completely following medical treatment . Diagnosis was by clinical examination and microbiology, and parotid involvement confirmed by computed tomographic (CT) scan . The causative organism was coagulase negative Staphylococcus epidermidis.

J Immunol, 2003 Feb 1, 170(3), 1406 - 15
Human intestinal epithelial cells are broadly unresponsive to Toll-like receptor 2-dependent bacterial ligands: implications for host-microbial interactions in the gut; Melmed G et al.; Intestinal epithelial cells (IEC) interact with a high density of Gram-positive bacteria and are active participants in mucosal immune responses . Recognition of Gram-positive organisms by Toll-like receptor (TLR)2 induces proinflammatory gene expression by diverse cells . We hypothesized that IEC are unresponsive to Gram-positive pathogen-associated molecular patterns and sought to characterize the functional responses of IEC to TLR2-specific ligands . Human colonic epithelial cells isolated by laser capture microscopy and IEC lines (Caco-2, T84, HT-29) were analyzed for expression of TLR2, TLR6, TLR1, and Toll inhibitory protein (Tollip) mRNA by RT-PCR and quantitative real-time PCR . Response to Gram-positive bacterial ligands was measured by NF-kappa B reporter gene activation and IL-8 secretion . TLR2 protein expression was analyzed by immunofluorescence and flow cytometry . Colonic epithelial cells and lamina propria cells from both uninflamed and inflamed tissue demonstrate low expression of TLR2 mRNA compared with THP-1 monocytes . IECs were unresponsive to TLR2 ligands including the staphylococcal-derived Ags phenol soluble modulin, peptidoglycan, and lipotechoic acid and the mycobacterial-derived Ag soluble tuberculosis factor . Transgenic expression of TLR2 and TLR6 restored responsiveness to phenol soluble modulin and peptidoglycan in IEC . In addition to low levels of TLR2 protein expression, IEC also express high levels of the inhibitory molecule Tollip . We conclude that IEC are broadly unresponsive to TLR2 ligands secondary to deficient expression of TLR2 and TLR6 . The relative absence of TLR2 protein expression by IEC and high level of Tollip expression may be important in preventing chronic proinflammatory cytokine secretion in response to commensal Gram-positive bacteria in the gut.

Jpn J Antibiot, 2002 Oct, 55(5), 500 - 13
{A questionnaire survey on the theory of postoperative infection prophylaxis in urology}; Shinagawa N et al.; A questionnaire survey on the theory of postoperative infection prophylaxis was conducted to obtain the consensus on perioperative antimicrobial use among urologists in Japan in the period from April to July 2000 . Sixty-three of the 87 urologists replied, and the following consensus was obtained . An antimicrobial prophylaxis (AMP) agent should be chosen based on their efficacy against the pathogens expected to be contaminants, such as Escherichia coli, Staphylococcus spp., Klebsiella pneumoniae and Bacteroides fragilis group . Use an AMP agent that achieves a bactericidal concentrations in both the serum and operating site . Use an AMP agent that has little unfavourable side effects . The newer agents should be considered as a therapeutics for postoperative infections . The therapeutic antimicrobial agents having no cross-resistance to the AMP agents should be used, if postoperative infection is suspected or developed . The most commonly used agent for clean operations are cefazolin (CEZ), followed by cefotiam (CTM) and piperacillin (PIPC), in this order . For clean-contaminated operations, the most commonly used agent is CTM, followed by PIPC and CEZ.

Indian J Ophthalmol, 2002 Dec, 50(4), 295 - 9
Microbiological study of neonatal conjunctivitis with special reference to Chlamydia trachomatis; Mohile M et al.; PURPOSE: To study the microbial agents, chiefly Chlamydia trachomatis and other bacteria, in neonatal conjunctivitis . METHODS: Conjunctival specimens from 70 newborns with conjunctivitis were subjected to bacterial culture and sensitivity testing, monoclonal antibody based C . trachomatis antigen detection test and species-specific Chlamydia antibody detection in the sera of babies and their mothers, by micro-immunofluorescence assay . RESULTS: Bacteria were isolated from 35 (50%) babies; the majority (20, 57.14%) were Staphylococcus epidermidis . C . trachomatis antigen was detected in conjunctival smears of 17 (24%) babies, and 6 (35.29%) of them were positive for other bacteria . Six babies and their mothers tested positive for C . trachomatis Ig G antibodies . At follow-up after 14 weeks, 6 (35.29%) of the Chlamydia antigen-positive babies were found to have developed recurrent conjunctivitis . CONCLUSION: C . trachomatis is responsible for almost a quarter of all cases of neonatal conjunctivitis, with recurrences in 35% of cases . Bacteria could be isolated from 50% of the patients though the exact role of Staphylococcus epidermidis, isolated from 28.65% of the neonatal conjunctivitis cases, remains unclear.

Cell Mol Life Sci, 2002 Nov, 59(11), 1993 - 8
Peptide aptamers: exchange of the thioredoxin-A scaffold by alternative platform proteins and its influence on target protein binding; Klevenz B et al.; Peptide aptamers have emerged as powerful new tools for molecular medicine . They can specifically bind to and functionally inactivate a given target molecule under intracellular conditions . Typically, peptide aptamers are generated by screening a randomized peptide expression library, displayed from the Escherichia coli thioredoxin A (TrxA) protein . Here, we transferred peptide moieties from defined TrxA-based peptide aptamers to alternative scaffold proteins, such as the green fluorescent protein and staphylococcal nuclease . Yeast and mammalian two-hybrid assays as well as in vitro binding analyses show that the TrxA scaffold can be a major determinant for the binding of peptide aptamers . In addition, we demonstrate that TrxA can correctly display peptide sequences that correspond to the binding domains of natural interaction partners . Therefore, sequence analyses of TrxA-based peptide aptamers, isolated by two-hybrid screening from randomized expression libraries, should also be useful to find cellular binding partners for a given target protein, by homology.

J Virol, 2003 Feb, 77(3), 2093 - 104
Antibody-mediated targeting of an adenovirus vector modified to contain a synthetic immunoglobulin g-binding domain in the capsid; Volpers C et al.; Adenovirus vectors have been targeted to different cell types by genetic modification of the capsid or by using recombinant or chemically engineered adaptor molecules . However, both genetic capsid modifications and bridging adaptors have to be specifically tailored for each particular targeting situation . Here, we present an efficient and versatile strategy allowing the direct use of monoclonal antibodies against cell surface antigens for targeting of adenovirus vectors . A synthetic 33-amino-acid immunoglobulin G (IgG)-binding domain (Z33) derived from staphylococcal protein A was inserted into the adenovirus fiber protein . The fiber retained the ability to assemble into trimers, bound IgG with high affinity (Kd = 2.4 nM), and was incorporated into vector particles . The transduction efficiency of the Z33-modified adenovirus vector in epidermal growth factor receptor (EGFR)-expressing cells was strongly and dose-dependently enhanced by combination with an EGFR-specific monoclonal antibody . The antibody-mediated increase in cellular transduction was abolished in the presence of competing protein A . In targeting experiments with differentiated primary human muscle cells, up to a 77-fold increase in reporter gene transfer was achieved by preincubation of the vector with monoclonal antibodies directed against neuronal cell adhesion molecule or integrin alpha(7), respectively . The IgG-binding adenovirus vector holds promise for directed gene transfer to a wide variety of cell types by simply changing the target-specific antibody.

Circ J, 2003 Jan, 67(1), 3 - 7
Changing spectrum of infective endocarditis: review of 194 episodes over 20 years; Ako J et al.; A review of admission records identified 194 episodes of infective endocarditis (IE) from January 1980 to December 1999 at a community hospital in Tokyo . The cases were divided into decades, and the clinical picture and short-term outcomes were compared and analyzed . The mean age of patients in the 1990s was older (45.5 +/-13.2 vs 55.1+/-12.6 years, p<0.001), and prosthetic valve endocarditis was significantly more frequently seen (14.4% vs 31.8%, p=0.004) . None had a history of intravenous drug abuse (IVDA) . Patients on chronic hemodialysis comprised 5.8% of IE cases in the 90s . Overall, dental procedure or caries still remained the main presumed source of infection . Staphylococcal IE showed a tendency to increase, and methicillin-resistant staphylococcal IE was significantly prominent in the 90s (0% vs 10.4%, p=0.0006) . The overall in-hospital mortality was similar between the 2 groups (13.6% vs 18.8%, NS) . Multivariate analysis found neurological abnormality, renal insufficiency and staphylococcal IE as predictors of in-hospital mortality . The characteristics of IE in Japan have changed, even among non-IVDA patients, and it appears to occur in a more high-risk patient population, which may warrant a more aggressive therapeutic approach to its management and treatment.

J Clin Microbiol, 2003 Jan, 41(1), 519 - 20
Staphylococcus lugdunensis carrying the mecA gene causes catheter-associated bloodstream infection in premature neonate; Tee WS et al.; A premature neonate had a catheter-associated bloodstream infection due to Staphylococcus lugdunensis . The MIC of oxacillin for the strain was >256 microg/ml, and the mecA gene of S . lugdunensis was detected by PCR . The infection was resolved after removal of the line and treatment with vancomycin for 2 weeks.

Child Care Health Dev, 2002 Sep, 28 Suppl 1, 27 - 9
Evidence for a genetic component in sudden infant death syndrome; Gordon AE et al.; There is increasing evidence that inflammatory responses have been elicited in some Sudden Infant Death Syndrome (SIDS) infants and that these responses are under genetic control . The objective of this study was to investigate the hypothesis that the cytokine responses of SIDS parents (n = 41) differed significantly from control donors (n = 61) . Blood samples were stimulated with the staphylococcal toxin TSST-1 and LPS from Eschericia coli and assessed for production of TNF, IL-1, IL-6, IFN and IL-10 . In response toTSST-1 (P < 0.02) and LPS (P < 0.002), SIDS parents produced higher levels of IL-1 than the controls . SIDS parents produced higher levels of IFN in response to TSST-1 compared to LPS (P < 0.001) although in response to LPS, the IFN (P = 0.0008) and IL-6 (P < 0.0002) responses of the SIDS parents were lower than those of the controls . For TNF and IL-10, there was little difference between the two groups unless the effect of smoking was considered . As part of this work, a small pilot genotyping study was carried out using DNA from SIDS parents (n = 10), control donors (n = 10) and Bangladeshi subjects (n = 10) . An IFN polymorphism (3/3) was found in 40%,15.4% and 0% of donors respectively . Staphylococcal toxins have been identified in SIDS infants therefore this study highlights the importance of assessing IL-1 levels . Determination of cytokine polymorphisms and consideration of interactions between these and environmental factors such as smoking in high, average and low risk ethnic groups will assist in establishing the contribution of these factors to an infant's susceptibility to SIDS.

Altern Lab Anim, 2002 Nov-Dec, 30(6), 581 - 95
Evaluation and prevalidation of an immunotoxicity test based on human whole-blood cytokine release; Langezaal I et al.; Immunotoxicology is a relatively new field in toxicology, and is one of emerging importance, because immunotoxicity appears to contribute to the development of cancer, autoimmune disorders, allergies and other diseases . At present, there is a lack of human cell-based immunotoxicity assays for predicting the toxicity of xenobiotics toward the immune system in a simple, fast, economical and reliable way . Existing immunotoxicity tests are mainly performed in animals, although species differences favour human-based testing . Whole-blood cytokine release models have attracted increasing interest, and are broadly used for pharmacological in vitro and ex vivo studies, as well as for pyrogenicity testing . We have adapted those methods for immunotoxicity testing, to permit the potency testing of immunostimulants and immunosuppressants . Following stimulation with a lipopolysaccharide or staphylococcal enterotoxin B, monocytes and lymphocytes release interleukin-1beta and interleukin-4, respectively . Thirty-one pharmaceutical compounds, with known effects on the immune system, were used to optimise and standardise the method, by analysing their effects on cytokine release . The in vitro results were expressed as IC50 values for immunosuppression, and SC(4) (fourfold increase) values for immunostimulation, and compared with therapeutic serum concentrations of the compounds in patients, and in vivo LD50 values from animal studies . The in vitro results correlated well with the in vivo data, so the test appears to reflect immunomodulation . Results were reproducible (CV = 20 +/- 5%), and the method could be transferred to another laboratory (r(2) = 0.99) . We therefore propose this method for further validation and for use in immunotoxicity testing strategies.

Zhongguo Zhong Yao Za Zhi, 2000 May, 25(5), 303 - 5
{Action of fengsuidan granules on experimental anti-oral ulcer}; Wang SR et al.; OBJECTIVE: To explore the anti-oral ulcer action, anti-inflammatory and analgesic effect of Fengsuidan Granules(FSDG) . METHODS: FSDG(0.5, 5.0, 10.0 g.kg-1) were administered to experimental animals . The oral ulcer in experimental animals was effected by means of white staphylococcus (sc) and carbolic acid (buming) . RESULTS: Such as acetic acid body turning, hot-plate, auricle inflammation by dimethylbenzene and foot swelling by egg white were used . CONCLUSION: Following certain regularities of dose-effect relationship, FSDG has anti-oral ulcer effect, helps reduce and heal the ulcer and works efficaciously in analgesic and anti-inflammatory cases.

Zh Mikrobiol Epidemiol Immunobiol, 2002 Nov-Dec, (6), 52 - 5
{Effect of purified staphylococcal toxoid preparation on the in vitro production of interferon by leukocytes from patients with atopic dermatitis}; Mokronosova MA et al.; The impact of purified staphylococcal toxoid (PST) on the in vitro production of interferon (IFN) by blood leukocytes was evaluated . PST was found to produce a stimulating effect on the production of alpha-IFN in patients with atopic dermatitis (AD): in 88% of cases a two-fivefold increase in IFN production was observed in AD patients in comparison with healthy donors . In addition, the incubation of leukocytes obtained from atopic and nonatopic patients with PST was shown to stimulate the production of gamma-IFN by these leukocytes fivefold, on the average . These results give theoretical basis for use of PST (as an inducer of endogenic IFN) for AD patients treatment.

Toxicology, 2003 Mar 14, 185(1-2), 161 - 74
Effects of a superantigen-antibody recombinant fusion protein (r-C242 Fab-SEA) on toxicological responses in the anaesthetised rabbit; Ilback NG et al.; The objective was to study toxin-induced effects on physiological parameters in the rabbit and whether these parameters show dose-response and co-variation after administration of a recombinant fusion protein between staphylococcal enterotoxin (SE) and the Fab fragment of an antibody . Rabbits are very sensitive to SE toxins and the cardiovascular and immune effects are similar to those observed in septic shock in man . The test compound, r-C242 Fab-SEA, was administered intravenously to anaesthetised New Zealand white rabbits at doses in the range of 0.00005-50 microg/kg . All rabbits were checked for titres of anti-SEA antibodies before entering the experiment, since they could neutralise the effect of the test compound . Heart rate, blood pressure and body temperature were continuously monitored before and during 6 h after dosing . Immediately before the start of administration and 3 and 6 h during the experiment, blood gases (pO(2) and pCO(2)), pH, haematology, clinical chemistry, cytokine response (TNF-alpha) and trace elements (Mn, Cu, Zn, Se, Ag, Cd, Hg and Pb) were measured . No mortality occurred, but at 50 microg/kg severe adverse clinical signs developed . The decrease in blood pressure was weakly dose-related . Heart rate, ECG, body temperature, pCO(2) and pH were not affected by the treatment . pO(2) tended to increase as a function of time, but not in relation to dose . WBC and PLT decreased dose dependently . TNF-alpha was not affected by the treatment . The major effects on clinical chemistry were a dose-dependent increase in AST and creatinine . Potassium and urea showed dose dependent increases, mainly at higher doses, though these changes were of less value for drug selection purposes . Trace element changes were observed, including an increase in Mn and a decrease of Zn at all doses . The Cu/Zn ratio decreased below normal at low doses, whereas at high doses in which adverse effects developed, it increased above normal . Post mortem examination revealed minimal to moderate dose-related granulocytic infiltrate in the lungs . The present study showed dose-response and co-variation between several changes in cardiovascular, haematology, clinical chemistry and trace element parameters during the initial phase of toxin-induced effects preceding a possible lethal endpoint and associated patho-physiological changes.

Folia Microbiol (Praha), 2002, 47(5), 565 - 71
Evaluation of selected features of Staphylococcus cohnii enabling colonization of humans; Waldon E et al.; Based on iron utilization, sensitivity to skin fatty acids, lipolytic and proteolytic activity the potential abilities of Staphylococcus cohnii strains to colonize humans were evaluated . The investigation included 60 strains that belong to both subspecies, viz . S . cohnii ssp . cohnii and S . cohnii ssp . urealyticus . Strains were isolated from different sources of the Intensive Care Unit and from non-hospital environment . Most of the strains were multiple antibiotic-resistant . Strains of both subspecies revealed a relatively low iron requirement . These strains were capable of utilizing iron bound in oxo acids and from host iron-binding proteins . S . cohnii ssp . urealyticus were more effective in iron uptake than S . cohnii ssp . cohnii . All investigated strains revealed sensitivity to skin fatty acids, but S . cohnii ssp . urealyticus strains were more resistant . Special features of strains of this subspecies promote colonization of humans.

Orv Hetil, 2002 Nov 10, 143(45), 2543 - 8
{Evaluation of survival in mixed connective tissue disease (MCTD)}; Bodolay E et al.; INTRODUCTION: 179 patients with mixed connective tissue disease (MCTD) were follow-up, and the cause of death was analyzed in 12 died patients . PATIENTS AND METHODS: The survival of 179 patients with MCTD was evaluated by using Kaplan-Meier's method . Clinically and immunological data of the patients were analyzed between 1 and 25 years follow-up period (mean: 13.1 +/- 5.5 years) . RESULTS: The five-year survival rate was 96.4%, 10-year survival rate was 93.9%, and the 15-year survival rate was 89.6% . The cause of death was pulmonary hypertension in 5 patients, thrombotic thrombocytopenic purpura/hemolytic uremic syndrome in 3 cases, infection in 3 cases (hepatitis C virus induced hepatic coma in 2 patients, Staphylococcus sepsis in one patient), and on one patient myocarditis . The pulmonary hypertension was the most serious prognostic factor . CONCLUSION: In patients with MCTD the pulmonary hypertension with endothelial cells proliferation and microangiopathy developed very quickly, and there was progressive and therapy resistant statement . The secondary virus and bacterial infections may develop in the patients who were followed-up the long term period . Their survival rate was better than the data in the literature . This fact may cause genetic-demographic factors, and the sequential follow-up of the patients.

J Cataract Refract Surg, 2002 Dec, 28(12), 2173 - 6
Microbial contamination of the anterior chamber during phacoemulsification; Srinivasan R et al.; PURPOSE: To study the nature and frequency of anterior chamber contamination during phacoemulsification . SETTING: Department of Ophthalmology, Jawaharlal Institute of Postgraduate Medical Education and Research, Pondicherry, India . METHODS: Eighty eyes of 80 patients having routine phacoemulsification cataract surgery were recruited into this prospective study . Bacterial cultures from the intraoperative anterior chamber aspirates from the patients were assessed . RESULTS: Anterior chamber fluid aspirates were positive for bacteria in 37 eyes (46.25%) . Coagulase-negative Staphylococcus was the most common aerobe and Propionibacterium acnes, the most common anaerobe . CONCLUSION: Results indicate that phacoemulsification has no proven advantage over conventional extracapsular cataract extraction in reducing intraoperative bacterial contamination.

J Immunol, 2003 Jan 1, 170(1), 300 - 7
Activated STAT4 and a functional role for IL-12 in human Peyer's patches; Monteleone G et al.; T cells in the Peyer's patches (PP) of the human ileum are exposed to a myriad of dietary and bacterial Ags from the gut lumen . Recall proliferative responses to common dietary Ags are readily demonstrable by PP T cells from healthy individuals, and the cytokine response is dominated by IFN-gamma . Consistent with Th1 skewing, PP cells spontaneously secrete IL-12p70, and IL-12p40 protein can be visualized underneath the PP dome epithelium . In this study, we have analyzed IL-12 signaling in PP and investigated whether IL-12 plays a functional role . CD3+ T lymphocytes isolated from PP and adjacent ileal mucosa spontaneously secrete IFN-gamma with negligible IL-4 or IL-5 . RNA transcripts for IL-12Rbeta2, the signaling component of the IL-12R, are present in purified CD4+ and CD8+ T PP lymphocytes . Active STAT4, a transcription factor essential for IL-12-mediated Th1 differentiation, is readily detectable in biopsies from PP and ileal mucosa and STAT4-DNA binding activity is demonstrable by EMSA . Nuclear proteins from CD3+ T PP lymphocytes contain STAT4 and T-bet, a transcription factor selectively expressed in Th1 cells . Stimulation of freshly isolated PP cells with staphylococcal enterotoxin B dramatically enhanced the production of IFN-gamma, an effect which was largely inhibited by neutralizing anti-IL-12 Ab . These data show that IL-12 in human PP is likely to be responsible for the Th1-dominated cytokine response of the human mucosal immune system.

Infect Immun, 2003 Jan, 71(1), 567 - 70
Induction of emetic response to staphylococcal enterotoxins in the house musk shrew (Suncus murinus); Hu DL et al.; The emetic responses induced by staphylococcal enterotoxin A (SEA), SEB, SEC2, SED, SEE, SEG, SEH, and SEI in the house musk shrew (Suncus murinus) were investigated . SEA, SEE, and SEI showed higher emetic activity in the house musk shrew than the other SEs . SEB, SEC2, SED, SEG, and SEH also induced emetic responses in this animal model but relatively high doses were required . The house musk shrew appears to be a valuable model for studying the mechanisms of emetic reactions caused by SEs.

Biophys J, 2002 Dec, 83(6), 2981 - 6
Residual charge interactions in unfolded staphylococcal nuclease can be explained by the Gaussian-chain model; Zhou HX; The discrepancy of the pH dependence of the unfolding free energy for staphylococcal nuclease from what is expected from an idealized model for the unfolded state is accounted for by the recently developed Gaussian-chain model . Residual electrostatic effects in the unfolded state are attributed to nonspecific interactions dominated by charges close along the sequence . The dominance of nonspecific local interactions appears to be supported by some experimental evidence.

J Nephrol, 2002 Nov-Dec, 15(6), 713 - 5
IgA nephropathy in a patient with chronic granulomatous disease; Narsipur SS et al.; We report the first documented case of X-linked chronic granulomatous disease (CGD) in association with IgA nephropathy in a 22 year old male with multiple soft tissue abscesses . The case highlights the potential role of staphylococcal infections in the pathogenesis of IgA nephropathy and suggests the hypothesis that the neutrophil defect in CGD predisposes to such infections which may trigger IgA immune complex formation.

Ophthalmic Res, 2002 Nov-Dec, 34(6), 349 - 56
The efficacy of intravitreal levofloxacin and intravitreal dexamethasone in experimental Staphylococcus epidermidis endophthalmitis; Yildirim O et al.; This study was designed to investigate the efficacy of intravitreal levofloxacin, and intravitreal levofloxacin and dexamethasone combined in Staphylococcus epidermidis endophthalmitis . Albino rabbits (n = 25), infected with an intravitreal inoculum of S . epidermidis (1.0 x 10(5) colony forming units/0.1 ml), were divided into five groups (n = 5) . Groups 1 and 2 received treatment 24 h after the inoculation, and groups 3 and 4 48 h after the inoculation . No treatment was given to the control group . Treatment efficacy was assessed by vitreous culture, clinical examination and histopathology . Five days after treatment, groups 1 and 2 had significantly lower clinical scores than the control group (p = 0.004, p = 0.007) . The culture results of the treatment groups were sterile . The histopathological scores of the treatment groups were lower than the control group (p = 0.007) . Studies on retinal toxicity and dose-response relation are needed to prove the efficacy of levofloxacin in S . epidermidis endophthalmitis .

Bioorg Med Chem Lett, 2003 Jan 20, 13(2), 241 - 5
Dihydropyrimidinones--a new class of anti-staphylococcal antibiotics; Brands M et al.; We report the synthesis and pharmacological evaluation of new derivatives of natural dipeptide antibiotic TAN-1057 A, B . In the course of this program, we identified novel analogues of the natural product that display similar antibacterial activity and showed improved tolerability.

Gig Sanit, 2002 Sep-Oct, (5), 42 - 4
{Residing Staphylococcal virulence as a criterion for the ecological population load}; Kartashova OL et al.; In 1996-1999, a total of 8400 children living in 6 towns and 22 districts of the Orenburg Region were surveyed for resident Staphylococcus carriage . There was a relationship between the ambient air pollution, total morbidity due to respiratory diseases, and criterion Staphylococcus carriage . It has been found that resident Staphylococcus carriage determined by the persistent characteristics of strains may be used as universal test of microbiological monitoring in the ecological status of the atmosphere.

Cell Immunol, 2002 Sep, 219(1), 28 - 37
Strong induction of tyrosine phosphorylation, intracellular calcium, nuclear transcription factors and interferongamma, but weak induction of IL-2 in naïve T cells stimulated by bacterial superantigen; Luxembourg A et al.; The outcome of T cell receptor (TCR) engagement is controlled by the differential recruitment of a variety of pathways, depending on the nature of the TCR ligand . Studies on superantigens (SAGs) were among the first describing such differential signaling; however, reported results are inconsistent . We took a quantitative approach to reinvestigate this question . Using nai;ve T cells from TCR transgenic mice, we found that compared to the antigenic peptide from pigeon cytochrome c, the SAG staphylococcal enterotoxin A very efficiently (100-2000-fold more sensitive on a weight basis) induced tyrosine kinase activity, intracellular calcium increase, and interferon (IFN)gamma production . Up-regulation of CD25 and CD69 and proliferation were less efficiently induced (20-30-fold more sensitive), and interleukin (IL)-2 production was induced least efficiently (only 2-fold more sensitive) . This differential activation profile that varies with the activation event analyzed is discussed with respect to the propensity for SAG to induce anergy.

Cytotherapy, 2002, 4(5), 395 - 406
Optimized clinical-scale culture conditions for ex vivo selective depletion of host-reactive donor lymphocytes: a strategy for GvHD prophylaxis in allogeneic PBSC transplantation; Solomon SR et al.; BACKGROUND: Ex vivo selective depletion (SD) is a strategy to prevent GvHD, in which host-reactive donor lymphocytes are selectively eliminated from a PBSC allograft while conserving useful donor immune function . Prior to testing this strategy in patients, our goal was to develop a clinical-scale SD process, which involves co-culture of donor lymphocytes and irradiated recipient cells, followed by the addition of an immunotoxin (IT) directed against the alpha-chain of the IL-2 receptor (CD25), expressed on activated donor T cells . METHODS: Stimulator cells were generated from immunomagnetically selected and expanded recipient T lymphocytes . Donor PBMCs from G-CSF-mobilized peripheral blood were co-cultured for 72 h with irradiated stimulator cells . Alloreactive T cells were targeted for elimination by the addition of the anti-CD25 IT, RFT5-SMPT-dgA, and the IT enhancer, NH(4)Cl . RESULTS: Stimulator-cell selection/expansion yielded > 2 x 10(10) highly enriched CD3(+) cells (98.9 +/- 2.2%) . After SD, cell recovery was 68.5 +/- 23.3% and viability was 84.6 +/- 6.4% . This permitted a potential T-cell dose >/= 1 x 10(8) CD3(+) cells kg(-1) to transplant recipients . Although SD donor lymphocytes retained little proliferative capacity against the original stimulator cells (2.6 +/- 0.6%), responses were conserved against third party cells (107.6 +/- 18.6%), the bacterial superantigen staphylococcus enterotoxin B (108.2 +/- 4.2%), and CMV Ag (72.1 +/- 3.8%) . DISCUSSION: We have demonstrated that ex vivo SD is feasible in clinical-scale culture conditions . The ability of this strategy to prevent GvHD is the subject of an ongoing clinical trial, in which the SD lymphocyte product is transplanted in conjunction with a T cell-depleted PBSC allograft.

Chem Rec, 2002, 2(6), 369 - 76
L-histidine decarboxylase as a probe in studies on histamine; Watanabe T et al.; Because the Falck-Hillarp formaldehyde fluorescence method, which was superbly applied to identify catecholaminergic and serotonergic neurons, is not applicable to histamine, the first author (T.W.) developed an antibody to L-histidine decarboxylase (HDC) for identification of the histaminergic neuron system in the brain . The anti-HDC antibody was of great use for mapping the location and distribution of this histaminergic neuron system . (S)-alpha-fluoromethylhistidine, a specific and potent irreversible inhibitor of HDC, was also very useful in studies on functions of the neuron system . The activity of HDC is increased by various agents, treatments, and physiological conditions . We found new compounds that increased HDC activity (i.e., tetradecanoylphobol acetate (TPA), other tumor promoters, and staphylococcal enterotoxin A); and using mast cell-deficient mutant (W/W(v)) mice, we obtained evidence that this increase occurred in macrophages . To further characterize the mechanism of increases in HDC activity, the second author (H.O.) cloned human HDC cDNA and a human HDC gene . In studies on the regulation mechanism of the HDC gene, which is expressed only in limited types of cells such as mast cells, enterochromaffin-like cells in the stomach, cells in the tuberomammillary nucleus of the brain, and macrophages, CpG islands in the promoter region of the HDC gene were found to be demethylated in cells expressing the gene, whereas they are methylated in other cells that do not express the HDC gene . In collaboration with many other researchers, we developed HDC knockout mice . The resulting research is producing a lot of interesting findings in our laboratory as well as in others . In summary, HDC has been and will be useful in studies on functions of histamine .

J Pediatr Hematol Oncol, 2002 Dec, 24(9), 710 - 3
A prospective double-blind randomized trial of urokinase flushes to prevent bacteremia resulting from luminal colonization of subcutaneous central venous catheters; Aquino VM et al.; PURPOSE: This study was undertaken to determine if central venous catheter (CVC)-related infection in children with cancer could be prevented by monthly flushing of the catheter with urokinase . PATIENTS AND METHODS: Between August 1994 and July 1998, 103 patients with cancer were randomized at the time of subcutaneous CVC placement to receive monthly flushing of their catheters with either 5000 IU of urokinase-heparin or heparin alone . Patients subsequently had blood cultures taken from their CVCs during an episode of fever . RESULTS: Seventy-four of the 103 patients (72%) enrolled in the study received at least 6 catheter flushes: 40 with urokinase-heparin and 34 with heparin . The median number of flushes was 9.5 in the urokinase-heparin group and 10.2 in the heparin-only group (P = 0.62) . There were 5 positive blood cultures in the urokinase-heparin group and seven in patients receiving heparin alone (P = 0.27) . Staphylococcus epidermidis was isolated from the blood of 3 patients receiving urokinase-heparin and 6 in those receiving heparin alone (P = 0.17) . CONCLUSION: Prophylactic monthly catheter flushes with 5000 IU urokinase did not significantly decrease the number of documented bacteremic events in children with cancer who have CVCs.

Structure (Camb), 2002 Dec, 10(12), 1619 - 26
Crystal structure of a SEA variant in complex with MHC class II reveals the ability of SEA to crosslink MHC molecules; Petersson K et al.; Although the biological properties of staphylococcal enterotoxin A (SEA) have been well characterized, structural insights into the interaction between SEA and major histocompatibilty complex (MHC) class II have only been obtained by modeling . Here, the crystal structure of the D227A variant of SEA in complex with human MHC class II has been determined by X-ray crystallography . SEA(D227A) exclusively binds with its N-terminal domain to the alpha chain of HLA-DR1 . The ability of one SEA molecule to crosslink two MHC molecules was modeled . It shows that this SEA molecule cannot interact with the T cell receptor (TCR) while a second SEA molecule interacts with MHC . Because of its relatively low toxicity, the D227A variant of SEA is used in tumor therapy.

Vopr Pitan, 2002, 71(4), 23 - 6
{Enterotoxin production as a diagnostic test in intestinal dysbacterioses }; Sheveleva SA et al.; The new approach to advancing diagnostic of dysbacterioses grounded on determination of the pathogenic factors of potential-pathogenic microflora is proposed . The method of reacting of an indirect hemagglutination with usage of erythrocital diagnosticums ostend availability of production of staphylococcal enterotoxines of types A and B in 40% of the strains of staphylococcuses, isolated at dysbacterioses . The intensity of production of staphylococcal enterotoxines for strains of microorganisms, isolated at dysbacterioses and carrying of S . aureus, and at food borne diseases, is various . The availability of clinical developments of an intestinal dysfunction in faeces of the patients contacts with persistence of enterotoxigenics S . aureus, and their elimination--with successful police of the disturbed microbiocenosis.

Invest Ophthalmol Vis Sci, 2002 Dec, 43(12), 3717 - 21
In vivo study of bacterial adhesion to five types of intraocular lenses; Kodjikian L et al.; PURPOSE: To determine in vivo behavior of the ability of the Staphylococcus epidermidis strain (American Type Culture Collection {ATCC} 14990) to attach to 120 intraocular lenses (IOLs) made of five different biomaterials: fluorine polymethylmethacrylate (PMMA), heparinized PMMA, silicone, hydrophobic acrylic, and hydrogel . The pig was chosen as an animal model of endophthalmitis, after a bibliographical analysis and a personal study of its aqueous humor composition . METHODS: Crystalline lenses from 90 domestic pigs were removed aseptically and replaced with previously infected IOLs . The animals were killed 24 hours, 72 hours, and 1 week after implantation of the IOLs . The extent of bacterial binding was then measured by counting . Results were compared with a two-factor analysis of variance (ANOVA 2), confirmed by the Kruskal-Wallis nonparametric test . RESULTS: The extent of bacterial binding (expressed as bound bacteria per area unit) was found to range in increasing order from hydrogel, to fluorine PMMA, to hydrophobic acrylic, to heparinized PMMA, to silicone polymer . Comparison of pairs of materials showed statistically significant differences, except between hydrogel and fluorine PMMA . CONCLUSIONS: To the authors' knowledge, no study has been published so far concerning the in vivo evolution of populations of bacteria adhering to different intraocular materials . Bacterial adhesion to the implant surface must therefore depend on the hydrophobicity or hydrophilicity of the biomaterial . Adhesion is also affected by the nature of the surrounding medium . Because of its complexity, the latter appears to be very difficult to model, thus making in vivo studies essential.

Eur J Pharmacol, 2002 Dec 5, 456(1-3), 123 - 31
SSR125329A, a high affinity sigma receptor ligand with potent anti-inflammatory properties; Bourrie B et al.; SSR125329A ({(Z)-3-(4-Adamantan-2-yl-3,5-dichloro-phenyl)-allyl}-cyclohexyl-ethyl-amine) is a new ligand exhibiting high affinity for sigma(1) and sigma(2) receptors and for the human Delta8-Delta7-sterol isomerase . Here we show that this molecule has potent immunoregulatory properties both in vitro and in vivo . SSR125329A inhibited staphylococcal enterotoxin B-induced mouse splenocyte proliferation in vitro, whereas in vivo it enhanced lipopolysaccharide-induced systemic release of interleukin-10 while simultaneously inhibiting tumor necrosis factor-alpha (TNF-alpha) synthesis . It also prevented graft-versus-host disease in B6D2F1 mice and protected Mrl/lpr mice against the development of its spontaneous rheumatoid-like syndrome . There is high interplay of pro- and anti-inflammatory cytokines in inflammatory processes, particularly in human rheumatoid arthritis . The results of this study provide substantial evidence that sigma receptor ligands may represent a new effective approach for rheumatoid arthritis treatment.

Pol J Vet Sci, 2002, 5(3), 197 - 202
The use of some immunomodulators and non-antibiotic drugs in a prophylaxis and treatment of mastitis; Malinowski E; Antibiotics are widely used in treatment and prophylaxis of mastitis, but their antibacterial and therapeutic efficacy continues to decrease . New methods of prophylaxis of mastitis with the use of vaccines and biological response modifiers are reviewed . Prophylactic activity of staphylococcal vaccines should be investigated in the future because of difficulties in successful treatment of infections caused by these organisms . Vaccination with J5 bacterin (0111:B4) reduced duration of IMI and local signs of clinical coliform-mastitis . Some immunomodulators proved to be effective in treatment of mastitis during lactating period . Promising results were obtained with the use of giseng saponin, herbal gel, herbal extracts, propolis, lysosubtilin, antibacterial proteins, and lysozyme dimer especially.

J Immunol Methods, 2002 Dec 20, 271(1-2), 17 - 24
Improved fluoroimmunoassays using the dye Alexa Fluor 647 with the RAPTOR, a fiber optic biosensor; Anderson GP et al.; The performance of the fluorescent dye Alexa Fluor 647 (AF647) was explored as an alternative to Cy5 for immunoassays on the RAPTOR, a fiber optic biosensor . The RAPTOR performs sandwich fluoroimmunoassays on the surface of small polystyrene optical waveguides for analyte detection . Fluorescence and immunoassay data were examined at various dye-to-protein (D/P) ratios for both Cy5 and Alexa Fluor 647 . Primarily, due to the self-quenching characteristics of Cy5, Alexa Fluor 647 is substantially more effective in fluoroimmunoassays, yielding over twice the signal for any given analyte concentration . Alexa Fluor 647 can be attached to antibodies at higher ratios, D/P=6, before self-quenching begins to limit the dye's effectiveness . Furthermore, while Alexa Fluor 647 becomes quenched at high dye-to-protein ratios, D/P=9, the net fluorescence yield reaches a maximum, as opposed to Cy5-labeled proteins, which become nearly nonfluorescent at high labeling ratios, D/P> or =6 . The limitations of Cy5 were elucidated with an immunoassay for ricin, while the advantages of Alexa Fluor 647 were demonstrated in both direct binding assays as well as in a sandwich immunoassay for staphylococcal enterotoxin B.

J Dermatol Sci, 2002 Dec, 30(3), 233 - 9
Flare-up reaction on murine contact hypersensitivity . III . Effect of staphylococcal enterotoxin B; Natsuaki M et al.; Staphylococcal enterotoxin B (SEB), a bacterial superantigen, is known as an immunomodulator because it activates an extremely large number of T-cells, and induces the production of large amounts of cytokines . In this study, we examined the effects of SEB on the contact hypersensitivity reaction (CHR) . BALB/c mice were first sensitized through haptens applied to the back, and CHR was then induced through challenge to the left ear using the same haptens . SEB was administered intravenously 4 weeks later, causing a flare-up, peaking at 24 h post-administration, in the left ear that had previously exhibited CHR . This flare-up reaction was hapten non-specific, and was inhibited by anti-mouse tumor necrosis factor (TNF)-alpha antibodies . The flare-up was also suppressed by the oral administration of cyclosporin A prior to the administration of SEB . These results suggest that SEB induces a flare-up of CHR via the production of TNF-alpha.

Res Vet Sci, 2002 Dec, 73(3), 273 - 7
Frequency of superantigen-producing Staphylococcus intermedius isolates from canine pyoderma and proliferation-inducing potential of superantigens in dogs; Hendricks A et al.; This preliminary study investigated the potential role of staphylococcal superantigens in the pathogenesis of canine pyoderma . The staphylococcal enterotoxins A (SEA), SEB, SEC and SED, and the toxic shock syndrome toxin-1 (TSST-1) were assayed in isolates from skins of dogs with pyoderma . Culture supernatants from 25 of 96 isolates were positive for multiple superantigens, with SEA and SEC being the most frequently detected . In in vitro stimulation of canine peripheral blood mononuclear cells and quantitative flow cytometry revealed that low concentrations of SEA and SEB were potent stimulators of blastogenesis of T cells.

Clin Infect Dis, 2002 Dec 1, 35(11), 1308 - 15 Epub 2002 Nov 13.
Nosocomial infections occurring during receipt of circulatory support with the paracorporeal ventricular assist system; Mekontso-Dessap A et al.; This retrospective study sought to report the spectrum of infections in a homogenous group of 39 patients who underwent implantation of the Thoratec paracorporeal ventricular assist device system (Thoratec Laboratories) in an emergency setting . Thirty-one of the 39 patients developed a total of 99 nosocomial infections (attack rate, 79.5%; incidence, 4.9 per 100 support-days) . The lungs were the most frequently involved site (31.3%), and coagulase-negative Staphylococcus species were the pathogens most frequently isolated (16.2%) . Infected patients required more transfusions and chest surgical revisions, as well as a longer duration of mechanical ventilation and a longer stay in the intensive care unit, compared with uninfected patients . Cox regression analysis revealed that chest surgical revision was the only independent risk factor for infection at any site (odds ratio, 2.6; 95% confidence interval, 1.2-5.7) . There was no significant effect of infection on heart transplantation rate and overall survival.

Clin Orthop, 2002 Nov, (404), 145 - 51
Impaction grafting and wire mesh for uncontained defects in revision knee arthroplasty; Lonner JH et al.; The current authors review the results of 17 revision total knee arthroplasties in 14 patients in whom large uncontained defects were treated with impaction allografting and molded wire mesh for containment . Knee Society clinical scores increased from an average of 47 points to 95 points and function scores increased from 48 points to 73 points at the most recent followup . No patients required revision surgery, although, nonprogressive tibial radiolucency has been observed in three patients . One patient required open reduction and internal fixation of a periprosthetic supracondylar femur fracture . One patient with an acute postoperative infection with Staphylococcus epidermidis was treated successfully with irrigation and debridement with retention of the implant . Impaction grafting with wire mesh containment for large uncontained defects in revision total knee arthroplasty is an effective method of treating massive uncontained bone loss in revision total knee arthroplasty.

J Orthop Trauma, 2002 Nov-Dec, 16(10), 696 - 700
Compressed-air power tools in orthopaedic surgery: exhaust air is a potential source of contamination; Sagi HC et al.; OBJECTIVES: To determine if the exhaust from surgical compressed-air power tools contains bacteria and if the exhaust leads to contamination of sterile surfaces . DESIGN: Bacteriologic study of orthopaedic power tools . SETTING: Level I trauma center operative theater . PATIENTS/PARTICIPANTS: None . INTERVENTION: Part I . Exhaust from two sterile compact air drills was sampled directly at the exhaust port . Part II . Exhaust from the drills was directed at sterile agar plates from varying distances . The agar plates represented sterile surfaces within the operative field . Part III . Control cultures . A battery-powered drill was operated over open agar plates in similar fashion as the compressed-air drills . Agar plates left open in the operative theater served as controls to rule out atmospheric contamination . Random cultures were taken from agar plates, gloves, drills, and hoses . MAIN OUTCOME MEASUREMENT: Incidence of positive cultures . RESULTS: In Part I, all filters from both compressed-air drill exhausts were culture negative ( = 0.008) . In Part II, the incidence of positive cultures for air drills number one and number two was 73% and 82%, respectively . The most commonly encountered organisms were, coagulase-negative Staphylococcus, and Micrococcus species . All control cultures from agar plates, battery-powered drill, gloves, and hoses were negative ( < 0.01) . CONCLUSIONS: Exhaust from compressed-air power tools in orthopaedic surgery may contribute to the dissemination of bacteria onto the surgical field . We do not recommend the use of compressed-air power tools that do not have a contained exhaust.

Laryngoscope, 2002 Nov, 112(11), 1951 - 7
The role of bone in chronic rhinosinusitis; Khalid AN et al.; OBJECTIVES: To evaluate and confirm the histological inflammatory changes that occur in bone and in the overlying mucosa in experimentally induced chronic rhinosinusitis and to evaluate differences in the inflammatory patterns that may occur with different organisms . STUDY DESIGN: Histological study of induced maxillary rhinosinusitis in 29 New Zealand White rabbits (15 with, 14 with ) 7 to 9 weeks after infection . METHODS: Following maxillary sinus ostial infection, unilateral chronic bacterial rhinosinusitis was induced in 29 New Zealand White Rabbits, using (n = 15) and (n = 14) . The pathogenic organism was confirmed by culture, and the rabbits were sacrificed at predetermined time intervals (7, 8, and 9 wk) from the time of infection . Following harvest, en bloc sinus sections were mounted, stained, and analyzed . Specific attention was given to identifying histological changes in paranasal sinus bones on both sides . RESULTS: All animals (29 of 29) demonstrated histological evidence of operative occlusion on the side of the original inoculum, and all were culture-positive for the inoculated organism at death . Histological evidence of chronic rhinosinusitis in the inoculated sinus was demonstrated in 86% of animals (25 of 29) . Evidence of chronic osteomyelitis in the noninfected side was seen in 15 of 29 animals (52%) overall, or 9 of 15 animals (60%) infected with pseudomonas and 6 of 14 (43%) animals infected with staphylococcus organisms . CONCLUSIONS: The study provides further evidence that bacterial rhinosinusitis can involve bone at a distance from the site of primary infection, thereby suggesting that infectious agents may spread through bony structures in the pathogenesis of chronic rhinosinusitis.

Ital J Anat Embryol, 2002 Jul-Sep, 107(3), 209 - 14
Bio-alcamid: a novelty for reconstructive and cosmetic surgery; Pacini S et al.; Research for new biocompatible and easily implantable materials continously proposes new molecules and new substances with biological, chemical and physical characteristics more and more adapted to aesthetic and reconstructive surgery . Bio-Alcamid (Polymekon, Italy) is a non reabsorbable polymeric material composed of alkylimide-amide groups . This cosmetic agent has been used for the treatment of 2000 patients by a multicentric trial led by different Hospitals and University in Italy and abroad . Very serious aesthetic defects such as pectus excavatum, Poland Syndrome, postoperative traumas, in addition to common aesthetic defects such as lips, cheek-bone and chin hypovolumetry and relaxing of nasolabial sulcus, have been treated by surgical implant of Bio-Alcamid . Aesthetic results were excellent; tissues felt soft and the implants were uniformly distributed . No migration or dislocation of the implants, no granulomas, no allergic response and no kind of intolerance were observed . Only 12/2000 patients had post-operative complications (Staphylococcus infections) and only 3/12 cases could be directly ascribed to the implanted material . For its characteristics Bio-Alcamid can be considered a novelty in the aesthetic and reconstructive surgery; it is absolutely biocompatible, non toxic, non allergenic, easily injectable and quickly removable . Bio-Alcamid can be defined a sort of "endoprosthetis", perfectly suitable for soft tissue augmentation and for the correction of different tissue deficiences, with a long-term safety and efficiency.

Antimicrob Agents Chemother, 2002 Dec, 46(12), 4013 - 5
Uptake and intracellular activity of linezolid in human phagocytes and nonphagocytic cells; Pascual A et al.; The intracellular penetration and activity of linezolid in human polymorphonuclear leukocytes and tissue-cultured cells (McCoy) were evaluated . Linezolid reached intracellular concentrations slightly greater than extracellular ones in both types of cell . The uptake was rapid and not saturable and was affected by environmental temperature and cell viability . Linezolid showed slight intracellular activity against Staphylococcus epidermidis at high extracellular concentrations.

FEMS Microbiol Lett, 2002 Nov 5, 216(2), 277 - 83
Characterisation of the katA gene encoding a catalase and evidence for at least a second catalase activity in Staphylococcus xylosus, bacteria used in food fermentation; Barriere C et al.; The catalase gene katA of Staphylococcus xylosus was cloned . It encodes a protein of 494 amino acids with a molecular mass of 56.9 kDa, closely related to monofunctional catalases . A katA mutant still showed a relatively high catalase activity demonstrating that S . xylosus possesses more than one enzyme . By Southern blot analysis using a katA probe, a second genetic locus distinct from katA was detected that probably contained the additional catalase gene . To analyse katA expression, a transcriptional fusion of the katA promoter region to a promoterless beta-galactosidase gene was integrated into the genome of S . xylosus . katA expression is induced upon entry into stationary phase, by oxygen and hydrogen peroxide . Iron and manganese depletion induced katA transcription . Comparing the resistance of S . xylosus wild-type and the katA mutant strain to hydrogen peroxide clearly showed that KatA is essential for S . xylosus to cope with hydrogen peroxide stress . Therefore, S . xylosus has at least two differentially expressed catalases.

FEMS Microbiol Lett, 2002 Nov 5, 216(2), 171 - 7
Regulation of icaR gene expression in Staphylococcus epidermidis; Conlon KM et al.; LightCycler and conventional reverse transcription-polymerase chain reaction (RT-PCR) were used to examine regulation of icaR, which encodes a repressor of the Staphylococcus epidermidis ica operon . Varying concentrations of NaCl and ethanol activated ica but only high levels of both compounds repressed icaR transcription . Activation of ica by subinhibitory concentrations of tetracycline, which was strain-dependent, was also associated with icaR repression . In an ICAR::Em mutant, NaCl but not ethanol activated ica whereas both compounds repressed icaR expression indicating that environmental regulation of the icaR gene is IcaR-independent . Apparently ethanol signals exclusively through IcaR to activate ica and regulates IcaR at the transcriptional and posttranscriptional levels . NaCl also regulates icaR expression but in addition can activate ica via an icaR-independent pathway.

Transfusion, 2002 Sep, 42(9), 1217 - 20
Durable response to combination therapy including staphylococcal protein A immunoadsorption in life-threatening refractory autoimmune hemolysis; Javeed M et al.; BACKGROUND: Few therapeutic options are available for severe, life-threatening, refractory autoimmune hemolytic anemia . CASE REPORT: A 53-year-old 110-kg man was seen with acute onset of symptomatic severe anemia with syncope, unstable angina, and jaundice . His nadir Hct was 8.3 percent with a peak total bilirubin of 44 mg per dL . The DAT was positive but the IAT was negative . Elution studies demonstrated an IgG pan-agglutinin antibody reactive at 37 degrees C . Treatment with high-dose corticosteroids and IVIG was instituted . An accessory spleen measuring 2 cm was identified and surgically removed, but the patient continued to have intense hemolysis . Cyclophosphamide at 200 mg per day was started . Apheresis with a staphylococcal protein A immunoadsorption column (Prosorba, Cypress Bioscience, Inc.) was initiated on Day 18 and was performed twice weekly for a total of six treatments . Cyclophosphamide was continued for a total of 14 days . His transfusion requirement ceased by the third immunoadsorption treatment . Forty units of RBCs were required over 23 days in an attempt to maintain a Hct greater than or equal to 15 percent . CONCLUSION: Refractory autoimmune hemolysis can be a life-threatening event . The patient did not achieve a response until after several different therapeutic modalities were instituted, including plasmapheresis with a staphylococcal protein A column (Prosorba) . A complete response continues to be durable for more than 1 year after therapy.

Lasers Surg Med, 2002, 31(5), 343 - 51
Effects of 810 nm laser irradiation on in vitro growth of bacteria: comparison of continuous wave and frequency modulated light; Nussbaum EL et al.; BACKGROUND AND OBJECTIVES: Low intensity laser therapy may modify growth of wound bacteria, which could affect wound healing . This study compares the effects on bacteria of 810 nm laser using various delivery modes (continuous wave or frequency modulated light at 26, 292, 1000, or 3800 Hz) . STUDY DESIGN/MATERIALS AND METHODS: Staphylococcus (S.) aureus, Escherichia (E.) coli, and Pseudomonas (P.) aeruginosa were plated on agar and then irradiated (0.015 W/cm(2); 1-50 J/cm(2)) or used as controls (sham irradiated); growth was examined after 20 hours of incubation post exposure . RESULTS: There were interactions of species and modulation frequency in the overall effects of irradiation (P = 0.0001), and in the radiant exposure mediated effects (P = 0.0001); thus individual frequencies and each bacterium were analysed separately . Bacteria increased following 3800 Hz (P = 0.0001) and 1000 Hz (P = 0.0001) pulsed irradiation; at particular radiant exposures P . aeruginosa proliferated significantly more than other bacteria . Pulsed laser at 292 and 26 Hz also produced species-dependent effects (P = 0.0001; P = 0.0005); however, the effects for different radiant exposures were not significant . Bacterial growth increased overall, independent of species, using continuous mode laser, significantly so at 1 J/cm(2) (P = 0.02) . Analysis of individual species demonstrated that laser-mediated growth of S . aureus and E . coli was dependent on pulse frequency; for S . aureus, however, there was no effect for different radiant exposures . Further tests to examine the radiant exposure effects on E . coli showed that growth increased at a frequency of 1000 Hz (2 J/cm(2); P = 0.03) . P . aeruginosa growth increased up to 192% using pulsed irradiation at 1000-3800 Hz; whereas 26-292 Hz laser produced only a growth trend . CONCLUSIONS: The findings of this study point to the need for wound cultures prior to laser irradiation of infected wounds . Similar investigations using other common therapeutic wavelengths are recommended .

Int J Mol Med, 2002 Dec, 10(6), 695 - 9
Differential pathomechanisms of epidermal necrolytic blistering diseases; Paquet P et al.; Staphylococcal scalded skin syndrome (SSSS) results from the effect of exfoliative-toxins produced by staphylococcal strains . The disease affects predominantly children, and is rare in adults . We report two cases of the adult type of SSSS . Corticotherapy, chronic alcohol abuse and epilepsy-related immune changes might have been predisposing factors in these patients . The immunopathological characteristics of the inflammatory cell infiltrate in adults SSSS have not been thoroughly explored so far in the literature . Biopsies from 2 patients with bullous SSSS skin were studied by means of immunochemistry using a panel of 10 antibodies directed to FXIIIa, CD15, CD31, CD45R0, CD50, CD54, CD62E, CD95, CD106, and L1-protein, respectively . Cutaneous biopsies from related blistering diseases were compared . They included drug-induced toxic epidermal necrolysis (TEN), bullous impetigo and superficial pemphigus . A dense cell infiltrate composed of granulocytes (CD15+), macrophages (L1 protein+) and memory T cells (CD45R0+) and a strong expression of ICAM-3 (CD50) were present in the epidermis . CD95+ keratinocytes were lining the intraepidermal blisters . Type I dermal dendrocytes (Factor XIIIa+) were numerous and plump in the dermis . Bullous impetigo exhibited the same pattern of inflammatory cells, but with a lower density in type I dermal dendrocytes . TEN differed from SSSS by both the absence of CD15+ granulocytes and a stronger expression of the pro-apoptotic CD95 antigen in the epidermis . In superficial pemphigus, CD95 antigen was not expressed, and CD15+ granulocytes, CD45R0+ lymphocytes and L1 protein+ monocytes were much less numerous . It is concluded that the specific binding of SSSS-induced exotoxins to the desmosomes alters the keratinocyte metabolism leading to an inflammatory reaction followed by focal apoptosis . Our findings are in line with the concept that SSSS exotoxins might be superantigens . A common pathomechanism leading to epidermal destruction is likely operative in SSSS and bullous impetigo . The inflammatory cell composition in TEN and superficial pemphigus markedly differs from that in SSSS.

J Surg Res, 2002 Oct, 107(2), 196 - 202
Dendritic cell-tumor cell fusion and staphylococcal enterotoxin B treatment in a pancreatic tumor model; McConnell EJ et al.; BACKGROUND: Surgical resection of pancreatic tumors removes gross disease but not metastases . Adjuvant therapy such as chemotherapy and radiation treatment is of little value in metastatic pancreatic cancer . The hypothesis of this investigation is that specific and effective immunotherapeutic vaccine (dendritic/tumor cell fusion) will activate cytotoxic T lymphocytes (CTLs), leading to the eradication of spontaneous pancreatic cancer . METHODS: We have developed a double transgenic mouse model (MET) that forms spontaneous pancreatic tumors and expresses the human MUC1 antigen . Seven-week-old MET mice (n = 8) were treated every 3 weeks with the vaccine . In addition, these mice received 50 microg of superantigen staphylococcal enterotoxin B (SEB), a known T cell stimulant, prior to the first vaccination . A second treatment group received SEB alone (n = 8) and controls received no treatment (n = 9) . MUC1-specific CTLs were measured by chromium release assay . At 10 weeks of age and at necropsy, MUC1 serum levels were measured using a MUC1-specific ELISA . RESULTS: Mice were known to harbor microscopic foci of cancer at birth . Survival was enhanced in vaccine as well as SEB-treated mice (75% CI +/- 0.42) compared to controls (11% CI +/- 0.28) and both groups of treated mice exhibited mature CTLs without in vitro stimulation . MUC1 serum levels of the vaccine group were 50% less than that of control (P < 0.04) at 10 weeks . MUC1 serum levels directly correlated with tumor weight at necropsy (r = 0.86) . CONCLUSIONS: This is the first evidence that MUC1-specific CTLs can be stimulated to enhance survival in a spontaneous tumor model.

Biochemistry, 2002 Nov 19, 41(46), 13791 - 7
Robustness of the long-range structure in denatured staphylococcal nuclease to changes in amino acid sequence; Ackerman MS et al.; A nativelike low-resolution structure has been shown to persist in the Delta 131 Delta denatured fragment of staphylococcal nuclease, even in the presence of 8 M urea . In this report, the physical-chemical basis of this structure is addressed by monitoring changes in structure reflected in residual dipolar couplings and diffusion coefficients as a function of changes in amino acid sequence . Ten large hydrophobic residues, previously shown to play dominant roles in the stability of the native state, are replaced with polar residues of similar shape . Modest increases in the Stokes radius determined by NMR methods result from replacement of five isoleucine/valine residues with threonine, one leucine with glutamine, and oxidation of four methionines to the sulfoxides . Yet in the presence of all ten hydrophobic to polar substitutions and 8 M urea, the NMR signature of a native-like topology is still largely intact . In addition, removal of 30 residues from either the N-terminus (which deletes a three-strand beta meander) or C-terminus (a long extended segment and the final alpha helix) produces only very small changes in long-range structure . These data indicate that both the general shape of the denatured state and the angular relationships of individual bond angles to the axes describing the spatial distribution of the protein chain are insensitive to large changes in the amino acid sequence, a finding consistent with the conclusion that the long-range structure of denatured proteins is encoded primarily by local steric interactions between side chains and the polypeptide backbone.

J Bacteriol, 2002 Dec, 184(23), 6624 - 34
The nitrate reductase and nitrite reductase operons and the narT gene of Staphylococcus carnosus are positively controlled by the novel two-component system NreBC; Fedtke I et al.; In Staphylococcus carnosus, the nreABC (for nitrogen regulation) genes were identified and shown to link the nitrate reductase operon (narGHJI) and the putative nitrate transporter gene narT . An nreABC deletion mutant, m1, was dramatically affected in nitrate and nitrite reduction and growth . Transcription of narT, narGHJI, and the nitrite reductase (nir) operon was severely reduced even when cells were cultivated anaerobically without nitrate or nitrite . nreABC transcripts were detected when cells were grown aerobically or anaerobically with or without nitrate or nitrite . NreA is a GAF domain-containing protein of unknown function . In vivo and in vitro studies showed that NreC is phosphorylated by NreB and that phospho-NreC specifically binds to a GC-rich palindromic sequence to enhance transcription initiation . This binding motif was found at the narGHJI, nir, and narT promoters but not at the moeB promoter . NreB is a cytosolic protein with four N-terminal cysteine residues . The second cysteine residue was shown to be important for NreB function . In vitro autophosphorylation of NreB was not affected by nitrate, nitrite, or molybdate . The nir promoter activity was iron dependent . The data provide evidence for a global regulatory system important for aerobic and anaerobic metabolism, with NreB and NreC forming a classical two-component system and NreB acting as a sensor protein with oxygen as the effector molecule.

Liver Transpl, 2002 Nov, 8(11), 1014 - 9
Detection of hepatitis C virus sequences in brain tissue obtained in recurrent hepatitis C after liver transplantation; Vargas HE et al.; Patients with chronic hepatitis C frequently report tiredness, easy fatigability, and depression . The aim of this study is to determine whether hepatitis C virus (HCV) replication could be found in brain tissue in patients with hepatitis C and depression . We report two patients with recurrent hepatitis C after liver transplantation who also developed severe depression . One patient died of multiorgan failure and the other, septicemia caused by Staphylococcus aureussis . Both patients had evidence of severe hepatitis C recurrence with features of cholestatic fibrosing hepatitis . We were able to study samples of their central nervous system obtained at autopsy for evidence of HCV replication . The presence of HCV RNA-negative strand, which is the viral replicative form, was determined by strand-specific Tth-based reverse-transcriptase polymerase chain reaction . Viral sequences were compared by means of single-strand conformation polymorphism and direct sequencing . HCV RNA-negative strands were found in subcortical white matter from one patient and cerebral cortex from the other patient . HCV RNA-negative strands amplified from brain tissue differed by several nucleotide substitutions from serum consensus sequences in the 5' untranslated region . These findings support the concept of HCV neuroinvasion, and we speculate that it may provide a biological substrate to neuropsychiatric disorders observed in patients with chronic hepatitis C . The exact lineage of cells permissive for HCV replication and the possible interaction between viral replication and cerebral function that may lead to depression remain to be elucidated.

Folia Microbiol (Praha), 2002, 47(4), 441 - 4
Chlamydial infection of cats and human health; Travnicek M et al.; Chlamydia psittaci var . felis is considered as a primary and important agent in the etiology of infectious diseases of the upper respiratory tract and eyes in cats, having zoonotic potential . We investigated 13 cats aged between 2 months and 7 years, in which conjunctivitis, rhinitis, laryngotracheitis, bronchopneumonia and lymph adenopathy was clinically diagnosed . To detect the antigen of C . psittaci the Clearview Chlamydia Direct Test was used for the first time . The presence of C psittaci in 10 of 13 investigated cats and in 8 nasal mucosa smears from 10 investigated cats were confirmed in conjunctiva . Using bacteriological examination different species of the genus Staphylococcus in conjunctiva from all 13 investigated animals were also confirmed . Serological investigation using complement fixation test was negative in all animals.

J Vasc Surg, 2002 Nov, 36(5), 1027 - 30
Temporin A as a prophylactic agent against methicillin sodium-susceptible and methicillin sodium-resistant Staphylococcus epidermidis vascular graft infection; Ghiselli R et al.; OBJECTIVE: The purpose of this study was to investigate the efficacy of temporin A as a prophylactic agent in a rat model of vascular graft infection from methicillin sodium-susceptible and methicillin sodium-resistant Staphylococcus epidermidis . METHODS: The prospective, randomized, controlled animal study set in a research laboratory in a university hospital used 280 adult male Wistar rats (weight range, 280 to 350 g) . Graft infections were established in the back subcutaneous tissue of rats with implantation of 1-cm(2) sterile Dacron grafts followed by topical inoculation with 2 x 10(7) colony-forming units of S epidermidis . The study for each staphylococcal strain included: one control group (no graft contamination), one contaminated group that did not receive any antibiotic prophylaxis, one contaminated group that received temporin A-soaked graft, two contaminated groups that received perioperative intraperitoneal cefazolin (30 mg/kg) or vancomycin hydrochloride prophylaxis (10 mg/kg), and two contaminated groups that received temporin A-soaked graft and perioperative intraperitoneal cefazolin (30 mg/kg) or vancomycin hydrochloride (10 mg/kg) prophylaxis . All grafts were explanted at 7 days after implantation . The main outcome measure was quantification of bacterial contamination . RESULTS: Overall, the perioperative prophylaxis based on soaked grafts was not significantly different to that of parenteral vancomycin hydrochloride . Only the combination between temporin A and vancomycin hydrochloride produced a complete bacterial inhibition for both strains . CONCLUSION: Temporin A showed a similar antibacterial in vitro activity against the two different strains . The in vivo results suggest its potential use in providing prophylaxis to direct graft contamination when used in combination with parenteral vancomycin hydrochloride.

Transfusion, 2002 Nov, 42(11), 1458 - 65
Hypotensive reaction during staphylococcal protein A column therapy in a patient with anomalous degradation of bradykinin and Des-Arg9-bradykinin after contact activation; Molinaro G et al.; BACKGROUND: Hypotensive reactions have occurred in patients taking angiotensin converting enzyme (ACE) inhibitors after infusion of blood previously in contact with negatively charged surfaces capable of generating kinins, which accumulate when ACE, a kininase, is inhibited . A patient with anomalous bradykinin (BK) metabolism who experienced hypotension during extracorporeal staphylococcal protein A (SPA) therapy while on an ACE inhibitor was studied . CASE REPORT: A patient with mitomycin-associated hemolytic-uremic syndrome received SPA treatments after her ACE inhibitor, lisinopril, was held . Lisinopril was restarted before her 18th SPA treatment, and immediately after return of treated plasma she developed facial redness and hypotension, which resolved after the return stopped and recurred when restarted . To study formation and degradation of kinins, exposed her plasma to glass beads . We found a normal kinin formation rate but an abnormal degradation and accumulation of Des-Arg9-BK . The kinin degradation enzymes ACE, aminopeptidase P (APP), and carboxypeptidase N (CPN) were measured while on an ACE inhibitor, showing absence of ACE activity, low APP, but normal CPN . CONCLUSION: This patient's vasodilation and hypotension during SPA therapy was associated with a pre- existing anomaly of BK metabolism . Her ACE inhibitor shifted degradation toward Des-Arg9-BK formation, and her low APP was associated with a prolonged t50 and accumulation of the vasoactive Des-Arg9-BK.

Transfusion, 2002 Nov, 42(11), 1422 - 7
Polymerized human Hb use in acute chest syndrome: a case report; Lanzkron S et al.; BACKGROUND: Acute chest syndrome (ACS) is a complication of sickle cell disease that can cause significant morbidity . Transfusion therapy has been shown to significantly increase oxygenation in patients with ACS and RBC exchange is considered the standard of care in patients at high risk of respiratory failure . CASE REPORT: A patient with ACS and several high-risk features, including thrombocytopenia, profound anemia, bilateral pulmonary infiltrates, staphylococcal sepsis, and pulmonary embolism is presented . The patient refused transfusion on religious grounds and received 12 units of human polymerized Hb solution (poly SFH-P injection, PolyHeme, Northfield Laboratories) over the course of 13 days . The patient's respiratory status improved and she was discharged home without receiving RBC transfusions . CONCLUSION: This is the first reported case that describes the use of PolyHeme in a patient with sickle cell disease, ACS, and sepsis . This therapy is thought to have been lifesaving for this patient.

Syst Appl Microbiol, 2002 Oct, 25(3), 353 - 9
Staphylococcus succinus subsp . casei subsp . nov., a dominant isolate from a surface ripened cheese; Place RB et al.; A new subspecies of the species Staphylococcus succinus, isolated from a Swiss surface ripened cheese, is described . This subspecies is differentiated from the species Staphylococcus succinus ATCC 700337T on the basis of DNA-DNA hybridisation, cell wall composition and phenotypic characteristics . Staphylococcus succinus subsp . casei could be distinguished among other things by its ability to reduce nitrate, form acid from D-mannose and D-melezitose, ferment adenosine, inosine, D-sorbitol, and 2,3-butanediol, but not D-alanine . The type strain of Staphylococcus succinus subsp . casei is DSM 15096 (CIP no . pending) . The GenBank accession numbers for the reference sequences of the 16S rDNA and the hsp60 gene used in this study are AJ320272 and AF527482, respectively.

J Am Vet Med Assoc, 2002 Apr 1, 220(7), 1025 - 7, 1006-7
Intra-abdominal botryomycosis in a dog; Share B et al.; A 6-year-old 41.8-kg (92-lb) sexually intact male German Shepherd Dog used as an attack and pursuit dog by the local police department was examined because of general malaise and sudden onset of ataxia . Abnormal findings included a high WBC count, fever, and ongoing weight loss . Physical examination was unrevealing initially, in part because of the aggressive nature of the dog . Following treatment with antibiotics, an abdominal mass was detected during a second physical examination, and was confirmed radiographically . Subsequent ultrasound examination revealed a walled mass of mixed echogenicity, with areas of detectable fluid movement . A laparotomy was performed to remove the mass, which weighed 2.2 kg (4.5 lb) . Histologic findings were consistent with botryomycosis, and Staphylococcus intermedius was isolated on bacteriologic culture . Postoperative treatment consisted of intravenous, intra-abdominal, and oral administration of antibiotics . Diagnosis of botryomycosis requires histologic examination and bacteriologic culture of the lesion . Treatment consists of surgical intervention and administration of antimicrobials . Botryomycosis is a poorly understood and rarely reported disease found in many species . A computer search of the literature failed to reveal any previous reports of intra-abdominal botryomycosis in dogs.

Biomaterials, 2003 Jan, 24(1), 27 - 34
Development of a bioluminescent ATP assay to quantify mammalian and bacterial cell number from a mixed population; Dexter SJ et al.; Modification of biomaterial surfaces is one approach aimed at improving cellular interactions and the subsequent integration of medical devices into the body . However, by optimising surfaces to enhance mammalian cell adhesion there is the potential risk that adhesion of bacteria will also be increased . Bacterial colonisation of biomaterials can be problematic as infection often results in morbidity and the consequent removal of the failing device from the body . Currently, quantifying cellular adhesion from a mixed population of bacterial and mammalian cells can only be determined by slow and laborious methods such as microscopy . ATP is a key molecule in the metabolism of both mammalian and bacterial cells and can be used to quantify cell numbers . In this study, we have modified a bioluminescence-based ATP assay to enable the differential measurement of both mammalian and bacterial cell ATP levels within the same culture . Mixed populations of Staphylococcus epidermidis and 3T3 fibroblasts were assessed both in suspension and adhered to a surface . ATP levels from cultures in suspension were selectively extracted and measured from both cell types, revealing a linear trend that would enable the differentiation of cell numbers from a mixed population . The application of the assay to adhered mixed cultures also allowed differences in ATP levels from both cell types to be distinguished . The data presented reveals that this assay would be useful for the rapid screening of cellular adhesion to modified surfaces although, its use in detecting subtle differences in ATP levels may be limited due to natural interactions between the two cell types .

Vet Surg, 2002 Nov-Dec, 31(6), 552 - 60
Removal of infected canine cemented total hip prostheses using a femoral window technique; Dyce J et al.; OBJECTIVE: To evaluate a femoral window technique for retrieval of cemented total hip prostheses . STUDY DESIGN: Retrospective clinical study . ANIMALS: Twelve dogs with infection of a cemented modular total hip prosthesis . METHODS: Implant removal was performed by an extended craniolateral approach to the hip and proximal femur without trochanteric osteotomy . The femoral cement mantle was fragmented and removed with simple orthopedic instrumentation by a lateral femoral window that was repaired using cerclage wires . Surgical technique, intraoperative and postoperative complications, bacterial culture results, histopathologic findings, and completeness of cement removal were recorded . Follow-up radiographs were taken 5 to 9 weeks postoperatively . Long-term follow-up information was obtained by client questionnaire . Functional outcome was assessed by scoring ability to stand, sit, walk, run, play, climb stairs, and get into a car . RESULTS: Prosthesis retrieval was performed 2 to 41 months after implantation (median, 14 months) . Complete removal of femoral cement was achieved in 10 dogs . A nondisplaced femoral fissure, extending proximally from the window, was an intraoperative complication in 2 dogs . Staphylococcus spp was most commonly isolated (6 dogs) from interfacial membrane samples . Systemic antibiotic therapy, dependent on susceptibility testing, was administered for 3 to 10 weeks postoperatively . There was radiographic evidence of osteotomy healing at 5- to 9-week reassessment . Recurrence of osteomyelitis was not observed . Long-term functional outcome was considered mildly abnormal . CONCLUSIONS: The lateral window was an effective technique for retrieval of retained femoral cement . CLINICAL RELEVANCE: Removal of an infected prosthesis using this technique generally resulted in a clinical outcome comparable to that with ab initio femoral head and neck excision .

J Dermatol Sci, 2002 Nov, 30(2), 129 - 35
Acne phototherapy with a high-intensity, enhanced, narrow-band, blue light source: an open study and in vitro investigation; Kawada A et al.; The purpose of this study was to investigate the efficacy of phototherapy with a newly-developed high-intensity, enhanced, narrow-band, blue light source in patients with mild to moderate acne . An open study was performed in acne patients who were treated twice a week up to 5 weeks . Acne lesions were reduced by 64% . Two patients experienced dryness . No patient discontinued treatment due to adverse effects . In vitro investigation revealed that irradiation from this light source reduced the number of Propionibacterium acnes (P . acnes), but not Staphylococcus epidermidis that were isolated from the acne patients . Phototherapy using this blue light source was effective and well tolerated in acne patients and had an ability to decrease numbers of P . acnes in vitro, suggesting that this phototherapy may be a new modality for the treatment of acne .

Eur J Pain, 2002, 6(6), 455 - 66
Treatment with staphylococcus toxoid in fibromyalgia/chronic fatigue syndrome--a randomised controlled trial; Zachrisson O et al.; We have previously conducted a small treatment study on staphylococcus toxoid in fibromyalgia (FM) and chronic fatigue syndrome (CFS) . The aim of the present study was to further assess the efficacy of the staphylococcus toxoid preparation Staphypan Berna (SB) during 6 months in FM/CFS patients . One hundred consecutively referred patients fulfilling the ACR criteria for FM and the 1994 CDC criteria for CFS were randomised to receive active drug or placebo . Treatment included weekly injections containing 0.1 ml, 0.2 ml, 0.3 ml, 0.4 ml, 0.6 ml, 0.8 ml, 0.9 ml, and 1.0 ml SB or coloured sterile water, followed by booster doses given 4-weekly until endpoint . Main outcome measures were the proportion of responders according to global ratings and the proportion of patients with a symptom reduction of > or =50% on a 15-item subscale derived from the comprehensive psychopathological rating scale (CPRS) . The treatment was well tolerated . Intention-to-treat analysis showed 32/49 (65%) responders in the SB group compared to 9/49 (18%) in the placebo group (P<0.001) . Sixteen patients (33%) in the SB group reduced their CPRS scores by at least 50% compared to five patients (10%) in the placebo group (P< 0.01) . Mean change score on the CPRS (95% confidence interval) was 10.0 (6.7-13.3) in the SB group and 3.9 (1.1-6.6) in the placebo group (P<0.01) . An increase in CPRS symptoms at withdrawal was noted in the SB group . In conclusion, treatment with staphylococcus toxoid injections over 6 months led to significant improvement in patients with FM and CFS . Maintenance treatment is required to prevent relapse.

Rinsho Ketsueki, 2002 Sep, 43(9), 828 - 32
{Acute myeloid leukemia with infective endocarditis}; Doki N et al.; We present here a patient with acute myeloid leukemia (M2) who developed fatal infective endocarditis . On admission, the patient (67-year-old male) had mitral stenosis and atrial fibrillation . Complete remission was achieved after induction chemotherapy . During the course of consolidation therapy, he developed sepsis caused by coagulase-negative staphylococcus, which was successfully treated with antibiotics . Thereafter, blood culture yielded multidrug-resistant staphylococcus epidermidis . An echocardiogram revealed mitral valve regurgitation with vegetation . He was diagnosed as having infectious endocarditis . In spite of prolonged antibiotic therapy, destruction of the mitral valve progressed, and the patient underwent valve replacement therapy . He died of cardiac tamponade 5 days after the surgery.

Br J Dermatol, 2002 Nov, 147(5), 985 - 93
Fatal bacteria granuloma after trauma: a new entity; Gao TW et al.; In the past 20 years, more than 20 cases of a type of granulomatous disease have been noticed by dermatologists in different areas of China . The patients had these features in common: (i) the lesions followed a slight trauma to the face; (ii) they were spreading dark-red plaques without pus or ulceration; (iii) new lesions appeared near to or far from the original lesion; (iv) histopathology showed histiocytic granuloma; (v) the patients had severe headache and clouding of consciousness during the later stages of the disease; (vi) all patients died within 1.5-4 years; (vii) treatment with prednisone led to some healing of the lesions but accelerated death; and (viii) all patients were from rural areas . We examined the tissues from two similar patients by electron microscopy and identified two kinds of bacteria as a possible cause of the disease . One was an anaerobic actinomycete, the other was Staphylococcus capitis . The anaerobic actinomycete was sensitive to lincomycin (a forerunner of clindamycin) . After a 5-month therapy with lincomycin, one patient survived . We infer that the cause of death is the unknown anaerobic actinomycete . Because the disease is very severe, we suggest the name 'fatal bacteria granuloma after trauma' to draw attention.

Pediatr Infect Dis J, 2002 Oct, 21(10), 984 - 6
The "surreptitious Staphylococcus": Staphylococcus lugdunensis endocarditis in a child; Sotutu V et al.; A child with congenital heart disease developed infective endocarditis caused by Staphylococcus lugdunensis . Despite an apparent excellent response to initial antibiotic treatment in clinical, inflammatory and echocardiographic indices, the patient's valve damage progressed silently and surgical intervention was required . This case highlights the potential for misidentification of S . lugdunensis, its usual susceptibility to penicillin and in particular the aggressive nature of endocarditis caused by this coagulase-negative staphylococcus . The epidemiology and treatment of endocarditis caused by this organism are reviewed.

Microb Ecol, 2002 Nov, 44(4), 365 - 71 Epub 2002 Oct 29.
Bacterial 16S rRNA gene analysis revealed that bacteria related to Arcobacter spp . constitute an abundant and common component of the oyster microbiota (Tiostrea chilensis); Romero J et al.; To explore the bacterial microbiota in Chilean oyster (Tiostrea chilensis), a molecular approach that permits detection of different bacteria, independently of their capacity to grow in culture media, was used . Bacterial diversity was assessed by analysis of both the 16S rDNA and the 16S-23S intergenic region, obtained by PCR amplifications of DNA extracted from depurated oysters . RFLP of the PCR amplified 16S rDNA showed a prevailing pattern in most of the individuals analyzed, indicating that a few bacterial species were relatively abundant and common in oysters . Cloning and sequencing of the 16S rDNA with the prevailing RFLP pattern indicated that this rRNA was most closely related to Arcobacter spp . However, analysis by the size of the amplified 16S-23S rRNA intergenic regions revealed not Arcobacter spp . but Staphylococcus spp . related bacteria as a major and common component in oyster . These different results may be caused by the absence of target for one of the primers employed for amplification of the intergenic region . Neither of the two bacteria species found in large abundance was recovered after culturing under aerobic, anaerobic, or microaerophilic conditions . This result, however, is expected because the number of bacteria recovered after cultivation was less than 0.01% of the total . All together, these observations suggest that Arcobacter-related strains are probably abundant and common in the Chilean oyster bacterial microbiota.

Appl Biochem Biotechnol, 2002 Jul-Dec, 102-103(1-6), 395 - 405
Optimization of fermentation conditions for production of glycopeptide antibiotic vancomycin by Amycolatopsis orientalis; Padma PN et al.; Glycopeptides produced by Streptomyces species are the drugs used against beta-lactam drug-resistant staphylococcal infections, and vancomycin is important among them . Increased prevalence of resistant strains increased the usage of vancomycin worldwide and also promoted attempts for indigenous production . The optimum process conditions pH, temperature, inoculum size, agitation, and aeration for vancomycin production by Amycolatopsis orientalis were evaluated, statistically analyzed, and the response surface curves were constructed . The optimum process conditions were a pH of 7.6, a temperature of 29 degrees C, an inoculum size of 4.5%, an agitation of 255 rpm, and an aeration of less than 1:10 medium-to-air ratio.

Int J Technol Assess Health Care, 2002 Summer, 18(3), 540 - 54
Effects of linezolid on hospital length of stay compared with vancomycin in treatment of methicillin-resistant Staphylococcus infections . An application of multivariate survival analysis; Willke RJ et al.; OBJECTIVES: This study was designed to estimate the effects of treatment with linezolid as compared with vancomycin, on the distribution of length of stay (LOS) for hospitalized patients with methicillin-resistant staphylococcal infections . Treatment with intravenous-oral linezolid may allow some patients to be discharged earlier than would treatment with intravenous vancomycin . METHODS: The analysis is based on the intention-to-treat sample from a randomized multinational phase 3 clinical trial of 460 patients showing that the treatments had equal efficacy . Given the nature of the LOS data, some censoring, and some imbalances between treatment groups, multivariate survival analysis was indicated . Cox proportional hazards assumptions were tested and failed, and accelerated failure time models were tested for best fit . The log-logistic model was selected and used as the basis for estimating the overall treatment effect on LOS . Two methods for multivariate corrections to the survivorship functions allowed more thorough description of the treatment effect on the distribution of LOS, including multivariate-adjusted Kaplan-Meier curves . RESULTS: The average reduction in LOS associated with linezolid treatment, based on the log-logistic model after correction for covariate effects, was 18.1% (p = .041) or 2.53 days at the median . This was consistent with differences at the medians of the adjusted survivorship functions, which were 2 or 3 days depending on the method used . Treatment-based differences exist at each decile of LOS and consistently favor linezolid . Estimated mean reduction in LOS due to linezolid was 1.62 days in both methods . CONCLUSIONS: In this study sample, linezolid treatment resulted in statistically significantly shorter hospital LOS as compared with vancomycin treatment . Appropriate use of multivariate survival analysis allows better examination of the nature of the treatment effect on LOS, which may be important for economic analysis.

Arq Bras Cardiol, 2002 Sep, 79(3), 302 - 7 Epub 2002 Oct 08.
Silent patent ductus arteriosus aneurysm; Botta AM et al.; Ductus arteriosus aneurysm, a rare and potentially fatal condition, has been reported as a complication after surgical ductus arteriosus closure . Its spontaneous appearance as a septic complication, which was common in the presurgical and preantibiotic era, has been rarely reported in the contemporary literature . Persistence of silent ductus arteriosus in healthy children and adults is a frequent condition that currently has an increasing diagnostic possibility due to the availability of more accurate investigative methods, especially echocardiography . We report the case of a 1-year-old child, in whom no previous heart disease was known, who developed a giant aneurysm of the ductus arteriosus during a staphylococcal infection . This complication appeared after craniotomy for emptying an accidental subdural hematoma . This report associates the persistence of ductus arteriosus with a complication considered rare, which has a rapidly fatal evolution.

Sheng Wu Gong Cheng Xue Bao, 2002 Jul, 18(4), 477 - 80
{The changes of rare codons increase expression level of SEA}; Shi CB et al.; The rare codons of a fragment in staphylococcal enterotoxin A gene were turned into the most high usage frequency codons in E . coli by overlap PCR technique . Genes of sea and seam were cloned into 7ZTS expression vector and transformed into JM109(DE3), respectively . The result shows that expression level of sea gene was very low, but the expression level of seam was as high as 15% of total cell proteins . The expression product shows activity of antitumor in vivo.

Antimicrob Agents Chemother, 2002 Nov, 46(11), 3621 - 3
Staphylococcus sciuri gene erm(33), encoding inducible resistance to macrolides, lincosamides, and streptogramin B antibiotics, is a product of recombination between erm(C) and erm(A); Schwarz S et al.; A gene which mediates inducible resistance to macrolides, lincosamides, and streptogramin B antibiotics, designated erm(33), was detected on the Staphylococcus sciuri plasmid pSCFS1 . Analysis of the erm(33) reading frame suggested that this gene was the product of a recombination between an erm(C) gene and an erm(A) gene . Such a recombination event is a novel observation for erm genes.

Infection, 2002 Oct, 30(5), 272 - 6
A second look at anorectal infections in cancer patients in a large cancer institute: the success of early intervention with antibiotics and surgery; Lehrnbecher T et al.; BACKGROUND: Infection of anorectal region represents a significant complication of anti-cancer therapy . Anorectal infection occurs in patients receiving aggressive chemotherapy . Untreated infection leads to substantial morbidity and in the past, mortality . METHODS: 82 episodes of anorectal infection in 64 patients with malignant diseases occurring over 12 years at the National Cancer Institute (NCI) were retrospectively reviewed . RESULTS: The overall incidence is comparable to the prior NCI experience despite a shift in patient population to a lower percentage of lymphoid/leukemic diagnoses (34% vs 77%) . There were no deaths associated with anorectal infection in the 12 years reviewed compared to seven of 44 in the previous decade (p = 0.003) . Antibiotic therapy alone was successful in managing 25/82 . Only five episodes were treated with surgery alone compared to nearly 45% in the previous decade . There were no major surgical complications . Neutropenia was present in 43/82 episodes . 11 episodes were complicated by bacteremia, predominately with Staphylococcus non- aureus (n = 8) . Wound cultures were performed in 36 episodes from 23 patients yielding 99 separate isolates . Gram-negative isolates were most common overall . Modification of antibiotic therapy with further anaerobic coverage was administered successfully in 39/77 episodes . CONCLUSION: This study illustrates that anorectal infections in cancer patients can be successfully managed with antibiotic therapy and local care; surgery can be withheld unless there is evidence of progressive infection or substantial fluctuance and necrosis.

J Food Prot, 2002 Oct, 65(10), 1628 - 31
Shelf-stable and safe intermediate-moisture meat products using hurdle technology; Kanatt SR et al.; A number of ready-to-use shelf-stable intermediate-moisture (IM) spiced mutton and spiced chicken products were developed with a combination of hurdles (reduced moisture, vacuum packing, and irradiation) . The water activity of the products was reduced to about 0.80 either by grilling or by hot-air drying . These IM products were vacuum packed and subjected to gamma radiation processing at 0 to 10 kGy . Microbiological analyses revealed a radiation dose-dependent reduction in total viable counts and in numbers of Staphylococcus species . IM meat products that did not undergo radiation treatment showed visible mold growth within 2 months . The products subjected to irradiation at 10 kGy showed an absence of viable microorganisms and also retained high sensory acceptability for up to 9 months at ambient temperatures.

J Leukoc Biol, 2002 Oct, 72(4), 657 - 67
FcgammaR cross-linking mediates NF-kappaB activation, reduced antigen presentation capacity, and decreased IL-12 production in monocytes without modulation of myeloid dendritic cell development; Drechsler Y et al.; Stimulation of monocytes (MO) through receptors for the Fc region of immunoglobulin G (FcgammaR) activates a variety of responses, including phagocytosis, antibody (Ab)-dependent cellular cytotoxicity, and production of cytokines . We previously reported that the MO subpopulation that expresses FcgammaR in high density produces high levels of tumor necrosis factor alpha (TNF-alpha) compared with FcgammaR-negative MO . Here, we show that cross-linking MO via FcgammaRI or FcgammaRII but not via FcgammaRIII activates nuclear regulatory factor-kappaB (NF-kappaB), a transcription factor involved in regulation of TNF-alpha . NF-kappaB activation peaked at 2.75 h after FcgammaRI cross-linking, involved p65 and p50 (heterodimer) and not c-rel-containing NF-kappaB complexes, and was mediated via IkappaB degradation . Cross-linking FcgammaRI, -II, as well as -III inhibited interleukin (IL)-12 (p70) production in MO, whether stimulated with Staphylococcal enterotoxin B (P<0.02) or lipopolysaccharide (P<0.02) . Inhibition of IL-12 by FcgammaR cross-linking was not mediated by TNF-alpha, as the presence of an anti-TNF-alpha Ab could not restore the reduced IL-12 production . Decreased IL-12 production correlated with reduced antigen presentation capacity (P<0.01) in the FcgammaR-cross-linked MO . Blood MO can give rise to myeloid dendritic cells (DC) . FcgammaR cross-linking did not modulate in vitro maturation of MO to fully functional myeloid DC . Allostimulatory capacity in mixed leukocyte reaction and DC marker expression (CDla, CD80, CD86) was not different between control and FcgammaRI cross-linked DC . These results suggest that signals mediated via FcgammaRI, -II, and -III have overlapping yet distinct effects on MO, which are likely to be involved in the fine-tuning of the immune responses to various stimuli.

Scand J Infect Dis, 2002, 34(9), 685 - 6
Surgical wound infection associated with Staphylococcus sciuri; Stepanovic S et al.; We describe a case of surgical wound infection due to Staphylococcus sciuri . The isolated strain was susceptible to trimethoprim-sulfamethoxazole, erythromycin, chloramphenicol, ciprofloxacin and vancomycin and resistant to gentamicin, clindamycin, rifampicin, methicillin, ampicillin and ceftriaxone . The multiresistance of the strain had a serious impact on the prolonged course of the infection . Although this bacterium is principally found in animals, our strain was probably of nosocomial origin.

Ceska Gynekol, 2002 Jul, 67(4), 210 - 5
{Occurrence and characteristics of staphylococcal enterotoxin antibodies in gynecologic patients}; Ostro A et al.; OBJECTIVE: To control the occurrence of the antibodies against Staphylococcal type A and type B enterotoxin in gynaecological patients and in selected patients to determine the thermodynamic parameters of antibodies against Staphylococcal enterotoxins in their blood samples . DESIGN: Retrospective clinical study . SETTING: 2nd Department of Obstetrics and Gynaecology, P . J . Safarik University Kosice, Slovak Republic; Research Institute of Veterinary Medicine, Kosice, Slovak Republic; Department of Microbiology and Immunology, University of Veterinary Medicine, Kosice, Slovak Republic; Department of Food Microbiology and Toxicology, Food Research Institute, University of Wisconsin, Madison, USA . METHODS: The occurrence of antibodies against Staphylococcal type A and type B enterotoxin was determined in 68 patients hospitalized in Department of Obstetrics and Gynecology in Kosice . RESULTS: The occurrence of antibodies against Staphylococcal enterotoxins was determined by radioimmunoassay (RIA) in blood samples of 45 (66%) patients . The antibodies against Staphylococcal type A enterotoxin were determined in 36 (53%) patients and the antibodies against Staphylococcal type B enterotoxin were determined in 9 (13%) patients . The antibodies against both type A and type B enterotoxins were determined simultaneously in blood samples of 10% of all patients . The thermodynamic parameters of the antibodies were determined in 5 patients with positive serum findings . CONCLUSION: With regard to the existence of heterogeneous clinical findings in large amount of patients with antibodies against Staphylococcal enterotoxins, the next study of Staphylococcal enterotoxins role in pathogenesis of wide spectrum of diseases is necessary.

Arch Toxicol, 2002 Oct, 76(10), 570 - 80 Epub 2002 Aug 06.
2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) induces Fas-dependent activation-induced cell death in superantigen-primed T cells; Camacho IA et al.; Immune response against a foreign antigen is characterized by a growth phase, in which antigen-specific T cells clonally expand, followed by a decline phase in which the activated T cells undergo apoptosis, a process termed activation-induced cell death (AICD) . In the current study, we have investigated the phase at which 2,3,7,8-tetrachlorodibenzo- p-dioxin (TCDD) acts to downregulate the antigen-specific T cell response . To this end, C57BL/6 +/+ mice were injected with staphylococcal enterotoxin A (SEA) into the footpads (10 micro g/footpad), and simultaneously treated with TCDD (10 or 50 micro g/kg intraperitoneally) . At various time points, the draining lymph node (LN) cells were analyzed for SEA-activated T cells . The data demonstrated that in C57BL/6 +/+ mice, TCDD treatment did not alter the growth phase but facilitated the decline phase of SEA-reactive T cells . TCDD caused a significant decrease in the percentage and absolute numbers of CD4(+) and CD8(+) SEA-responsive T cells expressing Vbeta3(+) and Vbeta11(+) but did not affect SEA-nonresponsive Vbeta8(+) T cells . Upon in vitro culture, TCDD-exposed SEA-immunized LN cells exhibited increased levels of apoptosis when compared with the vehicle controls . When Fas-deficient (C57BL/6 lpr/lpr) or Fas ligand defective (C57BL/6 gld/gld) mice were treated with TCDD, they failed to exhibit a decrease in percentage and cellularity of SEA-reactive T cells, thereby suggesting a role of Fas-Fas ligand interactions in the TCDD-induced downregulation of SEA-reactive T cell response . The resistance to TCDD-induced decrease in T cell responsiveness to SEA seen in Fas- and FasL-mutant mice was neither due to decreased aryl hydrocabon receptor (AhR) expression nor to altered T cell responsiveness to SEA . The current study demonstrates that TCDD does not prevent T cell activation, but prematurely induces Fas-based AICD, which may contribute to the deletion of antigen-primed T cells.

J Allergy Clin Immunol, 2002 Oct, 110(4), 617 - 23
Lactic acid bacteria inhibit TH2 cytokine production by mononuclear cells from allergic patients; Pochard P et al.; BACKGROUND: Among factors potentially involved in the increased prevalence of allergic diseases, modification of the intestinal bacteria flora or lack of bacterial stimulation during childhood has been proposed . Lactic acid bacteria (LAB) present in fermented foods or belonging to the natural intestinal microflora were shown to exert beneficial effects on human health . Recent reports have indicated their capacity to reduce allergic symptoms . OBJECTIVE: The purpose of this investigation was to determine the effect of LAB on the production of type 2 cytokines, which characterize allergic diseases . METHODS: PBMCs from patients allergic to house dust mite versus those from healthy donors were stimulated for 48 hours with the related Dermatophagoides pteronyssinus allergen or with a staphylococcal superantigen . The effect of LAB preincubation was assessed by measuring the type 2 cytokine production by means of specific ELISA . RESULTS: The tested gram-positive LAB were shown to inhibit the secretion of T(H)2 cytokines (IL-4 and IL-5) . This effect was dose dependent and was observed irrespective of the LAB strain used . No significant inhibition was induced by the control, gram-negative Escherichia coli TG1 . Interestingly, LAB reduced the T(H)2 cytokine production from allergic PBMCs specifically restimulated with the related allergen . The inhibition mechanism was shown to be dependent on antigen-presenting cells (ie, monocytes) and on the involvement of IL-12 and IFN-gamma . CONCLUSION: The tested LAB strains were demonstrated to exhibit an anti-T(H)2 activity, and thus different strains of this family might be useful in the prevention of allergic diseases.

J Immunol, 2002 Oct 15, 169(8), 4183 - 9
Generation ex vivo of TGF-beta-producing regulatory T cells from CD4+CD25- precursors; Zheng SG et al.; Previously we reported that TGF-beta has an important role in the generation and expansion of human "professional" CD4(+)CD25(+) regulatory T cells in the periphery that have a cytokine-independent mechanism of action . In this study we used low-dose staphylococcal enterotoxin to induce T cell-dependent Ab production . We report that TGF-beta induces activated CD4(+)CD25(-) T cells to become Th3 suppressor cells . While stimulating CD4(+) cells with TGF-beta modestly increased expression of CD25 and intracellular CTLA-4 in primary cultures, upon secondary stimulation without TGF-beta the total number and those expressing these markers dramatically increased . This expansion was due to both increased proliferation and protection of these cells from activation-induced apoptosis . Moreover, adding as few as 1% of these TGF-beta-primed CD4(+) T cells to fresh CD4(+) cells and B cells markedly suppressed IgG production . The inhibitory effect was mediated by TGF-beta and was also partially contact dependent . Increased TGF-beta production was associated with a decreased production of IFN-gamma and IL-10 . Depletion studies revealed that the precursors of these TGF-beta-producing CD4(+) suppressor cells were CD25 negative . These studies provide evidence that CD4(+)CD25(+) regulatory cells in human blood consist of at least two subsets that have TGF-beta-dependent and independent mechanisms of action . TGF-beta has an essential role in the generation of both of these T suppressor cell subsets from peripheral T cells . The ability to induce CD4(+) and CD8(+) cells to become regulatory cells ex vivo has the potential to be useful in the treatment of autoimmune diseases and to prevent transplant rejection.

J Perinatol, 2002 Oct-Nov, 22(7), 547 - 9
A comparison of two versus one blood culture in the diagnosis and treatment of coagulase-negative staphylococcus in the neonatal intensive care unit; Struthers S et al.; OBJECTIVES: This study compares two versus one blood culture in the diagnosis and treatment of coagulase-negative staphylococcus (CONS) in babies with suspected sepsis . STUDY DESIGN: The study was performed at British Columbia's Children's Hospital Neonatal Intensive Care Unit between March 1999 to March 2000 . One hundred pairs of cultures were drawn from two percutaneous sites from babies more than 48 hours old at the time of a sepsis screen . CONS cultured from both sites was regarded as evidence of infection . Positive culture from only one of the two sites was regarded as contaminant . The difference in rates of diagnosed CONS infection and reduction in antibiotic usage when using two versus one blood culture was calculated . RESULTS: In 5% of babies, cultures from a second site did not substantiate the diagnosis of CONS when compared to the result from a single culture . The resultant reduction in antibiotic use was 8.2% . CONCLUSIONS: Two blood cultures reduce the number of children diagnosed with CONS infection and reduce antibiotic usage.

Curr Allergy Asthma Rep, 2002 Nov, 2(6), 500 - 6
The role of infection in chronic rhinosinusitis; Bhattacharyya N; Chronic rhinosinusitis is a common condition, yet little is understood about its pathogenesis . Chronic infection traditionally has been considered a significant factor in the etiology and manifestations of chronic rhinosinusitis . Bacteria can be recovered in most cases of chronic rhinosinusitis, most commonly consisting of Staphylococcus species, anaerobes, and in some cases, gram-negative bacteria . Increasing trends toward bacterial resistance have been identified in chronic rhinosinusitis . Recently, a potential role for fungal infection has emerged . A knowledge of the microbiology of chronic rhinosinusitis will help guide treatment, but more research is required to understand further the exact role of infection in the pathophysiology of chronic rhinosinusitis.

Echocardiography, 2002 Aug, 19(6), 509 - 11
Left-sided endocarditis in intravenous drug users: a case report and review of the literature; Seghatol F et al.; We report a case of staphylococcus endocarditis of the mitral and aortic valves in an intravenous drug user (IVDU) complicated by abscess of the aortic root and aorto-left atrial fistula . Interestingly, the tricuspid valve was free of vegetation . Infective endocarditis in IVDUs more commonly involves right-sided valves; left-sided endocarditis is rare, indicates severe disease, or is a postmortem finding . This case illustrates the need for considering left-sided valve endocarditis in IVDU with septicemia, even if the tricuspid valve shows no evidence of vegetation.

Plant Cell, 1990 Nov, 2(11), 1091 - 1106
In Vitro Processing of Aleurain, a Barley Vacuolar Thiol Protease; Holwerda BC et al.; Aleurain, originally described from its cDNA as a thiol protease {Rogers, J.C., Dean, D., and Heck, G.R . (1985) . Proc . Natl . Acad . Sci . USA 82, 6512-6516}, is characterized here as a glycoprotein that is targeted to a distinct vacuolar compartment in aleurone cells . Monospecific antibodies to a bacterial trpE-aleurain fusion protein were used to show that aleurain is made as a 42-kilodalton (kD) proenzyme (proaleurain) that is proteolytically processed in a post-Golgi compartment in two steps to form a 32-kD protein . The first processing step is the discrete loss of 9 kD from proaleurain to yield a 33-kD intermediate that is further processed by the gradual loss of 1 kD resulting in mature 32-kD aleurain . Using proaleurain secreted from Xenopus oocytes as a substrate, we established an in vitro system using aleurone cell extracts that correctly processes proaleurain to a stable protein that is indistinguishable from native barley aleurain as judged by partial digestion with staphylococcal V8 protease . Proaleurain is not capable of self-cleavage in the absence of aleurone cell extracts and mature aleurain appears not to participate in processing in vitro.

Mol Microbiol, 2002 Sep, 45(6), 1485 - 98
Controlling pore assembly of staphylococcal gamma-haemolysin by low temperature and by disulphide bond formation in double-cysteine LukF mutants; Nguyen VT et al.; Staphylococcal LukF and Hlg2 are water-soluble monomers of gamma-haemolysin that assemble into oligomeric pores on the erythrocyte membranes . Here, we have created double-cysteine LukF mutants, in which single disulphide bonds connect either the prestem domain and the cap domain (V12C-T136C, Cap-Stem), or two beta-strands within the prestem domain (T117C-T136C, Stem-Stem) to control pore assembly of gamma-haemolysin at intermediate stages . The disulphide-trapped mutants were inactive in erythrocyte lysis, but gained full haemolytic activity if the disulphide bonds were reduced . The disulphide bonds blocked neither the membrane binding ability nor the intermediate prepore oligomerization, but efficiently inhibited the transition from prepores to pores . The prepores of Cap-Stem were dissociated into monomers in 1% SDS . In contrast, the prepores of Stem-Stem were stable in SDS and had ring-shaped structures similar to those of wild-type LukF, as observed by transmission electron microscopy . The transition of both mutants from prepores to pores could even be achieved by reducing disulphide bonds at low temperature (2 degrees C), whereas prepore oligomerization was effectively inhibited by low temperature . Finally, real-time transition of Stem-Stem from prepores to pores on ghost cells, visualized using a Ca2+-sensitive fluorescent indicator (Rhod2), was shown by the sequential appearance of fluorescence spots, indicating pore-opening events . Taken together, these data indicate that the prepores are legitimate intermediates during gamma-haemolysin pore assembly, and that conformational changes around residues 117 and 136 of the prestem domain are essential for pore formation, but not for membrane binding or prepore oligomerization . We propose a mechanism for gamma-haemolysin pore assembly based on the demonstrated intermediates.

Int Immunopharmacol, 2002 Jul, 2(8), 1143 - 53
Thalidomide can costimulate or suppress CD4+ cells' ability to incorporate {H3}-thymidine--dependence on the primary stimulant; Shannon EJ et al.; Thalidomide is a drug that can enhance mitogen- and antigen-stimulated cells' ability to synthesize IL-2 . To assess if thalidomide could concomitantly enhance the synthesis of IFN-gamma and incorporation of {H3}-thymidine, peripheral blood mononuclear cells (PBMC) were incubated in the presence or absence of thalidomide and staphylococcal enterotoxin A (SEA), anti-CD3, Con-A or PHA . After 18 h, the cultures were sampled for IL-2 . At the termination of the 3-day cultures, they were assayed for IFN-gamma and incorporation of {H3}-thymidine . Regardless of the mitogen used to stimulate the PBMC, the thalidomide-treated PBMC produced more IL-2 than controls . Thalidomide enhanced IFN-gamma synthesis in the Con-A and anti-CD3-stimulated PBMC . It suppressed the ability of SEA and PHA-stimulated PBMC to incorporate {H3}-thymidine, whereas it enhanced incorporation of {H3}-thymidine in PBMCs stimulated with anti-CD3 . When the PBMC were enriched for CD4+ or CD8+ cells, the SEA- and anti-CD3-stimulated CD4+ cells responded far better than the CD8+ cells in the synthesis of IL-2 and incorporation of {H3}-thymidine . In the thalidomide-treated SEA-stimulated CD4+ and CD8+ cells, thalidomide acted as a costimulant to enhance the synthesis of IL-2 . In the anti-CD3-stimulated thalidomide-treated cultures of PBMC enriched for CD4+ cells, thalidomide acted as a costimulant to enhance the incorporation {H3}-thymidine . Thalidomide cooperated with all of the mitogens to enhance T-cell synthesis of IL-2 . However, depending on the stimulant, thalidomide could suppress or enhance PBMC incorporation of {H3}-thymidine . The SEA-stimulated cells targeted by thalidomide to suppress incorporation of {H3}-thymidine were CD4+ . CD4+ cells stimulated with anti-CD3 were enhanced by thalidomide in their ability to synthesize IL-2 and to incorporate {H3}-thymidine . Increased production of IL-2 by activated T cells may be a mechanism through which thalidomide exerts its immunomodulatory effects.

J Obstet Gynaecol India, 1978 Jun, 28(3), 429 - 35
Pelvic inflammation (a study of 800 cases); Shah HN et al.; PIP: 800 cases of pelvic inflammation (PI) admitted to the Civic Hospital, Ahmedabad, between October 1967 and August 1974 were studied . 772 (96.5) of these cases involved chronic PI (420 of doubtful etiology, 122 postpartum, 76 postabortal, 46 tuberculous, 33 gonococcal, and 75 postinstrumental) . A further 28 cases (3.5%) represented acute PI (19 of doubtful etiology, 5 postinstrumental, and 4 postabortal) . 87% of cases were 20-35 years of age . The most frequent clinical symptom was some form of menstrual disturbance (polymenorrhea, 14.87%; dysmenorrhea, 13.5%; scanty period, 11.13%; menorrhagia, 13.37%; and irregular, profuse period, 14%) . 48% of the women complained of acute or chronic lower abdominal pain, and 31.1% experienced leukorrhea or purulent vaginal discharge . Sterility was found in 25% of cases . 50% of women with acute PI, 37.6% of those with chronic PI, and 6.52% of those with tubercular PI reported regular menstrual cycles . The most common organism isolated was Escherichia coli (38.45% of cases) . Other organisms detected included staphylococcus (10.75%) and gonococci (8.61%) . No organism was isolated in 33.78% of cases . The response to treatment with various antibiotics and cortisone was considered good in 28% of cases, fair in 37%, and poor or no response in 36% . Patients given a combined therapy of chloromycetin and corticosteroids showed early recovery and complete resolution of inflammatory masses . Follow-up study revealed that menstrual disorders were cured in 57% of cases and abdominal pain was relieved in 53% .

Dev Sante, 1997 Aug, (130), 5 - 10
{Infectious respiratory emergencies in children}; Bottineau MC et al.; PIP: Respiratory infections that may prove fatal for young children without swift and energetic emergency treatment are described . Respiratory infections, along with diarrhea, continue to be a major cause of infant mortality . Six characteristic conditions are addressed: three pulmonary infections and 3 tracheo-laryngeal infections . Each condition is described; its clinical signs are outlined; the course of the disease, possible complications, and signs of gravity are discussed; and treatments are recommended, with second choices indicated . The three pulmonary infections described are pleuro-pulmonary staphylococcus, severe whooping cough, and acute tuberculosis . The tracheo-laryngeal infections are epiglottitis, staphylococcal laryngeo-tracheitis, and acute tonsillitis .

Immunol Lett, 2002 Nov 1, 84(2), 97 - 101
Different mechanisms are utilized by HIV-1 Nef and staphylococcal enterotoxin A to control and regulate interleukin-10 production; Tangsinmankong N et al.; Interleukin-10 (IL-10) plays an important immunopathogenic role in immunologic diseases, especially in HIV infection and atopic dermatitis . The control and regulatory mechanisms of IL-10 production have not been described in these diseases . Recently, we demonstrated that HIV-1 Nef induces IL-10 production in monocytes and that staphylococcal enterotoxin A (SEA) induces IL-10 production in T-lymphocytes . Here we show that Nef-induced IL-10 production and mRNA expression are strongly blocked by rapamycin, but are not blocked by cyclosporin (CsA) or FK506 . Conversely, we show that CsA and FK506 completely inhibit SEA-induced IL-10 protein production and mRNA expression . The results of this study demonstrate that IL-10 production by Nef and SEA is controlled and regulated by different mechanisms.

Life Sci, 2002 May 3, 70(24), 2897 - 913
Effects of Shuanghuanglian and Qingkailing, two multi-components of traditional Chinese medicinal preparations, on human leukocyte function; Chen X et al.; Qingkailing (QKL) and Shuanghuanglian (SHHL) are two commonly used Chinese herbal preparations with reported antiinflammatory activity . The effects of these two preparations on the capacity of staphylococcal toxic shock syndrome toxin 1 (TSST-1) to stimulate the production of cytokines (IL-1beta, IL-6, TNF-alpha, IFN-gamma) and chemokines (MIP-1alpha, MIP-1beta and MCP-1) by peripheral blood mononuclear cell (PBMC) was tested . We also evaluated their effect on LPS-stimulated NF-kappaB transcriptional activity in a THP-1 cell line, and on human monocyte chemotactic response to chemoattractants . Non-cytotoxic concentrations of QKL (0.1 to approximately 2%) and SHHL (6 to approximately 120 microg) significantly inhibited production of cytokines and chemokines in a dose-dependent manner (P < 0.05) . Both, QKL at 1:100 and SHHL at 60 microg/ml, markedly inhibited RANTES, MIP-1alpha, SDF-1alpha and fMLP induced human monocyte migration (P < 0.05 or 0.01) . QKL (1%) did not inhibit monocyte chemotaxis induced by super-or sub-optimal concentrations of fMLP (10(-5), 10(-6) and 10(-10) M), but only inhibited chemotaxis induced by optimal concentrations of fMLP at 10(-7), 10(-8) and 10(-9) M . QKL (0.1% or 1%) and SHHL (6 or 60 microg/ml) markedly inhibited LPS-induced NF-kappaB activity in THP-1 cells . The results suggested that the pharmacological basis for the antiinflammatory effects of QKL and SHHL is the result of suppression of NF-kappaB regulated gene transcription, leading to suppressed production of proinflammatory cytokine and chemokine . Interference with leukocyte chemotaxis also contributes to the antiinflammatory and immunomodulating effects of these medicinals . Identification of the responsible components in these two herbal preparations may yield compounds suitable for structural modification into potent novel drugs.

Arch Dis Child, 2002 Oct, 87(4), 312 - 5
Clinical and radiographic spectrum of septic pulmonary embolism; Wong KS et al.; AIMS: To review the clinical presentation, radiographic findings, and outcome of therapy in children with septic pulmonary embolism . METHODS: Retrospective analysis of patients in a tertiary paediatric facility in northern Taiwan . RESULTS: Ten children were identified with septic pulmonary emboli in a four year retrospective chart review between 1998 and 2001 . Seven were immunocompetent, two were premature infants, one had beta thalassemia major . Seven had community acquired staphylococcal infections and bacteraemia, of which six were methicillin resistant Staphylococus aureus (MRSA) isolates . Five had soft tissue infections, two bone infections, one suppurative otitis media, one catheter related infection, and one unknown foci of infection . Multiple and bilateral nodular pulmonary parenchymal lesions were common on plain chest radiographs, but chest computed tomography scans showed the additional findings of a "vessel sign" and central cavitations, confirming the existence of septic pulmonary embolism . CONCLUSIONS: Community acquired MRSA infections occurred in seven patients with septic pulmonary embolism but without predisposing high risk factors . Critically ill children with skin, soft tissue, or bone infections, when associated with septic pulmonary embolism in an area with a high rate of MRSA, should be empirically treated with glycopeptides (such as vancomycin or teicoplanin) before susceptibility results are known, in order to minimise morbidity and avoid mortality.

Proc Natl Acad Sci U S A, 2002 Oct 1, 99(20), 12612 - 6 Epub 2002 Sep 18.
A method for the generation of combinatorial antibody libraries using pIX phage display; Gao C et al.; For more than a decade, phage displayed combinatorial antibody libraries have been used to generate and select a wide variety of antibodies . We previously reported that the phage coat proteins pVII and pIX could be used to display the heterodimeric structure of the antibody Fv region . Herein, aspects of this technology were invoked and extended to construct a large, human single-chain Fv (scFv) library of 4.5 x 10(9) members displayed on pIX of filamentous bacteriophage . Furthermore, the diversity, quality, and utility of the library were demonstrated by the selection of scFv clones against six different protein antigens . Notably, more than 90% of the selected clones showed positive binding for their respective antigens after as few as three rounds of panning . Analyzed scFvs were also found to be of high affinity . For example, kinetic analysis (BIAcore) revealed that scFvs against staphylococcal enterotoxin B and cholera toxin B subunit had a nanomolar and subnanomolar dissociation constant, respectively, affording affinities comparable to, or exceeding that, of mAbs obtained from immunization . High specificity was also attained, not only between very distinct proteins, but also in the case of the Ricinus communis ("ricin") agglutinins (RCA(60) and RCA(120)), despite >80% sequence homology between the two . The results suggested that the performance of pIX-display libraries can potentially exceed that of the pIII-display format and make it ideally suited for panning a wide variety of target antigens.

N Engl J Med, 2002 Sep 19, 347(12), 869 - 77
Environmental exposure to endotoxin and its relation to asthma in school-age children; Braun-Fahrlander C et al.; BACKGROUND: In early life, the innate immune system can recognize both viable and nonviable parts of microorganisms . Immune activation may direct the immune response, thus conferring tolerance to allergens such as animal dander or tree and grass pollen . METHODS: Parents of children who were 6 to 13 years of age and were living in rural areas of Germany, Austria, or Switzerland where there were both farming and nonfarming households completed a standardized questionnaire on asthma and hay fever . Blood samples were obtained from the children and tested for atopic sensitization; peripheral-blood leukocytes were also harvested from the samples for testing . The levels of endotoxin in the bedding used by these children were examined in relation to clinical findings and to the cytokine-production profiles of peripheral-blood leukocytes that had been stimulated with lipopolysaccharide and staphylococcal enterotoxin B . Complete data were available for 812 children . RESULTS: Endotoxin levels in samples of dust from the child's mattress were inversely related to the occurrence of hay fever, atopic asthma, and atopic sensitization . Nonatopic wheeze was not significantly associated with the endotoxin level . Cytokine production by leukocytes (production of tumor necrosis factor alpha, interferon-gamma, interleukin-10, and interleukin-12) was inversely related to the endotoxin level in the bedding, indicating a marked down-regulation of immune responses in exposed children . CONCLUSIONS: A subject's environmental exposure to endotoxin may have a crucial role in the development of tolerance to ubiquitous allergens found in natural environments .

Radiat Res, 2002 Oct, 158(4), 516 - 22
Scattering phenomena effects on growth of 308-nm laser-irradiated bacteria in suspension; Pompa PP et al.; In this study we analyzed the effect of 308-nm laser exposure on recovery of irradiated Staphylococcus epidermidis held in liquid after irradiation and before plating . Coexistence of bacterial growth inhibition and stimulation phenomena was observed . Under certain conditions, bacterial recovery was about fivefold higher in irradiated samples than in the controls . The available evidence suggests that the growth inhibition was due to the bactericidal activity of the 308-nm wavelength light, whereas the growth stimulation effect was associated with broadband radiation generated by scattering phenomena in the bacterial suspensions . Spectroscopic investigations revealed that the nutrient broth plays a decisive role in the scattering of laser radiation within the suspension.

EMBO J, 2002 Sep 16, 21(18), 4950 - 8
Identification of p100 as a coactivator for STAT6 that bridges STAT6 with RNA polymerase II; Yang J et al.; STAT6 is a central mediator of IL-4-induced gene responses . STAT6-mediated transcription is depend ent on the C-terminal transcription activation domain (TAD), but the mechanisms by which STAT6 activates transcription are poorly understood . Here, we have identified the staphylococcal nuclease (SN)-like domain and tudor domain containing protein p100 as a STAT6 TAD interacting protein . p100 was originally characterized as a transcriptional coactivator for Epstein-Barr virus nuclear antigen 2 . STAT6 interacted with p100 in vitro and in vivo . The interaction was mediated by the TAD domain of STAT6 and the SN-like domain of p100 . p100 did not affect the immediate activation events of STAT6, but enhanced STAT6-mediated transcriptional activation and the IL-4-induced Igepsilon gene transcription in human B-cell line . Finally, p100 associated with the large subunit of RNA polymerase II and was mediating interaction between STAT6 and RNA polymerase II . These findings identify p100 as a novel coactivator for STAT6 and suggest that p100 functions as a bridging factor between STAT6 and the basal transcription machinery.

Rev Med Liege, 2002 Jul, 57(7), 459 - 62
{Homograft mitral valve replacement}; Radermecker MA et al.; The total homograft replacement of the left AV valve and subvalvular apparatus constitutes a delicate operation, which may prove beneficial in young patients with extensive bacterial endocarditis . The case of a staphylococcal endocarditis in a young drug addicted patient, operated three years previously of mitral valve repair for the same pathology, is presented . In addition to the complete excision of infected tissues and valvular substitution with biological material, this technique has the advantage of avoiding long term anticoagulation . The history, technical key points, and current indications of mitral homograft in the surgery of the left atrio-ventricular valves are reviewed.

J Endotoxin Res, 2002, 8(4), 253 - 61
Staphylococcal enterotoxin B induces hepatic injury and lethal shock in endotoxin-resistant C3H/HeJ mice despite a deficient macrophage response; Yasuda S et al.; Bacterial toxins, including endotoxin/LPS as well as superantigens, are major causative agents of multi-organ failure associated with sepsis and liver disease . However, the precise mechanisms initiating cell activation by the toxins have not been clarified . We compared lethal shock and cytokine production in response to LPS with responses to the superantigen staphylococcal enterotoxin B (SEB) in both LPS-responsive C3H/HeN mice and LPS-hyporesponsive C3H/HeJ mice treated with D-galactosamine (GalN) . LPS was not lethal and did not induce production of TNF-alpha in C3H/HeJ mice . In contrast, SEB produced lethal shock associated with liver failure and induced cytokines such as TNF-alpha, IFN-gamma, and IL-2 in both C3H/HeN and C3H/HeJ mice . Peritoneal macrophages from C3H/HeJ mice did not produce TNF-alpha in vitro in response to SEB or LPS . However, no significant difference was observed in production of TNF-alpha in response to stimulation in vitro by SEB between C3H/HeN and C3H/HeJ splenic lymphocytes . We have demonstrated that SEB causes lethal toxicity associated with liver injury in LPS-hyporesponsive C3H/HeJ mice and that as the underlying mechanism, the normal T-cell function in these mice still maintained the sensitivity to SEB since the genetic defect of C3H/HeJ mice unresponsive to LPS and SEB is restricted in macrophages/monocytes and does not extend to T cells.

Perit Dial Int, 2002 May-Jun, 22(3), 335 - 8
Challenging the current treatment paradigm for methicillin-resistant Staphylococcus epidermidis peritonitis in peritoneal dialysis patients; Ariano RE et al.; OBJECTIVES: To analyze clinical outcomes of Staphylococcus epidermidis peritoneal dialysis peritonitis before and after an interventional switch from a vancomycin/ tobramycin to a cefazolin/tobramycin regimen for empiric treatment . To examine risk factors associated with clinical failure . DESIGN: A retrospective study . SETTING: A peritoneal dialysis program within a university-affiliated tertiary-care hospital . PATIENTS: 93 episodes of S . epidermidis peritonitis over a 6-year period . INTERVENTIONS: Clinical responses were compared between treatments using chi-square or Fisher's exact test . Univariate and multivariate analyses were used to identify significant risk factors for clinical failure . MEASUREMENTS AND MAIN RESULTS: There was no difference in the overall response rates observed with vancomycin (40/49; 81.6%) and cefazolin (23/29; 79.3%) regimens for episodes of S . epidermidis peritonitis . Furthermore, the presence of methicillin resistance in 63 of 93 cases (67.7%) had no influence on clinical outcome, with response rates of 83.9% (26/31) and 82.4% (14/17) for empiric vancomycin and cefazolin regimens, respectively . Tobramycin therapy of less than 2 days was an independent risk factor for clinical failure in multivariate logistic regression analysis (odds ratio 4.44, 95% confidence interval 1.28 - 15.48; p = 0.02) . CONCLUSIONS: Empiric treatment with intraperitoneal cefazolin was as effective as vancomycin for S . epiderimidis peritonitis despite a high prevalence of methicillin resistance . Tobramycin therapy of less than 2 days was strongly associated with treatment failure.

J Mol Med, 2002 Sep, 80(9), 585 - 94 Epub 2002 Jun 04.
Adenovirus-mediated intratumoral lymphotactin gene transfer potentiates the antibody-targeted superantigen therapy of cancer; Wang Q et al.; Bacterial superantigens are extremely potent activators of murine and human T lymphocytes . To engineer superantigens for cancer immunotherapy, staphylococcal enterotoxin A (SEA) was genetically fused to the Fab region of the human colon carcinoma-reactive monoclonal antibody (mAb) C215 . Fusion protein C215Fab-SEA can trigger cytotoxic T cells against C215 antigen positive tumor cells and induce tumor-suppressive cytokines . However, the antitumor effect of C215Fab-SEA is often not satisfactory because of T cell deletion after activation and failure to induce potent CTL activity after repeated administration . Lymphotactin (Lptn) is a potent chemoattractant for T cells and NK cells . To improve the therapeutic efficacy of fusion protein C215Fab-SEA we investigated in this study the antitumor responses elicited by combination of C215Fab-SEA and adenovirus-mediated intratumoral Lptn gene transfer in the preestablished C215 antigen expressing B16 melanoma murine model . More significant inhibition of tumor growth and prolonged survival time were observed in tumor-bearing mice that received combined therapy of C215Fab-SEA and Ad-Lptn than those of mice treated with C215Fab-SEA or Ad-Lptn alone . The highest CTL activity of tumor-bearing mice was induced after combined therapy . Intratumoral coadministration of C215Fab-SEA and Ad-Lptn augmented splenic NK activity of tumor-bearing mice most markedly . Our data demonstrate that the in vivo antitumor effect of C215Fab-SEA immunotherapy is potentiated significantly by combination with intratumoral Lptn gene transfer through more efficient induction of specific and nonspecific antitumor immune responses.

Biosci Biotechnol Biochem, 2002 Jul, 66(7), 1601 - 4
A highly sensitive assay for proteases using staphylococcal protein A fused with enhanced green fluorescent protein; Fujino H et al.; Enhanced green fluorescent protein (EGFP) was fused with staphylococcal protein A (SpA) and used as a substrate for proteases . An SpA-EGFP assay was done in three steps: (i) digestion of SpA-EGFP by proteases, (ii) addition of rabbit IgG immobilized on Sepharose beads, and (iii) measurement of the fluorescence intensity of supernatant . The assay was sensitive enough to measure picogram levels of trypsin and chymotrypsin, and may be applicable to various other proteases as one of the most sensitive methods.

Vet Microbiol, 2002 Oct 2, 89(1), 83 - 94
Trends in antimicrobial susceptibility in relation to antimicrobial usage and presence of resistance genes in Staphylococcus hyicus isolated from exudative epidermitis in pigs; Aarestrup FM et al.; From 1996 to 2001 a total of 467 Staphylococcus hyicus isolates from exudative epidermitis (EE) in pigs in Denmark were examined for susceptibility to 13 different antimicrobial agents . The presence of selected genes encoding macrolide (erm(A), erm(B) and erm(C)), penicillin (blaZ), streptogramin (vat, vga, vga(B), vat(B), vat(D) and vat(E)), streptomycin (aadE) and tetracycline resistance (tet(K), tet(L), tet(M) and tet(O)) were determined in selected isolates.The occurrence of erythromycin resistance increased from 33% in 1996 to a maximum of 62% in 1997 and decreased to 26% in 2001 . Resistance to sulphametazole increased from 17% in 1996 to 30% in 1998 but has since decreased to 4% in 2001 . Resistance to trimethoprim increased to 51% in 1997 and decreased to 21% in 2001 . Resistance to tetracycline (21-31%) remained relatively constant during 1996-2000, but increased to 47% in 2001 . Resistance to penicillin (54-75%) streptomycin (33-53%) and tetracycline (21-47%) remained relatively constant over the time investigated.All 48 penicillin resistant isolates examined contained the blaZ gene and 40 (85%) of the streptomycin resistant isolates the aadE gene . It was not possible to detect any streptogramin resistance gene in four streptogramin resistant isolates . Of the 55 erythromycin resistant isolates examined, five contained erm(A), 13 erm(B), 35 erm(C) and two both erm(A) and erm(C) . The presence of erm(B) was confirmed by hybridization to plasmid profiles in all 13 PCR-positive isolates . Of 52 tetracycline resistant isolates examined, two contained tet(L), 38 tet(K) and 12 both tet(K) and tet(L).

Am J Rhinol, 2002 Jul-Aug, 16(4), 181 - 6; discussion 186
The comparative risks of bacterial contamination between a venturi atomizer and a positive displacement atomizer; Wolfe TR et al.; INTRODUCTION: This laboratory study determined the incidence of internal contamination of Venturi principle atomizers and positive displacement atomizers exposed to high external concentrations of Staphylococcal aureus (Staph) . METHODS: Atomizer device nozzle tips were immersed into a Staph solution and I ml of spray was atomized via compressed wall air (Venturi) or hydraulic pump (positive displacement) . The Venturi nozzle was then wiped with 70% isopropyl alcohol while the disposable positive displacement nozzle was replaced . After 30 minutes, 1 ml of atomized fluid was collected and cultured and the process was repeated . After sixteen uses the fluid remaining in the bottles was cultured . The Venturi atomizer also was subjected to a single use trial to determine the location of device contamination . RESULTS: Venturi atomizers sprays grew Staph in every case (144/144), while positive displacement atomizer sprays never grew contaminants (0/144; p < 0.0001) . At the end of 16 uses, 7/9 of Venturi atomizers had Staph within their medication reservoirs while none (0/9; p = 0.002) existed in the positive displacement atomizers . After a single use of the Venturi atomizer, the medication reservoir, the air lumen and the medication lumen of the nozzle were all contaminated with Staph . CONCLUSIONS: External bacterial contamination of the atomizer nozzle tip results in internal bacterial contamination of Venturi devices in as little as one use but not of positive displacement devices . These results warrant further investigation to determine whether a risk of cross-contamination exists in a clinical setting.

Biochemistry, 2002 Sep 17, 41(37), 11118 - 25
Enhanced strand invasion by peptide nucleic acid-peptide conjugates; Kaihatsu K et al.; Efficient and selective recognition of DNA by proteins is due to sequence-specific interactions with a target site and nonselective electrostatic interactions that promote the target's rapid location . If synthetic molecules could mimic these functions, they would render a wide range of chromosome sequences accessible to rationally designed probes . Here we describe conjugates between bispeptide nucleic acids (bisPNAs) designed to specifically recognize duplex DNA and peptides that have been designed to promote rapid sequence recognition . Peptide design was based on the surface of staphylococcal nuclease, a cationic DNA binding protein with low sequence selectivity . We observe that attachment of the designed peptide increases rates of strand invasion by 100-fold relative to unmodified bisPNA . The peptide can contain D-amino acids, increasing the likelihood that it will be stable in cell extract and inside cells . Binding of the conjugate containing the D-amino acid peptide occurred over a broad range of experimental conditions and was sensitive to a single mismatch . Strand invasion was efficient at neutral to basic pH, a wide range of temperatures (0-65 degrees C), and in the presence of up to 7 mM Mg(2+) and 100 mM Na(+) or K(+) . Our data suggest that attachment of peptides that mimic cationic protein surfaces to PNAs can afford conjugates that mimic the rapid and selective binding that characterizes native DNA binding proteins . Rapid strand invasion over a wide range of experimental conditions should further expand the utility of strand invasion by PNAs.

Semin Arthritis Rheum, 2002 Aug, 32(1), 3 - 9
Vasculitis secondary to staphylococcal Protein A immunoadsorption (Prosorba column) treatment in rheumatoid arthritis; Deodhar A et al.; OBJECTIVE: Vasculitis is a rare complication of immunoadsorption treatment with staphylococcal Protein A (Prosorba column) . The prevalence, clinical characteristics, pathophysiology, treatment, and outcome of vasculitis secondary to immunoadsorption treatment is not known . METHODS: The authors describe a 57-year-old woman with rheumatoid arthritis (RA) resistant to methotrexate and etanercept, who developed severe leukocytoclastic vasculitis after the 9th weekly treatment with Prosorba column . She developed rapidly progressive crescentic glomerulonephritis and required treatment with cyclophosphamide and high-dose prednisone . Subsequently, renal function stabilized and RA remitted . Through the literature search and by reviewing information submitted to Cypress Biosciences Inc (manufacturer of Prosorba columns), available world literature on vasculitis secondary to Prosorba column treatment was compiled . RESULTS: Immune complex deposition of staphylococcal Protein A (SPA)/SPA antibodies in the glomeruli precipitated the renal disease in our patient . Twenty cases of vasculitis (calculated prevalence, 1 per 400), 5 with internal organ involvement, have been reported in patients treated with Prosorba column for thrombocytopenic purpura . Seven RA patients treated with Prosorba column developed vasculitis (prevalence, 7 per 400), 3 with internal organ involvement . CONCLUSIONS: Vasculitis secondary to staphylococcal Protein A immunoadsorption therapy occurs rarely and appears to be related to development of SPA/SPA antibody immune complexes . Rheumatologists should be aware of this potentially serious complication of the Prosorba column treatment for RA .

J Immunol, 2002 Sep 15, 169(6), 2907 - 14
Superantigen enhancement of specific immunity: antibody production and signaling pathways; Torres BA et al.; Superantigens are microbial proteins that induce massive activation, proliferation, and cytokine production by CD4+ T cells via specific Vbeta elements on the TCR . In this study we examine superantigen enhancement of Ag-specific CD4+ T cell activity for humoral B cell responses to T-dependent Ags BSA and HIV gp120 envelope, type I T-independent Ag LPS, and type II T-independent Ag pneumococcal polysaccharides . Injection of BSA followed by a combination of superantigens staphylococcal enterotoxin A and staphylococcal enterotoxin B (SEB) 7 days later enhanced the anti-BSA Ab response in mice approximately 4-fold as compared with mice given BSA alone . The anti-gp120 response was enhanced approximately 3-fold by superantigens . The type II T-independent Ag pneumococcal polysaccharide response was enhanced approximately 2.3-fold by superantigens, whereas no effect was observed on the response to the type I T-independent Ag LPS . The superantigen effect was completely blocked by the CD4+ T cell inhibitory cytokine IL-10 . SEB-stimulated human CD4+ T cells were examined to determine the role of the mitogen-activated protein (MAP) kinase signal transduction pathway in superantigen activation of T cells . Inhibitors of the mitogen pathway of MAP kinase blocked SEB-induced proliferation and IFN-gamma production, while an inhibitor of the p38 stress pathway had no effect . Consistent with this, SEB activated extracellular signal-regulated kinase/MAP kinase as well as MAP kinase-interacting kinase, a kinase that phosphorylates eIF4E, which is an important component of the eukaryotic protein synthesis initiation complex . Both kinases were inhibited by IL-10 . Thus, superantigens enhance humoral immunity via Ag-specific CD4+ T cells involving the stress-independent pathway of MAP kinase.

J Infect, 2002 Aug, 45(2), 129 - 32
Treatment of Staphylococcus epidermidis ventriculo-peritoneal shunt infection with linezolid; Gill CJ et al.; Gram-positive bacterial meningitis frequently complicates ventriculo-peritoneal (VP) shunts used for hydrocephalus . Linezolid, an oxazolidinone, is active against Gram-positive cocci, and has excellent CSF penetration . We present a 22-year-old woman who was cured of a Staphylococcus epidermidis VP shunt infection via shunt removal and intravenous linezolid.

Eur J Vasc Endovasc Surg, 2002 Sep, 24(3), 230 - 4
Quinupristin/dalfopristin bonding in combination with intraperitoneal antibiotics prevent infection of knitted polyester graft material in a subcutaneous rat pouch model infected with resistant Staphylococcus epidermidis; Ghiselli R et al.; OBJECTIVE: to investigate the efficacy of quinupristin/dalfopristin in the prevention of prosthetic graft infection in a rat subcutaneous pouch model . METHODS: graft infections were established in the subcutaneous tissue of 140 male Wistar rats by implantation of Dacron prostheses followed by topical inoculation with Staphylococcus epidermidis with intermediate resistance to glycopeptides . The study included one group without contamination, one contaminated group without prophylaxis, one contaminated group that received 50mg/l quinupristin/dalfopristin-soaked graft, one contaminated group that received 10mg/kg intraperitoneal levofloxacin, one contaminated group that received 3mg/kg intraperitoneal doxycycline, and two contaminated groups that received 50mg/l quinupristin/dalfopristin-soaked plus 10mg/kg intraperitoneal levofloxacin or 3mg/kg intraperitoneal doxycycline . Each group included 20 animals . The grafts were removed after 7 days and evaluated by quantitative culture . RESULTS: quinupristin/dalfopristin showed a significantly higher efficacy than levofloxacin and doxycycline, even though quantitative graft cultures for rats that received only quinupristin/dalfopristin-soaked graft showed bacterial growth . Otherwise, the efficacy of levofloxacin was similar to that of doxycycline . Only the group treated with quinupristin/dalfopristin combined with levofloxacin or doxycycline showed no evidence of staphylococcal infection . CONCLUSIONS: quinupristin/dalfopristin as adjunctive topical antibiotic prophylaxis can be useful for the prevention of vascular graft infections caused by staphylococcal strains with high levels of resistance.

Hum Gene Ther, 2002 Aug 10, 13(12), 1427 - 39
Genetic modification of adenovirus 5 tropism by a novel class of ligands based on a three-helix bundle scaffold derived from staphylococcal protein A; Henning P et al.; The use of adenovirus (Ad) as an efficient and versatile vector for in vivo tumor therapy requires the modulation of its cellular tropism . We previously developed a method to genetically alter the tropism of Ad5 fibers by replacing the fiber knob domain by an extrinsic trimerization motif and a new cellular ligand . However, fibers carrying complex ligands such as single-chain antibody fragments did not assemble into functional pentons in vitro in the presence of penton base, and failed to be rescued into infectious virions because of their inability to fold correctly within the cytoplasm of Ad-infected cells . Here we show that the coding sequence for a disulfide bond-independent three-helix bundle scaffold Z, derived from domain B of Staphylococcal protein A and capable of binding to the Fc portion of immunoglobulin (Ig) G1, could be incorporated into modified knobless Ad fiber gene constructs with seven shaft repeats . These fiber gene constructs could be rescued into viable virions that were demonstrated to enter 293 cells engineered for IgG Fc surface expression but not unmodified 293 cells, via a mechanism that could be specifically blocked with soluble Fc target protein . However, the tropism modified viruses showed a slightly impaired cellular entry and a lower infectivity than wildtype (WT) virus . In addition, we generated recombinant fibers containing an IgA binding Affibody ligand, derived from combinatorial specificity-engineering of the Z domain scaffold . Such fiber constructs also showed the expected target specific binding, indicating that the affibody protein class is ideally suited for genetic engineering of Ad tropism.

Proteins, 2002 Nov 1, 49(2), 255 - 65
Role of C-terminal region of Staphylococcal nuclease for foldability, stability, and activity; Hirano S et al.; The role of the C-terminal region of Staphylococcal nuclease (SNase) was examined by deletion mutation . Deletions up to eight residues do not affect the structure and function . The structure and enzymatic activity were partially lost by deleting Ser141-Asn149 (Delta141-149), and deletion of Trp140-Asn149 (Delta140-149) resulted in further loss of structure and activity . A 13-residue deletion showed the same effect as the 10-residue deletion . Both Ser141Gln and Ser141Ala mutations for an eight-residue deletion mutant did not alter properties as well as Ser141A1a for full-length SNase . In contrast, Trp140Ala mutation for Delta141-149 shows the same effect as the deletion of Trp140 . Trp140Ala mutation for full-length SNase causes the loss of native structure . These observations indicate the significance of the 140th and the 141st residues . The side-chain of the 140th residue is required to be tryptophan; however, the backbone of the 141st residue is solely critical for foldability, but the side-chain information is not crucial . All of the mutants that take a non-native conformation show enzymatic activity and inhibitor-induced folding, suggesting that foldability is required for the activity .

Intensive Care Med, 2002 Sep, 28(9), 1324 - 31 Epub 2002 Aug 10.
Diagnostic accuracy of G-CSF, IL-8, and IL-1ra in critically ill children with suspected infection; Fischer JE et al.; OBJECTIVE: To elucidate the diagnostic accuracy of granulocyte colony-stimulating factor (G-CSF), interleukin-8 (IL-8), and interleukin-1 receptor antagonist (IL-1ra) in identifying patients with sepsis among critically ill pediatric patients with suspected infection . DESIGN AND SETTING: Nested case-control study in a multidisciplinary neonatal and pediatric intensive care unit (PICU) PATIENTS: PICU patients during a 12-month period with suspected infection, and plasma available from the time of clinical suspicion (254 episodes, 190 patients) . MEASUREMENTS AND RESULTS: Plasma levels of G-CSF, IL-8, and IL-1ra . Episodes classified on the basis of clinical and bacteriological findings into: culture-confirmed sepsis, probable sepsis, localized infection, viral infection, and no infection . Plasma levels were significantly higher in episodes of culture-confirmed sepsis than in episodes with ruled-out infection . The area under the receiver operating characteristic curve was higher for IL-8 and G-CSF than for IL-1ra . Combining IL-8 and G-CSF improved the diagnostic performance, particularly as to the detection of Gram-negative sepsis . Sensitivity was low (<50%) in detecting Staphylococcus epidermidis bacteremia or localized infections . CONCLUSIONS: In this heterogeneous population of critically ill children with suspected infection, a model combining plasma levels of IL-8 and G-CSF identified patients with sepsis . Negative results do not rule out S . epidermidis bacteremia or locally confined infectious processes . The model requires validation in an independent data-set.

Pediatr Pulmonol, 2002 Sep, 34(3), 242 - 7
Evidence-based medicine in cystic fibrosis: how should practice change?
Conway SP.
This critical review of current practice and research in some aspects of cystic fibrosis (CF) concludes that, despite recent success in prolonging life expectancy, there is still room for improvement in terms of earlier diagnosis and earlier intervention in some of its complications . Specifically, the body of evidence implies that neonatal diagnosis; early attention to nutrition, staphylococcal and pseudomonal infection, and diabetes mellitus; and early treatment with dornase alfa can all be expected to improve patients' quality and quantity of life .

Int Immunol, 2002 Sep, 14(9), 1015 - 26
Role of CD44 in activation-induced cell death: CD44-deficient mice exhibit enhanced T cell response to conventional and superantigens; McKallip RJ et al.; T cells upon activation are known to up-regulate CD44 expression . However, the precise function of CD44 on activated T cells is not clear . In this report, we demonstrate that signaling through CD44 plays an important role in activation-induced cell death (AICD) . CD44 knockout (KO) mice had an elevated in vivo primary and in vitro secondary response to challenge with conalbumin, anti-CD3 mAb and staphylococcal enterotoxin A (SEA), which correlated with reduced AICD when compared to CD44 wild-type mice . In addition, CD44 KO mice exhibited increased delayed-type hypersensitivity response to dinitrofluorobenzene . In a model examining in vitro AICD, splenocytes from CD44 KO mice showed resistance to TCR-mediated apoptosis when compared to splenocytes from CD44 wild-type mice . In addition, signaling through CD44 led to increased apoptosis in TCR-activated but not resting T cells from CD44 wild-type mice without affecting Fas expression . Injection of SEA into mice deficient in CD44 and Fas (CD44 KO/lpr) led to an increased primary response when compared to mice that expressed CD44 but not Fas (CD44 WT/lpr), suggesting that the enhanced response to SEA was dependent on CD44 but not Fas expression . Administration of anti-CD44 mAb into CD44 wild-type mice caused a significant decrease in antigen-specific T cell response . Together, these data implicate CD44 as an important regulator of AICD in T cells . Furthermore, targeting CD44 in vivo may constitute a novel approach to induce apoptosis in activated T cells, and therefore to treat autoimmune diseases, allograft rejection and graft versus host disease.

Blood, 2002 Sep 15, 100(6), 2216 - 24
Aberrant regulation of superantigen responses during T-cell reconstitution and graft-versus-host disease in immunodeficient mice; Spaner D et al.; Acute graft-versus-host disease (GVHD) after allogeneic stem cell transplantation is associated with impaired deletion and anergy of host-reactive T cells . To elucidate the immunoregulatory events that may contribute to such dysregulated T-cell responses in GVHD, we studied superantigen (SAg) responses after adoptive T-cell transfer into severe combined immunodeficient (SCID) mice . SAg responses are normally regulated by mechanisms involving deletion and anergy, with SAg-reactive T cells typically being deleted rapidly in vivo . In a SCID mouse model of GVHD, however, allogeneic host SAg-reactive T cells were not deleted rapidly, but rather persisted in increased numbers for several months . Moreover, depending on the timing of SAg stimulation and the numbers of T cells transferred, dysregulation (impaired deletion and anergy) of SAg responses could be demonstrated following the adoptive transfer of syngeneic T cells into SCID mice as well . Transgenic T-cell receptor-bearing KJ1-26.1(+) T cells were then used to determine the fate of weakly reactive T cells after adoptive transfer and SAg stimulation . When transferred alone, KJ1-26.1(+) T cells demonstrated impaired deletion and anergy . In the presence of more strongly staphylococcal enterotoxin B (SEB)-reactive T cells, however, KJ1-26.1(+) T cells were regulated normally, in a manner that could be prevented by inhibiting the effects of more strongly SEB-reactive cells or by increasing the level of activation of the KJ1-26.1(+) T cells themselves . We suggest that the control mechanisms that normally regulate strongly activated T cells in immunocompetent animals are lost following adoptive transfer into immunodeficient hosts, and that this impairment contributes to the development of GVHD.

Trends Immunol, 2002 Sep, 23(9), 461 - 3
Staphylococcal vaccines; Michie CA; Vaccination has proved relatively unsuccessful against the common mammalian commensal bacteria Staphylococcus, despite almost a century of experimentation . Recent progress in clinical trials, animal models and molecular laboratories offers hope that these organisms might be partially or wholly controlled by augmenting host responses.

Pediatr Med Chir, 2002 Jul-Aug, 24(4), 308 - 12
{Jacopo's chest (bad history of a pleural empyema) . A case report}; Celandroni A et al.; We present a case of pleural empyema, occurred in a healty 7 years boy . He was admitted to our hospital because of a lobare pneumonitis . The patient was administered with a 2 degrees generation Cefalosporine given intramuscularly and with Corticosteroid (1 mg/kg/die) . After an initial improvement of his clinical conditions, he got worse so that he underwent a TC scan which showed the presence of a left pleural empyema requiring the insertion of an intercostal tube drainage followed by an intervention of decortication . The boy had some evidence of a staphylococcal etiology such as the evolution in empyema itself, the augmentation of antistafilolisinic title found during the illness, and the typical finding of blebs on chest radiograph . As cultures from both blood and drainage liquid samples remained sterile, we were unable to demonstrate a clear bacterial etiology of the empyema . It remains doubtful if corticosteroid administration could contribute to the severity of the pneumonia evolution.

Biomaterials, 2002 Nov, 23(21), 4233 - 9
Detection of slime production by means of an optimised Congo red agar plate test based on a colourimetric scale in Staphylococcus epidermidis clinical isolates genotyped for ica locus; Arciola CR et al.; This investigation was conduced on a collection of 113 S . epidermidis strains isolated from biomaterial-associated infections . All strains were examined both for the presence of icaA and icaD genes responsible for slime synthesis by a PCR method and for the in vitro slime production ability by the Congo red agar (CRA) plate test . In the present study, the original CRA test was optimised adopting a six-colour reference scale for a fine classification of colonies colours . The six-colour tones of the scale were as follows: very black (vb), black (b), almost black (ab), which were considered as positive results, and bordeaux (brd), red (r), and very red (vr), interpreted as negative . 57.5% of all the strains were found to be icaA icaD-positive as well as slime-forming onto CRA, exhibiting the following colonies colours: vb (35.4%); b (15.9%); ab (6.2%) . The percentage of icaA icaD-negative strains was 42.5% and all of them were negative onto CRA: brd (19.5%), r (14.2%), vr (8.8%) . The comparison of colour classification with the information on ica genes confirmed the validity of the scale adopted, providing support to the criteria used for a correct interpretation of the colonies colour during the execution of the CRA test . Overall these results indicate a fine consistency between these two experimental methods and a good reliability of CRA plate test, especially when this is supported by a colourimetric scale.

Mikrobiol Z, 2002 Mar-Apr, 64(2), 59 - 64
{Correlation between Staphylococcus carriage, specific antibody-production and AB0-blood grouping in plasma donors}; Nemyrovs'ka LM et al.; Interaction peculiarities of three components of the immune human homeostasis-antigens of blood groups AB0, staphylococcus antigens and antistaphylococcus antibodies have been investigated . Donors (85) of antistaphylococcus plasma immunized by staphylococcus anatoxin have been investigated . It is found that the nasal staphylococcus carriage in donors depends on the level of specific and natural antibodies and on the coincidence between the staphylococcus antigen structure and the protein substance of the specific blood group factors.

Int Urogynecol J Pelvic Floor Dysfunct, 2002, 13(4), 232 - 4; discussion 235
The safety of reusing injectable collagen: a multicenter microbiological study; Culligan PJ et al.; We have previously reported pilot data regarding the safety of saving partially used syringes of a glutaraldehyde cross-linked collagen for use in subsequent treatment sessions with the same individual . That single institution study involved 56 partially used syringes cultured for aerobic bacteria . Only one weakly positive culture was detected among these 56 samples, which prompted us to carry out this expanded study involving multiple centers and different injection techniques . Samples were collected from four centers . Following periurethral injection in an office setting, 166 partially used syringes of glutaraldehyde cross-linked collagen were refrigerated for between 1 and 104 weeks (average 58) . Material from all 166 syringes was then cultured qualitatively and quantitatively for both aerobic and anaerobic organisms . Collagen from one syringe grew >100,000 colonies of Escherichia coli . All other cultures were negative . In the pilot study, one culture of 56 syringes was weakly positive for coagulase-negative staphylococcus . When the results from both studies were considered together, only two of 222 partially used syringes (0.9%) were contaminated . The background risk of local infection associated with periurethral collagen injection is approximately 0.29% . Using the statistical equation 'number needed to harm', we found that a clinician would have to reuse 111 syringes at a saving of $34,965 before he or she would cause a single local injection by so doing . Therefore, we feel that it may be cost-effective and safe to reinject material from a partially used syringe of glutaraldehyde cross-linked collagen during a subsequent treatment session on an individual.

J Bone Joint Surg Br, 2002 Jul, 84(5), 758 - 60
Development of resistant strains of Staphylococcus epidermidis on gentamicin-loaded bone cement in vivo; Thomes B et al.; We have compared the rates of infection and resistance in an animal model of an orthopaedic procedure which was contaminated with a low-dose inoculum of Staphylococcus epidermidis . We randomised 44 Sprague-Dawley rats to have bone cement implanted subcutaneously containing either gentamicin or saline (control) . The wound was inoculated with a dilute solution of gentamicin-sensitive Staphylococcus epidermidis . At two weeks the cement was retrieved and microbiologically tested . A lower overall rate of infection was seen in the gentamicin-loaded cement group, but there was a significantly higher rate of gentamicin-resistant infection in this group (Fisher's exact test, p < 0.01) . Antibiotic-impregnated cement has an optimum surface for colonisation and prolonged exposure to antibiotic allows mutational resistance to occur . Gentamicin-loaded cement may not be appropriate for revision surgery if it has been used already in previous surgery.

Biol Pharm Bull, 2002 Aug, 25(8), 981 - 5
Biological activity of 4-acetyltropolone, the minor component of Thujopsis dolabrata SIeb . et Zucc . hondai Mak; Morita Y et al.; 4-Acetyltropolone, a minor component of Thujopsis dolabrata SIEB . et Zucc . hondai MAKINO, showed antimicrobial activity against various microorganisms including wood-rotting fungi, a phytogrowth-inhibitory effect with chlorophyll biosynthesis inhibition, cytotoxic effect and inhibitory activity on metalloproteases . This compound had strong antifungal activity on Daedalea dickinsii IFO-4979 {minimum inhibitory concentration (MIC): 0.2 microg/ml} and Coriolus versicolor IFO-4940 (MIC: 0.39 microg/ml) . Its cytotoxic effect at 20.0/microg/ml on human stomach cancer KATO-III and Ehrich's ascites carcinoma was stronger than those of podophyllotoxin, vincristine and vinblastine, the anticancer agents isolated from higher plants and used clinically . This compound also had potent antibacterial activity against Staphylococcus epidermidis IFO-12993, its MIC being 1.56 microg/ml . However, other biological activities of 4-acetyltropolone were lower than those of hinokitiol which is the main component of this plant, suggesting that the contribution of the acetyl group at C-4 to biological activity is smaller than that of the isopropyl group at that position . The acute toxicity of 4-acetyltropolone (LD50: 335.2 mg/kg) to mice was much lower than that of hinokitiol (LD50: 191 mg/kg).

Antimicrob Agents Chemother, 2002 Sep, 46(9), 2885 - 8
Efficacy of quinupristin-dalfopristin in preventing vascular graft infection due to Staphylococcus epidermidis with intermediate resistance to glycopeptides; Giacometti A et al.; A rat model was used to investigate the efficacy of quinupristin-dalfopristin (Q-D) in the prevention of vascular prosthetic graft infection due to methicillin-resistant Staphylococcus epidermidis with intermediate resistance to glycopeptides . The in vitro activity of the compound was compared to that of vancomycin by MIC determination and time-kill study . Moreover, the efficacy of collagen-sealed Q-D-soaked Dacron was evaluated in a rat model of graft infection . Graft infections were established in the subcutaneous tissue of the backs of 120 adult male Wistar rats . The in vivo study included a control group, one contaminated group that did not receive any antibiotic prophylaxis, two contaminated groups that received grafts soaked with 10 and 100 micro g of Q-D per ml, respectively, and two contaminated groups that received grafts soaked with 10 and 100 micro g of vancomycin per ml, respectively . Rats that received Dacron grafts soaked with 100 micro g of Q-D per ml showed no evidence of infection (<10 CFU/ml) . In contrast, for rats that received Dacron grafts soaked with 10 micro g of Q-D per ml and Dacron grafts soaked with 10 or 100 micro g of vancomycin per ml, the quantitative graft cultures demonstrated 2.2 x 10(2) +/- 1.3 x 10(2), 2.2 x 10(6) +/- 1.9 x 10(5), and 5.6 x 10(2) +/- 0.3 x 10(2) CFU/ml, respectively . Taken together the results of the study demonstrate that the use of Dacron grafts soaked with Q-D can result in significant bacterial growth inhibition and show that this compound is potentially valuable for prevention of vascular prosthetic graft infection.

J Control Release, 2002 Aug 21, 82(2-3), 319 - 34
Sustained ex vivo skin antiseptic activity of chlorhexidine in poly(epsilon-caprolactone) nanocapsule encapsulated form and as a digluconate; Lboutounne H et al.; In this work, the sustained bactericidal activity of chlorhexidine base loaded poly(epsilon-caprolactone), PCL, nanocapsules against Staphylococcus epidermidis inoculated onto porcine ear skin was investigated . Drug loaded nanocapsules were prepared by the interfacial polymer deposition following solvent displacement method, then characterized by photon correlation spectroscopy, electrophoretic measurements, transmission and scanning electron microscopy . Antimicrobial activity of these colloidal carriers was evaluated (i) in vitro against eight strains of bacteria, and (ii) ex vivo against Staphylococcus epidermidis inoculated for 12 h onto porcine ear skin surface treated for 3 min either with 0.6% chlorhexidine base loaded or unloaded nanocapsules suspended in hydrogel, or 1% chlorhexidine digluconate aqueous solution . Chlorhexidine absorption into the stratum corneum (SC) was evaluated by the tape-stripping method . The results showed that chlorhexidine nanocapsules in aqueous suspension having a 200-300 nm size and a positive charge exhibited similar minimum inhibitory concentrations against several bacteria with chlorhexidine digluconate aqueous solution . Ex vivo, there was a significant reduction in the number of colony forming units (CFUs) from 3-min treated skin with chlorhexidine nanocapsule suspension (5 to <1 log(10)) compared to chlorhexidine digluconate solution (5 to 2.02 log(10)) after a 8-h artificial contamination . After a 12-h artificial contamination, both formulations failed to achieve a 5 log(10) reduction . Furthermore, from a 3-min treatment with an identical applied dose and a subsequent 12-h artificial contamination, a residual chlorhexidine concentration in the SC was found to be three-fold higher with chlorhexidine nanocapsule suspension than with chlorhexidine digluconate solution . Interestingly, nanocapsules were shown in porcine skin follicles . Consequently, a topical application of chlorhexidine base-loaded positively charged nanocapsules in an aqueous gel achieved a sustained release of bactericide against Staphylococcus epidermidis for at least 8 h . Enhancement of drug delivery by mediating a more direct and prolonged contact between the carrier and (i) bacteria, (ii) skin surface, and (iii) skin follicles was assumed.

Vet Dermatol, 2002 Aug, 13(4), 169 - 76
Adherence of Staphylococcus intermedius to canine corneocytes in vitro; Saijonmaa-Koulumies LE et al.; This study investigated the in vitro adherence of Staphylococcus intermedius to canine corneocytes, collected from a healthy dog using double-sided adhesive tape . Adherence was shown to depend on duration (P < 0.001) and temperature of incubation (P < 0.001) and the concentration of bacteria (P < 0.001) . Isolates of S . intermedius from lesions of pyoderma were not generally more adherent to healthy canine skin than were isolates from healthy dogs . Significant differences in adherence were demonstrated between individual isolates within both groups (P < 0.001) . The study suggests that among S . intermedius there is no correlation between virulence and adherence to canine corneocytes in vitro . The finding may be important for the potential use of avirulent variants of S . intermedius as antagonistic strains against canine pyoderma . However, more studies are needed to compare the adherence of the isolates to skin cells obtained from dogs with diseases predisposed to pyoderma.

J Neurosurg Anesthesiol, 2002 Jul, 14(3), 229 - 33
Toxic epidermal necrolysis after phenytoin usage in a brain trauma patient; Schummer W et al.; Toxic epidermal necrolysis is a drug-induced, rare, but life-threatening skin eruption . The main differential diagnoses are drug-induced erythema (hypersensitivity syndrome), acute graft-versus-host disease, staphylococcal scalded skin syndrome, and toxic shock syndrome . Because the therapy for toxic epidermal necrolysis and acute graft-versus-host disease differs largely from the others, it is necessary to make an accurate diagnosis . In addition to a detailed medical history, skin biopsy is mandatory because the skin eruptions are not always unequivocal . Discontinuation of the causing agent is crucial, and treatment in specialized intensive care units or burn units is supportive . Currently there is no specific treatment for toxic epidermal necrolysis . Advantages from corticosteroids, plasmapheresis, intravenous immunoglobulin, cyclophosphamide, cyclosporin, and N-acetylcysteine still remain to be established by controlled trials, or have failed to prove a benefit (thalidomide) . The patient presented here demonstrates the difficulties in diagnosing toxic epidermal necrolysis in a critically ill patient . A short overview of the pathogenesis and the management of toxic epidermal necrolysis is provided.

AJNR Am J Neuroradiol, 2002 Aug, 23(7), 1117 - 20
Vertebroplasty in the mid- and upper thoracic spine; Kallmes DF et al.; BACKGROUND AND PURPOSE: Vertebroplasty performed in the mid- and upper thoracic spine presents technical challenges that differ from those in the lower thoracic and lumbar region . We herein report results of percutaneous vertebroplasty for treatment of painful, osteoporotic compression fractures in the mid- and upper thoracic spine . METHODS: Retrospective chart review identified vertebroplasty treatments performed for painful osteoporotic compression fractures at T4-T8 . The following were catalogued: percentage of vertebral body compression, needle size, surgical approach, clinical outcome, volume of cement injected, and complications . RESULTS: Sixty-three vertebral bodies were treated in 41 patients . Mean percentage of compression was 44% . Bipediculate injections were used in 12 (19%) of 63 treatment levels, and unipediculate injections were used in 51 (81%) of treatments (75 injections performed) . Eleven-gauge needles were used for 55 (73%) of the 75 injections, and 13-gauge needles were used for 20 (27%) . Clinical follow-up was available for 76% of the patients . Mean pre- and postoperative pain intensity was 9.7 +/- 1.0 and 1.7 +/- 1.9, respectively (P <.0001) . Mean pre- and postoperative medication scores were 3.4 +/- 0.7 and 1.7 +/- 1.7, respectively (P =.075) . Fracture involving the pedicle used for needle access was noted in one (1.3%) of 75 injections; this pedicle had been traversed using a 13-gauge needle . Staphylococcus epidermidis infection occurred in one case . No cases of pneumothorax were noted . CONCLUSION: Transpedicular vertebroplasty is readily and safely performed using 11-gauge needles in the mid- and upper thoracic regions, yielding excellent pain relief and low complication rates.

FEMS Microbiol Lett, 2002 Aug 6, 213(2), 193 - 7
Several regions of the repeat domain of the Staphylococcus caprae autolysin, AtlC, are involved in fibronectin binding; Allignet J et al.; The autolysin AtlC is the only known fibronectin-binding protein in Staphylococcus caprae strain 96007 . The fibronectin-binding domain of AtlC consists of three repeats (AtlCR(1)R(2)R(3)), which are located between the two enzymatic domains . The AtlCR(1)R(2)R(3) domain and the AtlCR(1)R(2) and AtlCR(3) subdomains were expressed separately as His(6)-tagged proteins . In Western affinity blots, only AtlCR(1)R(2)R(3) and AtlCR(3) but not AtlCR(1)R(2) appeared to recognise fibronectin; however, in ELISA and Biacore experiments, all three bound fibronectin . The interaction between AtlCR(1)R(2)R(3) and fibronectin is multivalent and involves high- and low-affinity sites that are present in a 2:1 ratio . These distinct classes of binding sites may be situated on either or on both ligands.

Mult Scler, 2002 Aug, 8(4), 307 - 9
Immunomodulatory effects of glatiramer acetate on superantigen- and mitogen-induced T-cell stimulation in vitro; Schmidt S et al.; Glatiramer acetate (GA) interferes with antigen recognition and modulates cytokine secretion of T cells in an antigen-specific manner . Here we analysed the capacity of GA to modulate proliferative responses and cytokine secretion of peripheral blood mononuclear cells (PBMCs) in response to antigen-independent stimuli, i.e., phytohaemagglutinin (PHA) and staphylococcal enterotoxin B (SEB) stimulation in five healthy volunteers . A significant reduction of proliferative responses, as well as interferon-gamma (IFNalpha) and tumour necrosis factor-alpha (TNFalpha) secretion, was observed at concentrations of 200 microg/ml suggesting that GA may also exert immunomodulatory effects on mitogen- and superantigen-induced T-cell stimulation in vitro . However, since systemic GA concentrations of this magnitude are highly unlikely to occur in vivo the immunomodulatory effects observed here are not likely to contribute to the therapeutic mechanisms of action under physiological conditions.

J Immunol, 2002 Aug 15, 169(4), 1774 - 83
Expression and function of transgenic HLA-DQ molecules and lymphocyte development in mice lacking invariant chain; Rajagopalan G et al.; Invariant chain (Ii) is a non-MHC-encoded molecule, which plays an accessory role in the proper assembly/expression of functional MHC class II molecules and there by plays an important role in Ag processing/presentation . The phenotype of mice lacking Ii depends on the allotype of the MHC class II molecule . In some mice strains, Ii deficiency results in reduction in expression of class II molecules accompanied by defective CD4(+) T cell development . Responses to conventional Ags/superantigens are also compromised . In this study, we describe for the first time the functionality of human class II molecules, HLA-DQ6 and HLA-DQ8, in transgenic mice lacking Ii . HLA transgenic Ii(-/-) mice expressed very low levels of surface DQ6 and DQ8 accompanied by severe reduction in CD4(+) T cells both in the thymus and periphery . In vitro proliferation and cytokine production to an exogenous superantigen, staphylococcal enterotoxin B (SEB) was diminished in HLA-transgenic Ii(-/-) mice . However, SEB-induced in vivo expansion of CD8(+) T cells expressing TCR Vbeta8 family in DQ8.Ii(-/-) mice was comparable with that of DQ8.Ii(+/+) mice . Systemic IFN-gamma production following in vivo challenge with SEB was reduced in DQ8.Ii(-/-) mice and were also protected from SEB-induced toxic shock . Although the T cell response to a known peptide Ag was diminished in DQ8.Ii(-/-) mice, DQ8.Ii(-/-) APCs were capable of presenting that peptide to primed T cells from wild-type DQ8 mice as well as to a specific T cell hybridoma . Differentiation of mature B cells was also affected to a certain extent in DQ8.Ii(-/-) mice.

Acta Paediatr, 2002, 91(6), 670 - 3
Intravenous rifampicin in neonates with persistent staphylococcal bacteraemia; Shama A et al.; Addition of intravenous rifampin is reported to be useful in prompt clearance of persistent coagulase negative staphylococcal (CONS) bacteraemia in high-risk neonates . Four neonates (mean birthweight 823 g, mean gestation 25 wk) with persistent CONS bacteraemia for > 7-10 d (mean 11) were treated with i.v . rifampicin (10 mg/kg/12 h x 10 d) while continuing vancomycin (15 mg/kg/24 h) . Their age at time of infection ranged from 2 to 11 d . The mean (range) vancomycin peak and trough concentrations were 29 (25-35) and 6 (4-10) microg/ml, respectively . The blood isolates were Staphylococcus epidermidis, S . hominis, and S . haemolyticus . Addition of rifampicin was associated with prompt clearance of bacteraemia within 48 h (n = 3) and 5 d (n - 1) . Rifampicin-related adverse effects such as abnormal liver function tests and thrombocytopenia did not occur . CONCLUSION: Addition of i.v . rifampicin to vancomycin may optimize the outcome of persistent CONS bacteraemia and the risk of bacterial resistance related to prolonged exposure to vancomycin.

Biomedica, 2002 Jun, 22(2), 110 - 5
{Chronic bronchial dilatations in different colonies of laboratory rats}; Miguel JC et al.; Bronchiectasis occurred naturally in 12-month-old spontaneously diabetic eSS male rats . The lungs of 3 and 6-month-old eSS rats were compared in eumetabolic eSS rats from three inbred lines consisting of inbred spontaneously diabetic eSS derived from IIM strain; these were compared with eumetabolic, outbred Wistar rats, paired by sex and age . Acrylic casts of bronchial tree were obtained after injection of a plastic substance . The casts were pruned to focus on the first four bronchial branchings . Diameter and volume of the conductive bronchial tree were determined using a binocular magnifier . Histological sections were obtained . All lines showed multiple bronchiectasis, mostly fusiform, bronchial dilatation and inflammatory response with lymphocytic infiltrates . These symptoms were much more severe in 180-day-old eSS males . Bacteria were isolated from the lungs in 70% of cases (n = 32), except in eSS rats . Pseudomonas spp . (38%) and Gram-positive cocci as coagulase-negative Staphylococcus spp . (20%) were detected . Neither pathogenic bacteria nor saprophyte fungi were found . Although all lines were affected, diabetes in eSS appears to be an aggravating factor.

Lijec Vjesn, 2002 May, 124(5), 134 - 6
{Detection of Staphylococcus lugdunensis in clinical specimens and their sensitivity to antimicrobial agents}; Vukadinovic MV et al.; Staphylococcus lugdunensis is a newly recognised coagulase-negative staphylococcus species, pathogenic for humans . Following patients' records over a 25 month period, we revealed 25 cultures positive for S . lugdunensis . Most of the samples were from infected wounds or abscesses in patients suffering from angiopathies of lower limbs, late complications of diabetes . SL was also isolated from a chronic breast abscess and ascites in another patient with liver cirrhosis . In five cases we understood the isolate as normal skin flora or colonising organism mostly in mixed culture . Resistance to penicillin was found in 6/25 isolates, and resistance to azithromycin, chloramphenicol and/or gentamicin was less common.

Ear Nose Throat J, 2002 Jul, 81(7), 458 - 61
Microbiology of the ethmoid sinus following endoscopic sinus surgery; Bhattacharyya N et al.; We prospectively studied the native microbiology of the ethmoid sinus following endoscopic sinus surgery in 113 patients (mean age: 41.3 yr) . After each patient had regained complete mucosalization (minimum follow-up: 6 wk), we obtained a bacterial swab of the ethmoid sinus and submitted it for culture and sensitivity analysis . We then compiled data on sensitivity patterns and the number and type of bacteria isolated . Of the 113 patients, 67 (59.3%) had positive cultures, 26 (23.0%) had sterile cultures, and 20 (17.7%) had normal flora . Multiple bacteria were isolated in 31 of the 113 patients (27.4%) . The most common isolates were gram-positive cocci, and the most common bacteria were staphylococcal species . A significant degree (42.9%) of beta-lactamase resistance was exhibited . We conclude that the ethmoid sinus is not microbiologically sterile following endoscopic sinus surgery.

J Clin Microbiol, 2002 Aug, 40(8), 2936 - 41
Mediastinitis after cardiac surgery: improvement of bacteriological diagnosis by use of multiple tissue samples and strain typing; Tammelin A et al.; The diagnosis of postsurgical mediastinitis (PSM) among patients with sternal wound complication (SWC) after cardiac surgery is sometimes difficult, as fever, elevated C-reactive protein levels, and chest pain can be caused by a general inflammatory reaction to the operative trauma and/or sternal dehiscence without infection . The definitions of PSM usually used emphasize clinical signs and symptoms easily observed by the surgeon . The aim of the study was to investigate whether the use of standardized multiple tissue sampling, optimal culturing methods, and strain typing, together with a microbiological criterion for infection, could identify more infected patients than clinical assessment alone . Patients reexplored due to SWC after cardiac artery bypass grafting (CABG) or heart valve replacement (HVR) with or without CABG performed at the Department for Cardio-Thoracic Surgery at the Uppsala University Hospital between 10 March 1998 and 9 September 2000 were investigated prospectively . Tissue samples were taken from the sternum or adjacent mediastinal tissue, preferably before the administration of antibiotics . Culturing was performed both directly (on agar plates) and using enrichment broth . Species identification was performed by standard methods, and strain typing was performed by pulsed-field gel electrophoresis . A total of 41 cases with at least five tissue samples each were included in the study group . Of these patients, 32 were infected according to the microbiological criterion (i.e., the same strain was found in >/=50% of the samples) . Staphylococcus epidermidis was the primary pathogen in 38% of the cases (12/32), S . aureus was the primary pathogen in 31% (10/32), P . acnes was the primary pathogen in 25% (8/32), and S . simulans and S . haemolyticus were the primary pathogens in 3% (1/32) each . All cases of S . aureus infection and 86% (12/14) of coagulase-negative staphylococcus (CoNS) infections were identified from primary cultures . All cases fulfilling the microbiological criterion for S . aureus infection were clinically diagnosed as cases of infection, but among the 14 cases fulfilling the criterion for microbiological diagnosis of CoNS infection, only 10 appeared to qualify clinically as cases of infection . Among the patients with sternal dehiscence in whom a microbiological diagnosis was established, 67% (12/18) had a CoNS infection, compared to 14% (2/14) of those without sternal dehiscence . The difference was statistically significant . PSM caused by S . aureus is readily identified by the surgeon, whereas 30% of cases with CoNS infections may be misinterpreted as noninfected . Multiple sampling before administration of antibiotics, primary culturing on agar plates, species identification, strain typing, and susceptibility testing should be used to ensure a fast and microbiologically correct diagnosis which identifies the primary pathogen and infected patients among those with minor infective symptoms . The role of P . acnes as a possible cause of PSM needs further investigation . PSM caused by CoNS is significantly related to sternal dehiscence.

Clin Lab, 2002, 48(7-8), 395 - 400
Detection of staphylococcal enterotoxin B (SEB) by enzyme-linked immunosorbent assay and by a rapid hand-held assay; Schotte U et al.; Staphylococcal enterotoxins are a frequent cause of food poisoning . Immunologically-based assays for the detection of staphylococcal enterotoxins are commercially available, but require at minimum of 3 hours . We used staphylococcal enterotoxin B to compare two commercially available assays with a newly developed rapid immunochromatographic-based hand-held assay . The detection limit of the commercially available assays accounted for 500 pg and 100 pg enterotoxin B/ml, respectively, whereas 50 pg enterotoxin B /ml were detected within 15 min using the hand-held assay . Enterotoxin B-spiked custard served as a model to detect staphylococcal enterotoxin in food . Depending on the extraction method applied, the detection limit was in the range of 500 pg to 2500 pg/g custard for the commercially available assays and 62.5 pg/g for the hand-held assay . We conclude that the hand-held assay is widely applicable because it is sensitive, specific with regard to the tested enterotoxins, and the results can be read with the naked eye.

Chem Biol, 2002 Jul, 9(7), 829 - 38
Stochastic sensing of nanomolar inositol 1,4,5-trisphosphate with an engineered pore; Cheley S et al.; The introduction of a ring of arginine residues near the constriction in the transmembrane beta barrel of the staphylococcal alpha-hemolysin heptamer yielded a pore that could be almost completely blocked by phosphate anions at pH 7.5 . Block did not occur with other oxyanions, including nitrate, sulfate, perchlorate, and citrate . Based on this finding, additional pores were engineered with high affinities for important cell signaling molecules, such as the Ca(2+)-mobilizing second messenger inositol 1,4,5-trisphosphate (IP(3)), that contain phosphate groups . One of these engineered pores, P(RR-2), provides a ring of fourteen arginines that project into the lumen of the transmembrane barrel . Remarkably, P(RR-2) bound IP(3) with low nanomolar affinity while failing to bind another second messenger, adenosine 3', 5'-cyclic monophosphate (cAMP) . The engineered alpha-hemolysin pores may be useful as components of stochastic sensors for cell signaling molecules.

Chem Senses, 2002 Jul, 27(6), 521 - 7
Cell death of olfactory receptor neurons in a rat with nasosinusitis infected artificially with Staphylococcus; Ge Y et al.; Nasosinusitis is a common cause of acquired hyposmia or anosmia . To study the apoptotic death of olfactory receptor neurons in nasosinusitis, we made an inflammation model in rat infected with STAPHYLOCOCCUS: The histochemical changes in olfactory epithelium were examined using antibodies against protein gene product 9.5 (PGP 9.5), single-strand DNA (ssDNA), Bcl-2 and Bax that might be involved in the apoptosis of olfactory receptor neurons . The thickness of olfactory epithelium and the number of ssDNA-labeled cells were evaluated in each post-treatment group and the results were analyzed by two-way analysis of variance (ANOVA) and post hoc tests . Hematoxylin-eosin staining showed that a severe inflammatory reaction had occurred on the infected side of the nasal cavity and sinus, but not on the non-infected side . However, apoptosis of olfactory receptor neurons occurred on both sides; the apoptosis on the non-infected side started later and behaved like a shadow curve similarly to the infected side . Repeated measures ANOVA showed significant differences of both the thickness of olfactory epithelium (P < 0.0001) and the number of ssDNA-labeled cells (P = 0.0339) in the epithelium between the infected side and non-infected side comparing treatment, time and their interactions . Bcl-2 and Bax were detected only on the infected side in the early stages . Thus, nasosinusitis induced the apoptosis of olfactory receptor neurons . However, the apoptosis occurred not only on the infected side, but also on the non-infected side with no significant inflammation . The Bcl-2/Bax family seems to play an important role in the apoptosis induced by infection, but not in the apoptosis on the non-infected side . The results suggest that mechanisms of apoptosis of olfactory receptor neurons on the infected side may differ from those on the non-infected side.

Zh Mikrobiol Epidemiol Immunobiol, 2002 May-Jun, (3), 48 - 51
{Staphylococcal colonization of the oral cavity of children following an acute form of rubella infection}; Kolenchukova OA et al.; The children were being observed during the course of acute rubella infection and postinfectious period . Quantitative dynamic changes in the normal microflora composition were noted weekly during the first month of the disease as well as in the beginning of the second and the third months . The amount of microorganisms registered 3 weeks and in the beginning of the second month after the onset of rubella infection differed from that in the controls . The identification of the Staphylococcus genus microorganisms has shown an increased number of carriers during the first week, as well as on the third week from the onset of the disease . During these periods a secondary immunodeficiency is supposed to develop in children resulting in exacerbation of bacterial infections.

Proc Natl Acad Sci U S A, 2002 Aug 6, 99(16), 10394 - 8 Epub 2002 Jul 24.
An atomically detailed study of the folding pathways of protein A with the stochastic difference equation; Ghosh A et al.; An algorithm is applied here to compute folding pathways of staphylococcal protein A, fragment B . Emphasis is on studies of the complete process, starting from an ensemble of fully denatured conformations and ending at the folded state . The stochastic difference equation algorithm is based on optimization of an action that makes it possible to use a large integration step . Motions with typical displacements that change rapidly on the size scale of the step are filtered out, providing numerically stable and approximate solutions . The present approach is unique in maintaining an atomically detailed picture while providing a systematic, controlled approximation to the classical equations of motion . Analysis of 130 trajectories suggests the following folding mechanism for protein A: At an early precollapse phase of the process, a few native hydrogen bonds form near the C terminus of the protein . The hydrogen bonds are formed mostly within the third helix . The next step is chain collapse that occurs in parallel to additional growth of secondary structure seeds . Therefore, the present study does not support a pure hydrophobic collapse, or substantial early formation of secondary structure . At the last step, native tertiary contacts are formed at the same time as the completion of the secondary structure elements . To a large extent, the process is parallel and not sequential . The early formation of the third helix of protein A, fragment B (in the calculation), is consistent with experimental data.

Am J Ophthalmol, 2002 Aug, 134(2), 274 - 5
Vitreous hemorrhage as the initial presentation of postoperative endophthalmitis; Tewari A et al.; PURPOSE: To describe a patient with postoperative endophthalmitis whose only abnormal finding at presentation was a vitreous hemorrhage . DESIGN: Interventional case report . METHODS: A 68-year-old diabetic woman underwent cataract extraction with intraocular lens implantation in the left eye . Three days after surgery, she had painless loss of vision, minimal anterior chamber inflammation, and dense vitreous hemorrhage in the left eye . RESULTS: On the fourth postoperative day, significant anterior chamber inflammation developed with fibrin and a hypopyon . During vitrectomy with intravitreal antibiotic injection, an area of retinitis surrounding an eroded retinal blood vessel was found . Cultures of undiluted vitreous fluid grew coagulase-negative Staphylococcus organisms . The endophthalmitis resolved and 20 months later, her best-corrected visual acuity had improved to 20/40 . CONCLUSION: Postoperative endophthalmitis may present as a vitreous hemorrhage, secondary to retinitis and erosion of a retinal blood vessel.

Graefes Arch Clin Exp Ophthalmol, 2002 Jul, 240(7), 557 - 64 Epub 2002 Jun 20.
Intravenous imipenem prophylaxis in experimental endophthalmitis; Mino de Kaspar H et al.; PURPOSE: To determine the effect of prophylactic intravenous imipenem on experimental Staphylococcus aureusendophthalmitis . METHODS: The right eyes of 36 New Zealand albino rabbits received an intraocular injection of 100, 1,000, or 10,000 colony-forming units (CFU) of S . aureus . Each of these three groups was subdivided into a control group (no antibiotic prophylaxis) and a prophylactic group, which received 37.5 mg/kg imipenem 8 h prior to inoculation of bacteria, 30 min prior to inoculation of bacteria, and 8 h after inoculation of bacteria . Eyes were observed daily by slit-lamp examination and funduscopy . The eyes were enucleated after 5 and 9 days for the control and prophylactic groups, respectively . Vitreous samples were cultured for bacteria, and the eyes were prepared for histologic evaluation . RESULTS: All eyes that received imipenem demonstrated significantly less inflammation (two-way analysis of variance; P<0.001) on clinical examinations than did control eyes . The prophylactic groups that had received 100 CFU and 1,000 CFU of bacteria also had significantly fewer positive vitreous cultures than the corresponding controls (chi(2) test; P<0.001 and P<0.01, respectively) . Histologically, the prophylactic groups injected with 100 CFU and 1,000 CFU of bacteria had significantly less inflammation than the control groups (Student's t-test; P<0.00001 and P<0.0001, respectively) . There was no significant difference in the number of positive vitreous cultures or histologically between the control and prophylactic groups that received 10,000 CFU of bacteria . CONCLUSIONS: Prophylactic intravenous imipenem can prevent or significantly reduce the severity of experimental endophthalmitis.

Am J Surg, 2002 Jul, 184(1), 37 - 40
Mild hypothermia does not increase bacterial proliferation on implanted vascular grafts; Alfonsi P et al.; BACKGROUND: Mild hypothermia may offer protection against spinal cord ischemia during aortic surgery . However, hypothermia also promotes postoperative infection via two mechanisms: peripheral vasoconstriction and impairment of various immune functions . If mild hypothermia aggravates graft infections, immune function impairment would presumably be the most important factor because thermoregulatory vasoconstriction does not appreciably reduce aortic blood flow . We therefore tested the hypothesis that resistance to vascular graft infection is not reduced by mild perioperative hypothermia in dogs . METHODS: After colonization with a solution of Staphylococcus epidermidis, prostheses were used to replace the infrarenal aorta in 20 dogs . During surgery, the dogs were randomly assigned to maintain of normothermia or passive cooling . Seven days later, grafts were recovered for bacteriologic study . RESULTS: Colony counts for the grafts removed from the normothermic and hypothermic dogs did not differ significantly . CONCLUSIONS: Mild perioperative hypothermia does not increase proliferation of S epidermidis on aortic vascular grafts.

Immunity, 2002 Jun, 16(6), 759 - 67
Activated T cell death in vivo mediated by proapoptotic bcl-2 family member bim; Hildeman DA et al.; At the end of the T cell response, the majority of the activated T cells die . We activated Vbeta8(+) T cells with staphylococcal enterotoxin B (SEB) in vivo and monitored the expansion and deletion of Vbeta8(+) T cells . We found that, in response to SEB, activated T cells died in vivo in the absence of Fas or TNF-R signaling but not when they overexpressed human Bcl-2 . We also found that Vbeta8(+) T cells from Bim-deficient mice are resistant to SEB-induced deletion . While Bim levels did not change, endogenous Bcl-2 levels within Vbeta8(+) T cells decrease following SEB injection . Thus, the death of superantigen-stimulated T cells in vivo is mediated by Bim and may be modulated by a decrease in Bcl-2.

Australas J Dermatol, 2002 Aug, 43(3), 199 - 201
Staphylococcal scalded skin syndrome complicating acute generalized pustular psoriasis; Sharkey MP et al.; A 60-year-old woman with psoriasis vulgaris treated with oral cyclosporin and acitretin developed an acute generalized pustular eruption with erythema and associated fever consistent with acute generalized pustular psoriasis . She was admitted to hospital and, despite intravenous fluid replacement, developed acute renal failure . In addition, she developed staphylococcal septicaemia . After transfer to the intensive care unit because of deteriorating renal function, a sudden onset of widespread flaccid blistering (Nikolsky sign positive) and superficial erosions was noted . Histology of a biopsied blister revealed subcorneal splitting of the epidermis consistent with staphylococcal scalded skin syndrome . The patient was treated with intravenous dicloxacillin and the blistering gradually improved over 10 days.

Infect Immun, 2002 Aug, 70(8), 4148 - 57
Mycobacterium bovis BCG-infected mice are more susceptible to staphylococcal enterotoxin B-mediated toxic shock than uninfected mice despite reduced in vitro splenocyte responses to superantigens; Pedras-Vasconcelos JA et al.; Type 1 T-cell responses against intracellular pathogens play a crucial role in mediating protection . We examined whether the induction of a strong type 1 T-cell response during a chronic bacterial infection influences responses to superantigens capable of inducing acute shock . Intravenous infection of mice with Mycobacterium bovis BCG appeared to induce a progressive anergy towards staphylococcal enterotoxin B (SEB) and towards antigen preparation of BCG (BCG-Ag) itself, based on diminished gamma interferon (IFN-gamma) production by SEB- and BCG-Ag-stimulated splenocytes from infected mice . In contrast to these in vitro results, injection of SEB into BCG-infected mice led to a dramatic increase in the serum IFN-gamma levels and the death of infected but not of control mice . In vitro hyporesponsiveness towards SEB and BCG-Ag occurred only with unfractionated splenocyte cultures, as purified T cells from infected mice produced higher levels of IFN-gamma . Hyporesponsiveness towards SEB and BCG-Ag in unfractionated splenocyte cultures was not due to suppressive antigen-presenting cells (APCs), as APCs from infected mice stimulated higher levels of IFN-gamma from purified T cells . The diminished IFN-gamma levels observed with bulk splenocytes appear to be due to changes in the T-cell-to-APC ratio that result in a decreased proportion of T cells, coupled to reduced proliferative responses and an increased susceptibility of effector T cells to activation-induced cell death in vitro . Our results indicate that the reported phenomena of T-cell anergy during mycobacterial infection may be an in vitro consequence of the development of a strong type 1 response in vivo.

Proteins, 2002 Aug 1, 48(2), 283 - 92
The role of hydrophobic microenvironments in modulating pKa shifts in proteins; Mehler EL et al.; The screened Coulomb potential (SCP) method, combined with a quantitative description of the microenvironments around titratable groups, based on the Hydrophobic Fragmental Constants developed by Rekker, has been applied to calculate the pK(a) values of groups embedded in extremely hydrophobic microenvironments in proteins . This type of microenvironment is not common; but constitutes a small class, where the protein's architecture has evolved to lend special properties to the embedded residue . They are of significant interest because they are frequently important in catalysis and in proton and electron transfer reactions . In the SCP treatment these special cases are treated locally and therefore do not affect the accuracy of the pK(a) values calculated for other residues in less hydrophobic environments . Here the calibration of the algorithm is extended with the help of earlier results from lysozyme and of three mutants of staphylococcal nuclease (SNase) that were specially designed to measure the energetics of ionization of titratable groups buried in extremely hydrophobic microenvironments . The calibrated algorithm was subsequently applied to a fourth mutant of SNase and then to a very large dimeric amine oxidase of 1284 residues, where 334 are titratable . The observed pK(a) shifts of the buried residues are large (up to 4.7 pK units), and all cases are well reproduced by the calculations with a root mean square error of 0.22 . These results support the hypothesis that protein electrostatics can only be described correctly and self-consistently if the inherent heterogeneity of these systems is properly accounted for .

Proteins, 2002 Aug 15, 48(3), 454 - 62
Anti-idiotypic protein domains selected from protein A-based affibody libraries; Eklund M et al.; Three pairs of small protein domains showing binding behavior in analogy with anti-idiotypic antibodies have been selected using phage display technology . From an affibody protein library constructed by combinatorial variegation of the Fc binding surface of the 58 residue staphylococcal protein A (SPA)-derived domain Z, affibody variants have been selected to the parental SPA scaffold and to two earlier identified SPA-derived affibodies . One selected affibody (Z(SPA-1)) was shown to recognize each of the five domains of wild-type SPA with dissociation constants (K(D)) in the micromolar range . The binding of the Z(SPA-1) affibody to its parental structure was shown to involve the Fc binding site of SPA, while the Fab-binding site was not involved . Similarly, affibodies showing anti-idiotypic binding characteristics were also obtained when affibodies previously selected for binding to Taq DNA polymerase and human IgA, respectively, were used as targets for selections . The potential applications for these types of affinity pairs were exemplified by one-step protein recovery using affinity chromatography employing the specific interactions between the respective protein pair members . These experiments included the purification of the Z(SPA-1) affibody from a total Escherichia coli cell lysate using protein A-Sepharose, suggesting that this protein A/antiprotein A affinity pair could provide a basis for novel affinity gene fusion systems . The use of this type of small, robust, and easily expressed anti-idiotypic affibody pair for affinity technology applications, including self-assembled protein networks, is discussed .

Clin Exp Immunol, 2002 Jun, 128(3), 453 - 9
Activation of human T cells with NK cell markers by staphylococcal enterotoxin A via IL-12 but not via IL-18; Ami K et al.; We have reported recently that mouse liver NK cells and NK1 x 1+ T cells were activated by bacterial superantigens via the IL-12 production from Kupffer cells . In the present study, we examined the effect of staphyloccoccal enterotoxin A (SEA) on human T cells with NK cell markers, CD56 or CD57 (NK-type T cells) . After stimulating peripheral blood mononuclear cells (PBMC) with SEA, PBMC produced a large amount of IFN- and acquired a potent antitumour cytotoxicity . The in vitro depletion of either CD56+ TCR NK cells, CD56+ T cells or 57+ T cells from PBMC significantly inhibited the IFN- production from PBMC . When purified NK-type T cells, NK cells and regular T cells were cultured with monocytes and SEA they all produced IFN-, while the IFN- amounts produced by both NK-type T cells were greater than those produced by NK cells . NK cells as well as CD56+ T cells showed cytotoxicity against NK-sensitive K562 cells, whereas both NK-type T cells showed a more potent cytotoxicity against NK-resistant Raji cells than did NK cells . The IFN- production from each population as well as from whole PBMC was greatly inhibited by anti-IL-12 antibody but not by anti-IL-18 antibody . The antitumour cytotoxicity of whole PBMC was also significantly inhibited by anti-IL-12 antibody while the SEA-induced proliferation of PBMC was not affected by anti-IL-12 antibody . Furthermore, SEA-activated NK-type T cells as well as NK cells showed cytotoxicities against vascular endothelial cells . Our findings suggest that human NK-type T cells are thus involved in bacterial superantigen-induced immune response.

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