Staphylococcus

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-directed mutagenesis was used to replace SEC zinc binding ligands with alanine . SEC mutants unable to bind zinc did not have major conformational alterations although they failed to form dimers . Zinc binding was not essential for T cell stimulation, emesis, or lethality although in general the mutants were less pyrogenic . Thus the zinc atom in SECs might represent a non-functional heavy atom in an exotoxin group that has diverged from related bacterial toxins containing crucial zinc atoms.

Proteins, 2002 May 1, 47(2), 99 - 105
Folding of a small helical protein using hydrogen bonds and hydrophobicity forces; Favrin G et al.; A reduced protein model with five to six atoms per amino acid and five amino acid types is developed and tested on a three-helix-bundle protein, a 46-amino acid fragment from staphylococcal protein A . The model does not rely on the widely used Go approximation, which ignores non-native interactions . We find that the collapse transition is considerably more abrupt for the protein A sequence than for random sequences with the same composition . The chain collapse is found to be at least as fast as helix formation . Energy minimization restricted to the thermodynamically favored topology gives a structure that has a root-mean-square deviation of 1.8 A from the native structure . The sequence-dependent part of our potential is pairwise additive . Our calculations suggest that fine-tuning this potential by parameter optimization is of limited use .

Eur J Cardiothorac Surg, 2002 Apr, 21(4), 644 - 8
Prevention of recurrent empyema after pneumonectomy for chronic infection; Schneiter D et al.; OBJECTIVES: Pneumonectomy in chronic pulmonary infection with empyema is associated with a high mortality rate and an increased risk of recurrent empyema . The surgical resection is technically demanding, and successful management continues to be a challenge . METHODS: We evaluated a concept which combines (pleuro-)pneumonectomy or completion pneumonectomy with surgical debridement of the pleural cavity and packing with povidine-iodine soaked dressings . The debridement and packing is repeated in the operating theater after 48 h until the chest cavity is macroscopically clean . Finally, the pleural space is obliterated with antibiotic solution . RESULTS: Between February 1997 and October 2000, 11 patients (average age of 59 years, ranging from 25 to 84) with destroyed lung caused by tuberculosis (six), aspergilloma (two), bronchiectasis (one), esophago-pleural fistula (one) or broncho-pleural fistula after lobectomy for bronchial carcinoma (one) and ongoing chronic infection with acute empyema (ten) (25-2500 days between first and definitive therapy) were treated . Pleural culture findings showed Aspergillus in four, Mycobacterium in two, Enterococcus in two, Candida in one and Staphylococcus in one, respectively . The mean number of interventions was 2.9 (2-4) . The chest was definitively closed in all patients within 1 week . The mean hospitalization time was 19 days (9-31 days).In the follow-up (10-54 months), there was no recurrence of empyema . One patient (84 years) died at day 31, due to sepsis . CONCLUSIONS: Pneumonectomy combined with repeated surgical debridement and antimicrobial therapy enables the successful treatment of chronic pulmonary infection with empyema within a short time period.

J Mol Microbiol Biotechnol, 2002 May, 4(3), 309 - 14
Carbon catabolite repression by the catabolite control protein CcpA in Staphylococcus xylosus; Jankovic I et al.; Carbon catabolic repression (CR) by the catabolite control protein CcpA has been analyzed in Staphylococcus xylosus . Genes encoding components needed to utilize lactose, sucrose, and maltose were found to be repressed by CcpA . In addition, the ccpA gene is under negative autogenous control . Among several tested sugars, glucose caused strongest CcpA-dependent repression . Glucose can enter S . xylosus in nonphosphorylated form via the glucose uptake protein GlcU . Internal glucose is then phosphorylated by the glucose kinase GlkA . Alternatively, glucose can be transported and concomitantly phosphorylated by glucose-specific permease(s) of the phosphotransferase system (PTS) . S . xylosus mutant strains deficient in GlcU or GlkA showed partial relief of glucose-specific, CcpA-dependent repression . Likewise, blocking PTS activity completely by inactivation of the gene encoding the general PTS protein enzyme I resulted in diminished glucose-mediated repression . Thus, both glucose entry routes contribute to glucose-specific CR in S . xylosus . The sugar transport activity of the PTS is not required to trigger glucose-specific repression . The phosphocarrier protein HPr however, is absolutely essential for CcpA activity . Inactivation of the HPr gene led to a complete loss of CR . Repression is also abolished upon inactivation of the HPr kinase gene or by replacing serine at position 46 of HPr by alanine . These results clearly show that HPr kinase provides the signal, seryl-phosphorylated HPr, to activate CcpA in S . xylosus.

J Infect Dis, 2002 Apr 15, 185(8), 1192 - 6 Epub 2002 Apr 01.
Immune protection against staphylococcal enterotoxin-induced toxic shock by vaccination with a Venezuelan equine encephalitis virus replicon; Lee JS et al.; A candidate vaccine against staphylococcal enterotoxin B (SEB) was developed using a Venezuelan equine encephalitis (VEE) virus vector . This vaccine is composed of a self-replicating RNA, termed "replicon," containing the VEE nonstructural genes and cis-acting elements and a gene encoding mutagenized SEB (mSEB) . Cotransfection of baby hamster kidney cells with the mSEB replicon and 2 helper RNA molecules resulted in the release of propagation-deficient mSEB-VEE replicon particles (mSEB-VRPs) . Mice inoculated subcutaneously with mSEB-VRPs were protected (15 of 20 mice) from a challenge with 5 median lethal dose units of wild-type (wt) SEB . T cells from mice vaccinated with mSEB-VRP responded normally both in vitro to wt SEB and in recall response to the inactivated mSEB polypeptide . The profile of cytokines measured after challenge with wt SEB suggested that the mode of protection was predominantly Th1 dependent . Our results suggest that the VEE replicon is a practical and convenient model system for evaluating efficacy of vaccines for the control of bacterial diseases.

Planta, 2002 Feb, 214(4), 546 - 51
Characterization and modelling of the hydrophobic domain of a sunflower oleosin; Alexander LG et al.; The oleosins are a group of hydrophobic proteins present on the surface of oil bodies in seeds, where they are thought to prevent coalescence . They contain a central hydrophobic domain of 68-74 residues that is thought to form a loop into the triacylglycerol matrix of the oil body, but the conformation adopted by this sequence is uncertain . We have therefore expressed an oleosin cDNA from sunflower (Helianthus annuus L.) in Escherichia coli as a fusion with maltose-binding protein (MBP) and isolated a peptide corresponding to the hydrophobic domain by sequential digestion with factor Xa (to remove the MBP) followed by trypsin and Staphylococcus V8 protease to remove the N- and C-terminal domains of the oleosin . Circular dichroism spectroscopy of the peptide in two solvent systems chosen to mimic the environment within the oil body (trifluoroethanol and SDS) demonstrated high proportions of alpha-helical structure, with no beta-sheet . A model was therefore developed in which the domain forms an alpha-helical hairpin structure, the two helices being separated by a turn region . We consider that this model is consistent with our current knowledge of oleosin structure and properties.

J Clin Microbiol, 2002 Apr, 40(4), 1333 - 8
rpoB gene sequence-based identification of Staphylococcus species; Drancourt M et al.; The complete sequence of rpoB, the gene encoding the beta subunit of RNA polymerase was determined for Staphylococcus saccharolyticus, Staphylococcus lugdunensis, S taphylococcus caprae, and Staphylococcus intermedius and partial sequences were obtained for an additional 27 Staphylococcus species . The complete rpoB sequences varied in length from 3,452 to 3,845 bp and had a 36.8 to 39.2% GC content . The partial sequences had 71.6 to 93.6% interspecies homology and exhibited a 0.08 to 0.8% intraspecific divergence . With a few exceptions, the phylogenetic relationships inferred from the partial rpoB sequences were in agreement with those previously derived from DNA-DNA hybridization studies and analyses of 16S ribosomal DNA gene sequences and partial HSP60 gene sequences . The staphylococcal rpoB sequence database we established enabled us to develop a molecular method for identifying Staphylococcus isolates by PCR followed by direct sequencing of the 751-bp amplicon . In blind tests, this method correctly identified 10 Staphylococcus isolates, and no positive results were obtained with 10 non-Staphylococcus gram-positive and gram-negative bacterial isolates . We propose partial sequencing of the rpoB gene as a new tool for the accurate identification of Staphylococcus isolates.

Clin Exp Metastasis, 2002, 19(1), 43 - 53
Synergistic effect of interleukin-2 and a vaccine of irradiated melanoma cells transfected to secrete staphylococcal enterotoxin A; Schrayer DP et al.; We have previously reported that immunization of mice with melanoma cells transfected to secrete the superantigen, Staphylococcal enterotoxin A (SEA), increased the production of antibodies to the B700 melanoma antigen, stimulated the production of endogenous interleukin 2 (IL-2), activated the expression of CD4, CD8 and CD25 T cell markers and enhanced NK cell activity . Now we show that immunization of mice with a vaccine of irradiated sea-transfected melanoma cells coupled with IL-2 therapy was even more effective in inhibiting the growth of primary melanoma tumors and the development of lung metastases than was the irradiated melanoma cell vaccine alone or IL-2 alone . The morphological and immunological effectiveness of the therapy was dose-dependent on IL-2.

Ann Fr Anesth Reanim, 2002 Feb, 21(2), 78 - 83
{Cerebrospinal fluid shunting: anesthetic particularities}; Sesay M et al.; The symptomatic treatment of hydrocephalus remains cerebrospinal fluid (CSF) drainage to an external reservoir (external CSF drainage) or to an internal cavity mainly the peritoneum or the right atrium via a unidirectional valve (internal CSF drainage) and finally by endoscopic ventriculocisternostomy . Local anaesthesia is adequate for external CSF drainage in adults and children above 10 years while general anaesthesia is required in all other cases . The main problems encountered in these patients are difficult intubation and full stomach associated with increased intracranial pressure . The anaesthetic approach should favour homeostasis . With the exception of ketamine and enflurane, the majority of anaesthetic drugs can be used . Anti-epileptic drug are mandatory . Antibioprophylaxis mainly against staphylococcus is systematic in internal CSF drainage . Rapid emergence from anaesthesia and extubation should be encouraged . Complications (infectious, mechanical and bleeding kinds) are frequent and are often the cause of reinterventions or revisions of the device, exposing the patients to iterative anaesthesia . Furthermore, patients with shunts are at risk of malfunction of the device when exposed to situations like pregnancy, magnetic resonance imaging, or laparoscopy . Under these circumstances, it is recommended to associate the neurosurgical team in the management of these patients and to verify that the shunt is working well before and after the procedure or event.

Blood Rev, 2002 Mar, 16(1), 73 - 6
HIV-related thrombocytopenia; Scaradavou A; Chronic thrombocytopenia is a common hematologic disorder in patients infected with the human immunodeficiency virus (HIV) . Although often asymptomatic, the thrombocytopenia may be associated with a variety of bleeding abnormalities . The underlying pathophysiology includes accelerated peripheral platelet destruction and decreased ('ineffective') production of platelets from the infected megakaryocytes . HIV-related thrombocytopenia (HIV-TP) responds to antiretroviral therapy . Most studies have evaluated the use of AZT (zidovudine) and have shown increased platelet production . Combination therapy (HAART) also resulted in sustained platelet increases . When antiretroviral agents fail to improve the platelet count or cannot be used, other therapies, similar to those used in 'classic' immune thrombocytopenia (ITP), can be employed, including steroids, intravenous immunoglobulin (i.v.intravenous anti-D or splenectomy . Anti-D treatment offers advantages for HIV-TP because the duration of effect appears to be significantly longer than the response duration after i.v . therapy (initial results of our open-arm study were confirmed by our randomized trial) . Of note, follow-up of heavily treated patients showed no acceleration of CD4 decline and no change in plasma viral load measurements . Splenectomy has been used to treat HIV-positive patients with refractory thrombocytopenia . Although it is effective therapy, there are concerns about infections and selection of appropriate candidates . Other treatment modalities, such as interferon, vincristine, danazol, low-dose splenic irradiation and staphylococcal protein A immunoadsorption have shown limited success in HIV-TP . Alternatively, thrombocytopenia in HIV-infected patients may be treated with pharmacological hyperstimulation of megakaryocytopoiesis (administration of PEG-rHuMGDF or TPO) . Latest evidence indicates that the chemokine receptor CXCR4 (coreceptor for the cellular entry of lymphotropic HIV strains) is expressed on megakaryocytes; as a result, the development of chemokine receptor antagonists may modify the course of the disease .

J Pediatr Surg, 2002 Apr, 37(4), 673 - 4
Meconium thorax: A case of Bochdalek hernia and cecal perforation in a neonate with Job's syndrome; Butterworth SA et al.; Meconium peritonitis most often is associated with congenital intestinal obstruction and meconium ileus . Uncommonly, other etiologies are identified . Hyperimmunoglobulin E syndrome (Job's syndrome) is a rare genetic disorder that is characterized by recurrent staphylococcal respiratory and skin infections in addition to elevated serum IgE levels . There have been 2 previously reported cases of intestinal complications associated with Job's syndrome . The current case is the third such case and is the first report of meconium peritonitis in a patient with hyperimmunoglobulin E syndrome . The patient presented with a meconium thorax as a result of a concurrent congenital diaphragmatic hernia .

Protein Sci, 2002 Apr, 11(4), 894 - 902
Subunit composition of a bicomponent toxin: staphylococcal leukocidin forms an octameric transmembrane pore; Miles G et al.; Staphylococcal leukocidin pores are formed by the obligatory interaction of two distinct polypeptides, one of class F and one of class S, making them unique in the family of beta-barrel pore-forming toxins (beta-PFTs) . By contrast, other beta-PFTs form homo-oligomeric pores; for example, the staphylococcal alpha-hemolysin (alpha HL) pore is a homoheptamer . Here, we deduce the subunit composition of a leukocidin pore by two independent methods: gel shift electrophoresis and site-specific chemical modification during single-channel recording . Four LukF and four LukS subunits coassemble to form an octamer . This result in part explains properties of the leukocidin pore, such as its high conductance compared to the alpha HL pore . It is also pertinent to the mechanism of assembly of beta-PFT pores and suggests new possibilities for engineering these proteins.

J Antimicrob Chemother, 2002 Apr, 49(4), 693 - 6
An in vitro evaluation of the antibiotic/heparin lock to sterilize central venous haemodialysis catheters; Vercaigne LM et al.; This in vitro study investigated the ability of antibiotic/heparin locks to sterilize central venous haemodialysis catheters (CVCs) inoculated with methicillin-resistant Staphylococcus epidermidis (MRSE) . Isolates of MRSE were incubated in broth inside CVCs . The catheters were then drained and filled with either vancomycin/gentamicin/heparin (VGH), cefazolin/gentamicin/ heparin (CGH) or control locks for 48 h . The catheters were drained, filled with fresh broth and again incubated . The final catheter solutions were sampled and the remaining volumes filtered . The samples, filters and catheter segments were examined for growth . For two isolates, both the VGH and CGH locks sterilized the catheters . Bacterial counts of the remaining two isolates were significantly reduced by >99%, but the catheters were not sterilized after the instillation of a single-antibiotic/heparin lock.

Nature, 2002 Mar 21, 416(6878), 291 - 7
Killing activity of neutrophils is mediated through activation of proteases by K+ flux; Reeves EP et al.; According to the hitherto accepted view, neutrophils kill ingested microorganisms by subjecting them to high concentrations of highly toxic reactive oxygen species (ROS) and bringing about myeloperoxidase-catalysed halogenation . We show here that this simple scheme, which for many years has served as a satisfactory working hypothesis, is inadequate . We find that mice made deficient in neutrophil-granule proteases but normal in respect of superoxide production and iodinating capacity, are unable to resist staphylococcal and candidal infections . We also show that activation provokes the influx of an enormous concentration of ROS into the endocytic vacuole . The resulting accumulation of anionic charge is compensated for by a surge of K+ ions that cross the membrane in a pH-dependent manner . The consequent rise in ionic strength engenders the release of cationic granule proteins, including elastase and cathepsin G, from the anionic sulphated proteoglycan matrix . We show that it is the proteases, thus activated, that are primarily responsible for the destruction of the bacteria.

Biochem J, 2002 Apr 1, 363(Pt 1), 175 - 82
Opposing roles of serine/threonine kinases MEKK1 and LOK in regulating the CD28 responsive element in T-cells; Tao L et al.; T-cell activation requires signals from both the T-cell receptor (TcR) and other co-stimulatory molecules such as CD28 . TcR- and CD28-mediated signals are integrated during T-cell activation resulting in the expression of cytokine genes such as interleukin-2 (IL-2) . An enhancer element (CD28RE) of the IL-2 gene specifically responsive to CD28 signals has been previously identified and characterized . This response element and an adjacent Activated Protein-1 (nuclear factor-interleukin-2B) site together (RE/AP1) were shown to complex with c-rel, AP-1 and other factors . However, details of the signal transduction pathways leading from CD28 to the composite response element remain poorly understood . We present data showing that overexpression of the serine threonine kinase, mitogen-activated protein kinase/extracellular-signal-regulated kinase kinase kinase-1 (MEKK1), but not nuclear factor-kappa B inducing kinase, or MAP kinase/ERK kinase-1 (MEK1), can significantly increase the level of CD28RE/AP1-driven luciferase (Luc) reporter gene expression in Jurkat E6-1 cells . A MEKK1 dominant negative mutant blocked such activation induced by stimulation with Raji B cells and the superantigen staphylococcus enterotoxin E (SEE), as well as via CD3/CD28 . Mutations in either site of the RE/AP1 element abolished MEKK1-induced Luc expression . Calcineurin inhibitors, CsA and FK520, or inhibitors of p38 kinase (SB 203580), or MEK1 (PD 098059), did not affect MEKK1-induced reporter activation . These results directly implicate MEKK1 in the CD28 signalling pathway that activates the CD28 response element . Co-expression of the lymphocyte-oriented kinase (LOK) kinase attenuated Raji/SEE-induced IL-2 production in Jurkat cells, as well as MEKK1 and Raji/SEE-induced reporter gene activation . These data suggest that MEKK1 and LOK may have opposing roles in regulating the CD28RE/AP1 element.

Cell Biochem Biophys, 2001, 34(3), 283 - 304
Determining the three-dimensional fold of a protein from approximate constraints: a simulation study; Soman KE et al.; We propose a new approach for calculating the three-dimensional (3D) structure of a protein from distance and dihedral angle constraints derived from experimental data . We suggest that such constraints can be obtained from experiments such as tritium planigraphy, chemical or enzymatic cleavage of the polypeptide chain, paramagnetic perturbation of nuclear magnetic resonance (NMR) spectra, measurement of hydrogen-exchange rates, mutational studies, mass spectrometry, and electron paramagnetic resonance . These can be supplemented with constraints from theoretical prediction of secondary structures and of buried/exposed residues . We report here distance geometry calculations to generate the structures of a test protein Staphylococcal nuclease (STN), and the HIV-1 rev protein (REV) of unknown structure . From the available 3D atomic coordinates of STN, we set up simulated data sets consisting of varying number and quality of constraints, and used our group's Self Correcting Distance Geometry (SECODG) program DIAMOD to generate structures . We could generate the correct tertiary fold from qualitative (approximate) as well as precise distance constraints . The root mean square deviations of backbone atoms from the native structure were in the range of 2.0 A to 8.3 A, depending on the number of constraints used . We could also generate the correct fold starting from a subset of atoms that are on the surface and those that are buried . When we used data sets containing a small fraction of incorrect distance constraints, the SECODG technique was able to detect and correct them . In the case of REV, we used a combination of constraints obtained from mutagenic data and structure predictions . DIAMOD generated helix-loop-helix models, which, after four self-correcting cycles, populated one family exclusively . The features of the energy-minimized model are consistent with the available data on REV-RNA interaction . Our method could thus be an attractive alternative for calculating protein 3D structures, especially in cases where the traditional methods of X-ray crystallography and multidimensional NMR spectroscopy have been unsuccessful.

Rev Esp Cardiol, 2002 Mar, 55(3), 304 - 7
{Risk factors associated with endocarditis without underlying heart disease}; Castillo JC et al.; Infective endocarditis (IE) pathogenesis has changed in the last decades and there is an increasing number of patients without predisposing heart condition . The aim of this study is to asses the clinical features of these non-drug addict patients affected with IE without underlying heart disease and to identify the potential risk factors . From 196 cases of IE, 49 (25% of the series) occurred in patients without underlying heart disease . A presumed portal of entry was identified in the majority (26 cases) . The most frequent were digestive (6 cases), haemodialysis (6 cases) and central venous catheters (4 cases) . Right heart valves were more often affected (29 vs 6%; p < 0.01) . The distribution of the causative microorganism showed a higher proportion of Staphylococcus (57 vs 30%) . Despite a similar in-hospital complication rate and a similar need of surgery during the active phase, their prognosis is better than in those with underlying heart disease.

Enferm Infecc Microbiol Clin, 2002 Feb, 20(2), 74 - 7
{Infections in joint prostheses: epidemiology and clinical presentation . A prospective study 1992-1999}; Gomez J et al.; BACKGROUND: The prosthetic infection is a serious complication due to diagnostic problems . AIM: To determinate epidemiological characteristics, and clinical patterns of infections associated to prosthetic materials to improve the diagnosis and management . PATIENTS AND METHODS: From december 1992 to december 1999, 110 patients with prosthetic infections were prospectively evaluated . Diagnosis was made according to standard microbiological, clinical and radiological criteria . RESULTS: The incidence was 5.1% (110/1,400) prosthetic materials insert in the period of study . The average age was 59.6 years (range 18-79), and the majority of patients 63 (57.2%) were female . Forty-two (38%) suffered a total knee replacement, 29 (26%) a total hip replacement, 1 (1%) shoulder replacement and 38 (34%) autogenous bone gratting . In 29 patients (26.3%) a previous chronic disease had been diagnosed (diabetes, neoplasis, rheumatoid arthritis) . Previous use of antibiotics was detected in 58 patients (51%), being ciprofloxacin the most frequently used . An etiological diagnosis was reached in 66 patients (60%), isolated grampositive in 58.2%, gramnegative in 32.8%, fundamentally by Staphylococcus sp . and P . aeruginosa respectively . In 9% anaerobe were isolated . There were early infections in 67 cases, delayed in 25, and late in 18 . All the patients had local pain and flogotiv signs as initial findings, whereas 46 (41.8%) developed osteocutaneous fistula and only 5 (4.5%) presented temperature . CONCLUSIONS: Prosthetic infection is a frequent complication after articular replacement, and grampositive cocci predominate as ethilogical agents . Sistemic clinical manifestations are uncommon.

Clin Experiment Ophthalmol, 2002 Apr, 30(2), 144 - 6
Septic superior ophthalmic vein thrombosis; Walker JC et al.; An illustrative case of septic superior ophthalmic vein thrombosis secondary to a staphylococcus orbital cellulitis is presented and correlated with autopsy findings . A literature review and discussion of the risks and benefits of anti-coagulation in this setting is outlined.

J Hosp Infect, 2002 Mar, 50(3), 183 - 7
The effect of 10% povidone--iodine solution on contaminated bone allografts; Soyer J et al.; The aim of this study was to determine the efficacy of 10% povidone--iodine solution for the decontamination of bone allografts . Bone samples were prepared and tested for sterility using a femoral head removed at the time of primary hip replacement . They were contaminated by a suspension of Staphylococcus epidermidis and ground to measure the quantity of micro-organism attached to the bone . Two levels of contamination were used (1 x 10(3) vs . 1 x 10(4)CFU/mL) to check the efficiency of our method of measurement . Samples of the two groups were decontaminated with 10% povidone--iodine solution using different exposure times . Before decontamination, the count of bacteria attached to the bone was proportional to the bacterial concentration of the contaminating solution . The microbiocidal activity of 10% povidone--iodine solution was the same in both groups . The decontamination time was proportional to the bacterial concentration of the contaminating solution . The results of this preliminary study suggest that a 10% povidone--iodine solution can decontaminate inoculated bone grafts, but a sufficient time of exposure according to the level of contamination must be allowed .

Haemophilia, 2002 Jan, 8 Suppl 1, 20 - 3; discussion 28-32
Use of Protein-A column and porcine factor VIII; Rivard GE; Extracorporeal immunoadsorption of factor VIII (FVIII) antibodies using Sepharose matrix columns coupled with staphylococcal Protein-A was reported two decades ago . The efficiency of this technique for removing FVIII antibodies of the IgG subtypes was clearly demonstrated . The recent widespread use of a variety of apheresis techniques for the management of a multitude of haematological and oncological conditions has made this technology more accessible and affordable . For the treatment of patients with FVIII inhibitors, the use of porcine FVIII makes it possible to control haemostasis with a therapeutic product for which in vitro testing can help predict the in vivo efficacy . By lowering the level of FVIII inhibitors, immunoadsorption can make the use of pFVIII concentrate possible in situations otherwise untreatable with FVIII preparations . Moreover, lowering the level of FVIII inhibitors by immunoadsorption allows adequate haemostasis to be achieved with much lower doses of FVIII leading to significant saving . Our preliminary data suggest that immunoadsorption combined with the use of pFVIII should be considered early in the treatment plan for controlling haemostasis in patients with FVIII inhibitors.

J Clin Microbiol, 2002 Mar, 40(3), 1109 - 12
Staphylococcus heterogeneously resistant to vancomycin in China and antimicrobial activities of imipenem and vancomycin in combination against it; Benquan W et al.; Of 115 methicillin-resistant Staphylococcus strains collected from sputum specimens, 34 strains reduced susceptibility to vancomycin, 9 of which emerged as heterogeneous vancomycin-resistant strains (hetero-VRS), with various degrees of vancomycin resistance at a frequency of 10(-6) or higher . Seventy-six percent (19 of 25) of non-hetero-VRS and 100% (9 of 9) of hetero-VRS were susceptible to synergistic treatment with vancomycin and imipenem . Clinical clearance between 9 hetero-VRS and 25 non-hetero-VRS had an obvious statistical significance (P = 0.001) . The hetero-VRS may play an important role in vancomycin therapy failure.

J Clin Microbiol, 2002 Mar, 40(3), 898 - 901
Initial concentration of Staphylococcus epidermidis in simulated pediatric blood cultures correlates with time to positive results with the automated, continuously monitored BACTEC blood culture system; Haimi-Cohen Y et al.; The relationship of initial concentration of Staphylococcus epidermidis in blood cultures and time to positivity (TTP) in an automated, continuously monitored blood culture system was assessed . Blood and 1 to 1,000 CFU of S . epidermidis per ml in stationary or exponential phase were inoculated in BACTEC Pediatric Plus F bottles and incubated . The TTP was inversely proportional to the initial inoculated concentration . Blood culture bottles with initial bacterial densities of <10 CFU/ml had a TTP of >20 h (upper limit of 95% prediction interval, 20.7 h) and bottles with initial bacterial densities of > or =50 CFU/ml had a TTP of < or =15 h (lower limit of 95% prediction interval, 13.4 h).

J Neuroimmunol, 2002 Feb, 123(1-2), 41 - 9
Effects of staphylococcal enterotoxin A on pituitary-adrenal activation and neophobic behavior in the C57BL/6 mouse; Kawashima N et al.; Bacterial superantigens, such as the staphylococcal enterotoxins (SE), exert a strong capacity for in vivo stimulation of T cell proliferation and cytokine production . Among these superantigens, staphylococcal enterotoxin B (SEB) has been shown to promote anxiety-like properties, possibly mediated by activation of central corticotropin-releasing hormone . In the present study, using male C57BL/6J mice, it was shown that challenge with another prominent superantigen, staphylococcal enterotoxin A (SEA), produced a dose-dependent (0.2-50 microg/mouse) increase in corticosterone and ACTH levels . Interestingly, while the adrenocorticoid response to SEA persisted in cyclosporine A-pretreated mice, it was completely abolished in RAG-1 deficient mice that lack functional T and B lymphocytes . The latter is consistent with the need for cellular interactions involving T cells and B cells (probably in an antigen-presenting capacity) that will initiate events leading to pituitary-adrenal activation by SEA . Since pituitary-adrenal activation typically alters "emotional" reactivity in animals, a final set of experiments assessed behavioral responses to an open field, exposure to a novel object, and a novel appetitive stimulus . These tests revealed a significant augmentation of reactivity to the novel object in SEA-challenged mice, although activity in the open field was not affected . Furthermore, consumption of a novel solution was reduced only if testing involved unfamiliar contextual circumstances . This suggested that anorexic effects per se were not induced by SEA at the dose used, but that attentional mechanisms focused on novelty were enhanced.

J Neuroimmunol, 2002 Feb, 123(1-2), 30 - 4
Intramolecular epitope spreading induced by staphylococcal enterotoxin superantigen reactivation of experimental allergic encephalomyelitis; Soos JM et al.; The staphylococcal enterotoxin superantigens are among the most potent T cell stimulators known . They have been shown to alter the course of disease in experimental allergic encephalomyelitis, an animal model for multiple sclerosis (MS) . We have previously demonstrated that two of the staphylococcal enterotoxins, SEA and SEB, are able to reactivate paralysis in PL/J mice which had been immunized with myelin basic protein (MBP) and resolved an initial episode of paralysis . In PL/J mice, Ac1-11 is the dominant encephalitogenic determinant of MBP . We hypothesized that superantigen reactivation of experimental allergic encephalomyelitis (EAE) may result in the spreading of T cell specificities for other epitopes of MBP . PL/J mice which had resolved an initial episode of EAE were treated with SEA and developed a second episode of paralysis . At the onset of symptoms, mice were sacrificed and splenocytes were stimulated in vitro with a panel of MBP peptides . EAE reactivation by SEA resulted in the spreading of T cell specificites to residues 100 to 120 of MBP . While intramolecular spreading did occur, spreading to other antigens did not, as evidenced by the lack of response to a proteolipid protein (PLP) peptide and heat shock protein 60 (hsp 60) . To further characterize the epitope MBP 100-120, PL/J mice were immunized with MBP 100-120 . No initial development of disease was observed . However, administration of SEA 2 weeks after MBP 100-120 immunization resulted in the onset of paralysis . In addition to a proliferative response to MBP 100-120, these mice also exhibited a proliferative response to the flanking MBP peptides 81-100 and 120-140 . Thus, SEA is able to induce intramolecular epitope spreading in PL/J mice after reactivation of EAE.

Mil Med, 2002 Feb, 167(2), 157 - 60
Magnetic resonance imaging findings in a case of cytarabine-induced myelopathy; Sherman PM et al.; A 44-year-old white male with an isolated central nervous system relapse of acute lymphoblastic leukemia was treated with intrathecal cytarabine . He developed Staphylococcus epidermidis meningitis, which was treated successfully with intrathecal vancomycin . Four weeks after the initiation of intrathecal cytarabine, the patient developed progressive ascending paralysis to the upper cervical level . Initial magnetic resonance imaging of the brain and spine were normal, and cerebrospinal fluid evaluation showed no evidence of ongoing infection and clearance of lymphoblasts . Three weeks later, magnetic resonance imaging demonstrated marked edema and peripheral enhancement of the spinal cord, consistent with cytarabine toxicity.

J Appl Microbiol, 2002, 92(3), 482 - 90
Proteolytic activity of Staphylococcus xylosus strains on pork myofibrillar and sarcoplasmic proteins and use of selected strains in the production of "Naples type" salami; Mauriello G et al.; AIMS: The aim of this study was to determine the proteolytic activities of Staphylococcus xylosus strains on sarcoplasmic and myofibrillar proteins in order to evaluate the suitability of selected strains as starter cultures in the processing of a dry fermented pork sausage . METHODS AND RESULTS: The proteolytic activity of 27 strains of Staphylococcus xylosus on sarcoplasmic and myofibrillar proteins was determined by agar plate method, o-phtaldialdehyde (OPA) spectrophotometric assay and sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) . Four strains were selected for the formulation of six starter cultures to use in the production of "Naples type" salami . The proteolytic contribution of starters was determined by SDS-PAGE, comparing the protein profile of inoculated sausages with that of uninoculated sausages after 0, 15 and 33 days of ripening . The results showed that the proteolytic activity of some strains, determined by the agar plate method, were not confirmed by electrophoretic and spectrophotometric assays . In fact, of 24 strains of Staphylococcus xylosus able to hydrolyse muscle protein extracts on agar plate, only 12 strains were shown to change SDS-PAGE profile of pork proteins . The SDS-PAGE profile of sarcoplasmic proteins extracted from all sausages showed that the major changes were produced with starters S3, S4 and S5 after 15 days of ripening . Also myofibrillar proteins undergo major changes after 15 days of ripening and the protein profiles showed the same pattern in all samples, except for the sausages produced with starter S4 . CONCLUSIONS: The results of this work showed that the muscle protein extracts hydrolysis test is suitable for preliminary screening of Staphylococcus xylosus strains on the basis of their proteolytic activity . However, evaluation of muscle protein hydrolysis in a food model system could then be more appropriate for selecting micro-organisms for use as starter cultures for fermented sausages . SIGNIFICANCE AND IMPACT OF THE STUDY: The potential of the findings is discussed with reference to the formulation of starter cultures for the dry fermented sausages production.

Immunology, 2002 Feb, 105(2), 163 - 70
Induction of thymocyte positive selection does not convey immediate resistance to negative selection; Hare KJ et al.; The acquisition of functional competence represents a critical phase during intrathymic development of T cells . Thymocytes reaching this stage represent cells which have been positively selected on the basis of major histocompatibility complex reactivity, but which have also been purged of potentially autoreactive T-cell receptor specificities by negative selection . While the developmental window in which thymocytes are subjected to positive selection is now well defined, the precise developmental timing of negative selection, in relation to positive selection events, is less clear . Moreover, the underlying mechanism allowing single-positive thymocytes to respond to T-cell receptor ligation by activation rather than death, remains controversial . Here we have analysed the developmental timing of negative selection in relation to positive selection, using measurement of thymocyte susceptibility to dendritic cell presentation of the superantigen staphylococcal enterotoxin B (SEB) . We show that thymocytes which have received initial positive selection signals, namely CD4+ CD8+ CD69+ thymocytes, like their CD4+ CD8+ CD69minus sign precursors, are susceptible to negative selection, indicating that induction of positive selection does not convey immediate resistance to negative selection . In contrast, newly generated CD4+ CD8minus sign CD69+ cells are not only resistant to deletion by SEB, but respond to SEB-mediated T-cell receptor-ligation by activation, indicating that the acquisition of functional competence occurs at the newly generated CD4+ CD8minus sign CD69+ stage . Finally, by using direct retroviral infection of primary CD4+ CD8+ thymocytes, we also show that Notch-1 activation in CD4+ CD8+ thymocytes does not correlate with, nor convey resistance to superantigen-mediated negative selection . Thus, our data suggest that although Notch-1 has been implicated in resistance to thymocyte apoptosis, the acquisition of resistance to negative selection occurs independently of Notch-1 signalling.

JPEN J Parenter Enteral Nutr, 2002 Mar-Apr, 26(2), 109 - 13
Efficacy of antibiotic-lock technique with teicoplanin in staphylococcus epidermidis catheter-related sepsis during long-term parenteral nutrition; Guedon C et al.; BACKGROUND: The antibiotic-lock technique has been suggested to treat catheter-related sepsis in parenteral nutrition and to avoid catheter removal . METHODS: To determine the incidence of catheter-related sepsis, the bacteria involved, and the efficacy of the antibiotic-lock technique with teicoplanin, all patients (n = 263) undergoing parenteral nutrition from January 1997 to December 1999 in one center, with patients at the hospital (n = 209) and at home (54) were retrospectively studied . The antibiotic-lock technique with teicoplanin was systematically used in all suspected infections and maintained in staphylococcus epidermidis (SE) infections . RESULTS: A total of 21 of 263 patients had 34 infections (0.11/patient per year): 12 of 209 hospitalized and 9 of 54 home patients . A total of 10 of 34 infections were due to non-SE, and the catheter was immediately removed . The other 24 of 34 infections were due to SE; in 5 of 24, the catheter was removed after 48 hours of the antibiotic-lock technique because of persistent fever or thrombosis . A total of 5 of 12 patients had 2 or more infections on the same catheter . The antibiotic-lock technique prevented short-term catheter removal in these cases, but a second infection occurred within a median of 50 days . In 4 of 5 cases, a third infection occurred in a mean delay of 90 days so that the catheter was removed . In 3 of 5 patients, bacteria was analyzed with pulsed field gel electrophoresis, which showed that recurrent infections were due to the same strain in all cases . CONCLUSIONS: In this study, the incidence of catheter-related sepsis was low and mostly related to SE . Our results do not support the use of the teicoplanin antibiotic-lock technique in SE infections.

Am Fam Physician, 2002 Feb 15, 65(4), 663 - 70
Linezolid: its role in the treatment of gram-positive, drug-resistant bacterial infections; Ament PW et al.; While the choices available for the management of gram-positive, drug-resistant bacterial infections are becoming limited, antimicrobial resistance is becoming increasingly problematic because of the widespread overuse of antibiotics . Linezolid is a synthetic antibiotic belonging to a new class of antimicrobials called the oxazolidinones . Linezolid disrupts bacterial growth by inhibiting the initiation process of protein synthesis--a mechanism of action that is unique to this class of drugs . It is well absorbed with high bioavailability that allows conversion to oral therapy as soon as the patient is clinically stable . It has been approved for certain gram-positive infections including certain drug-resistant enterococcus, staphylococcus, and pneumococcus strains . It is generally well tolerated, with myelosuppression being the most serious adverse effect . As a nonselective inhibitor of monoamine oxidase, caution is recommended when used with adrenergic or serotonergic agents (e.g., tyramine, dopamine, pseudoephedrine, and selective serotonin reuptake inhibitors) . Judicious use of this medication should help physicians treat patients with multidrug-resistant infections.

Lancet Infect Dis, 2001 Oct, 1(3), 175 - 88
Systematic review and meta-analysis of antibiotic therapy for bone and joint infections; Stengel D et al.; We set out to evaluate the clinical efficacy of individual antibiotic agents for bone and joint infections in adults . Published and unpublished controlled trials reported between 1966 and 2000 were reviewed to determine if they involved random or quasi-random allocation to systemically administered antimicrobials or local antibiotic therapy for osteomyelitis and septic arthritis . Quiescence of infection after 1 year of follow-up was defined as the primary outcome measure . 22 trials containing 927 patients were eligible for final analysis . Varying proportions of the entire study population could be evaluated with respect to primary and secondary endpoints . Methodological quality was poor among most studies, and interpretability of results was further limited by small sample sizes, missing descriptions of patient populations and disease characteristics, and the frequent application of concomitant antibiotics . A trend towards improved, long-lasting infection control was observed in favour of a rifampicin-ciprofloxacin combination versus ciprofloxacin monotherapy for the treatment of staphylococcal infections related to orthopaedic devices (absolute risk difference {ARD} 28-9%; 95% CI -0.7 to 54.4%) . Obviously unbalanced comparative studies showed some benefit of ticarcillin for bone infections caused by Pseudomonas species . No significant differences in therapeutic efficacy were found among trials comparing oral fluoroquinolones with intravenous beta-lactam drugs for both end-of-treatment (OR 0.8; 0.5 to 1.4) and long-term results (OR 1.3; 0.8 to 2.1) . A variety of drugs was used as controls, thereby leading to inconsistent findings of drug-related side effects . Only one randomised trial was suitable to investigate the impact of polymethylmethacrylate gentamicin bead chains compared with parenteral antibiotics for skeletal infections, although this study was biased by patients receiving both combined local and systemic antibiotic therapy . Whereas intention-to-treat evaluation suggested a therapeutic advantage of systemic over local therapy, this trend diminished in the per-protocol analysis (1-year follow-up ARD -2.3;-17.5 to 10.8%) . There exists little high-quality evidence on antibiotic therapy for osteomyelitis and septic arthritis . The observed heterogeneity among patient populations and medical and surgical treatment concepts preclude reliable inferences from the available data.

Zh Mikrobiol Epidemiol Immunobiol, 2001 Sep-Oct, (5), 42 - 5
{Correction of immune response using purified staphylococcal toxoid and likopid in the secondary immunodeficiency induced by Coxsackie virus B3}; Semenova IB et al.; The action of immunomodulators, purified staphylococcal toxoid (PST) and lycopid, on secondary immunodeficiency state developing during infection caused by Coxsackie virus B3 was studied . This defect was manifested by delayed hypersensitivity to sheep red blood cells (SRBC) and the suppression of neutralizing antibodies to poliomyelitis virus . Depending on the scheme of the experiment, PST normalized the defects of immune response to SRBC or poliovirus, increased suppression or showed no activity . Lycopid corrected the defects of humoral response to SRBC . The combination of PST and lycopid was found to produce no increase of suppression . The suggestion was made on the expediency of combination of two (and probably more) immunomodulators for increasing the efficiency of correction of secondary immunodeficiency.

Scand J Infect Dis, 2001, 33(12), 930 - 2
Intravenous catheter-related septic shock caused by Staphylococcus sciuri and Escherichia vulneris; Horii T et al.; Staphylococcus sciuri and Escherichia vulneris were isolated concurrently in a blood sample from a patient with septic shock, which was probably associated with an indwelling catheter . Our results also showed that S . sciuri is an important reservoir of genetic determinants of beta-lactam resistance as a human pathogen carrying the mecA and beta-lactamase genes.

Clin Microbiol Infect, 1996 Aug, 2(1), 44 - 49
Multiresistant Staphylococcus epidermidis in a neonatal care unit; Sloos JH et al.; OBJECTIVE: To study the presence and diversity of types of Staphylococcus epidermidis in the neonatal intensive care unit of a university hospital . METHODS: During a period of 6 weeks, samples were taken from nose, external auditory canal, axilla, groin and umbilicus from consecutively admitted patients . Patients were sampled two times a week for up to 2 weeks . Isolates of S . epidermidis were characterized by antibiogram, plasmid pattern and biotype . RESULTS: Fifteen patients were included . Each patient was sampled in one to four successive surveys, depending on the admission period . A total of 128 isolates of S . epidermidis were obtained and allocated to seven antibiogram types, 36 plasmid types and 14 biotypes . One plasmid type found in 58 isolates (six patients) corresponded with one multiresistant antibiogram type . The number of isolates with these characteristics increased per neonate from the first survey to the fourth . Nineteen isolates from four patients were allocated to a second plasmid type and were of a common antibiogram type . The remaining 34 plasmid types were sporadic . No clear correspondence of biotypes with antibiogram or plasmid types was found . CONCLUSIONS: The present study revealed the increase in colonization of a multiresistant type of S . epidermidis in the compromised patients during admission to the ward . Further studies have to assess whether this type remains persistent in the ward.

Anal Chem, 2002 Feb 15, 74(4), 841 - 7
Conjugation of luminescent quantum dots with antibodies using an engineered adaptor protein to provide new reagents for fluoroimmunoassays; Goldman ER et al.; We describe the preparation and characterization of bioinorganic conjugates made with highly luminescent semiconductor CdSe-ZnS core-shell quantum dots (QDs) and antibodies for use in fluoroimmunoassays . The conjugation strategy employs an engineered molecular adaptor protein, attached to the QDs via electrostatic/hydrophobic self-assembly, to link the inorganic fluorophore with antibodies . In this method, the number of antibodies conjugated to a single QD can be varied . In addition, we have developed a simple purification strategy based on mixed-composition conjugates of the molecular adaptor and a second two-domain protein that allows the use of affinity chromatography . QD-antibody conjugates were successfully used in fluoroimmunoassays for detection of both a protein toxin (staphylococcal enterotoxin B) and a small molecule (2,4,6-trinitrotoluene).

J Infect Dis, 2002 Feb 15, 185(4), 555 - 60 Epub 2002 Jan 22.
Temporal sequence and functional implications of V beta-specific T cell receptor down-regulation and costimulatory molecule expression following in vitro stimulation with the staphylococcal superantigen Toxic shock syndrome toxin-1; Kum WW et al.; The superantigen toxic shock syndrome toxin-1 (TSST-1) is implicated as the major cause of staphylococcal toxic shock syndrome . The temporal sequence of early signaling events in human peripheral blood mononuclear cells following TSST-1 stimulation was examined . TSST-1 induced rapid and complete down-regulation of V beta 2-specific T cell receptor (TCR), followed by transient CD154 expression on CD4(+) lymphocytes . This was sequentially followed by the up-regulation of CD86, CD80, CD40, and human leukocyte antigen-DR expression on CD14(+) monocytes . In contrast, S14N, a TSST-1 mutant toxin with a single amino acid substitution that is known to be impaired in interleukin (IL)--2, interferon (IFN)-gamma, and tumor necrosis factor (TNF)-alpha secretion, was deficient in both V beta 2-TCR down-regulation and CD154 and CD80/CD86 expression . Furthermore, pretreatment with monoclonal antibodies against V beta 2-TCR, CD80/CD86, and CD154 significantly inhibited TSST-1-induced IL-2, IFN-gamma, and TNF-alpha secretion . Taken together, these results indicate that early V beta-specific TCR activation, along with CD80/CD86 and CD154 costimulation, are key determinants of the TSST-1-induced proinflammatory cytokine response.

Br J Ophthalmol, 2002 Mar, 86(3), 261 - 5
Risk factors for graft infection in India: a case-control study; Vajpayee RB et al.; AIM: To study the demographic, clinical, and microbiological profile and the risk factors for graft infection following penetrating keratoplasty . METHODS: 50 eyes of 50 consecutive patients with graft infection after an optical penetrating keratoplasty were included as cases; 50 eyes of 50 patients with no graft infection were included as controls . The main variables evaluated in this study included the clinical and microbiological profile, sociodemographic status, suture related problems, persistent epithelial defects, and ocular surface disorders . RESULTS: Cultures were positive in 43 (86%) eyes and Staphylococcus epidermidis (67.4%) was the most common organism isolated . Infection could be resolved with treatment in 37 (74%) eyes . In eight (16%) eyes the graft melted and a repeat penetrating keratoplasty had to be performed . Only 6% of the cases could achieve a best corrected visual acuity of 6/18 or better after resolution of the infection . In multivariate logistic regression analysis persistent epithelial defect (OR (95% CI): 3.0 (1.17 to 8.33)), suture related problems (OR (95% CI): 3.6 (1.39 to 9.25)), and ocular surface disorders (OR (95% CI): 2.4 (0.93 to 6.03)) were found to be statistically significant risk factors for graft infection following an optical penetrating keratoplasty . CONCLUSIONS: Staphylococcus epidermidis is the commonest organism responsible for post-keratoplasty microbial keratitis . Persistent epithelial defects, suture related problems, and ocular surface disorders are the major risk factors predisposing to graft infection.

Clin Microbiol Infect, 1997 Jun, 3(3), 356 - 364
PCR-based DNA fingerprinting of Staphylococcus haemolyticus to investigate nosocomial infections; Noordhoek GT et al.; OBJECTIVE: To apply PCR-based DNA fingerprinting in a clinical microbiology laboratory to investigate nosocomial infections with Staphylococcus haemolyticus . METHODS: DNA fingerprints were generated by PCR on 99 S . haemolyticus isolates using different primer combinations based on ERIC, REP or arbitrarily chosen simple repeat sequences . RESULTS: Primer combinations REP1+(GTC)6 and ERIC1+ERIC2 had sufficient discrimatory power and were chosen to analyze the clinical isolates . DNA fingerprint patterns from strains isolated from the patients nursed in the same hospital ward in the period 1991--94 were approximately 90% similar to each other . One staff member, sampled in 1991, carried a strain with a similar fingerprint . CONCLUSIONS: PCR based DNA fingerprinting is a suitable method to perform in a clinical laboratory . An S . haemolyticus strain appeared to be endemic in the hospital ward and had most probably been transmitted from patient to patient . S . haemolyticus may carry glycopeptide resistance and needs attention as a causative agent of nosocomial infections.

Biochemistry, 2002 Mar 5, 41(9), 3089 - 95
Molecular alignment of denatured states of staphylococcal nuclease with strained polyacrylamide gels and surfactant liquid crystalline phases; Ackerman MS et al.; Residual dipolar couplings reflect the orientation of vectors between pairs of magnetic nuclei relative to a unique set of molecular axes . Thus, unlike NOEs and scalar couplings, dipolar couplings provide access to long-range structural information . A prerequisite for measurement of these NMR parameters is imposition of a weak net alignment, most simply by forcing the macromolecules to tumble in an asymmetric environment that restricts some orientations more than others . In this report, several denatured forms of staphylococcal nuclease are aligned by using compressed and stretched polyacrylamide gels, a nonionic type of lipid bilayer disk or bicelle, and a liquid crystalline phase formed by a cationic lipid . All three types of media can be used at high urea concentrations . While polyacrylamide gels and bicelles produce similar alignment tensors through steric interactions, a liquid crystalline phase of cetylpyridinium bromide aligns denatured nuclease along a different set of axes, presumably through electrostatic effects . The analysis of residual dipolar couplings collected with two different alignment tensors may permit the calculation of ensembles of conformations . The dipolar couplings observed for staphylococcal nuclease denatured with urea, by low pH or by deletion of residues from both termini, suggest that all denatured forms share a common "topology", one which has been shown previously to be native-like . Although SDS/nuclease complexes give sharp and disperse (1)H-(15)N correlation spectra, only small couplings are observed in strained polyacrylamide gels.

Drug Dev Ind Pharm, 2002 Jan, 28(1), 95 - 9
In-vitro study of bacterial adherence to different types of intraocular lenses; Burillon C et al.; The aim of this study was to determine the adherence of Staphylococcus epidermidis to intraocular lenses made of five different biomaterials: polymethylmethacrylate (PMMA), heparinized PMMA, silicone, hydrophilic acrylic, and hydrogel . The extent of bacterial binding was measured by counting . The results were compared using a one-factor variance analysis . Adherence was weakest on hydrogel and strongest on the silicone polymer . Bacterial adherence to the implant surface must therefore depend on the hydrophobicity or hydrophilicity of the biomaterial.

J Immunol, 2002 Mar 1, 168(5), 2188 - 99
Administration of an antigen at a high dose generates regulatory CD4+ T cells expressing CD95 ligand and secreting IL-4 in the liver; Watanabe T et al.; Ags administered orally at a high dose are absorbed in immunogenic forms and perfuse the liver, which raises a question regarding the relevance of hepatic lymphocyte activation to the systemic hyporesponsiveness against the ingested Ag . Oral administration of 100 mg of OVA to the mice led to massive cell death of OVA-specific (KJ1-26+)CD4+ T cells by Fas-Fas ligand (FasL)-mediated apoptosis in the liver, which was associated with the emergence of hepatic KJ1-26+CD4+ T cells expressing FasL . Hepatic CD4+ T cells in OVA-fed mice secreted large amounts of IL-4, IL-10, and TGF-beta(1) upon restimulation in vitro and inhibited T cell proliferation . Adoptive transfer of these hepatic CD4+ T cells to naive mice and subsequent antigenic challenge led to suppression of T cell proliferation as well as IgG Ab responses to OVA; this effect was mostly abrogated by a blocking Ab to FasL . i.p . administration of an Ag at a high dose also generated hepatic CD4+FasL+ T cells with similar cytokine profile as T cells activated by oral administration of Ags at a high dose . Finally, we did not see an increase in FasL+ cells in the hepatic CD4+Vbeta8+ T cell subset of MRL/lpr/lpr mice given staphylococcal enterotoxin B, indicating the requirement for Fas-mediated signals . These hepatic CD4+FasL+ regulatory cells may explain the tolerogenic property of the liver and play roles in systemic hyporesponsiveness induced by an Ag administered at a high dose.

Protein Expr Purif, 2002 Mar, 24(2), 302 - 12
Production and purification of a recombinant Staphylococcal enterotoxin B vaccine candidate expressed in Escherichia coli; Coffman JD et al.; An attenuated, recombinant form of Staphylococcus enterotoxin B (rSEB) was overexpressed in Escherichia coli under transcriptional control of the T7 promoter . The 28-kDa rSEB was partially purified from soluble, intracellular protein by tangential flow filtration and differential ammonium sulfate precipitation . The intermediate product was then further purified using low-pressure liquid chromatography including hydrophobic interaction, cation exchange, and size-exclusion matrices . The final vialed product was >95% pure as determined by Coomassie blue-stained sodium dodecyl sulfate-polyacrylamide gel electrophoresis, high-pressure size-exclusion chromatography, and capillary zonal electrophoresis . The endotoxin level was <0.6 EU/mg . Final estimated yield of purified rSEB was 147 mg/L of starting culture . Purified rSEB was stable, elicited an immune response in mice, and protected mice against a lethal challenge with the native toxin .

Joint Bone Spine, 2002 Jan, 69(1), 85 - 7
Three-year outcome in a patient with Staphylococcus lugdunensis discitis; Camacho M et al.; The few reported cases of bone and joint infection by Staphylococcus lugdunensis indicate that the clinical manifestations are severe, the diagnosis elusive, and the treatment difficult . We report a case of lumbar discitis caused by Staphylococcus lugdunensis in a 67-year-old man receiving chemotherapy for stage III IgA lambda multiple myeloma . Treatment was with ofloxacin and pristinamycin for 1 year . Although he started to improve only 5 months after treatment initiation, the outcome was favorable . Follow-up at the time of this writing is 3 years.

Vojnosanit Pregl, 2001 Nov-Dec, 58(6), 689 - 94
{A severe form of falciparum malaria associated with staphylococcal endocarditis}; Mikic D et al.; A case is presented of a patient, aged 56 years, with severe form of imported malaria caused by Plasmodia falciparum . Hyperparasitemia of erythrocytes > 30% was registered, and during the course of the disease CNS dysfunction, severe anemia, acute renal failure, disseminated intravenous coagulation with manifest hemorrhagic syndrome, icterus, enterocolitis, pneumonia and staphylococcal endocarditis were developed Due to hyperparasitemia and numerous complications, antimalarial drugs such as quinidine (1,200 mg/day) and artemether (160 mg/day) were administered parenterally . Infected erythrocytes were exchanged with 2.5 litres of healthy erythrocytes suspension . Hemodialysis was also performed as well as nine-week antistaphylococcal therapy . During the treatment preparation of deplasmated blood, concentrated thrombocytes, fresh frozen plasma, cryoprecipitates, human albumins and immunoglobulins were applied, along with the correction of electrolytic dysbalance, administration of diuretic, cardiotonic, antiarrhythmic, anxiolytic, antipsychotic and antidepressive drugs . Two months after the admission the patient was released from the Clinic in good condition, with normal clinical-laboratory findings.

Clin Microbiol Infect, 1999 Apr, 5(4), 190 - 194
Failure of antibiotic therapy in Staphylococcus epidermidis infection of implantable venous access devices in patients with AIDS, as documented by molecular typing; Piketty C et al.; OBJECTIVE: To study the efficacy and safety of high-concentration antibiotic locks in association with systemic antibiotherapy in Staphylococcus epidermidis infections of totally implantable venous access devices (TIVADs) in patients with AIDS . METHODS: Thirty-one episodes of S . epidermidis TIVAD infection were observed in nine patients . Locks consisted of high concentrations of aminoglycosides or vancomycin according to antibiogram results (susceptibility results obtained with the disk diffusion technique) . Genotyping of bacterial strains was performed by pulsed-field gel electrophoresis (PFGE) in 26 of the 31 infectious episodes . RESULTS: The infections occurred within a median period of 62 catheter-days . The median duration of systemic antibiotic therapy was 17.5 days in association with a median of three antibiotic locks . Failure as defined by the occurrence of a novel episode within 2 months was observed in 17 of the 31 infections (58%) . According to the PFGE results, relapse with the same strain of S . epidermidis or reinfection with a different strain of S . epidermidis could be assigned to 10 episodes which failed to respond to therapy . Relapse was observed in six of 10 episodes . Four episodes were followed by the occurrence of a novel infection with a different S . epidermidis strain . In one patient, a relapse occurred despite TIVAD removal . CONCLUSION: Our observations further document the poor efficacy of associating antibiotic locks with systemic antibiotic therapy for the treatment of TIVAD infections in patients with AIDS.

Proc Natl Acad Sci U S A, 2002 Feb 19, 99(4), 1937 - 42
A method for optimizing potential-energy functions by a hierarchical design of the potential-energy landscape: application to the UNRES force field; Liwo A et al.; A method for optimizing potential-energy functions of proteins is proposed . The method assumes a hierarchical structure of the energy landscape, which means that the energy decreases as the number of native-like elements in a structure increases, being lowest for structures from the native family and highest for structures with no native-like element . A level of the hierarchy is defined as a family of structures with the same number of native-like elements (or degree of native likeness) . Optimization of a potential-energy function is aimed at achieving such a hierarchical structure of the energy landscape by forcing appropriate free-energy gaps between hierarchy levels to place their energies in ascending order . This procedure is different from methods developed thus far, in which the energy gap and/or the Z score between the native structure and all non-native structures are maximized, regardless of the degree of native likeness of the non-native structures . The advantage of this approach lies in reducing the number of structures with decreasing energy, which should ensure the searchability of the potential . The method was tested on two proteins, PDB ID codes and, with an off-lattice united-residue force field . For, the search of the conformational space with the use of the conformational space annealing method and the newly optimized potential-energy function found the native structure very quickly, as opposed to the potential-energy functions obtained by former optimization methods . After even incomplete optimization, the force field obtained by using located the native-like structures of two peptides, and betanova (a designed three-stranded beta-sheet peptide), as the lowest-energy conformations, whereas for the 46-residue N-terminal fragment of staphylococcal protein A, the native-like conformation was the second-lowest-energy conformation and had an energy 2 kcal/mol above that of the lowest-energy structure.

Infect Immun, 2002 Mar, 70(3), 1310 - 8
Retrieving biological activity from LukF-PV mutants combined with different S components implies compatibility between the stem domains of these staphylococcal bicomponent leucotoxins; Werner S et al.; Bicomponent leucotoxins, such as Panton-Valentine leucocidin, are composed of two classes of proteins, a class S protein such as LukS-PV, which bears the cell membrane binding function, and a class F protein such as LukF-PV, which interacts to form a bipartite hexameric pore . These leucotoxins induce cell activation, linked to a Ca(2+) influx, and pore formation as two consecutive and independently inhibitable events . Knowledge of the LukF-PV monomer structure has indicated that the stem domain is folded into three antiparallel beta-strands in the water-soluble form and has to refold into a transmembrane beta-hairpin during pore formation . To investigate the requirements for the cooperative assembly of the stems of the S and F components to produce biological activity, we introduced multiple deletions or single point mutations into the stem domains of LukF-PV and HlgB . While the binding of the mutated proteins was weakly dependent on these changes, Ca(2+) influx and pore formation were affected differently, confirming that they are independent events . Ca(2+) entry into human polymorphonuclear cells requires oligomerization and may follow the formation of a prepore . The activity of some of the LukF-PV mutants, carrying the shorter deletions, was actually improved . This demonstrated that a crucial event in the action of these toxins is the transition of the prefolded stem into the extended beta-hairpins and that this step may be facilitated by small deletions that remove some of the interactions stabilizing the folded structure.

J Natl Med Assoc, 2002 Feb, 94(2), 121 - 3
Irritant contact dermatitis complicated by deep-seated staphylococcal infection caused by a hair relaxer; Kaur BJ et al.; Chemical hair relaxers are used by many women to straighten their hair . We describe a case of a deep soft tissue staphylococcal abscess that complicated an irritant contact dermatitis from a hair relaxer treatment.

J Appl Microbiol, 2002, 92(1), 158 - 64
Monitoring of Staphylococcus xylosus DSM 20266 added as starter during fermentation and ripening of soppressata molisana, a typical Italian sausage; Di Maria S et al.; AIMS: "Soppressata molisana", a fermented sausage produced in southern Italy, is commonly obtained without starter addition . However, the use of starter cultures is more and more recommended in meat fermentation processes in order to guarantee stable production performance . In this study, the survival of the Staphylococcus xylosus DSM 20266 was evaluated during the ripening of "soppressata molisana" fermented sausage . METHODS AND RESULTS: The fastest method of RAPD-PCR was employed for discrimination of the added strain from those naturally present during the ripening of the "soppressata molisana" . The results obtained were confirmed by analysis of the DNA macrorestriction profile by PFGE . The electrophoretic pattern of bacterial total proteins was also studied, but clear differences between the different strains could not be detected . CONCLUSIONS: The RAPD technique was a valid tool for monitoring Staph . xylosus DSM 20266 in "sopressata molisana" . SIGNIFICANCE AND IMPACT OF THE STUDY: This study highlights the possibility of monitoring the presence of Staph . xylosus strains during the ripening of fermented sausages by a reliable and repeatable technique such as RAPD.

Protein Sci, 2002 Mar, 11(3), 642 - 51
Structural basis for abrogated binding between staphylococcal enterotoxin A superantigen vaccine and MHC-IIalpha; Krupka HI et al.; Staphylococcal enterotoxins (SEs) are superantigenic protein toxins responsible for a number of life-threatening diseases . The X-ray structure of a staphylococcal enterotoxin A (SEA) triple-mutant (L48R, D70R, and Y92A) vaccine reveals a cascade of structural rearrangements located in three loop regions essential for binding the alpha subunit of major histocompatibility complex class II (MHC-II) molecules . A comparison of hypothetical model complexes between SEA and the SEA triple mutant with MHC-II HLA-DR1 clearly shows disruption of key ionic and hydrophobic interactions necessary for forming the complex . Extensive dislocation of the disulfide loop in particular interferes with MHC-IIalpha binding . The triple-mutant structure provides new insights into the loss of superantigenicity and toxicity of an engineered superantigen and provides a basis for further design of enterotoxin vaccines.

Roum Arch Microbiol Immunol, 1998 Apr-Jun, 57(2), 147 - 56
Effect of viral preinfection upon cell adherence in some staphylococcus strains from specific infections or carriers; Petrasincu D et al.; The adherence of bacteria to eukaryote cells has been largely investigated as an essential step in the occurrence of bacterial infection . Some clinical and epidemiological studies have revealed the frequent association of certain viral infections with bacterial infections originating in the same ecological niche . Therefore, we investigated the effect of the viral preinfection (ADV4) of some cultivated cells (HEp-2 and IC.SK-27) upon the adherence of staphylococcus to these cells . The analysis of cell adherence within the mentioned conditions, estimated by flow cytometry, allowed of the following conclusions: 1 . bacterial adherence to cultivated and virally preinfected cells is augmented by the viral preinfection, and its value on a given cell substrate may characterize a bacterial strain; 2 . bacterial adherence to the investigated cell substrates does not correlate with the origin of the tested staphylococcus strains (infections or carriers) and some cell lines can differentiate bacterial strains depending upon the ecological niche or inside it.

Cancer Immunol Immunother, 2002 Mar, 51(1), 33 - 44 Epub 2002 Jan 11.
A mutated superantigen SEA D227A fusion diabody specific to MUC1 and CD3 in targeted cancer immunotherapy for bile duct carcinoma; Takemura S et al.; In cancer immunotherapy research, many bispecific antibodies (BsAbs) have been developed for directing T cells toward tumor cells . Recent advances in genetic engineering have made it possible to prepare immunoglobulin fragments consisting of variable domains using bacterial expression systems . Therefore, recombinant BsAbs, termed diabodies, have attracted particular attention . We have previously produced an anti-MUC1 x anti-CD3 diabody (Mx3 diabody) in an Escherichia coli ( E . coli) expression system . In order to reinforce the antitumor effects of the Mx3 diabody, mutated superantigen staphylococcal enterotoxin A (SEA) D227A was genetically fused to the Mx3 diabody . The SEA D227A fusion Mx3 diabody (SEA D227A-Mx3 diabody) thus constructed showed remarkable MUC1-specific antitumor effects when used with effector cells (lymphokine-activated killer cells with T-cell phenotype {T-LAK} and peripheral blood mononuclear cells {PBMCs}) . In the bile duct carcinoma (BDC)-xenografted severe combined immunodeficient (SCID) mouse model, it also demonstrated strong antitumor activity when administered i.v . together with T-LAK cells and interleukin-2 (IL-2) . In this experiment, the complete disappearance of tumors was observed in 3 out of 6 mice, and the other 3 showed marked retardation of tumor growth . Therefore, the SEA D227A-Mx3 diabody is considered to be a promising reagent in specific targeted immunotherapy for BDC and other MUC1-positive carcinomas . This is the first report on a diabody that is effective in treating human solid cancers in the xenografted SCID mouse experimental model.

J Heart Valve Dis, 2002 Jan, 11(1), 121 - 6; discussion 27
The CarboMedics prosthetic heart valve: experience with 1,084 implants; Santini F et al.; BACKGROUND AND AIM OF THE STUDY: The study aim was to evaluate our clinical experience with the CarboMedics heart valve prosthesis . METHODS: Between October 1991 and December 2000, 942 consecutive patients (514 males, 428 females; mean age 58+/-11 years; range: 6-78 years) underwent mechanical valve implantation with the CarboMedics prosthesis . Preoperatively, 47% of patients were in NYHA class III and 22% in class IV; in addition, 134 patients (14.2%) had undergone a previous cardiac operation . Aortic valve replacement (AVR) was performed in 469 patients (49.8%), mitral valve replacement (MVR) in 330 (35.0%), double valve replacement (DVR) in 142 (15.1%), and isolated tricuspid valve replacement (TVR) in one patient . Eighty-eight patients (9.3%) underwent associated myocardial revascularization . Mean cardiopulmonary bypass and aortic cross-clamp times for the entire group were 107+/-39 min and 74+/-24 min, respectively . RESULTS: Overall early mortality was 2.3% (6/469 AVR, 1.2%; 12/330 MVR, 3.6%; 4/142 DVR, 2.8%) . Late mortality was 3.1% (n = 29; including 17 cardiac deaths (10 were valve-related) . Mean follow up was 66+/-31 months (range: 1-109 months), and was 98% complete yielding a total follow up of 4959 years . Actuarial survival at five years for the entire group was 89.3+/-1.6% (AVR 91.1%, MVR 86.4%, DVR 90.5%) . Thromboembolism occurred in 26 patients (2.8%, 0.52%/pt-year) and major hemorrhagic events in 20 (2.1%, 0.4%/pt-year) . Nine patients (0.9%) required a reoperation, in three cases (0.3%) after Staphylococcus epidermidis-mediated endocarditis . No structural deterioration occurred . Among 891 survivors, 94% of the patients are currently in NYHA classes I or II (p <0.05) . CONCLUSION: This study confirmed the safety and reliability of the CarboMedics mechanical valve prosthesis, even in old age groups . This bileaflet prosthesis showed no structural deterioration, and a low incidence of overall complications.

Immunol Lett, 2002 Apr 1, 81(1), 77 - 85
In vivo cytokine responses in gut-associated lymphoid tissue (GALT) and spleen following oral administration of staphylococcal enterotoxin B; Nishimura M et al.; The mechanisms responsible for oral tolerance have been analyzed using various antigens . The induction of oral nonresponsiveness to low doses of Staphylococcal Enterotoxin B (SEB), in which SEB-reactive clonal anergy and deletion are involved, has also been confirmed . However, few reports have examined the cytokine milieu of the mucosal and peripheral lymphoid tissue during tolerance induction after the oral administration of SEB . In this study, to elucidate the mechanism responsible for the oral tolerance to low dose of SEB, the cytokine responses of the gut-associated lymphoid tissues (GALT) and the spleen were examined using the reverse transcription-polymerase chain reaction (RT-PCR) method . After the oral administration of a single low dose of SEB, the expressions of Th2 and TGF-beta, and to a lesser extent Th1 mRNA were observed in the mucosa . In contrast, following repeated oral low doses of SEB, in a manner different from previous results using conventional antigens, no cytokine enhancements were demonstrated in the GALT or the spleen . In conclusion, the involvement of unknown inhibitory or regulatory cytokines, together with SEB-reactive clonal anergy and deletion, is suggested in the induction of oral tolerance to low dose SEB.

Pharmacotherapy, 2002 Feb, 22(2 Pt 2), 45S - 54S
Approaches to analysis of length of hospital stay related to antibiotic therapy in a randomized clinical trial: linezolid versus vancomycin for treatment of known or suspected methicillin-resistant Staphylococcus species infections; Li JZ et al.; As length of hospital stay (LOS) represents about 70-90% of the total cost of treating serious infections, it represents a key variable in analyzing the health economic differences between treatments for hospitalized patients . In a retrospective analysis using LOS data from a multinational, randomized, phase III clinical trial, we examined two methods (the log-logistic model and Kaplan-Meier survival function) and three approaches (unadjusted total LOS, total LOS adjusted for nontreatment factors, and adjusted LOS based on antibiotic treatment {the antibiotic treatment LOS}) for estimating antibiotic treatment effect on LOS and determined if these approaches could reduce the variation in LOS and control for the imbalance between treatment groups . The trial enrolled patients who were hospitalized with known or suspected Staphylococcus species infections who received at least one dose of linezolid or vancomycin (intent-to-treat sample) and who continued taking the study drug for at least 7 days (clinically evaluable sample) . In the intent-to-treat sample, the linezolid group had a 2- (unadjusted) or 4-day (adjusted for nontreatment factors) shorter LOS at the 25th percentile; a 1- or 2-day advantage, respectively, at the 50th percentile (median); and a 0.6- or 1.6-day mean LOS advantage, compared with the vancomycin group . With the antibiotic treatment LOS approach, the linezolid group had mean and median LOS reductions comparable to or greater than those seen in the nontreatment-factor-adjusted results . Results for the clinically evaluable sample were similar to those of the intent-to-treat sample, but the differences between the treatment groups were greater . Linezolid-treated patients had significant LOS reductions that otherwise would be masked without the use of more appropriate, but less commonly used, methods.

Biotechnol Appl Biochem, 2002 Feb, 35(Pt 1), 55 - 60
Sephadex-based cell-affinity adsorbents: preparation and performance; Besselink G et al.; Sephadex was derivatized consecutively with Staphylococcus Protein A (SpA) and cell-specific antibodies, and the binding of cells to the resulting material was examined . For comparison, cell binding to commercially obtained SpA-Sepharose was determined . Sephadex G-10, carboxylated by reaction with glycine and activated subsequently with 1-ethyl-3-(3-dimethylaminopropyl)carbodi-imide/N-hydroxysuccinimide (NHS), was allowed to react with SpA . Coupling of SpA to NHS-activated glycine-Sephadex appeared to be complete (immobilization capacity, approximately 300 microg of protein/ml of packed gel) when incubation was carried out at pH 4.0, in buffer of low ionic strength . However, incubation at higher pH values (> or = 6.5) led to poor coupling yields . After incubation with rabbit anti-(human red cell) antiserum, and upon mixing with human red blood cells, SpA-glycine-Sephadex G-10 could bind up to 5 x 10(8) red cells/ml of gel . Cell binding increased when the amount of antiserum, added to SpA-glycine-Sephadex G-10 for preparing the affinity gel, was increased from 0.5 to 5 microl/ml of gel . Compared with this, SpA-Sepharose CL 4B had to be incubated with much larger amounts of antiserum (100-700 microl/ml of gel) in order to obtain cell-affinity adsorbent . One obvious advantage of the approach described here is that relatively small amounts of SpA and antisera are needed for preparing cell-affinity media.

Ann Surg Oncol, 2002 Jan-Feb, 9(1), 82 - 7
Cytokine profiles of sentinel lymph nodes draining the primary melanoma; Leong SP et al.; BACKGROUND: The sentinel lymph node (SLN) draining the primary melanoma is the first echelon node where micrometastasis is established . SLNs may be the initial sites of antigen presentation associated with immune responses . METHODS: A portion of each SLN from 68 melanoma patients undergoing selective SLN dissection was processed for enzyme-linked immunospot (ELISPOT) assay determination of interferon gamma (IFN-gamma), interleukin-2 (IL-2), granulocyte macrophage colony-stimulating factor (GM-CSF), and interleukin-10 (IL-10) secretion . The control was the adjacent non-SLN . Lymphocytes were stimulated with staphylococcal enterotoxin A (SEA) prior to ELISPOT assay . RESULTS: No significant difference was noted in the production of IL-10 between the SLNs and non-SLNs . Significant production of IFN-gamma, IL-2, and GM-CSF was noted in the SLNs when compared to the non-SLNs in the overall group . Patients with no micrometastasis (n = 60) had elevated secretion of all cytokines in the SLNs . However, patients harboring lymph node micrometastasis (n = 8) showed no increase of cytokine secretion in the SLNs . CONCLUSIONS: Significant Th1 and Th2 response was induced in melanoma-free SLNs . IMPLICATIONS: SLNs without micrometastasis may be activated by submicroscopic cells or soluble tumor antigens, while cytokine production may be down-regulated by micrometastasis . Future studies should be directed towards identifying the specific SLN T cells recognizing the tumor antigens.

Cell Immunol, 2001 Nov 1, 213(2), 149 - 57
Participation of Vbeta13(+) and Vbeta1(+) T cells in transfer thyroiditis after activation of mouse thyroglobulin-primed T cells by superantigen staphylococcal enterotoxin A; Wan Q et al.; Murine experimental autoimmune thyroiditis (EAT) is a T-cell-mediated disease, but the T cell receptor (TCR) Vbeta gene usage in pathogenesis has not been well delineated . One approach is to utilize bacterial superantigens, such as staphylococcal enterotoxin (SE) A and B, to stimulate known sets of TCR Vbeta families in mouse thyroglobulin (mTg)-primed cells for thyroiditis transfer . Our previous use of SEB to activate mTg-primed cells led to no thyroiditis transfer, despite a major increase in Vbeta8(+) T cells . Unlike SEB, SEA activation did transfer thyroiditis . To determine which thyroiditogenic Vbeta(+) T cells were involved, SEA-activated T cells have now been analyzed . After repeated SEA activation in vitro, both mTg-reactive and thyroiditogenic cells persisted . FACS analysis indicated that most Vbeta13(+) cells were "large" cells (IL-2R(+)) and expressed the activation marker, transferrin receptor (CD71) . RT-PCR analysis also showed the presence of both Vbeta13(+) and SEA-reactive Vbeta1(+) cells . Since our previous analyses by RT-PCR of the thyroid infiltrate after either induction or adoptive transfer have implicated both Vbeta13(+) and Vbeta1(+) cells, their activation by SEA to transfer thyroiditis further supports their role.

Biomaterials, 2002 Mar, 23(6), 1495 - 502
Effects on antibiotic resistance of Staphylococcus epidermidis following adhesion to polymethylmethacrylate and to silicone surfaces; Arciola CR et al.; A number of studies appears to give emphasis to the role of prosthetic materials in determining microbial adherence and resistance to host defence and drug therapy . Aim of this study was to explore whether the direct contact with biomaterial substrata of different chemical nature could influence bacterial behaviour, determining possible changes in the bacteria population as far as antibiotic resistance is concerned . To this end, susceptibility to penicillin, erythromycin, clindamycin, cefamandole, imipenem, vancomycin, ciprofloxacin . ampicillin, cefazolin, trimethoprim-sulfamethoxazole, chloramphenicol, amikacin and netilmicin was evaluated in a methicillin-, gentamicin- and tobramycin-resistant Staphylococcus epidermidis strain, after in vitro adhesion to polymethylmethacrylate (PM MA) and to silicone elastomer . The susceptibility to antibiotics of both adherent bacteria and bacteria which, although exposed to the materials, had not undergone adhesion was measured as bacterial growth inhibition area onto a plate antibiogram . according to Kirby-Bauer and using an image analyser system . The results obtained suggest that the two test materials considered in this study were capable to condition bacterial behaviour . In particular . the adhesion onto PMMA surfaces induced a marked and significant decrease in susceptibility to the following beta-lactam antibiotics: cefamandole (32%), cefazolin (23%), imipenem (27%), ampicillin (31%) . Moreover, PMMA caused a lower but significant reduction in resistance to vancomycin (15%), chloramphenicol (16%), amikacin (13%) . netilmicin (13%), erythromycin (11%) trimethoprim-sulfamethoxazole (13%) . In contrast, the adhesion onto silicone elastomer appeared to influence bacterial changes to a lesser extent and elicited a significant decrease in susceptibility only to cefazolin (10%) and amikacin (11) . Further studies are required to thoroughly investigate the mechanisms of these variations, even though, also according to other authors, one of the best conceivable conclusions is that some material substrata can lead to selection of variant adhesive bacteria with increased antibiotic resistance.

J Chromatogr A, 2002 Jan 4, 942(1-2), 157 - 66
Strategy for highly selective ion-exchange capture using a charge-polarizing fusion partner; Graslund T et al.; To achieve efficient recovery of recombinantly produced target proteins using cation-exchange chromatography, a novel basic protein domain is used as a purification handle . The proteolytic instability usually encountered for basic peptide tags is avoided by the use of a highly constrained alpha-helical domain based on staphylococcal protein A into which positively charged amino acids have been introduced . Here we show that this domain, consisting of 58 amino acids with a calculated isoelectric point (pI) of 10.5, can be used to efficiently capture different fused target proteins, such as a bacterial DNA polymerase (Klenow fragment), a viral protease (3C) and a fungal lipase (Cutinase) . In contrast to standard cation-exchange chromatography, efficient capture can be achieved also at a pH value higher than the pI of the fusion protein, demonstrated here by Zbasic-Klenow polymerase (pI approximately/= 5.8) and ZZ-Cutinase-Zbasic (pI approximately/= 7.2) both purified at a pH of 7.5 . These results show that the Zbasic domain is able to confer a regional concentration of positive charge on the fusion protein even at a relatively high pH . Hence, the data suggest that this domain could be used for highly efficient and selective capture of target proteins at conditions where most host-cell proteins do not bind to the chromatographic resin . The obtained purity after this one-step procedure suggests that the strategy could be an alternative to standard affinity chromatography . Methods for site-specific proteolysis of the fusion proteins to release native target proteins are also discussed.

World J Gastroenterol, 2001 Apr, 7(2), 216 - 21
Preparation and activity of conjugate of monoclonal antibody HAb18 against hepatoma F(ab')(2) fragment and staphylococcal enterotoxin A; Yang LJ et al.; AIM: To prepare the conjugate of staphylococcal enterotoxin A (SEA) protein which is a bacterial SAg and the F(ab')(2) fragment of mAb HAb18 against human hepatocellular carcinoma (HCC), and identify its activity in order to use SAg in the targeting therapy of HCC . METHODS: MAb HAb18 was extracted from the abdominal dropsy of Balb/c mice, and was purified through chromatography column SP 40HR with Fast protein liquid chromatography (FPLC) system . The F(ab')(2) fragment of mAb HAb18 was prepared by papainic digestion method . The conjugate of mAb HAb18 F(ab')(2) fragment and SEA was prepared with chemical conjugating reagent N succinimidyl 3 (2-pyridyldithio) propionate (SPDP) and purified through chromatography column Superose 12 with FPLC system . The molecular mass and purity of each collected peak were identified with SDS-PAGE assay . The protein content was assayed by Lowry's method . The antibody activity of HAb18 F(ab')(2) against HCC in the conjugate was identified by indirect immunocytochemical ABC method, and the activity of SEA in the conjugate to activate peripheral blood mononuclear cells (PBMC) was identified with MTT assay . RESULTS: The IgG mAb HAb18 was extracted, and purified successfully . Immunocytochemical staining demonstrated that it reacted with most of HHCC cells of human HCC cell line . There were two peaks in the process of purification of the prepared HAb18 F(ab')(2) SEA conjugate . SDS-PAGE assay demonstrated that the molecular mass of the first peak was about 130 ku, and the second peak was the mixture of about 45 ku and a little 100 ku proteins . The immunocytochemical staining was similar in HAb18 F(ab') (2) SEA conjugate and HAb18 F(ab') (2), i.e.the cytoplasm and/or cell membranes of most HHCC cells were positively stained . The MTT assay showed that the optical absorbance (A) value at 490 nm of HAb18 F(ab') (2) SEA conjugate was 0.182 +/- 0.012, that of negative control was 0.033 +/- 0.009, and there was significant difference between them (P < 0.05) . CONCLUSION: SPDP is a good protein conjugating reagent and can be used in preparing protein conjugate . The conjugate of mAb HAb18 F(ab') (2) fragment and SEA protein was prepared successfully in present study and can be used in the experimental study of HCC targeting therapy with the conjugate of SAg and anti HCC mAbs or their fragments.

World J Gastroenterol, 1999 Jun, 5(3), 209 - 212
Cyclosporin A protects Balb/c mice from liver damage induced by superan tigen SEB and D-GalN; Yin T et al.; AIM:To investigate the pathogenic effect of SEB and D-GalN on liver and the protection of cyclosporin A, the relationship between hepatic apoptosis and necrosis and the possible mechanism of acute hepatic necrosis.METHODS:After staphylococcal enterotoxin B (SEB) mixed with D-galactosamine (D-GalN) were injected intraperitoneally into Balb/c mice and those previously treated with cyclosporin A, blood samples were collected and livers were isolated at 2, 6, 12, 24h . Patterns of hepatocellular death were studied morphologically and biochemically, circulating cytokines (TNF-alpha, IFN-gamma) and mice mortality within 24h was assessed.RESULTS: The SEB could induce the typical apoptotic changes of hepatocytes, the D-GalN could induce hepatocytes apoptosis and degeneration at the same time, and the mice having received the SEB+D-GalN injections developed apoptosis at 2 and 6h, but after 12h hepatocytes were characterized by severe injury, whereas all the examinations in the cyclosporin A treated mice were normal.CONCLUSION:Hepatic cell apoptosis might be related to necrosis, and massive hepatocyte apoptosis is likely the initiating step of acute hepatic necrosis in mice . The effects induced by SEB and D-GalN on hepatocytes might be mediated by T cells, and could be prevented by cyclosporin A.

Vaccine, 2002 Jan 31, 20(9-10), 1354 - 64
Double mutant and formaldehyde inactivated TSST-1 as vaccine candidates for TSST-1-induced toxic shock syndrome; Gampfer J et al.; Up to now there is no treatment for staphylococcal toxic shock syndrome, a disease mainly induced by toxic shock syndrome toxin-1(TSST-1) . There is great demand in finding means to control the disease, one of them is the development of an effective and safe vaccine against TSST-1 . In this study we constructed a series of vaccine candidates and investigated their biological activity, toxicity, and potential to invoke an immune response . TSST-1 was isolated from Stahylococcus aureus supernatants and recombinantly expressed as a N-terminal 6x histidine-tagged protein in Escherichia coli . In order to obtain molecules with minimal toxicity we constructed single mutants (G31R and H135A) and one double mutant (G31R/H135A) with both residues exchanged . We also detoxified native TSST-1 isolated from S . aureus, and recombinantly expressed TSST-1 by treatment with formaldehyde . Functional activity of native and recombinant TSST-1 and grade of inocuity of mutants and toxoids was determined by investigating mitogenity, T-cell activation, and cytokine release upon stimulation of human mononuclear cells with the vaccine candidates . All substances were tested in a rabbit immunization study . After primary immunization and three additional boosts all vaccinated animals developed antibody titers against TSST-1 and were protected against challenge with a lethal doses of superantigen potentiated with lipopolysaccharide.

J Endocrinol Invest, 2001 Dec, 24(11), 887 - 91
Magnetic resonance imaging and computerized tomography images in a case of pituitary abscess; Erdogan G et al.; A 29-year-old male patient with clinical manifestations of panhypopituitarism and diabetes insipidus is presented . The clinical and laboratory evaluation of the pituitary reserve confirmed a total pituitary insufficiency . Computed tomography (CT) scan demonstrated a low-density lesion suggesting a pituitary tumor with suprasellar extension, and magnetic resonance imaging (MRI) revealed a pituitary mass with decreased signal intensity on T1-weighted images and capsular contrast enhancement after gadolinium injection . The surgical procedure was the treatment of choice . During surgery a large amount of purulent material was removed . With light microscopy, chronic non-specific inflammation and a pituitary abscess capsule were demonstrated in the tissue . The results of the cultures revealed coagulase negative staphylococcus . The post-operative course was uneventful and, as the patient was pituitary-deficient, he was on replacement therapy and was being followed-up . Pituitary abscess is an exceptional lesion in the literature and despite the advent of CT and MRI, its preoperative diagnosis still remains difficult . However, the presence of an intrasellar expansive process with liquid center and contrast enhanced outline should suggest the possibility of an abscess . Sellar round cystic mass isointense or hypointense to grey matter on T1, high intensity signal on T2, or opposite pattern with a peripheral rim enhancement following gadolinium injection, and diabetes insipidus may all be suggestive of a pituitary abscess.

Int J Artif Organs, 2001 Oct, 24(10), 671 - 82
Prevention of catheter related infections in patients on CAPD; Thodis E et al.; Catheter-related infections remain a serious problem for patients on peritoneal dialysis . Such infections can be reduced by careful patient selection and training, by the use of the best connection technology and screening and treating nasal carriage . To date, treatment is less than optimal and therefore, the primary goal should be prevention of catheter-related infections . Prevention is based on improving catheter design and implantation technique, while providing careful exit-site care . Regardless of how it is implemented, we must aggressively pursue the prevention of catheter-related infections by eradicating S . aureus exit-site carriage in PD patients . Based on its effectiveness in adult PD patients, its low rate of adverse effects, and its reasonable cost-effectiveness, application of mupirocin ointment at the exit-site is the current method of choice for preventing PD catheter infections caused by S . aureus . In addition to reducing S . aureus exit-site infections, mupirocin seems to reduce the rates of staphylococcal peritonitis and PD catheter loss . Whether the ointment should be applied in the nares, to the exit-site or both, and whether it should be used only in staphylococcal nasal carriers or all PD patients requires further study.

Biochem Biophys Res Commun, 2002 Feb 1, 290(4), 1336 - 42
Superantigen-induced apoptotic death of tumor cells is mediated by cytotoxic lymphocytes, cytokines, and nitric oxide; Mondal TK et al.; The bacterial superantigen staphylococcal enterotoxin A (SEA) is a potent inducer of CTL activity and cytokine production in vivo . Protein A (PA) of Staphylococcal aureus has been found to have diverse biological response modifying properties and to possess antitumor, antitoxic and antiparasitic effects . In this study we examined the anti-tumor effect of these two superantigens used separately as well as in combination in mice carrying the Ehrlich ascites tumor . With combined treatment, DNA cell cycle analysis of tumor cells showed a significant (P < 0.05) percentage of tumor cell death . Levels of the soluble mediators TNF-alpha, IFN-gamma and IL-1 as well as NO were elevated . Additionally, CD4(+) and CD8(+) specific T cells in spleen, thymus and PBMC in tumor carrying mice were increased (P < 0.01) . Our data altogether suggests that enhanced tumor cell death is caused by the increased CTL activity, cytokine and nitric oxide levels, in response to the combined effect of SEA + PA . (c)2002 Elsevier Science (USA).

J Infect Chemother, 2001 Sep, 7(3), 163 - 8
Epidemiological study of vancomycin-resistant enterococcus isolated from a single medical university hospital in Japan; Ishikawa K et al.; Since 1998 more than 50 reports have described the isolation of high-level vancomycin-resistant enterococci (VRE) in Japan . Here, we report on our clinical isolates of VRE and an epidemiological study carried out using chemical and genetic techniques . VRE isolates were screened for high resistance to vancomycin (VCM) with a cutoff value of 6 microg/ml and VCM-resistant gene was confirmed by polymerase chain reaction (PCR) . The epidemiological studies used pulsed-field gel electrophoresis (PFGE) and plasmid analysis . Six strains of VRE were isolated from six different patients on two wards during a 3-months period . All of the isolates possessed vanA on their plasmid, and the isolates were divided into two similar groups . Furthermore, three different patterns were defined by PFGE . Although all of the asymptomatic carriers were hospitalized for more than 3 months, we were able to prevent an outbreak of VRE in our hospital by using our guidelines for infection control, which are stricter than those for methicillin-resistant Staphylococcus aurens . From the results of this epidemiological study, we propose that there was a possibility of contamination in this hospital, and that three of the six isolates may have acquired vanA independently . In this study, we demonstrated that infection control, according to appropriate prevention guidelines, as well as regular surveillance for VRE, are essential for designing interventions to prevent the further spread of VRE.

J Infect Chemother, 2001 Sep, 7(3), 142 - 7
Glycopeptide susceptibility profiles of nosocomial multiresistant Staphylococcus haemolyticus isolates; Tabe Y et al.; We investigated 48 Staphylococcus haemolyticus isolates from patients and medical staff in terms of susceptibility to and in-vitro selection for vancomycin and teicoplanin in regard to their antibiotypes . On comparison of multiresistant S . haemolyticus isolates with non-multiresistant isolates, the geometric mean minimum inhibitory concentration (MIC) of vancomycin for multiresistant S . haemolyticus was 2.9 microg/ml, and that of teicoplanin was 18.0 microg/ml, both of which values were significantly greater than the corresponding mean MICs of vancomycin (2.0 microg/ml) and teicoplanin (4.7 microg/ml) for nonmultiresistant isolates . After agar selection, the mean of the highest teicoplanin concentration of selected plates for multiresistant S . haemolyticus was 97.1 microg/ml, which was significantly higher than that for nonmultiresistant isolates (57.8 microg/ml) . However, the means' of the highest vancomycin concentrations after agar selection for multiresistant and nonmulti-resistant isolates were the same, at 7.4 microg/ml, with no colonies capable of growing in 32 microg/ml of vancomycin . There was no significant difference in glycopeptide susceptibility between oxacillin-resistant and oxacillin-susceptible isolates among nonmultiresistant S . haemolyticus . The geometric mean MICs of vancomycin for oxacillin-resistant and oxacillin-susceptible isolates were 2.1 microg/ml and 1.6 microg/ml, and those of teicoplanin were 4.4 microg/ml and 5.6 microg/ml, while the means of the highest concentrations of the selected plates of vancomycin were 8.6 microg/ml and 3.3 microg/ml, and those of teicoplanin were 52.8 microg/ml and 74.7 microg/ml, respectively . Multiresistant isolates showed significantly greater mean MICs of vancomycin and teicoplanin and higher teicoplanin concentration of the selected plates than nonmultiresistant isolates, irrespective of oxacillin resistance . These results indicate that methicillin resistance may not be related to reduced susceptibility to glycopeptide in S . haemolyticus, and that a multiresistant profile is associated more with a decreasing susceptibility to glycopeptides then with resistance to oxacillin . In this study, antibiotypes showed good concordance with pulsed-field gel electrophoresis typing results, with a sufficiently high discriminatory ability index, of 0.912 . We consider that primary screening with antimicrobial susceptibility testing and antibiotyping, with attention to the multiresistant profile, would be useful for monitoring nosocomial S . haemolyticus colonization and infection.

Biotechniques, 2002 Jan, 32(1), 178 - 80, 182-3
Use of magnetic beads in selection and detection of biotoxin aptamers by electrochemiluminescence and enzymatic methods; Bruno JG et al.; Systematic evolution of ligands by exponential enrichment (SELEX) was used to develop DNA ligands (aptamers) to cholera whole toxin and staphylococcal enterotoxin B (SEB) . Affinity selection of aptamers was accomplished by conjugating the biotoxins to tosyl-activated magnetic beads . The use of magnetic beads reduces the volumes needed to perform aptamer selection, thus obviating alcohol precipitation and allowing direct PCR amplification from the bead surface . Following five rounds of SELEX, 5'-biotinylated aptamers were bound to streptavidin-coated magnetic beads and used for the detection of ruthenium trisbypyridine {Ru(bpy)3(2+)}-labeled cholera toxin and SEB by an electrochemiluminescence methodology . A comparison of control (double-stranded) aptamer binding was made with aptamers that were heat denatured at 96 degrees C (single-stranded) and allowed to cool (conform) in the presence of biotoxin-conjugated magnetic beads . Results suggest that control aptamers performed equally well when compared to heat-denatured DNA aptamers in the cholera toxin electrochemiluminescence assay and a colorimetric microplate assay employing peroxidase-labeled cholera toxin and 5'-amino terminated aptamers conjugated to N-oxysuccinimide-activated microtiter wells . Interestingly, however, in the SEB electrochemiluminescence assay, double-stranded aptamers exceeded the performance of single-stranded aptamers . The detection limits of all aptamer assays were in the low nanogram to low picogram ranges.

Bone Marrow Transplant, 2001 Dec, 28(12), 1105 - 9
Immunologic recovery after autologous blood stem cell transplantation in patients with AL-amyloidosis; Akpek G et al.; We prospectively studied absolute lymphocyte (ALC) and monocyte counts (AMC), lymphocyte subsets and proliferative in vitro responses to mitogen and antigen in 12 patients with AL-amyloidosis (AL) undergoing autologous blood stem cell transplantation (SCT) with high-dose i.v . melphalan . Myeloid and lymphoid recovery (>500 per microl) occurred in a median of 10 days post SCT . While there was a continuous decline in the number of CD4+ T cells at 3 months, ALC, AMC, B cells (CD19+), CD8+ T cells, and NK cells (CD16+/56+) returned to baseline . While T cell proliferative responses to phytohemagglutinin (PHA) remained depressed, B cell function measured by the proliferative response to staphylococcal antigen returned to baseline by 3 months . To supplement our findings, we retrospectively evaluated ALC, AMC and serum immunoglobulin levels in a separate group of patients treated with the same protocol at our institution . ALC and AMC recovery was similar to the pattern observed in the initial study group . Immunoglobulin levels remained within normal ranges at 3 and 12 months after SCT . Of 50 patients who were followed for a minimum of 1 year following SCT, seven (14%) developed shingles and one (2%) had PCP pneumonia . In conclusion, cellular immune function, reflected by absolute numbers of CD4+ T cells and PHA responsive T cell proliferation, is significantly suppressed at 3 months after SCT in patients with AL, and this post-transplant immunosuppression is associated with a low but clinically meaningful occurrence of opportunistic infections typical of T cell immunosuppression.

Phys Rev E Stat Nonlin Soft Matter Phys . 2002 Jan;65(1 Pt 1):011901 . Epub 2001 Dec 11.
Model for calculation of electrostatic interactions in unfolded proteins; Kundrotas PJ et al.; An approach for the calculation of electrostatic interactions and titration properties of unfolded polypeptide chains (denatured proteins) is proposed . It is based on a simple representation of the denatured proteins as a state with titratable sites distributed on the surface of a sphere, radius of which is assumed to be equal to the radius of gyration, R(g), of an unfolded molecule . Distances between the charges, d, obey constraints arising from the protein sequence . Criteria for evaluation of the parameters R(g) and d were obtained from computer simulations on a polypeptide consisting of 20 identical amino acids (polylysine) . The model was applied for calculation of titration curves of denatured barnase and staphylococcal nuclease . It was demonstrated that the approach proposed gives considerably better agreement with the experimental data than the commonly used null approximation . It was also found that titration properties of denatured proteins are slightly, but distinguishably influenced by the amino-acid sequence of the protein.

Antimicrob Agents Chemother, 2002 Feb, 46(2), 543 - 5
Antimicrobial activity of prosthetic heart valve sewing cuffs coated with minocycline and rifampin; Darouiche RO et al.; Prosthetic heart valve sewing cuffs coated with minocycline and rifampin exhibited in vitro zones of inhibition against all 52 tested clinical isolates responsible for prosthetic valve endocarditis . An in vitro elution study of these coated sewing cuffs demonstrated residual zones of inhibition against Staphylococcus epidermidis for at least 4 weeks.

Antimicrob Agents Chemother, 2002 Feb, 46(2), 327 - 32
Efficacy of minocycline and EDTA lock solution in preventing catheter-related bacteremia, septic phlebitis, and endocarditis in rabbits; Raad I et al.; To determine the efficacy of antibiotic catheter lock solution in preventing catheter-related infections, silicone catheters were tunneled and inserted into the jugular veins of 18 rabbits . The catheters were challenged with an intraluminal injection of 10(5) CFU of slime-producing Staphylococcus epidermidis in 0.1 ml of water . The catheters were maintained on heparin (100 IU/ml) flush for the first 3 days . On day 3, quantitative blood samples for culture were obtained from the catheters and ear veins, which documented catheter-related bacteremia, and the rabbits were randomized to have their catheters flushed as follows: five animals were continued on heparin (100 IU/ml), five animals received vancomycin (3 mg/ml) with heparin (100 IU/ml), and eight animals received 3 mg of minocycline per ml with 30 mg of EDTA per ml (M-EDTA) . All animals were killed at day 7 . Blood, catheters, jugular veins, and heart valves were cultured quantitatively . Animals maintained on heparin developed catheter-related colonization, bacteremia, septic phlebitis, and endocarditis . Vancomycin-heparin partially prevented catheter colonization, bacteremia, and phlebitis (P = 0.2) . M-EDTA completely prevented catheter colonization, catheter-related bacteremia, and phlebitis in all of the animals (P < 0.01) . Tricuspid endocarditis was equally prevented by vancomycin-heparin and M-EDTA (P < or = 0.06) . In conclusion, the M-EDTA catheter flush solution was highly efficacious in preventing catheter-related colonization, bacteremia, septic phlebitis, and endocarditis in rabbits.

Pediatr Infect Dis J, 2002 Jan, 21(1), 22 - 7
Central venous catheter removal versus in situ treatment in neonates with coagulase-negative staphylococcal bacteremia; Karlowicz MG et al.; OBJECTIVE: To determine how often neonates with coagulase-negative staphylococcal (CONS) bacteremia can be treated successfully without removing the central venous catheter (CVC) . METHODS: A cohort study of CONS bacteremia and CVCs was conducted in infants in a neonatal intensive care unit in a 5-year period (1994 through 1998) . CONS bacteremia was defined as at least two positive blood cultures within 3 days of each other . RESULTS: Fifty-six infants had early removal CVC (ER-CVC) within 3 days, and 63 infants had late removal CVC (LR-CVC) >3 days after the first positive blood culture . All cases of CONS bacteremia were treated with vancomycin . There was no significant difference between infants in the ER-CVC and LR-CVC groups in terms of recurrence of bacteremia or case fatalities . CONS bacteremia of >3 days duration was more frequent in LR-CVC patients than ER-CVC patients: 43% vs . 13% (relative risk, 3.4; 95% confidence interval, 1.6 to 7.2) . CONS bacteremia was successfully treated without CVC removal in 46% of LR-CVC cases . Seventy-nine percent of LR-CVC cases with CONS bacteremia lasting 1 or 2 days were treated successfully without CVC removal . The success rate decreased to 44% with a 3- to 4-day duration of bacteremia . None of 19 infants with CONS bacteremia lasting >4 days was treated successfully until CVCs were removed . CONCLUSIONS: Prolonged CONS bacteremia was avoided by early removal of CVCs . Retention of CVCs was successful in 46% of neonates with CONS bacteremia in whom it was attempted, but it was never successful if bacteremia lasted >4 days.

Rev Med Liege, 2001 Nov, 56(11), 745 - 7
{Clinical case of the month . Acute staphylococcal scalded skin syndrome in a fifty-year-old}; Fumal I et al.; Staphylococcal scalded skin syndrome is a bullous disease induced by bacterial exfoliative exotoxins . Children are most often affected . The prognosis is worse in adults and calls for treatment in an intensive care unit . We report a case in a quinquagenerian woman who suffered from angina a few days before the bullous erythroderma.

Hepatology, 2002 Jan, 35(1), 190 - 8
Frequencies of HCV-specific effector CD4+ T cells by flow cytometry: correlation with clinical disease stages; Rosen HR et al.; Hepatitis C virus (HCV) is the leading cause of chronic hepatitis, affecting approximately 2% of the world's population . The immune mechanisms responsible for the highly variable natural history in a given individual are unknown . We used a multiparameter flow cytometric technique to functionally and phenotypically characterize HCV-specific effector T cells in the peripheral blood of 32 individuals with different stages of hepatitis C disease (resolved, mild chronic, advanced chronic) and normal controls . We found the highest frequencies of virus-specific effector cells with an activated memory phenotype (CD45RO+CD69+) in subjects who had resolved HCV infection, either spontaneously or with antiviral therapy . Effector cells from patients with resolved infection produced Th1 type cytokines following stimulation with nonstructural antigens (NS3 and NS4), whereas effector cells from chronically infected patients produced Th1 type cytokines predominantly following stimulation with the HCV core antigen . Stimulation with superantigen staphylococcal enterotoxin (SEB) induced the same levels of cytokine production in the different patient groups . Among the HCV-seropositive patients, viral load inversely correlated with the Th1 effector cell response to NS3 . Interleukin (IL)-4 was produced only in response to the control antigens, but not in response to the HCV recombinant proteins . Taken together, these findings suggest that a vigorous HCV-specific CD4+ Th1 response, particularly against the nonstructural proteins of the virus, may be associated with viral clearance and protection from disease progression . Prospective studies using this new flow cytometric assay will be required to determine whether antiviral therapy modifies the frequency, specificity, and function of these virus-specific effector cells.

Zhonghua Gan Zang Bing Za Zhi, 2001 Dec, 9(6), 343 - 5
{Influence of staphylococcal enterotoxin A on immunogenicity of hepatocellular carcinoma cells}; Li Z et al.; OBJECTIVE: To transduce the gene of staphylococcal enterotoxin A (SEA) into a HCC cell line HHCC and to make the cells to express the molecules of SEA for the purpose of improving the immunogenicity and inducing immune rejection of HCC . METHODS: Construct retroviral vector containing the gene of SEA and transduce it into HHCC cells . Then the integration, transcription and expression of recombinant were analyzed . At last cytotoxicity test of peripheral blood mixed lymphocytes was carried out . RESULTS: We obtained the retroviral vector pLXSN-SEA, transduced into HHCC cells and obtained the HCC cell line that expressed SEA toxin protein . The content of SEA in supernatant was at the level of pg . The cytotoxicity of HHCC transduced with SEA by T lymphocytes was 45.6%, which was higher than that of HHCC (20.7%) . The Km value of T lymphocytes against HHCC transduced with SEA and without SEA was 5.18 x 10(4) and 2.92 x 10(5), separately . The difference had statistical significance . CONCLUSIONS: After transducing the SEA gene, although tiny quantity of expression is detected, the cytotoxicity assay proves that the expression of SEA lead to a robust immune response.

Rev Hosp Clin Fac Med Sao Paulo, 2001 Sep-Oct, 56(5), 149 - 52
Vancomycin monitoring in term newborns: comparison of peak and trough serum concentrations determined by high performance liquid chromatography and fluorescence polarization immunoassay; Feferbaum R et al.; INTRODUCTION: Peak and trough serum concentrations of vancomycin were determined in term newborn infants with confirmed or suspected Staphylococcus sp sepsis by high performance liquid chromatography and flourescence polarization immunoassay . OBJECTIVE: To statistically compare the results of the high performance liquid chromatography and flourescence polarization immunoassay techniques for measuring serum vancomycin concentrations . METHODS: Eighteen peak and 20 trough serum samples were assayed for vancomycin concentrations using high performance liquid chromatography and flourescence polarization immunoassay from October 1995 to October 1997 . RESULTS: The linear correlation coefficients for high performance liquid chromatography and flourescence polarization immunoassay were 0.27 (peak, P = 0.110) and 0.26 (trough, P = 0.1045) respectively, which were not statistically significant . CONCLUSION: There was wide variation in serum vancomycin concentrations determined by high performance liquid chromatography as compared with those determined by flourescence polarization immunoassay . There was no recognizable pattern in the variability; in an apparently random fashion, the high performance liquid chromatography measurement was sometimes substantially higher than the flourescence polarization immunoassay measurement, and at other times it was substantially lower.

Clin Experiment Ophthalmol, 2001 Dec, 29(6), 400 - 5
Endophthalmitis in the western Sydney region: a case-control study; Lertsumitkul S et al.; BACKGROUND: A retrospective case-control study was conducted to investigate risk factors for endophthalmitis following routine intraocular surgery . METHODS: A review was performed of consecutive cases of endophthalmitis from three teaching hospitals in the western Sydney region and matched controls from the same institutions between 1996 and 1998 . RESULTS: There were 31 cases and 66 controls . Eighty procedures were phacoemulsification, 15 conventional extracapsular cataract extraction, and two were penetrating keratoplasties . Of the 80 patients who had phacoemulsification surgery, 50 had a clear corneal incision, and 26 had a scleral incision (four were unknown) . Logistic regression showed an increased risk of endophthalmitis with surgical complications (P = 0.002) and clear cornea temporal incisions (P = 0.007) . Risk of endophthalmitis was reduced with use of subconjunctival injections (P = 0.008) . The yield for the Gram stain was 47% and for culture was 67% . Anterior chamber tap in addition to vitreous biopsy alone did not increase the yield for microorganism (P = 0.78) . Mean visual acuity on presentation was hand movement with 13 patients (50%) showing visual improvement following intravitreal injections of antibiotics (P = 0.003) . Visual prognosis did not correlate with presenting visual acuity but appeared to be better in those who grew Staphylococcus epidermidis or were culture negative . CONCLUSIONS: Although this study is unable to draw definite conclusions regarding risk of endophthalmitis in clear corneal temporal cataract surgery, sufficient data suggest the importance of incision type and location . Surgical complication is an important risk factor for endophthalmitis . Use of subconjunctival antibiotic injections at the conclusion of the procedure is recommended.

Clin Diagn Lab Immunol, 2002 Jan, 9(1), 126 - 31
Stimulant-dependent modulation of cytokines and chemokines by airway epithelial cells: cross talk between pulmonary epithelial and peripheral blood mononuclear cells; Krakauer T; Staphylococcal exotoxins (SE) and lipopolysaccharide (LPS) stimulate cells of the immune system to produce proinflammatory cytokines and chemokines which mediate septic shock and acute lung inflammation . A coculture of human peripheral blood mononuclear cells (PBMC) and pulmonary A549 epithelial cells was used to investigate inflammatory responses triggered by staphylococcal enterotoxin B (SEB), toxic shock syndrome toxin 1, and LPS . The levels of interleukin 1beta (IL-1beta), IL-6, gamma interferon-inducible protein 10, monocyte chemotactic protein 1 (MCP-1), macrophage inflammatory protein 1alpha, and RANTES were enhanced by 3.8-, 4.2-, 3.1-, 8.9-, 2-, and 2.9-fold, respectively, in cocultures of SEB-stimulated cells compared to in SEB-stimulated PBMC . In LPS-stimulated cocultures, only MCP-1 and RANTES levels were increased . These data suggest that the modulation of specific cytokines and chemokines is dependent on the stimulus and that there is bidirectional interaction between PBMC and lung epithelial cells to influence the immune response to these different stimuli.

Pathol Biol (Paris), 2001 Dec, 49(10), 812 - 4
Infection and/or colonization by methicillin-resistant Staphylococcus epidermidis (MRSE); Rabaud C et al.; We analyzed the parameters predictive of identification of methicillin-resistant Staphylococcus epidermidis (MRSE) in sample performed in hospitalized patients . One hundred six Staphylococcus epidermidis strains (60 MRSE and 46 MSSE) were collected . Three variables were independently linked to MRSE isolation in multivariate analysis: hospitalization during the month preceding the current admission; on-going antimicrobial therapy before sampling, and on-going infection at the time of sampling . MRSE isolation was associated with a poor vital prognosis . The air and surfaces sampling in the rooms of two patients with nasal MRSE carriage yielded the same strains as those carried by the patient, and could play a role in the epidemiological chain of hospital-acquired MRSE infections.

Cancer Immunol Immunother, 2001 Dec, 50(10), 539 - 48
Mutated SEA-D227A-conjugated antibodies greatly enhance antitumor activity against MUC1-expressing bile duct carcinoma; Kodama H et al.; For the purpose of establishing a new adoptive immunotherapy for bile duct carcinoma (BDC), we have directed our attention to superantigens (SAgs), the most potent known activators of T lymphocytes . In our previous study, staphylococcal enterotoxin A (SEA) was conjugated chemically with MUSE11 mAb, which recognizes the MUC1 cancer-associated antigen, and shown to enhance the specific cytotoxic activity of T-LAK cells against MUC1-expressing BDC cells (TFK-1) in vitro and in vivo . However, it is probable that SEA might cause side-effects because of nonspecific binding to class II positive cells . In order to overcome these, we generated mutated SEA (mSEA) by changing Asp at position 227 of native SEA to Ala, which has reduced affinity to MHC class II molecules, but retains the potential for T cell activation . When mSEA-D227A was administered to rabbits to examine effects on blood pressure, 500 times more mSEA-D227A was tolerated than native SEA . This prompted us to construct a mSEA-D227A-conjugated mAb, reactive with MUC1 . It augmented the antitumor activity of T-LAK cells significantly, and furthermore, mSEA-D227A could be conjugated to two bispecific antibodies, BsAb (anti-MUC1 x anti-CD3) and BsAb (anti-MUC1 x anti-CD28), which in combination had greater enhancing effects than mSEA-D227A-conjugated anti-MUC1 mAb, and combination of unconjugated BsAbs . These findings indicate a utility of mSEA-D227A-conjugated antibodies for targeted cancer immunotherapy.

Infect Control Hosp Epidemiol, 2001 Oct, 22(10), 618 - 23
An outbreak of coagulase-negative staphylococcal surgical-site infections following aortic valve replacement; Lark RL et al.; OBJECTIVES: To determine the cause of a coagulase-negative staphylococcal outbreak and to identify risk factors for surgical-site infections among patients following Medtronic Freestyle bioprosthesis implantation . DESIGN: Retrospective case-control study . SETTING: An 800-bed university referral center . PATIENTS: The cohort of 64 patients undergoing Freestyle valve replacement from September 1998 to December 1998 . RESULTS: Seven patients developed infection (10.9% vs 1.1% during the preceding 8 months), including two with mediastinitis and five with endocarditis . There were no statistically significant differences between cases and controls with respect to age, gender, weight, underlying illness, preoperative hospital stay, duration of surgery, time on bypass, central venous catheter duration, National Nosocomial Infection Surveillance risk index, New York Heart Association class, albumin, or antibiotic prophylaxis . However, only three cases were documented to have received vancomycin prophylaxis . Of all staff evaluated, only surgical resident A was significantly associated with infection (odds ratio, 7.68; 95% confidence interval, 1.3-44.1; P=.02) Pulsed-field gel electrophoresis patterns on Staphylococcus epidermidis isolates from four of the six cases were identical . These cases were performed on different days . Surgical resident Awas the only staff member present in the operating room for all cases caused by the epidemic strain . This S epidermidis strain, however, was not isolated from operating room staff . CONCLUSION: A surgical resident was significantly associated with infection . However, the cause of this outbreak was likely multifactorial . Changes occurring during the investigation included institution of vancomycin as routine prophylaxis and modification of surgical technique, which contributed to the resolution of the outbreak.

Ann Urol (Paris), 2001 Nov, 35(6), 329 - 34
{Treatment of primary pyogenic abscess of the psoas: retrospective study of 18 cases}; Dahami Z et al.; The primitive abcess of psoas muscle is a rare affection which involves mainly the young . The authors report 18 cases of primitive pyogenic abcess of psoas collected during a period of six years in the urology service of UHC Ibn Rochd Casablanca . The purpose of this review is to discuss the different therapeutic attitudes . The study concerned 11 men and seven women, their mean age was 35 years . The mean delay of evolution was 45 days . The physical exam found a psoitisis (five cases) and a flunk swelling (14 cases) . The ultrasound exam showed the collection which was hypoechogenic in seven cases and heterogenous in 11 cases . The CT scan showed an heterogenouse hypodense collection without enhansment in seven cases . The use of antibiotherapy alone (four cases) and percutaneous drainage (four cases) permitted the recovery just respectively in one and two cases . Fifteen patients (83%) were operated through a postero-lateral lumbotomy route without costal resection . The bacteriological exam of the pus found a Staphylococcis aureus (ten cases) and Escherichia coli (two cases) . The evolution was favorable in 14 cases; one patient was dead at fifth day post operatively by heart infarction . For the authors, the surgical drainage remains the treatment of choice.

J Biomed Mater Res, 2002 Mar 5, 59(3), 557 - 62
In catheter infections by Staphylococcus epidermidis the intercellular adhesion (ica) locus is a molecular marker of the virulent slime-producing strains; Arciola CR et al.; Recently, it has been shown that S . epidermidis includes the ica operon responsible for slime production . In the operon, coexpression of icaA and icaD genes is required for full slime synthesis . In this study, the presence of icaA and icaD genes was searched for in a collection of 100 Staphylococcus epidermidis strains from catheter-associated infections by an original PCR method . Another 51 strains of S . epidermidis isolated from the skin or mucosa of healthy volunteers (26 of which derived from the hospital staff) were also investigated . Slime-forming ability was phenotypically tested on Congo red agar plates . Sixty-one percent of the strains isolated from catheters were icaA- icaD-positive and produced slime . The results indicate that detection of ica genes by a PCR method is a useful tool for prompt identification of S . epidermidis slime-forming strains isolated from catheter-related infections . Also, three saprophytic strains from the hospital staff were positive for slime synthesis and presence of ica genes, suggesting a potential diffusion of slime-forming strains in hospital personnel .

Exp Gerontol, 2002 Jan-Mar, 37(2-3), 271 - 83
Age associated decline in CD25 and CD28 expression correlate with an increased susceptibility to CD95 mediated apoptosis in T cells; Dennett NS et al.; Immunosenescence is believed to contribute to increase susceptibility to infectious diseases and cancer in the elderly, and is caused mainly by changes in the T cell compartment . Longitudinal studies were undertaken to examine T cell surface receptor expression and apoptotic susceptibility using Staphylococcal enterotoxin B (SEB) activated human T cells as an in vitro model of an ageing T cell culture . An intracellular stain Carboxyfluorescein diacetate succinimidyl ester (CFSE) was used to assess the number of population divisions (PD) occurring in the ageing T cell culture . One major biomarker of aged T cells is a decrease in expression of CD28 and since this is an essential co-stimulatory molecule, its decreasing expression with age could compromise their activation and apoptotic capacity . Activation of T cells resulted in initial up-regulation of CD25, CD95 and CD28, although expression of CD25 and CD28 subsequently decreased with increasing PD . CD4 and CD8 T cells expressed similar CD25 profiles although CD28 expression was unique in each subset . CD4+ cells expressed the highest CD28 levels, and showed a gradual decline in expression with increasing PD, whereas CD8+ cells were low CD28 expressers, but did not appear to lose their expression as they aged . To determine T cell susceptibility to apoptosis via CD95/CD95-L interactions with increasing age, cells were challenged with CD95-L transfected CHO cells at various PD . Increased death was observed as they aged, which correlated with the decreased expression of activation markers CD25 and CD28.

Pol Merkuriusz Lek, 2001 Oct, 11(64), 362 - 9
{Sepsis--continual pathogenetic, diagnostic and therapeutic problem}; Zdziarski P et al.; Sepsis is defined as systemic inflammatory response syndrome (SIRS) caused by infection . Massive activation of humoral and cellular mechanisms of host defense is present and can lead to a multiple organ dysfunction syndrome . Sepsis is good example, that inflammation binds with considerable energetic and metabolic effort of organism i.e . demand for oxygen, protein and microelements . The effort during SIRS exceeds compensatory abilities of organism, cause shock and death . The high mortality (40-70% in the case of septic shock) is a good illustration of the therapeutic problem . In the paper pathogenesis, diagnosis and therapy of sepsis was described . Pathogenetic importance of intoxication was explained--massive intoxication (for example by Staphylococcal enterotoxin) without infection cans lead to SIRS too . Precious advice in selection of antibiotic were given . Advantage penicillins and karbapenems over cephalosporins result from binding to PBP 2 or 1, considerably smaller release of free endotoxin and smaller induction Jarish-Heixhaimer's reaction . The side effect stimulates inflammation and deepens SIRS by release toxins and increase of TNF-alpha.

Ned Tijdschr Geneeskd, 2001 Dec 1, 145(48), 2305 - 8
{Spondylodiscitis in 3 children; differential diagnosis and treatment}; van Henten DM et al.; Three children, a 4.5-year-old boy and two girls aged 21 months and 10 years respectively, had for several weeks to months experienced lower back pain or walking problems, two of them had an elevated sedimentation and leucocytosis . The MRI scan revealed a narrowing of the lumbal disk . Furthermore, in the case of the 10-year-old girl, Staphylococcus lugdunensis was cultured from the puncture material of the disk . After treatment she continued to experience intermittent complaints of back pain; the other children made a complete recovery . (Spondylo)discitis must be differentiated from vertebral osteomyelitis . In the case of (spondylo)discitis, immobilising the spine with a corset is the mainstay of treatment . Antibiotics are only indicated when osteomyelitis cannot be excluded . Generally, the prognosis is good.

J Mol Microbiol Biotechnol, 2002 Jan, 4(1), 1 - 10
Structure and function of pore-forming beta-barrels from bacteria; Delcour AH; Crystallographic studies of the past ten years have revealed that many outer membrane proteins and bacterial toxins are constructed on the beta-barrel motif . Two structural classes can be identified . The first class, represented by the porins, includes monomeric or multimeric proteins where each beta-barrel is formed from a single polypeptide . The second class features proteins where the beta-barrel is itself a multimeric assembly, to which each subunit contributes a few beta-strands . In addition to structural investigations, much work has also been devoted to the functional aspects of these proteins, and to the relationships between structure and function . Here we present a review of the structural and the functional properties of some of the best-studied examples of these various classes of proteins, namely the general-diffusion, specific and ligand-gated porins, multidrug efflux proteins and the staphylococcal toxin alpha-hemolysin.

Res Microbiol, 2001 Nov, 152(9), 805 - 10
A sequence variant of Staphylococcus hominis with a high prevalence of oxacillin and fluoroquinolone resistance; Fitzgibbon JE et al.; A newly identified subspecies of Staphylococcus hominis, S . hominis subsp . novobiosepticus, was found to be the cause of several invasive infections at a hospital in New Jersey . This subspecies differs from classical S . hominis, now S . hominis subsp . hominis, by the phenotypic characteristics of novobiocin resistance and the inability to ferment trehalose . DNA sequences of segments of 16S rRNA, DNA gyrase (gyrA), and DNA topoisomerase IV (grlA) genes were determined for the type strains of the 2 subspecies, and for 34 S . hominis clinical isolates . The 16S rRNA sequences of the type strains differed at 3 positions over 410 bp; the grlA sequences differed at 6 positions over 119 bp . These sequence differences define S . hominis subsp . novobiosepticus and S . hominis subsp . hominis "sequevars." Of 34 S . hominis clinical isolates, 31 were S . hominis subsp . novobiosepticus sequevars, 28 of which were resistant to both oxacillin and ciprofloxacin . The clinical microbiology laboratory, using a MicroScan system, identified 7 of the 31S . hominis subsp . novobiosepticus sequevars as S . hominis subsp . hominis on the basis of phenotypic characteristics . Three S . hominis subsp . hominis sequevars were all identified phenotypically as S . hominis subsp . hominis and were oxacillin- and ciprofloxacin-susceptible . Although the precise relationship between the S . hominis sequevars and their phenotypic subspecies remains to be determined, our results indicate that antibiotic-resistant clinical isolates of S . hominis belong almost exclusively to the S . hominis subsp . novobiosepticus sequevar.

Chir Ital, 1998 Mar-Aug, 50(2-4), 17 - 9
{Percutaneous pigtail catheter drainage of peripheral breast abscesses}; Pluchinotta AM et al.; Almost all peripheral non-lactational breast abscesses are staphylococcal in origin and so percutaneous drainage can be a valid alternative to surgical incision . Percutaneous drainage was performed in eight patients with one or more abscesses . A pigtail catheter was inserted only in foci greater than 3 cm . The results were good in 9 out of 10 purulent collections; one recurrence was observed in the smallest focus . All patients but one underwent local anesthesia and were managed as outpatients.

Microb Drug Resist, 2001 Fall, 7(3), 297 - 300
Decreased affinity of PBP3 to methicillin in a clinical isolate of Staphylococcus epidermidis with borderline resistance to methicillin and free of the mecA gene; Petinaki E et al.; Analysis of the antibiotic binding capacity of penicillin-binding proteins (PBPs) in a group of clinical isolates of Staphylococcus epidermidis suggests that the increased level of resistance to methicillin (MIC 4.0 microg/ml) in an isolate free of the mecA gene is due to the decreased binding capacity of PBP3.

J Agric Food Chem, 2002 Jan 2, 50(1), 230 - 4
Inactivation of staphylococcal enterotoxin-A with an electrolyzed anodic solution; Suzuki T et al.; Electrolyzed anodic NaCl solutions {EW+}, prepared by the electrolysis of 0.1% NaCl, have been shown to instantly inactivate most pathogens that cause food-borne disease . Elimination of food-borne pathogens does not necessarily guarantee food safety because enterotoxins produced by pathogens may remain active . We have tested whether EW+ can inactivate Staphylococcal enterotoxin A (SEA), one of the major enterotoxins responsible for food poisoning . Fixed quantities of SEA were mixed with increasing molar ratios of EW+, and SEA was evaluated by reversed-phase passive latex agglutination (RPLA) test, immunoassay, native polyacrylamide gel electrophoresis (PAGE), and amino acid analysis after 30 min incubations . Exposure of 70 ng, or 2.6 pmol, of SEA in 25 microL of PBS to a 10-fold volume of EW+, or ca . 64.6 x 10(3)-fold molar excess of HOCl in EW+, caused a loss of immuno-reactivity between SEA and a specific anti-SEA antibody . Native PAGE indicated that EW+ caused fragmentation of SEA, and amino acid analysis indicated a loss in amino acid content, in particular Met, Tyr, Ile, Asn, and Asp . Staphylococcal enterotoxin-A excreted into culture broth was also inactivated by exposure to an excess molar ratio of EW+ . Thus, EW+ may be a useful management tool to ensure food hygiene by food processing industries.

Eur J Immunol, 2002 Jan, 32(1), 50 - 8
Involvement of zinc in the binding of Mycoplasma arthritidis-derived mitogen to the proximity of the HLA-DR binding groove regardless of histidine 81 of the beta chain; Etongue-Mayer P et al.; Although our recent studies have provided the first evidence demonstrating the direct binding of Mycoplasma arthritidis-derived mitogen (MAM) to MHC class II molecules, it is not yet established how MAM interacts with these molecules . Herein, we demonstrate that MAM binds preferentially and with high affinity to HLA-DR molecules in a zinc-dependent manner . MAM's affinity (25 nM) for HLA-DR molecules is comparable to that of staphylococcal superantigens, and is slightly higher than that for murine MHC class II molecules expressed on the A20 B cell line (111 nM) . The amino acid residues located between 14 - 31 and 76 - 90 of the MAM N-terminus play a critical role in MAM / HLA-DR interactions . Histidine at position 81 of the HLA-DR beta-chain, known to be critical for binding of zinc-coordinated superantigens, is not necessary for MAM / HLA-DR interactions . The HLA-DR residues involved in MAM binding are located in the proximal binding groove of the HLA-DR molecule, where the nature of the peptide of the binding groove plays an important role in MAM / HLA-DR interaction . This is the first detailed characterization of MAM's interactions with MHC class II molecules showing a mode of interaction with HLA-DR distinct from that of other superantigens.

J Endotoxin Res, 2001, 7(6), 456 - 60
Macrophage migration inhibitory factor (MIF) modulates innate immune responses induced by endotoxin and Gram-negative bacteria; Roger T et al.; Discovered in the early 1960s as a T-cell cytokine, MIF has emerged to be an important mediator of the innate immune system . MIF was identified recently to be released by a vast array of cells, including monocytes/macrophages, T-cells, B-cells, endocrine cells and epithelial cells in response to infection and stress . Bacteria, microbial toxins and cytokines have been shown to be powerful inducers of MIF secretion by macrophages . MIF stimulates the expression of pro-inflammatory mediators by immune cells and functions to counterbalance the anti-inflammatory and immunosuppressive effects of glucocorticoids . Like TNF and IL-1, MIF plays an important role in host responses to infection . Recombinant MIF was found to exacerbate lethal endotoxemia or bacterial sepsis when co-injected with LPS or Escherichia coli in mice . Conversely, MIF knockout mice or mice treated with anti-MIF antibodies were protected from shock induced by LPS, staphylococcal exotoxins or bacterial peritonitis, even when anti-MIF therapy was started after the onset of infection . Given the central role played by MIF in innate immune responses against microbial pathogens and in the regulation of inflammatory responses, pharmacological modulation of MIF production or neutralization of MIF activity could have broad clinical applications and may offer new treatment options for the management of patients with severe sepsis or septic shock.

J Immunol, 2002 Jan 1, 168(1), 505 - 10
Decreased IL-15 may contribute to elevated IgE and acute inflammation in atopic dermatitis; Ong PY et al.; PBMC and acute skin lesions of patients with atopic dermatitis (AD) are characterized by increased IL-4 and IL-13, but decreased IFN-gamma production . This bias toward an increased Th2 cytokine profile may contribute to the elevated IgE levels and acute skin inflammation seen in AD . In this study, we examined the levels of IL-15, a Th1-like cytokine, in the PBMC and the skin lesions of AD patients . IL-15 secretion by Staphylococcal enterotoxin B-treated PBMC of AD patients was significantly lower than that of normals and psoriasis patients (p < 0.001) . Membrane-bound IL-15 expression as measured by mean fluorescence intensity and percentage of IL-15-positive cells in Staphylococcal enterotoxin B-treated monocytes of AD patients (644 +/- 49% and 12.7 +/- 0.6%, respectively) were significantly lower than that of normals (869 +/- 56% and 15.8 +/- 1.2%, respectively) and psoriasis patients (1488 +/- 217% and 22.7 +/- 0.8%, respectively; p < 0.0007 and p < 0.0001, respectively) . The membrane-bound IL-15 expression was also significantly lower in the control monocytes of AD patients compared with that in normals and psoriasis patients . There was no significant difference in the absolute number or percentage of monocytes between the study subjects . However, psoriasis skin lesions were found to have significantly more IL-15 mRNA-expressing cells (22.4 +/- 1.7) compared with that in acute AD (7.5 +/- 1.7) and chronic AD (13.7 +/- 1.7) skin lesions (p < 0.05) . IL-15 enhanced IFN-gamma production by the PBMC of AD patients (p < 0.01), but not by that of normal individuals or psoriasis patients . In addition, IL-15 was found to suppress IgE synthesis (p < 0.01) by the PBMC of AD patients . These data support the concept that reduced IL-15 expression may contribute to the pathogenesis of AD.

J Biol Chem, 2002 Feb 22, 277(8), 6247 - 53 Epub 2001 Dec 03.
Reversible and specific extracellular antagonism of receptor-histidine kinase signaling; Lyon GJ et al.; Staphylococcal pathogenesis is regulated by a two-component quorum-sensing system, agr, activated by a self-coded autoinducing peptide (AIP) . The agr system is widely divergent and is unique in that variant AIPs cross-inhibit agr activation in heterologous combinations . Cross-inhibition, but not self-activation, is widely tolerant of structural diversity in the AIPs so that these two processes must involve different mechanisms of interaction with the respective receptors . Herein, we have utilized this naturally occurring antagonism to demonstrate that both activation and inhibition are reversible and that activators and inhibitors interact at a common site on the receptor . These results suggest that molecules designed to compete with natural agonists for binding at receptor-histidine kinase sensor domains could represent a general approach to the inhibition of receptor-histidine kinase signaling.

Exp Eye Res, 2001 Nov, 73(5), 723 - 33
Superantigen presentation by human retinal pigment epithelial cells to T cells is dependent on CD2-CD58 and CD18-CD54 molecule interactions; Jorgensen A et al.; Human retinal pigment epithelial (RPE) cells are capable of presenting bacterial superantigens (SAg) to T cells in vitro by ligation of MHC class II molecules on RPE cells with the T cell receptor . The purpose of this study was to evaluate the involvement of adhesion molecules in presentation of SAg . Cultured human fetal and adult RPE cells were treated with interferon-gamma (IFN-gamma, 500 U ml(-1) for 72 hr) and afterwards pulsed with the SAg staphylococcal enterotoxin A (SEA, 500 ng ml(-1) for 2 hr) followed by coculture with freshly obtained T cells isolated from peripheral blood . Proliferation was measured by (3)H-thymidine incorporation assay . In selected experiments, either RPE or T cells were pre-treated with blocking antibodies specific for cell surface molecules . For comparison, dendritic cells were used as superantigen presenting cells for T cells . This study showed that presentation of SEA by RPE cells to resting T cells was dependent on the presence of the molecules CD2, CD58 and CD18, CD54 . The cycling status of T cells was decisive, thus resting T cells but not activated T cells were capable to proliferate in response to SEA presentation . Proliferation of T cells induced by adult RPE cells was comparable to the proliferation induced by dendritic cells at concentrations of SAg above 100 ng ml(-1), but at concentrations of SAg below 10 ng ml(-1) the response was significantly lower for SAg presented by RPE cells compared to dendritic cells . The results demonstrate that CD2-CD58 and CD18-CD54 interactions are critical for SAg presentation by RPE cells to T cells . The findings thus suggest that also presentation of peptides to resting T cells by RPE cells may be dependent upon these interactions . (C) 2001 Academic Press.

Int J Cancer, 2001 Dec 15, 94(6), 834 - 41
Superantigen enhanced protection against a weak tumor-specific melanoma antigen: implications for prophylactic vaccination against cancer; Kominsky SL et al.; B16F10 melanoma is a tumor derived from C57BL/6 mice that has been found to be poorly immunogenic and highly aggressive . Here we have shown that vaccination of mice with irradiated B16F10 cells followed by treatment with a combination of staphylococcal enterotoxins A and B (SEA/SEB) leads to significant and specific protection against subsequent challenge with viable B16F10 cells (at least 25-fold greater than a lethal dose) . Also, 75% of mice surviving over 150 days remained tumor-free after rechallenge with viable B16F10 cells, evidence of the development of strong immunologic memory . Additional studies showed increases in CD4(+) and CD8(+) T-cell populations, cytotoxic T-lymphocyte activity and interferon-gamma production, all of which may contribute to enhanced survival . Furthermore, failure to produce protection in either CD4(-/-) or CD8(-/-) T-cell knockout mice is evidence that CD4(+) and CD8(+) T cells play an essential role in induction of immunity . These results show that superantigen administration subsequent to vaccination with inactivated tumor cells results in protective antitumor immunity . Thus, prophylactic vaccination against cancer is a feasible method for arming the immune system prior to the incidence of cancer .

Eur J Immunol, 2001 Dec, 31(12), 3755 - 62
Selective induction of CCL18/PARC by staphylococcal enterotoxins in mononuclear cells and enhanced levels in septic and rheumatoid arthritis; Schutyser E et al.; Chemokines are mediators of innate and acquired immunity . CCL18, also designated pulmonary and activation-regulated chemokine (PARC), dendritic cell-derived CC chemokine-1 (DC-CK1), alternative macrophage activation-associated CC chemokine-1 (AMAC-1) and macrophage inflammatory protein-4 (MIP-4), was for the first time isolated from peripheral blood mononuclear cells (PBMC) and biochemically characterized . We found that CCL18/PARC protein is spontaneously secreted by PBMC and is selectively induced in PBMC by staphylococcal enterotoxins (SEA, SEB) and IL-4, but not by IFN-gamma and the CXCL8/IL-8 inducers lipopolysaccharide (LPS) or Concanavalin A . Human fibroblasts, chondrocytes and endothelial cells did not produce CCL18/PARC in response to inflammatory mediators such as measles virus, double-stranded RNA, LPS or IL-1beta, whereas up to 150 ng/ml of CCL2/MCP-1 was induced under these conditions . In synovial fluids from septic and rheumatoid arthritis patients, fourfold-enhanced CCL18/PARC levels (150 ng/ml) were detected compared to those in crystal-induced arthritis and osteoarthritis . In septic arthritis, the synovial levels of CCL18/PARC were fivefold higher than those of CXCL8/IL-8 . Immunochemistry revealed CD68(+) monocytes/macrophages as the main CCL18/PARC-producing cell type in both PBMC and arthritic synovial tissue . In addition, CD1a(+) blood dendritic cells expressed CCL18/PARC . These findings suggest that monocytic cells respond to Gram-positive bacterial infection by the production of CCL18/PARC in the synovial cavity.

J Acquir Immune Defic Syndr, 2001 Dec 15, 28(5), 429 - 36
Evaluation of immune activation in HIV-infected and uninfected African individuals by single-cell analysis of cytokine production; Lukwiya M et al.; Immune activation has been observed in HIV-infected and uninfected Africans, among whom it is thought to modify interaction between the immune system and HIV . To characterize this phenomenon accurately, in-depth immunologic analyses were performed in a rural African population . Freshly drawn peripheral blood mononuclear cells (PBMCs) of HIV-infected African (from Gulu, Uganda) and Italian antiviral-naive patients and those of uninfected Ugandan and Italian study subjects were analyzed . Individuals were matched for age and sex and determined to be free from parasitic infections . Intracellular cytokines were measured in mitogen (M)- and gp160 peptides + staphylococcal enterotoxin B and alpha CD28 (env)-stimulated T lymphocytes . Interferon (IFN)-gamma-producing CD8(+) T cells were quantified in an enzyme-linked immunosorbent assay . Results showed that M-stimulated production of interleukin (IL)-10 and tumor necrosis factor (TNF)-alpha increases in CD4(+) and CD8(+) cells of African infected patients and uninfected study subject; and that env-stimulated IL-10 and TNF-alpha production is increased in CD8(+) T lymphocytes of African HIV-infected patients . M- and env-stimulated IFN-gamma-producing CD8(+) T cells were reduced in African participants and not increased by preincubation with alpha IL-10 monoclonal antibody . This is the first set of data that has reported immune activation in rural Africa by single-cell analysis of cytokine production . These results help in defining the immunologic background to be considered in the design of therapeutic and vaccine-based approaches to HIV infection in an African setting.

J Vasc Surg, 2001 Dec, 34(6), 983 - 5
Femoral artery infections associated with percutaneous arterial closure devices; Johanning JM et al.; Hemostasis obtained by manual compression after femoral artery catheterization results in consistently low rates of major complications . A rare complication of femoral artery catheterization is arterial infection . Its occurrence after diagnostic angiography using manual compression has not been reported . We report two cases of femoral arterial infection after uneventful diagnostic catheterization in nonimmunocompromised patients using the Perclose percutaneous arterial closure device . Our cases are representative of Perclose associated infections, with delayed presentation of a staphylococcal arterial infection requiring arterial debridement and reconstruction . This article indicates that Perclose use carries a risk of severe arterial infection . Surgeons should be aware of the potential infectious complications associated with Perclose use and the need for aggressive treatment.

Protein Sci, 2002 Jan, 11(1), 82 - 91
Early formation of a beta hairpin during folding of staphylococcal nuclease H124L as detected by pulsed hydrogen exchange; Walkenhorst WF et al.; Pulsed hydrogen exchange methods were used to follow the formation of structure during the refolding of acid-denatured staphylococcal nuclease containing a stabilizing Leu substitution at position 124 (H124L SNase) . The protection of more than 60 backbone amide protons in uniformly (15)N-labeled H124L SNase was monitored as a function of refolding time by heteronuclear two-dimensional NMR spectroscopy . As found in previous studies of staphylococcal nuclease, partial protection was observed for a subset of amide protons even at the earliest folding time point (10 msec) . Protection indicative of marginally stable hydrogen-bonded structure in an early folding intermediate was observed at over 30 amide positions located primarily in the beta-barrel and to a lesser degree in the alpha-helical domain of H124L SNase . To further characterize the folding intermediate, protection factors for individual amide sites were measured by varying the pH of the labeling pulse at a fixed refolding time of 16 msec . Protection factors >5.0 were observed only for amide positions in a beta-hairpin formed by strands 2 and 3 of the beta-barrel domain and a single site near the C-terminus . The results indicate that formation of stable hydrogen-bonded structure in a core region of the beta-sheet is among the earliest structural events in the folding of SNase and may serve as a nucleation site for further structure formation.

Eur J Biochem, 2001 Dec, 268(24), 6390 - 401
Pyridinyl polythiazole class peptide antibiotic micrococcin P1, secreted by foodborne Staphylococcus equorum WS2733, is biosynthesized nonribosomally; Carnio MC et al.; Recently, foodborne Staphylococcus equorum WS2733 was isolated from a French red smear cheese on account of its strong inhibitory activity against Gram-positive pathogens such as Listeria . The antagonistic substance was identified as macrocyclic peptide antibiotic micrococcin P1, which had previously not been reported for the genus Staphylococcus . Micrococcin P1, also a potent inhibitor of the malaria parasite Plasmodium falciparum, is structurally related to thiostrepton, thiocillins and nosiheptide . Although all of these peptide antibiotics have been known for quite a long time, their mode of biosynthesis had not been determined in detail yet . By using degenerated PCR, a gene fragment encoding a nonribosomal peptide synthetase (NRPS) could be amplified from S . equorum . The corresponding chromosomal locus was disrupted by insertional mutagenesis, and it could be shown that all mutants obtained displayed a micrococcin P1-deficient phenotype . Sequence analysis of a coherent 2.8-kb fragment revealed extensive homology to known NRPSs, and allowed the assignment of the domain organization 'condensation-adenylation-thiolation-condensation'; an arrangement predicted only for two loci within the presumably 14-modular, 1.6-MDa biosynthetic NRPS template . Biochemical characterization of the adenylation domain exhibited selectivity for the substrate amino-acid threonine . All of these data substantiate that the macrocyclic peptide antibiotic is biosynthesized nonribosomally, and provide the basis for the characterization of the entire biosynthetic gene cluster . The biosynthetic machinery of micrococcin will serve as a model system for structurally related, pharmacologically important pyridinyl polythiazole class peptide antibiotics . Furthermore, this knowledge will enable the manipulation of its NRPS template, which in turn may grant the targeted engineering of even more potent anti-listerial and anti-malaria drugs.

Biochemistry, 2001 Dec 18, 40(50), 15280 - 9
Packing is a key selection factor in the evolution of protein hydrophobic cores; Chen J et al.; The energy derived from optimized van der Waals interactions in closely packed, folded proteins has been proposed to be of similar energetic magnitude to hydrophobicity in stabilizing the native state . If packing is this energetically important, it should influence the evolution of protein core sequences . To test this hypothesis, the occurrence of various amino acid side chains in the major hydrophobic core of staphylococcal nuclease and 42 homologous proteins was determined . Most such positions in this protein family are usually isoleucine, leucine, or valine . Previously we have constructed and measured the stabilities of 12 single, 44 double, 64 triple, and 32 quadruple mutants, representing all possible permutations of these three side chains at two overlapping sets of four positions in the core of staphylococcal nuclease . The stabilities and interaction energies of those mutants with various combinations of the most common, or consensus, sequence were compared to the stabilities of all other mutants . Mutants which had the consensus side-chain combinations were not necessarily the most stable, but usually were found to have the best interaction energies . In other words, these proteins were far more stable than would be predicted from simply summing the observed energetic effects of the component single mutations, apparently reflecting particularly favorable packing interactions that are possible for the most common side chains . An additional 12 mutants which tested possible alternate explanations of the results were constructed . The stabilities and interaction energies of these mutants also support the conclusion that packing is a crucial determinant guiding the sequence evolution of protein cores.

Eur J Pharmacol, 2001 Nov 30, 432(1), 107 - 19
The anti-inflammatory activities of cannabinoid receptor ligands in mouse peritonitis models; Smith SR et al.; In this report, we describe experiments in which cannabinoid receptor ligands were evaluated for effects on the development of a peritoneal inflammation when elicited in mice with thioglycollate broth or staphylococcus enterotoxin A . The cannabinoid receptor agonists {(-)-11-hydoxy-Delta(8) tetrahydrocannabinol-dimethylheptyl} (HU-210) and {(R)-(+)-{2,3-dihydro-5-methyl-3-{(4-morpholinyl)methyl{pyrrolo{1,2,3-de}1,4-benzoxazin-6-yl}(1-naphthalenyl) methanone} (WIN 55212-2) blocked the migration of neutrophils into the peritoneal cavity in response to these inflammatory stimuli . This effect was caused by a delay in the production of the neutrophil chemoattractants, KC and macrophage inflammatory protein-2 . HU-210 and WIN 55212-2 blocked neutrophil chemokines and neutrophil migration whether administered subcutaneously (s.c.) or intracerebroventricularly (i.c.v.) . Their modulatory effects on the inflammation were antagonized by centrally administered {N-(piperdin-1-yl)-5-(4-chloropheny)-1-(2,4-dichloropheny)-4-methyl-1H-pyrazole-3-carboxamide hydrochloride} (SR141716A), a selective cannabinoid CB(1) receptor antagonist . This latter observation, and the ability of the cannabinoid receptor agonists to suppress the peritoneal inflammation at relatively low doses when administered i.c.v., indicated a role for central cannabinoid CB(1) receptors in the anti-inflammatory activities of HU-210 and WIN 55212-2 . The cannabinoid receptor agonists had no effect on monocyte migration elicited by thioglycollate, despite their ability to suppress monocyte chemotactic protein-1 levels in lavage fluids . The cannabinoid CB(2) receptor antagonist, {N-{(1S)-endo-1,3,3-trimethylbicyclo{2.2.1}heptan-2-yl}5-(4-choro-3 methylphenyl)-1-(4-methylbenzyl)pyrazole-3-carboxamide} (SR144528) inhibited the peritoneal inflammation in a manner analogous to that of HU-210 and WIN 55212-2 when administered i.c.v., but it did not appear to act through central cannabinoid CB(1) receptors . The present results add to the body of literature indicating that cannabinoid receptor ligands have diverse anti-inflammatory properties.

Clin Imaging, 2001 Nov-Dec, 25(6), 396 - 9
Internal mammary artery pseudoaneurysms complicating chest wall infection in children: diagnosis and endovascular therapy; Deshmukh H et al.; Mycotic internal mammary artery (IMA) pseudoaneurysms are sparsely reported in medical literature . We report imaging findings of IMA pseudoaneurysms secondary to chest wall abscesses (staphylococcal and tuberculous) in two children . Both children were successfully treated by endovascular method thus obviating the need for surgery.

J Neuroimmunol, 2001 Dec 3, 121(1-2), 88 - 101
Single-cell analysis of cytokine production shows different immune profiles in multiple sclerosis patients with active or quiescent disease; Clerici M et al.; Peripheral blood mononuclear cells of multiple sclerosis (MS) patients were stimulated with myelin basic protein (MBP) together with anti-CD28 monoclonal antibody and staphylococcal enterotoxin B to optimize cytokine production by antigen-specific cells . Type 1 (IL-2, IL-12, IFNgamma) and pro-inflammatory (TNFalpha, IL-1beta, IL-6) cytokines were augmented in CD4+, CD8+, and CD14+ cells of acute MS patients and of patients undergoing disease reactivation . These cytokines were reduced in IFNbeta-treated and in stable MS patients; type 2 cytokines (IL-4, IL-10) were increased in these patients . Similar immune profiles are seen in MS patients in whom remission is naturally or pharmacologically (IFNbeta) achieved . Cytokine alterations are particularly evident in CD14+ cells, underlying their critical role in the modulation of the immune response.

Transfusion, 2001 Nov, 41(11), 1351 - 5
First results using automated epifluorescence microscopy to detect Escherichia coli and Staphylococcus epidermidis in WBC-reduced platelet concentrates; Seaver M et al.; BACKGROUND: Many methods have been tested for the detection of bacterial contamination in platelets . However, only those using molecular biology or cell culturing consistently detect contamination at levels below 10(5) bacteria per mL . This report describes the initial investigation into an alternative method that offers the possibilities of high sensitivity and rapid response while using available laboratory equipment and supplies . This method relies on a fluorescent nucleic acid stain, which preferentially stains bacteria but not platelets, and automated epifluorescence microscopy for rapid analysis . Measurements in WBC-reduced platelet concentrates (PCs) contaminated with bacteria are reported at concentrations between 10(3) and 10(6) bacteria per mL . STUDY DESIGN AND METHODS: Staphylococcus epidermidis or Escherichia coli was inoculated into aliquots of WBC-reduced PCs on Days 2 through 5 of storage . Bacterially inoculated and control PCs were stained, platelets and residual WBCs were lysed, and 200 microL of sample was filtered onto black polycarbonate filters . All preparations were done in triplicate . An automated epifluorescence microscope examined approximately 2 percent of the area of each filter and used image analysis to select the fluorescent particles that should be counted as bacteria . RESULTS: Samples containing 3 to 5 x 10(3) bacteria per mL produced about three times as many fluorescent particles classified as bacteria as the controls . Lower concentrations of S . epidermidis were detected because of higher fluorescence intensity . Simultaneous preparation of six samples requires about 35 minutes . Analysis of each prepared sample takes 10 minutes, for a total preparation and analysis time of about 95 minutes for 6 samples . CONCLUSION: Low concentrations (<5 x 10(3) bacteria/mL) of deliberately inoculated S . epidermidis or E . coli can be measured quickly in WBC-depleted PCs by using a fluorescent nucleic acid stain, differential lysis, and automated microscopy . Continued refinement of the method, studies employing other bacterial strains, and further validations of assay performance are warranted.

Rheum Dis Clin North Am, 2001 Nov, 27(4), 833 - 48
Are antineutrophil cytoplasmic antibody-associated vasculitides pauci-immune?
Brons RH, Kallenberg CG, Tervaert JW.
The role of ICs in ANCA-associated vasculitides remains controversial . The ANCA-associated vasculitides are described as being pauci-immune . We hypothesize that the absence of ICs is a result of an exaggerated inflammatory response caused by the presence of ANCAs . We present evidence indicating that ICs may play a role in the initiation or relapses of the disease . The nature of the involved antigen(s) is not yet known . Possible candidates are reviewed and include ANCA antigens, AECA antigens, and staphylococcal antigens.

Biophys J, 2001 Dec, 81(6), 3489 - 502
pH corrections and protein ionization in water/guanidinium chloride; Garcia-Mira MM et al.; More than 30 years ago, Nozaki and Tanford reported that the pK values for several amino acids and simple substances in 6 M guanidinium chloride differed little from the corresponding values in low salt (Nozaki, Y., and C . Tanford . 1967 . J . Am . Chem . Soc . 89:736-742) . This puzzling and counter-intuitive result hinders attempts to understand and predict the proton uptake/release behavior of proteins in guanidinium chloride solutions, behavior which may determine whether the DeltaG(N-D) values obtained from guanidinium chloride-induced denaturation data can actually be interpreted as the Gibbs energy difference between the native and denatured states (Bolen, D . W., and M . Yang . 2000 . Biochemistry . 39:15208-15216) . We show in this work that the Nozaki-Tanford result can be traced back to the fact that glass-electrode pH meter readings in water/guanidinium chloride do not equal true pH values . We determine the correction factors required to convert pH meter readings in water/guanidinium chloride into true pH values and show that, when these corrections are applied, the effect of guanidinium chloride on the pK values of simple substances is found to be significant and similar to that of NaCl . The results reported here allow us to propose plausible guanidinium chloride concentration dependencies for the pK values of carboxylic acids in proteins and, on their basis, to reproduce qualitatively the proton uptake/release behavior for the native and denatured states of several proteins (ribonuclease A, alpha-chymotrypsin, staphylococcal nuclease) in guanidinium chloride solutions . Finally, the implications of the pH correction for the experimental characterization of protein folding energetics are briefly discussed.

FEMS Immunol Med Microbiol, 2001 Oct, 31(3), 197 - 202
Binding of human clusterin by Staphylococcus epidermidis; Li DQ et al.; To hypothesise that Staphylococcus epidermidis may possess clusterin receptor(s), bacterial binding of human clusterin was determined . In a fluid phase, the binding was markedly influenced by culture medium and three out of 12 S . epidermidis strains grown on ISO-sensitest agar expressed clusterin-binding ability . S . epidermidis J9P, selected for further study, showed saturable binding of iodine-labelled clusterin, and the binding was only inhibited by unlabelled clusterin . The binding was sensitive to protease treatment . Scatchard plot indicated one single class of binding sites (K(d)=104.2 nM) . None of the S . epidermidis strains bound immobilised clusterin . These data imply that ligand-receptor interaction exists between S . epidermidis and clusterin in fluid phase, but the domain(s) recognised by bacteria may have been occluded when clusterin was adsorbed on a surface.

Cell Immunol, 2001 Aug 25, 212(1), 63 - 70
Characterization of a novel H2A(-)E+ transgenic model susceptible to heterologous but not self thyroglobulin in autoimmune thyroiditis: thyroiditis transfer with Vbeta8+ T cells; Yan Y et al.; Recently we reported on a novel H2E transgenic, IA-negative model of experimental autoimmune thyroiditis (EAT) that excludes reactivity to self in its susceptibility pattern to heterologous thyroglobulin (Tg) . In conventional, susceptible mouse strains, EAT is inducible with both homologous and heterologous Tg; e.g., human (h)Tg shares conserved thyroiditogenic epitopes with mouse (m)Tg . However, when an H2Ea(k) transgene is introduced into class II-negative B10.Ab(0) mice, which express neither surface IA (mutant Abeta-chain) nor surface IE (nonfunctional Ea gene), the resultant H2E(b) molecules are permissive for EAT induction by hTg, but not self mTg . Also, the hTg-primed cells do not cross-react with mTg . To explore this unique capacity of E+B10.Ab(0) mice to distinguish self from nonself Tg, we have developed T cell lines to examine the T cell receptor repertoire and observed a consistent Vbeta8+ component after repeated hTg stimulation . Enrichment and activation of Vbeta8+ T cells by either superantigen staphylococcal entertoxin B or anti-Vbeta8 in vitro enabled thyroiditis transfer to untreated A-E+ recipients, similar to hTg activation . Vbeta8+ T cells isolated by FACS from hTg-immunized mice also proliferated to hTg in vitro . These studies support the contribution of Vbeta8 genes to the pathogenicity of hTg in this H2A-E+ transgenic model.

Cell Immunol, 2001 Aug 25, 212(1), 35 - 43
Roles of I-E molecule and CD28 costimulation in induction of suppression by staphylococcal enterotoxin B in vivo; Hsu LJ et al.; Exposure to bacterial superantigens leads to the induction of a subsequent state of immune hyporesponsiveness . Using a transwell coculture system, a previous report demonstrated that splenocytes from staphylococcal enterotoxin B (SEB)-injected BALB/c mice secreted soluble mediators to suppress the proliferative response of naive syngeneic splenocytes to SEB stimulation . We show in the present study that, in contrast to the suppressive effect induced by SEB in BALB/c (H-2(d) haplotype), MRL(+/+), and MRL-lpr/lpr (H-2(k)) mice, SEB-primed splenocytes from I-E(-) strains such as B6, B10, A.BY (H-2(b)), and A.SW (H-2(s)) mice failed to inhibit the CD25 expression and the proliferative activity of their syngeneic naive responder splenocytes . Further results revealed that the SEB-primed cells from BALB/c, but not B6, mice inhibited the CD25 expression and proliferation of naive responder cells from either BALB/c or B6 mice, indicating the critical regulatory role of the effector cells . Unlike SEB, staphylococcal enterotoxin A induced profound suppression in both BALB/c and B6 mice . Moreover, the suppressive competence of SEB-primed splenocytes was diminished in CD28-deficient BALB/c mice . Taken together, our results indicate that when SEB is used as a stimulator in vivo, both the I-E molecule and CD28 costimulation are required for the induction of regulatory cells bearing suppressive activity . (c) 2001 Elsevier Science.

J Biol Inorg Chem, 2001 Oct, 6(8), 757 - 62
Cooperative zinc binding in a staphylococcal enterotoxin A mutant mimics the SEA-MHC class II interaction; Hakansson M et al.; The structure of a mutant form of staphylococcal enterotoxin A (SEA) has been determined to 2.1 A resolution . The studied SEA substitution H187-->A187 (SEAH187A) leads to an almost 10-fold reduction of the binding to major histocompatibility complex (MHC) class II . H187 is important for this interaction since it coordinates Zn2+ . The zinc ion is thought to hold MHC class II and SEA together in a complex . Interestingly, only one of two molecules in the asymmetric unit binds Zn2+ . H225, D227, a water molecule, and H44 from a symmetry-related molecule ligate Zn2+ . The symmetry-related histidine is necessary for this substituted Zn2+ site to bind to Zn2+ at low zinc concentration (no Zn2+ added) . Since a water molecule replaces the missing H187, H44 binds Zn2+ at the position where betaH81 from MHC class II probably will bind . Dynamic light scattering analysis reveals that in solution as well as in the crystal lattice the SEA(H187A) mutant forms aggregates . The substitution per se does not cause aggregation since wild-type SEA also forms aggregates . Addition of EDTA reduces the size of the aggregates, indicating a cross-linking function of Zn2+ . In agreement with the biological function, the aggregation is weak (i.e . not revealed by gel filtration) and non-specific.

J Interferon Cytokine Res, 2001 Oct, 21(10), 793 - 6
Upregulation of IFN-gamma and soluble interleukin-2 receptor release and altered serum cortisol and prolactin concentration during general anesthesia; Brand JM et al.; The effects of surgery, surgical stress, and anesthesia compromise the optimal function of the immune system . Recent studies demonstrate the influence of anesthesia on the immune response by modulation of neural-immune interactions . To evaluate the immunologic effects of general anesthesia with the hypnotic agent propofol and the opioid fentanyl, two drugs used frequently in anesthesia, we studied 30 patients undergoing elective orthopedic surgery before and during narcosis . We found a significant enhancement of interferon-gamma (IFN-gamma) and soluble interleukin-2 receptor (sIL-2R) release in lipopolysaccharide (LPS)-stimulated whole blood cultures after induction of anesthesia . Similar results were observed in cultures stimulated with polyclonal T cell activators, such as staphylococcal enterotoxin B (SEB) and phytohemagglutinin (PHA) . IL-1beta and IL-8 release was not affected, but the anti-inflammatory cytokine IL-10 decreased after skin incision . Serum prolactin significantly increased immediately after induction of anesthesia, whereas serum cortisol levels declined . Our results point to enhanced proinflammatory T lymphocyte and natural killer (NK) cell activity, probably caused by prolactin and cortisol modulation in the serum . This may disturb the balance of human proinflammatory and anti-inflammatory pathways during surgery and general anesthesia.

Microb Pathog, 2001 Nov, 31(5), 205 - 12
Interaction of coagulase-negative Staphylococcus species with bovine mammary epithelial cells; Almeida RA et al.; Three coagulase-negative Staphylococcus species (CNS) (Staphylococcus epidermidis, Staphylococcus xylosus and Staphylococcus hyicus), from the milk of cows with mastitis, were used to evaluate adherence to and internalization by bovine mammary epithelial cells, and to investigate involvement of host cell signal transduction and host cell cytoskeleton rearrangement on internalization of CNS . S . xylosus showed highest adherence and internalization values of the species evaluated . Host cell cytoskeleton polymerization and protein kinase (PK) phosphorylation were required for internalization of CNS . Both protein kinase C (PKC) and tyrosine kinase (TPK) pathways were involved, but internalization of S . xylosus occurred preferentially through epidermal growth factor TPK activity . S . epidermidis and S . hyicus seemed to exploit other TPK pathways . Results of this study showed that S . xylosus, S . hyicus and S . epidermidis were able to adhere and internalize bovine mammary cells in a process that appeared to be receptor(s) mediated and exploited host signal transduction and cytoskeleton to induce an uptake signal .

Infect Immun, 2001 Dec, 69(12), 7544 - 9
Temporal sequence and kinetics of proinflammatory and anti-inflammatory cytokine secretion induced by toxic shock syndrome toxin 1 in human peripheral blood mononuclear cells; Kum WW et al.; The staphylococcal superantigen toxic shock syndrome toxin 1 (TSST-1) induces massive cytokine production, which is believed to be the key factor in the pathogenesis of TSS . The temporal sequence and kinetics of both proinflammatory and anti-inflammatory cytokines induced by TSST-1 in human peripheral blood mononuclear cells were investigated . A panel of loss-of-function single-amino-acid-substitution mutants of TSST-1, previously demonstrated to be defective in either major histocompatibility complex (MHC) class II binding (G31R) or T-cell receptor (TCR) interaction (H135A, S14N), was studied in parallel to further elucidate the mechanisms of cytokine secretion . Wild-type recombinant (WT r) TSST-1 induced a biphasic pattern of cytokine secretion: an early phase with rapid release of proinflammatory cytokines (especially gamma interferon, interleukin-2 {IL-2}, and tumor necrosis factor alpha {TNF-alpha}) within 3 to 4 h poststimulation, and a later phase with more gradual production of both proinflammatory (IL-1beta, IL-12, and TNF-beta) and anti-inflammatory (IL-6, IL-10) cytokines within 16 to 72 h poststimulation . G31R, which is defective in MHC class II binding, induced a cytokine profile similar to that of WT rTSST-1, except that secretion of the early-phase proinflammatory cytokines was delayed and production of IL-1beta and IL-12 was markedly reduced . In contrast, mutant toxins defective in TCR interaction either demonstrated complete absence of any cytokine secretion during the entire observation period (H135A) or resulted in complete abolishment of IL-2 and other early-phase proinflammatory cytokines, while secretion of IL-10 appeared unaffected (S14N) . Neither WT rTSST-1 nor the mutant toxins induced IL-4 or transforming growth factor beta . Our data indicate that effective TCR interaction is critical for the induction of the early-phase proinflammatory cytokine response, thus underscoring the importance of T-cell signaling in TSS.

Biochemistry, 2001 Nov 20, 40(46), 14012 - 9
Higher-order packing interactions in triple and quadruple mutants of staphylococcal nuclease; Chen J et al.; Sixty-four triple and 32 quadruple mutants were constructed in the core of staphylococcal nuclease . This is the first time that a large number of multiple mutants with all possible variations and all possible lower-order mutants has been systematically constructed in any protein core . Stabilities were determined by solvent denaturation . The energetic effects of these multiple mutants have been analyzed in combination with the stability data from the component single and double mutants . It was found that most of the stability changes in triple and quadruple mutants cannot be correctly predicted from stability effects of component single mutants . However, if the interaction energy between pairs of side chains in the component double mutants is taken into account, correct stability prediction can be made for most triple and quadruple mutants . The data further show that while packing interactions unique to triple and quadruple mutations do occur, they are of much less energetic significance than interactions between pairs of residues . The results presented here show that the packing of a protein interior can be closely approximated in most cases as a series of short-range, nearest-neighbor interactions . This has profound implications for rational protein design and structure prediction.

Biochemistry, 2001 Nov 20, 40(46), 14004 - 11
Energetics of side chain packing in staphylococcal nuclease assessed by systematic double mutant cycles; Chen J et al.; All 44 possible double mutant permutations of isoleucine, leucine, and valine were constructed in 11 pairings of six sites in the core of staphylococcal nuclease . The stabilities of these mutants were determined by guanidine hydrochloride denaturation . Comparison of the stabilities of all double mutants with those expected from addition of the corresponding single mutants showed that the effects of the two single mutations are energetically independent of each other in 30 of the double mutants . However, a substantial minority, 14, of the double mutants have stability effects that are not additive . In these cases, it appears that direct van der Waals contacts between the two side chains are present . The requirement of direct van der Waals contact for the interdependence of mutational stability effects is somewhat surprising in light of results previously reported by others . In addition, it was found that double mutants that did not alter or lower the overall number of atoms in the core and that showed nonadditive behavior were more stable than expected from addition of the effects of the corresponding single mutants . A net increase in the number of atoms in the core usually, but not always, resulted in a mutant that was less stable than expected . In contrast to previous staphylococcal nuclease double mutants, energetically significant changes to the denatured state do not appear to be occurring in these packing mutants . These conclusions imply that attempts to engineer protein stability based on single mutant data will be generally successful if overall core size is preserved and if residues are not in van der Waals contact.

Biochemistry, 2001 Nov 20, 40(46), 13998 - 4003
Energetics of side chain packing in staphylococcal nuclease assessed by exchange of valines, isoleucines, and leucines; Holder JB et al.; To examine the importance of side chain packing to protein stability, each of the 11 leucines in staphylococcal nuclease was substituted with isoleucine and valine . The nine valines were substituted with leucine and isoleucine, while the five isoleucines, previously substituted with valine, were substituted with leucine and methionine . These substitutions conserve the hydrophobic character of these side chains but alter side chain geometry and, in some cases, size . In addition, eight threonine residues, previously substituted with valine, were substituted with isoleucine to test the importance of packing at sites normally not occupied by a hydrophobic residue . The stabilities of these 58 mutant proteins were measured by guanidine hydrochloride denaturation . To the best of our knowledge, this is the largest library of single packing mutants yet characterized . As expected, repacking stability effects are tied to the degree of side chain burial . The average energetic cost of moving a single buried methyl group was 0.9 kcal/mol, albeit with a standard deviation of 0.8 kcal/mol . This average is actually slightly greater than the value of 0.7-0.8 kcal/mol estimated for the hydrophobic transfer energy of a methylene from octanol to water . These results appear to indicate that van der Waals interactions gained from optimal packing are at least as important in stabilizing the native state of proteins as hydrophobic transfer effects.

Eur J Dermatol, 2001 Nov-Dec, 11(6), 549 - 53
Erythromycin-resistance of cutaneous bacterial flora in acne; Dreno B et al.; Some studies have raised the problem of an increase of bacterial resistance in acne patients . This study was carried out in France, where no previous studies about resistance to erythromycin has been performed, on 40 patients with mild to moderately severe acne . Microbiological samples were obtained by using Williamson and Kligman method . This study showed that the prevalence of bacterial resistance to erythromycin was 95% for Staphylococcus epidermidis strains and 52% for Propionibacterium acnes strains . Resistant strains were more frequent in patients with predominantly inflammatory lesions (papules and pustules) . The influence of previous or current treatment with erythromycin was also studied and showed that even patients without any previous use of erythromycin had resistant strains for Propionibacterium acnes (42%) . In addition the minimum inhibitory concentrations (MIC) for zinc were evaluated . All the Propionibacterium acnes strains tested were inhibited at concentrations less or equal to 512 mug/ml of zinc . However, zinc combined with erythromycin in vitro did not modify the erythromycin MIC . This first French study on bacterial resistance to erythromycin in acne patients confirms the results of studies performed in other countries.

Spine, 2001 Oct 1, 26(19), E463 - 7
Halo pin intracranial penetration and epidural abscess in a patient with a previous cranioplasty: case report and review of the literature; Papagelopoulos PJ et al.; STUDY DESIGN: Report of a patient with an epidural abscess after halo pin intracranial penetration at the site of a previous cranioplasty . OBJECTIVES: To report a rare case of intracranial penetration at the site of a previous cranioplasty associated with epidural abscess, and to discuss the diagnostic and therapeutic approach to its management . SUMMARY OF BACKGROUND DATA: The most serious complications associated with use of halo device occur when pins penetrate the inner table of the skull, resulting in cerebrospinal fluid leak and rarely in an intracranial abscess . However, no mention of intracranial halo pin penetration at the site of a previous cranioplasty was found in the literature . METHODS: A 64-year-old man with ankylosing spondylitis had a halo vest placed for management of a fracture dislocation through the C5-C6 intervertebral disc space associated with left C6 radiculopathy . One week later, the patient experienced fever and headache associated with pain, redness, and drainage at the site of the insertion of the left posterior pin . Computed tomography of the brain showed a 1.5-cm intracranial penetration of the halo pin through a previous cranioplasty of the temporal bone, associated with epidural abscess and cerebral edema in the left temporoparietal lobe . The pins and the halo vest were removed, the pin site was cleaned, and a Philadelphia cervical collar was applied . Staphylococcus epidermidis grew on the culture of drainage from the pin site . The patient started immediate intravenous antibiotic treatment for 2 weeks, followed by oral antibiotics for 2 additional weeks . RESULTS: The patient had gradual improvement of his symptoms within the first 48 hours . At the latest follow-up visit, he had fully recovered and his fracture had healed . CONCLUSIONS: The halo device should not be used for patients with a previous cranioplasty, especially if the pins cannot be inserted at other safe areas of the skull . A thorough medical history and physical examination of the skull are important before the application of a halo device . Computed tomography of the skull may be necessary before elective halo application for patients with concomitant head trauma, confusion, or intoxication and for patients with a previous cranioplasty to ascertain the safest pin sites.

Pediatr Neurosurg, 2001 Oct, 35(4), 205 - 10
The treatment of cerebrospinal fluid shunt infections . Results from a practice survey of the American Society of Pediatric Neurosurgeons; Whitehead WE et al.; It is our impression that the management strategy for infected cerebrospinal fluid (CSF) shunts varies significantly among pediatric neurosurgeons . The purpose of this paper is to present the results of a practice survey on the treatment of shunt infections which was distributed to all active members of the American Society of Pediatric Neurosurgeons (ASPN) . Eighty-four of 129 ASPN members (65%) responded to the survey . Most ASPN members remove the shunt and place an external ventricular drain (EVD) to treat Staphylococcus epidermidis (59.5%), S . aureus (64.3%) and gram-negative rod infections (67.9%) . The second most common method of treatment was externalization of the shunt (33.3, 29.8 and 25%, respectively) . The duration of antibiotic treatment was extremely variable . When the shunt was removed and an EVD inserted, the duration of antibiotic treatment for S . epidermidis and S . aureus ranged from 5 to 21 total days (2-21 days of sterile cultures) . For gram-negative rod infections treated with shunt removal and an EVD, the total duration of antibiotic therapy ranged from 5 to 24 days (2-37 days of sterile cultures) . The majority of ASPN members remove the infected CSF shunt and place an EVD for the management of shunt infections . Significant variation exists in the duration of antibiotic therapy . Determining the most effective duration of antibiotic therapy in an effort to shorten hospitalization and minimize complications without sacrificing efficacy will require further study .

J Immunol Methods, 2001 Nov 1, 257(1-2), 83 - 92
Development of a time-resolved immunofluorometric assay for quantitation of mucosal and systemic antibody responses; Hale ML et al.; We developed a solid phase immunoassay that measured mucosal and systemic antibody responses from mice inoculated with either a staphylococcal enterotoxin B vaccine (SEBv) or noninfectious virus-like particles (VLP) of lentiviral origin . The assay used time-resolved fluorescence (TRF) with affinity-purified goat anti-mouse IgA and IgG conjugated to samarium and europium chelates, respectively . By employing these fluorogenic conjugates with different spectral emissions, IgA and IgG specific for SEB or VLP were readily detected in serum and saliva from mice inoculated intranasally . The TRF assay detected antigen-specific IgA in saliva 10 min after the addition of enhancement solution, while a conventional alkaline phosphatase-based assay for salivary IgA required 18 h after substrate addition . The TRF assay also provided a significantly higher signal-to-noise ratio and exhibited greater sensitivity . TRF assays detected both IgA and IgG in the same well, thereby reducing sample and reagent requirements.

Langenbecks Arch Surg, 2001 Aug, 386(5), 328 - 32
In vitro adherence and accumulation of Staphylococcus epidermidis RP 62 A and Staphylococcus epidermidis M7 on four different bone cements; Konig DP et al.; Bacterial resistance of Staphylococcus epidermidis, a serious pathogen of implant-related infections, to antibiotics is related to the production of a glycocalyx slime that impairs antibiotic access and the killing by host defense mechanisms . In vitro studies of different bone cements containing antibiotics, developed for the prevention of biomaterial-associated infection, could not always demonstrate complete eradication of biomaterial-adherent bacteria . We have investigated four different bone cements in regard to bacterial accumulation of a slime-producing strain RP 62 A and its isogenic mutant M7 lacking the ability to produce exopolysaccharide slime using a bacterial adhesion assay and modified Kirby-Bauer technique . A significant effect of exopolysaccharide production for the accumulation on bone cement could be demonstrated . The gentamicin/clindamycin bone cement was the only tested biomaterial that produced a large zone of bacterial inhibition in the inoculated area adjacent to the biomaterial . The bacterial adhesion was not reduced significantly and there was no correlation between zones of inhibition on blood agar plates and the quantitative adhesion assay . The clinical efficacy of the gentamicin/clindamycin bone cement must be proven in vivo.

Neuropharmacology, 2001 Dec, 41(7), 811 - 8
Involvement of brain glutamate release in pyrogenic fever; Huang WT et al.; Whether the glutamate release in the organum vasculosum laminae terminalis (OVLT) is attributable to genesis of a pyrogenic fever is unclear . The lack of information led us to evaluate the changes in glutamate concentrations of OVLT during the fever induced by staphylococcal enterotoxin A (SEA) in unanesthetized rabbits . Both the OVLT concentrations of glutamate and the colonic temperatures were simultaneously monitored during systemic injection of SEA, MK801 (an N-methyl-D-aspartate (NMDA) receptor channel blocker), ketamine (an NMDA receptor channel blocker), or normal saline . The extracellular dialysates in the brain were collected using a microdialysis probe previously placed in the OVLT region . The concentrations of glutamate in the microdialysates were measured by a high-pressure liquid chromatography in combination with a fluorescence detector . Systemic administration of SEA (30 ng x kg(-1) I.V.) increased both the concentrations of glutamate in the OVLT and the colonic temperatures . Glutamate appeared to rise slightly earlier than body temperature . Pretreatment or posttreatment with MK801 or ketamine significantly attenuated the SEA-induced augmenting glutamate release in the OVLT and fever in rabbits . The suppression of glutamate release appeared to start slightly earlier than temperature decline . In addition, the SEA-induced fever could be mimicked by direct injection of glutamate or SEA into the OVLT area . The fever induced by intra-OVLT injection of SEA or glutamate was significantly attenuated by pretreatment with an intra-OVLT dose of MK801 (5 microg) or ketamine (10 microg) . The results suggest that glutamatergic pathways in the OVLT region are in pyrogenic fever genesis.

Immunology, 2001 Oct, 104(2), 175 - 84
Porcine dendritic cells generated in vitro: morphological, phenotypic and functional properties; Carrasco CP et al.; Despite the central role that dendritic cells (DC) play in immune regulation and antigen presentation, little is known about porcine DC . In this study, two sources of DC were employed . Bone marrow haematopoietic cell-derived DC (BM-DC) were generated using granulocyte-macrophage colony-stimulating factor (GM-CSF) in the presence or absence of tumour necrosis factor-alpha (TNF-alpha) . Monocyte-derived DC (Momicron-DC) were generated with GM-CSF and interleukin-4 (IL-4) . In both systems, non-adherent cells developed with dendritic morphology, expressing high levels of major histocompatibility complex (MHC) class II . The presence of TNF-alpha increased the BM-DC yield, and enhanced T-cell stimulatory capacity . Both BM-DC and Momicron-DC expressed the pan-myeloid marker SWC3, as well as CD1 and CD80/86, but were also CD14+ and CD16+ . The CD16 molecule was functional, acting as a low-affinity Fc receptor . In contrast, the CD14 on DC appeared to differ functionally from monocyte CD14: attempts to block CD14, in terms of lipopolysaccharide (LPS)-induced procoagulant activity (PCA), failed . The use of TNF-alpha or LPS for DC maturation induced up-regulation of MHC class II and/or CD80/86, but also CD14 . Allogeneic mixed leucocyte reactions and staphylococcal enterotoxin B antigen presentation assays demonstrated that these DC possessed potent T-cell stimulatory capacity . No T helper cell polarization was noted . Both the BM-DC and the Momicron-DC induced a strong interferon-gamma and IL-4 response . Taken together, porcine DC generated in vitro possess certain characteristics relating them to DC from other species including humans, but the continued presence of CD14 and CD16 on mature and immature porcine DC was a notable difference.

Cytokine, 2001 Oct 21, 16(2), 73 - 8
Staphylococcal superantigens induce lymphotactin production by human CD4+ and CD8+ T cells; Tikhonov I et al.; Lymphotactin is a potent chemotactic cytokine (chemokine) that is produced by and also attracts T and natural killer (NK) cells . We are studying whether chemokines that affect mainly T cells might also regulate immune responses by preferentially recruiting individual subsets or by affecting cytokine or other chemokine responses . In order to pursue these questions, we need to learn more about the mechanisms regulating lymphotactin production and the cell types capable of releasing this factor . We used new monoclonal antibodies against human lymphotactin to develop a sensitive antigen-capture enzyme linked immunoabsorbent assay (ELISA) that measures chemokine levels in culture fluids . Using this capture ELISA, we showed that lymphotactin could be produced by CD4+ and CD8+ T cells, but only after T cell-receptor-dependent stimulation using bacterial superantigens and not after treatment by inflammatory cytokines or lipopolysaccharide (LPS) . Our data show that lymphotactin production responds mainly to T cell-receptor signals in CD4+ and CD8+ T cells, and suggests a mechanism whereby this chemokine could help to regulate T cell immune responses .

Tex Heart Inst J, 2001, 28(3), 212 - 4
Purulent and constrictive pericarditis arising from a staphylococcal lumbar infection; Ho JS et al.; A 78-year-old man was admitted to the hospital for evaluation and treatment of anasarca secondary to staphylococcal purulent pericarditis . One month earlier he had undergone a lumbar laminectomy that was complicated postoperatively by a lumbar abscess that was not clinically apparent . The infection subsequently spread to the pericardium . Despite aggressive therapy that included therapeutic pericardiocentesis, drainage of pericardial fluid, and prolonged intravenous antibiotic therapy, the patient returned 2 weeks after discharge from the hospital with complicating constrictive pericarditis . Pericardiectomy was performed, resulting in complete relief of the patient's symptoms.

Res Vet Sci, 2001 Jun, 70(3), 239 - 41
In vitro antibacterial activity of enrofloxacin and ciprofloxacin in combination against Escherichia coli and staphylococcal clinical isolates from dogs; Lautzenhiser SJ et al.; The objective of the study was to determine the in vitro interaction between enrofloxacin and ciprofloxacin against Escherichia coli and staphylococcal isolates from dogs . The microdilution checkerboard assay was used to determine the interaction of the drugs against 50 E . coli and 50 beta-haemolytic staphylococcal clinical isolates . The checkerboard assay revealed that the activity of enrofloxacin and ciprofloxacin was additive against E . coli and staphylococcal clinical isolates . It was concluded that for bacterial species against which ciprofloxacin is more potent than enrofloxacin, the in vivo transformation of enrofloxacin to ciprofloxacin may enhance the efficacy of enrofloxacin, if additivity of the drugs is confirmed in vivo.

Arch Virol, 2001 Aug, 146(8), 1499 - 515
Nitric oxide targets bronchiolar epithelial cells in murine cytomegalovirus-associated disease in lungs that are free of the virus; Tanaka K et al.; At 4 weeks after intraperitoneal (i.p.) injection of 0.2LD50 (50% lethal dose) of murine cytomegalovirus (MCMV) in adult BALB/c mice, productive virus and the viral DNA were detected only in the salivary glands, but not in the lungs . A single i.p . injection of anti-CD3 mAb to these mice provoked pulmonary lesions, which included a thickening of the interstitium and peribronchiolar areas, accompanied with a cellular infiltration in those areas . As a result, half of the mice died . In a histochemical analysis with anti-nitrotyrosine polyclonal Ab, bronchiolar epithelial cells were stained with this Ab, thus demonstrating that peroxynitrite, which was biochemically derived from nitric oxide (NO), injured those cells . Similarly, when T cells of iNOS+/+ mice, which had been infected with MCMV 4 weeks before, were activated by a single injection of anti-CD3 mAb, 37.5% of the mice died . Nitrotyrosine was also detected in the bronchiolar epithelial cells in these mice . In contrast, none of MCMV-infected iNOS-/- mice died after the anti-CD3 injection . No pathological changes were noted in the histological findings of the lungs of those mice . An intranasal injection of bacterial superantigen, staphylococcal enterotoxin B (SEB), demonstrated the same histopathological changes in the lungs and mortality in BALB/c mice as those in mice i.p . injected with anti-CD3 . Therefore, T-cell responsiveness to stimulation with anti-CD3 or a superantigen was presumably modified by MCMV infection . MCMV-associated pneumonitis in the present study was thus mediated not by a direct viral attack but by iNOS-derived NO, which can be induced by the cytokines from the T cells . In addition, it was demonstrated that the NO produced by the cytokine-mediated pathway targeted bronchiolar epithelial cells.

Cancer Immunol Immunother, 2001 Sep, 50(7), 382 - 90
Multiple myeloma cells are killed by syndecan-1-directed superantigen-activated T cells; Ragnarsson L et al.; Multiple myeloma (MM) is an incurable plasma cell/plasmablast malignancy with a great need for innovative treatment strategies . Since experimental immunotherapy with targeted superantigens (SAg) proved to be effective in other haematopoietic tumours, we investigated whether this would also hold true for MM . We used the bacterial SAg Staphylococcus enterotoxin A (SEA), a potent activator of T cell cytotoxicity by means of its binding to particular T cell receptor Vbeta sequences on effector cells and MHC class II molecules on target cells . To eliminate potentially unspecific binding via MHC class II, SEA was point mutated (SEAm) . In a second step SEAm was genetically fused to protein A (PA), resulting in a fusion protein (PA-SEAm) . This fusion protein was used together with four different plasma-cell-specific/associated mAbs to direct T cells towards 10 MM target cell lines . Three of these mAbs were directed against syndecan-1/CD138, known to be highly expressed on MM and plasma cells, but absent on other haematopoietic cells . All MM cell lines proved to be sensitive to SAg-activated T cell killing (15-50% lysis), as measured in a 51Cr-release assay . This effect was clearly mediated via the plasma-cell-reactive antibodies, as control antibodies only conferred a low background lysis . MM therapy based on targeted SAgs could in theory be hampered by dysfunctional T cells in MM patients . However, we show that T cells from MM patients and healthy controls responded equally well to activation by SAg.

Postgrad Med, 2001 Oct, 110(4), 55 - 6, 59-62
Staphylococcal toxic shock syndrome . Suspicion and prevention are keys to control; Issa NC et al.; TSS is still very much with us and can mimic a variety of disorders . Early recognition of the various manifestations of this multisystem disease and careful inspection of possible sites of infection, removal of tampons, and debridement of surgical wounds, along with early aggressive supportive treatment and antibiotic therapy, are critical to prevent complications and ensure recovery . In menstrual TSS, prevention of subsequent relapses is achieved by patient education about proper use of tampons and recognition of early signs of the disease.

Histol Histopathol, 2001 Oct, 16(4), 1149 - 59
Analysis of the in vivo dendritic cell response to the bacterial superantigen staphylococcal enterotoxin B in the mouse spleen; Yoon S et al.; To investigate the in vivo effects of Staphylococcal enterotoxin B (SEB) on dendritic cells (DCs) in the spleen, a single dose of SEB (50 microg/kg) was administered to BALB/c mice by intraperitoneal injection . Afterwards, the mice were sacrificed at 2, 6 and 24 hr, 2, 4, 7 and 15 days, and the spleens were removed . The immunocytochemical characterization of the cells was carried out using various monoclonal antibodies in cryostat-cut sections . The distribution patterns of DCs and their major costimulatory molecules, CD80, CD86 and CD40 in the spleen were identified, and the evidence for maturation of DCs in vivo in response to SEB was obtained . It was found that systemic administration of SEB induced the migration of most of the immature, splenic DCs from the marginal zone to the periarterial lymphatic sheath within 6 hr . This movement paralleled a maturation process, as assessed by upregulation of CD40, CD80 and CD86 expression in the interdigitating dendritic cells (IDCs) . The upregulation of costimulatory molecule expression was conspicuous only in DCs in contrast to other antigen-presenting cells (APCs) such as macrophages and B cells which did not show any significant alterations in their costimulatory molecule expression . We also demonstrated the temporal expression pattern of these costimulatory molecules on the activated DCs . The upregulation of costimulatory molecules on DCs reached a peak level 6 hr after SEB injection, while the increase in number of T cells expressing T cell receptor V138 reached a peak level on day 2 after SEB treatment . In conclusion, we demonstrated the in vivo DC response to SEB in the mouse spleen, especially a potent stimulative effect of SEB on DCs in vivo, a temporal distribution pattern of DCs as well as T cells including TCR Vbeta8+ T cells, and a differential expression pattern of costimulatory molecules on the activated DCs . The results of the present study indicate that DCs are the principal type of APCs which mediate T cell activation by SAg in vivo, and that each costimulatory molecule may have different role in the activation of DCs by SAg . Thus, it is plausible to speculate that DCs play a critical role in the T cell clonal expansion by SAgs and other SAg-induced immune responses in vivo.

Biochem Biophys Res Commun, 2001 Oct 26, 288(2), 454 - 61
Reconstruction of SEA-B7.1 double signals on human hepatocellular carcinoma cells and analysis of its immunological effect; Li Z et al.; Since transfection of established tumors with immunostimulatory genes, such as superantigens (SAg), a family of bacterial and viral proteins with strong immunostimulatory properties, can elicit antitumor immunity, direct transfection of tumors with genes of staphylococcal enterotoxin A (SEA) could probably set up a new way of immunological pathway . In our study, human hepatocellular carcinoma (HCC) cell lines stably transduced with SEA and B7.1/SEA fused genes, HHCCSEA and HHCCBS, were obtained by using the method of retroviral mediated gene transduction . The results showed that human HCC cells could express SEA gene . Although a tiny quantity of expression was detected, a robust immune response was promoted . The cytotoxicity of CTL on HHCCBS was the same as that on HHCCSEA . But the K(m) value of the reaction of the former was lower than that of HHCCSEA . Furthermore, the activity assay of T cells by ELISPOT demonstrated that HHCCBS could elicit more CTL activity than HHCCSEA and HHCCB7.1 . It suggested that the affinity of T cells to HHCCBS was higher and the maxim velocity of reaction could be attained at an early stage of the reaction . Transduced HCC cells were also analyzed for HLA expression, and it was found that a majority of the cells expressed HLA-I molecules but no HLA-DR molecules . After blocking the HLA-I molecules by HLA-I mAb, the cytotoxicity of T lymphocytes dropped remarkably . The results suggested that SEA were mainly presented by HLA-I molecules, and that B7.1 and SEA could have synergistic action at the early stage of the reaction, but the relationship between them in the consequent process needs to be clarified .

J AOAC Int, 2001 Sep-Oct, 84(5), 1587 - 92
Enzyme immunoassay of staphylococcal enterotoxins in dairy products with cleanup and concentration by immunoaffinity column; Lapeyre C et al.; Two different immunoaffinity columns (IACs) were prepared for detection of staphylococcal enterotoxins (SETs) from dairy products . First, a specific IAC for staphylococcal enterotoxin A (SEA), IAC-1, was prepared by coupling monoclonal antibody (mAb) directed against SEA; second, a polyspecific IAC for SEA, staphylococcal enterotoxin B (SEB), staphylococcal enterotoxin C (SECs), and staphylococcal enterotoxin D (SED), IAC-2, was prepared by coupling a mixture of mAbs against SEA, SECs, and SED, and rabbit IgG against SEB . These columns were applied for detection of SETs in dairy products, after extraction, immunoaffinity chromatography, and enzyme immunosorbent assay (EIA) . Overall recoveries from dairy products spiked with 1 ng SEA/25 g averaged 81.2% (range, 76-85%) on IAC-1 . The repeated use of IAC-1 was then determined with good efficiency of 91.5%, in more than 10 runs . On the other hand, a recovery yield of 77% of SETs (SEA, SEB, SEC, and SED) from dairy products spiked with 2.5 ng of each enterotoxin per 25 g, was obtained with IAC-2 . IAC-2 was also successfully subjected to the chromatography of naturally contaminated foods implicated in staphylococcal food poisoning outbreaks . This new extraction-concentration-immunoaffinity-chromatography method (ECIC) is very useful for improving staphylococcal enterotoxin detection and eliminating matrix effect in EIA of dairy products.

J Biol Chem, 2001 Dec 7, 276(49), 46039 - 45 Epub 2001 Oct 11.
Significance of local electrostatic interactions in staphylococcal nuclease studied by site-directed mutagenesis; Leung KW et al.; In this paper, we show that amino acids Glu(73) and Asp(77) of staphylococcal nuclease cooperate unequally with Glu(75) to stabilize its structure located between the C-terminal helix and beta-barrel of the protein . Amino acid substitutions E73G and D77G cause losses of the catalytic efficiency of 24 and 16% and cause thermal stability losses of 22 and 26%, respectively, in comparison with the wild type (WT) protein . However, these changes do not significantly change global and local secondary structures, based on measurements of fluorescence and CD(222 nm) . Furthermore, x-ray diffraction analysis of the E75G protein shows that the overall structure of mutant and WT proteins is similar . However, this mutation does cause a loss of essential hydrogen bonding and charge interactions between Glu(75) and Lys(9), Tyr(93), and His(121) . In experiments using double point mutations, E73G/D77G, E73G/E75G, and E75G/D77G, significant changes are seen in all mutants in comparison with WT protein as measured by fluorescence and CD spectroscopy . The losses of thermal stability are 47, 59, and 58%, for E73G/D77G, E73G/E75G, and E75G/D77G, respectively . The triple mutant, E73G/E75G/D77G, results in fluorescence intensity and CD(222 nm) close to those of the denatured state and in a thermal stability loss of 65% relative to the WT protein . Based on these results, we propose a model in which significant electrostatic interactions result in the formation of a locally stable structure in staphylococcal nuclease.

Infect Immun, 2001 Nov, 69(11), 7190 - 3
Identity of activation molecule 3 on superantigen-stimulated bovine cells is CD26; Lee SU et al.; Most CD8(+) T cells in cultures of bovine mononuclear cells stimulated with staphylococcal enterotoxin C1 develop an unusual phenotype characterized by expression of activation molecule 3 (ACT3) . This superantigen-dependent phenotype may be relevant to immunopathogenesis mediated by certain microbial toxins . The size and N-terminal sequence of immunoprecipitated ACT3 indicate that ACT3 is the bovine orthologue of CD26.

Infect Immun, 2001 Nov, 69(11), 7169 - 72
Comparative analysis of lipopolysaccharide-induced tumor necrosis factor alpha activity in serum and lethality in mice and rabbits pretreated with the staphylococcal superantigen toxic shock syndrome toxin 1; Dinges MM et al.; Host susceptibility to lipopolysaccharide (LPS) is correlated with the levels of circulating tumor necrosis factor alpha (TNF-alpha) that develop in response to circulating LPS . Mice are resistant, relative to rabbits, to the lethal effects of LPS . This study indicates that mice and rabbits are equally sensitive to the lethal effects of circulating TNF-alpha but that mice are more resistant than rabbits to the induction of circulating TNF-alpha by LPS.

Zhongguo Zhong Yao Za Zhi, 1998 Apr, 23(4), 240 - 3, inside back cover
{An antimicrobial experimental study on the traditional drug Anuqitabiao used by the Yi minority nationality}; Jiang Y et al.; The result of experimental study on the antimicrobial effect in vitro of the decoction, water-soluble ingredients and liposoluble ingredients of Anuqitabiao shows that decoction and water-soluble ingredients have certain inhibitory effect on all the tested bacteria, while the liposoluble ingredients do not, indicating the antimicrobial ingredients to be water-soluble . The result of antimicrobial experiment in vivo shows that neither different concentrations nor different separated parts of the decoction have any obvious protective effect on the pneumonococci-infected or staphylococciaureus-infected mice.

Int J Hematol, 2001 Aug, 74(2), 197 - 204
Superantigens and autoantigens may be involved in the pathogenesis of gastric mucosa-associated lymphoid tissue lymphoma; Hashimoto T et al.; To clarify the origin of tumor cells and the possible role of antigens in the pathogenesis of mucosa-associated lymphoid tissue lymphoma (MALTL) of the stomach, we analyzed the DNA sequences of the immunoglobulin (Ig) variable region gene from tumor cells of 4 patients with low-grade and 2 patients with high-grade MALTL associated with Helicobacter pylori infection . There were few somatic mutations in the Ig variable region gene, but intraclonal variations were observed in 2 of the 4 low-grade MALTL cases . In the remaining 2 low-grade MALTL and 1 of the 2 high-grade MALTL cases, somatic mutations and intraclonal variations were evident . In contrast, somatic mutations in the Ig variable region gene were prominent, but intraclonal variation was absent in the other high-grade MALTL cases . The deduced amino acid sequences of the antigen-binding fragments (Fab) from 2 MALTL cases revealed homology with anti-thyroglobulin antibodies, 3 MALTL cases with lupus anti-DNA antibodies, and 1 MALTL case with a rheumatoid factor . Furthermore, the heavy-chain variable region 3 (V(H)3) family genes were used in 5 of the 6 MALTL cases and had conserved amino acid residues for binding to staphylococcal protein A (SpA), a superantigen of B cells . Considering that another superantigen, protein Fv, competes for binding to Fab with SpA and has been shown to play a major role in immune defenses against gut pathogens, SpA and possibly protein Fv may contribute to the development of MALTL . Thus, these observations suggest that most gastric MALTLs arise from memory B cells that are preliminarily activated by superantigens and autoantigens.

Arch Soc Esp Oftalmol, 2001 Sep, 76(9), 545 - 50
{Results in the treatment of acute endophthalmitis by vitrectomy using limbal incision}; Romero Aroca P et al.; PURPOSE: To describe the results obtained in the treatment of acute onset endophthalmitis after cataract surgery in our center using vitrectomy with corneal limbal incisions . METHODS: Study carried out in 12 patients with acute endophthalmitis after cataract surgery in whom a posterior vitrectomy by limbic incisions was performed followed by intravitreal injections of antibiotics (vancomycin + ceftazidime/amikacin) with systemic and topical antibiotic and corticosteroid treatment in the post-operative period . RESULTS: The incidence of endophthalmitis in our center was 0.92% . The most frequently isolated microorganism was Staphylococcus epidermidis (4 cases, 33.33%) . Four cases were negative (33.33%) . A visual acuity equal to or greater than 0.5 at three months was obtained in 6 patients (50%) . Only one case required a second administration of intravitreal injections . One patient suffered vitreal hemorrhaging in the post-operative period, and no other complications were observed in the remaining patients . CONCLUSIONS: We consider that vitrectomy through limbal incisions is a valid alternative for acute onset endophthalmitis treatment.

Am J Cardiol, 2001 Oct 1, 88(7), 750 - 3
Long-term infection rates associated with the pectoral versus abdominal approach to cardioverter- defibrillator implants; Mela T et al.; Infection is an uncommon (0% to 6.7%) but serious complication after implantable cardioverter-defibrillator (ICD) implantation . All ICD primary implants, replacements, or revisions performed at the Massachusetts General Hospital between April 1983 and May 1999 were reviewed . A total of 21 ICD-related infections (1.2%) were identified among 1,700 procedures affecting 1.8% of the 1,170 patients who underwent a primary implant, a generator change, or a revision of their systems . The mean follow-up time was 35 +/- 33 months . Of the 959 patients with long-term follow-up, 19 of the 584 patients (3.2%) with abdominal and 2 of the 375 patients (0.5%) with pectoral systems developed ICD-related infections (p = 0.03) . There was no significant difference between the infection rate among the 959 primary ICD implants and the 447 replacements or system revisions . Only 5 of the patients (24%) had systemic signs of infection, including fever (T>100.5) and elevated white blood count >12,000 . Cultures from the wound revealed staphylococcal species in 16 patients (76%) . Nineteen patients were treated with removal of the entire ICD system in addition to intravenous antibiotics for 2 to 4 weeks . A decrease in the incidence of ICD-related infection has occurred since the advent of transvenous pectoral systems . The main organism responsible for ICD infection is Staphylococcus . The mainstay of ICD infection management consists of complete removal of the entire implanted system.

Cornea, 2001 Oct, 20(7), 720 - 6
Amniotic membrane transplantation in infectious corneal ulcer; Kim JS et al.; PURPOSE: To evaluate the efficacy of amniotic membrane transplantation in the management of treated infectious corneal ulcer in which inflammatory reactions were responsible for corneal damage . METHOD: A prospective study of 21 consecutive eyes (21 patients) was performed . Sufficient antibacterial, antifungal, or antiviral agents were applied to eradicate causative organisms before permanent or temporary amniotic membrane transplantation, or a combination of the two in few patients . The amniotic membrane was soaked in antiinfective agents before transplantation in all cases . RESULTS: After amniotic membrane transplantation, follow-up times ranged from 4 to 28 months (mean, 18 months) . Clinical indications included Staphylococcus species (four cases), Pseudomonas species (five cases), Acanthamoeba species (three cases), fungus (two cases), and herpesvirus (seven cases) . The corneal surface was healed successfully and recurrences of microbial infection were not noted in any case . Visual acuity was improved in cases that were nonscarring or after additional penetrating keratoplasty . CONCLUSION: Amniotic membrane transplantation seems to be a useful adjunctive surgical procedure for the management of infectious corneal ulcer by promoting wound healing and reducing inflammation.

Clin Pediatr (Phila), 2001 Sep, 40(9), 473 - 80
Clinical outcomes of neonatal meningitis in very-low birth-weight infants; Doctor BA et al.; We sought to describe the clinical presentation and consequences of meningitis among 64 very-low-birth-weight (VLBW <1.5 kg) infants who had 67 culture-proven episodes of meningitis over an 18-year period, 1977 through 1995 . Demographic and neonatal descriptors of meningitis and later outcomes were retrospectively examined and neurodevelopmental outcomes of 39 of 45 (87%) meningitis survivors were compared to those of nonmeningitis survivors followed up to 20 months corrected age . Causes of meningitis included coagulase-negative Staphylococcus in 43% of episodes, other gram-positive bacteria in 19%, gram-negative bacteria in 17%, and Candida species in 20% of episodes . Spinal fluid abnormalities were sparse, regardless of etiologic organism . Of 38 nonbloody spinal fluid taps (<1,000 erythrocytes/mm3), 6 had >30 leukocytes/mm3, 5 protein >150 mg/dL%, and 6 glucose <30 mg/dL (1.67 mmol/L) . Only 10 infants (26%) had 1 or more of these spinal fluid abnormalities . Meningitis survivors had a higher rate of major neurologic abnormality (41% vs 11%, p<0.001) and subnormal (<70) Mental Development Index (38% vs 14%, p<0.001) than nonmeningitis survivors . Impairment rates did not differ by etiologic organism . The effect of meningitis on neurologic outcome persisted even after controlling for birth weight, intraventricular hemorrhage, chronic lung disease, and social risk factors (odds ratio 2.27 {95% CI 1.02, 5.05}) . We conclude that despite a sparsity of abnormal spinal fluid findings, culture-proven neonatal meningitis among VLBW infants has a detrimental effect on neurologic outcome, which persists even after controlling for other risk factors.

Arch Pediatr, 2001 Sep, 8 Suppl 4, 665s - 672s
{Development of intensive care for pediatric infectious diseases over the last 20 years}; Huault G; Intensive care and especially pediatric intensive care originated in connection with fatal forms of infectious diseases, some forty years ago . It has come a long way during the last quarter of the past century . Several infectious diseases have disappeared in France during this period such as poliomyelitis, dipteria and tetanus . Many other are receding . Nevertheless problems remain: serious infections by pneumococcus or staphylococcus, maternofoetal infections, whooping cough in very young infant . Now, thanks to liver transplantations and intensive care, the fatal forms of infectious hepatitis can be overcome in more than 50% of the cases . Meanwhile HIV appeared involving intensivists participation . Toxic shock, and especially the purpura fulminans, remains a major difficulty . Great advances in understanding its mechanisms have been made and it seems today like the result of excessive and unsuitable defence reactions . In addition, intensive care takes charge of more and more vulnerable ill persons: immunosuppressed, extreme premature babies, children who had to undergo heavy operations . In those cases, intensive care faces superinfections, nosocomial infections, multi-resistant germs, related in part to the particular working and environmental conditions and an unsuitable use of antibiotics . To conclude, it is necessary to optimize the technical and working conditions in intensive care units, to strictly observe the well established regulations of hygienics and to develop vaccinations.

Int Immunol, 2001 Oct, 13(10), 1291 - 300
In vivo staphylococcal superantigen-driven polyclonal Ig responses in mice: dependence upon CD4(+) cells and human MHC class II; Stohl W et al.; Staphylococcal enterotoxin (SE) B and seven other staphylococcal superantigens (SAg), despite promoting vigorous Ig production in human peripheral blood mononuclear cell cultures, are exceedingly poor at eliciting Ig responses in cultures of spleen cells from C57BL/10J (B10) or C3H/HeJ mice . In contrast, SEB elicits Ig responses in cultures of spleen cells from human MHC class II-transgenic mice . Whereas i.p . administration of SEB (0.2-20 microg) to non-transgenic B10 mice elicits very weak in vivo Ig responses, identical treatment of CD4(+) cell-intact (but not CD4(+) cell-depleted) human MHC class II-transgenic mice elicits dramatic increases in both splenic Ig-secreting cells and serum Ig levels . Over a 2-week period, the SEB-induced in vivo Ig responses peak and then plateau or fall in association with a preferential increase in splenic CD8(+) cells . Nevertheless, in vivo depletion of CD8(+) cells has no sustained effect on SEB-driven Ig responses . Taken together, these observations demonstrate that the effects of SAg on in vivo humoral immune responses are highly CD4(+) cell dependent, are substantially CD8(+) cell independent and can be successfully investigated using human MHC class II-transgenic mice . This model system may be useful in investigating the polyclonally activating effects of microbial products (prototypic environmental insults) on the development of systemic autoimmunity.

Clin Immunol, 2001 Oct, 101(1), 67 - 76
The N-terminal flanking region of the invariant chain peptide augments the immunogenicity of a cryptic "self" epitope from a tumor-associated antigen; Hess AD et al.; The N-terminal flanking region of the invariant chain peptide termed CLIP appears to have superagonistic properties interacting with the T cell receptor and the MHC class II molecule at or near the binding site for the bacterial superantigen Staphylococcal enterotoxin B (SEB) . The present studies explored the hypothesis that the N-terminal segment of CLIP can augment the immunogenicity of cryptic "self" tumor-associated antigens . A chimeric construct of an MHC class II binding peptide from the c-erb oncogene (Her-2/neu) containing the N-terminal flanking region of CLIP elicited potent antitumor activity against a Her-2/neu-positive tumor in a rat model system . Comparatively, the unmodified parent peptide was ineffective . The induction of effective antitumor immunity, however, required presentation of the chimeric peptide construct on irradiated tumor cells or the peptide construct in concert with a Her-2/neu MHC class I-restricted peptide from Her-2/neu . As revealed by adoptive transfer studies, effective protective antitumor immunity in this setting required the CD4 T helper subset . Additionally, in vitro analysis revealed that immunization with the parent peptide resulted in a weak immune response to the unmodified peptide consisting of both type 1 (IL-2, IFN-gamma) and type 2 (IL-4, IL-10) cytokine-producing cells analyzed by RT-PCR (qualitative and quantitative) and by limiting dilution assay . Comparatively, immunization with the chimeric construct elicited a potent immune response to the parent peptide with predominantly type 1 cytokine-producing cells . Taken together, the results suggest that immunization with the chimeric Her-2/neu peptide induced protective antitumor immunity . Associated with this immunization strategy was the enhancement of a type 1 cytokine response .

Am J Kidney Dis . 2001 Oct;38(4):E18.
Shunt nephritis from Propionibacterium acnes in a solitary kidney; Balogun RA et al.; Since its initial description in 1965, immune complex glomerulonephritis associated with ventriculoatrial shunts (VAS) has been reported widely in the literature . The most common incriminating organism is Staphylococcus epidermidis, but less often, an organism generally regarded as nonpathogenic, such as Propionibacterium acnes, has been noted as the cause . Shunt infection usually occurs within a few months after placement or manipulation of the shunt, and shunt nephritis (SN) develops gradually over months to years after . Treatment involves mandatory removal of the shunt and antibiotics; prognosis is variable . We report a case of SN with P acnes that is unusual because of its occurrence in a solitary kidney 6 years after shunt placement, persistently negative blood cultures, and normal complement levels . Percutaneous biopsy of a solitary kidney should be considered if it is expected that the result may guide therapy of progressive renal failure.

Anesth Analg, 2001 Oct, 93(4), 1045 - 9
The continuous femoral nerve block catheter for postoperative analgesia: bacterial colonization, infectious rate and adverse effects; Cuvillon P et al.; We investigated the incidence of bacterial and vascular or neurological complications resulting from femoral nerve catheters used for postoperative analgesia . Patients requiring continuous femoral blockade were consecutively included . Using surgical aseptic procedure, 211 femoral nerve catheters were placed (short-beveled insulated needle, peripheral nerve stimulator) . After 48 h, each catheter was removed and semiquantitative bacteriological cultures were performed on each distal catheter tip . Postoperative analgesia and antibiotics were standardized . All complications during the insertion of the catheters and postoperatively (after 48 h and 6 wk) were noted . Few initial complications with no immediate or delayed complications were noted (20 difficult insertions, 3 impossible injections, 3 ineffective catheters, and 12 vascular punctures) . After 48 h, 208 catheters were analyzed; 57% had positive bacterial colonization (with a single organism in 53%) . The most frequent organisms were Staphylococcus epidermidis (71%), Enterococcus (10%), and Klebsiella (4%) . Neither cellulitis nor abscess occurred . Three transitory bacteremias likely related to the catheter occurred . After 6 wk, no septic complications were noted . One femoral paresthesia, partially recovered 1 yr later, was noted . We conclude that the risk of bacterial complications is small with femoral nerve catheters, although the rate of colonization is frequent . IMPLICATIONS: In this prospective study, continuous femoral nerve catheters were effective for postoperative analgesia but had a frequent rate of bacterial catheter colonization . We found no serious infections after short-term (2-day) infusion . Side effects were few, but one nerve injury occurred.

Avian Dis, 2001 Jul-Sep, 45(3), 745 - 50
Cases of swollen head syndrome in broiler chickens in Greece; Georgiades G et al.; From 50 commercial broiler flocks included in a study concerning respiratory disease, signs of swollen head syndrome (SHS) were shown in eight . Postmortem examination was performed in eight birds showing signs of SHS from each flock . The trachea and head from each bird were collected for laboratory investigation . An enzyme-linked immunosorbent assay (ELISA) was used for the detection of viral and avian mycoplasma antigens in the trachea, and bacteriologic examinations were performed from the infraorbital sinuses of the infected birds . According to the ELISA results, the most frequently detected antigen in the trachea was Mycoplasma synoviae (six flocks, 75%), followed by infectious bronchitis virus (IBV) (five flocks, 62.5%), avian adenovirus (four flocks, 50%), avian reovirus (three flocks, 37.5%), Mycoplasma gallisepticum (one flock, 12.5%), and Newcastle disease virus (NDV) (one flock, 12.5%) . Turkey rhinotracheitis (TRT), infectious laryngotracheitis, and avian influenza viral antigens were not detected . Experimental assays for characterization of NDV and IBV isolates showed that they were strains of low virulence (evidently vaccine strains) . Bacteriologic examinations from the infraorbital sinuses of the affected birds resulted in the isolation of Escherichia coli (seven cases, 87.5%) and Staphylococcus spp . (one case, 12.5%) . It is evident that TRT virus did not play a causal role in SHS in commercial broiler flocks in Greece, but in this condition, other viruses (IBV, NDV), mycoplasmas, or bacteria may be involved, and environmental conditions seem to be essential to the occurrence and severity of the disease.

Eur Cytokine Netw, 2001 Jul-Sep, 12(3), 487 - 500
Cytokine deviation induced by intrathymic injection of staphylococcal enterotoxin B (SEB); Goettelfinger P et al.; We have previously shown that intrathymic (i.t.) injection of staphylococcal enterotoxin B (SEB) to mice induces both T cell clonal deletion and IL-2-dependent anergy . In the present study, we have used a quantitative RT-PCR to demonstrate that i.t . administration of SEB induced a significant decrease in the levels of the IL-2 and IFN-gamma mRNAs in total splenocytes, from day 7 to day 28 post-injection . I.t . SEB injection also induced a significant increase in the levels both of IL-10 and TGF-beta mRNAs on day 7, leading to a significant enhance in the IL-10 + TGF-beta/IL-2 + IFN-gamma mRNA ratio on days 7 and 28 . By contrast, IL-10 and TGF-beta mRNAs were unchanged after intraperitoneal (i.p.) or subcutaneous (s.c.) SEB injections, although both IL-2 and IFN-gamma mRNA levels were decreased . The cytokine mRNA ratio was enhanced on days 7 and 28 after i.p . injection . Interestingly, a cytokine mRNA ratio of a least 10 in favour of IL-10 plus TGF-beta mRNAs was correlated with the hyporesponsive state observed in vitro after i.t . and i.p . injections . Our results clearly demonstrate that i.t . SEB administration induces a switch from Th1-type to Th2-type cytokine expression in the spleen . The deviation from IL-2 plus IFN-gamma towards IL-10 plus TGF-beta expression could be responsible for the immunoregulatory effect exerted upon SEB-reactive T cells, which is characterized by an IL-2-dependent, specific anergy in vitro . Moreover, it highlights the crucial role of the route of SEB injection in the pattern of cytokine expression.

Clin Infect Dis, 2001 Oct 15, 33(8), 1317 - 23 Epub 2001 Sep 17.
Molecular analysis of coagulase-negative Staphylococcus isolates from blood cultures: prevalence of genotypic variation and polyclonal bacteremia; Sharma M et al.; Fifty-seven coagulase-negative Staphylococcus isolates from 22 inpatients who had > or =2 blood cultures that were positive for Staphylococcus within 24 hours were analyzed to determine the frequency of polyclonal bacteremia . Patients were considered to have bacteremia (14 patients) or contamination of sample (8 patients) on the basis of clinical criteria . Nine colonies were randomly selected from each blood culture and genotyped by means of SmaI digestion/pulsed-field gel electrophoresis . Relatedness was determined by calculation of the Dice coefficient of banding-pattern similarity (S(AB)) . Analysis of bacteremic isolates demonstrated the presence of a single species in 35 of 41 blood cultures, 1 related variant in 5 blood cultures (87%-92% S(AB)), and an unrelated strain in 1 blood culture (79% S(AB)) . Analysis of contaminated samples demonstrated the presence of a single strain in 10 of 16 blood cultures and 1-3 variants (28%-97% S(AB)) in the remainder . Genotype diversity was significantly more common in the contaminated samples (P=.036) . Almost all coagulase-negative Staphylococcus bacteremias were monoclonal.

Rev Neurol, 2001 Jun 1-15, 32(11), 1049 - 51
{Spinal epidural abscess secondary to pharyngeal trauma due to food}; Ares B et al.; INTRODUCTION: In recent years the incidence of spinal epidural abscesses has increased . Early diagnosis is very important, since once neurological deficit has become established this is often permanent . The commonest aetiological agent is Staphylococcus . Pain and fever are often the only symptoms before acute neurological deterioration . CASE REPORT: A 76 year old hypersensitive, diabetic man presented complaining of pain and a sensation of warmth in both arms . He had no fever or other symptoms . Some three weeks before admission to hospital a fish bone had become stuck in his pharynx, causing a purulent lesion there . This was treated with antibiotics . Cervical MR showed a mass at the level of C4 C5, compatible with epidural abscess . Following drainage and antibiotic treatment he recovered, with no sequelas . Staphylococcus was isolated from the purulent fluid . CONCLUSIONS: The possibility of epidural infection must be remembered in patients with fever and nerve root pain . Early diagnosis and suitable treatment, combining both surgery and prolonged treatment with antibiotics, are essential to prevent permanent neurological damage.

Microb Pathog, 2001 Oct, 31(4), 185 - 93
Lack of fbe, the gene for a fibrinogen-binding protein from Staphylococcus epidermidis, reduces its adherence to fibrinogen coated surfaces; Pei L et al.; The significance of Fbe, a fibrinogen-binding protein in Staphylococcus epidermidis, was investigated . A fbe mutant was constructed by allelic replacement, where a Gentamicin resistance gene replaced a portion of the A region of fbe . Adherence assay to immobilized fibrinogen on polyethylene surfaces and peripheral venous catheters from patients showed that the fibrinogen binding ability of the mutant was reduced compared to its parental strain . This shows that Fbe is a major factor involved in adherence of S . epidermidis to fibrinogen . No difference was found between the wild-type and mutant in their affinity to immobilized fibronectin .

Comp Immunol Microbiol Infect Dis, 2001 Oct, 24(4), 233 - 46
Chronic fatigue syndrome (CFS) associated with Staphylococcus spp . bacteremia, responsive to potassium arsenite 0.5% in a veterinary surgeon and his coworking wife, handling with CFS animal cases; Tarello W; Chronic fatigue syndrome (CFS) in human patients remain a controversial and perplexing condition with emerging zoonotic aspects . Recent advances in human medicine seem to indicate a bacterial etiology and the condition has already been described in horses, dogs, cats and birds of prey in association with micrococci-like organisms in the blood . To evaluate the possibility of a chronic bacteremia, a veterinary surgeon (the author) and his coworking wife, both diagnosed with CFS and meeting the CDC working case definition, were submitted to rapid blood cultures and fresh blood smears investigations . Blood cultures proved Staph-positive and micrococci-like organisms in the blood were repeatedly observed in the 3-year period preceding the arsenical therapy, during which several medicaments, including antibiotics, proved unsuccessful . Following treatment with a low dosage arsenical drug (potassium arsenite 0.5%, im., 1 ml/12 h, for 10 days) both patients experienced complete remission . At the post-treatment control made 1 month later, micrococci had disappeared from the blood, and the CD4/CD8 ratio was raising.

J Microbiol Immunol Infect, 1999 Mar, 32(1), 40 - 6
Psoas abscess: a 10 year review; Lee YT et al.; Psoas muscle abscesses are rarely encountered yet, and pose diagnostic and therapeutic challenges because of nonspecific clinical presentations . We retrospectively reviewed the medical records of adults with a psoas muscle abscess who were admitted to our hospital from January, 1988 to May, 1998 . Over this ten year period, psoas abscesses were found in 11 cases . Six cases were primary and Staphylococcus spp . was the most commonly isolated . Five cases were a secondary psoas abscess while urological problems were the most common underlying condition . Fever, chills, lower back or flank pain and a palpable mass were the most common manifestations . Only one patient presented the classic triad of fever, flank pain, and limitation of hip movement (a typical psoas sign) . Leukocytosis was the most common laboratory finding . Two of 11 cases presented septic shock . Two patients were admitted to the hospital with the initial diagnosis of psoas abscesses . Computerized tomographic (CT) scans accurately confirmed the clinical diagnosis in 9 of 11 patients . Diagnosis was then established one each by ultrasound (US) and magnetic resonance imaging (MRI), respectively . Four of 11 patients had negative findings initially by US . All patients were managed with drainage and antibiotics, nine were drained surgically, and two were managed with percutaneous drainage under CT guidance . Two patients died . Because of the lack of the classic symptoms and signs in most cases, a high degree of suspicion is important for early diagnosis of psoas abscess . CT scan is the standard technique of diagnosis . The prognosis is generally good with appropriate antibiotic treatment and complete drainage.

J Virol, 2001 Oct, 75(20), 9983 - 5
Differential expression of interleukin-2 and gamma interferon in human immunodeficiency virus disease; Sieg SF et al.; Subnormal T-cell production of interleukin-2 (IL-2) in human immunodeficiency virus (HIV) disease has been described; however, it is not clear whether failure to synthesize IL-2 represents a selective or global defect in T-cell cytokine production . We evaluated the intracellular production of gamma interferon (IFN-gamma) and IL-2 in CD4(+) cells that were stimulated with staphylococcal enterotoxin B or cytomegalovirus antigen . Strikingly, IFN-gamma and IL-2 are differentially regulated in T cells of HIV-infected patients such that the numbers of CD69(+) cells or IFN-gamma-positive cells that make IL-2 are proportionally decreased in CD4(+) T cells from HIV-infected patients . These findings demonstrate a selective defect in IL-2 production and suggest that enumeration of IFN-gamma-producing cells in response to T-cell receptor stimulation, while providing some estimate of antigen-reactive cell frequency, may not reflect or predict "normal" T-cell function in HIV-infected patients.

J Interferon Cytokine Res, 2001 Aug, 21(8), 621 - 9
Type I interferon is the primary regulator of inducible Ly-6C expression on T cells; Schlueter AJ et al.; Ly-6C has been proposed as a marker of memory CD8+ T cells . Reports have indicated that Ly-6C is upregulated after T cell receptor (TCR) stimulation or exposure to proinflammatory cytokines . This study examined the relative roles of proinflammatory cytokines and TCR engagement in Ly-6C induction . In vitro experiments tested the effects of cytokines on Ly-6C expression and confirmed interferon-alpha (IFN-alpha) as a primary cytokine that induces Ly-6C expression on CD4+ and CD8+ T cells . The amount and duration of Ly-6C expression were examined on T cells after in vivo induction of proinflammatory cytokines (CpG oligodeoxynucleotides {ODN}) or TCR activation (staphylococcal enterotoxin B {SEB}) . In vivo, proinflammatory cytokines transiently upregulated Ly-6C on T cells in the absence of TCR stimulation . TCR stimulation by SEB transiently upregulated Ly-6C expression on antigen-specific and antigen-nonspecific T cells but did not cause long-term upregulation of Ly-6C expression in either population . IFN-alpha was confirmed as a primary inducer of Ly-6C in vivo, as CpG ODN were unable to induce Ly-6C expression in IFN-alphaRI(-/-) mice . Thus, inducible Ly-6C expression on CD4+ and CD8+ T cells is largely due to IFN-alpha in the environment and appears not to be directly correlated with the development of T cell memory.

J Endod, 2001 Sep, 27(9), 559 - 62
Long-Term bacterial leakage along obturated roots restored with temporary and adhesive fillings; Barthel CR et al.; The aim of this study was to examine whether obturated roots combined with several adhesive and temporary filling materials can be bypassed by bacteria . Standardized cavities were coronally prepared into 130 straight roots mimicking clinical access cavities . After obturation the roots were assigned to six test and three control groups and coronally sealed with either Clearfil, CoreRestore, IRM, Ketac Fil, or a combination of IRM/wax or Ketac Fil/wax . The roots were then fixed between a top and a bottom chamber each . The top chamber contained soy broth with 108 Staphylococcus epidermidis colony-forming units/ml, and the bottom chamber contained sterile soy broth . For 1 yr the mounts were checked on a regular basis for turbidity in their bottom chambers indicating bacterial growth . After 1 yr only three samples from the CoreRestore group and two samples from the Clearfil group resisted leakage . At termination there was no significant difference in number of leaking samples among the groups . At the beginning of the experiment IRM performed worst . Between months 5 and 10 Clearfil showed the least leaking samples; for some months this was statistically significant compared with IRM or Ketac Fil.

Pediatr Allergy Immunol, 2001 Aug, 12(4), 181 - 7
Ontogeny of T-helper 1 and T-helper 2 cytokine production in childhood; Smart JM et al.; Compared to adults, infants and young children demonstrate differences in their immune response, indicating that there is maturation or change over time and it is probable that this may be reflected in cytokine production . Cytokine responses have been demonstrated to be different in atopic and non-atopic individuals . In this study, we examined T-helper 1 (Th1) (interferon-gamma {IFN-gamma}) and T-helper 2 (Th2) (interleukin {IL}-4, IL-5, and IL-13) cytokine release from atopic and non-atopic children in response to the staphylococcal superantigen, staphylococcal enterotoxin B (SEB) . In non-atopic and atopic children, IFN-gamma, IL-4, and IL-5 release was significantly related to age . Non-atopic children younger than 2 years of age were found to have significantly reduced Th2 (IL-4, IL-5, and IL-13) responses when compared with older, non-atopic children . Atopic children had a reduced IFN-gamma response when compared with non-atopics in early childhood; however, the decreased IFN-gamma response seen in early childhood did not persist after 10 years . These age-related changes in cytokine production provide further support for the concept that cytokine deviations may determine the natural history of atopic disease during early childhood . In addition, the present study indicates the necessity of age-matched controls when examining children for both Th1 (IFN-gamma) and Th2 (IL-4) cytokine release.

Oral Microbiol Immunol, 2001 Oct, 16(5), 257 - 64
Detection and rapid increase of salivary antibodies to Staphylococcus lentus, an indigenous bacterium in rabbit saliva, through a single tonsillar application of bacterial cells; Kanda M et al.; In rabbits, Staphylococcus lentus is one of the major bacteria in saliva and a minor bacteria in jejunum fluids and nasal wash . The presence and induction of naturally occurring antibodies reacting to rabbit indigenous bacteria were studied . In non-immune rabbits, the proportion of anti-S . lentus IgA antibodies to total IgA in the saliva was several times higher than that in the intestinal fluids and the nasal wash . The salivary antibodies were found to have increased 1 week after a single tonsillar application of isolated S . lentus cells but not after a single nasal application or a single intragastric instillation . In addition, the anti-S . lentus antibodies in the saliva highly increased with weekly tonsillar applications of isolated S . lentus but increased only one-fifth with weekly nasal applications of the same cells . These results strongly suggest that the palatine tonsils, which we believe had already been sensitized by S . lentus in the physiological condition, induced naturally occurring antibodies in rabbit saliva.

Transfusion, 2001 Sep, 41(9), 1126 - 9
Presence of Propionibacterium acnes in blood components; Kunishima S et al.; BACKGROUND: Sterility testing, as part of the QC of blood components at the Japanese Red Cross Aichi Blood Center between April 1998 and March 2000, showed that 10 of 5568 tested blood components (0.18%), all of which were RBC concentrates, were contaminated with bacteria . Nine isolates were Propionibacterium acnes and one was Staphylococcus capitis . STUDY DESIGN AND METHODS: To investigate the molecular relatedness of eight available P . acnes isolates, 16S rRNA gene analysis and random amplified polymorphic DNA (RAPD) analysis were performed . RESULTS: DNA sequencing analysis of the 16S rRNA gene showed that five isolates were identified as distinct strains and that three had identical sequences . RAPD analysis in the latter three isolates showed that two exhibited indistinguishable banding patterns that differed from that of the third isolate . CONCLUSION: P . acnes was the most frequent contaminant of blood components, and six of eight isolates were molecularly unrelated . Further studies are necessary to investigate the precise mechanisms of contamination.

Biophys Chem, 2001 Jul 24, 91(3), 273 - 9
Charge transfer during staphylococcal adhesion to TiNOX coatings with different specific resistivity; Poortinga AT et al.; Adhesion of the bacterial strain Staphylococcus epidermidis 3399 to titanium-oxy-nitride (TiNOX) substrata with different specific resistivities was studied in a parallel plate flow chamber, while simultaneously measuring the electric potential of the substrata . During adhesion, bacteria either donated or accepted electrons to the substrata depending on the specific resistivity of the substratum and bacteria that had donated electrons to the substratum adhered more strongly than bacteria that had accepted electrons from the substratum . These results demonstrate that electron transfer plays a role in bacterial adhesion to conducting surfaces, which has hitherto been neglected.

Zh Mikrobiol Epidemiol Immunobiol, 2001 May-Jun, (3), 50 - 3
{Effect of synthetic muramyl dipeptide derivatives on staphylococcal infection in mice}; Grabchenko NI et al.; The work deals with the results of experimental evaluation of the influence of some new modified derivatives of muramyldipeptide (MDP) on the course of staphylococcal infection in mice . The preparations under study were found to produce rapid elimination of bacteria from kidneys and the increase of phagocytic activity of blood macrophages in animals . At the same time MDP and its derivatives stimulated natural killer cells whose activity was inhibited during infection . The dependence between the structure of these compounds and their protective action in staphylococcal infection, as well as the increase of the natural immunity characteristics of the body was followed.

Appl Microbiol Biotechnol, 2001 Aug, 56(3-4), 496 - 501
The effect of osmotic stress on the production of nukacin ISK-1 from Staphylococcus warneri ISK-1; Sashihara T et al.; The effects of several additives on the production of a lantibiotic, nukacin ISK-1, from Staphylococcus warneri ISK-1, in batch fermentation were studied . NaCl, KCl and sorbitol stimulated nukacin ISK-1 production . The addition of 1.4 M NaCl increased nukacin ISK-1 activity 1.5-fold over the control, while cell growth and glucose consumption were inhibited . Nukacin ISK-1 production increased with increasing osmolarity of the medium up to about 3 osmol/kg; however, further increases in osmolarity diminished productivity, irrespective of the kind of additive . Northern blot analysis showed that transcription of the nukacin ISK-1 structural gene (nukA) was activated in the presence of 1.4 M NaC1 . These data indicate that the stimulation effect was due to osmotic stress, which acted, at least in part, at the transcriptional level on the nukA gene.

Can J Anaesth, 2001 Sep, 48(8), 819 - 23
Ketamine isomers suppress superantigen-induced proinflammatory cytokine production in human whole blood; Kawasaki C et al.; PURPOSE: To investigate the efficacy of S(+)-ketamine and R(-)-ketamine on staphylococcal enterotoxin B (SEB)-induced tumour necrosis factor (TNF)-, interleukin (IL)-6, and IL-8 production in human whole blood in vitro . METHODS: After Ethics Committee approval and informed consent, blood samples were obtained from ten healthy volunteers and diluted with five volumes of RPMI 1640 . After adding different doses of ketamine isomers (0-1000 microM), the blood was stimulated with SEB (10 ng x mL(-1)) . After a six-hour incubation period, the plasma TNF- activity was determined by the L929 cell cytotoxic assay and IL-6 and IL-8 concentrations were measured using an enzyme-linked immunoassay . RESULTS: Ketamine isomers significantly suppressed SEB-induced TNF- production at concentrations exceeding 50 microM . Ketamine isomers at concentrations exceeding 100 microM also significantly suppressed SEB-induced IL-6 production . Furthermore, ketamine isomers at concentrations exceeding 500 microM significantly suppressed SEB-induced IL-8 production . There were no significant differences between the suppressive effects of S(+)-ketamine and R(-)-ketamine on SEB-induced proinflammatory cytokine production . CONCLUSION: This study demonstrated that ketamine isomers suppressed SEB-induced TNF-, IL-6, and IL-8 production in human whole blood.

Trans Kans Acad Sci, 1992, 95(1-2), 23 - 8
Glucocorticoid effects on immune cell activation by staphylococcal exotoxins and lipopolysaccharide; Chapes SK et al.; Experiments were conducted to determine the effects of physiologically elevated corticosterone on the activation of macrophages and T cells . These studies find that the elevation of corticosterone does not affect the expression of membrane receptors on macrophages and does not affect the activation of macrophages to produce cytokines . In contrast, elevated corticosterone levels correlate with enhanced T cell proliferation to both mitogens and superantigens.

Saudi Med J, 2000 Mar, 21(3), 270 - 3
Hospital morbidity due to post-operative infections in obstetrics & gynecology; Dhaliwal JK et al.; OBJECTIVE: To study the incidence and risk factors for postoperative infection following cesarean sections and major gynecological surgery . METHODS: Postoperative infection was documented in the specified registers in all patients following cesarean sections and major gynecological surgery from January 1997 to December 1998 . This study was a part of the prospective analysis of hospital-based morbidity and mortality in the Department of Obstetrics and Gynecology . RESULTS: There were a total of 89 cases of postoperative infections amongst 4,032 patients undergoing major operations giving an overall infection rate of 2.2% . The morbidity due to infections was 3.3% in cesarean sections and 0.9% in major gynecological surgery . Abdominal hysterectomies had a higher infection rate than vaginal surgery . The most common causative organisms isolated were Enterococcus, Staphylococcus and Klebsiella species . CONCLUSION: It was found that vaginal flora was a significant source of contamination during surgery, which could be minimised by local sterilisation methods . The high infective morbidity in abdominal hysterectomies needs further analysis of the risk factors . Infection surveillance with a regular review of antibiotic protocols is recommended.

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