Acta Microbiol Immunol Hung, 2002, 49(2-3), 363 - 9 Pathological and clinical aspects of the diseases caused by Malassezia species; Dorogi J; From veterinary point of view Malassezia pachydermatis has the greatest significance . It has been standing in the focus of interest since the early 1990s, mostly because of the frequency of otitis externa and dermatitis caused by this yeast in dogs . This is the only lipid-independent species in the genus Malassezia . It can be found in very large proportion on the skin of healthy animals, but can be isolated in much greater number from diseased dogs . It often causes illness together with other pathogens (e.g . Staphylococcus intermedius) . Some breeds are predisposed . In addition to the treatment of the accidental concurrent diseases, therapy consists of systemic and/or topical antimicrobial treatment . Ketoconazole is used most frequently . Malassezia pachydermatis plays also a role in the skin disorders of other carnivores . It has little zoonotic potential, it can be dangerous to immunocompromised humans . The other Malassezia species have little veterinary importance, although M . sympodialis and M . globosa were isolated from asymptomatic animals (mostly cats) and from mixed infections. Przegl Lek, 2002, 59 Suppl 1, 50 - 3 {Quantitative changes of CD3+, CD4+, CD8+, HLA-DR+, T lymphocytes and CD25+ cells in eutrophic full-term neonates with staphylococcal septicemia}; Behrendt J et al.; OBJECTIVE: Influence of staphylococcal septicemia on changes of share and changes of amount of CD3+ lymphocytes and their subpopulation as well as CD25+ cells in eutrophic full-term neonates . MATERIALS AND METHODS: In 52 full-term neonates, with birthweight ranged from 2900 to 4500 g, including 30 infants with staphylococcal septicemia caused by Staphylococcus epidermidis (19 cases), S . sciuri (2 cases) S . varneri, S . hominis, S . haemolyticus and S . aureus (6 cases) and 22 healthy neonates (control group) the subpopulation of CD3+, CD4+, CD8+, HLA-DR+ lymphocytes in venous blood was estimated using flow cytometer FACScan with monoclonal antibodies of Becton Dickinson . RESULTS: Average percentage of CD3+ (69.2 +/- 5.9%), CD4+ (48.9 +/- 10.7%) and HLA-DR+ (1.8 +/- 0.9%) lymphocytes and average CD4+/CD8+ ratio (2.97 +/- 1.33) in neonates with septicemia did not significantly differ from average values of these parameters in healthy neonates that were: CD3+ (69.1 +/- 9.0%), CD4+ (47.4 +/- 10.6%), HLA-DR+ (1.5 +/- 0.5%) and CD4+/CD8+ (2.29 +/- 1.29) . Likewise the average numbers of these lymphocytes were close to the values found in the control group . Whereas the average subpopulation of CD8+ (19.2 +/- 4.6%) lymphocytes in ill neonates was significantly lower (p = 0.007) than in healthy ones (23.6 +/- 6.6%), and the average number of CD25+ cells was essentially (p = 0.01) higher in septicemia neonates (8.5 +/- 2.5) compared to the control group (6.6 +/- 1.3%) . Average numbers of these cells in ill neonates did not substantially differ from their average values in control group . CONCLUSIONS: 1 . Staphylococcal septicemia significantly decreases the values of subpopulation of CD8+ lymphocytes and increases the number of CD25+ cells in eutrophic full-term neonates . 2 . Estimation of lymphocytes, and their subpopulation and CD25+ cells in neonates with staphylococcal septicemia may be useful in assessment of immunological changes in severe infections. Surg Today, 2002, 32(6), 566 - 7 Ruptured staphylococcal splenic abscess resulting in peritonitis: report of a case; Balasubramanian SP et al.; A 50-year-old man presented with features of peritonitis of 2 days duration . The signs were more marked in the left upper abdomen . Investigations followed by a laparotomy showed a ruptured splenic abscess, the cause of which was not apparent . We herein present the case report of this unusual cause of peritonitis along with a relevant review of the literature. J Bone Joint Surg Am, 2002 Jul, 84-A(7), 1138 - 41 Total knee arthroplasty in hemophilic arthropathy; Norian JM et al.; BACKGROUND: Arthropathy of the knee frequently develops in patients with hemophilia, who may require a total knee arthroplasty at a young age . Hemophilic patients, who require regular intravenous replacement of coagulation factor, have a higher prevalence of human immunodeficiency virus (HIV) infection, which can compromise the outcome of the arthroplasty . The purpose of this study was to evaluate prosthetic survival following total knee arthroplasty and identify factors associated with failures of the arthroplasties in hemophilic patients . METHODS: The results of fifty-three total knee arthroplasties performed in thirty-eight patients (twenty-nine of whom were seropositive for HIV) to treat hemophilic arthropathy between 1976 and 1998 were retrospectively reviewed . Inpatient and outpatient medical records were studied to determine the HIV status, CD4 lymphocyte count, type of prosthesis, duration of prosthetic survival, cause of failure, and cause of death . If an arthroplasty failed, the outcome of the treatment of the failed arthroplasty was also determined . RESULTS: The rate of survival of the prostheses was 90% after five years . Eleven total knee arthroplasties failed . The most common cause of failure was infection (seven knees), which developed at an average of sixty months (range, three to 138 months) after the arthroplasty . There was no significant difference in the CD4 lymphocyte counts between the patients in whom infection developed and those in whom it did not . The HIV status also did not appear to be related to the development of infection . Thirteen patients died, and the most common cause of death was complications associated with acquired immunodeficiency syndrome (AIDS) . CONCLUSIONS: Total knee arthroplasty performed to treat hemophilic arthropathy has a high risk of failure as a result of infection . Most infections developed late and were frequently caused by Staphylococcus epidermidis, suggesting that a likely cause of failure due to infection was hematogenous spread during administration of coagulation factor . It may be difficult to salvage a prosthesis complicated by infection . However, the life expectancy of hemophilic patients is lower than that of the general population of patients treated with total knee arthroplasty, and the improvement in the quality of life after total knee arthroplasty for hemophilic arthropathy may outweigh the risk of failure. Vox Sang, 2002 Jul, 83(1), 13 - 6 Diversion of first blood volume results in a reduction of bacterial contamination for whole-blood collections; de Korte D et al.; BACKGROUND AND OBJECTIVES: In a previous study we established a reliable setpoint for the prevalence of bacteria in whole blood . In the present study we investigated the possible preventive effect, of diversion of the first 10 ml of a blood donation, on the bacterial contamination rate . MATERIALS AND METHODS: To divert the first 10 ml of a whole-blood donation, we used a special five-bag system equipped with a Composampler device . After venepuncture, the first 10 ml of a donation was sampled into a vacutainer tube . This was followed by the collection of the whole-blood unit . The extra bag allowed direct sampling of the final donation in a closed system for BacT/Alert . Whole-blood samples were taken after storage (2-14 h at 20 degrees C) and subsequent mixing . BacT/Alert culture bottles were incubated until positive, or for 7 days if negative . Confirmation and identification of positive cultures was performed according to internationally recognized standard reference methods . RESULTS: The prevalence of bacteria in whole blood, as determined by using standard collection techniques, was 0.35% (95% confidence interval 0.27-0.44%, n = 18 257) . After diversion of the first 10 ml this value was significantly lower: 0.21% (P < 0.05, 95% confidence interval 0.12-0.35%, n = 7087) . Most strikingly, a reduction in the frequency of staphylococcal species was observed (P < 0.02, reduction from 0.14 to 0.03%) . CONCLUSIONS: Diversion of the first 10 ml of blood was shown to contribute significantly to a reduction in the prevalence of superficial skin bacteria in whole-blood units . In our opinion, blood collection systems should be adapted to use the first 10-30 ml of a whole-blood donation for testing purposes. Int Immunol, 2002 Jul, 14(7), 801 - 12 Staphylococcal enterotoxin B-induced activation and concomitant resistance to cell death in CD28-deficient HLA-DQ8 transgenic mice; Rajagopalan G et al.; HLA class II molecules present superantigens more efficiently than their murine counterpart . Therefore, transgenic mice expressing HLA-DQ8 with and without CD28 were used to address the role of CD28 in staphylococcal enterotoxin B (SEB)-driven immune responses . SEB-induced in vitro proliferation of naive DQ8.CD28(-/-) splenocytes was comparable to DQ8.CD28(+/+) cells, and was several fold higher than that of C57BL/10 and BALB/c splenocytes . SEB-activated, naive DQ8.CD28(-/-) cells in vitro produced significantly less IL-2, IL-4 and IL-10 than DQ8.CD28(+/+) cells, while IFN-gamma and IL-6 production was comparable . SEB-induced in vivo expansion of CD4(+) T cells and, to a greater extent, CD8(+) T cells was compromised in DQ8.CD28(-/-) mice, indicating that SEB-induced proliferation of CD8(+) T cells is more dependent on CD28 co-stimulation . Upon re-stimulation, SEB-primed CD28(+/+) T cells failed to proliferate but were capable of producing cytokines . Conversely, CD28(-/-) T cells were capable of proliferation, but not cytokine production . SEB-primed CD28-deficient cells produced significantly less nitric oxide when compared to CD28-sufficient cells following re-stimulation with SEB . CD28(+/+) and not CD28(-/-) mice were highly susceptible to SEB-induced lethal shock characterized by significantly elevated serum IFN-gamma . Thus, (i) efficient presentation of SEB by HLA-DQ8 circumvents co-stimulation through CD28, (ii) unique CD28-derived signals are mandatory for generation of certain effector functions, and (iii) absence of CD28-derived signals confers resistance to activation-induced cell death and protects mice from SEB-induced shock. J Infect Dis, 2002 Jul 1, 186(1), 15 - 22 Epub 2002 Jun 14. Dynamics of CD4 and CD8 T cell responses to cytomegalovirus in healthy human donors; Dunn HS et al.; To study the dynamics of cytomegalovirus (CMV) immunity in healthy immunocompetent hosts, interferon-gamma-producing CD4 and CD8 T cell responses in the presence or absence of CMV antigens were measured from 15 CMV-seropositive donors and 13 CMV-seronegative donors . Cytokine responses in the absence of antigen were significantly higher in CMV-seropositive donors . Also, a disproportionate number of CD69(+) cells isolated ex vivo from CMV-seropositive donors were specific for CMV, suggesting recent reactivation in vivo . To examine changes in cellular responses over time, 10 seropositive donors were tested over a 6-month period . About half of the donors showed significant variability over time, but staphylococcal enterotoxin B responses remained relatively constant . These findings suggest that CMV can present a considerable and recurrent burden to the human immune system . By understanding the normal dynamics of CMV responses over time, it may be possible to better identify aberrant responses to CMV in immunocompromised hosts. Chir Narzadow Ruchu Ortop Pol, 2002, 67(1), 61 - 7 {The treatment of acute hematogenous osteomyelitis in children and young adults}; Bielawski J et al.; On the basis of own experiences and literature from last three years authors presented the role of surgical treatment in children above two years and young people, suffering from acute haematogenous osteomyelitis . Conservative treatment should be limited to these cases, where we can see a quick recovery, reducing of the pain and decreasing of the level of CRP and ESR . The presence of subperiosteal abscessus, positive bacteriological culture from blood and other complications (i.e . staphylococcal pneumonia), are the absolute indications to decompression of the medullary cavity, and local application of antibiotics. Eur J Epidemiol, 2001, 17(8), 715 - 20 Neonatal late-onset bloodstream infection: attributable mortality, excess of length of stay and risk factors; Pessoa-Silva CL et al.; BACKGROUND: Bloodstream infection represents a major threat among neonates under intensive care with considerable impact on morbidity and mortality . This study evaluated extra stay, attributable mortality and the risk factors associated with late-onset bloodstream infection (LO-BSI) among neonates admitted to a neonatal intensive care unit during a 4-year period . METHODS: A retrospective matched cohort study was conducted . For each case, there was one control patient without LO-BSI matched for sex, birth weight, gestational age, duration of hospitalization prior to the date of LO-BSI in the respective cases, underlying illness and birth date . A novel test, sequential plan, was employed for attributable mortality analysis in addition to standard tests . Multiple logistic regression was employed for risk factor analysis . RESULTS: Fifty pairs of cases and controls were compared . LO-BSI prolonged hospital stay of 25.1 days in pairs where both subjects survived . Overall attributable mortality was 24% (95% CI: 9-39% p < 0.01) and specific attributable mortality due to Staphylococcus epidermidis was 26.7% (95% CI: 23-30.4%; p = 0.01) . Blood and/or blood components transfusion was independently associated with neonatal LO-BSI (OR: 21.2; 95% CI: 1.1-423) . CONCLUSIONS: LO-BSI infection prolongs hospital stay and is associated with increased mortality among neonates . In the present series, blood transfusion was a significant risk factor for LO-BSI. J Infect Dis, 2002 Jun 15, 185(12), 1754 - 60 Epub 2002 May 23. Humanlike immune response of human leukocyte antigen-DR3 transgenic mice to staphylococcal enterotoxins: a novel model for superantigen vaccines; DaSilva L et al.; This study examined the biologic responses of transgenic mice expressing human leukocyte antigen (HLA)-DR3 and human CD4 molecules, in the absence of murine major histocompatibility complex (MHC) class II molecules (Ab(0)), to staphylococcal enterotoxins (SEs) and evaluated protective immunity of a nonsuperantigen form of SEB against wild-type holotoxin . HLA-DR3 transgenic mice responded to several log lower concentrations of SEs and secreted higher levels of proinflammatory cytokines than did wild-type mice . Vaccination of transgenic mice with a nonsuperantigenic form of SEB induced high levels of neutralizing anti-SEB antibodies, which protected the mice from a surge in proinflammatory cytokine secretion after SEB challenge . The humanlike responses of the transgenic mice to SEs support the hypothesis that these mice represent an appropriate model to examine vaccines and therapeutics against SEs . This is thought to be the first report of examination of a vaccine against SEB in the context of human MHC class II receptors. Infez Med, 2001 Mar, 9(1), 13 - 8 {Vascular graft infection by Staphylococcus epidermidis: efficacy of various perioperative prophylaxis protocols in an animal model}; Giacometti A et al.; A rat model was used to investigate the efficacy of levofloxacin, cefazolin and teicoplanin in the prevention of vascular prosthetic graft infection . Graft infections were established in the subcutaneous tissue of 300 male Wistar rats by implantation of Dacron prostheses followed by topical inoculation with methicillin-susceptible and methicillin-resistant S . epidermidis . The study included a group without contamination, two contaminated groups without prophylaxis, two contaminated groups with intraperitoneal levofloxacin prophylaxis (10 mg/kg), two contaminated groups with intraperitoneal cefazolin prophylaxis (30 mg/kg), two contaminated groups with intraperitoneal teicoplanin prophylaxis (10 mg/kg) and six contaminated groups with rifampin-soaked graft and intraperitoneal levofloxacin, cefazolin or te- icoplanin prophylaxis . The grafts were removed after 7 days and evaluated by quantitative culture . The efficacy of levofloxacin against the methicillin- susceptible strain did not differ from that of cefazolin or teicoplanin . Levofloxacin showed slight less efficacy than teicoplanin against the methicillin-resistant strain . The levofloxacin-rifampin combination proved to be similarly effective to the rifampin-teicoplanin combination and more effective than the rifampin-cefazolin combination against both strains . The rifampin-levofloxacin combination may be useful for the prevention of late-appearing vascular graft infections caused by S . epidermidis because it takes advantage of the good anti-staphylococcal activity of both drugs. South Med J, 2002 Jun, 95(6), 650 - 2 Severe hepatitis associated with oxacillin therapy; Al-Homaidhi H et al.; A 6-year-old girl had fever, abdominal pain, and severe anicteric hepatitis during intravenous oxacillin therapy for staphylococcal osteomyelitis . She had greatly elevated liver enzymes, prolonged prothrombin time, leukopenia, and eosinophilia . Clinical symptoms resolved and laboratory data returned to normal after withdrawing oxacillin and substituting cefazolin . This hepatotoxicity appears to be specific to oxacillin and not to other beta-lactams . Monitoring liver function tests during oxacillin therapy, especially in patients receiving prolonged treatment, may be warranted. Ann Fr Anesth Reanim, 2002 May, 21(5), 392 - 8 {Therapeutic principles of staphylococcal infections--Role and limitations of standard compounds}; Domart Y; Antibiotic therapy plays an important (but not exclusive) role in the treatment of staphylococcal infections . Measures aimed at reducing the bacterial inoculum through local procedures must be envisaged as often as possible . The removal of any foreign, infected materials is essential to success . In this article, we review the different, active antibiotics available, their advantages and disadvantages and their indications . In the light of these data, we propose a therapeutic approach to severe bacterial infection caused by a cluster of Gram-positive cocci . Staphylococcal infections pose daily therapeutic problems, whether in open-care practice or intensive care units . The specificity of staphylococcal infections encountered in an intensive care setting require a therapeutic approach which takes account of the context, and particularly of the incidence of resistant staphylococcal infections. Immunogenetics, 2002 Jun, 54(3), 216 - 20 Epub 2002 Apr 20. Molecular characterization of bovine CD26 upregulated by a staphylococcal superantigen; Lee SU et al.; In this report, we describe the cloning, sequencing, and expression of the bovine orthologue of CD26 (BoCD26) . Several monoclonal antibodies specific for a molecule, activation molecule 3 (ACT3), aberrantly expressed on superantigen-stimulated bovine CD8(+) lymphocytes, reacted with recombinant BoCD26 expressed in COS-7 and CHO cells . We also showed that human CD8(+) T cells stimulated by a superantigen expressed CD26 at high levels . These results demonstrate that ACT3 is identical to BoCD26 and suggest that CD26 upregulation on CD8(+) T cells is a general phenomenon of superantigens and not limited to their effects on bovine cells. Protein Sci, 2002 Jul, 11(7), 1695 - 701 The dual role of a loop with low loop contact distance in folding and domain swapping; Linhananta A et al.; Alpha helices, beta strands, and loops are the basic building blocks of protein structure . The folding kinetics of alpha helices and beta strands have been investigated extensively . However, little is known about the formation of loops . Experimental studies show that for some proteins, the formation of a single loop is the rate-determining step for folding, whereas for others, a loop (or turn) can misfold to serve as the hinge loop region for domain-swapped species . Computer simulations of an all-atom model of fragment B of Staphylococcal protein A found that the formation of a single loop initiates the dominant folding pathway . On the other hand, the stability analysis of intermediates suggests that the same loop is a likely candidate to serve as a hinge loop for domain swapping . To interpret the simulation result, we developed a simple structural parameter: the loop contact distance (LCD), or the sequence distance of contacting residues between a loop and the rest of the protein . The parameter is applied to a number of other proteins, including SH3 domains and prion protein . The results suggest that a locally interacting loop (low LCD) can either promote folding or serve as the hinge region for domain swapping . Thus, there is an intimate connection between folding and domain swapping, a possible cause of misfolding and aggregation. J Clin Invest, 2002 Jun, 109(12), 1587 - 98 APRIL modulates B and T cell immunity; Stein JV et al.; The TNF-like ligands APRIL and BLyS are close relatives and share the capacity to bind the receptors TACI and BCMA . BLyS has been shown to play an important role in B cell homeostasis and autoimmunity, but the biological role of APRIL remains less well defined . Analysis of T cells revealed an activation-dependent increase in APRIL mRNA expression . We therefore generated mice expressing APRIL as a transgene in T cells . These mice appeared normal and showed no signs of B cell hyperplasia . Transgenic T cells revealed a greatly enhanced survival in vitro as well as enhanced survival of staphylococcal enterotoxin B-reactive CD4+ T cells in vivo, which both directly correlate with elevated Bcl-2 levels . Analysis of humoral responses to T cell-dependent antigens in the transgenic mice indicated that APRIL affects only IgM but not IgG responses . In contrast, T cell-independent type 2 (TI-2) humoral response was enhanced in APRIL transgenic mice . As TACI was previously reported to be indispensable for TI-2 antibody formation, these results suggest a role for APRIL/TACI interactions in the generation of this response . Taken together, our data indicate that APRIL is involved in the induction and/or maintenance of T and B cell responses. J Psychosom Res, 2002 Jun, 52(6), 501 - 9 A rating scale for fibromyalgia and chronic fatigue syndrome (the FibroFatigue scale); Zachrisson O et al.; OBJECTIVE: To construct an observer's rating scale sensitive to change for measuring severity and treatment outcome in fibromyalgia (FM) and chronic fatigue syndrome (CFS) patients . METHODS: A selection of items from the Comprehensive Psychopathological Rating Scale (CPRS) were repeatedly rated and used as outcome measure of a 24-week treatment study . In the study 100 women, fulfilling the criteria for both FM and CFS, received intermittent injections of a staphylococcus toxoid or placebo . Nine CPRS-items with high baseline incidence (cutoff 70%) were extracted and validated against global ratings and the Fibromyalgia Impact Questionnaire (FIQ) . The fibromyalgia and chronic fatigue syndrome rating scale (the FibroFatigue scale) was thereafter formed based upon the extracted items and three supplemented ones . The interrater reliability was tested in 27 consecutive patients of both sexes . RESULTS: The FibroFatigue scale is an observer's rating scale with 12 items measuring pain, muscular tension, fatigue, concentration difficulties, failing memory, irritability, sadness, sleep disturbances, and autonomic disturbances (items derived from the CPRS) and irritable bowel, headache, and subjective experience of infection (new items) . There was a statistically significant correlation between the CPRS-extracted items and global ratings as well as with the FIQ . The interrater reliability of the new scale was excellent (correlation coefficient.98), irrespective of the patients' gender . CONCLUSION: The FibroFatigue scale seems to be a reliable and valid measuring instrument with capacity to monitor symptom severity and change during treatment of FM/CFS patients. Mol Microbiol, 2002 Jun, 44(5), 1251 - 67 Protein engineering modulates the transport properties and ion selectivity of the pores formed by staphylococcal gamma-haemolysins in lipid membranes; Comai M et al.; Staphylococcal gamma-haemolysins are bicomponent toxins in a family including other leucocidins and alpha-toxin . Two active toxins are formed combining HlgA or HlgC with HlgB . Both open pores in lipid membranes with conductance, current voltage characteristics and stability similar to alpha-toxin, but different selectivity (cation instead of anion) . Structural analogies between gamma-haemolysins and alpha-toxin indicate the presence, at the pore entry, of a conserved region containing four positive charges in alpha-toxin, but either positive or negative in gamma-haemolysins . Four mutants were produced (HlgA D44K, HlgB D47K, HlgB D49K and HlgB D47K/D49K) converting those negative charges to positive in HlgA and HlgB . When all charges were positive, the pores had the same selectivity and conductance as alpha-toxin, suggesting that the cluster may form an entrance electrostatic filter . As mutated HlgC-HlgB pores were less affected, additional charges in the lumen of the pore were changed (HlgB E107Q, HlgB D121N, HlgB T136D and HlgA K108T) . Removing a negative charge from the lumen made the selectivity of both HlgA-HlgB D121N and HlgC-HlgB D121N more anionic . Residue D121 of HlgB is compensated by a positive residue (HlgA K108) in the HlgA-HlgB pore, but isolated in the more cation-selective HlgC-HlgB pore . Interestingly, the pore formed by HlgA K108T-HlgB, in which the positive charge of HlgA was removed, was as cation selective as HlgC-HlgB . Meanwhile, the pore formed by HlgA K108T-HlgB D121N, in which the two charge changes compensated, retrieved the properties of wild-type HlgA-HlgB . We conclude that the conductance and selectivity of the gamma-haemolysin pores depend substantially on the presence and location of charged residues in the channel. Clin Exp Immunol, 2002 Jun, 128(3), 532 - 7 Increased expression of interleukin-13 but not interleukin-4 in CD4+ cells from patients with the hyper-IgE syndrome; Gudmundsson KO et al.; Hyper IgE syndrome (HIES) is a rare immunodeficiency disorder characterized mainly by high levels of polyclonal IgE in serum and recurrent staphylococcal abscesses of the skin and lungs . The raised IgE levels have led researchers to study the synthesis of cytokines that regulate switching of immunoglobulin production towards IgE such as interleukin-4 (IL-4), IL-12 and interferon-gamma (IFN)-gamma . However, the role of IL-13 in the disease pathogenesis has not been investigated extensively . In this study, we investigated intracellular expression of IL-4 and IL-13 in mononuclear cells and CD4+ cells isolated from patients with HIES and healthy controls . Cells were stained intracellularly with antibodies directed against IL-4 and IL-13 and analysed by flow cytometry before and after activation with PMA and calcium ionophore . The mean proportion of resting or activated IL-4 and IL-13 expressing mononuclear cells were comparable in the two groups as well as the proportion of IL-4 expressing CD4+ cells . In contrast, the mean proportion of IL-13 expressing CD4+ cells was increased significantly in patients with HIES in both the resting and the activated state compared to healthy controls . We conclude that increased expression of IL-13 in CD4+ cells from patients with HIES could account, at least partly, for raised IgE levels in those individuals. J Pharm Biomed Anal, 2002 Jun 20, 29(1-2), 255 - 62 Levels of specific antibodies towards the major antigenic determinant of slime-producing Staphylococcus epidermidis determined by an enzyme immunoassay and their protective effect in experimental keratitis; Georgakopoulos CD et al.; Staphylococcus epidermidis is an important cause of bacterial keratitis . Certain S . epidermidis strains produce an extracellular slime layer rich in an acidic polysaccharide with a molecular size of 20 kDa (20-kDa PS) . We have demonstrated that the level of 20-kDa PS-specific antibodies significantly rises after establishment of slime-producing S . epidermidis bacteraemia and, furthermore, that rabbit polyclonal antibodies to 20-kDa PS opsonize cells of slime-producing S . epidermidis to a great degree and promote their clearance by polymorphonuclear cells (Arch . Biochem . Biophys . 342 (1997) 389; J . Pharm . Biomed . Anal . 22 (2000) 1029) . The purpose of this study was to examine the protective and therapeutic effects both of active immunization, using 20-kDa PS as antigen, and of passive administration of specific antibodies towards the 20-kDa PS in a rabbit keratitis model . For active immunization, 20 rabbits were subcutaneously immunized with 20-kDa PS, whereas for passive immunization specific polyclonal IgG antibodies against 20-kDa PS were administered to 20 rabbits 1 day before induction of infection . Clinical observations were made weekly for 1 month and levels of 20-kDa PS antibodies in serum and aqueous humor in both immunization groups were determined by an enzyme immunoassay . The levels of specific anti-20-kDa PS IgG in serum and aqueous humor following either active or passive immunization were significantly higher as compared with control groups (P<0.001) . Although, actively immunized rabbits showed significantly less corneal damage than control animals, passively immunized ones were significantly better protected as compared with both control and those actively immunized . Obtained results suggest that 20-kDa PS plays crucial role in the pathogenesis of S . epidermidis keratitis and that both types of immunization significantly protect against corneal S . epidermidis pathology and damage. J Biomol NMR, 2002 May, 23(1), 47 - 55 Angular dependence of 1J(Ni,Calphai) and 2J(Ni,Calpha(i-1)) coupling constants measured in J-modulated HSQCs; Wirmer J et al.; A new method to measure 1J(Ni,Calphai) and 2J(Ni,Calpha(i-1)) coupling constants in proteins based on a J-modulated sensitivity enhanced HSQC was introduced . Coupling constants were measured in the denatured and in the native state of ubiquitin and found to depend on the conformation of the protein backbone . Using a combined data set of experimental coupling constants from ubiquitin and staphylococcal nuclease (Delaglio et al., 1991), the angular dependence of the coupling constants on the backbone angles psi and phi was investigated . It was found that the size of 2J(Ni,Calpha(i-1)) correlates strongly with the backbone conformation, while only a weak conformational dependence on the size of 1J(Ni,Calphai) coupling constants was observed . Coupling constants in the denatured state of ubiquitin were uniform along the sequence of the protein and not dependent on a given residue type . Furthermore it was shown that the observed coupling constants were in good agreement with predicted coupling constants using a simple model for the random coil. Vnitr Lek, 2002 Apr, 48(4), 302 - 6 {Pathogen resistance and other risk factors in the frequency of lower limb amputations in patients with the diabetic foot syndrome}; Fejfarova V et al.; Patients with diabetes mellitus undergo more amputations due to peripheral vascular disease, neuropathy and especially to infection requiring long-lasting antibiotic therapy than non diabetic patients . The aim of our study was to assess the association between the presence of resistant pathogens presented in diabetic ulcers and the frequency of lower limb amputations . METHODS: 191 diabetic patients consecutively treated for the diabetic foot in our foot clinic were included into two years retrospective study . Peripheral ischemia, the presence of osteomyelitis and the incidence of all Gram positive and negative resistant pathogens (defined as resistance to all oral antibiotics) especially of resistant Staphylococcus species presenting in diabetic foot ulcers were determined . RESULTS: 50/191 (26%) patients underwent amputation, of whom 44/50 (88%) had minor and 6/50 (12%) had major amputations . 53/181 (29%) patients with diabetic foot ulcers had resistant pathogens in their defects . Amputated patients had significantly more resistant microorganisms than patients without amputations--24/42 (57%) vs . 29/139 (21%); p < 0.001 . Resistant Staphylococcus species were found in 21% (38/181) of all patients . Patients with amputations had significantly more resistant Staphylococcus species in comparison with patients without amputations--18/42 (43%) vs . 20/139 (14%); p < 0.001 . Significantly higher incidence of peripheral vascular disease--79% (38/48) vs . 60% (81/136); p < 0.05 and osteomyelitis--69% (33/48) vs . 13% (18/140); p < 0.001--were found in patients with amputations in comparison with patients without amputations . CONCLUSION: The presence of pathogens resistant to all oral antibiotics and especially of resistant Staphylococcus species was significantly higher in diabetic patients with lower limb amputations in comparison with patients without amputations. Proc Natl Acad Sci U S A, 2002 Jun 11, 99(12), 8289 - 94 A critical concentration of neutrophils is required for effective bacterial killing in suspension; Li Y et al.; We have examined the effect of neutrophil concentration on killing of a clinical isolate of Staphylococcus epidermidis . Human neutrophils at concentrations varying from 10(5) to 10(7) per ml were mixed in suspension with S . epidermidis at concentrations varying from 10(3) to 10(8) colony-forming units/ml, and the concentration of viable bacteria was assayed after various times at 37 degrees C . The rate of bacterial killing depended on the concentration of neutrophils and not on the ratio of neutrophils to bacteria . Below a critical concentration of neutrophils, bacteria growth was greater than neutrophil killing of bacteria even when the ratio of neutrophils to bacteria was 100:1 . We fitted the time course of bacterial concentration and its dependence on neutrophil concentration with an exponential function, the exponent of which is (-kp + g)t, where k is the second-order rate constant for bacterial killing, p is the neutrophil concentration, g is the first-order rate constant for bacterial growth, and t is time . We found that k approximately 2 x 10(-8) ml per neutrophil per min, and g approximately 8 x 10(-3)/min . Only when p is greater than g/k, which we call the critical neutrophil concentration, does the bacterial concentration fall . Under optimal assay conditions, the critical neutrophil concentration was 3-4 x 10(5) per ml, a value very close to that (< or =5 x 10(5) per ml) known to predispose humans to bacterial and fungal infections. J Interferon Cytokine Res, 2002 May, 22(5), 593 - 601 The combination of soluble IL-18Ralpha and IL-18Rbeta chains inhibits IL-18-induced IFN-gamma; Reznikov LL et al.; Although the beta chain of interleukin-18 receptor (IL-18Rbeta) is required for signaling, the soluble (extracellular) form does not bind IL-18, and its role in inhibiting IL-18 is unclear . In the present study, both the soluble human IL-18 ligand binding alpha chain (sIL-18Ralpha) and the sIL-18Rbeta chain were investigated for inhibition of IL-18-induced interferon-gamma (IFN-gamma) production in human peripheral blood mononuclear cells (PBMC), whole blood, and KG-1 macrophage and natural killer (NK) cell lines . Neutralization of IL-18 by soluble receptors was compared with that of the IL-18 binding protein (IL-18BP) . An equimolar concentration IL-18BP inhibited 90% of IL-18 activity, whereas a 4-fold molar excess of sIL-18Ralpha had no effect . A dimeric construct of sIL-18Ralpha linked to the Fc domain of IgG1 (sIL-18Ralpha:Fc) increased IL-18 activity 2.5-fold . In PBMC stimulated with lypopolysaccharide (LPS) or in whole blood stimulated with Staphylococcus epidermidis, 3 nM IL-18BP reduced IFN-gamma by 80%, whereas IL-18Ralpha:Fc had no effect . A construct of the sIL-18Rbeta linked to Fc (sIL-18Rbeta:Fc) did not affect IL-18-induced IFN-gamma even at 80-fold molar excess of IL-18 . However, the combination of both soluble receptors reduced IFN-gamma by 80% . In KG-1 cells, a 50% reduction in IL-18 activity was observed using an 80-fold molar excess of sIL-18Ralpha:Fc but only in the presence of sIL-18Rbeta:Fc . Similarly, a 50% reduction was observed using sIL-18Rbeta:Fc in the presence of a molar excess of sIL-18Ralpha:Fc . Similar inhibition was observed in NK cells . These studies reveal that the combination of the ligand-binding and the nonligand-binding extracellular domains of IL-18R is needed to inhibit IL-18, whereas IL-18BP neutralizes at equimolar concentration. Magy Onkol, 2000 Dec 1, 44(4), 289 - 295 {Frequent infections of neutropenic pediatric patients and therapeutic modalities}; Muller J et al.; Neutropenia, resulting from intensive chemotherapy is a common problem . The appearance of fever in neutropenic patients should always raise the suspicion of infection and should be followed by an intensive diagnostic evaluation and start of antibacterial treatment . The authors analyzed the association between isolated bacteria from blood cultures and the clinical background of all febrile episodes that occurred in neutropenic children in a two-year long period . Comparable to the international trends, our results suggest an increased prevalence of the Gram-positive organisms causing bacteriaemia . The clear majority of the isolated bacteria was coagulase-negativ Staphylococcus (cnS), which is a multiresistant strain, and sensitive only to the glycopeptide antibiotics . This latter fact can be a consequence of the frequent use of central venous catheters . The empirical therapy, the therapy used in microbiologically and clinically proved infections, and the supplementary and prophylactic methods of treatment are presented. Pediatr Dermatol, 2002 May-Jun, 19(3), 220 - 3 Recurring staphylococcal scalded skin syndrome-like bullous mastocytosis: the utility of cytodiagnosis and the rapid regression with steroids; Has C et al.; We report a male infant with onset of an extensive bullous eruption at the age of 45 days . Staphylococcal scalded skin syndrome (SSSS) was suspected . Bullous mastocytosis was diagnosed by cytodiagnosis and confirmed by histologic examination . Three serious relapses were noted in a 2-year follow-up, and SSSS was again suspected because of high fever and leukocytosis with neutrophilia in an infectious context . Cytodiagnosis revealed the presence of mast cells and permitted rapid diagnosis of recurrences of bullous mastocytosis . Systemic corticotherapy dramatically improved the cutaneous lesions and general symptoms . This case report emphasizes the utility of cytodiagnosis in extensive blistering diseases in infancy and the possibility of obtaining rapid healing by using steroids. Eur J Biochem, 2002 Jun, 269(11), 2647 - 55 Human immunoglobulin A (IgA)-specific ligands from combinatorial engineering of protein A; Ronnmark J et al.; Affinity reagents capable of selective recognition of the different human immunoglobulin isotypes are important detection and purification tools in biotechnology . Here we describe the development and characterization of affinity proteins (affibodies) showing selective binding to human IgA . From protein libraries constructed by combinatorial mutagenesis of a 58-amino-acid, three-helix bundle domain derived from the IgG-binding staphylococcal protein A, variants showing IgA binding were selected by using phage display technology and IgA monoclonal antibodies (myeloma) as target molecules . Characterization of selected clones by biosensor technology showed that five out of eight investigated affibody variants were capable of IgA binding, with dissociation constants (K(d)) in the range between 0.5 and 3 microm . One variant (Z(IgA1)) showing the strongest binding affinity was further analyzed, and showed that human IgA subclasses (IgA(1) and IgA(2)) as well as secretory IgA were recognized with similar efficiencies . No detectable cross-reactivity towards human IgG, IgM, IgD or IgE was observed . The potential use of the Z(IgA1) affibody as a ligand in affinity chromatography applications was first demonstrated by selective recovery of IgA protein from a spiked Escherichia coli total cell lysate, using an affinity column containing a divalent head-to-tail Z(IgA1) affibody dimer construct as a ligand . In addition, efficient affinity recovery of IgA from unconditioned human plasma was also demonstrated. Clin Nucl Med, 2002 Jun, 27(6), 401 - 4 Ga-67 SPECT to detect endocarditis after replacement of an aortic valve; Pena FJ et al.; Ga-67 SPECT was crucial in making the diagnosis of infectious endocarditis in a 28-year-old man with a history of aortic valve replacement who was referred to the nuclear medicine service because of persistent fever . In the planar left anterior oblique view, an area of slight, diffuse activity was noted . The shape and site of the infectious focus were well defined by Ga-67 SPECT . After Ga-67 imaging, results of a computed tomographic (CT) scan were negative and the patient was treated with antibiotics and discharged from the hospital after showing clinical improvement . However, the fever returned and he was readmitted . A second Ga-67 SPECT was also positive, but a spiral CT scan showed only minor inflammatory changes . Finally, transesophageal ultrasonographic images suggested vegetation on the prosthetic valve . Cultures of the graft and the valve were obtained surgically and showed staphylococcal endocarditis . Exaggerated confidence in negative CT scan results led to unnecessarily delayed surgical treatment of the infection . Ga-67 SPECT was a powerful noninvasive tool in the diagnosis of infectious complications after prosthetic valve surgery. J Neuroimmunol, 2002 Jun, 127(1-2), 44 - 53 Involvement of noradrenergic nerves in the activation and clonal deletion of T cells stimulated by superantigen in vivo; del Rey A et al.; Superantigens, like staphylococcal enterotoxin B (SEB), induce a strong proliferative response followed by clonal deletion of a substantial portion of defined Vbeta T cells . The remaining cells display in vitro anergy . We found that the immune response to SEB was paralleled by biphasic changes in the activity of the sympathetic nervous system . Furthermore, sympathetic denervation resulted in decreased SEB-induced cell proliferation and IL-2 production, and impeded the specific deletion of splenic CD4Vbeta8 cells observed in intact animals without affecting anergy . These studies provide the first evidence of an immunoregulatory cross-talk between sympathetic nerves and superantigen-activated immune cells. FEMS Microbiol Lett, 2002 May 7, 210(2), 263 - 70 General expression vectors for Staphylococcus carnosus enabled efficient production of the outer membrane protein A of Klebsiella pneumoniae; Hansson M et al.; General expression vectors, designed for intracellular expression or secretion of recombinant proteins in the non-pathogenic Staphylococcus carnosus, were constructed . Both vector systems encode two different affinity tags, an upstream albumin binding protein and a downstream hexahistidyl peptide, and are furnished with cleavage sites for two site-specific proteases for optional affinity tag removal . To evaluate the novel vectors, the gene encoding the outer membrane protein A (OmpA) of Klebsiella pneumoniae was introduced into the vectors . Efficient production was demonstrated in both systems, although, as expected for OmpA fusions, somewhat better intracellularly, and the fusion proteins could be recovered as full-length products by affinity chromatography. FEMS Microbiol Lett, 2002 May 7, 210(2), 209 - 14 Decarboxylase activity involved in methyl ketone production by Staphylococcus carnosus 833, a strain used in sausage fermentation; Fadda S et al.; Staphylococcus carnosus strain 833, inoculated into sausage, increased the levels of methyl ketones which contributed to the cured aroma . These ketones were predicted to arise from incomplete beta-oxidation followed by a decarboxylation . To check this hypothesis, we measured the beta-decarboxylase activity in resting cells of S . carnosus grown in complex or in synthetic medium, using as substrate a beta-ketoacid, which can be an intermediate of the beta-oxidation pathway . This activity was present throughout the growth period . The enzyme appeared to be constitutive because no induction was observed . High aeration, a pH of 5 and the presence of nitrate promoted the production of methyl ketones. Klin Med (Mosk), 2002, 80(4), 58 - 61 {Staphylococcal toxic shock syndrome in young women}; Novikov IuI et al.; A literature review is presented on staphylococcal toxic shock syndrome . A case of this syndrome in a 39-year-old woman is reported . Clinical, pathology, treatment data on this condition are analysed. J Chemother, 1991 Jan, 3 Suppl 1, 163 - 5 Epidemiological surveillance of staphylococcal resistance; Giacometti A et al.; The aim of our investigation was to characterize better the possible changes in staphylococcal susceptibility to some antimicrobial agents . We considered the staphylococcal strains isolated between June 1986 and June 1989 and held responsible for proved bacterial infections . In this relatively short time we observed a moderate increase in methicillin resistance . Compared with methicillin-sensitive strains, methicillin-resistant isolates showed minor susceptibility to most antimicrobial agents; nevertheless, vancomycin, teicoplanin, cefamandole, imipenem, fluoroquinolones and netilmicin, according to our results, appear at present the most reliable antistaphylococcal antibiotics. Przegl Lek, 2001, 58(12), 1029 - 33 {Selected parameters of the cellular and humoral immunity in atopic dermatitis . Relationship to the severity of the disease}; Adamek-Guzik T et al.; Despite the great progress, our understanding of the pathogenesis of atopic dermatitis (AD) is still incomplete . In particular, the clinical importance of various changes of the immune system parameters is unclear . Accordingly we have undertaken the study to compare selected parameters of cellular and humoral immunity between AD subjects (n = 26) and healthy controls (n = 10) . These parameters included immunoglobulin levels (IgE in particular), neutrophil respiratory, oxygen burst, peripheral blood lymphocyte phenotype and response to mitogens . We also analysed the relationship between these parameters and clinical severity of skin lesions . RESULTS: Mean total immunoglobulin E levels were very significantly increased in the AD group (1563 +/- 459 vs 35.5 +/- 12.1 IU/ml; p = 0.001) . Simultaneously total serum IgE levels varied extensively between individual subjects with AD and were significantly correlated to clinical severity of the disease (Rs = 0.44; p = 0.02) . Atopic dermatitis was also associated with the increase in the number of CD4+ and simultaneous decrease in the CD8+ lymphocytes causing statistically significant difference in CD4:CD8 ratio compared to the control group . We also observed changes of proliferation indices to phytohaemagglutinin (decrease) and increase of responses to anti CD3 mAb (OKT-3) and staphylococcal enterotoxin B . None of these immune parameters however, appeared to be statistically correlated to clinical status . CONCLUSIONS: We find that atopic dermatitis is associated with significant changes of several important indices of cellular and humoral immunity including increased IgE levels and altered peripheral lymphocyte proliferation capacity and phenotype . Change of total IgE levels appears to be the most important from clinical point of view. J Clin Microbiol, 2002 Jun, 40(6), 2275 - 7 Population diversity of Staphylococcus intermedius isolates from various host species: typing by 16S-23S intergenic ribosomal DNA spacer polymorphism analysis; Bes M et al.; Twelve 16S-23S ribosomal DNA intergenic spacer (ITS-PCR) types were identified among 57 Staphylococcus intermedius isolates from humans and other animals . Six ITS-PCR types were host specific, and most human and canine strains belonged to the same types (A and J) . Pigeon, horse, and mink strains appeared more heterogeneous. Ukr Biokhim Zh, 2001 Jul-Aug, 73(4), 24 - 8 {Effect of staphylococcus active substances on ATPase activity of smooth muscle actomyosin and myosin}; Davydovs'ka TL et al.; The effect of staphylococcus active substances--protein A (PA) and peptidoglican (PG) at concentrations 10(-6)-10(-2) mg/ml on the ATPase activity of pig stomach natural actomyosin and myosin was studied . It was shown that PA and PG at direct contact with smooth muscle contractile proteins caused the activation and inhibition of ATPase activity, respectively . On the basis of this investigation it was assumed that staphylococcal active substances were able to modify of the ATPase activity smooth muscle contractile proteins perhaps via direct action on the myosin molecule, which could be accompanied by conformational changes of the active center of myosin ATPase. Protein Eng, 2002 May, 15(5), 383 - 91 Does fusion of domains from unrelated proteins affect their folding pathways and the structural changes involved in their function? A case study with the diphtheria toxin T domain; Chenal A et al.; We investigated whether the structural and functional behaviors of two unrelated protein domains were modified when fused . The IgG-binding protein ZZ derived from staphylococcal protein A was fused to the N- and/or C-terminus of the diphtheria toxin transmembrane domain (T) . T undergoes a conformational change from a soluble native state at neutral pH to a molten globule-like state at acidic pH, leading to its interaction with membranes . We found that this molten globule state was not connected to the GdnHCl-induced unfolding pathway of T . The pH-induced transition of T, and also the unfolding of T and ZZ at neutral and acidic pH, were unchanged whether the domains were isolated or fused . The position of ZZ, however, influenced the solubility of T near its pK(i) . SPR measurements revealed that T has a high affinity for membranes, isolated or within the fusion proteins (K(D)< 10(-11) M) . This work shows that in the case of T and ZZ, the fusion of protein domains with different stabilities does not alter the structural changes involved in folding and function . This supports the use of T as a soluble membrane anchor. Vet Microbiol, 2002 Jun 20, 87(2), 175 - 82 Identification and antimicrobial susceptibility of Staphylococcus chromogenes isolates from intramammary infections of dairy cows; Devriese LA et al.; Staphylococcus chromogenes is a highly prevalent species in subclinical mastitis with a well-established impact on somatic cell count . Few data are available on its antimicrobial susceptibility . The objective of this study was three-fold: (1) to evaluate simple identification tests by comparing them with a genomic method; (2) to determine minimal inhibitory concentrations (MICs) of different antibiotics; (3) to search for the presence of important resistance mechanisms and resistance-determining genes.Seventy-three staphylococcal strains, all collected on different dairy farms, were tentatively identified as S . chromogenes based on their lack of hemolysis and their characteristic intermediate DNase activity . The identification of 70 strains was confirmed as S . chromogenes by tRNA intergenic spacer PCR (tRNA PCR) . Three strains were identified as S . sciuri, a species that is naturally cloxacillin- and lincomycin-resistant.All 70 S . chromogenes strains were found to be normally susceptible to neomycin, gentamicin, erythromycin, enrofloxacin, and to penicillinase-stable penicillins and cephalosporins, represented in this study by cloxacillin . The latter result was confirmed by the absence of the mecA gene in each of 13 strains in which this gene was searched for . Twenty-seven (38%) strains were penicillinase producers . Three lincomycin-resistant S . chromogenes strains were found to carry the linA gene.It was concluded that S . chromogenes can be identified reliably in routine mastitis bacteriology, and that the only resistance of importance is against penicillinase-susceptible penicillins. J Am Chem Soc, 2002 Jun 5, 124(22), 6378 - 82 Avidin: a natural bridge for quantum dot-antibody conjugates; Goldman ER et al.; We describe the preparation and characterization of bioinorganic conjugates in which luminescent semiconductor CdSe-ZnS core-shell nanocrystal quantum dots (QDs) were coupled to antibodies through the use of an avidin bridge adsorbed to the nanocrystal surface via electrostatic self-assembly . Avidin, a highly positively charged protein, was found to adsorb tightly to QDs modified with dihydrolipoic acid, which gives their surface a homogeneous negative charge . QD conjugation to biotinylated antibodies subsequently is readily achieved . Fluoroimmunoassays utilizing these antibody conjugated QDs were successful in the detection of protein toxins (staphylococcal enterotoxin B, cholera toxin) . QD-antibody conjugates formed in such a facile manner permit their use as a common immuno reagent, and in the development of multianalyte detection. Pol Arch Med Wewn, 2001 Nov, 106(5), 1055 - 8 {Diagnostic and therapeutic problem of infected arteriovenous fistula for hemodialysis-case report}; Kade G et al.; On the case of 48 old patient with chronic renal failure, treated with repeated hemodialyses, we described diagnostic and therapeutic problems of infected arterio-venous fistula of severe clinical course and rare epidemiology (Staphylococcus warneri) . Special attention was paid to the difficulties in the confirmation of bacterial etiology of this infection as well as practical usefulness of ultrasonography in the monitoring of clinical course and therapy of infected arteriovenous fistula. J Int Med Res, 2002 Mar-Apr, 30(2), 99 - 108 Differential cytokine response in host defence mechanisms triggered by gram-negative and gram-positive bacteria, and the roles of gabexate mesilate, a synthetic protease inhibitor; Iwadou H et al.; Bacterial infection results in the production of inflammatory mediators and may be involved in the pathogenesis of sepsis and/or systemic inflammatory response syndrome . The effect of lipopolysaccharide (LPS), a major component of the outer surface of Gram-negative bacteria, and Staphylococcal enterotoxin B (SEB), a superantigen of Gram-positive bacteria, on cytokine production in peripheral blood mononuclear cells (PBMCs) was examined . LPS significantly increased the production of proinflammatory and anti-inflammatory cytokines, and SEB enhanced the production of helper T lymphocyte type cytokines . These results illustrated the different responses to Gram-negative and Gram-positive bacterial infections . The effect of gabexate mesilate, a synthetic protease inhibitor, on cytokine production and expression of the toll-like receptor (TLR) was also examined . The results suggest that gabexate mesilate-induced inhibition of tumour necrosis factor-alpha (TNF-alpha) and interleukin-18 (IL-18) production in LPS-stimulated PBMCs is due to the inhibition of the nuclear factor-kappa B activation pathway and/or inhibition of the processing pathway of pro-TNF-alpha and pro-IL-18, not to down-regulation of TLR-2 or TLR-4. Indian J Med Res, 2001 Nov, 114, 169 - 72 A simple alternative method for rapid detection of slime produced by Staphylococcus epidermidis isolates in bacterial keratitis; Nayak N et al.; BACKGROUND & OBJECTIVES: Slime is a known virulence factor of Staphylococcus epidermidis . The conventional Christensen's method for detection of slime in the laboratory takes at least 48 h . We, therefore, tried to evaluate the efficacy of the Congo red agar method as a routine procedure for detecting slime among isolates from corneal ulcers . METHODS: A total of 244 isolates from corneal ulcers were identified as S . epidermidis by the standard procedures . Slime was detected both by the conventional Christensen's method as well as by the Congo red agar method . RESULTS: Ninety two (37.7%) isolates were positive and 86 (35.2%) were negative for slime by both the techniques . Fifty four (22.1%) isolates were positive in Congo red agar, but negative by Christensen's method; whereas only 12 (4.9%) were negative by Congo red but positive by Christensen's method . Detection of slime by Congo red agar method was rapid i.e., all the 146 strains were positive within 24 h of incubation . On the other hand, Christensen's method had a delayed response; 42.3 per cent (44/104) strains being negative during the first 24 h of incubation . INTERPRETATION & CONCLUSION: Our results suggested that culture on Congo red agar was a sensitive and rapid test for detecting slime . This might help in the quick identification in a routine laboratory of slime positive isolates in bacterial keratitis. Indian J Med Res, 2001 Nov, 114, 160 - 3 Comparative evaluation of hydrophobicity measures for virulence determination of Staphylococcus epidermidis from hospitalized patients & healthy individuals; Das SC et al.; BACKGROUND & OBJECTIVES: Hydrophobicity is one of the recognized markers for identifying pathogenic strains of Staphylococcus epidermidis . A number of tests are available for measuring the hydrophobic character but three tests viz., salt aggregation test (SAT), n-hexadecane adherence assay (HAA) and xylene adherence assay (XAA) are in common practice with different degrees of sensitivity . However, in the Indian context, no systematic study has been reported on the hydrophobicity of S . epidermidis . Hence, the present study was undertaken to compare the three methods for hydrophobicity measurement for identifying the pathogenic isolates of S . epidermidis . METHODS: Of the 597 samples obtained from milker's and butcher's (hand and nose), hospitalized pre-operative patients (hand, nose, ear), and post-operative patients (blood) examined, 212 isolates of S . epidermidis were recovered using established laboratory procedures . The isolates were screened by the three tests viz., SAT, HAA and XAA . The identified hydrophobic isolates were further tested by mouse inoculation method . RESULTS: Of the 212 S . epidermidis isolates studied, 24 (11.32%), 23 (10.84%) and 4 (1.88%) were found to be hydrophobic as detected by HAA, XAA and SAT, respectively . No isolates from apparently healthy individuals were found to be hydrophobic by SAT while 13 (8.76%) and 10 (8.24%) strains from apparently healthy individuals were detected to be hydrophobic by HAA and XAA, respectively . Seven (33.38%) isolates each were hydrophobic by HAA and XAA and 4 (22.22%) by SAT among the isolates from hospitalized post-operative patients . INTERPRETATION & CONCLUSION: The results suggested that there was no difference in the number of strains detected as hydrophobic by HAA and XAA, and any of the two tests may be used for screening the hydrophobic strains of S . epidermidis from healthy individuals and patients. Biophys J, 2002 Jun, 82(6), 3289 - 304 Experimental pK(a) values of buried residues: analysis with continuum methods and role of water penetration; Fitch CA et al.; Lys-66 and Glu-66, buried in the hydrophobic interior of staphylococcal nuclease by mutagenesis, titrate with pK(a) values of 5.7 and 8.8, respectively (Dwyer et al., Biophys . J . 79:1610-1620; Garcia-Moreno E . et al., Biophys . Chem . 64:211-224) . Continuum calculations with static structures reproduced the pK(a) values when the protein interior was treated with a dielectric constant (epsilon(in)) of 10 . This high apparent polarizability can be rationalized in the case of Glu-66 in terms of internal water molecules, visible in crystallographic structures, hydrogen bonded to Glu-66 . The water molecules are absent in structures with Lys-66; the high polarizability cannot be reconciled with the hydrophobic environment surrounding Lys-66 . Equilibrium thermodynamic experiments showed that the Lys-66 mutant remained folded and native-like after ionization of the buried lysine . The high polarizability must therefore reflect water penetration, minor local structural rearrangement, or both . When in pK(a) calculations with continuum methods, the internal water molecules were treated explicitly, and allowed to relax in the field of the buried charged group, the pK(a) values of buried residues were reproduced with epsilon(in) in the range 4-5 . The calculations show that internal waters can modulate pK(a) values of buried residues effectively, and they support the hypothesis that the buried Lys-66 is in contact with internal waters even though these are not seen crystallographically . When only the one or two innermost water molecules were treated explicitly, epsilon(in) of 5-7 reproduced the pK(a) values . These values of epsilon(in) > 4 imply that some conformational reorganization occurs concomitant with the ionization of the buried groups. Immunopharmacol Immunotoxicol, 2002 Feb, 24(1), 113 - 9 The polyphenol chlorogenic acid inhibits staphylococcal exotoxin-induced inflammatory cytokines and chemokines; Krakauer T; Proinflammatory cytokines mediate the toxic effect of staphylococcal exotoxins (SE) . Chlorogenic acid, a plant polyphenol, inhibited SE-induced T-cell proliferation (by 98%) and production of interleukin 1beta, tumor necrosis factor, interleukin 6, interferon gamma, monocyte chemotactic protein I (MCP-l), macrophage inflammatory protein (MIP)-lalpha, and MIP-lbeta by human peripheral blood mononuclear cells . These data indicate that chlorogenic acid may be therapeutically useful for mitigating the pathogenic effects of SE . Naturally occurring polyphenolic compounds such as chlorogenic acid may serve as a potent anti-inflammatory agent alternative to conventional chemotherapeutics. J Surg Res, 2002 May 15, 104(2), 124 - 30 Toll-like receptor-4 signaling mediates pulmonary neutrophil sequestration in response to gram-positive bacterial enterotoxin; Calkins CM et al.; BACKGROUND: Toll-like receptors (TLRs) serve as mediators of innate immune responses to pathogen-associated molecular patterns (PAMPs) which include lipopolysaccharide (LPS) and staphylococcal enterotoxin B (SEB) . TLR-4 is thought to act as the primary effector of LPS recognition and TLR-2 is thought to mediate responses to Gram-positive bacterial proteins . Chemokines such as macrophage inflammatory protein (MIP-2) are peptides that are responsible for lung neutrophil (PMN) sequestration following an infectious or inflammatory insult . Given the Gram-positive origin of SEB, we hypothesized that mice with altered TLR-4 signaling would exhibit no difference in lung PMN sequestration following SEB when compared to wild-type mice . METHODS: Wild-type and TLR-4 mutant mice were administered intratracheal saline, LPS (Escherichia coli 0.1 mg/kg), or SEB (1 mg/kg) . After 24 h, lung PMN accumulation was determined by myeloperoxidase (MPO) assay and bronchoalveolar lavage fluid cell count (BALfcc) . Total lung and BALf MIP-2 was measured by enzyme-linked immunosorbent assay . RESULTS: There was an increase in lung PMN accumulation (by both MPO and BALfcc) and MIP-2 following LPS and SEB in wild-type mice compared to saline-treated controls . In contrast, TLR-4 mice failed to exhibit an increase in lung MIP-2 or PMN accumulation following either LPS or SEB compared to wild-type mice . CONCLUSIONS: TLR-4 mutant mice are unresponsive to intratracheal LPS . SEB elicited an increase in lung MIP-2 and PMN accumulation in wild-type mice . However, TLR-4 mutant mice were protected from this process . This suggests that TLR-4 signaling may mediate the responses to other PAMPs in addition to LPS . (c) 2002 Elsevier Science (USA). Anticancer Res, 2002 Mar-Apr, 22(2A), 769 - 76 T-cell immunotherapy for human MK-1-expressing tumors using a fusion protein of the superantigen SEA and anti-MK-1 scFv antibody; Ueno A et al.; BACKGROUND: The bacterial superantigen staphylococcal enterotoxin A (SEA) is an extremely potent activator of T lymphocytes when presented on major histocompatibility complex (MHC) class II molecules . To develop a tumor-specific superantigen for cancer therapy, we constructed a recombinant fusion protein of SEA and the single-chain variable fragment (scFv) of the FU-MK-1 antibody, which recognizes a glycoprotein antigen (termed MK-1 antigen) present on most carcinomas . MATERIALS AND METHODS: We employed recombinant DNA techniques to fuse recombinant mutant SEA to an scFv antibody derived from FU-MK-1 and the resulting fusion protein (SEA/FUscFv) was produced by a bacterial expression system, purified with a metal-affinity column, and characterized for its MK-1-binding specificity and its antitumor activity . RESULTS: The SEA/FUscFv fusion protein retained the reactivity with MK-1-expressing tumor cells, introduced a specific cytotoxicity of lymphokine-activated killer T-cells to the tumor cells, and consequently suppressed the tumor growth in a SCID mouse xenograft model . CONCLUSION: This genetically engineered SEA/FUscFv fusion protein may serve as a potentially useful immunotherapeutic reagent for human MK-1-expressing tumors. Infect Immun, 2002 Jun, 70(6), 2989 - 94 Pirfenidone blocks the in vitro and in vivo effects of staphylococcal enterotoxin B; Hale ML et al.; Pirfenidone {5-methyl-1-phenyl-2-(1H)-pyridone} down-regulates expression of cytokines and other mediators involved in the onset and development of pulmonary fibrosis . Pirfenidone also inhibits production of tumor necrosis factor alpha (TNF-alpha) from macrophages incubated with endotoxin and protects mice against endotoxin shock . Pirfenidone's ability to reduce cytokine expression in these disorders led us to investigate the drug's effect on another cytokine anomaly, superantigen-induced shock . BALB/c mice were exposed to staphylococcal enterotoxin B (SEB) either systemically or by aerosol and subsequently potentiated with a sublethal dose of lipopolysaccharide . In these experiments, pirfenidone given 2 to 4.25 h after SEB resulted in 80 to 100% survival versus only 0 to 10% survival among untreated control animals . Relative to serum cytokine levels from controls given toxin but no drug, there was a 35 to 80% decrease in TNF-alpha, interleukin 1, and other proinflammatory cytokines . In vitro experiments with human peripheral blood lymphocytes revealed that pirfenidone reduced SEB-induced cytokine levels 50 to 80% and inhibited 95% of SEB-induced T-cell proliferation . Overall, these studies demonstrated the potential utility of pirfenidone as a therapeutic against septic shock and the biological effects of SEB. Immunol Lett, 2002 Jun 3, 82(1-2), 75 - 8 Superantigen concomitantly induces Th1 cytokine genes and the ability to shut off their expression on re-exposure to superantigen; Arad G et al.; Superantigens, exemplified by staphylococcal enterotoxin B (SEB), are the strongest known inducers of a cellular immune response; they elicit the production of excessive amounts of Th1 cytokines, IL-2, IFN-gamma and TNF, leading to toxic shock . We show that increasing doses of SEB cause not only a greater induction but also a more rapid cessation of IL-2 gene expression . Remarkably, exposure of human PBMC to a second dose of SEB, even at concentrations 10- or 100-fold lower than the initial inducing dose and even within 2 h after the first exposure to SEB, resulted in an immediate and essentially complete shutoff of the induced IL-2 and IFN-gamma mRNA expression . The shutoff response was observed when primary induction of IL-2 and IFN-gamma gene expression was by SEB but not when it was by phytohemaggutinin-P . Signaling by a superantigen thus results not only in a vigorous induction of Th1 cytokine genes but concomitantly induces the ability to shut off their expression upon re-exposure to superantigen . Without induction of this negative control mechanism, the cellular immune response to a superantigen would be even more pronounced. J Vet Med Sci, 2002 Mar, 64(3), 237 - 43 Characterization of Staphylococcus intermedius from pigeons, dogs, foxes, mink, and horses by pulsed-field gel electrophoresis; Wakita Y et al.; Staphylococcus intermedius from pigeons, dogs, foxes, mink, and horses, was characterized by pulsed-field gel electrophoresis (PFGE) to evaluate the use of this typing method for discriminating among strains . SmaI cut the chromosomal DNA into 7-13 fragments ranging from approximately 48 kb to 655 kb, with most of the detectable fragments being smaller than 172 kb . S . intermedius from various animals had a high degree of restriction fragment length polymorphism . Pigeon strains have a similar genotype, despite the difference in their isolation area . Phage typing indicated that the dog, fox, and mink strains belong to the canine I or canine II type . The PFGE method further differentiated the mink strains from the dog and fox strains with regard to three fragments between 256 kb and 570 kb . As such, genomic DNA fingerprinting by PFGE appears to be an effective technique for discriminating S . intermedius strains from various animals . A combination of PFGE typing and phage typing would provide more detailed information than the single method for ecological investigations of S . intermedius. Biosci Biotechnol Biochem, 2002 Feb, 66(2), 328 - 35 Identification of serine138 residue in the 4-residue segment K135K1361137S138 of LukS-I component of Staphylococcus intermedius leukocidin crucial for the LukS-I-specific function of staphylococcal leukocidin; Nishiyama A et al.; Luk-I produced by Staphylococcus intermedius was found to be a new member of the staphylococcal bi-component pore-forming toxin family, in which staphylococcal leukocidin, Panton-Valentine leukocidin, and gamma-hemolysin are included . Luk-I consists of LukS-I and LukF-I . From the deduced amino acid sequence of LukS-I, a 4-residue sequence, K135K1361137S138, at the root of the stem region was found to be identical with that of the phosphorylated segment of a protein phosphorylated by protein kinase A . A mutant of LukS-I (MLSI-SA), in which the Ser138 residue was replaced by an alanine residue, was created, purified, and assayed for its leukocytolytic and pore-forming activities with LukF-I . Both LukS-I and MLSI-SA formed a ring-shaped complex with LukF-I on rabbit erythrocytes and human polymorphonuclear leukocytes (HPMNLs) membrane . However, MLSI-SA showed no leukocytolytic activity with LukF-I . LukS-I was phosphorylated by protein kinase A in the presence of {gamma-32P} ATP in a cell-free system, but MLSI-SA was not phosphorylated significantly . A potent and selective inhibitor of protein kinase A (N-{2(p-bromocinnamylamino)ethyl}-5-isoquinolinesulfonamide (H-89)) showed 50% inhibition of the Luk-I-induced cell lysis at 0.5 nM . Thus, it is concluded that the phosphorylation of the Ser138 residue in the 4-residue segment K135K1361137S138 of LukS-I is important for the leukocytolysis of HPMNLs. Int J Food Microbiol, 2002 May 5, 75(1-2), 61 - 9 Spectral surface plasmon resonance biosensor for detection of staphylococcal enterotoxin B in milk; Homola J et al.; This work evaluates a newly developed wavelength modulation-based SPR biosensor for the detection of staphylococcal enterotoxin B (SEB) in milk . Two modes of operation of the SPR biosensor are described: direct detection of SEB and sandwich assay . In the sandwich assay detection mode, secondary antibodies are bound to the already captured toxin to amplify sensor response . Samples including SEB in buffer and SEB in milk were analyzed in this work . The SPR biosensor has been shown to be capable of directly detecting concentrations of SEB in buffer as low as 5 ng/ml . In sandwich detection mode, the lowest detection limit was determined to be 0.5 ng/ml for both buffer and milk samples . The reported wavelength modulation-based SPR sensor provides a generic platform which can be tailored for detection of various foodborne pathogens and agents for food analysis and testing. Surg Endosc, 2002 May, 16(5), 851 - 4 Epub 2002 Feb 06. Laparoscopic splenectomy; Brodsky JA et al.; BACKGROUND: Laparoscopic splenectomy is currently the procedure of choice for elective splenectomy . This study reviews the initial 100 laparoscopic splenectomies completed at the Cleveland Clinic Foundation . METHODS: A retrospective review of elective laparoscopic splenectomy was performed to assess clinical outcomes at the Cleveland Clinic Foundation . Patient demographics, preoperative diagnoses, operative characteristics, morbidity, and mortality were evaluated . RESULTS: Of the 169 elective splenectomies completed over a 4-year period from 1995 to 1999, 100 were attempted laparoscopically . The proportions of all splenectomies attempted laparoscopically by year were 17%, 38%, 75%, and 72% . Nearly 70% of splenectomies were performed for idiopathic thrombocytopenic purpura or malignancy . Overall, the mean blood loss was 181 ml, and the mean operative time was 170 min . Splenomegaly occurred in 31% of the patients and accounted for longer operative times . Three patients required conversion to an open procedure . Postoperative complications were seen in 13% of the patients . One patient died in the postoperative period from staphylococcal sepsis, giving a mortality rate of 1% . CONCLUSIONS: Laparoscopic splenectomy currently is the procedure of choice for elective splenectomy at our institution . As compared with traditional open splenectomy, laparoscopic splenectomy results in minimal morbidity even in the setting of splenomegaly. J Immunol, 2002 May 15, 168(10), 4832 - 5 Cutting edge: selective impairment of CD8+ T cell function in mice lacking the TNF superfamily member LIGHT; Tamada K et al.; Interactions of LIGHT and its receptors, herpesvirus entry mediator on T cells and lymphotoxin beta receptor on stromal cells, are implicated in the regulation of lymphoid organogenesis, costimulation of T cells, and activation of dendritic cells . In this work we report that LIGHT-deficient mice had normal lymphoid organs with T cells and APCs that normally responded to Ag stimulation and normally stimulated T cells . Although the number of Vbeta8(+) T cells in naive LIGHT(+/+) and LIGHT(-/-) mice was identical, Vbeta8(+)CD8(+) T cell proliferation in response to staphylococcal enterotoxin B was significantly lower in LIGHT(-/-) mice . Consistently, induction and cytokine secretion of CD8(+) CTL to MHC class I-restricted peptide was also reduced in LIGHT(-/-) mice . However, the proliferative response of Vbeta8(+)CD4(+) T cells to staphylococcal enterotoxin B was comparable in LIGHT(-/-) and LIGHT(+/+) mice . Our results suggest that LIGHT is required for activation of normal CD8(+) T cells but not CD4(+) T cells. Clin Exp Allergy, 2002 May, 32(5), 796 - 802 Increased Th1 and Th2 allergen-induced cytokine responses in children with atopic disease; Smart JM et al.; BACKGROUND: Polyclonal cytokine responses following stimulation of T cells with mitogens or superantigens provides information on cytokine production from a wide range of T cells . Alternatively allergen-induced T cell responses can provide information on cytokine production by allergen-reactive T cells . While there is evidence of increased Th2 and reduced Th1 cytokine production following T cell stimulation with non-specific mitogens and superantigens, the evidence that Th1 cytokine production to allergens is decreased in line with a postulated imbalance in Th1/Th2 responses is unclear, with studies finding decreased, no difference or increased IFN-gamma responses to allergens in atopic subjects . OBJECTIVE: To examine childhood polyclonal and allergen-induced cytokine responses in parallel to evaluate cytokine imbalances in childhood atopic disease . METHODS: PBMC cytokine responses were examined in response to a polyclonal stimulus, staphylococcal superantigen (SEB), in parallel with two inhalant allergens, house dust mite (HDM) and rye grass pollen (RYE), and an ingested allergen, ovalbumin (OVA), in (a) 35 healthy children (non-atopic) and (b) 36 children with atopic disease (asthma, eczema and/or rhinitis) (atopic) . RESULTS: Atopic children had significantly reduced IFN-gamma and increased IL-4 and IL-5 but not IL13 production to SEB superantigen stimulation when compared with non-atopic children . HDM and RYE allergens stimulated significantly increased IFN-gamma, IL-5 and IL-13, while OVA stimulated significantly increased IFN-gamma production in atopic children . CONCLUSION: We show that a polyclonal stimulus induces a reduced Th1 (IFN-gamma) and increased Th2 (IL-4 and IL-5) cytokine pattern . In contrast, the allergen-induced cytokine responses in atopic children were associated with both increased Th1 (INF-gamma) and Th2 (IL-5 and IL-13) cytokine production . The increased Th1 response to allergen is likely to reflect prior sensitization and indicates that increases in both Th1 and Th2 cytokine production to allergens exists concomitantly with a decreased Th1 response to a polyclonal stimulus in atopic children. Eur J Vasc Endovasc Surg, 2002 Apr, 23(4), 303 - 8 Endovascular approach to abdominal aortic aneurysms limits the postoperative systemic immune response; Sweeney KJ et al.; OBJECTIVES: endovascular repair of abdominal aortic aneurysms (E-AAA) has in recent years developed as an alternative to the conventional open repair (C-AAA) . Adverse outcomes following the open approach may relate to immune cell activation and the systemic inflammatory response syndrome (SIRS) and organ failure but the benefits in this respect of the endovascular approach are unclear . This study evaluated this question and focused on T-cell activation and function . DESIGN: prospective clinical study . MATERIALS: twenty patients undergoing abdominal aortic aneurysm repair (12 C-AAA and 8 E-AAA) . METHODS: peripheral T-cell expression of surface markers CD69, CD62L and CD25 in vivo and Interleukin 2 (IL-2) and Interleukin-10 (IL-10) responses to the superantigen staphylococcal enterotoxin B (SEB) in vitro were measured preoperatively, 24 h and 1 week postoperatively . RESULTS: there was no significant increase (p=0.23) in the incidence of SIRS in the open compared with the endovascular group . Enhanced T cell activation occurred following C-AAA and this was associated with significantly greater IL-2 production in response to SEB, with no change in IL-10 production . CONCLUSIONS: E-AAA attenuates proinflammatory T-cell changes compared with C-AAA repair . A reduction in T-cell activation and impaired responsiveness to superantigen suggests that the immunological sequelae of the endovascular approach to aneurysm repair is more favourable than after the open approach with potentially less risk of adverse outcomes . Proof of this thesis will require a larger prospective study . Vestn Ross Akad Med Nauk, 2002, (3), 16 - 20 {The role of neutrophils in the regulation of antimicrobial resistance}; Dolgushin II; The functional activity of neutrophils and macrophages and an immune response to sheep red blood cells decrease in mice with local staphylococcal infection in the early period . In the first 24 hours after contamination, the animals' neutrophils lose their capacity to produce immuno-stimulating agents, they begin secreting immunosuppressive factors . Administration of secretory products of activated neutrophils, the low-weight molecular peptide isolated from these cells to the induced animals restores the immunological responsiveness of the staphylococcus-infected animals, reduces the death of mice infected with S . aureus and S . typhimurium. Neurosci Lett, 2002 May 17, 324(2), 93 - 6 Focal encephalitis in the Lewis rat induced by intracerebral enterotoxin superantigen and amplified by activated intravenous splenocytes; Kornhuber ME et al.; The histological response after intracerebral (IC) injection of superantigen (SAg) was investigated in unprimed Lewis rats . The staphylococcal enterotoxins (SE) A (SEA), and E, but not B or saline, induced a variable perivascular inflammation in the injected hemisphere (6.7 +/- 6.1 cuffs per tissue section with SEA; mean +/- SD) . Adoptive transfer of mitogen activated splenocytes (AS) augmented the response to SEA significantly (18.5 +/- 11.4; P<0.05) . With or without AS transient bilateral perivascular cuffs were observed around the ventricles and in the corpus callosum up to 3 days after IC injection . The findings demonstrate that local expression of SAg in the brain can cause encephalitis, depending on the number of activated T cells in the circulation. Graefes Arch Clin Exp Ophthalmol, 2002 Apr, 240(4), 265 - 70 Epub 2002 Mar 02. Internal and external contamination of donor corneas before in situ excision: bacterial risk factors in 93 donors; Robert PY et al.; BACKGROUND: Most studies of corneal donor contamination concentrate on postenucleation contamination of the eyeball . The purpose of the present study was to evaluate the relative contamination of in situ excised corneal tissue and relevance to final success or rejection by recipients of the corneal grafts . METHODS: Ninety-three donors underwent anterior chamber puncture (ACP) and corneal epithelium scarification (CS) before and after disinfection with 5% povidone-iodine . Following in situ excision, corneas were preserved in culture medium at +35 degrees C . Morphological and bacteriological assessment was carried out after culture, and recipients were followed up over a 2-year period . RESULTS: Samples taken by ACP, CS before disinfection, CS after disinfection and a culture medium sample were contaminated by bacteria in, respectively, 8 (8.6%), 23 (24.7%) 4 (4.3%) and 5 (5.4%) donors . Contamination of aqueous humor was significantly associated with age, death-to-sample time and premortem systemic infection . Contamination of epithelium significantly increased culture medium contamination . CONCLUSION: External bacteria on donor cornea are mainly skin bacteria (especially Staphylococcus) and can be partially eliminated by a povidone-iodine wash . Internal bacteria are mainly gut bacteria and may be due to perimortem bacteriemia . However, bacterial infection at the time of death appears to have no effect on the incidence of endophthalmitis in recipients and should no longer prevent use of such corneal tissue in grafts. J Clin Microbiol, 2002 May, 40(5), 1824 - 5 Molecular identification of Staphylococcus lugdunensis in a patient with meningitis; Kaabia N et al.; A 12-year-old child developed meningitis 6 days after a third ventriculostomy by endoscopy . A coagulase-negative Staphylococcus sp . was isolated in pure culture from the cerebrospinal fluid and was definitely identified as Staphylococcus lugdunensis after the 16S ribosomal DNA gene and rpoB gene were sequenced . This report describes the first case of S . lugdunensis meningitis. Klin Med (Mosk), 2002, 80(3), 28 - 33 {Combination of Libman-Sacks endocarditis with infectious endocarditis}; Demin AA et al.; Out of 242 patients treated for systemic lupus erythematosus (SLE) in Novosibirsk for 15 years, valvular lesions and endocarditis were diagnosed in 41(16.9%) patients . Combination of Libman-Sax endocarditis (LSE) with infectious endocarditis (IE) was observed in three patients (two women, one man, age 18-40 year) . SLE ran a subacute course in one woman, an acute one--in the other . LSE emerged early in SLE in two patients . All the patients had polyorganic lupus pathology, lupus nephritis with nephrotic syndrome (morphological class IV) . Two patients had mitral valve disease, one patient--mitral-aortic disease . The rise of secondary IE was seen after massive immunosuppressive therapy . The diagnosis of secondary IE was made after SLE duration for 10-36 months . At IE diagnosis, all the patients had high titers of blood antiphospholipid antibodies . IE was of staphylococcal origin in two patients and candidosis-induced in one patient . In SLE with IE there was thromboembolic syndrome . LSE and IE have related aspects which should be regarded in clinical practice: possible "IE mask" in LSE, risk of secondary IE in about 10% of LSE patients, prophylactic measures necessary to prevent IE in hemodynamically prominent forms of LSE. J Cataract Refract Surg, 2002 May, 28(5), 898 - 9 Delayed-onset multifocal polymicrobial keratitis after laser in situ keratomileusis; Ritterband D et al.; We report a case of mixed Aspergillus fumigatus and coagulase-negative Staphylococcus stromal keratitis in a 43-year-old man who developed discomfort and swelling in his right eye 20 days after uneventful bilateral laser in situ keratomileusis (LASIK) . Clinical examination revealed 2 distinct corneal infiltrates beneath the LASIK flap . Corneal scrapings were taken for microscopic examination and culture . Both infiltrates initially improved on topical antibiotic therapy, but over the next 18 days, 1 infiltrate worsened and repeat cultures were performed, which demonstrated A fumigatus . Natamycin 5% and amphotericin 0.1% were started and continued for 8 weeks with resolution of the infiltrate and return of the best corrected visual acuity . Delayed-onset multifocal keratitis is a rare complication of LASIK . Fungal entities should be considered in the differential diagnosis. Ann Dermatol Venereol, 2002 Jan, 129(1 Pt 2), 147 - 51 {Cutaneous hydration and atopia}; Stalder JF; Altered barrier function of atopic skin is characterized by xerosis, which predominantly affects non-inflammatory areas . Xerosis is accompanied by morphological, physical and biochemical modifications . The composition of epidermal lipids in atopic patients is modified both qualitatively and quantitatively . The impact of xerosis and abnormal barrier function in such patients is on different levels: --via excessive permeability to allergens and atopens, --but also via constant staphylococcal colonization of the atopic skin . The role of moisturizing treatments in restoring the barrier function in such patients is an important therapeutic aim . Clinical assessment of the effect of moisturizing topicals relies on objective means (SCORAD index) but also on avoiding corticosteroids or the impact of these moisturizers on patients'quality of life. J Neurol Neurosurg Psychiatry, 2002 May, 72(5), 644 - 6 Immunoadsorption inferior to plasma exchange in a patient with chronic inflammatory demyelinating polyradiculoneuropathy; Hadden RD et al.; Staphylococcal protein A immunoadsorption and plasma exchange were compared for treating chronic inflammatory demyelinating polyradiculoneuropathy . In a single patient, plasma exchange had a more beneficial effect than immunoadsorption on clinical outcome measures . Serum IgM antibody activity to peripheral nerve fell significantly following plasma exchange . Serum IgM and IgA fell more and IgG less after plasma exchange than after immunoadsorption . The superior efficacy of plasma exchange to immunoadsorption in this case may have been the result of removal of an IgM antibody. J Immunol, 2002 May 1, 168(9), 4352 - 60 T cell-dependent maturation of dendritic cells in response to bacterial superantigens; Muraille E et al.; Dendritic cells (DC) express a set of germline-encoded transmembrane Toll-like receptors that recognize shared microbial products, such as Escherichia coli LPS, termed pathogen-associated molecular patterns . Analysis of the in vivo response to pathogen-associated molecular patterns has uncovered their ability to induce the migration and the maturation of DC, favoring thus the delivery of Ag and costimulatory signals to naive T cells in vivo . Bacterial superantigens constitute a particular class of pathogen-derived molecules known to induce a potent inflammatory response in vivo, secondary to the activation of a large repertoire of T cells . We demonstrate in this work that Staphylococcal superantigens induce migration and maturation of DC populations in vivo . However, in contrast to LPS, superantigens failed to induce DC maturation in RAG or MHC class II-deficient mice, suggesting that T cell activation was a prerequisite for DC maturation . This conclusion was further supported by the finding that T cell activation induced by 1) mitogenic anti-CD3 mAbs, 2) allo-MHC determinants, or 3) nominal Ag in a TCR-transgenic model induces DC maturation in vivo . These studies also revealed that DC that matured in response to T cell mitogens display, comparatively to LPS, a distinctive phenotype characterized by high expression of the MHC class II, CD40, and CD205 markers, but only moderate (CD86) to minimal (CD80) expression of CD28/CTLA4 ligands . This work demonstrates that activation of a sufficient number of naive T cells in vivo constitutes a novel form of immune danger, functionally linked to DC maturation. Biochemistry, 2002 Apr 30, 41(17), 5656 - 67 Electrostatic effects in highly charged proteins: salt sensitivity of pKa values of histidines in staphylococcal nuclease; Lee KK et al.; The pK(a) values of most histidines in small peptides and in myoglobin increase on average by 0.30 unit between 0.02 and 1.5 M NaCl {Kao et al . (2000) Biophys . J . 79, 1637} . The DeltapK(a) values reflect primarily the ionic strength dependence of the solvation energy; screening of Coulombic interactions contributes only in a minor way . This implies that Coulombic interactions are weak, or that attractive and repulsive contributions to the pK(a) values are balanced . To distinguish experimentally between these two possibilities, and to further characterize the magnitude and salt sensitivity of surface electrostatic interactions in proteins, the salt dependence of pK(a) values of histidines in staphylococcal nuclease was measured by (1)H NMR spectroscopy . Three of the four histidines titrated with significantly depressed pK(a) values, and the salt sensitivity of all histidine pK(a) values was substantial . In three cases, the pK(a) values increased by a full unit between 0.01 and 1.5 M KCl . Anion-specific effects were found; the pK(a) values measured under equivalent ionic strengths in SCN(-) and SO(4)(2-) were higher than in Cl(-); the order of the sensitivity of pK(a) values to anions was SCN(-) > Cl(-) > SO(4)(2-) . Structure-based pK(a) calculations with continuum methods were performed to interpret the measured effects structurally and to test their ability to capture the experimental behavior . Calculations in which the protein interior was treated empirically with a dielectric constant of 20 reproduced the pK(a) values and their dependence on the concentration of Cl(-) . According to the calculations, the pK(a) values are depressed because of unfavorable self-energies and repulsive Coulombic interactions . Their striking salt sensitivity reflects screening of weak, repulsive, Coulombic interactions among charges separated by more than 10 A . Long-range Coulombic interactions on the surfaces of proteins are weak, but they can add up to produce substantial electrostatic effects when positive and negative charges are not balanced. Issues Emerg Health Technol, 2002 Jan, (28), 1 - 4 Extracorporeal immunoadsorption treatment for rheumatoid arthritis; Hailey D et al.; Immunoadsorption treatment is a non-drug therapy for rheumatoid arthritis . The treatment is based on filtering the patient's plasma through a column containing staphylococcal protein A . The treatment is effective in alleviating the symptoms of severe rheumatoid arthritis in some patients . Data on long-term outcomes are not available . The mechanism of action of this treatment is unclear . Most adverse effects are associated with the apheresis procedure . The cost per 12 week course of treatment is likely to be more than C $20,000 . The cost-effectiveness of the technology is not yet established. Protein Sci, 2002 May, 11(5), 1004 - 16 Distance dependence and salt sensitivity of pairwise, coulombic interactions in a protein; Lee KK et al.; Histidine pK(a) values were measured in charge-reversal (K78E, K97E, K127E, and K97E/K127E) and charge-neutralization (E10A, E101A, and R35A) mutants of staphylococcal nuclease (SNase) by (1)H-NMR spectroscopy . Energies of interaction between pairs of charges (DeltaG(ij)) were obtained from the shifts in pK(a) values relative to wild-type values . The data describe the distance dependence and salt sensitivity of pairwise coulombic interactions . Calculations with a continuum electrostatics method captured the experimental DeltaG(ij) when static structures were used and when the protein interior was treated empirically with a dielectric constant of 20 . The DeltaG(ij) when r(ij) < or = 10 A were exaggerated slightly in the calculations . Coulomb's law with a dielectric constant near 80 and a Debye-Huckel term to account for screening by the ionic strength reproduced the salt sensitivity and distance dependence of DeltaG(ij) as well as the structure-based method . In their interactions with each other, surface charges behave as if immersed in water; the Debye length describes realistically the distance where interactions become negligible at a given ionic strength . On average, charges separated by distances (r(ij)) approximately 5 A interacted with DeltaG(ij) approximately 0.6 kcal/mole in 0.01 M KCl, but DeltaG(ij) decayed to < or =0.10 kcal/mole when r(ij) = 20 A . In 0.10 M KCl, DeltaG(ij) approximately 0.10 kcal/mole when r(ij) = 10 A . In 1.5 M KCl, only short-range interactions with r(ij) < or = 5 A persisted . Although at physiological ionic strengths the interactions between charges separated by more than 10 A are extremely weak, in situations where charge imbalance exists many weak interactions can cumulatively produce substantial effects. J Virol, 2002 May, 76(10), 4741 - 9 Frequencies of virus-specific CD4(+) and CD8(+) T lymphocytes secreting gamma interferon after acute natural rotavirus infection in children and adults; Jaimes MC et al.; Human rotavirus-specific CD4(+) and CD8(+) T-cell responses in peripheral blood lymphocytes were studied using a flow cytometric assay that detects the intracellular accumulation of cytokines after short-term in vitro antigen stimulation . The frequencies of virus-specific T cells that secrete gamma interferon and interleukin-13 (IL-13) were determined in adults and children during the acute or convalescent phase of rotavirus-induced diarrhea, in asymptomatically infected adults and laboratory workers who worked with human stool samples containing rotavirus, and in healthy adults . Significantly higher frequencies of rotavirus-specific interferon gamma-secreting CD8(+) and CD4(+) T cells, but not IL-13-secreting T cells, were detected in symptomatically infected adults and exposed laboratory workers than in healthy adults and children with acute rotavirus diarrhea . The levels of rotavirus-specific T cells returned to levels found in healthy adults by 32 days after the onset of rotavirus diarrhea in most adult subjects . Children with rotavirus diarrhea had undetectable or very low levels of CD4(+) and CD8(+) T cells that secrete gamma interferon . Adult cytomegalovirus-seropositive individuals had frequencies of cytomegalovirus-specific T cells that secrete gamma interferon that were approximately 20 times the level of rotavirus-specific T cells . This result suggests that rotavirus is a relatively poor inducer of circulating memory T cells that secrete gamma interferon . The frequencies of gamma interferon-secreting CD4(+) and CD8(+) T cells and the frequencies of IL-13-secreting CD4(+) T cells responding to the T-cell superantigen staphylococcal enterotoxin B (SEB) were lower in children than in adults . In both adults and children, the frequencies of CD4(+) cells secreting gamma interferon in response to SEB were higher than the frequencies of cells secreting IL-13. Scand J Immunol, 2002 Apr, 55(4), 373 - 81 Costimulatory molecule requirement for bovine WC1+gammadelta T cells' proliferative response to bacterial superantigens; Fikri Y et al.; We have previously shown that the proliferation of freshly isolated bovine WC1+gammadelta T cells to superantigens (SAgs) including staphylococcal enterotoxin A (SEA), and staphylococcal enterotoxin B (SEB) or toxic shock syndrome type-1 (TSST-1) required the presence of antigen-presenting cells (APC) and the addition of exogenous interleukin (IL)-2 . The costimulatory activity provided by molecules expressed on professional APC for the proliferation of gammadelta T cells has not been addressed hitherto . In the present study, we investigated the ability of two selected APC populations, the dendritic cells (DCs) highly expressing CD80 and CD86 molecules (CD80highCD86high) and the monocytes expressing the same molecules at a rather low level (CD80lowCD86low), to stimulate the proliferation of purified bovine WC1+gammadelta T cells to SAgs . DCs were more efficient than monocytes in inducing gammadelta T-cell proliferation, and this response was dependent on exogenous IL-2 in both presentation modes . Stimulating gammadelta T cells with gradual doses of SAgs or concanavalin A (ConA) resulted in similar dose-dependent reaction profiles suggesting a minimal role of the major histocompatibility complex (MHC) . However, significant proliferation was already obtained with the starting doses in the presence of DC compared with monocytes, and higher proliferation was reached with DC at optimal doses . Finally, the addition of monoclonal antibody (MoAb) anti-CD86 markedly inhibited SAgs- and ConA-mediated proliferation, whereas MoAb anti-CD80 had no effect . The combination of both anti-CD80 and anti-CD86, however, suppressed this response . These results suggest that bovine gammadelta T-cell proliferation response requires indubitably CD86 costimulation . The role of CD80 molecule seems less clear. Biosens Bioelectron, 2002 Jun, 17(6-7), 605 - 18 An integrated, stacked microlaboratory for biological agent detection with DNA and immunoassays; Yang JM et al.; An integrated, stacked microlaboratory for performing automated electric-field-driven immunoassays and DNA hybridization assays was developed . The stacked microlaboratory was fabricated by orderly laminating several different functional layers (all 76 x 76 mm(2)) including a patterned polyimide layer with a flip-chip bonded CMOS chip, a pressure sensitive acrylic adhesive (PSA) layer with a fluidic cutout, an optically transparent polymethyl methacrylate (PMMA) film, a PSA layer with a via, a patterned polyimide layer with a flip-chip bonded silicon chip, a PSA layer with a fluidic cutout, and a glass cover plate layer . Versatility of the stacked microlaboratory was demonstrated by various automated assays . Escherichia coli bacteria and Alexa-labeled protein toxin staphylococcal enterotoxin B (SEB) were detected by electric-field-driven immunoassays on a single chip with a specific-to-nonspecific signal ratios of 4.2:1 and 3.0:1, respectively . Furthermore, by integrating the microlaboratory with a module for strand displacement amplification (SDA), the identification of the Shiga-like toxin gene (SLT1) from E . coli was accomplished within 2.5 h starting from a dielectrophoretic concentration of intact E . coli bacteria and finishing with an electric-field-driven DNA hybridization assay, detected by fluorescently labeled DNA reporter probes . The integrated microlaboratory can be potentially used in a wide range of applications including detection of bacteria and biowarfare agents, and genetic identification. Biosens Bioelectron, 2002 Jun, 17(6-7), 591 - 5 A miniature fiber optic surface plasmon resonance sensor for fast detection of Staphylococcal enterotoxin B; Slavik R et al.; A fiber optic surface plasmon resonance (SPR) biosensor for detection of Staphylococcal enterotoxin B (SEB) is reported . The sensor is based on spectral interrogation of surface plasmons in a miniature sensing element based on a side-polished single-mode optical fiber with a thin metal overlayer . For specific detection of SEB, the SPR sensor is functionalized with a covalently crosslinked double-layer of antibodies against SEB . The SPR biosensor is demonstrated to be able to detect ng/ml concentrations of SEB in less than 10 min. J Agric Food Chem, 2002 Apr 24, 50(9), 2471 - 4 Development of an enzymatic method to quantify methyl ketones from bacterial origin; Fadda SG et al.; Methyl ketones are detected in dry fermented sausages in which they contribute to the cured aroma . They have been associated with the inoculation of Staphylococcus carnosus used as starter culture.To evaluate the ability of bacterial starters to produce methyl ketones it was necessary to develop a rapid method . The method consists of a reaction catalyzed by a commercial NADPH-dependent alcohol dehydrogenase that reduces the 2-pentanone to its secondary alcohol . The linearity, the specificity, and the robustness were studied . Its accuracy was confirmed by comparison with the gas chromatography technique . Finally, the method was validated on biological samples such as the 2-pentanone produced by Staphylococcus carnosus . The enzymatic method offers some advantages over the gas chromatography, as it is faster, simpler, and inexpensive, guaranteeing an effective way to assess bacterial ketone production. J Cataract Refract Surg, 2002 Apr, 28(4), 703 - 8 Effect of vancomycin on Staphylococcus epidermidis adherence to poly(methyl methacrylate) intraocular lenses; Das T et al.; PURPOSE: To study the adherence of Staphylococcus epidermidis in variable loads (10(8) cfu/mL and 10(3) cfu/mL) to poly(methyl methacrylate) (PMMA) intraocular lenses (IOLs) and to evaluate the effect of vancomycin treatment on S epidermidis adherence to the IOL . SETTING: L.V . Prasad Eye Institute, Hyderabad, India . METHODS: The study was designed in 2 parts . Phase I: Twelve PMMA IOLs were soaked in a solution of S epidermidis, randomized to 10(8) cfu/mL or 10(3) cfu/mL . They were rinsed or rinsed and vortexed and cultured . Phase II: Twelve IOLs were treated with vancomycin (10 mg/mL), randomized to before and after treatment with S epidermidis . RESULTS: Staphylococcus epidermidis adhered to all portions of the IOL with 10(8) cfu/mL and 10(3) cfu/mL bacterial loads . Treatment with vancomycin reduced S epidermidis adherence . CONCLUSION: Pretreatment of an IOL with vancomycin or a suitable antibiotic agent appears to reduce bacterial adherence to the IOL . Placement of a sustained-release device that could release an antibiotic agent over a specific period, reducing the incidence of postcataract endophthalmitis, should be studied. Vet Microbiol, 2002 May 24, 86(4), 281 - 93 Exudative epidermitis and porcine circovirus-2 infection in a Swedish SPF-herd; Wattrang E et al.; An outbreak of exudative epidermitis (EE) among piglets in a Swedish SPF-herd initiated a survey for indications as to the cause of disease.The herd was established by caesarean section and has been closed to all new animal material, with the exception of semen for artificial insemination (AI) . The study comprised serum samples from the SPF-herd over a 10-year period (n=109) and a close monitoring of animals in the herd during the period after the EE outbreak . Serum samples from conventional boars at the AI-station servicing the herd were also included (n=9) . All serum samples were tested for antibodies to porcine circovirus-2 (PCV-2) . In addition, 3-week-old piglets from three litters (n=24) farrowed close after the initial EE outbreak were closely monitored for clinical signs of skin disease, sampled for Staphylococcus hyicus, tested for antibodies to porcine parvovirus and in sequentially collected serum samples tested for interferon-alpha (IFN-alpha) and interleukin-6.The PVC-2 serology showed that animals in the herd were sero-negative at least until 2 months prior to the EE outbreak . During the period close after the EE outbreak the animals showed varying levels of antibodies to PCV-2 but all the tested animals had sero-converted 4 months later . The AI boars were also sero-positive to PCV-2 at the time of the EE outbreak . Animals in the SPF-herd remained sero-positive to PCV-2 during the following 7 years . In the monitored litters, one piglet had clinical EE and 15 piglets displayed defined erythemas on the abdomen . Fourteen of the piglets also had IFN-alpha in serum on one or more occasions during the study, indicating viral activity among the animals . S . hyicus was isolated from all of the piglets from the earliest sampling point (3 days of age) and onwards, irrespective of clinical signs . PCV-2 was isolated from lymph node tissue collected from one of the EE affected pigs.Further, increases in the number of stillborn piglets, small litters (<6 piglets) and repeat breeders could be correlated to the time of PCV-2 sero-conversion . Coincidence of active viral infection and sero-conversion to PCV-2 points to the virus as the cause of the EE outbreak and reproductive disturbances. Arthritis Rheum, 2002 Apr, 46(4), 1028 - 38 Defective Fc-dependent processing of immune complexes in patients with systemic lupus erythematosus; Davies KA et al.; OBJECTIVE: To explore the Fc receptor-dependent handling of immune complexes (ICs) by the fixed mononuclear phagocytic systems (MPS) in patients with systemic lupus erythematosus (SLE) . METHODS: The processing in vivo of soluble model (123)I-hepatitis B/ anti-hepatitis B ICs was studied in 12 healthy subjects and 10 patients with active SLE . ICs that fixed complement poorly were prepared specifically in order to explore Fc receptor-dependent clearance mechanisms . Clearance kinetics and organ uptake were assessed by computer-aided gamma scintigraphy and serial blood sampling . RESULTS: In both patients and controls, the main site of IC clearance was the liver; only 2-6% of injected ICs were taken up in the spleen . The kinetics of initial IC clearance were similar in both groups, but defective hepatic retention of ICs was demonstrated in patients with SLE . At 1 hour, hepatic activity in patients had fallen to 56% of maximum, compared with 74% in controls (P = 0.0002) . Precipitation studies performed on serum samples using staphylococcal protein A-Sepharose indicated that antibody-complexed tracer was released from the liver 20-50 minutes after injection . CONCLUSION: These results indicate that Fc-mediated clearance of ICs is defective in patients with SLE and suggest that ligation of ICs by Fc receptors is critical for their efficient binding and retention by the fixed MPS in the liver. Infect Immun, 2002 May, 70(5), 2278 - 81 Protection against bacterial superantigen staphylococcal enterotoxin B by passive vaccination; LeClaire RD et al.; We investigated the ability of two overlapping fragments of staphylococcal enterotoxin B (SEB), which encompass the whole toxin, to induce protection and also examined if passive transfer of chicken anti-SEB antibodies raised against the holotoxin could protect rhesus monkeys against aerosolized SEB . Although both fragments of SEB were highly immunogenic, the fragments failed to protect mice whether they were injected separately or injected together . Passive transfer of antibody generated in chickens (immunoglobulin Y {IgY}) against the whole toxin suppressed cytokine responses and was protective in mice . All rhesus monkeys treated with the IgY specific for SEB up to 4 h after challenge survived lethal SEB aerosol exposure . These findings suggest that large fragments of SEB may not be ideal for productive vaccination, but passive transfer of SEB-specific antibodies protects nonhuman primates against lethal aerosol challenge . Thus, antibodies raised in chickens against the holotoxin may have potential therapeutic value within a therapeutic window of opportunity after SEB encounter. J Arthroplasty, 2002 Apr, 17(3), 355 - 8 The clinical significance of femoral head culture results in donors after hip arthroplasty: a preliminary report; James LA et al.; The aim of this study is to determine whether a link exists between clinical wound problems and positive bacterial culture of the femoral head after primary hip arthroplasty . Clinical data were retrieved for 24 culture-positive and 26 culture-negative patients . Coagulase-negative Staphylococcus was the commonest pathogen, cultured in 19 of the 24 culture-positive femoral heads donated to a regional bone bank . Wound problems occurred in 2 of the patients from the culture-positive group and 4 of the patients from the culture-negative group . The relative risk of wound problems was 0.54 in the culture-positive group . Based on the available evidence in this preliminary short-term retrospective review, positive microbiology from donated femoral heads has no clinical implication in the donors . The practice of reporting culture-positive femoral heads should be continued. Saudi Med J, 2002 Feb, 23(2), 229 - 31 Staphylococcal toxic shock syndrome in a small infant; Ghazal SS et al.; Non menstrual staphylococcal toxic shock syndrome is rare in small infants . This is a 4-month-old infant presented to us with a picture of bronchiolitis and few postuler skin lesions, treated with antistapylococcal antibiotics in addition to other supportive medications . On the 4th day of therapy the patient developed sunburn like erythroderma, hypotension, and high grade fever . The dose of antibiotics was increased to the maximum possible dose, in addition to other supportive medications . The patient improved and developed extensive desquamation in both hands and feet on the 14th day of hospitalization, which confirms the diagnosis. Eur Respir J, 2002 Mar, 19(3), 447 - 57 Effects of inducible nitric oxide synthase and xanthine oxidase inhibitors on SEB-induced interstitial pneumonia in mice; Miyakawa H et al.; The authors have previously reported that intratracheal instillation of staphylococcal enterotoxin-B (SEB) induced interstitial pneumonia (IP) in autoimmune-prone mice . SEB-reactive T-cells were critically involved in the development of IP in this model . Concern has arisen about the hazards of reactive oxygen species (ROS) and reactive nitrogen species (RNS) in the process of lung injury and fibrosis . Therefore, the involvement of nitric oxide (NO) and superoxide anion (O2-) in the pathogenesis of IP in this autoimmune-prone model has been investigated . Nitrite/nitrate levels were increased in bronchoalveolar lavage (BAL) fluid and serum from SEB-injected mice . The signal of the NO-(N-(dithiocarboxy) sarcosine)2-Fe2+ complex was detected in the SEB-injected lung and whole blood by electron paramagnetic resonance (EPR) spectroscopy . NO production was significantly decreased by aminoguanidine (AG) treatment . Xanthine oxidase (XO) activity in the lung, BAL fluid, and plasma was increased with instillation of SEB, and 4-amino-6-hydroxypyrazolo(3,4-d)-pyrimidine (AHPP) significantly inhibited XO activity . Moreover, both AG and AHPP significantly decreased production of pro-inflammatory cytokines, numbers of infiltrated cells in BAL fluid, and the area of thickened alveolar septa in the SEB-injected lung . In conclusion, the overproduction of nitric oxide and super oxide anion were implicated in the pathogenesis of interstitial pneumonia, and inducible nitric oxide synthase and xanthine oxidase inhibitors had protective effects against interstitial pneumonia in this model. Graefes Arch Clin Exp Ophthalmol, 2002 Mar, 240(3), 220 - 6 Epub 2002 Feb 16. Morphological evidence of M cells in healthy canine conjunctiva-associated lymphoid tissue; Giuliano EA et al.; PURPOSE: To characterize the follicle-associated epithelium (FAE) and organized lymphoid nodules from dog nictitating membranes to determine if canine conjunctiva-associated lymphoid tissue (CALT) contains M cells analogous to those described in other regions of mucosa-associated lymphoid tissue (MALT) . METHODS: Nictitan lymphoid follicles from 15 healthy dogs (30 eyes) were harvested immediately post-mortem . Twelve follicles from each nictitating membrane were isolated . Four follicles from each eye of 10 dogs were examined by light microscopy, transmission electron microscopy and scanning electron microscopy . Five of the 10 dogs were treated with a heat-killed staphylococcal topical suspension immediately prior to euthanasia . Nictitan follicles from five other dogs were processed for immunohistochemistry to characterize follicle lymphocyte populations . RESULTS: The FAE overlying CALT demonstrated morphology characteristic of M cells, including attenuated apical cell surface with blunted microvilli and microfolds, invaginated basolateral membrane forming a cytoplasmic pocket containing lymphocytes and macrophages, and diminished distance between the apical and pocket membrane . Heat-killed bacteria were bound to the surface and transcytosed to the cytoplasmic pocket of CALT M cells . Immunohistochemistry of organized lymphoid tissue subtending the FAE demonstrated B-cell germinal centers with T-cell predominant apical caps . CONCLUSIONS: In canine CALT, the FAE overlying lymphoid follicles, as well as the distribution of T and B lymphocytes subtending this region, contain morphologic and functional features analogous to MALT described in other regions . Documentation of canine conjunctival M cells is of clinical relevance in the study of primary ocular diseases, as well as a potential means of vaccination or drug delivery. J Biol Chem, 2002 Jun 21, 277(25), 22839 - 46 Epub 2002 Apr 04. Zinc-mediated dimerization and its effect on activity and conformation of staphylococcal enterotoxin type C; Chi YI et al.; Staphylococcal enterotoxins are superantigen exotoxins that mediate food poisoning and toxic shock syndrome in humans . Despite their structural and functional similarities, superantigens display subtle differences in biological properties and modes of receptor binding as a result of zinc atoms bound differently in their crystal structures . For example, the crystal structures of the staphylococcal enterotoxins in the type C serogroup (SECs) contain a zinc atom coordinated by one aspartate and two histidine residues from one molecule and another aspartate residue from the next molecule, thus forming a dimer . This type of zinc ligation and zinc-mediated dimerization occurs in several SECs, but not in most other staphylococcal enterotoxin serogroups . This prompted us to investigate the potential importance of zinc in SEC-mediated pathogenesis . Site-direct