Staphylococcus

Przegl Lek, 2003, 60(9), 549 - 52
{Treatment outcome of prosthetic valve endocarditis}; Abramczuk E et al.; BACKGROUND AND AIMS OF THE STUDY: Prosthetic valve endocarditis (PVE) occurred from 1 to 9% of patient after heart valve replacement . The aim of the present study was to assess the long term outcome of the treatment of PVE and to analyze the recurrence of infective endocarditis (IE) . MATERIAL: Hospital mortality, recurrence and late survival were analyzed in 59 patients hospitalized in the Institute of Cardiology for PVE between 1988-95 . There were 26 females and 33 males with age ranging from 17 to 71 years (mean 48) . The onset of PVE was within 60 days from valve replacement in 17 patients and therefore it was considered as early PVE, in 42 patients PVE occurred beyond 60 days from valve replacement was considered as late . METHODS: The criteria of Durack et al . were applied to diagnose PVE . Each patient was classified as having definite PVE, possible PVE, or rejected PVE . The late examinations were performed out-patient in Ist Department of Valvular Heart Disease of Institute of Cardiology . RESULTS: Prosthetic valve replacement in addition to medical treatment was preferred for 34 (58%) of the 59 cases (8 death), whereas 25 (42%) cases were only managed medically (5 death) . The overall hospital mortality rate was 22% (13 patients) . There were 8 (17%) episodes of recurrence with mean interval of 6 months between both episodes . 6 patients were cured, 2 died . Staphylococcal epidermidis infection was present in 63% . Recurrence of IE due to the same microorganism was in 88% of episodes . During follow up (mean 4 years) 5 late deaths occurred . Actual 4 years survival was 69% . CONCLUSIONS: 1 . Mean survival after 4 years was 69%, 2 . Total mortality was 31%, 22% hospital, 9% in late observation (decreased from 8% in first 2 years to 1% mean 4 years after cure, 3 . Recurrence of IE was observed in 17% of episodes, most cases (88%) were caused by the same bacteria as in the primary infection.

Vet Clin North Am Small Anim Pract, 2004 Mar, 34(2), 541 - 55, vii-viii
Medical therapy of otitis externa and otitis media; Morris DO; Medical regimens for the successful treatment of infectious otitis may vary widely depending on the degree of pathologic changes of the external ear canals, the status of the tympanic membranes, and the specific microorganisms involved . Chronic otitis media is especially problematic, because impaction of the bullae can be difficult to resolve and these cases often involve antibiotic-resistant bacterial organisms . In fact, the continuing emergence of resistant Pseudomonas spp and Staphylococcus spp will ensure that the medical therapy of otitis remains a dynamic art for the foreseeable future . This article considers the specific antimicrobial agents available for otic use, their potential toxicities, and guidelines for their use.

J Bacteriol, 2004 Apr, 186(8), 2236 - 9
Metabolic activity of Staphylococcus epidermidis is high during initial and low during late experimental foreign-body infection; Vandecasteele SJ et al.; Foreign-body infection (FBI) is notoriously resistant to eradication by antibiotic treatment . It is hypothesized that reduced bacterial metabolic activity contributes to this resistance . We examined the metabolic activity of Staphylococcus epidermidis in 204 samples recovered during in vitro foreign-body colonization and in 424 samples recovered during in vivo FBI in a rat model . Metabolic activity was measured by determining the amount of 16S rRNA per genome by quantitative PCR . The initial foreign-body-associated growth proved to be a metabolically active process, both in vitro and in vivo . The initial 16S rRNA content was similar to that observed during in vitro exponential-growth phase . However, during late in vivo FBI, a 114-fold (P << 0.0001) decrease in the 16S rRNA content was observed, indicating that there was markedly decreased metabolic activity . This decreased metabolic activity during late FBI can explain at least in part why such infections are so difficult to eradicate with conventional antibiotic treatment.

Ir Med J, 2004 Jan, 97(1), 19 - 20
Staphylococcal diaphyseal subacute osteomyelitis of the ulna in a child: an unusual cause of post-traumatic forearm swelling; Kearns SR et al.; Paediatric subacute osteomyelitis (SAO) presents a diagnostic and therapeutic challenge to clinicians . Typically located in the metaphysis of long bones, diaphyseal SAO of the upper limb is rare . We present the case of a three-year-old girl referred to our fracture clinic as an occult fracture following trauma to her forearm with normal initial radiographs . Follow-up radiographs one week later showed cortical erosion of the distal ulna, while a subsequent MRI scan showed soft tissue swelling with an area of high signal in the distal ulna . A limited biopsy diagnosed staphylococcal subacute osteomyelitis of the ulna . The patient responded to high dose antibiotic therapy and made a full recovery . We present this case to highlight the high index of suspicion required to diagnose and appropriately manage this insidious condition, which may easily be confused with any number of benign and malignant bony lesions and provide a review of the relevant literature.

Int Arch Allergy Immunol, 2004 May, 134(1), 41 - 8 Epub 2004 Mar 25.
Presence of IgE antibodies to bacterial superantigens and increased IL-13-producing T cells in myelitic patients with atopic diathesis; Ochi H et al.; BACKGROUND: Superantigens are considered to exacerbate autoimmune inflammation through the expansion of autoreactive T cells; however, the immune response to bacterial superantigens has not been extensively studied in any type of myelitis . We recently reported the occurrence of a distinct type of myelitis in patients with atopic diathesis, which in a recent nationwide survey was reported to be widespread in Japan . The aim of this study was to investigate the presence of IgE antibodies to bacterial superantigens and the proportion of IL-13- or IL-5-producing CD4+ or CD8+ T cells in patients with myelitis and atopic diathesis . METHODS: Twenty-four myelitic patients with and 12 myelitic patients without hyperIgEemia, 28 patients with multiple sclerosis (MS) and 34 healthy controls were enrolled in this study . IgE antibodies to staphylococcal enterotoxins A (SEA) and B (SEB) in sera were measured using a liquid-phase enzyme immunoassay, and the intracellular production of IL-5 and IL-13 in peripheral blood CD4+ and CD8+ T cells was measured by flow cytometry . RESULTS: The myelitic patients with hyperIgEemia showed significantly higher positive rates of serum SEA/SEB-specific IgE antibodies (41.7 and 62.5%, respectively) than the healthy controls (5.9 and 8.8%), patients with MS (0 and 21.4%) and those with normoIgEemic myelitis (0 and 0%) . Moreover, IL-13-producing CD4+ T cells and CD8+ T cells increased significantly in the myelitic patients with hyperIgEemia compared to the controls, while IL-5-producing CD4+ or CD8+ T cells did not . CONCLUSIONS: The IgE response to staphylococcal superantigens is heightened in myelitic patients with atopic diathesis, which might contribute to increases in IL-13-producing T cells and thus the development of myelitis .

Toxicon, 2004 Mar 15, 43(4), 433 - 8
Immunogenicity of the histidine-to-tyrosine staphylococcal enterotoxin B mutant protein in C3H/HeJ mice; Savransky V et al.; Staphylococcal enterotoxin B (SEB) is a common cause of food poisoning and toxic shock . A safe and effective vaccine is needed to protect against the superantigenic effects of this toxin . We previously constructed and produced an apparently nontoxic SEB mutant having four histidine-to-tyrosine substitutions in positions 12, 32, 105, and 121 . In the present study, we found that this H1.2.3.4 SEB mutant had low toxicity, was able to induce high levels of specific IgG antibodies, and protected mice in both the actinomycin D-primed and intranasal SEB intoxication model systems, despite the absence of detectable specific IgM and IgA antibodies . We propose further development of the H1.2.3.4 recombinant protein as a potential anti-SEB vaccine candidate.

Ophthalmology, 2004 Apr, 111(4), 699 - 705
The epidemiology of acute endophthalmitis after cataract surgery in an Asian population; Wong TY et al.; OBJECTIVE: To describe the incidence, risk factors, and clinical outcome of acute endophthalmitis after cataract extraction in a multiethnic Asian population . DESIGN: Prospective case series . PARTICIPANTS: All patients with cataract extractions performed at the Singapore National Eye Center from 1996 to 2001 . METHODS: Data on patients with acute endophthalmitis cases presenting within 6 weeks after cataract surgery were prospectively collected in a standardized format . MAIN OUTCOME MEASURES: Acute endophthalmitis after cataract surgery . RESULTS: During the study period, 44 803 cataract operations (25 476 phacoemulsification and 19 327 extracapsular cataract extractions) were performed . There were 34 cases of acute endophthalmitis (average annual incidence of 0.076%), 21 of which were culture positive (average annual incidence of 0.040%) . In multivariate analysis, risk of endophthalmitis was associated with phacoemulsification technique (relative risk {RR}, 1.9; 95% confidence interval {CI}, 0.9, 3.9; P = 0.10 for all endophthalmitis cases; RR, 3.1; 95% CI, 1.1, 9.4; P = 0.04 for culture-positive endophthalmitis cases) and the occurrence of intraoperative posterior capsule rupture (RR, 8.0; 95% CI, 3.1, 20.7; P<0.001 for all endophthalmitis cases; RR, 11.0; 95% CI, 3.7, 23.9; P<0.001 for culture-positive endophthalmitis cases) . After a median follow-up of 234 days, half of the eyes achieved a final best-corrected visual acuity of 20/40 . Predictors of this visual acuity included baseline acuity of counting fingers or better, culture-negative endophthalmitis, or infection caused by coagulase-negative Staphylococcus . CONCLUSION: The incidence of acute endophthalmitis after cataract extraction in Singapore is consistent with rates reported elsewhere . The phacoemulsification technique is associated with a higher risk of acute culture-positive endophthalmitis compared with extracapsular cataract extraction . Intraoperative posterior capsule rupture is associated with an 8- to 11-fold higher risk of acute endophthalmitis, suggesting that these eyes should be closely monitored for signs of infection in the immediate postoperative period.

Dev Comp Immunol, 2004 Jun, 28(7-8), 803 - 14
Use of staphylococcal protein A in the analysis of teleost immunoglobulin structural diversity; Bromage ES et al.; Staphylcoccal protein A (SpA) adsorption and sephacryl-S300 filtration were employed to isolate Ig from the sera of six aquaculturally important teleost species; Morone saxatilis (striped bass), Lates calcarifer (barramundi), Oreochromis mossambicus (Mozambique tilapia) and Oreochromis niloticus (Nile tilapia), Salmo salar (Atlantic salmon), and Oncorhynchus mykiss (rainbow trout) . While both gel filtration (S300) and SpA adsorption could purify the 800 kDa tetrameric Ig, SpA demonstrated species-specific variability in the amount retrieved . Virtually 100% of this high molecular weight Ig could be isolated from Mosambique tilapia serum, while 84, 17, 10.7 and 0.5% could be isolated from barramundi, striped bass, Nile tilapia, and Atlantic salmon, respectively . Significant amounts of Ig could not be isolated (<0.1%) from rainbow trout (O . mykiss) serum . All SpA-isolated proteins were approximately 800 kDa in molecular weight and were solely composed of equimolar concentrations of H ( approximately 75 kDa) and L ( approximately 25 kDa) chains . Denaturing, non-reducing electrophoresis demonstrated differential disulfide polymerization (redox forms) consistent with those observed with other teleost species; however, SpA exhibited less affinity for Ig possessing completely polymerized tetramers than the more reduced forms, with the exception of Mossambique tilapia . The existence of three different molecular weight H chains (75, 85, 95 kDa) in Nile tilapia was also observed . Each redox form of Nile tilapia Ig incorporated only one size of H chain.

Anesth Analg, 2004 Apr, 98(4), 1024 - 9, table of contents
The effect of local anesthetics on monocyte mCD14 and human leukocyte antigen-DR expression; Kawasaki T et al.; It has been demonstrated that local anesthetics have several effects on the immune system . Monocytes and macrophages are essential components of the host response to microbial infection; however, the effect of local anesthetics on monocyte surface receptor expression remains unclear . We designed this study to investigate the effects of local anesthetics on monocyte mCD14 and human leukocyte antigen (HLA)-DR expression and lipopolysaccharide (LPS)-induced or staphylococcal enterotoxin B (SEB)-induced tumor necrosis factor (TNF)-alpha production . Blood samples were obtained from 10 healthy volunteers . The effects of local anesthetics on LPS- or SEB-induced TNF-alpha production were determined by using an enzyme-linked immunosorbent assay . After different doses of local anesthetics were added, the blood was stimulated with LPS (10 ng/mL) or SEB (10 micro g/mL) for 4 h . The effects of local anesthetics on monocyte mCD14 and HLA-DR expression were measured by dual monoclonal antibody staining and flow cytometry . Local anesthetics showed no effect on LPS- or SEB-induced TNF-alpha production in human whole blood . Local anesthetics suppressed monocyte HLA-DR expression in a dose-dependent manner (P < 0.05) but had no effect on monocyte mCD14 expression . This study demonstrated that local anesthetics suppress HLA-DR expression on the surface of human monocytes . IMPLICATIONS: Monocyte surface receptors have a crucial role in the host response to microbial infection . We investigated the effects of local anesthetics on monocyte mCD14 and human leukocyte antigen (HLA)-DR expression . Our results show that local anesthetics suppress HLA-DR expression on the surface of human monocytes.

FEMS Immunol Med Microbiol, 2004 Mar 8, 40(2), 173 - 80
Erratum to "Apotransferrin administration prevents growth of Staphylococcus epidermidis in serum of stem cell transplant patients by binding of free iron" . {FEMS Immunol . Med Microbiol . 37 (2003) 45-51}; von Bonsdorff L et al.; We investigated the effect of free, non-transferrin-bound iron occurring in haematological stem cell transplant patients on growth of Staphylococcus epidermidis in serum in vitro, and prevention of bacterial growth by exogenous apotransferrin . S . epidermidis did not grow in normal serum at inoculated bacterial densities up to 10(3) cfu ml(-1) but slow growth could be detected at higher initial inocula . Addition of free iron abolished the growth-inhibitory effect of serum, whereas addition of apotransferrin again restored it . Appearance of free iron and loss of growth inhibition coincided in patient serum samples taken daily during myeloablative therapy . Intravenously administered apotransferrin effectively bound free iron and restored the growth inhibition in patient sera . The results suggest that exogenous apotransferrin might protect stem cell transplant patients against infections by S . epidermidis and possibly other opportunistic pathogens.

Biotechnol Appl Biochem, 2004 Dec, 40(Pt 3), 209 - 28
Biotechnological applications for surface-engineered bacteria; Wernerus H et al.; Display of heterologous proteins on the surface of micro-organisms, enabled by means of recombinant DNA technology, has become an increasingly popular strategy in microbiology, biotechnology and vaccinology . Both Gram-negative and Gram-positive bacteria have been investigated for potential applications . The present review will describe the most commonly used systems for bacterial display, with a focus on the biotechnology applications . Live bacterial vaccine-delivery vehicles have long been investigated through the surface display of foreign antigens and, recently, 'second-generation' vaccine-delivery vehicles have been generated by the addition of mucosal targeting signals, as a means to increase immune responses . Engineered bacteria have also the potential to act as novel microbial biocatalysts with heterologous enzymes immobilized as surface exposed on the bacterial cell surface . They provide the potential for new types of whole-cell diagnostic devices, since single-chain antibodies and other type of tailor-made binding proteins can be displayed on bacteria . Bacteria with increased binding capacity for certain metal ions can be created, and potential environmental or biosensor applications for such recombinant bacteria as biosorbents are being explored . Certain bacteria have also been employed to display various polypeptide libraries for use as devices in in vitro selection applications . Part of the present review has been devoted to a more in-depth description of a promising Gram-positive display system, i.e . Staphylococcus carnosus, and its applications . The review describes the basic principles of the different bacterial display systems and discusses current uses and possible future trends of these emerging technologies.

Biochemistry, 2004 Mar 30, 43(12), 3582 - 9
Probing the folding and unfolding dynamics of secondary and tertiary structures in a three-helix bundle protein; Vu DM et al.; Fast relaxation kinetics studies of the B-domain of staphylococcal protein A were performed to characterize the folding and unfolding of this small three-helix bundle protein . The relaxation kinetics were initiated using a laser-induced temperature jump and probed using time-resolved infrared spectroscopy . The kinetics monitored within the amide I' absorbance of the polypeptide backbone exhibit two distinct kinetics phases with nanosecond and microsecond relaxation times . The fast kinetics relaxation time is close to the diffusion limits placed on protein folding reactions . The fast kinetics phase is dominated by the relaxation of the solvated helix (nu = 1632 cm(-1)), which reports on the fast relaxation of the individual helices . The slow kinetics phase follows the cooperative relaxation of the native helical bundle core that is monitored by both solvated (nu = 1632 cm(-1)) and buried helical IR bands (nu = 1652 cm(-1)) . The folding rates of the slow kinetics phase calculated over an extended temperature range indicate that the core formation of this protein follows a pathway that is energetically downhill . The unfolding rates are much more strongly temperature-dependent indicating an activated process with a large energy barrier . These results provide significant insight into the primary process of protein folding and suggest that fast formation of helices can drive the folding of helical proteins.

Biochemistry, 2004 Mar 30, 43(12), 3336 - 45
Effects of chaotropic and kosmotropic cosolvents on the pressure-induced unfolding and denaturation of proteins: an FT-IR study on staphylococcal nuclease; Herberhold H et al.; FT-IR spectroscopy was used to study the effects of various chaotropic and kosmotropic cosolvents (glycerol, sucrose, sorbitol, K(2)SO(4), CaCl(2), and urea) on the secondary structure and thermodynamic properties upon unfolding and denaturation of staphylococcal nuclease (Snase) . The data show that the different cosolvents have a profound effect on the denaturation pressure and the Gibbs free energy (DeltaG(o)) and volume (DeltaV(o) change of unfolding . Moreover, by analysis of the amide I' infrared bands, conformational changes of the protein upon unfolding in the different cosolvents have been determined . An increase, a reduction, or an independence of the volume change of unfolding is observed, depending on the type of cosolvent, which can at least in part be attributed to the formation of a different unfolded state structure of the protein . The data are compared with the corresponding thermodynamic values of DeltaV(o) for the temperature-induced unfolding process of Snase as obtained by pressure perturbation calorimetry, and significant differences are observed and discussed.

Acta Haematol, 2004, 111(3), 163 - 7
Expansion of TcRalphabeta+CD3+CD4-CD8- (CD4/CD8 double-negative) T lymphocytes in a case of staphylococcal toxic shock syndrome; Carulli G et al.; A 55-year-old woman presented with staphylococcal toxic shock syndrome (TSS) . During the course of the disease a significant lymphocytosis appeared, and a high number of TcRalphabeta+CD3+CD4-CD8- (double-negative, DN) lymphocytes was observed both in bone marrow and in peripheral blood samples . Correction of the altered lymphocyte immunophenotype was observed only 6 weeks after recovery from TSS . The immunophenotype of circulating and bone marrow lymphocytes was also studied during a phase of an aspecific febrile episode observed 2 months after recovery, but no subset of DN lymphocytes was found . A small subset of DN lymphocytes can be found in normal bone marrow, liver, thymus, and skin . These cells show peculiar immune regulatory properties and can increase in certain autoimmune diseases . Our findings may represent a specific effect of lymphocyte stimulation by the staphylococcal exotoxin, which is the effector agent of TSS .

Zh Mikrobiol Epidemiol Immunobiol, 2004 Jan-Feb, (1), 45 - 9
{Immunomodulation activity of new vaccines for pertussis prophylaxis--acellular pertussis vaccine and adsorbed DPT vaccine with acellular component}; Semenova IB et al.; The immunomodulating activity of acellular pertussis vaccine (APV) and adsorbed DPT vaccine with acellular pertussis component (DPTA vaccine) was studied . The study revealed that only large doses of APV, 10 immunizing doses (ID), suppressed humoral and cell-mediated response to sheep red blood cells (SRBC) . 1 ID produced no influence on the formation of antibody producing cells, but increased the development of delayed hypersensitivity (DH) to SRBC . The modulation of cell-mediated immune response, induced by APV, returned to normal after the injection of purified staphylococcal toxoid, used as immunomodulator, in doses of 0.15 BU per mouse and 1.5 BU per mouse . DPTA vaccine containing 1 ID, as well as 10 ID, produced no immunomodulating effect . This was established by the evaluation of humoral response to SRBC in CBA mice and the study of the formation of DH to SRBC in BALB/c mice . As indicated by the total of the presented data, the inclusion of APV into DPTA vaccine enhanced the immunological safety of its pertussis component.

Clin Diagn Lab Immunol, 2004 Mar, 11(2), 239 - 44
Depressed type 1 cytokine synthesis by superantigen-activated CD4+ T cells of women with human papillomavirus-related high-grade squamous intraepithelial lesions; Lee BN et al.; Carcinoma of the cervix is causally related to infection with the human papillomavirus (HPV), and T cells play a pivotal role in the immune response of the host to rid itself of HPV infection . Therefore, we assessed the T-cell function of women with HPV-related cervical neoplasia against a superantigen, Staphylococcus enterotoxin B (SEB) . Each woman provided a cervical brush specimen for HPV DNA testing and Papanicolaou (Pap) smears for the staging of cervical lesions . They also provided a blood specimen for determination of the ability of CD4(+) T and CD8(+) T cells to synthesize Th1 (interleukin-2 {IL-2}, gamma interferon {IFN-gamma}, and tumor necrosis factor alpha {TNF-alpha}) and Th2 (IL-10) cytokines in response to activation with SEB . Compared with control subjects with self-attested negative Pap smears, women with high-grade squamous intraepithelial lesions (HSIL) had significantly lower percentages of activated CD4(+) T cells that produced IL-2 (P = 0.045), IFN-gamma (P = 0.040), and TNF-alpha (P = 0.015) and a significantly lower percentage of activated CD8(+) T cells that produced IL-2 (P < 0.01) . These data indicate that women with HPV-related cervical HSIL show a decrease in Th1 cytokine production by activated CD4(+) T cells and suggested that compromised T-helper functions may negatively impact the function of cytotoxic CD8(+) T cells.

Semin Vasc Surg, 2004 Mar, 17(1), 40 - 4
Management of hemodialysis access infections; Ryan SV et al.; Management of hemodialysis (HD) access infection is one of the most challenging and most common problems faced by surgeons, interventional radiologists, and nephrologists . The goal to eradicate infection is often at odds with the need to maintain access . Patients on HD are immunocompromised and typically have significant comorbid conditions placing them at high risk for the occurrence of access infection . Infection is most common with central-vein catheter access, followed by prosthetic arteriovenous grafts (AVG) and is rare with autogenous fistulas . The diagnosis is usually evident on physical exam, but it is not uncommon for these patients to present with atypical symptoms and lack of clinical findings . Although Staphylococcal species are the most common organism to cause infection, early empiric antimicrobial therapy should also include coverage for Gram-negative organisms . Management of central-vein catheter infection includes removal and delayed replacement or, in patients with mild clinical symptoms, catheter exchange over a guide wire . Our management of AVG infection includes total graft excision when patients present with sepsis or the entire graft is bathed in pus, subtotal graft excision when all of the graft is removed except a small oversewn cuff of prosthetic material on an underlying patent artery, and partial graft excision when only a limited infected portion of the graft is removed and a new graft is rerouted in adjacent sterile tissue to maintain patency of the original graft . This strategy has proven to be highly successful in the management of these complicated cases.

Eur J Biochem, 2004 Mar, 271(5), 962 - 71
Identification of domains on the extrinsic 23 kDa protein possibly involved in electrostatic interaction with the extrinsic 33 kDa protein in spinach photosystem II; Tohri A et al.; To elucidate the domains on the extrinsic 23 kDa protein involved in electrostatic interaction with the extrinsic 33 kDa protein in spinach photosystem II, we modified amino or carboxyl groups of the 23 kDa protein to uncharged methyl ester groups with N-succinimidyl propionate or glycine methyl ester in the presence of a water-soluble carbodiimide, respectively . The N-succinimidyl propionate-modified 23 kDa protein did not bind to the 33 kDa protein associated with PSII membranes, whereas the glycine methyl ester-modified 23 kDa protein completely bound . This indicates that positive charges on the 23 kDa protein are important for electrostatic interaction with the 33 kDa protein associated with the PSII membranes . Mapping of the N-succinimidyl propionate-modified sites of the 23 kDa protein was performed using Staphylococcus V8 protease digestion of the modified protein followed by determination of the mass of the resultant peptide fragments with MALDI-TOF MS . The results showed that six domains (Lys11-Lys14, Lys27-Lys38, Lys40, Lys90-Lys96, Lys143-Lys152, Lys166-Lys174) were modified with N-succinimidyl propionate . In these domains, Lys11, Lys13, Lys33, Lys38, Lys143, Lys166, Lys170 and Lys174 were wholly conserved in the 23 kDa protein from 12 species of higher plants . These positively charged lysyl residues on the 23 kDa protein may be involved in electrostatic interactions with the negatively charged carboxyl groups on the 33 kDa protein, the latter has been suggested to be important for the 23 kDa binding {Bricker, T.M . & Frankel, L.K . (2003) Biochemistry42, 2056-2061}.

Br J Haematol, 2004 Mar, 124(6), 787 - 95
Rituximab for chronic recurring thrombotic thrombocytopenic purpura: a case report and review of the literature; Yomtovian R et al.; Deficiency of von Willebrand factor (VWF) cleaving protease ADAMTS13 has been demonstrated to be the proximate cause of a subset of thrombotic microangiopathic haemolytic anaemias (MAHA) typical for thrombotic thrombocytopenic purpura (TTP) . ADAMTS13 gene mutations cause the hereditary form; acquired deficiency has been attributed to presence of an autoantibody, which may represent a specific subset of MAHA best termed 'autoimmune thrombotic thrombocytopenic purpura' . We describe a patient with relapsing TTP because of ADAMTS13 inhibitors, who failed to achieve sustained remission despite therapies with plasma exchange, steroids, vincristine, staphylococcal protein A and splenectomy . The ADAMTS13 inhibitor titre remained elevated and clinical stability was only maintained by plasma exchange every 2-3 d over a period of 268 d . The patient then received rituximab therapy (eight doses of 375 mg/m2 weekly), during which she required five plasma exchanges in the first 10 d, two exchanges in the next 3 weeks, and none thereafter for 450 d and ongoing . The ADAMTS13 inhibitor titre decreased and enzyme activity increased . We compared this case with that of seven previously reported TTP cases also treated with rituximab; experience suggests that rituximab therapy deserves further investigation for patients with either refractory or relapsing TTP caused by ADAMTS13 inhibitors.

Wien Klin Wochenschr, 2004 Feb 16, 116(3), 98 - 101
Progressive Staphylococcus lugdunensis endocarditis despite antibiotic treatment; Petzsch M et al.; A 68-year old man with fever chills and a diastolic murmur was diagnosed with aortic-valve endocarditis caused by coagulase-negative Staphylococcus lugdunensis . The clinical condition initially improved with antibiotic therapy . On day seven, transoesophageal echocardiography revealed large abscesses extending from the aortic root to the left ventricular wall . Emergency cardiac surgery was performed successfully and a stentless bioprosthetic valve was inserted . S . lugdunensis endocarditis is known for its aggressive clinical course with valve destruction, abscess formation and embolic complications despite appropriate antibiotics . Antibiotic treatment alone is associated with a high mortality rate which can be reduced by early valve replacement.

Proc Natl Acad Sci U S A, 2004 Mar 16, 101(11), 3809 - 14 Epub 2004 Mar 08.
Microsecond folding dynamics of the F13W G29A mutant of the B domain of staphylococcal protein A by laser-induced temperature jump; Dimitriadis G et al.; The small size (58 residues) and simple structure of the B domain of staphylococcal protein A (BdpA) have led to this domain being a paradigm for theoretical studies of folding . Experimental studies of the folding of BdpA have been limited by the rapidity of its folding kinetics . We report the folding kinetics of a fluorescent mutant of BdpA (G29A F13W), named F13W*, using nanosecond laser-induced temperature jump experiments . Automation of the apparatus has permitted large data sets to be acquired that provide excellent signal-to-noise ratio over a wide range of experimental conditions . By measuring the temperature and denaturant dependence of equilibrium and kinetic data for F13W*, we show that thermodynamic modeling of multidimensional equilibrium and kinetic surfaces is a robust method that allows reliable extrapolation of rate constants to regions of the folding landscape not directly accessible experimentally . The results reveal that F13W* is the fastest-folding protein of its size studied to date, with a maximum folding rate constant at 0 M guanidinium chloride and 45 degrees C of 249,000 s(-1) . Assuming the single-exponential kinetics represent barrier-limited folding, these data limit the value for the preexponential factor for folding of this protein to at least approximately 2 x 10(6) s(-1).

Ophthalmologe, 2004 Jul, 101(7), 705 - 9
{Sterility of non-preserved autologous serum drops for treatment of persistent corneal epithelial defects}; Sauer R et al.; PURPOSE: To investigate the sterility of non-preserved autologous serum drops in hospitalized patients with persistent corneal epithelial defects . METHODS: Thirty patients with persistent corneal epithelial defects (aged between 28 and 85, mean 67+/-14 years) were treated with autologous serum drops in five different wards of a university eye hospital between October 2001 and March 2002 . After centrifugation of freshly collected autologous blood, the serum was stored in sterile drop bottles at refrigerator temperature (7 degrees C) and applied undiluted every 1 or 2 h up to 7 days . Using blood, chocolate, endo, and Sabouraud agar media, we assessed the sterility of autologous serum drops on the day before application and on the 4th and 7th days . Examinations performed on the 1st and 4th days included 40 autologous serum drops each and on the 7th day 120 samples (40 serum drops, 40 bottle tops, and 40 bottle bottoms) . The agar media for bacterial examination were read after 48 h of incubation time, and those for fungal examination were read after 21 days . All patients had received topical autologous serum therapy and prophylactic antibiotic drops for at least 7 days . RESULTS: The time period of autologous serum treatment ranged from 7 to 28 (10+/-5) days . On the 1st and 4th days autologous serum drops were sterile . On the 7th day, 3 of 40 (7.5%) samples were contaminated with Staphylococcus epidermidis . Two of these contaminations were due to a combined serum drop and bottle top contamination (5%) . A third case resulted from additional bottle bottom contamination (2.5%) . None of the samples examined showed signs of fungal infection . During application of autologous serum as well as during the follow-up period no patient showed any symptoms of ocular infection . CONCLUSIONS: The application of autologous serum has proven to be a practicable therapy in patients with persistent corneal epithelial defects . When the drops are applied by trained personnel, the absence of contamination can be ensured up to the 4th day . By additional application of prophylactic antibiotic drops, infections may be avoided even if refrigerated non-preserved autologous serum is used up to 7 days.

Phys Rev E Stat Nonlin Soft Matter Phys . 2004 Feb;69(2 Pt 1):021401 . Epub 2004 Feb 19.
Mechanism of protein binding to spherical polyelectrolyte brushes studied in situ using two-photon excitation fluorescence fluctuation spectroscopy; Czeslik C et al.; We used two-photon excitation fluorescence fluctuation spectroscopy with photon counting histogram (PCH) analysis as a new tool to study the binding of globular proteins to colloidal particles in situ . Whereas fluorescence fluctuations are traditionally evaluated by calculating the autocorrelation function (fluorescence correlation spectroscopy), a complementary PCH analysis has been performed in this study which is advantageous when particle concentrations of a multicomponent system are of interest and the particles can be distinguished through particle brightness differences . The binding of two proteins, staphylococcal nuclease (SNase) and bovine serum albumin (BSA), to spherical polyelectrolyte brushes (SPB) was measured as a function of protein concentration and ionic strength of the solution at pH-values where SNase and BSA are positively and negatively charged, respectively . It has been found that SNase and BSA strongly bind to the SPB regardless of the protein charge . When the ionic strength of the solution is raised to 100 mM, the SPB become resistant to both proteins . These findings provide further evidence for a binding mechanism where the proteins are mainly driven to the SPB by the "counterion evaporation" force, while Coulomb interactions play a minor role . The results of this study characterize the potential of SPB as a new class of carrier particles for proteins whose use in biotechnological applications appears to be rewarding.

J Infect Chemother, 2004 Feb, 10(1), 46 - 8
Molecular confirmation of transmission route of Staphylococcus intermedius in mastoid cavity infection from dog saliva; Kikuchi K et al.; We report a case of infection of a mastoid cavity after mastoidectomy had been performed for chronic otitis media with cholesteatoma . The infection was caused by Staphylococcus intermedius after a pet dog had licked the patient's ears . Bacterial strains from the dog's saliva and the otorrhea in the patient were confirmed to be identical by pulsed-field gel electrophoresis analysis . The possibility of an oral transmission route of S . intermedius from pets to humans should be noted.

Arch Pathol Lab Med, 2004 Mar, 128(3), 279 - 81
Bacterial contamination of platelet units: a case report and literature survey with review of upcoming american association of blood banks requirements; Burns KH et al.; The most common transfusion-associated infectious risk in the United States today is bacterial contamination of platelet components . Bacterial contamination is estimated to occur at an incidence of 1:1000 to 1:3000 in platelet units, with severe episodes estimated to occur in about one sixth of contaminated products . Increased awareness and prompt reaction of the medical team can greatly affect the outcome and save a patient's life . The following case history illustrates this issue . A young woman developed chills and rigors while receiving 1 unit of leuko-reduced apheresis platelets for severe thrombocytopenia . The transfusion was stopped, blood cultures were drawn, and the patient developed clinical signs of sepsis . Cultures of both the platelet unit and the patient's blood revealed coagulase-negative Staphylococcus . Microbial susceptibilities in both samples were identical . Pretransfusion blood cultures taken from the patient earlier that day were negative . The platelet unit had been stored for 5 days . We review this case and the literature describing the persistent problem of platelet unit contamination and at the same time highlight the efforts now directed by the American Association of Blood Banks and College of American Pathologists to address this issue . Although there is no uniform approach to dealing with bacterial contamination of platelets, the American Association of Blood Banks and the College of American Pathologists have promulgated new accreditation requirements in an effort to prevent bacterial sepsis associated with platelet transfusion . A new American Association of Blood Banks standard, which will be effective March 1, 2004, requires a combination of strategies both to limit the initial inoculation of bacteria into the blood component and to detect subsequent growth at room temperature (American Association of Blood Banks Association Bulletin #03-12) . The new College of American Pathologists Checklist question, which became effective in December 2003, is a Phase 1 requirement that calls for inspected facilities to have a platelet bacteria detection method in place.

J Perinatol, 2004 Mar, 24(3), 175 - 80
Mortality following blood culture in premature infants: increased with Gram-negative bacteremia and candidemia, but not Gram-positive bacteremia; Benjamin DK et al.; OBJECTIVE: To describe survival following nosocomial bloodstream infections and quantify excess mortality associated with positive blood culture . STUDY DESIGN: Multicenter cohort study of premature infants . RESULTS: First blood culture was negative for 4648/5497 (78%) of the neonates--390/4648 (8%) died prior to discharge . Mortality prior to discharge was 19% in the 161 infants with Gram-negative rod (GNR) bacteremia, 8% in the 854 neonates with coagulase negative staphylococcus (CONS), 6% in the 169 infants infected with other Gram-positive bacteria (GP-o), and 26% in the 115 neonates with candidemia . The excess 7-day mortality was 0% for Gram-positive organisms and 83% for GNR bacteremia and candidemia . Using negative blood culture as referent, GNR {hazard ratio (HR)=2.61} and candidemia (HR=2.27) were associated with increased mortality; CONS (HR=1.08) and GP-o (HR=0.97) were not . CONCLUSIONS: Nosocomial GNR bacteremia and candidemia were associated with increased mortality but Gram-positive bacteremia was not.

Cytotherapy, 2004, 6(1), 23 - 9
Microbial screening of UC blood units by an automated culture system: effect of delayed testing on bacterial detection; Sparrow RL; BACKGROUND: Microbial screening is a mandatory test for banked UC blood (UCB) to comply with the code of good manufacturing practice (GMP) . Cord blood banks (CBBs) are not always closely located to a GMP-licensed microbiology laboratory, resulting in time delays for transport of specimens prior to microbiological testing . This study investigated the influence of >/=24 h delays in initiating automated microbial screening on the detection of bacteria in UCB, by analysis of specimens deliberately spiked with bacteria and the recovery of bacteria from cryopreserved spiked-UCB . MATERIALS AND METHODS: UCB was processed according to standard CBB procedures and spiked with Staphylococcus epidermidis or Escherichia coli {2-2000 colony forming units (CFU)/mL} . Spiked-UCB (0.5 mL) was (1) held at room temperature (RT) and inoculated into pediatric BacT/Alert bottles (bioMerieux) at Days 1, 4 and 7 (delayed inoculation); and (2) inoculated directly (Day 0) into replicate BacT/Alert bottles and held at RT for 1, 4 or 7 days before loading onto the BacT/ALERT system (delayed loading) . Spiked-UCB samples were cryopreserved . Bacterial counts were quantitated on horse blood agar plates . RESULTS: Bacterial growth in UCB spiked with a single bacterium was capable of detection by the BacT/ALERT system . S . epidermidis grew in all conditions of delayed testing (ie . delayed inoculation and delayed loading) . E . coli failed to grow under conditions of delayed inoculation but grew at all time points of delayed loading . S . epidermidis and E . coli were recovered from cryopreserved spiked-UCB . DISCUSSION: Inoculation of culture bottles as soon as possible after sample preparation is preferable . Bacteria can maintain viability in BacT/ALERT bottles inoculated and held at RT for up to 7 days prior to automated culture testing . Bacteria can be successfully recovered from cryopreserved UCB.

J Small Anim Pract, 2004 Feb, 45(2), 113 - 6
Iliopsoas abscess with iliac and femoral vein thrombosis in an adult Siberian husky; Grosslinger K et al.; A nine-year-old, male Siberian husky was presented with fever, decreased appetite and activity, non-weightbearing lameness, and oedematous swelling of the right inguinal and preputial area and the right hindlimb . An abscess within the right iliopsoas muscle, with severe thrombosis of the iliac and femoral vein, was diagnosed by haematology and diagnostic imaging . The abscess and adjoining lymph node were removed surgically through a median coellotomy . The isolated pathogen was Staphylococcus intermedius . Clinical signs resolved completely after surgery . Antimicrobial therapy was continued for four weeks . Within the follow-up period of six months, no recurrence of the clinical signs was detected.

J Pharmacol Sci, 2004 Feb, 94(2), 192 - 6
Cyclooxygenase inhibitors attenuate augmented glutamate release in organum vasculosum laminae terminalis and fever induced by staphylococcal enterotoxin A; Huang WT et al.; Both the hyperthermia and augmented glutamate release in the organum vasculosum laminae terminalis (OVLT) after an intravenous dose (30 ng/kg) of staphylococcal enterotoxin A (SEA) were significantly reduced by pretreatment with intravenous administration of cyclooxygenase inhibitors such as aspirin (1 - 10 mg/kg), sodium salicylate (1 - 10 mg/kg), or diclofenac (10 mg/kg) . Intra-OVLT administration of 50 - 200 microg in 1.0 microl of either aspirin or sodium salicylate 60 min before or 120 min after an intra-OVLT dose (50 microg in 1.0 microl) of glutamate also significantly suppressed the glutamate-induced hyperthermia . These findings suggest that inhibition of cyclooxygenase receptor mechanisms suppresses SEA fever by inhibition of glutamate release in the OVLT of rabbit brain.

Folia Microbiol (Praha), 2003, 48(5), 649 - 53
The effects of magnesium, calcium and EDTA on slime production by Staphylococcus epidermidis strains; Ozerdem Akpolat N et al.; Effect of magnesium, calcium and EDTA on slime production by 15 slime-positive and 13 slime-negative Staphylococcus epidermidis strains isolated from various clinical specimens was determined . The slime production on tryptic soy broth was significantly enhanced after addition of 128 mumol/L Mg2+ . Similarly, the addition of Ca2+ caused a significant increase in slime production of all tested strains when concentration of Ca2+ exceeded 64 mumol/L . In contrast, in the presence of EDTA the slime production by all strains was significantly reduced . Hence Ca2+ and Mg2+ increase slime production of S . epidermidis . This finding is important in the context of the pathogenesis of biomedical implant infections caused by S . epidermidis.

Cochrane Database Syst Rev . 2004;(1):CD001150.
Topical ointment for preventing infection in preterm infants; Conner JM et al.; BACKGROUND: Nosocomial sepsis is a frequent and serious complication of premature infants . The increased susceptibility of ELBW infants to infection has been attributed to less effective immune function compared to mature newborns and the invasive nature of necessary supportive care . Breakdown of the barrier function of the skin may be an additional risk factor for nosocomial sepsis . OBJECTIVES: To assess the effect of prophylactic application of topical ointment on nosocomial sepsis rates and other complications of prematurity in preterm infants . SEARCH STRATEGY: Searches were made of the Cochrane Central Registry of Controlled Trials (CENTRAL, The Cochrane Library, Issue 2, 2003), Ovid DC MEDLINE through June 2003, previous reviews including cross references, abstracts, conference and symposia proceedings, expert informants, and journal hand searching in the English language . SELECTION CRITERIA: Randomized controlled trials which compared the effect of prophylactic application of topical ointment to routine (standard) skin care or as needed topical therapy in preterm infants are included in this review . DATA COLLECTION AND ANALYSIS: Data regarding clinical outcomes including infection {including any bacterial infection, bacterial infection with a known pathogen, coagulase negative staphylococcal infection, fungal infection, and any nosocomial infection (bacterial or fungal)}, patent ductus arteriosus, oxygen requirement at 28 days, chronic lung disease and mortality were excerpted from the reports of the clinical trials by the reviewers . Data analysis was done in accordance with the standards of the Cochrane Neonatal Review Group . MAIN RESULTS: Four randomized controlled trials were identified . All four studies reported improved skin condition in infants treated with prophylactic topical ointment (results not reported here).All four studies reported on the incidence of any nosocomial infection, fungal infection and coagulase negative staphylococcal infection . Infants treated with prophylactic topical ointment are at increased risk of coagulase negative staphylococcal infection (typical relative risk 1.31, 95% CI 1.02, 1.70; typical risk difference 0.04, 95% CI 0.00, 0.08); and any nosocomial infection (typical relative risk 1.20, 95% CI 1.00, 1.43; typical risk difference 0.05, 95% CI 0.00, 0.09) . A trend toward increased risk of any bacterial infection was found in infants treated with prophylactic topical ointment (typical relative risk 1.19, 95% CI 0.97, 1.46; typical risk difference 0.04, 95% CI -0.01, 0.08) . There was no significant difference found in the risk of bacterial infection with a known pathogen, fungal infection, or other complications related to prematurity . REVIEWER'S CONCLUSIONS: Prophylactic application of topical ointment increases the risk of coagulase negative staphylococcal infection and any nosocomial infection . A trend toward increased risk of any bacterial infection was noted in infants prophylactically treated . Topical ointment should not be used routinely in preterm infants.

Diagn Microbiol Infect Dis, 2004 Feb, 48(2), 89 - 95
The effect of systemic antibiotics on the microbiological diagnosis of experimental foreign body infections caused by Staphylococcus epidermidis; Vandecasteele SJ et al.; Qualitative broth culture and quantitative culture on agar were compared with quantitative polymerase chain reaction (PCR) for the diagnosis of foreign body infections (FBI) in a rat model with and without exposure to systemic antibiotics (teicoplanin and rifampin) . The 3 methods had a similar and high yield without antibiotics . Antibiotics decreased the number of CFU/foreign body and increased the number of culture-negative foreign bodies and the variability of the results in quantitative culture . The yield of broth culture remained high under antibiotics although prolonged incubation (2-5 days) was required . The yield of the PCR was equivalent or even superior (for teicoplanin) to the yield of broth culture . Quantitative PCR had a higher yield and lower variability than quantitative culture and was not affected by antibiotics . The simultaneous isolation of RNA from all samples indicated viability of the bacteria . Quantitative PCR seems a promising method for the diagnosis of FBI.

J Otolaryngol, 2003 Dec, 32(6), 388 - 93
Role of microbial toxins in the induction of glucocorticoid receptor beta expression in an explant model of rhinosinusitis; Fakhri S et al.; BACKGROUND: Glucocorticoids (GCs) are the most potent agents currently available for relieving the symptoms of chronic rhinosinusitis . The pathogenesis and molecular basis of GC insensitivity in allergic rhinosinusitis are unknown . Studies done on patients with GC-insensitive asthma demonstrated an overexpression of GC receptor beta (GRbeta), an abnormal splice variant and an endogenous inhibitor of the classic GC receptor alpha . The mechanisms that induce the overexpression of GRbeta remain poorly understood . OBJECTIVE: To study the role of Staphylococcus-derived enterotoxin in inducing GRbeta in a human explant model of rhinosinusitis . METHODS: Nasal tissue was obtained from inferior turbinates of nonatopic and ragweed-sensitive patients . Tissue samples from nonatopic patients were incubated in the presence and absence of superantigen (SAg) of staphylococcal enterotoxin . In addition, tissue samples from ragweed-sensitive patients were incubated with and without ragweed allergen in the presence or absence of SAg . The expression of GRbeta was assessed by immunocytochemistry using a specific polyclonal antibody to GRbeta . RESULTS: SAg increased the expression of GRbeta in both atopic and nonatopic tissue . The highest increase in the expression of GRbeta occurred when atopic nasal tissue was incubated with both ragweed and SAg . CONCLUSION: SAg-induced GRbeta is an important modulator of steroid sensitivity in chronic rhinosinusitis.

J Clin Oncol, 2004 Feb 15, 22(4), 602 - 9
Individualized patient dosing in phase I clinical trials: the role of escalation with overdose control in PNU-214936; Cheng JD et al.; PURPOSE: A patient-specific dose-escalation scheme using a Bayesian model of Escalation with Overdose Control (EWOC) was conducted to establish the maximum tolerated dose (MTD) of PNU-214936 in advanced non-small-cell lung cancer (NSCLC) . PNU-214936 is a murine Fab fragment of the monoclonal antibody 5T4 fused to a mutated superantigen staphylococcal enterotoxin A (SEA) . PATIENTS AND METHODS: Seventy-eight patients with NSCLC were treated with an individualized dose of PNU-214936 calculated using EWOC, based on their anti-SEA antibody level, and given as a 3-hour infusion on 4 consecutive days . RESULTS: Fever (82%; grade 3 to 4, 2.6%) and hypotension (57%; grade 3 to 4, 9%) were the most common toxicities . Eight dose-limiting toxicities occurred, as defined as any grade 4 toxicity occurring within the first 5 days . The MTD was defined as a function of pretreatment anti-SEA antibody level . MTD ranged from 103 ng/kg for patients with anti-SEA concentrations < or = 10 pmol/mL, to 601 ng/kg for patients with anti-SEA concentrations of 91 to 150 pmol/mL . A minor tumor response was demonstrated in five of 66 assessable patients . CONCLUSION: EWOC determined phase I doses of PNU-214936 that were adjusted for patient anti-SEA antibody level, while safeguarding against overdose . Furthermore, the method permitted the construction of a dosing algorithm that would allow patients in subsequent clinical investigations to be treated with a dose of PNU-214936 that is tailored to their specific tolerance for the agent, as reflected by their pretreatment anti-SEA.

Arthritis Rheum, 2004 Feb, 50(2), 589 - 97
Vbeta-restricted T cell adherence to endothelial cells: a mechanism for superantigen-dependent vascular injury; Brogan PA et al.; OBJECTIVE: To investigate the potential for endothelial cells to operate as superantigen-presenting cells for T cells and the potential for such an interaction to cause endothelial cell activation and injury . METHODS: Class II major histocompatibility complex (MHC)-positive human umbilical vein endothelial cells (HUVECs) were cocultured for 4 hours with purified T cells and the superantigens staphylococcal enterotoxin B (SEB) or toxic shock syndrome toxin 1 (TSST-1) . After staining with fluorescence-conjugated monoclonal antibodies, flow cytometric analysis was performed on the HUVECs and T cells to examine V(beta)-restricted T cell adherence to the endothelial cell monolayer, V(beta)-restricted T cell activation (CD69 up-regulation), surface expression of endothelial cell activation markers, and generation of endothelial microparticles (EMPs) . RESULTS: Coculture of purified T cells with class II MHC-positive HUVECs and either TSST-1 or SEB resulted in V(beta)-restricted CD69 up-regulation by CD4 and CD8 cells (V(beta)2 activation for TSST-1; V(beta)3, V(beta)5.1, and V(beta)12 activation for SEB) . Additionally, there was CD4 and CD8 T cell V(beta)-restricted adherence to the HUVEC monolayer at 4 hours . Expression of intercellular adhesion molecule 1, E-selectin, and vascular cell adhesion molecule 1 was up-regulated on the class II MHC-positive HUVECs following exposure to superantigen in the presence of T cells, and there was increased EMP release from activated HUVECs, which occurred earlier and was of greater magnitude than that observed in response to tumor necrosis factor alpha . CONCLUSION: Class II MHC-positive endothelial cells operate as competent superantigen-presenting cells for CD4 and CD8 lymphocytes in vitro . Dual signaling between endothelial cells and T cells results in V(beta)-restricted activation and adherence to endothelial monolayers and endothelial cell activation and release of EMPs expressing inducible cell adhesion molecules . It is proposed that this mechanism could account in part for the vascular injury associated with superantigen-mediated diseases including Kawasaki disease.

Scand J Immunol, 2004 Feb, 59(2), 209 - 19
In vitro study of the long-term effects of post-traumatic splenectomy on cellular immunity; Karakantza M et al.; The purpose of this study was to investigate the effect of splenectomy on cellular immunity . We studied the cellular phenotype and type 1 {interferon-gamma, interleukin-2 (IL-2)} and type 2 (IL-4 and IL-10) cytokine-producing peripheral blood CD4+ and CD8+ T lymphocytes in 22 healthy adults who had undergone post-traumatic splenectomy about 1 to 35 years ago . Splenectomy resulted in a long-term reduction of the percentage of CD4+CD45RA+ cells and a late increase of the percentage and absolute numbers of T-cell receptor gamma/delta cells . Stimulation with Staphylococcal enterotoxin B resulted in normal IL-2 production by CD4+ T cells, indicating that the naive cells were not anergic . Splenectomy also resulted in long-term priming of both CD4+ and CD8+ T cells . During the first 8 years, both type 1 and type 2 CD4+ T cells were primed to varying degrees . About 8 years later, the percentage of primed type 2 CD4+ T cells subsided, but that of type 1 CD4+ T cells, although decreased, remained detectable over a longer period . Priming of CD8+ T cells persisted throughout the study period . The long-term priming of type 1 CD4+ and CD8+ T cells, which may result in partial impairment of T-cell functions, may explain reported defects of immune responses to recall antigens in splenectomized individuals . In addition, changes in the profile of primed CD4+ T cells with time may be clinically relevant to relapses in autoimmune thrombocytopenia after splenectomy.

J Neurosurg, 1971 Feb, 34(2 Pt 1), 145 - 54
A bacteriological study of craniocerebral missile wounds from Vietnam; Carey ME et al.; Bacteriological studies were performed on 45 craniocerebral missile wounds incurred in Vietnam within 2 to 4 hours of occurrence . All missiles had penetrated into the brain . Aerobic and anaerobic cultures were taken of the skin wound, brain, and indriven bone fragments . Forty-four of the skin wounds were contaminated, predominantly with staphylococcus . Only five brain wounds showed bacterial contamination 2 to 4 hours after wounding, indicating that many missile tracks within the brain are initially sterile . Of the patients who had early debridement, 45% had contaminated bone within the brain; possibly up to 75% of all indriven bone chips were sterile . The authors draw the following conclusions . Complete brain debridement with removal of all indriven bone is ideal . Accessible retained bone should be removed by reoperation . Multiple reoperations for an inaccessible retained fragment are inadvisable, however, as fatalities or severe neurologic residua may result . An individual indriven bone chip has a small likelihood of bacterial contamination provided initial debridement was done early . This knowledge may justify an expectant policy in certain individuals harboring an inaccessible retained bone fragment . The retained fragment would be removed only if untoward difficulties develop.

J Neurosurg, 1971 Feb, 34(2 Pt 1), 132 - 41
Early complications following penetrating wounds of the brain; Hagan RE; The experience of an evacuation hospital in Japan in treating 506 consecutive patients from Vietnam with penetrating wounds of the brain is reported with particular reference to early complications . Sixty-eight patients were operated on for still retained intracranial foreign bodies . Thirty-five of the 62 patients with retained intracranial bone fragments had positive microbial cultures of the fragment, which in 63% showed Staphylococcus epidermis . All of the metallic fragments cultured revealed microbial growth . Superficial infections were noted in 32 patients . Superficial plus deep infections were found in eight patients with no retained bone fragments . Eighteen patients had meningitis proven by culture, while an additional 12 patients with CSF sugars of less than 40 mg% were assumed to have meningitis . Twelve patients developed CSF leaks requiring surgery . Twenty-three patients (4.54%) died as a result of their wounds . The neurosurgical treatment recommended for these patients is described.

J Clin Microbiol, 2004 Feb, 42(2), 929 - 32
Treatment of meningitis due to methicillin-resistant Staphylococcus epidermidis with linezolid; Krueger WA et al.; Methicillin-resistant Staphylococcus epidermidis (MRSE) can cause nosocomial meningitis in the presence of prosthetic devices . Vancomycin is the treatment of choice, but its penetration into the cerebrospinal fluid is poor, especially in cases without severe meningeal inflammation . We successfully used linezolid to treat a case of posttraumatic MRSE meningitis with a low-level inflammatory response . Therapeutic effectiveness was documented microbiologically and by the simultaneous measurement of linezolid levels in serum and cerebrospinal fluid.

Protein Expr Purif, 2004 Mar, 34(1), 111 - 7
Development of a large-scale HPLC-based purification for the urease from Staphylococcus leei and determination of subunit structure; Jin M et al.; Coagulase-positive Staphylococcus species, related to but distinct from the genetic homology group containing Staphylococcus cohnii, Staphylococcus xylosus, and Staphylococcus saphrophyticus, were isolated from biopsy material obtained from a cluster of patients in Korea suffering from gastritis . The prototype isolate, Staphylococcus leei, has high urease activity that is similar with respect to a low K(m) value and acid resistance of the urease found in the stomach adapted pathogen, Helicobacter pylori . S . leei is remarkably resistant to lysis and only a small fraction of the cells are broken using sonication, a French press, Niro homogenizer, or a Gaulin mill . In the present report, we describe an efficient cell lysis procedure for S . leei using three passes through a Dynomill with 0.5mm glass beads that results in lysis of more than 95% of the cells . We also developed an efficient and large-scale purification procedure for the S . leei urease using a BioCAD HPLC Workstation using Q-Sepharose, Poros HP2, Sephacryl S-300, and hydroxyapatite chromatography . The urease of S . leei was purified 98-fold to a specific activity of 731U/mg . The urease protein is composed of three subunits, alpha (65kDa), beta (21kDa), and gamma (12kDa), and in situ enzyme assay and molecular sieve chromatography indicate that multiple high molecular weight forms are present, including an apparent pentamer of 1:1:1 alphabetagamma-heterotrimers of 480kDa . This purification procedure was used to purify 16mg of enzyme from 120-liters of cell culture . This improved lysis and purification procedure will make it possible to obtain sufficient quantities of urease for use as an antigen in ELISA assays to carry out studies to determine the incidence and demographic prevalence of gastritis due to S . leei.

J Immunol, 2004 Feb 15, 172(4), 2219 - 24
HIV-infected Langerhans cells preferentially transmit virus to proliferating autologous CD4+ memory T cells located within Langerhans cell-T cell clusters; Sugaya M et al.; Langerhans cells (LC) are likely initial targets for HIV following sexual exposure to virus and provide an efficient means for HIV to gain access to lymph node T cells . The purpose of this study was to examine the nature of the CD4(+) T cell that becomes infected by HIV-infected LC . We infected human LC within tissue explants ex vivo and then, 3 days later, cocultured HIV-infected LC with different subsets of autologous CD4(+) T cells . Using multicolor flow cytometric analyses of LC-CD4(+) T cell cocultures, we documented that HIV-infected LC preferentially infected memory (as compared with naive) CD4(+) T cells . Proliferating and HIV-infected CD4(+) memory T cells were more frequently detected in conjugates of LC and autologous CD4(+) T cells, suggesting that T cells become activated and preferentially get infected through cluster formation with infected LC, rather than getting infected with free virus produced by single HIV-infected LC or T cells . p24(+) Memory CD4(+) T cells proliferated well in the absence of superantigen; by contrast, p24(+) T cells did not divide or divided only once in the presence of staphylococcal enterotoxin B, suggesting that virus production was rapid and induced apoptosis in these cells before significant proliferation could occur . These results highlight that close interactions between dendritic cells, in this case epidermal LC, and T cells are important for optimal HIV replication within specific subsets of CD4(+) T cells . Disrupting cluster formation between LC and memory CD4(+) T cells may be a novel strategy to interfere with sexual transmission of HIV.

Diagn Microbiol Infect Dis, 2004 Jan, 48(1), 17 - 21
Clinical experience with Staphylococcus lugdunensis bacteremia: a retrospective analysis; Ebright JR et al.; We report a retrospective review of all cases of Staphylococcus lugdunensis bacteremia identified during a period of more than 10 years (January 1990 to May 2002) in a large medical center . Twenty-one cases occurred over that period; 20 cases were available for review . In six instances, the organism appeared to be clinically significant and was most commonly related to a vascular line infection . No deaths, secondary suppurative complications, prolonged bacteremia, or prolonged fever appeared to be caused by S . lugdunensis in our experience . Twenty-five percent of our isolates were resistant to multiple antibiotics, including oxacillin.

Clin Microbiol Infect, 2004 Feb, 10(2), 148 - 55
Use of quantitative antibiogram analysis to determine the clonality of coagulase-negative Staphylococcus species from blood culture; Bearson BL et al.; Two phenotypic methods, quantitative antibiogram analysis and colony morphology, were compared to pulsed-field gel electrophoresis (PFGE) for distinguishing the clonality of coagulase-negative Staphylococcus (CNS) species . The results of these three methods were correlated with the patients' clinical findings for 23 episodes in which CNS species were isolated from two blood culture bottles within a 24-h period . Quantitative antibiogram and colony morphology at 24 h correlated with PFGE typing in 21 (91%) and 20 (87%) episodes, respectively . All episodes associated with CNS strains with identical PFGE patterns had quantitative antibiogram similarity coefficients < 10, whereas most episodes associated with strains with different PFGE patterns had quantitative antibiogram similarity coefficients >or= 17 . The CNS isolate pairs were less likely to be associated with infection if the strains had different PFGE types or a quantitative antibiogram similarity coefficient >or= 17 . Clinical microbiology laboratories should consider use of the quantitative antibiogram similarity coefficient to aid clinicians in distinguishing infection-associated CNS blood isolates from contaminants.

Nephron Clin Pract, 2004, 96(1), c15 - 20
Impaired T cell proliferation and zeta chain phosphorylation after stimulation with staphylococcal enterotoxin-B in hemodialysis patients; Eleftheriadis T et al.; BACKGROUND: Patients on regular hemodialysis treatment are in an immunodeficiency state . Several studies have shown defective T cell proliferation after stimulation with various agents . Staphylococcal enterotoxin B (SEB) is a MHC-dependent superantigen that triggers proliferation of a large proportion of T cells . T cell activation after stimulation with SEB parallels normal T cell signal transduction . An important and early event in this transduction pathway is the phosphorylation of the zeta chain . In this study, T cell proliferation and zeta chain phosphorylation after stimulation with SEB were evaluated . METHODS: Peripheral blood mononuclear cells (PBMCs) from 24 patients and 14 healthy individuals were isolated and cultured with or without stimulation with SEB (1 ng/ml) . Cell proliferation was estimated by immunoenzymatic measurement of bromodeoxyuridine uptake . PBMCs from 8 patients and 6 healthy individuals were isolated and pulsed for 2 min with or without SEB (10 microg/ml) . Zeta chain phosphorylation was estimated by immunoprecipitation and immunoblotting with antiphosphotyrosine antibody . RESULTS: Lymphocyte proliferation index after SEB stimulation was lower in hemodialyzed patients . Stimulation of T cells with SEB also resulted in a lower zeta chain phosphorylation in hemodialyzed patients . CONCLUSIONS: Lymphocyte proliferation after MHC-dependent stimulation is impaired in hemodialyzed patients . This proliferation defect is due to impaired zeta chain phosphorylation .

Anal Chem, 2004 Feb 1, 76(3), 684 - 8
Multiplexed toxin analysis using four colors of quantum dot fluororeagents; Goldman ER et al.; Quantum dots (QDs) have the potential to simplify the performance of multiplexed analysis . In this work, we prepared bioinorganic conjugates made with highly luminescent semiconductor nanocrystals (CdSe-ZnS core-shell QDs) and antibodies to perform multiplexed fluoroimmunoassays . Sandwich immunoassays for the detection of cholera toxin, ricin, shiga-like toxin 1, and staphylococcal enterotoxin B were performed simultaneously in single wells of a microtiter plate . Initially the assay performance for the detection of each toxin was examined . We then demonstrated the simultaneous detection of the four toxins from a single sample probed with a mixture of all four QD-antibody reagents . Using a simple linear equation-based algorithm, it was possible to deconvolute the signal from mixed toxin samples, which allowed quantitation of all four toxins simultaneously.

Clin Ther, 2003 Dec, 25(12), 3173 - 89
Cost analysis of switching from i.v . vancomycin to p.o . linezolid for the management of methicillin-resistant Staphylococcus species; McCollum M et al.; BACKGROUND: Infections with methicillin-resistant Staphylococcus species (MRSS) are associated with higher treatment costs than infections with methicillin-sensitive Staphylococcus species in the United States--partly because of an increased length of hospital stay (LOS) . OBJECTIVE: This study used pharmacoeconomic modeling to evaluate the costs and outcomes associated with the use of i.v . vancomycin compared with p.o . linezolid in the treatment of MRSS-infected patients . METHODS: A retrospective chart review was used to determine the number of cases with confirmed or presumed MRSS infections treated with i.v . vancomycin during calendar-year 2000 at the Veterans Affairs Greater Los Angeles Healthcare System inpatient facility . Patients who were eligible for a switch to p.o . linezolid with or without early discharge to home were identified . Cost differences associated with conversion from i.v . to p.o . therapy (compared with continued i.v . therapy) were estimated based on a mean decreased LOS and a decrease in the costs associated with catheter-related adverse events . Rates and costs of catheter-related adverse events were based on estimates from the literature . Sensitivity analyses were performed by variation of the estimated mean LOS decrease in the SD and by variation of the estimates for incidence and costs related to catheter complications . Costs were measured in year 2000 US dollars, and differences were not assessed for statistical significance . RESULTS: Of 177 patients treated with i.v . vancomycin, 103 (58%) were eligible for conversion to p.o . linezolid and 55 (31%) were eligible for early discharge from the hospital with continuation of p.o . therapy . Early discharge was associated with a mean (SD) LOS decrease of 3.3 (2.9) days . Annual mean total cost savings in patients eligible for conversion from i.v . vancomycin to p.o . linezolid with early discharge were $294,750 (range, $35,730-$553,790) . For cases eligible for inpatient conversion from i.v . vancomycin to p.o . linezolid therapy (n=48), the mean total annual cost difference was an increase of $6340 for p.o . linezolid (range, -$12,910 to $11,900) . CONCLUSION: These results--although partly based on estimates from the literature, rather than direct measurements--support the use of p.o . linezolid with or without early discharge as a potential cost-savings alternative for eligible patients treated with a full course of i.v . vancomycin for suspected or confirmed MRSS infection.

Biotechnol Bioeng, 2004 Feb 5, 85(3), 298 - 305
On-line monitoring of important organoleptic methyl-branched aldehydes during batch fermentation of starter culture Staphylococcus xylosus reveal new insight into their production in a model fermentation; de Vos Petersen C et al.; A small fermentor (55 mL) was directly interfaced to a membrane inlet mass spectrometer for continuous on-line monitoring of oxygen and volatile metabolites during batch fermentations of the starter culture Staphylococcus xylosus . Using this technique, we were able to correlate production of the very important flavor compounds 2-methylbutanal, 3-methylbutanal, and 2-methylpropanal with various growth conditions . We found that the aldehydes were present in the culture broth only as transient metabolites . They were produced in the exponential growth phase, reached a maximum concentration when the culture became anaerobic, and then they rapidly disappeared from the culture medium . This general pattern was observed for three different strains of S . xylosus and S . carnosus . Small amounts of inoculum or increased exposure to oxygen were found to favor production of the aldehydes as a result of a longer aerobic growth period . Growing S . xylosus under conditions resembling those in a fermented sausage revealed that NaCl (5%) increased aldehyde production considerably, whereas KNO(3) (0.03%) or NaNO(2) (0.03%) had little effect . A lowering of pH from 7.2 to 6.0 reduced cell density, but had a minor affect on aldehyde production .

Neurosurgery, 2004 Feb, 54(2), 488 - 90; discussion 490-1
Bacterial adhesion to surfactant-modified silicone surfaces; Levy ML et al.; OBJECTIVE: Ventricular shunt infections are a major contributor to morbidity in patients being treated for hydrocephalus . The majority of these infections are from Staphylococcus epidermidis . Prevention of bacterial adhesion to the silicone surface of a ventricular catheter could decrease shunt infections . We studied the effectiveness of a surfactant and/or 2% iodine prewash on preventing bacterial adhesion to Silastic catheter material . METHODS: In a laboratory setting, various concentrations of a surfactant, Poloxamer-188 (P188), and a bactericidal agent, iodine, were compared against a control solution in their ability to prevent bacterial adhesion of S . epidermidis to a silicone surface . Silicone wafers were soaked for 1 hour in the test solution, then inoculated and incubated with S . epidermidis for 24 hours . Bacterial counts were then obtained and compared . RESULTS: The most effective method tested in this study was 20% P188, which allowed only 3.02% bacterial adhesion compared with 22.2% bacterial adhesion in the control (P < 0.001) . P188 at a 10% concentration or 20% mixed with iodine had the next most effective inhibition . Of the germicidal solutions, a 5-ppm solution of iodine was the most effective . The most ineffective method tested was 2 ppm iodine, which allowed 13.2% bacterial adhesion . CONCLUSION: Use of a surfactant and/or a germicidal will provide some protection against bacteria attaching to silicone surfaces before they are surgically implanted . The use of a surfactant soak of 20% P188 or iodine at a concentration of 5 ppm before inoculation with S . epidermidis significantly decreased the bacterial adhesion to silicone wafers . This finding has relevance to clinical practice because it highlights a simple step undertaken before implanting a ventricular catheter that could reduce the adhesion rate of the most common contaminant of these catheters . This step may become an important factor in decreasing infection rates in shunt-dependent patients.

Acta Microbiol Pol, 2003, 52(3), 261 - 9
Synthesis of siderophores by strains of Staphylococcus cohnii isolated from various environments; Szarapinska-Kwaszewska J et al.; Siderophore activity as the feature of microorganisms enabling colonization of human body and the survival in inanimate environment was investigated in 108 strains of Staphylococcus cohnii; S . cohnii ssp . cohnii (50 strains) and S . cohnii ssp . urealyticus (58 strains) . Strains were isolated from people, hospital and non-hospital environment . Highest siderophore activity was noted in strains S . cohnii ssp . urealyticus particularly from the inanimate environments origin . In 86% analyzed strains siderophores of hydroxamate class were detected . Larger amounts of these compounds were synthesized in strains S . cohnii ssp . urealyticus . Strains belonging to both subspecies from human origin showed lower activity of siderophores (total pool) and did not produce hydroxamate class chelators or produced very small amounts of these compounds.

Antimicrob Agents Chemother, 2004 Feb, 48(2), 670 - 2
Penetration of linezolid into the anterior chamber (aqueous humor) of the human eye after intravenous administration; Vazquez EG et al.; We determined the linezolid concentrations in serum samples and aqueous humors (AHs) from 21 patients undergoing cataract extraction . Cataract removal was performed at various times (from 60 to 270 min) after the end of a 30-min infusion of 600 mg of linezolid . Serum samples were obtained 1 h after the end of linezolid administration to determine the maximum concentration of linezolid (C(max)); AHs and a second serum sample were taken simultaneously during the operation, and the concentrations of linezolid in AH (C(AH)) and serum (C(S)) were determined . The mean C(AH) 1 h after linezolid administration was 4.94 micro g/ml, and the mean ratio of C(AH) to C(S) (R = C(AH)/C(S)) was 0.43 . All patients had a C(AH) of >2 micro g/ml, which was higher than the MIC at which 90% of Staphylococcus epidermidis strains are inhibited.

Antimicrob Agents Chemother, 2004 Feb, 48(2), 615 - 8
fexA, a novel Staphylococcus lentus gene encoding resistance to florfenicol and chloramphenicol; Kehrenberg C et al.; The Staphylococcus lentus plasmid pSCFS2 carries a novel florfenicol-chloramphenicol resistance gene, designated fexA, encoding a protein of 475 amino acids with 14 transmembrane domains . The FexA protein differs from all previously known proteins involved in the efflux of chloramphenicol and florfenicol . Induction of fexA expression by chloramphenicol and florfenicol occurs via translational attenuation.

J Dairy Sci, 2003 Dec, 86(12), 3891 - 8
Influence of vacuum level and overmilking on udder health and teat thickness changes in dairy ewes; Peris C et al.; The effects of vacuum level and overmilking on udder health were studied in ewes . Vacuum levels of 36 and 42 kPa were assigned to two groups of 23 Manchega ewes in a crossover study design with two experimental periods of 5 wk for each . Moreover, for each ewe, one teat was overmilked 1.5 to 2 min at all milkings during these 10 wk . The milking machine used had a midlevel milkline and pulsation was fixed at 180 cycles per min and a pulsation ratio of 50:50 . Bacterial exposure of all teats was increased by dipping them in a suspension of Staphylococcus simulans at eight milkings of each period . New intramammary infections (IMI) were not significantly affected by the vacuum level used (18 and 23% of ewes infected, at 36 and 42 kPa, respectively) or application of overmilking (9 and 11% of half udders infected without and with overmilking, respectively) . Likewise, neither factor significantly affected the somatic cell count (SCC) of the milk . Teat thickness changes after milking varied significantly due to the presence of overmilking (-13.6 and -7.4%, in teats not overmilked and overmilked, respectively) but were not affected by vacuum level . At no time were any lesions or variations visibly noted in the teat walls or orifice . So, in this work we were unable to demonstrate that the vacuum and overmilking levels assayed, both used with a pulsation rate of 180 cycles/min, have an important effect on the state of udder health in the short term . Furthermore, it was also observed that, in absence of IMI, the two factors studied did not cause irritation of any kind in the gland that might influence the SCC of the milk.

J Am Anim Hosp Assoc, 2003 Nov-Dec, 39(6), 528 - 32
Septic pericarditis, aortic endarteritis, and osteomyelitis in a dog; Peterson PB et al.; A 7-year-old, female spayed rottweiler was referred with a history of an acute onset of collapse attributable to cardiac tamponade . Thoracic radiographs revealed an enlarged cardiac silhouette compatible with pericardial effusion, sternal osteomyelitis, and an unusual mineralized lesion determined later to be within the aortic wall . The pericardial effusion was a septic exudate secondary to infection with Staphylococcus species and hemorrhage into the pericardium through a mineralized aortic lesion . The case demonstrates the importance of complete evaluation of thoracic radiographs in a patient with cardiac disease and the potential value of cytopathologicalevaluation of pericardial fluid.

Eur J Clin Microbiol Infect Dis, 2004 Feb, 23(2), 98 - 105 Epub 2004 Jan 20.
Immune modulation with a staphylococcal preparation in fibromyalgia/chronic fatigue syndrome: relation between antibody levels and clinical improvement; Zachrisson O et al.; The aims of this study were to evaluate the serological response to treatment with staphylococcal vaccine in fibromyalgia/chronic fatigue syndrome patients and to explore the relationship between serological response and clinical effect . Twenty-eight patients, half of whom served as controls, were recruited from a 6-month randomised trial in which repeated administration of the staphylococcal toxoid vaccine Staphypan Berna (Berna Biotech, Switzerland) was tested against placebo . Antibody status against extracellular toxins/enzymes, cell-wall components, and enterotoxins was evaluated at baseline and at endpoint . The clinical response to treatment was recorded in rating scales . In the group receiving active treatment, significant serological changes were recorded, whereas no significant changes were found in controls . Treatment led to a significantly increased capacity of serum to neutralise alpha-toxin and a significant increase in serum IgG to alpha-toxin and lipase . Furthermore, the increase in these parameters combined paralleled the improvement in clinical outcome . Thus, the greater the serological response, the greater was the clinical effect . In conclusion, this explorative study has shown that repeated administration of the Staphypan Berna vaccine in patients with fibromyalgia/chronic fatigue syndrome causes a serological response to several staphylococcal antigens, particularly to certain extracellular toxins and enzymes . The results further show that this response is related to the clinical outcome of treatment.

Int Immunol, 2004 Feb, 16(2), 315 - 26
Immunologic immaturity, but high IL-4 productivity, of murine neonatal thymic CD4 single-positive T cells in the last stage of maturation; Koyanagi M et al.; To determine the levels of maturation and differentiation of murine CD4 single-positive (SP) T cells, we compared the secondary responses of staphylococcal enterotoxin A (SEA)-induced neonatal thymic, adult thymic and adult splenic CD4 SP T cell blasts prepared from whole or heat-stable antigen(low) CD4 SP T cells . Proliferative responses upon re-stimulation with SEA were strong in adult splenic CD4 SP T cell blasts, but quite weak in neonatal thymic and adult thymic CD4 SP T cell blasts . SEA-induced IL-2 production was weaker in neonatal thymic blasts than in the adult splenic CD4 SP T cell blasts . In contrast, SEA-induced IL-4 production was high in neonatal thymic CD4 SP T cell blasts, and low in adult splenic and thymic CD4 SP T cell blasts . Expression of GATA-3, that directs production of IL-4 in T cells, examined at protein and mRNA levels, was higher in neonatal thymic cells than in adult thymic and splenic cells . These results suggest that neonatal and adult thymic CD4 SP T cells in the final stage of maturation are relatively immature compared with adult splenic CD4 SP T cells . The cytokine production profile of neonatal thymic CD4 SP T cells suggests that they are inclined towards a T(h)2 response.

J Biol Chem, 2004 Apr 30, 279(18), 19239 - 46 Epub 2004 Jan 19.
Suppression of Staphylococcal Enterotoxin B-induced Toxicity by a Nuclear Import Inhibitor; Liu D et al.; Staphylococcal enterotoxin B and related toxins that target T cells have the capacity to elicit systemic inflammation, tissue injury, and death . Genes that encode mediators of inflammation can be globally inhibited by blocking the nuclear import of stress-responsive transcription factors . Here we show that cell-permeant peptides targeting Rch1/importin alpha/karyopherin alpha 2, a nuclear import adaptor protein, are delivered to T cells where they inhibit the staphylococcal enterotoxin B-induced production of inflammatory cytokines ex vivo in cultured primary spleen cells and in vivo . The systemic production of tumor necrosis factor alpha, interferon gamma, and interleukin-6 was attenuated in mice either by a cell-permeant cyclized form of SN50 peptide or by a transgene whose product suppresses the nuclear import of transcription factor nuclear factor kappa B in T cells . The extent of liver apoptosis and hemorrhagic necrosis was also reduced, which correlated with significantly decreased mortality rates . These findings highlight nuclear import inhibitors as a potentially useful countermeasure for staphylococcal enterotoxin B and other toxins that trigger harmful systemic inflammatory responses.

Neurosci Lett, 2004 Jan 23, 355(1-2), 33 - 6
Antipyretic effect of acetaminophen by inhibition of glutamate release after staphylococcal enterotoxin A fever in rabbits; Huang WT et al.; The present study was designed to determine whether the inhibition of glutamate release in organum vasculosum laminae terminalis (OVLT) of rabbit brain by acetaminophen might be protective in a whole-animal model of staphylococcal enterotoxin A (SEA) fever . Unanesthetized rabbits were administered intravenously with SEA, and both colonic temperature (Tco) and glutamate release in OVLT were measured simultaneously . The glutamate release in OVLT was measured with a microdialysis probe previously implanted . Both the Tco and glutamate release in OVLT were simultaneously increased following intravenous administration of SEA . The SEA-induced rise in both the Tco and the levels of glutamate release in OVLT were suppressed by pretreatment with intravenous injection of acetaminophen (1, 5 or 10 mg/kg) . Furthermore, treatment of OVLT with acetaminophen (50-150 microg) attenuated the fever-like hyperthermia induced by intra-OVLT injection of glutamate . Our results show acetaminophen may reduce glutamate release in OVLT and result in antipyresis.

J Am Acad Dermatol, 2004 Feb, 50(2), 266 - 72
Temperature regulates bacterial protein production: possible role in rosacea; Dahl MV et al.; Facial skin temperature is higher for patients with rosacea . Papules and pustules might arise because bacteria behave differently at these warmer temperatures . We sought to: (1) compare bacteria from facial skin of patients with rosacea with that of control subjects; and (2) grow these bacteria at 30 degrees C and 37 degrees C to compare growth curves and secreted proteins . Bacteria isolated from pustules/skin surfaces of patients with rosacea and skin surfaces of control subjects were identified and cultured at 37 degrees C and 30 degrees C . Secreted proteins were separated by electrophoresis . We found that Staphylococcus epidermidis isolated from patients with rosacea was consistently beta-hemolytic, whereas that from control subjects were nonhemolytic . Bacteria from patients with rosacea grew at the same rate and to the same stationary phase whether cultured at 37 degrees C or 30 degrees C . Isolates from patients with rosacea secreted more proteins, and generally more of each protein at 37 degrees C compared with 30 degrees C . In conclusion, bacteria isolated from patients with rosacea secrete different proteins and different amounts of protein at different temperatures.

J Pediatr, 2004 Jan, 144(1), 93 - 9
Autosomal recessive hyperimmunoglobulin E syndrome: a distinct disease entity; Renner ED et al.; OBJECTIVE: The autosomal-dominant form of the hyperimmunoglobulin E syndrome (AD-HIES) has been described as a multisystem disorder including immune, skeletal, and dental abnormalities . Variants of AD-HIES are known but not well defined . METHODS: We evaluated 13 human immunodeficiency virus-seronegative patients from six consanguineous families with an autosomal-recessive form of hyperimmunoglobulin E syndrome (AR-HIES) and 68 of their relatives . RESULTS: Persons affected with AR-HIES presented with the classical immunologic findings of hyperimmunoglobulin E syndrome, including recurrent staphylococcal infections of the skin and respiratory tract, eczema, elevated serum immunoglobulin E, and hypereosinophilia . In addition, severe recurrent fungal and viral infections with molluscum contagiosum, herpes zoster, and herpes simplex were noted . Autoimmunity was seen in two patients . Central nervous system sequelae, including hemiplegia, ischemic infarction, and subarachnoid hemorrhages, were common and contributed to high mortality . Notably, patients with AR-HIES did not have skeletal or dental abnormalities and did not develop pneumatoceles, as seen in AD-HIES . In lymphocyte proliferation assays, patients' cells responded poorly to mitogens and failed to proliferate in response to antigens, despite the presence of normal numbers of lymphocyte subpopulations . CONCLUSION: The autosomal-recessive form of hyperimmunoglobulin E syndrome is a primary immunodeficiency with elevated immunoglobulin E, eosinophilia, vasculitis, autoimmunity, central nervous system symptoms, and high mortality . AR-HIES lacks several of the key findings of AD-HIES and therefore represents a different, previously unrecognized disease entity.

J Infect, 2004 Feb, 48(2), 168 - 74
Serological evidence of Helicobacter pylori infection in UK South Asian and European populations: implications for gastric cancer and coronary heart disease; Fischbacher CM et al.; OBJECTIVES: To describe the prevalence of serological evidence of infection with Helicobacter pylori among people of South Asian and European ethnic origins and to assess its association with prevalent coronary heart disease (CHD) . METHODS: We used a quantitative method to compare IgG antibodies to H . pylori in a population sample of 300 South Asians and 302 Europeans in Newcastle upon Tyne, UK . RESULTS: For men and women, respectively, H . pylori IgG (95% confidence interval) was 16.7 microg/ml (13.9, 20.2) and 11.3 (9.4, 13.5) among Europeans and 11.6 (9.8, 13.7) and 14.3 (12.1, 16.9) among South Asians . Levels were higher in older participants and in those of lower socioeconomic status . The ratio of geometric mean IgG, (95% confidence interval) adjusted for age, sex and socioeconomic status, in those with and without CHD was 1.02 (0.49, 2.11) among Europeans and 1.79 (1.01, 3.17) among South Asians . Antibodies against staphylococcal enterotoxins A and B were higher among South Asians than Europeans . CONCLUSIONS: The prevalence of H . pylori infection among UK South Asians does not reflect that of their countries of origin, nor their lower prevalence of gastric cancer . The association with CHD in South Asians requires corroboration in other studies.

Br J Haematol, 2004 Feb, 124(3), 315 - 24
Signalling molecules and cytokine production in T cells of multiple myeloma-increased abnormalities with advancing stage; Mozaffari F et al.; T-cell immune dysfunction in patients with malignant tumours has been attributed to the altered expression of components of the T-cell receptor (TCR)/CD3 complex and their associated intracellular protein tyrosine kinases . In this study, four-colour flow cytometry was applied to study the surface bound molecules TCRalphabeta, CD28, CD152 and CD154 involved in T-cell signalling and the signal transduction molecules CD3zeta, p56lck, p59fyn, ZAP-70 and phosphatidyl-inositol-3 kinase (PI3-k) as well as the intracellular cytokines interferon-gamma (IFN-gamma), interleukin (IL)-4 and IL-2 as a functional read-out of non-stimulated and superantigen (staphylococcus enterotoxin B)-stimulated blood T cells of multiple myeloma (MM) patients at different stages of the disease . Multiple abnormalities were demonstrated in the CD4 and CD8 populations, both under non-stimulated and superantigen-stimulated conditions . There was a marked reduction, particular in advanced stage MM, in the proportion of CD4 and CD8 cells expressing CD28, CD152, CD3zeta, p56lck, ZAP-70 and PI3-k . The level of intracellular T-cell cytokines (IFN-gamma, IL-2 and IL-4) was normal or increased in non-stimulated cells but activation-induced cytokine production was impaired . These results illustrated profound and multiple T-cell signalling defects, from the surface and down-stream, consistent with involvement of a master T-cell function, especially in advanced stage MM . These data should be taken into consideration when developing immune-based therapeutic approaches and when applying new emerging technologies that aim to restore T-cell functions.

Arch Microbiol, 2004 Mar, 181(3), 250 - 3 Epub 2004 Jan 09.
Specificity grouping of the accessory gene regulator quorum-sensing system of Staphylococcus epidermidis is linked to infection; Carmody AB et al.; Staphylococcus epidermidis represents the most frequent pathogen involved in nosocomial infections and infections of indwelling medical devices . The strain-to-strain variation of the gene encoding the quorum-sensing pheromone of S . epidermidis as well as the correlation between specificity groups and origin from infection were determined . The pro-pheromone gene was highly conserved and showed infrequent, non-synonymous, single-nucleotide polymorphisms that led to conservative amino acid exchanges only . Importantly, one specificity group was significantly more frequent among strains isolated from infection . The finding that quorum-sensing specificity groups are linked to infection demonstrates the relevance of quorum-sensing for virulence in this critical human pathogen and contributes to the scientific basis needed for the development of quorum-sensing-targeting drugs.

Proteins, 2004 Jan 1, 54(1), 8 - 12
Sequence-based study of two related proteins with different folding behaviors; Favrin G et al.; Z(SPA-1) is an engineered protein that binds to its parent, the three-helix-bundle Z domain of staphylococcal protein A . Uncomplexed Z(SPA-1) shows a reduced helix content and a melting behavior that is less cooperative, compared with the wild-type Z domain . Here we show that the difference in folding behavior between these two sequences can be partly understood in terms of an off-lattice model with 5-6 atoms per amino acid and a minimalistic potential, in which folding is driven by backbone hydrogen bonding and effective hydrophobic attraction .

Eur J Haematol, 2003 Dec, 71(6), 464 - 5
Necrotising dermatitis in refractory acute myeloid leukaemia; D'Apollo N et al.; Severe cutaneous infections in leukaemic patients are difficult to treat and can rapidly become fatal . We report on a case of essential thrombocythemia evolved to a myelodysplastic syndrome and finally, to an overt myeloid leukaemia, refractory to chemotherapy . In the presence of a marked neutropenia, the patients developed a wide Staphylococcus epidermidis necrotising dermatitis . The diagnosis was made possible only by a skin biopsy culture and the antibiotic treatment, based on antimicrobial susceptibility tests, rapidly resolved the infection . In neutropenic patients, appropriate laboratory tests and treatment, can lead to recovery of life-threatening infections.

Vet Radiol Ultrasound, 2003 Nov-Dec, 44(6), 648 - 51
Inflammatory polyp in the middle ear with secondary suppurative meningoencephalitis in a cat; Cook LB et al.; A 15-month-old male Maine Coon Cat presented with persistent auricular discharge and progressive head tilt, ataxia, and loss of blink on the right side . Using computed tomography a hyperattenuating, contrast-enhancing material within a thickened right tympanic bulla and contrast enhancement of the adjacent cerebellum were identified . Marked suppurative inflammation was identified on cerebrospinal fluid analysis with no growth on bacterial culture . Ventral bulla osteotomy was performed to remove a soft tissue mass, and an inflammatory polyp with chronic severe suppurative inflammation was confirmed using histology . Staphylococcus auricularis was grown on aerobic culture and Fusobacterium necrophorum and Peptostreptococcus anaerobius were grown on anaerobic culture . The cat was treated for 10 weeks with amoxicillin/clavulinic acid and metronidazole . Dramatic improvement in body weight, appetite, energy level, balance, and resolution of right-sided facial paralysis were noted, but the cat retained a head tilt.

Med Dosw Mikrobiol, 2003, 55(3), 225 - 9
{Selected features determine pathogenicity of Staphylococcus haemolyticus}; Bartoszewicz M et al.; The study have been done on S . haemolyticus strains isolated from patients hospitalized an Surgical Unit . Aim of the study was to determine pathogenic traits of S . haemolyticus: slime producing, adhesion to biomaterials, antibiotics susceptibility and the profiles of surface proteins . Among 44 S . haemolyticus strains, in the test-tube method, there have been 38% labeled as slime producing and 62% as non-producing . In the plate method at 48% slime production was noticed, while 52% strains did not produce slime . It is quite significant that all CNS strains which have an ability to produce mucus, that was proved by means of two methods (test-tube and plate), show a high level of TTC's reduction to formazan . The analysis of resistance to antibiotics in relation to slime production demonstrated more frequent antibiotic resistance of the slime-producing strains.

Infect Control Hosp Epidemiol, 2003 Dec, 24(12), 961 - 3
Clinical experience with minocycline and rifampin-impregnated central venous catheters in bone marrow transplantation recipients: efficacy and low risk of developing staphylococcal resistance; Chatzinikolaou I et al.; In this retrospective evaluation of the 4-year clinical use of minocycline and rifampin-impregnated catheters in bone marrow transplantation (BMT) patients, we report low risk of development of staphylococcal resistance to the antibiotics coating the catheters and efficacy in preventing primary staphylococcal bloodstream infections.

Eur J Ophthalmol, 2003 Nov-Dec, 13(9-10), 773 - 8
The effect of intraoperative antibiotics in irrigating solutions on aqueous humor contamination and endophthalmitis after phacoemulsification surgery; Sobaci G et al.; PURPOSE: To determine the efficacy of intraoperative antibiotic use in irrigating solutions on aqueous humor contamination during phacoemulsification surgery and to evaluate the corresponding risk of postoperative endophthalmitis . METHODS: 644 eyes of 640 patients who underwent phacoemulsification surgery with foldable intraocular lens (IOL) implantation were included in this study . Preoperative conjunctival smears were taken and eyes were allocated to receive irrigating infusion fluid containing either balanced salt solution (BSS)-only (group 1; 322 eyes) or BSS with antibiotics (vancomycin and gentamycin) during surgery (group 2; 322 eyes) . Bacterial contamination rates of aqueous humor samples taken in the beginning and at the end of operation were compared . Predictive factors for the development of postoperative endophthalmitis were determined by clinical and microbiologic analyses . RESULTS: The rates of culture-positivity were similar between group 1 and group 2 for both preoperative conjunctival smears and aqueous samples (p > 0.05) . Aqueous samples taken at the end of operation were found to be contaminated in 68 (21.1%) eyes in group 1 and 22 (6.8%) eyes in group 2, and the difference was significant (p = 0.0001; OR = 3.65 (2.1-6.0)) . Capsular rupture was associated with higher rate of contamination in both groups (p = 0.0001; OR = 7.7 and p = 0.0001; OR = 8.1) . Two eyes in the BSS-only group developed postoperative endophthalmitis and these cases had posterior capsular rupture during the surgery and culture-positivity for staphylococcus epidermidis throughout the study . CONCLUSIONS: Intraoperative antibiotic irrigation decreases aqueous humor contamination during phacoemulsification . Further studies are warranted to determine the interrelationship between aqueous humor contamination and endophthalmitis in eyes with posterior capsular rupture.

World J Gastroenterol, 2004 Jan, 10(1), 53 - 7
Superantigen-SEA gene modified tumor vaccine for hepatocellular carcinoma: an in vitro study; Lu SY et al.; AIM: To construct an eukaryotic superantigen gene expression vector containing the recombinant gene of SEA and CD80 molecule transmembrane region (CD80TM), and to express staphylococcus enterotoxin A (SEA) on the membrane of hepatocellular carcinoma (HCC) cell to form a superantigen gene modified tumor vaccine for HCC . METHODS: SEA and linker-CD80TM gene were amplified through PCR from plasmid containing cDNA of SEA and CD80 . Gene fragments were then subcloned into the multiple cloning sites of retroviral vector pLXSN . Recombinant plasmid was transferred into HepG2 cells mediated with lipofectamine, positive clones were selected in culture medium containing G418 . RT-PCR and indirect immunofluorescence studies confirmed that SEA was expressed specifically on HCC cell membrane . INFgamma-ELISPOT study demonstrated that SEA protein was expressed on the membrane of HCC cells . Cytotoxicity of HepG2-SEA primed CTLs (SEA-T) was analyzed by (51)Cr release assay . T cells cultured with rhIL-2 (IL-2-T) were used as control . RESULTS: Restriction digestion and sequence analyses confirmed the correctness of length, position and orientation of inserted fusion genes . SEA was expressed on the surface of HepG2 cells, HepG2-SEA had strong stimulating effect on production of HepG2 specific CTL (P<0.001) . SEA-T had enhanced cytotoxicity to HepG2 cells (P<0.05) . CONCLUSION: Tumor cell membrane expressed superantigen can be used to reinforce the immune effect of tumor cell vaccine for HCC, which provides a new method of the enhanced active immunotherapy for HCC.

J Comp Pathol, 2004 Jan, 130(1), 70 - 4
Experimentally induced teat stenosis in dairy ewes: clinical, pathological and ultrasonographic features; Mavrogianni VS et al.; A strain of Staphylococcus chromogenes was introduced into the teat cistern of five ewes, teat inflammation and stenosis being the primary consequences . Initially, the inoculated teats were swollen and warm; later, a hard structure was palpated running lengthwise inside the teat, with a thick ring above the tip of the teat, which interfered with expression of milk . Mastitis, confirmed by clinical, cytological, bacteriological and histological findings, was evident 4 days after infection . Ultrasonographically, a hyperechoic line under the mucosa of the teat cistern was observed . At necropsy, the duct wall of the inoculated teats was found to be thickened . Histopathological features included leucocytic infiltration, especially under the mucosa of the teat, and extensive fibrosis in the subcutaneous tissues . S . chromogenes was recovered from scrapings from the duct and the cistern of the inoculated teats.

Int J Hematol, 2003 Nov, 78(4), 335 - 6
Rituximab treatment for relapsed autoimmune hemolytic anemia in Evans syndrome; Galor A et al.; A case is reported of idiopathic relapsed autoimmune hemolytic anemia successfully treated with rituximab . A 43-year-old white male patient with past medical history of Evans syndrome was found to have recurrent autoimmune hemolytic anemia . Previous treatments included steroids, splenectomy, intravenous immunoglobulin, plasmapheresis, staphylococcal Protein A immunoadsorption (Prosorba column), and chemotherapeutic agents (cytoxan and vincristine) . Rituximab was given weekly at 375 mg/m2 for 4 doses . The drug was well tolerated and the patient remains in remission 9 months after completion of therapy.

J Pediatr (Rio J), 1997 May-Jun, 73(3), 161 - 5
{Coagulase-negative Staphylococcus sp . in blood cultures from infants less than 60 days old: infection versus contamination}; Silbert S et al.; OBJECTIVE: The aim of this study was to determine the prevalence of infection versus the prevalence of contamination in infants less than 60 days old who had blood cultures positive for CoNS . MATERIAL AND METHODS: Between February and June - 1993, a number of 45 blood cultures positive for CoNS from 41 patients were studied . Patients were classified in three groups according to their clinical and laboratorial data: I- infected patients, II- non-infected patients and III- dubious.RESULTS: The results showed that 11 patients (26.8%) were included in group I, 25 (61%) in group II (contaminated blood cultures), and 5 (12.2%) in group III . CONCLUSION: It is suggested that more than one blood culture should be requested before treatment with antimicrobials, avoiding unnecessary administration of antibiotics.

OMICS, 2003 Winter, 7(4), 401 - 9
Genetic variations in peripheral blood mononuclear cells in piglets used as an animal model for staphylococcal enterotoxin exposures; Hammamieh R et al.; We have used piglets as an animal model for studying the toxic effects of staphylococcal enterotoxins (SEs) . Piglets are easy to handle, easy to carry out vital measurements, inexpensive, and more importantly, express remarkably similar pathological symptoms and responses to SE intoxication as humans at comparable doses . Microarray analyses are used to study the effect of many infections on gene expression profile in peripheral blood mononuclear cells . This high throughput application offers detailed depiction of alteration at the molecular levels . When using high throughput gene expression analysis, there is a high possibility of finding genes that vary normally in the tissues under study . It is necessary to verify genes that are normally differentially expressed between piglets . To evaluate the normal physiological variation in gene expression in vivo in piglets, we used cDNA microarray to measure gene expression levels in peripheral blood mononuclear cells from 10 normal Yorkshire piglets . We used analysis of variance to determine genes that showed statistically significant variations across piglets . Out of 1185 genes, 19 (1.6%) genes revealed statistically significant variance between RNA samples . Some of these varying genes are involved in stress response, immune response, and transcription . This study facilitates the characterization of gene expression base line needed for meaningful interpretation of microarray data.

Shock, 2004 Jan, 21(1), 77 - 85
In vivo and in vitro cytokine modulatory activity of newly synthesised 2-aminotetraline derivatives; Ruggiero V et al.; In the present study, the protective effect of newly synthesised 2-aminotetralines was investigated in murine models of toxic shock . A few derivatives protected mice against lethality induced by lipopolysaccharide from different bacterial strains and shock induced by staphylococcal enterotoxin B in mice sensitized by D-Galactosamine (D-Galn) . Notably, one derivative, S(-)-2-amino-6-fluoro-7-methoxy-1,2,3,4 tetrahydronaphthalene hydrochloride (ST1214), was also effective when administered orally (30 mg kg-1) in a therapeutic regimen . ST1214 markedly inhibited the production of the proinflammatory cytokines, such as tumor necrosis factor-alpha (TNF-alpha), Interleukin-1beta (IL-1beta), Interleukin-12 (IL-12), interferon-gamma (IFN-gamma), as well as the inflammatory mediator nitric oxide (NO), and concurrently enhanced the production of the anti-inflammatory cytokine IL-10 . Moreover, ST1214 dose-dependently reduced TNF-alpha production by human peripheral blood mononuclear cells and promonocytic THP-1 cells in vitro . In the latter, ST1214 was found to inhibit lipopolysaccharide-induced TNF-alpha secretion but not cytokine mRNA accumulation . These results suggest that the mechanism of action of ST1214 involves blockade of posttranscriptional events of TNF-alpha production, apparently independent of p38 and ERK kinase activity . These results show beneficial effects of 2-aminotetralines in murine shock models and indicate a distinct counter-regulatory activity in down-regulating proinflammatory cytokine response, and upregulating IL-10 . One derivative, i.e., ST1214, can be regarded as a lead compound in the development of novel drugs effective in anti-inflammatory strategies.

Proc Natl Acad Sci U S A, 2003 Dec 23, 100(26), 15498 - 503 Epub 2003 Dec 15.
Electroosmotic enhancement of the binding of a neutral molecule to a transmembrane pore; Gu LQ et al.; The flux of solvent water coupled to the transit of ions through protein pores is considerable . The effect of this electroosmotic solvent flow on the binding of a neutral molecule {beta-cyclodextrin (betaCD)} to sites within the staphylococcal alpha-hemolysin pore was investigated . Mutant alpha-hemolysin pores were used to which betaCD can bind from either entrance and through which the direction of water flow can be controlled by choosing the charge selectivity of the pore and the polarity of the applied potential . The Kd values for betaCD for individual mutant pores varied by >100-fold with the applied potential over a range of -120 to +120 mV . In all cases, the signs of the changes in binding free energy and the influence of potential on the association and dissociation rate constants for betaCD were consistent with an electroosmotic effect.

J Pharmacol Toxicol Methods, 2004 Jan-Feb, 49(1), 39 - 55
The automated bioaerosol exposure system: preclinical platform development and a respiratory dosimetry application with nonhuman primates; Hartings JM et al.; INTRODUCTION: Well-characterized inhalation exposure systems are critical for preclinical testing and pathogenesis studies . The automated bioaerosol exposure system (ABES) provides a microprocessor-driven inhalation platform that provides exquisite data acquisition and control over all aspects of inhalation exposures . Because this represents a new technology, the development and characteristics of the ABES are thoroughly discussed . In addition to control over homeostatic and aerosol conditions, the ABES incorporates a dosimetry function based on respiratory performance of the test animal during inhalation . METHODS: To test the system, rhesus macaques were initially sham-exposed using the ABES in a head-only inhalation configuration . The ABES was subsequently used under biosafety level (BSL)-III conditions in a vaccine efficacy challenge using aerosolized staphylococcal enterotoxin B (SEB) toxin, again using the real-time dosimetry function of the system . RESULTS: Sham exposure results indicated significant departures from corresponding whole-body plethysmography (WBP) respiratory function estimates taken before the inhalation procedure . The results of the SEB exposure demonstrated the utility of using the ABES to generate consistently accurate and precise inhalation dose . DISCUSSION: Taken together, the results of the sham and toxin challenge experiments demonstrate that the dosimetry function of the ABES improves the precision and accuracy of inhaled dose delivery and calculation as compared to predictive WBP conducted before the exposure . The ABES represents a highly adaptable platform for the design of inhalation systems to suit the requirements of a variety of animal models.

Analyst, 2003 Oct, 128(10), 1275 - 80 Epub 2003 Sep 12.
Rapid polyelectrolyte-based immunofiltration technique for testosterone detection in serum samples; Zherdev AV et al.; A new immunofiltration assay for testosterone is proposed . During the first step of the assay, testosterone molecules in serum samples compete in solution with the testosterone-peroxidase conjugate for interaction with anti-testosterone antibodies pre-bound to the conjugate between staphylococcal protein A and polymethacrylate polyanion . The reaction mixture is then filtered through a membrane charged with immobilized poly(N-ethyl-4-vinylpyridinium) polycation . The filtration is accompanied by a rapid separation of the polyanion containing complexes due to high-affinity electrostatic interactions . Following removal of unbound compounds the immobilized peroxidase is detected using a substrate that produces an insoluble coloured product . The proposed assay has been shown to combine high speed (20 min) and sensitivity (0.1 ng ml(-1)), and to be applicable for out-of-laboratory conditions . Based on densitometric measurements, the RSD of the assay is calculated to be 3.2-5.1% (n = 4) . The proposed assay is 4 times faster than the microplate enzyme immunoassay (ELISA) based on the same immunoreagents . Pre-incubation of the antibody and the polyanion-protein A conjugate at a certain ratio excludes the influence of immunoglobulins from the tested serum samples on the assay results . The polyanion-protein A conjugate can be used as a universal reagent, eliminating the necessity to modify specific antibodies for each immunoassay.

Syst Appl Microbiol, 2003 Nov, 26(4), 601 - 10
Design and evaluation of specific PCR primers for rapid and reliable identification of Staphylococcus xylosus strains isolated from dry fermented sausages; Blaiotta G et al.; Rapid and reliable identification of Staphylococcus xylosus was achieved by species-specific PCR assays . Two sets of primers, targeting on xylulokinase (xylB) and 60 kDa heat-shock protein (hsp60) genes of S . xylosus, respectively, were designed . Species-specificity of both sets of primers was evaluated by using 27 reference strains of the DSM collection, representing 23 different species of the Staphylococcus genus and 3 species of the Kocuria genus . Moreover, 90 wild strains isolated from different fermented dry sausages were included in the analysis . By using primers xylB-F and xylB-R the expected PCR fragment was obtained only when DNA from S . xylosus was used . By contrast, amplification performed by using primers xylHs-F and xylHs-R produced a single PCR fragment, of the expected length, when DNA from S . xylosus, S . haemolyticus, S . intermedius and S . kloosii were used as template . Nevertheless, AluI digestion of the xylHs-F/xylHs-R PCR fragment allowed a clear differentiation of these 4 species . The rapidity (about 4 h from DNA isolation to results) and reliability of the PCR procedures established suggests that the method may be profitably applied for specific detection and identification of S . xylosus strains.

Arch Inst Pasteur Tunis, 1999 Jan-Apr, 76(1-4), 27 - 31
{Preparation and testing of Sardinella peptones: application to lipase production by Staphylococcus sp}; Ellouz Y et al.; Production of lipase by Staphylococcus sp . in media containing fish peptones from sardinelle (Sardinella aurita) prepared in the laboratory was studied . Lipase production is strongly affected by lipids present in fish flours . Fish peptones prepared from dIgresed whole flesh was an excellent substrate for lipase production . A comparison of lipase production in media containing fish peptones or high quality commercial peptones indicated that fish peptones enhanced enzyme formation.

Orv Hetil, 2003 Oct 26, 144(43), 2115 - 20
{Meropenem therapy in children with febrile neutropenia}; Muller J et al.; OBJECTIVES: Infectious complications are the major causes of morbidity and mortality in children receiving chemotherapy for malignant diseases . Granulocytopenia carries the risk of bacterial infection, and also, if prolonged, of fungal infection . The aim of this study was to evaluate the clinical effectiveness of meropenem in immunocompromised children in association with isolated bacteria from blood cultures and clinical background . METHODS: Retrospective study of all febrile episodes when meropenem was used in neutropenic children between January 1998 and December 2002 in the haemato-oncological units of the authors hospital . During the study period meropenem was used in 87 febrile events diagnosed in 55 patients (mean age 10 years 5 months), and 328 bacterial cultures were performed . Blood samples were taken from each patient with granulocytopenia (< 0.5 G/l) and fever (> or = 38 degrees C), prior to the start of any antibiotic therapy . For the microbiological process Bactec 9050 (Becton Dickinson) blood culture systems were used . RESULTS: Microorganisms were detected and identified in 64 (19.5%) from the 328 hemocultures . There was a predominance of Gram-positive strains, 67% (43/64)--the most common bacteria being coagulase negative Staphylococcus (cnS) . From the 87 periods in 43 cases (49.4%) the infection was documented microbiologically . In 16 additional cases the infection was proven clinically (based on the clinical course, laboratory and radiologic results) and 32.2% (28/87) of the febrile neutropenic episodes were considered to be fever of unknown origin . Meropenem was used in a mean dose of 60.8 (30-120) mg/kg/die, for 9.3 (2-24) days . The success rate of the meropenem therapy -excluding the proven fungal (n = 13) or cnS (n = 15) infections-was 72.9% . No severe side effects occurred in any regimens . CONCLUSION: The results demonstrate that meropenem is effective and well-tolerated when used for the treatment of feverish neutropenic cancer children.

Proc Natl Acad Sci U S A, 2003 Dec 9, 100(25), 15035 - 40 Epub 2003 Dec 01.
Control of Bcl-2 expression by reactive oxygen species; Hildeman DA et al.; Reactive oxygen species (ROS) mediate apoptosis in many different cell types . We have previously shown that the antioxidant Mn(III) tetrakis(5,10,15,20-benzoic acid)porphyrin (MnTBAP) decreased intracellular ROS and prevented the apoptosis of activated T cells in vitro . To determine the mechanism(s) by which MnTBAP afforded such protection, we used Affymetrix (Santa Clara, CA) gene arrays to compare gene expression in T cells activated with staphylococcal enterotoxin B in vivo then cultured with or without MnTBAP . This analysis showed that the antioxidant increased the expression of Bcl-2, an antiapoptotic molecule whose levels are normally decreased by T cell activation . Culture with MnTBAP revealed a tight inverse correlation between the levels of Bcl-2 and ROS within T cells . In vivo, production of ROS in activated T cells occurred before Bcl-2 down-regulation . Furthermore, MnTBAP's ability to prevent death required the expression of Bcl-2 in most T cells . Finally, neither ROS production nor the effects on Bcl-2 expression required Bim, the Bcl-2 antagonist that mediates the death of activated T cells in vivo . Taken together, our results suggest that ROS sensitize T cells to apoptosis by decreasing expression of Bcl-2.

Brain Behav Immun, 2004 Jan, 18(1), 46 - 54
Effects of bacterial superantigens on behavior of mice in the elevated plus maze and light-dark box; Rossi-George A et al.; Bacterial superantigens, such as staphylococcal enteroxins A and B (SEA/SEB) stimulate T cells to produce high levels of cytokines in blood . Previously it had been shown that these toxins were capable of stimulating increased neuroendocrine activity and enhanced behavioral reactivity to novel gustatory and non-gustatory stimuli . Therefore, it was suggested that these superantigens may promote anxiety-like behavior . In the current set of experiments, BALB/cByJ and C57BL/6J male mice were challenged with either SEB (50 microg) or SEA (5 or 10 microg) and tested for behavior in the elevated plus maze (EPM) . Results suggested an absence of increased anxiety-like behavior, with exploration of the open arms being enhanced by SEA or SEB treatment . In another test of anxiety, the light-dark box, SEB challenge of BALB/cByJ mice 90 min prior to testing, did not alter exit latency, activity nor time spent in the dark . However, in a second experiment, it was found that if animals were first tested for consumption, followed by testing in the light-dark box, SEB challenged animals displayed increased exit latency and reduced exploration . These studies suggest that in standard tests of rodent anxiety-like behavior, evidence for the induction of anxiety-like processes subsequent to challenge with SEA or SEB is not patently discernable . However, neurobiological events induced by immunological challenge might synergize with reactivity to psychogenic and/or gustatory stimuli, thereby resulting in increased anxiety-like behavior that could be unmasked by standard behavioral tests such as the light-dark box or EPM.

J Am Chem Soc, 2003 Dec 3, 125(48), 14841 - 6
Ab initio folding of helix bundle proteins using molecular dynamics simulations; Jang S et al.; We have demonstrated that ab initio fast folding simulations at 400 K using a GB implicit solvent model with an all-atom based force field can describe the spontaneous formation of nativelike structures for the 36-residue villin headpiece and the 46-residue fragment B of Staphylococcal protein A . An implicit solvent model combined with high-temperature MD makes it possible to perform direct folding simulations of small- to medium-sized proteins by reducing the computational requirements tremendously . In the early stage of folding of the villin headpiece and protein A, initial hydrophobic collapse and rapid formation of helices were found to play important roles . For protein A, the third helix forms first in the early stage of folding and exhibits higher stability . The free energy profiles calculated from the folding simulations suggested that both of the helix-bundle proteins show a two-state thermodynamic behavior and protein A exhibits rather broad native basins.

Support Care Cancer, 2004 Feb, 12(2), 99 - 105 Epub 2003 Nov 25.
Totally implantable subcutaneous port system versus central venous catheter placed before induction chemotherapy in patients with acute leukaemia-a randomized study; Johansson E et al.; A group of 43 adult patients with acute leukaemia (AL) were randomized to receive a double-lumen totally implantable subcutaneous port system (PORT, n=19) or a double-lumen central venous catheter (CVC, n=24) before induction chemotherapy . Six patients were excluded due to protocol violation ( n=4, CVC) and technical difficulties ( n=2, PORT) . A standardized catheter record form was used for recording of catheter function, local infection and bleeding . The study was prematurely closed due to extensive subcutaneous bleeding after placement in five patients with a PORT . Intention to treat ( n=43) or per protocol (PP) analysis ( n=37) did not reveal a significant difference between the two groups with regard to catheter survival time (PP PORT, median 113 days, range 2-634 days; CVC, 55 days, 11-223 days) . The number of positive blood cultures per 100 central venous access device days was significantly higher in the CVC group (median 3.6 per 100 days) than in the PORT group (0.9 per 100 days; P=0.02) . In addition, the time to the first blood culture positive for coagulase-negative staphylococcus was shorter in the CVC group (median 14 days) than in the PORT group (52 days; P=0.02) . Despite fewer infectious complications in the PORT group the use of a double-lumen CVC is advocated in patients with AL undergoing induction treatment due to the risk of extensive local bleeding after placement of the PORT.

J Gen Virol, 2003 Dec, 84(Pt 12), 3453 - 7
Cytokine profiles of peripheral blood mononuclear cells from pigs with postweaning multisystemic wasting syndrome in response to mitogen, superantigen or recall viral antigens; Darwich L et al.; In vitro cytokine profiles of peripheral blood mononuclear cells (PBMC) from pigs with postweaning multisystemic wasting syndrome (PMWS) and healthy pigs were determined in response to recall viral antigens (porcine circovirus type 2; PCV2), mitogens (phytohaemagglutinin) or superantigens (staphylococcal enterotoxin B) . PBMC from PMWS-affected pigs, in contrast to those from healthy pigs, responded to recall PCV2 antigen by releasing IL-10 and IFN-gamma, but they were less able or even unable to produce IL-4, IL-2 or IFN-gamma upon challenge with mitogen or superantigen . Moreover, only PCV2 had the ability to downregulate or suppress the release of IL-4 and IL-2 from PBMC from both healthy and diseased animals, and to stimulate the production of pro-inflammatory cytokines (IL-1beta, IL-8) . In conclusion, the immune system cells of PMWS pigs have a diminished ability to perform their immunological functions upon viral or immunostimulatory molecules . In addition, PCV2 can alter the functionality of PBMC in both healthy and PMWS pigs.

J Nat Prod, 2003 Nov, 66(11), 1495 - 8
New bromotyrosine derivatives from an association of two sponges, Jaspis wondoensis and Poecillastra wondoensis; Park Y et al.; Three new bromotyrosine derivatives (4-6) were isolated from an association of two sponges, Jaspis wondoensis and Poecillastra wondoensis, along with the previously described (E,E)-psammaplin A (1), (E,Z)-psammaplin A (2), psammaplin D (3), bisaprasin (7), and (3-bromo-4-hydroxyphenyl)acetonitrile (8) . The structures of the new compounds were established on the basis of NMR and MS spectroscopic analysis . The compounds 1, 3, and 5-7 displayed significant cytotoxicity against human lung (A549), ovarian (SK-OV-3), skin (SK-MEL-2), CNS (XF498), and colon (HCT15) cancer cell lines . Compounds 3-7 were further evaluated for antibacterial activity against methicillin- or ofloxacin-resistant Staphylococcus strains . Compound 4 exhibited more potent antibacterial activity than meropenem against several strains.

Clin Nephrol, 2003 Nov, 60(5), 327 - 34
Interleukin-18, interleukin-18 binding protein and impaired production of interferon-gamma in chronic renal failure; Lonnemann G et al.; Uremia is associated with suppressed cellular immune responses, manifested, in part, by impaired interferon-gamma (IFNgamma) production . We investigated the influence of kidney function on plasma levels of interleukin-18 binding protein (IL-18BP), the naturally occurring inhibitor of IL-18 . METHODS: Plasma levels of IL-18, IL-18BP and IFNgamma were measured by specific immunoassays in patients with normal kidney function (NKF, n = 29), in patients with chronic renal insufficiency (CRI, n = 29), and in patients on hemodialysis (HD, n = 40) . In addition, Staphylococcus epidermidis-induced production of IFNgamma and IL-18 in whole blood cultures was determined in 12 patients on HD and compared to production in 9 controls with NKF . RESULTS: Plasma IL-18 (mean +/- SEM) in NKF was 17.9 +/- 3.6 pg/ml, in CR142.6 +/- 7.0 pg/ml (p < 0.01), and in HD 93.5 +/- 13.6 pg/ml (p < 0.001) . The level of IL-18BP in NKF was 3.4 +/- 0.4 ng/ml, in CRI 7.5 +/- 0.7 ng/ml (p < 0.001), and in HD 13.1 +/- 0.8 ng/ml (p < 0.001) . Plasma IL-18BP was inversely correlated with creatinine clearance (correlation coefficient: -0.7479) . The level of free IL-18 was calculated in NKF to be 13.8 +/- 3.3 pg/ml, in CRI 23.6 +/- 3.9 pg/ml (not significant), and in HD 39.6 +/- 5.9 pg/ml (p < 0.001) . Stimulated whole blood production of IFNgamma in NKF was 185 +/- 74 pg/10(6) mononuclear cells (PBMC), but suppressed in HD to 27.3 +/- 16 pg/10(6) PBMC (p < 0.05) . CONCLUSION: In uremia, retention of IL-18BP does not suffice to neutralize most of the concomitantly raised levels of total IL-18 resulting in elevated levels of free IL-18 . Nevertheless, IFNgamma production in whole blood is reduced in patients on HD . Therefore, suppression of IFNgamma production in uremia may be due to inhibitors of IFNgamma production other than IL-18BP.

Proc Natl Acad Sci U S A, 2003 Dec 9, 100(25), 14812 - 6 Epub 2003 Nov 24.
Atomically detailed folding simulation of the B domain of staphylococcal protein A from random structures; Vila JA et al.; The conformational space of the 10-55 fragment of the B-domain of staphylococcal protein A has been investigated by using the electrostatically driven Monte Carlo (EDMC) method . The ECEPP/3 (empirical conformational energy program for peptides) force-field plus two different continuum solvation models, namely SRFOPT (Solvent Radii Fixed with atomic solvation parameters OPTimized) and OONS (Ooi, Oobatake, Nemethy, and Scheraga solvation model), were used to describe the conformational energy of the chain . After an exhaustive search, starting from two different random conformations, three of four runs led to native-like conformations . Boltzmann-averaged root-mean-square deviations (RMSD) for all of the backbone heavy atoms with respect to the native structure of 3.35 A and 4.54 A were obtained with SRFOPT and OONS, respectively . These results show that the protein-folding problem can be solved at the atomic detail level by an ab initio procedure, starting from random conformations, with no knowledge except the amino acid sequence . To our knowledge, the results reported here correspond to the largest protein ever folded from a random conformation by an initial-value formulation with a full atomic potential, without resort to knowledge-based information.

Jpn J Ophthalmol, 2003 Nov-Dec, 47(6), 609 - 11
Evaluation of staphylococcal enterotoxin-specific IgE antibody in tears in allergic keratoconjunctival disorders; Shoji J et al.; PURPOSE: To investigate the presence of staphylococcal enterotoxin A (SEA) and B (SEB)-specific IgE antibodies in tears from patients with allergic conjunctival disorders . METHODS: The study included 8 eyes of 4 patients with perennial allergic conjunctivitis (PAC), 14 eyes of 7 patients with vernal keratoconjunctivitis (VKC), 12 eyes of 6 patients with atopic keratoconjunctivitis (AKC), and 10 eyes of 10 healthy volunteers as controls . Tears were sampled by the method of the Schirmer test I . Sampled tears were eluted and SEA- and SEB-specific IgE antibodies were analyzed by the AlaSTAT-IMMULIZE method . RESULTS: SEA-specific IgE antibodies in tears were positive in 9 of 14 eyes in VKC patients and in 1 of 12 eyes in AKC patients . SEB-specific IgE antibodies in tears were positive in 7 of 14 eyes in VKC patients and in 2 of 12 eyes in AKC patients . Values for antibodies were higher in patients with severe clinical findings . However, all the cases in the normal control and the PAC groups were negative for both antibodies . CONCLUSION: Our data strongly suggested that staphylococcal enterotoxin may cause type I allergy, and may be an exacerbating factor for vernal keratoconjunctivitis and atopic keratoconjunctivitis.

Anal Chem, 2003 Aug 15, 75(16), 4043 - 9
Preparation of quantum dot-biotin conjugates and their use in immunochromatography assays; Lingerfelt BM et al.; Biotinylated, highly luminescent CdSe-ZnS quantum dot (QD) conjugates were prepared and used in immunofiltration assays . Water-soluble quantum dot surfaces having a homogeneous negative charge density at basic pH were initially coated with a two-domain recombinant maltose-binding protein appended with a positively charged leucine zipper . Biotin functionalization of these electrostatically stabilized QD-protein complexes was then carried out using amine-reactive NHS biotin . These protein-coated biotin-functionalized quantum dot conjugates were incorporated into flow immunofiltration/displacement assays employing Affi-gel agarose resin for antibody immobilization, analyte capture, and immune complex formation with a second biotinylated antibody . A key component of the assay was the use of tetranitromethane-modified NeutrAvidin, used to link the biotinylated QDs to the immune complexes and facilitate their release at basic pH for subsequent quantification . This assay methodology was used to detect as little as 10 ng/mL staphylococcal enterotoxin type-B.

Protein Eng, 2003 Nov, 16(11), 831 - 40
Effect of mutations involving charged residues on the stability of staphylococcal nuclease: a continuum electrostatics study; Borjesson U et al.; A continuum electrostatics model is used to calculate the relative stabilities of 117 mutants of staphylococcal nuclease (SNase) involving the mutation of a charged residue to an uncharged residue . The calculations are based on the crystallographic structure of the wild-type protein and attempt to take implicitly into account the effect of the mutations in the denatured state by assuming a linear relationship between the free energy changes caused by the mutation in the native and denatured states . A good correlation (linear correlation coefficient of approximately 0.8) is found with published experimental relative stabilities of these mutants . The results suggest that in the case of SNase (i) charged residues contribute to the stability of the native state mainly through electrostatic interactions, and (ii) native-like electrostatic interactions may persist in the denatured state . The continuum electrostatics method is only moderately sensitive to model parameters and leads to quasi-predictive results for the relative mutant stabilities (error of 2-3 kJ mol(-1) or of the order of k(B)T), except for mutants in which a charged residue is mutated to glycine.

Arch Biochem Biophys, 2003 Dec 1, 420(1), 87 - 94
Effect of a specific hydrogen bond (N138ND2-Q106O) on conformational integrity, stability, and activity of staphylococcal nuclease; Huang S et al.; There are two hydrogen bonding interactions (N138ND2-Q106O and Y54OH-S141OG) between the C-terminal region and the main body of staphylococcal nuclease (SNase) . To examine the role of these hydrogen bonds, SNase(141) and its three mutants, SNase(141)N138D, SNase(141)S141A, and SNase(141)N138D/S141A, were created . The N138D mutation has the N138ND2-Q106O interaction deleted and the S141A mutation has the Y54OH-S141OG and S141OG-N138O interactions deleted . The conformational features, stability, and activity of the proteins have been compared by using circular dichroism, intrinsic and ANS-binding fluorescence, GdnHCl-induced denaturation, and activity assay . The results clearly show that the N138D mutation significantly alters the secondary and tertiary structures of the protein, producing a partially unfolding state; in contrast, the S141A mutation has no such effect on structure . These results strongly suggest that the specific hydrogen bond, N138ND2-Q106O, plays an important role in maintaining the conformational integrity and stability of the nuclease.

Transpl Int, 1992, 5 Suppl 1, S589 - 93
Factors involved in peripheral T cell tolerance: the extent of clonal deletion or clonal anergy depends on the age of the tolerized lymphocytes; Kuschnaroff LM et al.; After injection of SEB (staphylococcus enterotoxin B), normal adult mice, or thymectomized irradiated mice (TX irr.) reconstituted with lymphocytes taken from normal adult mice became specifically tolerant of SEB . At the same time the percentage of Vbeta8 positive CD4 lymphocytes known to be responsive to SEB was almost 50% decreased, indicating that a high level of clonal deletion was realized . In contrast, mice with an exclusively old T cell compartment (old thymectomized mice, TX irr . mice reconstituted several months previously) became tolerant of SEB without deleting their Vbeta8 + CD4 + cells, indicating that clonal anergy was the major mechanism in play in the induction of tolerance . Finally, TX irr . mice reconstituted with single positive thymocytes known to become recent thymic emigrants developed tolerance for SEB together with a high level (70%) of clonal deletion . Altogether these results indicated that the mechanism involved in peripheral tolerance depended on the age of the lymphocyte: very young lymphocytes underwent mainly clonal deletion whereas long lived lymphocytes underwent predominantly clonal anergy.

Acta Orthop Scand, 2003 Oct, 74(5), 617 - 21
Occurrence of ica genes for slime synthesis in a collection of Staphylococcus epidermidis strains from orthopedic prosthesis infections; Arciola CR et al.; Staphylococcus epidermidis is a frequent pathogen in infections associated with orthopedic implants . We studied 123 S . epidermidis strains from infections related to orthopedic implants, as regards their ability to express a factor of virulence, namely the slime, an extracellular polysaccharide, which mediates adherence to implants and bacterial colonization . The slime-producing ability was determined by PCR detection of icaA and icaD genes responsible for slime synthesis, and by culture on Congo red agar plates in which slime-producing strains form black colonies, while nonslime-forming ones develop red colonies . 56% of the S . epidermidis isolates were icaA- icaD-positive and grew to become black colonies . In the evaluation of the distribution of slime-forming strains in different sites and types of implants, we found a slight, but not statistically significant, increase in slime-forming strains in total joint prostheses, where tissue compression near the articular faces can form niches in which bacteria crowd, sheltered by the slime . Our findings confirm the role of ica genes as a virulence marker in the pathogenesis of implant-associated orthopedic infections . However, they do not show the existence of a higher frequency of slime-positive strains in a specific type of implant.

Indian J Pediatr, 2003 Sep, 70(9), 743 - 5
Right-sided diaphragmatic hernia masquerading as staphylococcal pneumonia; Kaur G et al.; A 9-month-old infant who was diagnosed to have right-sided diaphragmatic hernia with no other associated anomalies, is being reported here . He had presented with cough for one month and respiratory difficulty for one-week duration and history of bilious vomiting two days prior to admission . His chest X-ray showed multiple lucent shadows in the right lower zone in A-P and lateral views, with not very well defined diaphragm on the same side . A diagnosis of staphylococcal pneumonia was considered but with clinical and radiological picture, a right-sided diaphragmatic hernia was strongly suspected which was confirmed by doing fluoroscopy and CT-scan of the chest . He underwent a successful correction of the defect . So radiological finding of pleural effusion with displacement of mediastinum to the left and the presence of bowel gas high in the right upper quadrant should alert the possibility of a right-sided Bochdalek hernia.

Ophthalmologe, 2003 Oct, 100(10), 825 - 31 Epub 2003 Jun 19.
{The influence of a new surface treatment of silicone intracoular lenses with fluoralkylsitan on the adherence of endophthalmitic bacteria in vitro}}; Kienast A et al.; INTRODUCTION: Dynasilan is a fluoroalkylsilan which is able to bind to surface active molecules of intraocular lenses (IOLs), thereby offering a new option for surface modification of silicone lenses . The purpose of this in vitro study was to investigate the influence of this new surface treatment on the adherence of two typical endophthalmitis-inducing bacteria ( Staphylococcus epidermidis, Propionibacterium acnes) . MATERIALS AND METHODS: A total of 14 Dynasilan-treated and 14 untreated silicone lenses were incubated at 37 degrees C for 24 h in brain heart infusion broth (10(8) CFU/ml) either with Staphylococcus epidermidis or with Propionibacterium acnes for 1 h . Subsequently, the adherent bacteria were resuspended using ultrasonification at 35 kHz for 3 x 45 s . After a dilution series and incubation at 37 degrees C for 24 h or 3 days the colonies were counted . RESULTS: On untreated IOLs incubated with Staphylococcus epidermidis the average number of bacteria was 3.6 x 10(7)/ml, and on treated IOLs the number of counted colonies was reduced to 1.09 x 10(7)/ml . Incubated with Propionibacterium acnes the average number of adherent bacteria on untreated IOLs was 4.75 x 10(4)/ml and on modified IOLs the number was reduced to 2.94 x 10(4)/ml . CONCLUSION: Dynasilan surface treatment may reduce the adherence of Staphylococcus epidermidis and Propionibacterium acnes on silicone intraocular lenses . Further studies regarding the stability of this treatment, its biocompatibility and influence on lens epithelial cell adhesion are in progress.

Transfusion, 2003 Nov, 43(11), 1508 - 13
Immunoadsorption may provide a cost-effective approach to management of patients with inhibitors to FVIII; Freedman J et al.; BACKGROUND: Immunoadsorption of plasma with Staphylococcal protein A removes immunoglobulins and immune complexes; hence, it should effectively remove inhibitors to FVIII in acquired or congenital hemophilia . The procedure may be cost effective, given the expense of therapies used to treat patients with inhibitors, particularly in an acute setting . STUDY DESIGN AND METHODS: Three patients with inhibitors to FVIII were treated with the Excorim Immunosorba system (two columns used in tandem) . Costs for immunoadsorption and for other therapeutic products administered to the patients were calculated . RESULTS: Two patients had acquired hemophilia and severe bleeding associated with low levels of circulating FVIII and high levels of inhibitors to FVIII . They failed to achieve a satisfactory response to management with immunosuppression, pFVIII, recombinant FVIIa or IVIG but responded rapidly, with long-term benefit, to immunoadsorption therapy . The third patient had congenital hemophilia and immunoadsorption was effective in reducing his inhibitor level, allowing him to undergo immune tolerance therapy . Costs of treatment before immunoadsorption were markedly higher than those associated with the immunoadsorption procedures (i.e., >Can 350,000 dollars and >Can 1,000,000 dollars vs . < 20,000 dollars) . CONCLUSION: Immunoadsorption appears to be an effective and cost-effective alternative in the management of patients with inhibitors to FVIII.

FEMS Microbiol Lett, 2003 Nov 7, 228(1), 143 - 9
Preliminary characterization of beta-decarboxylase activities in Staphylococcus carnosus 833, a strain used in sausage fermentation; Fadda S et al.; In sausage, Staphylococcus carnosus increases the levels of methyl ketones which could arise from incomplete beta-oxidation of fatty acids followed by a decarboxylation . The objective of this work was to characterize the beta-decarboxylase activities in cell-free extract . By using different substrates, at least two kinds of beta-decarboxylase activities were shown: an acetoacetate-like and an oxaloacetate-like . The first one leads to the production of ketones from ethylbutyryl acetate or acetoacetic acid . The activity was optimal at pH 6.0, stimulated by pyridoxal phosphate but ethylenediamine tetraacetic acid (EDTA), NaCl, iodoacetate and curing additives were inhibitory . The second decarboxylase activity leads to the production of pyruvic acid from oxaloacetic acid . This activity was optimal at pH 5.0 and stimulated by divalent ions and biotin.

Nucleosides Nucleotides Nucleic Acids, 2003 Oct, 22(10), 1859 - 66
Study on the interaction between nucleic acids and acetamiprid; Jie N et al.; The interaction between deoxyribonucleic acid (DNA) and acetamiprid was studied . It was found that the fluorescence of acetamiprid could be enhanced in the presence of DNA in sulfuric acid solution . The excitation and emission wavelength of acetamiprid was 291 nm and 587 nm, respectively . Under optimal conditions, the calibration graph is over the range of 0.1-10 micromL(-1) . The calibration limit is 0.06 microg mL(-1) (S/N = 3) . The determination results of DNA in yeast cell and golden staphylococcus samples by this method were satisfactory . The mechanism of the reaction is discussed.

Clin Diagn Lab Immunol, 2003 Nov, 10(6), 1011 - 8
Inflammatory responses of bovine polymorphonuclear neutrophils induced by staphylococcal enterotoxin C via stimulation of mononuclear cells; Kuroishi T et al.; To elucidate the pathological roles of staphylococcal enterotoxin C (SEC) in bovine staphylococcal mastitis, a histopathological analysis of SEC-inoculated mammary glands was performed . SEC-inoculated mammary glands exhibited interstitial inflammation, and the leukocytes that migrated into the gland were predominantly polymorphonuclear neutrophils (PMN) . In the gland cistern tissues dissected from SEC-inoculated mammary glands, epithelial cellular degeneration was observed . We also investigated the physiological effects of SEC on PMN in vitro . PMN migration was induced by culture supernatant of SEC-stimulated peripheral blood mononuclear cells (S-PBMC sup) but not by that of nonstimulated PBMC (N-PBMC sup) . The concentration of interleukin-8 was significantly (P < 0.05) higher in S-PBMC sup than N-PBMC sup, and a significantly (P < 0.05) higher mRNA expression of growth-regulated oncogenes was detected in SEC-stimulated PBMC than in nonstimulated PBMC . Milk PMN collected from SEC-inoculated mammary glands produced more than 2 times the amount of superoxide at 1 day postinoculation (dpi) than at 0 dpi in the presence of phorbol 12-myristate 13-acetate (PMA) . PMN cultured with S-PBMC sup for 24 h also produced significantly (P < 0.05) larger amounts of superoxide than those cultured with N-PBMC sup in the presence of PMA . Moreover, S-PBMC sup induced the long-time survival of PMN . These results indicate that SEC induces the activation of PMN via the stimulation of mononuclear cells.

Chest, 2003 Nov, 124(5), 1968 - 74
Evidence for the transmissibility of atopy: hypothesis; Hussain I et al.; The etiology of atopy is unknown . Its family distribution suggests transmissibility . Populations moving from countries with a low incidence to those with a high incidence increase to the higher rate . African and New Guinea village groups developed asthma with return of individuals who have acquired atopy in the city . Protection (and possibly immunity) develops with early exposure to child care or to affected older siblings . T helper (Th) type 2 clones driving specific allergies remain active even without further allergen exposure . Other IgE responses remain normal . Once boosted to completeness, the patterns of skin test results remain quite stable, possibly by the localization of abnormality maintained by immunity . An example of a virus causing the immortality of Th2 cells is herpes simplex virus type 1 . It infects mouse or human Th2 cells and, although it does not multiply, causes immortality by increasing FAS-mediated apoptosis of T cells directed against the infected cells . Human T-cell leukemia virus 1 and probably others use similar ploys . Abnormal levels of FAS receptors and resistance to FAS apoptosis in nasal polyp lymphocytes and abnormal Th2 clones of atopy are interesting in this regard . The localizing role of a staphylococcal superantigen in atopic dermatitis, and possibly in autoimmunity in nonatopic eczema and intrinsic asthma, encourage the consideration of roles for microorganisms in localization and etiology . The epidemiology and characteristics of atopic disease support the plausibility of a viral hypothesis.

Ann Thorac Surg, 2003 Nov, 76(5), 1655 - 60
Mediastinitis after pediatric cardiac surgery: a 15-year experience at a single institution; Tortoriello TA et al.; BACKGROUND: The spectrum of sternal wound infections after cardiac surgery ranges from superficial infections to a deep sternal infection known as mediastinitis . Mediastinitis is a rare but clinically relevant source of postoperative morbidity and mortality in adult and pediatric patients after cardiac surgery . METHODS: We retrospectively identified all patients diagnosed with mediastinitis after cardiac surgery from January 1987 to December 2002 (17 patients/7,616 surgeries = 0.2%) . Demographic data, cardiac diagnosis, cardiac surgery, hospital length of stay, associated medical diagnosis, and surgical treatment for mediastinitis were collected . RESULTS: Fifteen pediatric patients (age < 18 years) were diagnosed with mediastinitis (mean age at diagnosis 37.5 months, range 21 days to 17 years . The median postoperative day of diagnosis was 14 days (6 to 50 days) . The most common organism was Staphylococcus species (n = 9) . Six patients had an associated bacteremia . The median hospital length of stay for all patients was 42.5 days (range 16 to 163 days) . The hospital mortality was 1 of 15 (6%) . Each patient was treated with intravenous antibiotics; sternal debridement; and rectus abdominus flap reconstruction (n = 7), pectoralis muscle flap reconstruction (n = 3), omentum reconstruction (n = 1), or primary sternal closure (n = 4) . Three patients have undergone redo-sternotomy with orthotopic heart transplantation, bidirectional cavopulmonary anastomosis, and replacement of a right ventricle to pulmonary artery homograft . CONCLUSIONS: Timely diagnosis, aggressive sternal debridement, and liberal use of rotational muscle flaps can potentially minimize the morbidity and mortality in pediatric postoperative cardiac patients . Subsequent redo-sternotomy has not been problematic.

Org Lett, 2003 Nov 13, 5(23), 4393 - 6
Unusual C25 steroids produced by a sponge-derived Penicillium citrinum; Amagata T et al.; {structure: see text} Structurally unique steroids, isocyclocitrinol A (1) and 22-acetylisocyclocitrinol A (2), were isolated from the extract of a saltwater culture of sponge-derived Penicilliun citrinum . The structures were established by analysis of 1D and 2D NMR data . The absolute structures were determined on the basis of X-ray structure analysis and application of modified Mosher's method . Furthermore, the structure of cyclocitrinol (3a) previously isolated from a terrestrial P . citrinum was revised as 3b . Compounds 1 and 2 showed weak antibacterial activity against Staphylococcus epidermidis and Enterococcus durans.

Nucl Med Rev Cent East Eur, 2002, 5(2), 93 - 7
Comparison of 99mTc-alafosfalin and 67Ga-citrate in a mouse model of bacterial infection; Tsopelas C et al.; BACKGROUND: The antibiotic-peptide (99m)Tc-alafosfalin was assessed as an infection imaging agent in Staphylococcus aureas infected mice by comparison with (99m)Ga-citrate, and also examined for influence of septic status on tracer biodistribution by comparison with normal mice . MATERIAL AND METHODS: Intramuscular doses of S . aureus were administered into the right thigh muscle of mice and the infection was allowed to develop for 20 hours . In separate experiments, (99m)Tc-alafosfalin and (67)Ga-citrate were subsequently administered and allowed to localise . Quantitative organ distribution studies were performed in conjunction with scintigraphic images at 1 and 4 hours post injection . An additional biodistribution with (99m)Tc-alafosfalin in normal mice was also performed . RESULTS: (99m)Tc-alafosfalin was predominantly renal excreted, with low liver, intestine and bone uptake . There was no difference in the uptake of these organs when infected mice were compared with normal mice . (99m)Tc-alafosfalin activity in the intestine at 1 and 4 hours was substantially less than (67)Ga-citrate . For (99m)Tc-alafosfalin, infected/non-infected thigh ratios of 2.8/1.0 and 3.6/1.0 were determined at 1 and 4 hours post injection respectively . (67)Ga-citrate gave ratios of 1.6/1.0 and 3.7/1.0 at the corresponding time points . CONCLUSIONS: (99m)Tc-alafosfalin uptake was more rapid than (67)Ga-citrate, yet diffuse at the infectious sites in mice . The small and juvenile mouse model resulted in uptake of the phosphonic acid tracer by active bone growth areas which may be a disadvantage . This (99m)Tc-antibiotic peptide has potential as an infection imaging agent, and will be investigated further in another rodent infection model in the future.

Tidsskr Nor Laegeforen, 2003 Oct 9, 123(19), 2705 - 7
{Brucellosis--a rare zoonosis}; Kittang BR et al.; BACKGROUND: Brucellosis is a rare infectious disease in Norway . MATERIAL AND METHODS: We present a patient admitted to the department of medicine at Haraldsplass Hospital, Bergen and give a review of the epidemiology, symptoms, diagnosis and treatment of Brucellosis . RESULTS AND INTERPRETATION: The clinical picture of Brucellosis is often non-specific, with swings in fever, general malaise and myalgia . Complications such as osteomyelitis, endocarditis and coagulase-negative staphylococcus (CNS) infection occur in rare cases . The diagnosis is established by detection of Brucella species in blood cultures or tissue aspirate . Brucella is often difficult to isolate, but the available serological tests are highly sensitive and specific . The infection is treated with long-term administration of a combination of two appropriate antibiotic drugs . Surgery may be necessary in case of serious complications . If adequately treated, Brucellosis has a good overall prognosis.

J Artif Organs, 2003, 6(1), 1 - 8
Recent advances in therapeutic apheresis; Bosch T; Recent advances in therapeutic apheresis include technical improvements, new indications, and pathophysiological insights . A new device that adsorbs endotoxins onto immobilized human albumin from human whole blood was recently developed . In a prospective randomized controlled trial (endotoxin adsorber study EASY), apheresis-treated patients had more improved APACHE II scores than controls . In a prospective randomized trial, the Prosorba column containing immobilized staphylococcal protein A was tested against sham apheresis in patients with end-stage rheumatoid arthritis . A significant improvement occurred in 42% of the treated patients vs . 16% of the controls . Sudden hearing loss was treated in a prospective randomized trial by a single heparin-induced extracorporeal LDL precipitation (HELP) treatment in comparison with conservative therapy . In patients with elevated fibrinogen and/or low-density lipoprotein (LDL) cholesterol levels, HELP was significantly superior to 10 days of intravenous conventional treatment . Promising results were achieved in prospective randomized trials applying immunoadsorption in end-stage dilated cardiomyopathy and rheopheresis in age-related macular degeneration . In a noncontrolled trial, C4d-positive acute humoral rejection after kidney transplantation could be effectively treated by immunoadsorption . Finally, HELP apheresis was simplified by using new hardware (HELP-Futura) . Direct adsorption of lipids (DALI)-LDL-apheresis was improved by testing DALI 1250 adsorbers with improved capacity . High-blood-flow DALI was shown to be safe and effective, with the advantage of reduced treatment time . Last but not least, a modification of dextran-sulfate cellulose LDL apheresis was developed for direct LDL hemoperfusion.

Diagn Microbiol Infect Dis, 2003 Nov, 47(3), 539 - 46
Daptomycin against multiple drug-resistant staphylococcus and enterococcus isolates in an in vitro pharmacodynamic model with simulated endocardial vegetations; Cha R et al.; Daptomycin is a lipopeptide antibiotic that exhibits bactericidal activity against Gram-positive bacteria . In pursuit of potential clinical dosing regimens for endocarditis, we evaluated two, once-daily daptomycin regimens against multiple, drug-resistant Gram-positive pathogens . Daptomycin susceptibility was determined in the absence and presence of physiologic concentrations of albumin . An in vitro pharmacodynamic model with simulated endocardial vegetations incorporating protein was used to simulate regimens of daptomycin at 6 and 8 mg/kg/day and vancomycin at 1 g every 12 h against methicillin-resistant S . aureus (MRSA-67 and 494) and S . epidermidis (MRSE-R227 and R617), glycopeptide-intermediate S . aureus and S . epidermidis (GISA-992 and GISE-12333), and vancomycin-resistant E . faecium (VREF-SF12047 and 12366) . Bacterial quantification occurred over 72 h . Daptomycin MIC results for study isolates in the absence or presence of albumin ranged from 0.125 to 4 and 1 to 8, respectively . Both daptomycin regimens achieved greater than 99.9% kill by 8 h and demonstrated greater bacterial reduction than vancomycin against all tested isolates at 24, 48, and 72 h (p < 0.05) . Undetectable limits of bacterial quantification was achieved and maintained by 8 mg/kg/day against MRSA-494 and 67, GISA-992, and VREF-590 for the study duration . Although slight regrowth was noted only for 6 mg/kg/day against MRSA-67, 99.9% kill was maintained throughout the study period without development of resistance . Pharmacodynamic profiles and drug exposure of daptomycin at 6 mg/kg/day corresponds to previously reported AUC/MIC requirements . These results suggest that 6 and 8 mg/kg/day of daptomycin represent potential regimens for further clinical evaluation in drug-resistant Gram-positive endocarditis.

Epidemiol Infect, 2003 Oct, 131(2), 931 - 7
Diversity and stability of the Staphylococcus intermedius flora in three bitches and their puppies; Saijonmaa-Koulumies LE et al.; The study investigated the transfer and the stability of the S . intermedius flora in three bitches and their puppies . A total of 240 cutaneous and mucosal isolates of S . intermedius was collected from three healthy Cavalier King Charles spaniels and their puppies during the immediate prepartum period and after whelping, over a total of 15 weeks . The isolates were genotyped with random amplified polymeric DNA-polymerase chain reaction analysis (RAPD-PCR) using two primers . Seventeen different genotypes of S . intermedius were identified . One or two of the genotypes were dominant in each of the bitches and their puppies . The rest were isolated only once or twice from the bitches or their puppies . The study indicates that S . intermedius flora within each studied bitch mainly consisted of one or two dominating and persistent clones, which were transferred from the dam to her puppies immediately after birth.

Acta Microbiol Pol, 2003, 52(2), 201 - 7
Influence of air-ions on selected microorganisms; Wiszniewski A et al.; In the following work the influence of air-ions and ozone on the selected microorganisms, first of all on Staphylococcus epidermidis is discussed . Carried out investigations proved that the levels of air-ions concentrations, causing specific changes in the development of particular microorganisms, are very differentiated and depend on individual parameters of those organisms . These levels are generally characterized by high values of concentrations, range n +/- =4 x 10(6) ions/cm3 and are connected with the type of the air-ions used . However investigations did not confirm any negative influence of the positive air-ions seen as a destructive factor upon any of the investigated microorganisms.

J Infect Dis, 2003 Nov 1, 188(9), 1284 - 94 Epub 2003 Oct 22.
Staphylococcal peptidoglycan suppresses production of interleukin-2 by T cells through a T cell-derived factor induced by direct contact between T cells and monocytes; Nakagawa Y et al.; During the last 2 decades, the incidence of sepsis due to gram-positive bacteria has increased dramatically . Nevertheless, effects of the cell-wall components that do not contain endotoxin, on immunity, are still largely unknown . Here, we demonstrate, for the first time, that the gram-positive bacterial cell-wall component peptidoglycan (PGN) severely inhibits the production of interleukin (IL)-2 by cultured human peripheral blood mononuclear cells stimulated with anti-CD3 and anti-CD28 antibodies . Furthermore, we provide evidence that the inhibitory effect is mediated predominantly by a soluble mediator produced by T cells and that the production of the inhibitory mediator is induced by direct cell-to-cell contact of T cells with PGN-stimulated monocytes . The T cell-derived inhibitory mediator is distinct from known immunosuppressive lymphokines, such as IL-10 and IL-4 . In light of the key role of IL-2 in cell-mediated immunity, it can be suggested that PGN induces the dysfunction of cell-mediated immunity.

Laryngorhinootologie, 2003 Oct, 82(10), 715 - 25; quiz 726-30
{An update on cystic fibrosis for the ENT-specialist}; Rozsasi A et al.; Patients with cystic fibrosis (CF) show a high morbidity with otorhinolaryngological disorders, especially of the nasal airways and the paranasal sinuses . Because of the underlying basic genetic defect of the respiratory mucosa in CF many patients suffer from chronic rhinosinusitis, partially combined with the development of nasal polyps . Chronic rhinosinusitis in CF is different from chronic sinusitis in Non-CF-patients . In CF mainly neutrophils are found in the nasal epithelium, whereas in Non-CF-patients eosinophils predominate . In CF-sinuses a characteristic mucosal thickening and increase of high viscous mucus is seen which is not typically found in Non-CF-patients with chronic sinusitis . The respiratory mucosa in CF is mostly colonized with Staphylococcus und Pseudomonas . These bacteria participate in the damage of the respiratory epithelia and infection of the paranasal sinuses . Repeated administration of antibiotics frequently fails to reduce subjective symptoms of the patients or polyp formations, and sinus surgery is often necessary . New therapeutic approaches, such as postoperative inhalation with Dornase-alpha, are only empiric and not frequently recommended yet . Besides the strict indication for sinus surgery, which should only be performed on an inpatient basis, the hygienic handling with CF-patients needs special attention, particularly in medical centers with many patients . An effective genetic therapy for CF is currently not available.

Rev Chir Orthop Reparatrice Appar Mot, 2003 Oct, 89(6), 479 - 86
{Ten-year surveillance of nosocomial surgical site infections in an orthopedic surgery department}; Lecuire F et al.; PURPOSE OF THE STUDY: Surveillance of nosocomial surgical site infections was instituted in our department in June 1991 . We report our first nine years experience . MATERIAL AND METHODS: This study concerned the first 9 years (June 1991-June 2000) of a surveillance program designed to monitor nosocomial surgical site infections in our orthopedic surgery department . During this period 9,696 patients underwent surgery, including 2745 for hip replacements and 1016 for knee replacements . The diagnosis of infection was based on the Centers of Disease Control criteria . Beginning in 1997, the program was widened to include all indications for prophylactic antibiotics, being limited before that time to indications for arthroplasty and spinal surgery . RESULTS: The overall rate of infection was 1.25%; 0.55% for hip arthroplasty and 1.77% for knee arthroplasty . The rate of infection among hip surgery patients over the last 5 years was much higher for prosthesis revision (2.37%) than for first-intention implantations (0.16%) . The majority of the isolated strains were Gram-positive (84%) including Staphylococcus sp . found in 65% of the cases . Multiple-strains were found in 23% of the infections . The rate of infection improved very significantly over the last five years both for knee arthroplasty and spinal surgery . The rate remained unchanged for hip arthroplasty . DISCUSSION: This 10-year survey enabled us to analyze the difficulties encountered and pinpoint errors or insufficiencies in data recording . We were also able to identify measures to be taken concerning patient follow-up . The improvement in the rate of infection over time appears to be multifactorial, undoubtedly related to wider use of prophylactic antibiotics, progress in hygiene and sterilization methods with institution of a quality assurance program, and team awareness . CONCLUSION: Surveillance of nosocomial infections is a recommended practice . We have found that the information provided can be beneficial if the data is statistically sound, pointing out the need for progress in patient follow-up.

Pneumonol Alergol Pol, 2003, 71(5-6), 253 - 60
{Sepsis with staphylococcal vegetation on tricuspid valve . Differential diagnosis with Wegener's granulomatosis}; Lotek L et al.; Acute, febrile disease in a young person with no previous medical history is most often caused by infection . If disease progresses despite antimicrobial treatment vascular and collagen diseases should be taken into account in differential diagnosis We present a case of a young man, with lung infiltrations and cavities in the course of febrile illness . He was referred to our hospital with a suspicion of Wegener's granulomatosis after few days of ineffective antimicrobial therapy . Positive blood culture for S . aureus and demonstration of tricuspid valve vegetation on echocardiographic examination enabled to make a final diagnosis of infective endocarditis with multiple lung abscessus.

Biotechnol Lett, 2003 Oct, 25(19), 1671 - 5
Selection and characterization of peptide memitopes binding to ricin; Khan AS et al.; A combinatorial random peptide display library expressed in E . coli was employed to identify short, linear peptide sequences that showed affinity for ricin and could be used as reagents for detection and identification of ricin . One peptide, P3, from a collection of four short peptides showed specific binding to ricin . The kinetic analysis of this peptide binding to the ricin showed lower equilibrium binding constants for the peptide P3 than monoclonal antibody . This is attributed due to both slower association and faster dissociation rates for the peptide P3 . The random ricin peptide P3 binds to ricin with a KD of 1 microM versus the antibody's KD of 14 nM . This particular peptide memitope P3 against ricin showed specific binding to ricin without any significant cross-reactivity against other proteins such as bovine serum albumin (BSA), lysozyme and natural bacterial toxins such as Staphylococcal enterotoxins A and B . The results provided proof-of-principal that peptide memitopes are another choice of reagents due to ease in production to be used for the detection of highly toxic bio-threat or biowarfare agents such as ricin.

J Mol Biol, 2003 Nov 7, 333(5), 893 - 905
Identification of the antigenic epitopes in staphylococcal enterotoxins A and E and design of a superantigen for human cancer therapy; Erlandsson E et al.; Monoclonal antibodies have a potential for cancer therapy that may be further improved by linking them to effector molecules such as superantigens . Tumor targeting of a superantigen leads to a powerful T cell attack against the tumour tissue . Encouraging results have been observed preclinically and in patients using the superantigen staphylococcal enterotoxin A, SEA . To further improve the concept, we have reduced the reactivity to antibodies against superantigens, which is found in all individuals . Using epitope mapping, antibody binding sites in SEA and SEE were found around their MHC class II binding sites . These epitopes were removed genetically and a large number of synthetic superantigens were produced in an iterative engineering procedure . Properties such as decreased binding to anti-SEA as well as higher selectivity to induce killing of tumour cells compared to MHC class II expressing cells, were sequentially improved . The lysine residues 79, 81, 83 and 84 are all part of major antigenic epitopes, Gln204, Lys74, Asp75 and Asn78 are important for optimal killing of tumour cells while Asp45 affects binding to MHC class II . The production properties were optimised by further engineering and a novel synthetic superantigen, SEA/E-120, was designed . It is recognised by approximately 15% of human anti-SEA antibodies and have more potent tumour cell killing properties than SEA . SEA/E-120 is likely to have a low toxicity due to its reduced capacity to mediate killing of MHC class II expressing cells . It is produced as a Fab fusion protein at approximately 35 mg/l in Escherichia coli.

Viral Immunol, 2003, 16(3), 381 - 93
Suppression of HIV-1 replication by HIV-1-irrelevant CD8+ cytotoxic T lymphocytes resulting in preservation of persistently HIV-1-infected cells in vitro; Liu H et al.; CD8+ cells of asymptomatic HIV-1 carriers (AC) contain HIV-1-specific cytotoxic T lymphocytes (CTLs) but suppress HIV-1 replication in a class I major histocompatibility complex (MHC-I)-unrestricted manner . In order to selectively investigate the HIV-1-suppressive function of CTLs apart from HIV-1-specific cytotoxicity, HIV-1-irrelevant allo-specific CTLs were established from an HIV-1-uninfected individual and their HIV-1-suppressive activity against autologous CD4+ peripheral blood mononuclear cells (PBMC) was examined . We found that these CTLs significantly suppressed both R5 and X4-HIV-1 replication in either acutely or persistently infected autologous PBMC . Although these CTLs partially killed HIV-1-infected PBMC through Fas ligand, CTLs still suppressed late steps of HIV-1 replication in the presence of neutralizing antibodies to Fas ligand . HIV-1 replication in PBMC that had been suppressed by CTLs was reversible following depletion of CTLs from culture, analogous to the previous observation for CD8+ cell-depleted PBMC of AC . Induction of HIV-1 replication by CTL-depletion was amplified by addition of newly prepared CD4+ cells or activation with staphylococcal enterotoxin B . Our results indicate that CTLs can suppress HIV-1 replication in PBMC in an antigen-nonspecific manner and preserve infected cells in a state capable of restarting HIV-1 replication and transmission.

J Immunol Methods, 2003 Oct 1, 281(1-2), 149 - 60
Affibody-beta-galactosidase immunoconjugates produced as soluble fusion proteins in the Escherichia coli cytosol; Ronnmark J et al.; Recombinant immunoconjugates constitute a novel class of immunoassay reagents produced by genetic fusion between an antigen recognizing moiety and a reporter enzyme or fluorescent protein, obviating the need for chemical coupling . In this work, we describe the construction, Escherichia coli production and characterization of recombinant beta-galactosidase (beta-gal)-based immunoconjugates directed to human immunoglobulin A (IgA) . As the antigen recognizing moieties, either monovalent or dimeric (head-to-tail) versions of an IgA-specific affibody (Z(IgA1)) were used, previously selected in vitro from a protein library based on combinatorial engineering of a single staphylococcal protein A domain . To increase the likelihood of proper presentation on the assembled homotetrameric enzyme surface, the affibody moieties were linked to the N-terminus of the enzyme subunits via a heptapeptide linker sequence . The two resulting immunoconjugates Z(IgA1)-beta-gal and (Z(IgA1))(2)-beta-gal, containing four and eight affibody moieties per enzyme, respectively, could be expressed as soluble and proteolytically stable proteins intracellularly in E . coli from where they were purified to high purity by a single anion exchange chromatography step . The yields of immunoconjugates were in the range 200-400 mg/l culture . Biosensor-binding studies showed that both the Z(IgA1)-beta-gal and (Z(IgA1))(2)-beta-gal immunoconjugates were capable of selective IgA-recognition, but with an apparent higher binding affinity for the variant containing divalent affibody moieties, presumably due to avidity effects . The applicability of this class of recombinant immunoconjugates was demonstrated by IgA detection in enzyme-linked immunosorbent assay (ELISA) and dot-blot analyses . In addition, using human kidney biopsy samples from a nephropathy patient, IgA depositions in glomeruli could be detected by immunohistochemistry with low background staining of tissue.

Eur J Immunol, 2003 Nov, 33(11), 3154 - 63
The cyclin-dependent kinase inhibitors p27Kip1 and p21Cip1 are not essential in T cell anergy; Verdoodt B et al.; Recent evidence suggests that the cyclin-dependent kinase (Cdk) inhibitors p27Kip1 and p21Cip1 are important factors in T cell anergy, but it has remained unclear whether anergy can be induced in their absence . We therefore induced anergy by stimulation of purified T cells from wild-type, p21Cip1-/-, and p27Kip1-/- mice with anti-CD3 antibodies . Anergic wild-type T cells were arrested in the G1 phase of the cell cycle with a high p27Kip1 protein level and low Cdk2 activity . In p27-/- and p21-/- T cells, the pattern of protein expression was preserved, but Cdk2 activity was increased . To confirm the in vivo relevance of these data, anergy was induced by repeated injection of mice with staphylococcal enterotoxin B (SEB), which leads to partial deletion of the responsive Vbeta8+ T cell population and anergy in the remaining T cells . p21-/- mice and wild-type mice reacted similarly to this treatment . p27-/- mice showed reduced deletion of SEB-responsive T cells, but persisting T cells were anergic . These data indicate that other cell cycle regulators contribute to the cell cycle arrest of anergic T cells, as neither Cdk inhibitor is required for the induction of anergy.

Ukr Biokhim Zh, 2003 May-Jun, 75(3), 104 - 8
{Complement-binding and immuno-modulating properties of polyreactive immunoglobulins}; Bobrovnik SA et al.; New data concerning biological properties of polyreactive immunoglobulins (PRIG) were obtained as a result of treatment of mouse serum immunoglobulins by 4 M KSCN and are presented in the paper . In particular, the capacity of PRIG to bind C1q, the subunit of the first component of complement was studied . It was shown that PRIG's binding capacity to C1q is similar to that of intact immunoglobulins . Intravenous administration of PRIG into mice together with either sheep red blood cells or heat-inactivated staphylococcal bacteria did not affect the immune response to these antigens . Meanwhile, the same administration of PRIG together with the purified protein derivate of tuberculin resulted in 10-fold increase of mouse antibody response to PPD . These results demonstrate that PRIG can have some immuno-modulating properties concerning low-immunogenic antigens.

Antimicrob Agents Chemother, 2003 Nov, 47(11), 3630 - 3
Doxycycline is anti-inflammatory and inhibits staphylococcal exotoxin-induced cytokines and chemokines; Krakauer T et al.; Proinflammatory cytokines mediate the toxic effect of superantigenic staphylococcal exotoxins (SE) . Doxycycline inhibited SE-stimulated T-cell proliferation and production of cytokines and chemokines by human peripheral blood mononuclear cells . These results suggest that the antibiotic doxycycline has anti-inflammatory effects and is therapeutically useful for mitigating the pathogenic effects of SE.

Cancer Immunol Immunother, 2004 Feb, 53(2), 118 - 24 Epub 2003 Oct 22.
In vitro biological activities of transmembrane superantigen staphylococcal enterotoxin A fusion protein; Ma W et al.; The bacterial superantigen staphylococcal enterotoxin A (SEA) stimulates T cells bearing certain TCR Vbeta domains when binding to MHC II molecules, and is a potent inducer of CTL activity and cytokine production . Antibody-targeted SEA such as C215 Fab-SEA and C242 Fab-SEA has been investigated for cancer therapy in recent years . We have previously reported significant tumor inhibition and prolonged survival time in tumor-bearing mice treated with a combination of both C215Fab-SEA and Ad IL-18 (Wang et al., Gene Therapy 8:542-550, 2001) . In order to develop SEA as an universal biological preparation in cancer therapy, we first cloned a SEA gene from S . aureus (ATCC 13565) and a transmembrane (TM) sequence from a c- erb-b2 gene derived from human ovarian cancer cell line HO-8910, then generated a TM-SEA fusion gene by using the splice overlap extension method, and constructed the recombinant expression vector pET-28a-TM-SEA . Fusion protein TM-SEA was expressed in E . coli BL21(DE3)pLysS and purified by using the histidine tag in this vector . Purified TM-SEA spontaneously associated with cell membranes as detected by flow cytometry . TM-SEA stimulated the proliferation of both human PBLs and splenocytes derived from C57BL/6 (H-2b) mice in vitro . This study thus demonstrated a novel strategy for anchoring superantigen SEA onto the surfaces of tumor cells without any genetic manipulation.

Proc Natl Acad Sci U S A, 2003 Nov 11, 100(23), 13292 - 7 Epub 2003 Oct 22.
Dynamics of the interaction between a fibronectin molecule and a living bacterium under mechanical force; Bustanji Y et al.; Fibronectin (Fn) is an important mediator of bacterial invasions and of persistent infections like that of Staphylococcus epidermis . Similar to many other types of cell-protein adhesion, the binding between Fn and S . epidermidis takes place under physiological shear rates . We investigated the dynamics of the interaction between individual living S . epidermidis cells and single Fn molecules under mechanical force by using the scanning force microscope . The mechanical strength of this interaction and the binding site in the Fn molecule were determined . The energy landscape of the binding/unbinding process was mapped, and the force spectrum and the association and dissociation rate constants of the binding pair were measured . The interaction between S . epidermidis cells and Fn molecules is compared with those of two other protein/ligand pairs known to mediate different dynamic states of adhesion of cells under a hydrodynamic flow: the firm adhesion mediated by biotin/avidin interactions, and the rolling adhesion, mediated by L-selectin/P-selectin glycoprotein ligand-1 interactions . The inner barrier in the energy landscape of the Fn case characterizes a high-energy binding mode that can sustain larger deformations and for significantly longer times than the correspondent high-strength L-selectin/P-selectin glycoprotein ligand-1 binding mode . The association kinetics of the former interaction is much slower to settle than the latter . On this basis, the observations made at the macroscopic scale by other authors of a strong lability of the bacterial adhesions mediated by Fn under high turbulent flow are rationalized at the molecular level.

J Food Prot, 2003 Oct, 66(10), 1851 - 6
Detection of staphylococcal enterotoxin B in spiked food samples; Shriver-Lake LC et al.; Contamination of food with infectious agents, intentional or not, is a global concern with far-reaching economic and social impact . Staphylococcal enterotoxins are a major cause of food poisoning, but most methods for the identification of these agents in food require extensive pretreatment or concentration of the sample prior to analysis . The array biosensor was developed as a portable device for the simultaneous analysis of multiple complex samples for multiple targets with minimal sample preparation . In this study, we use an array biosensor to expand and improve on a staphylococcal enterotoxin B (SEB) assay with the ultimate intent of incorporating testing for SEB into a battery of sensitive and convenient assays for food safety validation . In addition to buffer studies, six different types of food samples, including beverages, homogenates of fruit and meat, and carcass washings, were spiked with SEB, incubated for at least 2 h to permit antigen sequestration, and assayed . For all samples, there were differences in fluorescence intensity, but 0.5 ng of SEB per ml could be detected in <20 min with little if any pretreatment and no sample preconcentration.

Biosens Bioelectron, 2003 Nov 15, 19(2), 109 - 14
A new assay format for electrochemical immunosensors: polyelectrolyte-based separation on membrane carriers combined with detection of peroxidase activity by pH-sensitive field-effect transistor; Plekhanova YV et al.; A new rapid immunotechnique combining separation of reactants by filtration through a porous membrane and potentiometric detection of the bound enzyme label by a pH-sensitive field-effect transistor is proposed . The complexes to be detected are formed by the method described earlier in (Anal . Chem . 71 (1999) 3538), including a homogeneous binding of immunoreactants and a polyanion carrier (polymethacrylate) followed by heterogeneous separation on a membrane incorporating an immobilized polycation (poly-N-vinyl-4-ethylpyridinium) . The proposed technique for a sensitive detection of peroxidase label is based on the measurement of pH changes in the optimised substrate solution containing o-phenylenediamine, hydrogen peroxide and ascorbic acid . The antigens studied were herbicide atrazine and hormone testosterone . Their specific detection is realised via competitive binding of free and peroxidase-labelled antigens by antibodies integrating with a (staphylococcal protein A-polyanion) conjugate . The total analysis time is 20-25 min . The range of quantitative detection is 0.2-100 ng ml(-1) for atrazine and 5-300 ng ml(-1) for testosterone . Data scatter of replicate tests varies from 3 to 10% . Application of protein A-polyanion conjugate allows to use the proposed protocol for different antigens without additional treatment of specific antisera.

FEMS Microbiol Lett, 2003 Oct 10, 227(1), 47 - 51
The autolytic activity of the recombinant amidase of Staphylococcus saprophyticus is inhibited by its own recombinant GW repeats; Hell W et al.; The Aas (autolysin/adhesin of Staphylococcus saprophyticus) is a multifunctional surface protein containing two enzymatic domains an N-acetyl-muramyl-L-alanine amidase, an endo-beta-N-acetyl-D-glucosaminidase, and two different regions of repetitive sequences, an N-terminal and a C-terminal repetitive domain . The C-terminal repetitive domain is built up by the repeats R1, R2 and R3, which interconnect the putative active centers of the amidase and glucosaminidase . To investigate the influence of the C-terminal repeats and the N-terminal repeats on the amidase activity, the repetitive domains and fragments of them were cloned and expressed in Escherichia coli . The influence of the different fragments on the activity of the recombinant amidase of the Aas, consisting of the active center of the enzyme and repeat R1, was investigated in a turbidimetric microassay . The different fragments derived from the C-terminal repeats inhibited the amidase activity, while the N-terminal repeats did not influence the activity of the enzyme . The inhibiting activity increased with the number of GW repeats the recombinant fragment contained . Thus we conclude, that the C-terminal GW repeats and not the N-terminal repeats are necessary for the cell wall targeting and the autolytic function of the amidase.

Protein Eng, 2003 Sep, 16(9), 691 - 7
Inhibition of the CD28-CD80 co-stimulation signal by a CD28-binding affibody ligand developed by combinatorial protein engineering; Sandstrom K et al.; CD28 is one of the key molecules for co-stimulatory signalling in T cells . Here, novel ligands (affibodies) showing selective binding to human CD28 (hCD28) have been selected by phage display technology from a protein library constructed through combinatorial mutagenesis of a 58-residue three-helix bundle domain derived from staphylococcal protein A . Analysis of selected affibodies showed a marked sequence homology and biosensor analyses showed that all investigated affibodies bound to hCD28 with micromolar affinities (KD) . No cross-reactivity towards the related protein human CTLA-4 could be observed . This lack of cross-reactivity to hCTLA-4 suggests that the recognition site on hCD28 for the affibodies resides outside the conserved MYPPPYY motif . The apparent binding affinity for hCD28 could be improved through fusion to an Fc fragment fusion partner, resulting in a divalent presentation of the affibody ligand . For the majority of selected anti-CD28 affibodies, in co-culture experiments involving Jurkat T-cells and CHO cell lines transfected to express human CD80 (hCD80) or LFA-3 (hLFA-3) on the cell surface, respectively, pre-incubation of Jurkat cells with affibodies resulted in inhibition of IL-2 production when they were co-cultured with CHO (hCD80+) cells, but not with CHO (hLFA-3+) cells . For one affibody variant denoted Z(CD28:5) a clear concentration-dependent inhibition was seen, indicating that this affibody binds hCD28 and specifically interferes in the interaction between hCD28 and hCD80.

Mol Cell Biol, 2003 Nov, 23(21), 7638 - 47
Egr family members regulate nonlymphoid expression of Fas ligand, TRAIL, and tumor necrosis factor during immune responses; Droin NM et al.; The Fas ligand (FasL)/Fas pathway is crucial for homeostasis of the immune system and peripheral tolerance . Peripheral lymphocyte deletion involves FasL/Fas in at least two ways: coexpression of both Fas and its ligand on T cells, leading to activation-induced cell death, and expression of FasL by nonlymphoid cells, such as intestinal epithelial cells (IEC), that kill Fas-positive T cells . We demonstrate here that superantigen Staphylococcus enterotoxin B (SEB) induced a dramatic upregulation of FasL, TRAIL, and TNF mRNA expression and function in IEC from BALB/c and C57BL/6 mice . Using adoptive transfer in which CD4(+) T cells from OT-2 T-cell receptor transgenic mice were transferred into recipients, we observed an induction in IEC of FasL, TRAIL, and TNF mRNA after administration of antigen . Specific Egr-binding sites have been identified in the 5' promoter region of the FasL gene, and Egr-1, Egr-2, and Egr-3 mRNA in IEC from mice treated with SEB and from transgenic OT-2 mice after administration of antigen was upregulated . Overexpression of Egr-2 and Egr-3 induced endogenous ligand upregulation that was inhibited by overexpression of Egr-specific inhibitor Nab1 . These results support a role for Egr family members in nonlymphoid expression of FasL, TRAIL, and TNF.

J Biol Chem, 2004 Jan 9, 279(2), 1297 - 303 Epub 2003 Oct 14.
Identification of a secondary zinc-binding site in staphylococcal enterotoxin C2 . Implications for superantigen recognition; Papageorgiou AC et al.; The previously determined crystal structure of the superantigen staphylococcal enterotoxin C2 (SEC2) showed binding of a single zinc ion located between the N- and C-terminal domains . Here we present the crystal structure of SEC2 determined to 2.0 A resolution in the presence of additional zinc . The structure revealed the presence of a secondary zinc-binding site close to the major histocompatibility complex (MHC)-binding site of the toxin and some 28 A away from the primary zinc-binding site of the toxin found in previous studies . T cell stimulation assays showed that varying the concentration of zinc ions present affected the activity of the toxin and we observed that high zinc concentrations considerably inhibited T cell responses . This indicates that SEC2 may have multiple modes of interaction with the immune system that are dependent on serum zinc levels . The potential role of the secondary zinc-binding site and that of the primary one in the formation of the TCR.SEC2.MHC complex are considered, and the possibility that zinc may regulate the activity of SEC2 as a toxin facilitating different T cell responses is discussed.

Ital Heart J Suppl, 2003 Jul, 4(7), 594 - 7
{Acute embolic myocardial infarction complicating bioprosthetic stentless aortic valve endocarditis}; Brambilla G et al.; We describe the case of bioprosthetic stentless aortic valve endocarditis due to Staphylococcus epidermidis in a female patient, who previously underwent mitral and aortic valve replacement: she presented left main coronary artery occlusion due to septic embolus with subsequent myocardial infarction complicated by cardiogenic shock . We discuss the therapeutic strategies, both medical and interventional, and the surgical options.

J Am Vet Med Assoc, 2003 Oct 1, 223(7), 1013 - 6, 986
Psoriasiform-lichenoid-like dermatosis in three dogs treated with microemulsified cyclosporine A; Werner AH; Cyclosporine has been reported to be effective for the treatment of various cutaneous autoimmune disorders in dogs . Adverse reactions have generally been limited to gastrointestinal tract disturbances and cutaneous eruptions . The article describes antimicrobial-responsive cutaneous reactions in 3 dogs being treated with microemulsified cyclosporine A because of various dermatologic conditions . Cutaneous reactions in these dogs were similar to psoriasiform-lichenoid dermatitis and may represent an atypical staphylococcal infection.

Biochem Biophys Res Commun, 2003 Oct 24, 310(3), 730 - 4
Intracellular delivery of antibodies using TAT fusion protein A; Mie M et al.; Internalization of antibodies into mammalian cells is a useful method for analyzing and regulating cellular function . In this study, we developed a novel method for the delivery of antibodies into cells using the TAT-fused protein . This fusion protein consists of two functional domains, the protein transduction domain of HIV-1 TAT and the B domain of staphylococcal protein A (SpA), which has an ability to bind to the IgG . The TAT-SpA fusion protein was mixed with fluorescence-labeled rabbit IgG and added to cells . The internalization of antibody was analyzed using confocal microscopy and flow cytometry in living cells . As a result, fluorescence-labeled IgG with the TAT-SpA fusion protein was observed intracellularly . Flow cytometry results demonstrated time course and dose dependence relationships of antibody internalization . These results suggest that the TAT-SpA fusion protein can be a useful reagent for the delivery of antibody into cells.

Biotechnol Adv, 1987, 5(1), 101 - 27
Metal resistance and accumulation in bacteria; Belliveau BH et al.; Recent research on the ecology, physiology and genetics of metal resistance and accumulation in bacteria has significantly increased the basic understanding of microbiology in these areas . Research has clearly demonstrated the versatility of bacteria to cope with toxic metal ions . For example, certain strains of bacteria can efficiently efflux toxic ions such as cadmium, that normally exert an inhibitory effect on bacteria . Some bacteria such as Escherichia coli and Staphylococcus sp . can volatilize mercury via enzymatic transformations . It is also noteworthy that many of these resistance mechanisms are encoded on plasmids or transposons . By expanding the knowledge on metal-resistance and accumulation mechanisms in bacteria, it may be possible to utilize certain strains to recover precious metals such as gold and silver, or alternatively remove toxic metal ions from environments or products where their presence is undesirable.

J Lab Clin Med, 2003 Sep, 142(3), 149 - 57
Xenogeneic cells and superantigen induce human T-cell activation in the absence of T-cell recognition of xenoantigen; Diaz LA Jr et al.; The extent of interaction between human T-lymphocytes and xenogeneic antigen-presenting cells (APCs) is species-dependent . A successful interaction requires high-affinity receptor-ligand pairing across species and recognition of antigens presented by foreign major histocompatibility complex (MHC) . A deficient human T-lymphocyte response to xenogeneic cells is likely the result of a defect in these interactions . However, the requirements for a T-cell response to superantigen (SAg) may differ from those of other T-cell responses . Using irradiated murine splenocytes, which are believed to be incapable of eliciting human T-cell responses, and porcine aortic endothelial cells (PAECs) as the APC populations, we studied the human T-lymphocyte response to antigens presented by these cells . Direct proliferation of human T-lymphocytes to SAg presented by murine APCs was demonstrated; it was blocked by anti-human LFA-1 and anti-murine MHC class II but not by anti-human MHC class II . PAECs also presented SAg to human T-cells, generating a proliferative response greater than the primary response to porcine xenoantigen . Culture of human T-cells with murine splenocytes or PAECs and SAg Staphylococcus enterotoxin A (SEA) for 7 days primed human T-cells to proliferate in a secondary culture in response to autologous APCs . This autologous secondary response was human MHC class II-dependent and was inhibited by anti-human LFA-1, anti-human CD2, and anti-human CD98 . Surprisingly, both of these responses were also blocked by anti-SEA, suggesting that despite vigorous washing, a small amount of functionally important SAg was carried over from primary to secondary culture, probably bound to the surface of T-cells . Xenogeneic APCs, even those that fail to stimulate human T-cells directly, can serve as APCs for primary human T-cell responses . After such interactions T-cells can develop secondary responses in autologous interactions based on retention of minute amounts of SAg . Such interactions may have important implications for xenotransplantation.

J Med Microbiol, 2003 Nov, 52(Pt 11), 989 - 98
Evaluation of a multilocus sequence typing system for Staphylococcus epidermidis; Wang XM et al.; Staphylococcus epidermidis is a significant cause of nosocomial disease . However, the taxonomy of this pathogen, particularly at subspecies level, is unclear . A multilocus sequence typing (MLST) scheme has therefore been investigated as a tool to elucidate taxonomic relationships within this group, based on genetic relatedness . DNA sequences for internal fragments of seven housekeeping genes were compared in 47 geographically and temporally diverse S . epidermidis isolates that were obtained from clinical infections . Twenty-three different allelic profiles were detected; 17 of these were represented by single strains and the largest profile group contained 17 isolates . Diversity of the same collection of isolates was investigated by using PFGE of SmaI-digested genomic DNA to test the discrimination and validity of the MLST approach . Isolates within the largest profile group were resolved into four distinct PFGE clusters on the basis of their SmaI digest patterns . Isolates within other profile groups that contained multiple isolates had matching PFGE SmaI patterns within each group . It appears that MLST is an effective method for grouping S . epidermidis strains at the subspecies level; however, it is not as discriminatory as it has been for other species for which MLST schemes have been established and, used alone, would not be a useful method for epidemiological studies . In addition, it was demonstrated that this method was effective for confirming the identity of S . epidermidis CoNS (coagulase-negative) isolates.

Appl Environ Microbiol, 2003 Oct, 69(10), 6106 - 13
Escherichia coli ghost production by expression of lysis gene E and Staphylococcal nuclease; Haidinger W et al.; The production of bacterial ghosts from Escherichia coli is accomplished by the controlled expression of phage phiX174 lysis gene E and, in contrast to other gram-negative bacterial species, is accompanied by the rare detection of nonlysed, reproductive cells within the ghost preparation . To overcome this problem, the expression of a secondary killing gene was suggested to give rise to the complete genetic inactivation of the bacterial samples . The expression of staphylococcal nuclease A in E . coli resulted in intracellular accumulation of the protein and degradation of the host DNA into fragments shorter than 100 bp . Two expression systems for the nuclease are presented and were combined with the protein E-mediated lysis system . Under optimized conditions for the coexpression of gene E and the staphylococcal nuclease, the concentration of viable cells fell below the lower limit of detection, whereas the rates of ghost formation were not affected . With regard to the absence of reproductive cells from the ghost fractions, the reduction of viability could be determined as being at least 7 to 8 orders of magnitude . The lysis process was characterized by electrophoretic analysis and absolute quantification of the genetic material within the cells and the culture supernatant via real-time PCR . The ongoing degradation of the bacterial nucleic acids resulted in a continuous quantitative clearance of the genetic material associated with the lysing cells until the concentrations fell below the detection limits of either assay . No functional, released genetic units (genes) were detected within the supernatant during the lysis process, including nuclease expression.

Biomaterials, 2003 Nov, 24(26), 4865 - 9
Development of a long-lasting ventricular catheter impregnated with a combination of antibiotics; Kohnen W et al.; A ventricular silicone catheter impregnated with a combination of rifampin and a quinolone was developed in order to prevent ventricular shunt related infections . As model substance for the quinolones we used sparfloxacin, because of its specific physicochemical properties resulting in a quantitative detection also in the presence of a second antibiotic . In our study we focused especially on an optimization of the antibiotic release out of the impregnated catheters in order to develop long lasting devices with a broad antimicrobial spectrum . A release-optimized catheter was tested with an in vitro colonization test and additionally with a method developed to examine the spread of bacteria on a catheter surface . In vitro experiments showed that the impregnated catheters reduce the colonization with Staphylococcus epidermidis for at least 1 year and prevent the spread of bacteria along the catheter surface.

J Microbiol Methods, 2003 Nov, 55(2), 465 - 9
Evaluation of a novel method based on PCR Restriction Fragment Length Polymorphism Analysis of the tuf gene for the identification of Staphylococcus species; Kontos F et al.; A novel method, based on PCR Restriction Fragment Length Polymorphism Analysis (PRA) of a part of the tuf gene (370 bp), was designed for the identification of 11 staphylococcal species, including the most common staphylococcal pathogens . A total of 258 clinical isolates were validated by this assay, and the results were in concordance with those obtained by the reference method of Kloos and Schleifer.

Cell Immunol, 2003 Jun, 223(2), 113 - 9
Early activation events differentiate the reactivity of two T-cell families to Staphylococcus enterotoxin A; D'Souza S et al.; Analysis of early activation events in two SEA responsive T-cell families demonstrated that low doses of SEA induced CD4+Vbeta22 T-cells to down-regulate their TCR and express CD69, considerably earlier than CD4+Vbeta5 T-cells . The rapid down-regulation of Vbeta22 TCR led to its proliferation, whereas even a 10-fold higher dose of toxin induced only a partial down-regulation of Vbeta5 TCR . Stimulation with SEA induced a significantly higher percentage of Vbeta22 T-cells to produce IFN-gamma compared to Vbeta5 T-cells . SEAF47A, a mutant of SEA, known to have a lower binding affinity for the MHC class II molecule, failed to activate Vbeta5 T-cells whereas Vbeta22 T-cell activation was slightly decreased . Hence, early activation events highlighted the differential requirements of T-cell families to respond to SEA.

Nat Prod Res, 2003 Oct, 17(5), 313 - 7
Stereospecific hydroxylation of (+)-cedrol by using human skin microbial flora Staphylococcus epidermidis; Miyazawa M et al.; Microbial transformation of (+)-cedrol was investigated by using Staphylococcus epidermidis and found that stereospecific hydroxylation of (+)-cedrol occurred at the C-3 position to form (+)-(3S)-3-hydroxycedrol.

Otolaryngol Pol, 2003, 57(3), 361 - 4
{Nasal mucosa in patients with diabetes mellitus}; Muller M et al.; Diabetes mellitus is the most common endocrinologic disease all over the world . 150 million people suffer from this disease, in Poland about 2 million . The disease on the basis of the onset and pathophysiology may be divided into type I and type II . Pathophysiologic changes include diabetic microangiopathy, macroangiopathy and neuropathy . The most common presentations in head and neck are otitis externa, hypoacusis, vertigo, disequilibrium, xerostomia, dysphagia, fungal and recurrent infections . The changes in nasal mucosa are not very well known . Only few papers concerned the problem . The main complaints of patients regarding the nose are xeromycteria, hyposmia and various degree of decreased patency of the nose . Chronic atrophic rhinitis, septal perforation, ulceration of nasal mucosa, alar necrosis, symptoms of staphylococcal or fungal infection can be found during otolaryngologic examination . The treatment in this group of patients should consist of systemic therapy of diabetes mellitus and on the other hand focal therapy with the use of a solution to moisten the nasal mucosa.

Vet Immunol Immunopathol, 2003 Nov 15, 96(1-2), 13 - 8
The porcine skin associated T-cell homing chemokine CCL27: molecular cloning and mRNA expression in piglets infected experimentally with Staphylococcus hyicus; Johnsen CK et al.; CCL27 (also named CTACK, ALP, ILC and ESkine) is a CC chemokine primarily expressed by keratinocytes of the skin . The cognate receptor of CCL27 named CCR10 (GPR-2), is also expressed in skin-derived cells, and in addition by a subset of peripheral blood T-cells and in a variety of other tissues . In this paper, we report the cloning of porcine CCL27 cDNA and investigation of CCL27 mRNA expression in Staphylococcus hyicus infected piglets . At the protein level, 77 and 74% homology was found to human and mouse CCL27 sequences, respectively . The results of the expression analyses show that CCL27 mRNA is upregulated in the skin of infected piglets and to a lesser extent in piglets recovered from disease and without clinical signs of infection, indicating a role for CCL27 both during inflammation and after recovery from an infection.

Kardiol Pol, 2003 Apr, 58(4), 299 - 301
{Management of purulent mediastinitis with sternal dehiscence - a case report}; Domaradzki W et al.; The authors report the case of 70-year-old female who underwent urgent CABG for unstable angina . The post-operative course was complicated by Staphylococcal mediastinitis, which was treated with chest irrigation system and antibiotics . Because of extensive tissue destruction caused by the infection, the steel sternal wires were not placed . Interestingly, one year after surgery the sternal wound was completely healed and there were no paradoxical respiratory movements.

Am J Ophthalmol, 2003 Oct, 136(4), 649 - 54
Endophthalmitis after ganciclovir implant in patients with AIDS and cytomegalovirus retinitis; Shane TS et al.; PURPOSE: To describe the incidence, clinical course, and outcomes of endophthalmitis in patients treated with a ganciclovir implant . DESIGN: Retrospective review of cases identified from a clinical survey . METHODS: Cases of endophthalmitis were identified from a survey of 30 clinical practices known to treat a large number of patients with cytomegalovirus retinitis . RESULTS: Twenty-four cases of endophthalmitis were identified from 5185 implants performed (incidence, 0.46%) . Final visual acuity ranged from 20/20 to no light perception, with 50% (12/24) seeing light perception or worse . Vitreous cultures revealed Staphylococcus epidermidis in 29% (7/24) of patients, whereas other gram-positive organisms were responsible in 42% (10/24) of cases . In the remaining 29% (7/24) of cases, no organisms were cultured; these cases exhibited a better visual outcome (P =.007) . Six of the eight late-onset cases were associated with wound problems such as implant strut or suture exposure . There was no significant relationship between implant removal and final visual acuity . CONCLUSIONS: Endophthalmitis is an infrequent but potentially devastating complication associated with placement of a ganciclovir implant.

Acta Vet Hung, 2003, 51(3), 409 - 24
Energy imbalance related predisposition to mastitis in group-fed high-producing postpartum dairy cows; Janosi S et al.; The energy imbalance related predisposition to mastitis was studied in group-fed postpartum dairy cows (n = 333) kept in 4 large-scale units and producing milk of low somatic cell count (SCC) . Blood samples were taken on Days 1-3 after calving for assaying some metabolites and hormones related to the negative energy balance (NEB) . If mastitis was diagnosed later, aseptic milk samples were taken to identify the pathogens . Considering pathogen types {contagious pathogens: Staphylococcus (S.) aureus, Gram-positive (GP) environmental pathogens, and Gram-negative (GN) environmental pathogens + mastitis with no detectable pathogens (NDP)} separately, stepwise logistic regression was used to analyse the relation between the potential prognostic value of hormones and metabolites and mastitis outbreak . Only the elevated (> or = 1.00 mmol/l) serum beta-hydroxybutyrate (BHB) levels predisposed the cows to mastitis in the subsequent 4 weeks . This prognostic value of BHB was significant only in GN + NDP mastitis and in cases caused by GP environmental pathogens, but not in S . aureus mastitis (odds ratio: 5.333, 3.600 and 1.333, respectively).

J Biol Chem, 2003 Dec 12, 278(50), 50412 - 21 Epub 2003 Sep 27.
Dissecting cooperative and additive binding energetics in the affinity maturation pathway of a protein-protein interface; Yang J et al.; When two proteins associate they form a molecular interface that is a structural and energetic mosaic . Within such interfaces, individual amino acid residues contribute distinct binding energies to the complex . In combination, these energies are not necessarily additive, and significant positive or negative cooperative effects often exist . The basis of reliable algorithms to predict the specificities and energies of protein-protein interactions depends critically on a quantitative understanding of this cooperativity . We have used a model protein-protein system defined by an affinity maturation pathway, comprising variants of a T cell receptor Vbeta domain that exhibit an overall affinity range of approximately 1500-fold for binding to the superantigen staphylococcal enterotoxin C3, in order to dissect the cooperative and additive energetic contributions of residues within an interface . This molecular interaction has been well characterized previously both structurally, by x-ray crystallographic analysis, and energetically, by scanning alanine mutagenesis . Through analysis of group and individual maturation and reversion mutations using surface plasmon resonance spectroscopy, we have identified energetically important interfacial residues, determined their cooperative and additive energetic properties, and elucidated the kinetic and thermodynamic bases for molecular evolution in this system . The summation of the binding free energy changes associated with the individual mutations that define this affinity maturation pathway is greater than that of the fully matured variant, even though the affinity gap between the end point variants is relatively large . Two mutations in particular, both located in the complementarity determining region 2 loop of the Vbeta domain, exhibit negative cooperativity.

Nature, 2003 Sep 25, 425(6956), 411 - 4
A micrococcal nuclease homologue in RNAi effector complexes; Caudy AA et al.; RNA interference (RNAi) regulates gene expression by the cleavage of messenger RNA, by mRNA degradation and by preventing protein synthesis . These effects are mediated by a ribonucleoprotein complex known as RISC (RNA-induced silencing complex) . We have previously identified four Drosophila components (short interfering RNAs, Argonaute 2 (ref . 2), VIG and FXR) of a RISC enzyme that degrades specific mRNAs in response to a double-stranded-RNA trigger . Here we show that Tudor-SN (tudor staphylococcal nuclease)--a protein containing five staphylococcal/micrococcal nuclease domains and a tudor domain--is a component of the RISC enzyme in Caenorhabditis elegans, Drosophila and mammals . Although Tudor-SN contains non-canonical active-site sequences, we show that purified Tudor-SN exhibits nuclease activity similar to that of other staphylococcal nucleases . Notably, both purified Tudor-SN and RISC are inhibited by a specific competitive inhibitor of micrococcal nuclease . Tudor-SN is the first RISC subunit to be identified that contains a recognizable nuclease domain, and could therefore contribute to the RNA degradation observed in RNAi.

J Am Chem Soc, 2003 Oct 1, 125(39), 11988 - 92
Probing residual interactions in unfolded protein states using NMR spin relaxation techniques: an application to delta131delta; Choy WY et al.; Residual interactions in delta131delta, a large disordered fragment of staphylococcal nuclease, have been probed at two different pHs using backbone (15)N and side-chain methyl (2)H NMR spin relaxation techniques . The amplitudes of picosecond time-scale motions of both the backbone and side chains do not change considerably at either pH value, although they are significantly larger than those observed for folded proteins . In contrast, dramatic increases in the amplitudes of motions occurring on a nanosecond time scale are observed throughout delta131delta at pH 3 relative to pH 5 . This is consistent with a picture in which residual hydrophobic contacts at pH 5 are disrupted by electrostatic repulsions that dominate at the lower pH.

J Trauma, 2003 Sep, 55(3), 546 - 50
Antithrombin modulates the leukocyte-endothelial cell interaction in the staphylococcal enterotoxin B-challenged mouse; Iba T et al.; BACKGROUND: The beneficial effects of antithrombin on endotoxemia are well known . The purpose of this study was to examine the effects of antithrombin in a supertoxin-induced sepsis . METHODS: Mice were injected with staphylococcal enterotoxin B simultaneously with antithrombin . At 1 hour after injection, the mesenteric microcirculation was observed under intravital microscopy . In addition, humoral mediators were measured at the same time . RESULTS: The number of rolling leukocytes on the endothelium was significantly reduced in the treated mice (p < 0.01) . The decrease of white blood cell and platelet counts was significantly inhibited in the treated animals (p < 0.01 for both) . A comparison of the intercellular adhesion molecule-1 (p < 0.05), soluble tumor necrosis factor-alpha receptor (p < 0.05), and interleukin-6 (p < 0.01) levels showed less increase in the treated mice . CONCLUSION: Antithrombin showed a protective effects on the microcirculation of staphylococcal enterotoxin B-challenged mice by attenuating leukocyte-endothelial cell interaction . Suppression of adhesive molecule expression and cytokine production appears to play roles in this effect.

J Immunol, 2003 Oct 1, 171(7), 3475 - 84
CD4+CD25+ and CD4+CD25- T cells act respectively as inducer and effector T suppressor cells in superantigen-induced tolerance; Feunou P et al.; The repeated injection of low doses of bacterial superantigens (SAg) is known to induce specific T cell unresponsiveness . We show in this study that the spleen of BALB/c mice receiving chronically, staphylococcal enterotoxin B (SEB) contains SEB-specific CD4(+) TCRBV8(+) T cells exerting an immune regulatory function on SEB-specific primary T cell responses . Suppression affects IL-2 and IFN-gamma secretion as well as proliferation of T cells . However, the suppressor cells differ from the natural CD4(+) T regulatory cells, described recently in human and mouse, because they do not express cell surface CD25 . They are CD152 (CTLA-4)-negative and their regulatory activity is not associated with expression of the NF Foxp3 . By contrast, after repeated SEB injection, CD4(+)CD25(+) splenocytes were heterogenous and contained both effector as well as regulatory cells . In vivo, CD4(+)CD25(-) T regulatory cells prevented SEB-induced death independently of CD4(+)CD25(+) T cells . Nevertheless, SEB-induced tolerance could not be achieved in thymectomized CD25(+) cell-depleted mice because repeated injection of SEB did not avert lethal toxic shock in these animals . Collectively, these data demonstrate that, whereas CD4(+)CD25(+) T regulatory cells are required for the induction of SAg-induced tolerance, CD4(+)CD25(-) T cells exert their regulatory activity at the maintenance stage of SAg-specific unresponsiveness.

J Mol Biol, 2003 Oct 3, 332(5), 1143 - 53
Multiple parallel-pathway folding of proline-free Staphylococcal nuclease; Kamagata K et al.; When a protein exhibits complex kinetics of refolding, we often ascribe the complexity to slow isomerization events in the denatured protein, such as cis/trans isomerization of peptidyl prolyl bonds . Does the complex folding kinetics arise only from this well-known reason? Here, we have investigated the refolding of a proline-free variant of staphylococcal nuclease by stopped-flow, double-jump techniques, to examine the folding reactions without the slow prolyl isomerizations . As a result, the protein folds into the native state along at least two accessible parallel pathways, starting from a macroscopically single denatured-state ensemble . The presence of intermediates on the individual folding pathways has revealed the existence of multiple parallel pathways, and is characterized by multi-exponential folding kinetics with a lag phase . Therefore, a "single" amino acid sequence can fold along the multiple parallel pathways . This observation in staphylococcal nuclease suggests that the multiple folding may be more general than we have expected, because the multiple parallel-pathway folding cannot be excluded from proteins that show simpler kinetics.

Clin Immunol, 2003 Sep, 108(3), 182 - 9
In vitro and in vivo T cell oligoclonality following chronic stimulation with staphylococcal superantigens; Kim KS et al.; Microbial superantigens (SAg), including SEB and TSST-1, polyclonally activate T cells belonging to specific TCR BV families . A pathogenic role for SAg in various human diseases has been suggested, but enthusiasm for this view has been tempered by the T cell oligoclonality in these disorders . To assess whether T cell oligoclonality can emerge following protracted SAg stimulation, human PBMC were stimulated with SEB, TSST-1, or anti-CD3 mAb and maintained in culture with exogenous IL-2 . Oligoclonality was appreciated by day 14 among CD4(+) and CD8(+) T cells . In addition, mice transgenic for human DR2 and DQ8 were injected weekly with SEB, and splenic CD4(+) and CD8(+) T cells were analyzed for oligoclonality . In mice that received one or three such injections, little-to-no oligoclonality was detected . In contrast, considerable oligoclonality was detected in mice that received eight weekly SEB injections . Many of these T cell oligoclones were identical to "spontaneously" arising oligoclones detected in SEB-naive mice . Thus, T cell oligoclonality can emerge following chronic SAg stimulation . In hosts who have lost tolerance to self Ag, chronic exposure to SAg may preferentially promote expansion of autoreactive T cells and facilitate development of clinical disease.

Mol Biotechnol, 2003 Sep, 25(1), 89 - 94
A novel method for modification of tumor cells with bacterial superantigen with a heterobifunctional cross-linking agent in immunotherapy of cancer; Shimizu M et al.; Bacterial superantigens (SAGs) bind to cognate Vbeta elements of T-cell receptors on T-cells and to major histocompatibility complex (MHC) class II molecules on antigen-presenting cells to activate T-cell subsets expressing the Vbeta elements . We examined the possibility that the direct binding of SAGs (staphylococcal enterotoxins B {SEB} and A {SEA}) to tumor cells decreases the toxicity of SAGs, and that antitumor immunity can be induced with the aid of T-helper-1 (Th1)-type cytokines and monokines released from T-cells and monocytes, respectively, by activation with SAGs . In this context, we have developed a general method for conjugating SEB and SEA directly to tumor cells with a heterobifunctional cross linking agent, N-(gamma- maleimidobutyryloxy) sulfosuccinimide sodium salt . Using this method, we have succeeded in conjugating SEB to a sufficient extent as to induce strong tumor immunity . Both in vitro T-cell culture with SEB-bearing Meth A cells and in vivo immunization with SEB-bearing Meth A cells induce strong antitumor activity . These results suggest that the direct conjugation of SAGs including SEB and SEA to tumor cells is a powerful and useful method for immunotherapy of cancer.

Indian J Exp Biol, 2002 Nov, 40(11), 1280 - 4
Staphylococcus warneri BW 94--a new source of lipase; Walavalkar GS et al.; Staphylococcus isolated from a common Indian sweet viz . basundi was tested for its ability to produce lipase . The colorless zone of hydrolysis around the colony grown on Baird Parker agar containing egg yolk produced extracellular lipase . Colony morphology, coagulase production, haemolysis, acid production in carbohydrate medium and enzyme activity studies showed that the organism was Staphylococcus warneri . Growth of S . warneri was obtained after 11 hr at 37 degrees C, pH 7.5, while the maximum production of lipase was obtained at 30 degrees C at pH 6.5 after 9 hr of incubation . Agitation did not increase lipase production . A sudden fall in the activity of lipase was noted after 11 hr . Addition of sucrose which is a growth stimulant for Staphylococcus, did not stimulate production of lipase by these organisms . Also, addition of oleic acid, Tween 80 or ethanol did not stimulate formation of lipase.

J Huazhong Univ Sci Technolog Med Sci, 2003, 23(2), 161 - 2
Effect and mechanism of superantigen staphylococcal enterotoxin therapy for mouse gastric tumor; Li Y et al.; The anti-tumor effect and mechanism of the staphylococcal enterotoxin A (SEA) were studied . The mouse gastric tumor model was produced by subcutaneously inoculating gastric tumor cells (MGC80-3) . The experimental group was treated with SEA, and the control group was treated with normal saline . The percentage of tumor generation and tumor mass was measured . The results showed that the percentage of the tumor generation in the SEA-treated mice was lower than in the control group, but there was no significant difference (P > 0.05) . However, the tumor mass in the experimental group was significantly lighter than in the control group, with the difference being very significant (P < 0.001) . There were more CD4+ T cells and CD8+ T cells in the tumor of the mice treated with SEA than those of the control group . SEA has an obvious anti-tumor effect on mice gastric tumor . The mechanism might be that SEA induces the effect of superantigen-dependent cell mediated cytotoxicity to the tumor cells.

J Mol Biol, 2003 Sep 26, 332(4), 927 - 36
Rearrangement of charge-charge interactions in variant ubiquitins as detected by double-mutant cycles and NMR; Sundd M et al.; Previous studies of ubiquitin disclosed numerous charge-charge interactions on the protein's surface . To investigate how neighboring residues influence the strength of these interactions, double-mutant cycles are combined with pK(a) determinations by 2D NMR . More specifically, the environment around the Asp21-Lys29 ion pair has been altered through mutations at position 25, which is an asparagine in mammalian ubiquitin and a positively-charged residue in many other ubiquitin-like proteins . The pK(a) value of Asp21 decreases by 0.4 to 0.7 pH unit when Asn25 is substituted with a positively charged residue, suggesting a new and favorable ion pair interaction between positions 21 and 25 . However, analysis of double mutants reveals that the favorable interaction between Asp21 and Lys29 is weakened when position 25 is a positively charged residue . Interestingly, while the pK(a) value of His25 in the N25H variant agrees with model compound values, additional mutants reveal that this agreement is fortuitous, resulting from a balance of favorable and unfavorable interactions; similar results were observed previously for Glu34 in ubiquitin and His8 in staphylococcal nuclease . Ionizable groups may thus have pK(a) values similar to model compound values and yet still be involved in significant interactions with other protein groups . One surprising result of introducing positively charged residues at position 25 is a new interaction between Lys29 and Glu18, an interaction not present in wild-type ubiquitin . This unanticipated result illustrates a key advantage of using NMR to determine pK(a) values for many residues simultaneously in the variant proteins . Overall, the strength of an interaction between two residues at the surface of ubiquitin is sensitive to the identity of neighboring residues . The results also demonstrate that relatively conservative and common point mutations such as substitutions of polar with charged residues and vice versa can have effects on interactions beyond the site of mutation per se.

J Comp Neurol, 2003 Oct 20, 465(3), 417 - 30
Constitutive expression of p55TNFR mRNA and mitogen-specific up-regulation of TNF alpha and p75TNFR mRNA in mouse brain; Bette M et al.; Serum tumor necrosis factor (TNF) functions as a mediator of the immune-to-brain axis . Numerous TNF receptor-mediated effects on the nervous system are described but the knowledge about the regional and cellular expression of TNF receptor p55TNFR and p75TNFR in vivo is far from being complete . It is unclear whether TNF mediates its neuroimmune effects alone or in combination with other factors, e.g., bacterial mitogens . Here, we investigated the distribution of TNFalpha, p55TNFR, and p75TNFR in normal mouse brain and examined the stimulus-specific effects of lipopolysaccharide (LPS) and staphylococcal enterotoxin B (SEB) on the expression of the cerebral TNF system . Both mitogens caused enhanced TNFalpha serum levels and induced c-fos mRNA in the paraventricular nucleus but exhibited different effects on the cerebral gene expression of the TNF system . LPS but not SEB rapidly induced TNFalpha mRNA in circumventricular organs (CVOs) followed by spreading of TNFalpha mRNA into brain parenchyma close to the CVOs . The p55TNFR gene was constitutively expressed in many neurons with high levels in brainstem motor nuclei and in neurons of the sensory mesencephalic trigeminal nucleus . Moderate levels of p75TNFR mRNA were seen in single cells scattered throughout the brain in a pattern resembling microglia . Neither LPS nor SEB modulated the p55TNFR gene expression in any region or cell type of the brain, and LPS but not SEB induced p75TNFR mRNA in the CVOs . Thus, enhanced TNF serum levels able to stimulate c-fos mRNA expression in the paraventricular nucleus did not necessarily result in a modulation of the cerebral TNF system .

Appl Environ Microbiol, 2003 Sep, 69(9), 5212 - 5
Detection of Staphylococcal enterotoxin B via biomolecular interaction analysis mass spectrometry; Nedelkov D et al.; Detection of Staphylococcus enterotoxin B (SEB) by biomolecular interaction analysis mass spectrometry (BIA/MS) is presented in this work . The BIA/MS experiments were based on a surface plasmon resonance (SPR) MS immunoassay that detects affinity-captured SEB both via SPR and by means of exact and direct mass measurement by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry . Experiments were performed with standard samples and food samples to assess the BIA/MS limit of detection for SEB and to set the experimental parameters for proper quantitation . Single and double SPR referencing was performed to accurately estimate the amount of the bound toxin . Reproducible detection of 1 ng of SEB per ml, corresponding to affinity capture and MS analysis of approximately 500 amol of SEB, was readily achieved from both the standard and mushroom samples . A certain amount of SEB degradation was indicated by the signals in the mass spectra . The combination of MS with SPR-based methods of detection creates a unique approach capable of quantifying and qualitatively analyzing protein toxins from pathogenic organisms.

Acta Otolaryngol, 2003 Aug, 123(6), 704 - 8
Modulation of T-cell functions by laser surgery in patients with allergic rhinitis; Mori K et al.; OBJECTIVE: Potassium-titanyl-phosphate (KTP) laser turbinectomy is an established treatment for hypertrophied inferior turbinates that do not respond to other medical treatments . KTP laser surgery is usually performed with the aim of reducing the size of hypertrophied inferior turbinates . We hypothesized that laser vaporation may also inhibit the allergic reaction in the nasal mucosa of inferior turbinates . MATERIAL AND METHODS: We examined the effect of KTP laser therapy on T-cell responses using peripheral blood mononuclear cells (PBMCs) . RESULTS: Levels of T-cell proliferation after stimulation with Staphylococcus enterotoxin B (SEB) were higher in post- than pre-surgery patients . Levels of interferon-gamma and IL-2 produced by PBMCs after stimulation with SEB appeared to be higher in post- than pre-surgery patients . Serum levels of house dust-specific IgE were lower in post- than pre-surgery patients . These results indicate that KTP laser therapy modulates T-cell responses and probably tilts the Th1/Th2 balance towards the Th1-dominant state . CONCLUSION: KTP laser surgery eases or cures allergic rhinitis not only by reducing the volume of the inferior turbinates but also by modulating T-cell functions.

J Antimicrob Chemother, 2003 Oct, 52(4), 724 - 6 Epub 2003 Sep 01.
Prophylactic efficacy of linezolid alone or combined with levofloxacin and vancomycin in a rat subcutaneous pouch model of graft infection caused by Staphylococcus epidermidis with intermediate resistance to glycopeptides; Giacometti A et al.; OBJECTIVES: A rat model was used to investigate the efficacy of linezolid, alone or in combination with levofloxacin and vancomycin, in the prevention of vascular prosthetic graft infection resulting from methicillin-resistant Staphylococcus epidermidis with intermediate resistance to glycopeptides . METHODS: Graft infections were established in the subcutaneous tissue of adult male Wistar rats by implantation of Dacron prostheses, followed by topical inoculation with S . epidermidis . The study comprised: one group without inoculation; one inoculated group without prophylaxis; six inoculated groups that received intraperitoneal linezolid (8 mg/kg), levofloxacin (7 mg/kg) or vancomycin (7 mg/kg) alone or in combination at the dosages mentioned above . Each group included 20 animals . The grafts were removed after 7 days and evaluated by quantitative culture . RESULTS: Quantitative graft cultures from animals treated with a single drug showed a significant efficacy only for linezolid . The efficacy of levofloxacin was similar to that of vancomycin . Combination studies demonstrated that only the treatments that included linezolid produced no evidence of staphylococcal infection . CONCLUSIONS: Linezolid as perioperative prophylaxis can be useful for the prevention of graft infections caused by multiresistant staphylococcal strains.

Ai Zheng, 2003 Jun, 22(6), 582 - 5
{Influence of nanoparticle-encapsulated SEA on T-cell subgroups and its antitumor effect on hepatoma}; Ye J et al.; BACKGROUND & OBJECTIVE: Staphylococcal enterotoxin A (SEA) is one of the widely researched superantigens, which is prospective in antitumor therapy . However, its application is limited due to the toxicity . This study was conducted to prepare the nanoparticle-encapsulated SEA (NSEA) and to observe its influences on the T-cell subgroups and the antitumor effects in vivo . METHODS: NSEA was prepared by the interfacial polymerization method.BALB/c mice were divided into 3 groups (each group had 12 mice) . After injection of 0.1 ml normal saline (NS I group), 0.1 ml 2 mg/L free SEA (SEA I group) and 0.1 ml 2 mg/L NSEA(NSEA I group), the changes of T-cell subgroups (CD4(+) and CD8(+)) were observed . The mice model bearing hepatocellular carcinoma H22 were injected with 0.1 ml NS(NS II group), 0.1 ml 2 mg/L free SEA(SEA II group), 0.1 ml 2 mg/L NSEA (NESA II group), then the tumor volume and the survival time were recorded . RESULTS: SEA and NSEA significantly improved the absolute number of CD4(+) and CD8(+) T cells (P< 0.01); while the proportion of CD4(+) to CD8(+) did not change (P >0.05) . The numbers of CD4(+) and CD8(+) T cells in NSEA I group reached the peaks {(8.26+/-1.46) x 10(9)/L and (5.53+/-0.91) x 10(9)/L} at 72 hours . The absolute number of CD4(+) T cells in SEA I group reached the peak of (8.61+/-1.59) x 10(9)/L at 48 hours,and the absolute number of CD8(+) T cells reached the peak of (6.05+/-1.31) x 10(9)/L at 72 hours; both of them descended to normal level at 96 hours . The inhibition rates of SEA II group and NSEA II group were 58.9% and 50% and the percentages of life span increase were 167% and 169%, respectively . CONCLUSION: NSEA and SEA could induce the activation and proliferation of T cells in vivo but could not influence the proportion of CD4(+) and CD8(+) cells in the mice . The effects of NSEA were weaker but longer than that of SEA . This study demonstrated that NSEA has the sustained release effects and prolongs the effective time.

J Biol Chem, 2003 Nov 14, 278(46), 45216 - 23 Epub 2003 Aug 27.
Pore formation by equinatoxin II, a eukaryotic protein toxin, occurs by induction of nonlamellar lipid structures; Anderluh G et al.; Pore formation in the target cell membranes is a common mechanism used by many toxins in order to kill cells . Among various described mechanisms, a toroidal pore concept was described recently in the course of action of small antimicrobial peptides . Here we provide evidence that such mechanism may be used also by larger toxins . Membrane-destabilizing effects of equinatoxin II, a sea anemone cytolysin, were studied by various biophysical techniques . 31P NMR showed an occurrence of an isotropic component when toxin was added to multilamellar vesicles and heated . This component was not observed with melittin, alpha-staphylococcal toxin, or myoglobin . It does not originate from isolated small lipid structures, since the size of the vesicles after the experiment was similar to the control without toxin . Electron microscopy shows occurrence of a honeycomb structure, previously observed only for some particular lipid mixtures . The analysis of FTIR spectra of the equinatoxin II-lipid complex showed lipid disordering that is consistent with isotropic component observed in NMR . Finally, the cation selectivity of the toxin-induced pores increased in the presence of negatively charged phosphatidic acid, indicating the presence of lipids in the conductive channel . The results are compatible with the toroidal pore concept that might be a general mechanism of pore formation for various membrane-interacting proteins or peptides.

Biophys J, 2003 Sep, 85(3), 1980 - 95
Protein in sugar films and in glycerol/water as examined by infrared spectroscopy and by the fluorescence and phosphorescence of tryptophan; Wright WW et al.; Sugars are known to stabilize proteins . This study addresses questions of the nature of sugar and proteins incorporated in solid sugar films . Infrared (IR) and Raman spectroscopy was used to examine trehalose and sucrose films and glycerol/water solvent . Proteins and indole-containing compounds that are imbedded in the sugar films were studied by IR and optical (absorption, fluorescence, and phosphorescence) spectroscopy . Water is able to move in the sugar films in the temperature range of 20-300 K as suggested by IR absorption bands of HOH bending and OH stretching modes that shift continuously with temperature . In glycerol/water these bands reflect the glass transition at approximately 160 K . The fluorescence of N-acetyl-L-tryptophanamide and tryptophan of melittin, Ca-free parvalbumin, and staphylococcal nuclease in dry trehalose/sucrose films remains broad and red-shifted over a temperature excursion of 20-300 K . In contrast, the fluorescence of these compounds in glycerol/water solvent shift to the blue as temperature decreases . The fluorescence of the buried tryptophan in Ca-bound parvalbumin in either sugar film or glycerol/water remains blue-shifted and has vibronic resolution over the entire temperature range . The red shift for fluorescence of indole groups exposed to solvent in the sugars is consistent with the motion of water molecules around the excited-state molecule that occurs even at low temperature, although the possibility of static complex formation between the excited-state molecule and water or other factors is discussed . The phosphorescence yield for protein and model indole compounds is sensitive to the matrix glass transition . Phosphorescence emission spectra are resolved and shift little in different solvents or temperature, as predicted by the small dipole moment of the excited triplet state molecule . The conclusion is that the sugar film maintains the environment present at the glass formation temperature for surface Trp and amide groups over a wide temperature excursion . In glycerol/water these groups reflect local changes in the environment as temperature changes.

Biosens Bioelectron, 2003 Oct 15, 18(12), 1479 - 83
Piezoelectric crystal immunosensor for the detection of staphylococcal enterotoxin B; Lin HC et al.; The work of fabricating a piezoelectric (PZ) immunosensor for the detection of staphylococcal enterotoxin B (SEB) is presented in this paper . Three different immobilization methods using anti-SEB antibody onto a gold electrode of the PZ have been conducted . The electrode coated with polyethyleneimine (PEI) has shown the best result . The fabricated PZ sensor can be used for SEB determination in the range of 2.5-60 microg/ml with a correlation coefficient of 0.997 . Milk samples spiked with various concentrations of SEB gave an average of 111% recovery of the toxin . The SEB assay is specific . For example the presence of staphylococcal enterotoxin A (SEA) at 40 microg/ml gave 6.44% of the signal while staphylococcal enterotoxin D (SED) appeared to give no detectable signal . After regeneration with 1.2 M NaOH, the coated crystal could be reused three times with retention of 66% of the initial signal . The crystal has also been found to be stable for 3 days when stored at 4 degrees C in a dry atmosphere without appreciable loss of activity.

Arch Biochem Biophys, 2003 Sep 15, 417(2), 159 - 64
The recovery of dipolar relaxation times from fluorescence decays as a tool to probe local dynamics in single tryptophan proteins; Mei G et al.; The dipolar relaxation process induced by the excitation of the single tryptophan residue of four proteins (staphylococcal nuclease, ribonuclease-T1, phosphofructokinase, and superoxide dismutase) has been studied by dynamic fluorescence measurements . A new algorithm taking into account the relaxation effect has been applied to the fluorescence decay function obtained by phase-shift and demodulation data . This approach only requires that fluorescence be collected through the whole emission spectrum, avoiding the time-consuming determination of the data at different emission wavelengths, as usual with time-resolved emission spectroscopy . The results nicely match those reported in the literature for staphylococcal nuclease and ribonuclease-T1, demonstrating the validity of the model . Furthermore, this new methodology provides an alternative explanation for the complex decay of phosphofructokinase and human superoxide dismutase suggesting the presence of a relaxation process even in proteins that lack a lifetime-dependent spectral shift . These findings may have important implications on the analysis of small-scale protein dynamics, since dielectric relaxation directly probes a local structural change around the excited state of tryptophan.

Antimicrob Agents Chemother, 2003 Sep, 47(9), 3012 - 4
Postantibiotic effects of daptomycin against 14 staphylococcal and pneumococcal clinical isolates; Pankuch GA et al.; Daptomycin mean staphylococcal postantibiotic effects (PAEs) were 1.1 to 6.2 h, with a mean of 2.5 h . The mean pneumococcal PAEs were 1.7 h, ranging between 1.0 and 2.5 h . The staphylococcal and pneumococcal postantibiotic sub-MIC effects at 0.4 times the MIC ranged from 3.0 to >12.0 h and 1.9 to >12.0 h, respectively.

Clin Imaging, 2003 Sep-Oct, 27(5), 358 - 62
Imaging of primary sternal osteomyelitis in a patient with Crohn's disease; Khoury J et al.; A 41-year-old man with known Crohn's disease presented with chest pain and fever . Plain film and CT of the chest were normal, but bone scan and gallium scans showed intense sternal uptake . Surgery revealed staphylococcal osteomyelitis of the sternum . Following appropriate antibiotic treatment, the patient's symptoms resolved and follow-up gallium scans became normal . This is the first report of the association of sternal osteomyelitis and Crohn's disease, although other sites have been described . Sternal osteomyelitis has a high morbidity and the index of suspicion for this condition must be high even in the face of initially negative imaging.

Asian Pac J Allergy Immunol, 2003 Mar, 21(1), 43 - 8
Intracellular production of type I and type II cytokines during HIV-1 progression in Thai patients; Onlamoon N et al.; A type I to type II cytokine switch on cells of the immune system has been suggested as a critical step in the etiology of HIV infection . In this study, type I and type II cytokine production of both CD4+ and CD8+ T cells activated by superantigen were investigated in 10 healthy donors and 39 HIV-1 infected patients . Patients were divided into 3 groups based on their CD4 count (< 200, 200-500, > 500 cells/microl) . Whole blood from each subject was activated by staphylococcal enterotoxin B (SEB) and anti-CD28 . Intracellular cytokine stainings for proinflamatory cytokine (TNF-alpha), type I cytokines (IFN-gamma and IL-2) and type II cytokines (IL-4 and IL-5) in CD4+ and CD8+ T lymphocytes were determined by flow cytometer . Type I cytokine (IFN-gamma) expression in CD4+ T cells co-expressing with CD69 were significantly increased in HIV infected patients, particularly in patients with CD4 counts < 200 and 200-500 cells/microl (means +/- S.D . of 20.7 +/- 18.7% and 10.5 +/- 5.9%, respectively) when compared with 4.8 +/- 1.8% in the normal group (p < 0.05) . But IL-2 production in both groups of patients was significantly lower than the normal (3.8 +/- 2.6% and 3.2 +/- 1.4% in patients with < 200, 200-500 cells/microl, and 5.9 +/- 1.5% in the normal group) (p < 0.05) . For type II cytokines, there was no difference in all groups of subjects when IL-4 was determined . However, IL-5 production was significantly higher in patients with a CD4 count < 200 cells/microl (0.6 +/- 0.5%) than that in the normal group (0.1 +/- 0.1%) (p < 0.005) . CD8+ T cells also showed higher IFN-gamma production in patients with a CD4 count < 200 cells/microl (11.9 +/- 4.7%) and 200-500 cells/microl (12.0 +/- 4.3%) than the normal group (5.3 +/- 2.5%) (p < 0.005) . In contrast, IL-2 production in CD8+ T cells was low in these HIV infected patients (0.3 +/- 0.2%, 0.3 +/- 0.2%, and 0.3 +/- 0.4% in patients with < 200, 200-500, and > 500 cells/microl, respectively), which was significantly different compared to the control group (1.2 +/- 0.8%) (p < 0.05) . For type II cytokines, only IL-4 production in patients with a CD4 count < 200 cells/microl (0.1 +/- 0.1%) was significantly reduced when compared to the other groups (p < 0.05) . This study shows that although HIV infection alters the production of both type I and type II cytokines, it does not induce a polarized type I or type II state in the course of HIV-1 progression in Thai patients.

Kyobu Geka, 2003 Aug, 56(9), 783 - 5
{Prosthetic valve endocarditis after aortic valve replacement with freestyle stentless xenograft}; Tanaka H et al.; A 52-year-old man underwent aortic valve replacement with freestyle stentless xenograft, using subcoronary technique for active infective endocarditis in June, 2001 . Eighteen month later he had late prosthetic valve endocarditis associated with aortic annular abscess due to Staphylococcus epidermidis infection . The abscess was debrided and gelatin-resorcin-formalin glue (GRF glue) was injected into the abscess cavity . Abscess cavity was closed with continuous running suture of 3-0 polypropylene stitches . Finally the aortic valve was replaced with ATS mechanical valve (20 mmAP) . After administration of vancomycin and gentamicin for 4 weeks, he discharged on 57th postoperative day in good condition . We strongly suggest that GRF glue is essential to close the aortic annular abscess of combined with aortic regurgitation due to active infective endocarditis.

Surg Technol Int, 2003, 11, 57 - 60
Preventing postoperative Staphylococcus infections: an update; Grady M et al.; Postoperative nosocomial infections are associated with increased cost, hospitalization, and morbidity . S . aureus is the most common organism that contributes to postoperative nosocomial infections, and causes up to 25% of these infections . The role of the nose as a reservoir for S . aureus and possible subsequent endogenous infections has been recognized for approximately 40 years . Elimination of nasal carriage of S . aureus may be another intervention aimed at reducing postoperative infections . Mupirocin, a topical antibiotic effective against Gram-positive organisms, was proved to be effective in reducing the rates of nasal colonization of S . aureus and decreased postoperative nosocomial infections due to S . aureus.

J Immunol, 2003 Sep 1, 171(5), 2453 - 60
Adenoviral infection decreases mortality from lipopolysaccharide-induced liver failure via induction of TNF-alpha tolerance; Yarovinsky TO et al.; Effects of adenoviral infection on in vivo responses to LPS mediated by TNF-alpha were evaluated in a murine model . Adenovirus-infected mice showed decreased mortality from fulminant hepatitis induced by administration of LPS or staphylococcal enterotoxin B in the presence of D-galactosamine . Importantly, TNF-alpha resistance genes within adenoviral E3 region were not required, because E1,E3-deleted vectors showed similar effects . Adenovirus-infected mice exhibited higher TNF-alpha levels after LPS stimulation, no difference in TNFR1 expression, and similar mortality from Fas-induced fulminant hepatitis . Decreased production of IL-6 and KC in response to exogenous TNF-alpha, in addition to protection from TNF-alpha, suggested that adenoviral infection results in TNF-alpha tolerance.

Nippon Ishinkin Gakkai Zasshi, 2003, 44(3), 163 - 79
{Zygomycosis: two case reports and review of reported cases in the literature in Japan}; Mori T et al.; This article reports two cases of zygomycosis and analyzes the zygomycosis cases reported in the literature in Japan . Case 1 was a 43-year-old male with malignant lymphoma who presented complications of pneumonia and cerebral bleeding, leading to his death . Autopsy findings showed pulmonary lesions were due to zygomycosis . Cerebral lesion was presumed to be due to zygomycosis without pathological examination . Case 2 was a 52-year-old male with acute lymphocytic leukemia from whom 4 sputum cultures were taken that were positive for Cunninghamella elegans . Combination therapy of itraconazole and amphotericin B (AMPH) was begun, and AMPH was changed to liposomal amphotericin B . During the neutropenic period after receiving premedication for a peripheral blood stem cell transplantation performed for his underlying disease, high fever was recognized and Staphylococcus epidermidis was isolated from the blood culture . Despite the change in antibiotics administered, pneumonia also developed as a complication, causing his death . Two hundred four cases of zygomycosis have been reported in the literature in Japan: 55 cases were rhinocerebral zygomycosis, including 29 cases with no underlying disease . A premortem diagnosis was made in 34 cases by pathological findings of operation materials or drainage samples, and 24 cases were postmortem . Pulmonary, disseminated, cardiovascular, gastrointestinal and thyroidal zygomycoses were found in 144 cases, including 66 cases with leukemia . A premortem diagnosis was made in 39 cases and 120 cases were postmortem . Prognosis of rhinocerebral type was better in operated or drainage cases, and for resected cases in all other types . Five cases with allergic zygomycosis were all alive . There were only 14 cases in which isolated fungi were identified (Cunninghamella spp . from 5 cases, Mucor spp . from 2, Rhizomucor spp . from 2, and Rhizopus spp . from 5).

Pediatr Infect Dis J, 2003 Aug, 22(8), 731 - 6
Culture-negative osteomyelitis; Floyed RL et al.; OBJECTIVE: To examine the natural history, clinical manifestations, laboratory changes and outcomes of children with culture-negative osteomyelitis and to compare findings with cases of culture-positive infection treated during the same time period . DESIGN: Retrospective review of consecutive admissions for osteomyelitis during the 4-year period 1998 through 2001, with a minimum follow-up of 1 year . SETTING: A single urban children's hospital with a large orthopedic referral service . CLINICAL AND LABORATORY MEASURES: Age, gender, predisposing factors, clinical manifestations, maximum temperature, duration of pain, bone involved, laboratory changes, results of cultures from infected bone and blood and outcome after treatment . RESULTS: A total of 85 patients fulfilled study criteria, of whom 40 were culture-negative . Compared with culture-positive cases, culture-negative osteomyelitis patients were less likely to have antecedent trauma (P = 0.0357) and overlying skin changes (P = 0.0001), duration of pain and other symptoms was longer (P = 0.0396) and skeletal residua were rare . They were also older, with this difference approaching statistical significance (P = 0.0586) . CONCLUSIONS: Children with culture-negative osteomyelitis present initially differently from culture-positive cases and can be managed as presumed staphylococcal disease with excellent long term results.

J Cell Biol, 2003 Aug 18, 162(4), 673 - 82 Epub 2003 Aug 11.
The tyrosine phosphatase CD148 is excluded from the immunologic synapse and down-regulates prolonged T cell signaling; Lin J et al.; CD148 is a receptor-like protein tyrosine phosphatase up-regulated on T cells after T cell receptor (TCR) stimulation . To examine the physiologic role of CD148 in TCR signaling, we used an inducible CD148-expressing Jurkat T cell clone . Expression of CD148 inhibits NFAT (nuclear factor of activated T cells) activation induced by soluble anti-TCR antibody, but not by antigen-presenting cells (APCs) loaded with staphylococcal enterotoxin superantigen (SAg) or immobilized anti-TCR antibody . Immunofluorescence microscopy revealed that the extracellular domain of CD148 mediates its exclusion from the immunologic synapse, sequestering it from potential substrates . Targeting of the CD148 phosphatase domain to the immunologic synapse potently inhibited NFAT activation by all means of triggering through the TCR . These data lead us to propose a model where CD148 function is regulated in part by exclusion from substrates in the immunologic synapse . Upon T cell-APC disengagement, CD148 can then access and dephosphorylate substrates to down-regulate prolongation of signaling.

J Neurosci Methods, 2003 Aug 15, 127(2), 149 - 55
A simple method for immunocytochemical staining with multiple rabbit polyclonal antibodies; Morris TJ et al.; We report a single step, simple, repeatable, rapid and reliable technique for simultaneous immunocytochemical staining with two or more rabbit polyclonal antibodies . This technique, which we have dubbed the "Pretty Poly" method, is based on conjugating the antibodies with commercially available, fluorophore-tagged Staphylococcal protein-A (SP-A) . Staining is illustrated at the calyx type presynaptic nerve terminal of the chick ciliary ganglion with antibodies directed against three nerve terminal proteins: neurofilaments of the axonal cytoskeleton, and two secretory vesicle proteins, SV2 and cysteine string protein (CSP) . Images were deblurred with an iterative deconvolution protocol . Staining with a single polyclonal antibody was bright and had a resolution approaching light microscope limit . Treatment with two different polyclonal antibodies conjugated with contrasting dye-tagged protein-A resulted in double staining without significant crossover that was fully equivalent to the standard primary/secondary technique . The same single step protocol was used to stain with all three rabbit polyclonal antibodies or to combine the technique with a standard monoclonal primary/secondary antibody stain . Thus, the Pretty Poly protocol is a highly flexible, simple and yet effective staining technique that essentially solves the problem of co-staining with multiple polyclonal rabbit antibodies.

Contemp Top Lab Anim Sci, 2003 Jul, 42(4), 49 - 52
Inoculation of Staphylococcus xylosus in SJL/J mice to determine pathogenicity; Thornton VB et al.; An experimental study was performed to investigate whether intradermal tail inoculations of Staphylococcus xylosus would result in pathologic lesions in the SJL/J strain of mice (Mus musculus) . This organism historically has been classified as a nonpathogenic, commensal bacterium associated with skin and mucous membranes and rarely implicated in infections . In this study, SJL/J mice inoculated with S . xylosus developed cutaneous tail lesions post-inoculation, and the organism was recovered from those lesions . Inoculation was accomplished by surgically inserting silk suture impregnated with the concentrated suspension of bacteria . In addition, a superficial abrasion was created adjacent to the suture, and a bacterial suspension was applied . Approximately 80% of the mice in the inoculated groups developed dermatologic lesions, compared with 0% in the control group . Mice with lesions were treated with Sulfamethoxazole-Trimethoprim in the drinking water continuously for 28 days . For the mice assigned to the treatment group, this treatment resulted in resolution of the cutaneous tail lesions.

Appl Microbiol Biotechnol, 2003 Dec, 63(3), 293 - 9 Epub 2003 Jul 31.
Overexpression of the ATP-dependent helicase RecG improves resistance to weak organic acids in Escherichia coli; Steiner P et al.; Increased resistance to several weak organic acids was conferred on Escherichia coli by overexpression of the ATP-dependent helicase RecG and, to a lesser extent, by overexpressing the helicase RuvAB . This property of helicases was identified by reproducible selection of recG-bearing clones from genomic libraries of the acetate-resistant species Acetobacter aceti and Staphylococcus capitis . We show that overexpression of RecG from both species, but also from E . coli, increased the maximum biomass concentration attained by E . coli cultures that were grown in the presence of various weak organic acids and uncouplers . Furthermore, overexpression of RecG from A . aceti significantly improved the maximum growth rates of E . coli under weak organic acid challenge . Based on the known role of RecG in DNA replication/repair, our data provide a first indication that weak organic acids negatively affect DNA replication and/or repair, and that these negative effects may be counteracted by helicase activity.

Phys Med Biol, 2003 Jul 21, 48(14), N193 - 8
Dielectrophoretic assay of bacterial resistance to antibiotics; Johari J et al.; The dielectrophoretic collection spectra of antibiotic-sensitive and antibiotic-resistant strains of Staphylococcus epidermidis have been determined . These indicate that in the absence of antibiotic treatment there is a strong similarity between the dielectric properties of sensitive and resistant strains, and that there is a significant difference between the sensitive strains before and after treatment with the antibiotic streptomycin after 24 h exposure . This method offers possibilities for the assessment of bacterial resistance to antibiotics.

Cytokine, 2003 Jun 21, 22(6), 157 - 67
Plasticity of IL-2 and IL-2 receptor chains in rat lymphoid tissues in situ after stimulation with staphylococcal enterotoxin A; Bette M et al.; Although the effects of mitogens on the synthesis of interleukin-2 (IL-2) and IL-2 receptor (IL-2r) have been described, a detailed in situ analysis of the spatio-temporal changes of the expression of the IL-2 gene and the three IL-2r components in lymphoid tissues is still missing . Therefore, we analyzed the IL-2 and IL-2r expression after a staphylococcal enterotoxin A (SEA)-induced T cell activation on a cellular and anatomical basis in the Wistar rat . SEA caused a rapid induction of IL-2 mRNA in T cells of spleen, lymph node, and thymus, followed by the appearance of high systemic IL-2 serum levels (5 ng/ml), and a significant increase of CD25 on CD4+ and CD8+ lymphocytes . The histotopographic analysis of the IL-2r chains revealed a strong upregulation of IL-2r alpha (alpha) and IL-2r beta (beta) mRNAs in similar T cell specific compartments of spleen, lymph node, and thymus as seen for IL-2 mRNA . The abundant constitutive expression of IL-2r gamma (gamma) mRNA was unaffected by SEA . The parallel upregulation of IL-2, IL-2ralpha, and beta chains in conjunction with the continuous presence of the IL-2rgamma chain predominantly in T cell regions of immune organs suggests that the biological effects of IL-2 are essentially limited to T cells, at least after superantigen stimulation.

Tissue Antigens, 2003 Aug, 62(2), 149 - 61
Expression and function of HLA-DR3 and DQ8 in transgenic mice lacking functional H2-M; Rajagopalan G et al.; H2-M or HLA-DM are non-classical class II molecules encoded by the MHC and play an important role during antigen presentation . They catalyze exchange of CLIP (Class II-associated invariant chain peptide) or other low-affinity peptides bound to class II molecules for peptides capable of more efficient binding . The phenotype of mice lacking H2-M is determined by the allotype of the MHC class II molecules expressed . In general, H2-M deficiency does not affect the surface expression of mature class II molecules . The class II molecules in such cases predominantly contain CLIP in their peptide-binding groove . In some mice strains, H2-M deficiency results in defective CD4+ T-cell development accompanied by defective responses to conventional antigens and superantigens . Even though the HLA class II molecules show similar dependency for HLA-DM for presenting antigens in vitro, their interaction in vivo is not known . By using transgenic approach we show here that DQ8 and DR3 are expressed at normal levels in H2-M-deficient mice and the CD4+ T-cell development is unaltered . However, the ability of DQ8 molecules to present peptide antigens is compromised in a H2-M-deficient state . Presentation of exogenous bacterial superantigens by both DQ8 and DR3 is unaffected in H2-M-deficient mice . Unexpectedly, Staphylococcal Enterotoxin B-induced systemic IFN-gamma production was significantly higher in H2-M-deficient DQ8/DR3 transgenic mice and these mice were susceptible to SEB-induced toxic shock at doses that are non-lethal to H2-M-sufficient counterparts.

Biophys J, 2003 Aug, 85(2), 897 - 910
Partitioning of individual flexible polymers into a nanoscopic protein pore; Movileanu L et al.; Polymer dynamics are of fundamental importance in materials science, biotechnology, and medicine . However, very little is known about the kinetics of partitioning of flexible polymer molecules into pores of nanometer dimensions . We employed electrical recording to probe the partitioning of single poly(ethylene glycol) (PEG) molecules, at concentrations near the dilute regime, into the transmembrane beta-barrel of individual protein pores formed from staphylococcal alpha-hemolysin (alphaHL) . The interactions of the alpha-hemolysin pore with the PEGs (M(w) 940-6000 Da) fell into two classes: short-duration events (tau approximately 20 micro s), approximately 85% of the total, and long-duration events (tau approximately 100 micro s), approximately 15% of the total . The association rate constants (k(on)) for both classes of events were strongly dependent on polymer mass, and values of k(on) ranged over two orders of magnitude . By contrast, the dissociation rate constants (k(off)) exhibited a weak dependence on mass, suggesting that the polymer chains are largely compacted before they enter the pore, and do not decompact to a significant extent before they exit . The values of k(on) and k(off) were used to determine partition coefficients (Pi) for the PEGs between the bulk aqueous phase and the pore lumen . The low values of Pi are in keeping with a negligible interaction between the PEG chains and the interior surface of the pore, which is independent of ionic strength . For the long events, values of Pi decrease exponentially with polymer mass, according to the scaling law of Daoud and de Gennes . For PEG molecules larger than approximately 5 kDa, Pi reached a limiting value suggesting that these PEG chains cannot fit entirely into the beta-barrel.

Arch Dermatol Res, 2003 Sep, 295(5), 183 - 9 Epub 2003 Jul 17.
Inhibitory effect of catechin against the superantigen staphylococcal enterotoxin B (SEB); Hisano M et al.; Staphylococcal superantigens (SsAgs) have gained attention as one of the factors aggravating atopic dermatitis (AD) and several potential mechanisms of AD aggravation by SsAgs have been reported . Tea catechin has been found to have many unique antimicrobiological activities such as antibacterial, antiviral, antifungal and antitoxic effects . In the present study, we investigated the inhibitory effect of the green tea catechin extract, Polyphenon, and (-)-epigallocatechin gallate (EGCg) on staphylococcal enterotoxin B (SEB) and its mechanisms of action, and we also discuss the possibility of therapeutic benefits for AD patients of tea catechin . Polyphenon inhibited the lethal toxicity of SEB and the SEB-induced production of TNF-alpha, IFN-gamma and IL-4 following its intraperitoneal administration to BALB/c mice . Although Polyphenon is composed of several isomers among which EGCg is approximately 50% of the total, we considered that most of the inhibitory effect of Polyphenon in mice could be attributed to EGCg . EGCg was immediately bound to SEB molecules and neutralized SEB in a dose- and incubation time-dependent manner without molecular weight alteration of the SEB molecule . Furthermore, EGCg inhibited SEB-induced TNF-alpha and IFN- gamma production and IL-2, IFN-gamma, IL-10 and IL-12 p40 mRNA expression in human PBMCs from normal donors in a dose-dependent manner . Inhibition of SsAg-induced T-cell activation by catechin was observed in both in vivo and in vitro studies, suggesting that catechin may be useful in the treatment of AD.

Invest Ophthalmol Vis Sci, 2003 Aug, 44(8), 3346 - 51
Rat corneal allograft survival prolonged by the superantigen staphylococcal enterotoxin B; Pan Z et al.; PURPOSE: The purpose of this study was to determine the optimal conditions for prolonging corneal allograft survival by inducing anergy with the superantigen staphylococcal enterotoxin B (SEB) . METHODS: A rat model of penetrating keratoplasty, whereby Fisher344 donor corneas are implanted into Lewis recipients, was used to evaluate the effects of SEB on inhibiting immune-mediated allograft rejection . To induce anergy, SEB was injected into the peribulbar space of Lewis rats . Furthermore, histopathology and immunofluorescent staining were used to examine the levels of infiltrating CD4(+) and CD8(+) T lymphocytes and NK1.1(+) lymphocytes . RESULTS: By administering SEB, at doses of 90 or 120 micro g/kg 7 days before and after keratoplasty, we suppressed the episode of corneal graft rejection for a median of 12 and 30 days, respectively . In contrast, rejection was observed when 30 or 60 micro g/kg of SEB was administered . After SEB injections, lymphocyte infiltration into the corneal grafts was reduced, and the expression of NK1.1(+) lymphocytes was enhanced, suggesting that anergy may be occurring . Also, there were no differences in the number of infiltrating CD4(+) and CD8(+) T lymphocytes cells between the control group and groups injected with 30 and 120 micro g/kg SEB on postoperative days 10 and 30 . CONCLUSIONS: Inducing anergy with the superantigen SEB prolonged corneal graft survival in a rat model of penetrating keratoplasty . Therefore, these results support the possibility of prolonging corneal allograft survival in a clinical setting by preventing immune-mediated rejection through the administration of the superantigen SEB.

Indian Pediatr, 2003 Jul, 40(7), 612 - 9
Clinical profile and frequency of delta f508 mutation in Indian children with cystic fibrosis; Kabra SK et al.; OBJECTIVE: To document clinical profile of cystic fibrosis (CF) in Indian children and the prevalence of delta F508 mutation in these patients . DESIGN: Observational study . Setting:Pediatric chest clinic in an urban tertiary care center in north India . PERIOD OF STUDY: July 1995 to June 2002 . METHODS: Clinical features of 120 children diagnosed as CF by quantitative pilocarpine iontophoresis were recorded . A polymerase chain reaction based test for identification of delta F 508 mutation was performed on all children . RESULTS: Out of 3500 new cases registered in Pediatric Chest Clinic during this period 120, (3.5%) children were diagnosed as CF . Origin of parents of patients traced from almost all the States of north India . Family history suggestive of CF was present in 41 (34%) and consanguinity in 19 (61%) patients . Common clinical manifestations at the time of presentation included recurrent or persistent pneumonia in 118 (98%), failure to thrive in 108 (90%), malabsorption in 96 (80%), history of meconium ileus in 10 (8%), and rectal prolapse was present in 16 (13%) . History of salt craving, salty taste on kissing and skin rashes was present in 5 patients each . 49(41%) patients were severely malnourished . Nasal polyposis was present in 5 (4%) patients . Examination of chest revealed evidence of hyperinflation in 100 (83%), kyphosis 20 (17%), crepitations 110 (92%), wheezing 40 (25%) and bronchial breathing in 20 (17%) patients . Average clinical CF scores were 51 (95%; CI 20-80) . 48 (40%) patients had a CF score of LT40 . Pseudomonas spp was cultured from respiratory secretions of 51 (42%), Staphylococcus spp in 18 (15%), Klebsiella spp in 8 (7%) and Hemophilus influenzae in 2 (2%) patients . Delta F508 mutation was positive in 45 chromosomes out of 240 tested . Patients originated from Pakistan had more frequency of delta F508 mutations . CONCLUSIONS: Cystic fibrosis does occur in Indian children; clinical features are classical . Diagnosis is often delayed and the disease is advanced in most patients at the time of diagnosis . Frequency of Delta F508 mutation is 19% i.e., less than that seen in Caucasian population . There is need to create awareness about occurrence of CF in Indian children.

Curr Pharm Des, 2003, 9(24), 1924 - 31
Nutrition and immunity: laboratory and clinical aspects; Amati L et al.; It is well known that inappropriate nutrient intake accounts for the maintenance of the immunological equilibrium, in humans and animals . Vitamins, elements, lipids, proteins and nucleic acids play an important role in the regulation of cellular and humoral immune responses since single or multiple deficits of these food components have been shown to cause immune abnormalities . For instance, in the course of protein-calorie malnutrition bacterial and/or viral infections represent the major cause of death . Ageing is characterized by a decline of many immune functions, and this process is called immunosenescence . Here, we report novel findings on the inability of superantigens to activate old CD8+, natural killer and B cells, as an expression of cell amnesia . In the elderly, this lack of activation could lead to lethal effects in the case of severe staphylococcal infections . Quite interestingly, recent findings outlined some similarities between human immune deficiency virus (HIV)-1 infection and ageing in terms of immune changes . The model of HIV-infection may be useful for the interpretation of ageing mechanisms and possible therapeutical interventions . Finally, the role of nutrition in different pathological conditions and the use of medical foods for correcting of immune deficits will be described.

J Food Prot, 2003 Jul, 66(7), 1222 - 6
Immunomagnetic flow cytometric detection of staphylococcal enterotoxin B in raw and dry milk; Miyamoto T et al.; A rapid and sensitive method for detection of staphylococcal enterotoxin B (SEB) in raw and dry milk samples with the use of antibody-based immunomagnetic separation (IMS) in conjunction with flow cytometry (FCM) was developed . Sheep anti-SEB immunoglobulin G (IgG) was immobilized on Dynabeads M-280 . The SEB initially binds to the capturing antibody, which is bound on the magnetic beads . The rabbit anti-SEB IgG binds to the captured toxin and is further labeled with a Cy5-labeled goat anti-rabbit IgG antibody . The percentage of the beads that were fluorescent was measured by FCM . FCM was carried out for 1 min, and the data obtained were expressed as histograms for particle size (forward light scatter) and histograms for fluorescence intensity . A peak corresponding to the magnetic beads was clearly distinguished from a peak derived from contaminating particles in the sample solution . In the absence of SEB, about 10% of the beads emitted fluorescence . The percentage of fluorescent beads and the fluorescence intensity increased with increasing SEB concentrations . For this IMS-FCM assay, the lower limits of detection for SEB were estimated to be 0.01 and 0.25 ng/ml for buffer and milk samples, respectively.

Proteins, 2003 Aug 15, 52(3), 360 - 99
Atoms-in-molecules study of the genetically encoded amino acids . III . Bond and atomic properties and their correlations with experiment including mutation-induced changes in protein stability and genetic coding; Matta CF et al.; This article presents a study of the molecular charge distributions of the genetically encoded amino acids (AA), one that builds on the previous determination of their equilibrium geometries and the demonstrated transferability of their common geometrical parameters . The properties of the charge distributions are characterized and given quantitative expression in terms of the bond and atomic properties determined within the quantum theory of atoms-in-molecules (QTAIM) that defines atoms and bonds in terms of the observable charge density . The properties so defined are demonstrated to be remarkably transferable, a reflection of the underlying transferability of the charge distributions of the main chain and other groups common to the AA . The use of the atomic properties in obtaining an understanding of the biological functions of the AA, whether free or bound in a polypeptide, is demonstrated by the excellent statistical correlations they yield with experimental physicochemical properties . A property of the AA side chains of particular importance is the charge separation index (CSI), a quantity previously defined as the sum of the magnitudes of the atomic charges and which measures the degree of separation of positive and negative charges in the side chain of interest . The CSI values provide a correlation with the measured free energies of transfer of capped side chain analogues, from the vapor phase to aqueous solution, yielding a linear regression equation with r2 = 0.94 . The atomic volume is defined by the van der Waals isodensity surface and it, together with the CSI, which accounts for the electrostriction of the solvent, yield a linear regression (r2 = 0.98) with the measured partial molar volumes of the AAs . The changes in free energies of transfer from octanol to water upon interchanging 153 pairs of AAs and from cyclohexane to water upon interchanging 190 pairs of AAs, were modeled using only three calculated parameters (representing electrostatic and volume contributions) yielding linear regressions with r2 values of 0.78 and 0.89, respectively . These results are a prelude to the single-site mutation-induced changes in the stabilities of two typical proteins: ubiquitin and staphylococcal nuclease . Strong quadratic correlations (r2 approximately 0.9) were obtained between DeltaCSI upon mutation and each of the two terms DeltaDeltaH and TDeltaDeltaS taken from recent and accurate differential scanning calorimetry experiments on ubiquitin . When the two terms are summed to yield DeltaDeltaG, the quadratic terms nearly cancel, and the result is a simple linear fit between DeltaDeltaG and DeltaCSI with r2 = 0.88 . As another example, the change in the stability of staphylococcal nuclease upon mutation has been fitted linearly (r2 = 0.83) to the sum of a DeltaCSI term and a term representing the change in the van der Waals volume of the side chains upon mutation . The suggested correlation of the polarity of the side chain with the second letter of the AA triplet genetic codon is given concrete expression in a classification of the side chains in terms of their CSI values and their group dipole moments . For example, all amino acids with a pyrimidine base as their second letter in mRNA possess side-chain CSI < or = 2.8 (with the exception of Cys), whereas all those with CSI > 2.8 possess an purine base . The article concludes with two proposals for measuring and predicting molecular complementarity: van der Waals complementarity expressed in terms of the van der Waals isodensity surface and Lewis complementarity expressed in terms of the local charge concentrations and depletions defined by the topology of the Laplacian of the electron density . A display of the experimentally accessible Laplacian distribution for a folded protein would offer a clear picture of the operation of the "stereochemical code" proposed as the determinant in the folding process .

Clin Immunol, 2003 Jul, 108(1), 51 - 9
Generation of protective immunity by inactivated recombinant staphylococcal enterotoxin B vaccine in nonhuman primates and identification of correlates of immunity; Boles JW et al.; At this time there are no vaccines or therapeutics to protect against staphylococcal enterotoxin B (SEB) exposure . Here, we report vaccine efficacy of an attenuated SEB in a nonhuman primate model following lethal aerosol challenge and identify several biomarkers of protective immunity . Initial in vitro results indicated that the mutation of key amino acid residues in the major histocompatibility complex (MHC) class II binding sites of SEB produced a nontoxic form of SEB, which had little to no detectable binding to MHC class II molecules, and lacked T-cell stimulatory activities . When examined in a mouse model, we found that the attenuated SEB retained antigenic structures and elicited protective immune responses against wild-type SEB challenge . Subsequently, a vaccine regimen against SEB in a nonhuman primate model was partially optimized, and investigations of immune biomarkers as indicators of protection were performed . SEB-naive rhesus monkeys were vaccinated two or three times with 5 or 20 microg of the attenuated SEB and challenged by aerosol with wild-type SEB toxin . Unlike exposure to the native toxin, the vaccine did not trigger the release of inflammatory cytokines (TNF alpha, IL6, or IFN gamma) . All rhesus monkeys that developed anti-SEB serum titers > or = 10(4) and elicited high levels of neutralizing antibody survived the aerosol challenge . These findings suggest that the attenuated SEB is fully protective against aerosolized toxin when administered to unprimed subjects . Moreover, experiments presented in this study identified various biomarkers that showed substantial promise as correlates of immunity and surrogate endpoints for assessing in vivo biological responses in primates, and possibly in humans, to vaccines against SEs.

J Vet Med B Infect Dis Vet Public Health, 2003 Jun, 50(5), 241 - 6
Differentiation of Staphylococcus sciuri strains isolated from free-living rodents and insectivores; Hauschild T et al.; Twenty-nine Staphylococcus sciuri strains isolated from free-living insectivores and rodents were comparatively analysed for their biochemical capacities and their SmaI macrorestriction patterns . The 29 S . sciuri isolates represented 21 different biotypes and 22 different SmaI macrorestriction types . This observation confirmed that S . sciuri isolates obtained from insectivores and rodents living in natural environments constituted a heterogeneous population . Cluster analysis revealed that the macrorestriction patterns and the biochemical profiles matched in some cases . However, S . sciuri isolates that exhibited the same or closely related biochemical profiles were also found to be associated with different macrorestriction patterns . Analysis of the 29 S . sciuri isolates for their plasmid content and their sensitivity to antimicrobial agents showed that most of the isolates were plasmid-free and susceptible to all antimicrobial agents tested.

J Infect, 2003 Aug, 47(2), 164 - 6
Successful use of oral linezolid as a single active agent in endocarditis unresponsive to conventional antibiotic therapy; Ravindran V et al.; Treatment of resistant gram-positive endocarditis is difficult . We report a case of resistant Staphylococcus epidermidis endocarditis that failed to respond to conventional antibiotic therapy but was treated successfully with an oral regimen of a new antibiotic, linezolid as a single active agent . This case report demonstrates the use of linezolid as an effective alternative to conventional antibiotics in such cases.

Allergy, 2003 Aug, 58(8), 767 - 71
Subjects with allergic rhinitis show signs of more severely impaired paranasal sinus functioning during viral colds than nonallergic subjects; Alho OP et al.; BACKGROUND: Viral cold is thought to be the major contributing factor in the pathogenesis of sinusitis, as it causes ostiomeatal obstruction . The aim was to evaluate whether paranasal sinus functioning during viral colds is similar in subjects with and without allergic rhinitis . METHODS: Forty-eight volunteers were examined during an early (2-4 days) natural cold and again 3 weeks later . The examinations included computed tomography (CT) scans, nasal mucosal biopsies, and viral and bacterial specimens . Subjects with positive skin prick tests and persistent or intermittent rhinitis were considered to have allergic immunoglobulin E (IgE)-mediated rhinitis . In addition, specific IgE antibodies to staphylococcal enterotoxin B (SEB) were measured . RESULTS: Nine subjects (19%) had allergic rhinitis . The allergic subjects were significantly more often IgE sensitized to SEB than the nonallergic subjects (33%vs 3%, P = 0.02) . Viral etiology of the cold was identified in 32 (67%) subjects . The subjects with allergic rhinitis had significantly higher CT scores compared with nonallergic subjects during the colds (median (range) scores 16 (6-22) vs 6 (0-17), P = 0.004) . In both groups, the median scores declined markedly during convalescence, but the difference remained significant (P = 0.009) . Among the allergic subjects, those who were IgE sensitized to SEB tended to have the highest CT scores {median (range) 16 (16-22)} . Total serum IgE and the nasal subepithelial eosinophil counts correlated with the CT scores during the cold (rs = 0.38, P = 0.008 and rs = 0.46, P = 0.001, respectively) . CONCLUSIONS: Subjects with allergic IgE-mediated rhinitis had more severe paranasal sinus changes in CT scans than nonallergic subjects during viral colds . These changes indicate impaired sinus functioning and may increase the risk of bacterial sinusitis.

Bioorg Med Chem Lett, 2003 Aug 4, 13(15), 2449 - 53
Side-chain-to-tail thiolactone peptide inhibitors of the staphylococcal quorum-sensing system; Scott RJ et al.; The expression of many staphylococcal virulence factors are regulated by the agr locus via a two-component signal transduction system (TCSTS), which is activated in response to a secreted autoinducer peptide (AIP) . By exploiting the unique chemical architecture of the naturally occurring AIP-1, several potent inhibitors of staphylococcal TCSTS were designed and synthesized using either a linear or branched solid-phase approach . These inhibitors are competitive binders and contain the crucial 16-membered side-chain-to-tail thiolactone peptide pharmacophore.

Toxicol Pathol, 2003 Jul-Aug, 31(4), 373 - 8
Murine lethal toxic shock caused by intranasal administration of staphylococcal enterotoxin B; Savransky V et al.; Currently available murine staphylococcal enterotoxin B (SEB) shock models require pretreatment with various agents to increase mouse sensitivity to SEB . This study was performed to show that C3H/HeJ mice are highly susceptible to intranasal SEB inoculation, which caused toxic shock without using pretreatment agents . For this purpose, mice were injected intranasally with different doses of SEB and observed for up to 1 month . The median lethal dose of SEB was determined using the probit procedure . Tissue samples were taken at different time points for histopathological examination . The LD(50) was found at 1.6 microg/g (95% fiducial limit (f.l.) 0.7 to 2.2), the LD(80) at 2.7 microg/g (95% f.l . 1.9 to 4.0) and the LD(90) at 3.6 microg/g (95% f.l . 2.7 to 6.4) . Histopathologic examination revealed pulmonary edema and bronchopneumonia . Mucosal-associated lymphoid tissue first became activated, followed by increasing lymphocyte apoptosis and depletion . In the liver there were intralobular and portal inflammatory foci with increasing lymphocyte apoptosis and degenerative necrosis . The splenic white pulp was characterized by early activation and subsequent depletion of lymphoid follicle germinal centers . The thymus initially was activated, followed by increasing apoptosis and migration of lymphoid cells from the cortex to the medulla . The pathological features detected in the mice were similar to those of rhesus monkeys treated with SEB aerosol challenge.

J Mol Biol, 2003 Jul 18, 330(4), 821 - 37
Native-like partially folded conformations and folding process revealed in the N-terminal large fragments of staphylococcal nuclease: a study by NMR spectroscopy; Feng Y et al.; The N-terminal large fragments of staphylococcal nuclease (SNase), SNase110 (1-110 residues), SNase121 (1-121 residues), and SNase135 (1-135 residues), and the fragment mutants G88W110, G88W121, V66W110 and V66W121 were studied by heteronuclear multidimensional NMR spectroscopy . Ensembles of co-existent native-like partially folded and unfolded states were observed for fragments . The persistent native-like tertiary interaction drives fragments to be in partially folded states, which reveal native-like beta-barrel conformations . G88W and V66W mutations modulate the extent of inherent native-like tertiary interaction in fragment molecules, and in consequence, fragment mutants fold into native-like beta-subdomain conformations . In cooperation with the inherent tertiary interaction, 2 M TMAO (trimethylamine N-oxide) can promote the folding reaction of fragments through the changes of unfolding free energy, and a native-like beta-subdomain conformation is observed when the chain length contains 135 residues . Heterogeneous partially folded conformations of 1-121 and 1-135 fragments due to cis and trans X-prolyl bond of Lys116-Pro117 make a non-unique folding pathway of fragments . The folding reaction of fragments can be characterized as a hierarchical process.

J Immunol, 2003 Jul 15, 171(2), 1102 - 8
Differential susceptibility to staphylococcal superantigen (SsAg)-induced apoptosis of CD4+ T cells from atopic dermatitis patients and healthy subjects: the inhibitory effect of IL-4 on SsAg-induced apoptosis; Lin YT et al.; This study had two aims: 1) to determine whether there are differences between atopic dermatitis (AD) patients and healthy subjects in staphylococcal superantigen (SsAg)-induced CD4(+) T cell activation, cytokine production, chemokine receptor expression, and apoptosis; and 2) to investigate the effect of IL-4 on SsAg-induced apoptosis . By using immunofluorescence and annexin V staining, we analyzed PBMC with or without staphylococcal enterotoxin B (SEB) stimulation in the presence or absence of rIL-4 or anti-IL-4-neutralizing Abs in 15 healthy subjects and 27 AD patients . We found that SEB preferentially induced production of Th1 cytokine in SEB-reactive (TCRVbeta3(+) or Vbeta12(+) or Vbeta17(+)) CD4(+) T cells from healthy subjects and Th2 cytokine in those from AD patients . SEB induced up-regulation of CXCR3(+) cells in SEB-reactive CD4(+) T cells from healthy subjects and CCR4(+) cells in those from AD patients . SEB-reactive CD4(+) T cells from AD patients were more resistant to SEB-induced apoptosis than those from healthy subjects . There was no significant difference between AD and healthy subjects in SEB-induced activation of CD4(+) T cells . CXCR3(+) CD4(+) T cells were more susceptible to SEB-induced apoptosis than CCR4(+) CD4(+) T cells in healthy subjects . Exogenously added IL-4 inhibited SEB-induced apoptosis of SEB-reactive CD4(+) and CXCR3(+) CD4(+) T cells but not of CCR4(+) CD4(+) T cells in healthy subjects . Inhibition of endogenous IL-4 increased SEB-induced apoptosis of SEB-reactive CD4(+) T cells from AD patients . These results might provide new clues to the mechanism that SsAgs contribute to the persistence and exacerbation of allergic skin inflammation in AD.

J Immunol, 2003 Jul 15, 171(2), 860 - 6
Linker for activation of T cells, zeta-associated protein-70, and Src homology 2 domain-containing leukocyte protein-76 are required for TCR-induced microtubule-organizing center polarization; Kuhne MR et al.; Engagement of the T cell with Ag on an APC results in a series of immediate signaling events emanating from the stimulation of the TCR . These events include the induced phosphorylation of a number of cellular proteins with a subsequent increase in intracellular calcium and the restructuring of the microtubule and actin cytoskeleton within the T cell . This restructuring of the cytoskeleton culminates in the polarization of the T cell's secretory apparatus toward the engaging APC, allowing the T cell to direct secretion of cytokines toward the appropriate APC . This polarization can be monitored by analyzing the position of the microtubule-organizing center (MTOC), as it moves toward the interface of the T cell and APC . The requirements for MTOC polarization were examined at a single-cell level by studying the interaction of a Jurkat cell line expressing a fluorescently labeled MTOC with Staphylococcal enterotoxin superantigen-bound Raji B cell line, which served as the APC . We found that repolarization of the MTOC substantially followed fluxes in calcium . We also used immobilized anti-TCR mAb and Jurkat signaling mutants, defective in TCR-induced calcium increases, to determine whether signaling components that are necessary for a calcium response also play a role in MTOC polarization . We found that zeta-associated protein-70 as well as its substrate adaptor proteins linker for activation of T cells and Src homology 2 domain-containing leukocyte protein-76 are required for MTOC polarization . Moreover, our studies revealed that a calcium-dependent event not requiring calcineurin or calcium/calmodulin-dependent kinase is required for TCR-induced polarization of the MTOC.

J Immunol, 2003 Jul 15, 171(2), 664 - 8
Febrile temperatures attenuate IL-1 beta release by inhibiting proteolytic processing of the proform and influence Th1/Th2 balance by favoring Th2 cytokines; Boneberg EM et al.; We investigated possible feedback mechanisms of febrile temperatures on LPS- and staphylococcal enterotoxin B (SEB)-induced cytokine release in human whole blood . LPS-induced IL-1beta release was inhibited at temperatures >38 degrees C, whereas intracellular proIL-1beta formation as well as the release of other cytokines except IL-18 were only attenuated above 42 degrees C, indicating that febrile temperatures impair the proteolytic processing of proIL-1beta . This attenuated processing is not due to either heat inactivation of caspase-1 or structural changes in proIL-1beta produced at higher temperatures . Instead, we propose that febrile conditions change cytosolic compartmentation or trafficking, so that synthesized proIL-1beta cannot encounter caspase-1 . Febrile temperatures also influenced Th1/Th2 cytokine balance . We observed a 3-fold increase in the Th2-cytokines IL-5 and IL-13 and a reduction to 15% of the Th1-cytokine IL-2 when SEB-stimulated whole blood was incubated at 40 degrees C compared with 37 degrees C . These results indicate that fever limits the production of the fever-inducing IL-1beta and also influences the adaptive immune response, favoring Th2 cytokine production.

J Clin Microbiol, 2003 Jul, 41(7), 3348 - 50
Involvement of adherence and adhesion Staphylococcus epidermidis genes in pacemaker lead-associated infections; Klug D et al.; We explored three genes of attachment (fbe and atlE) and adhesion (ica) in 27 and 10 Staphylococcus epidermidis strains involved in pacemaker-related infections (PMI) and intravascular-catheter-related infections (IVCI), respectively, and in 25 saprophytic strains . The detection rates of fbe and atlE were identical in PMI and IVCI strains, but ica detection rates were identical in PMI and saprophytic strains.

J Cataract Refract Surg, 2003 Jun, 29(6), 1211 - 8
Staphylococcus epidermidis adherence to human amniotic membrane and to human, rabbit, and cat conjunctiva; John T et al.; PURPOSE: To evaluate Staphylococcus epidermidis adherence to human amniotic membrane (HAM) and compare it with S epidermidis adherence to human, rabbit, and cat conjunctiva in vitro . SETTING: Research laboratory, Loyola University Medical Center, Maywood, Illinois, USA . METHODS: Commercially available HAM (N = 3) was used . Conjunctival specimens from humans, rabbits, and cats (n = 3 each) were processed similarly to HAM . The tissues were exposed to S epidermidis (3 x 10(8) colony-forming units per milliliter) for 0, 5, 30, and 90 minutes, rinsed in sterile saline, and processed for light, scanning (SEM), and transmission (TEM) electron microscopy . Scanning electron microscopy (x2000) was used to quantify adherent bacteria/mm(2) of tissue (SEM photographs = 144) . RESULTS: The following mean levels (+/- SD) of adherent S epidermidis/mm(2) were found at 0, 5, 30, and 90 minutes: HAM, 3833 +/- 1570, 9060 +/- 2512, 15,431 +/- 10,752, and 30,315 +/- 14,803, respectively; human conjunctiva, 1493 +/- 672, 7218 +/- 3179, 17,273 +/- 7168, and 19,861 +/- 9624, respectively; rabbit conjunctiva, 3385 +/- 5074, 14,386 +/- 14,569, 15,283 +/- 13,679, and 20,113 +/- 24,016, respectively; and cat conjunctiva, 4032 +/- 2240, 12,345 +/- 3413, 8512 +/- 4032, and 19,214 +/- 5584, respectively . No statistically significant differences were found at any time point (P>.16) . CONCLUSION: There was no statistically significant difference in the adherence of S epidermidis to HAM and to human, rabbit, and cat conjunctiva . Bacterial adherence to HAM may be clinically significant.

Zhonghua Zhong Liu Za Zhi, 2003 May, 25(3), 225 - 9
{Effect of activated greater omental milky spots and peritoneal macrophages on tumoricidal activity against gastric carcinoma SGC-7901 in mice}; Mao WZ et al.; OBJECTIVE: To investigate the effect of activated greater omental milky spots and peritoneal macrophages in mice on tumoricidal activity against gastric carcinoma SGC-7901, following intraperitoneal (i.p.) injection of INF-gamma, staphylococcin aureus or NDV-L . METHODS: The quantitative changes of milky spots were determined by activated carbon, the number of the macrophage in milky spots was assessed by nonspecific esterase stain and the number of peritoneal macrophages was counted by trypan blue exclusion . The morphology of peritoneal macrophages was observed by scanning electron microscope, the amount of TNF-alpha and iNOS mRNA expressed by peritoneal macrophages was measured by fluorescence quantitative PCR and the cytotoxicity of peritoneal macrophages supernatant against SGC-7901 was evaluated by MTT assay . RESULTS: It was found in the treated groups that: 1 . The amount of greater omental milky spots and the macrophages in milky spots increased, 2 . The number of peritoneal macrophages increased . The peritoneal macrophages were in activated status . The effect TNF-alpha and iNOS mRNA expression increased and 3 . The cytotoxicity against in vitro SGC-7901 increased . CONCLUSION: Intraperitoneal injection of IFN-gamma, staphylococcin aureus or NDV-L could activate the milky spots of the greater omentum and the macrophages in peritoneal cavity in mice, with IFN-gamma being the best . The supernatant of activated peritoneal macrophages has cytotoxicity against SGC-7901 . Administration of LPS to macrophages cultured in vitro could amplify the activation and enhance the cytotoxicity of the supernatant against SGC-7901.

Am J Orthop, 2003 Jun, 32(6), 302 - 5
Delayed-onset Mycobacterium tuberculosis infection with staphylococcal superinfection after total knee replacement; Al-Shaikh R et al.; Infection of a total joint replacement with Mycobacterium tuberculosis is uncommon in North America . This case describes a staphylococcal superinfection that masked an underlying tuberculous infection after total knee replacement and subsequent placement of a cement spacer . The patient had no evidence of M tuberculosis infection elsewhere . The most common explanation for these events is local reactivation of quiescent tuberculosis of the knee joint . The patient was treated successfully with surgical debridement, arthrodesis, and antituberculous medication.

J Biomed Mater Res A, 2003 Jul 1, 66(1), 21 - 8
The inhibitory activity of serum to prevent bacterial adhesion is mainly due to apo-transferrin; Ardehali R et al.; A marked, up to 5-fold, reduction in bacterial adhesion to Tecoflex polyurethane (PU) surfaces was observed in the presence of bovine/human serum or plasma at 0.5% or higher concentrations in the medium . Further investigation of the phenomenon resulted in identification, isolation, and characterization of the serum component with the ability to significantly reduce bacterial adhesion . Upon fractionation of bovine serum by an anion exchange chromatography, protein pools were made and analyzed by immunoelectrophoresis and by polyacrylamide gel electrophoresis in the presence of SDS and were examined for their effect on the adhesion of Staphylococcus epidermidis to PU surfaces . The pool exhibiting a significant inhibitory effect was subjected to further biochemical tests, which resulted in the identification of transferrin (Tf) as its predominant protein . Bacterial adhesion studies in the presence of purified Tf revealed that holo-Tf (iron-containing form) had no influence on bacterial adhesion at any concentration . Only apo-Tf (iron-lacking form) exerted the inhibitory effect, in a dose responsive manner at concentrations of 10 microg/mL or higher . Bacteria remained viable when suspended at the low apo-Tf concentrations, sufficient to prevent bacterial adhesion .

Transplantation, 2003 Jun 27, 75(12), 2090 - 9
Mycophenolate mofetil-based immunosuppression and cytokine genotypes: effects on monokine secretion and antigen presentation in long-term renal transplant recipients; Weimer R et al.; BACKGROUND: It has been suggested that increased monocyte responses might play a role in chronic allograft rejection . METHODS: We investigated in vitro monokine responses in 112 patients with long-term stable kidney graft function (ST patients; n=80, non-mycophenolate mofetil {MMF}; n=32, MMF) and 25 patients with chronic renal transplant rejection (CR patients; non-MMF) . Interleukin 10 and tumor necrosis factor (TNF)-alpha promoter gene polymorphisms were tested by polymerase chain reaction and sequence-specific primers; antigen-presenting capacity (AC) of monocytes was tested by incubation with staphylococcal superantigens (SEA, SEE, SED) . RESULTS: Although non-MMF-based immunosuppression in ST patients did not result in compromised AC or lipopolysaccharide (LPS)-stimulated monokine responses compared with healthy controls, we found MMF therapy to be associated with significantly reduced TNF-R1 expression on monocytes (P<0.001), suppressed AC (P<0.02, SED), and suppressed LPS-stimulated IL-1 beta, IL-10, and TNF-alpha secretion (P<0.01) . Coinciding with a significantly higher steroid dosage in CR patients, IL-6 receptor and TNF-R1 expression on monocytes were down-regulated (P< or =0.02) and AC was suppressed in CR compared with ST (non-MMF) patients (P<0.01, SED; P<0.05, SEE) . However, LPS-stimulated monokine secretion was not decreased or even enhanced (IL-6, granulocyte-macrophage colony-stimulating factor {GM-CSF}; P<0.05) . Enhanced in vitro IL-10 responses (>500 pg/mL) were found predominantly in non-MMF-treated patients with the IL-10 genotype GCC (GCC: 23/62 {37%}, non-GCC: 2/27 {7%}, P<0.005; GCC and non-MMF: 22/47 {47%}, GCC and MMF: 1/15 {7%}, P<0.005} . CONCLUSION: Steroids and azathioprine did not sufficiently suppress monokine responses, whereas MMF treatment might inhibit chronic graft rejection because of suppression of TNF-R1 expression and vigorous inhibition of monokine secretion . MMF treatment may especially be indicated in patients with the IL-10 "high-producer" genotype GCC.

Biophys J, 2003 Jul, 85(1), 108 - 25
Protein stability in mixed solvents: a balance of contact interaction and excluded volume; Schellman JA; Changes in excluded volume and contact interaction with the surface of a protein have been suggested as mechanisms for the changes in stability induced by cosolvents . The aim of the present paper is to present an analysis that combines both effects in a quantitative manner . The result is that both processes are present in both stabilizing and destabilizing interactions and neither can be ignored . Excluded volume was estimated using accessible surface area calculations of the kind introduced by Lee and Richards . The change in excluded volume on unfolding, deltaX, is quite large . For example, deltaX for ribonuclease is 6.7 L in urea and approximately 16 L in sucrose . The latter number is greater than the molar volume of the protein . Direct interaction with the protein is represented as the solvent exchange mechanism, which differs from ordinary association theory because of the weakness of the interaction and the high concentrations of cosolvents . The balance between the two effects and their contribution to overall stability are most simply presented as bar diagrams as in Fig . 3 . Our finding for five proteins is that excluded volume contributes to the stabilization of the native structure and that contact interaction contributes to destabilization . This is true for five proteins and four cosolvents including both denaturants and osmolytes . Whether a substance stabilizes a protein or destabilizes it depends on the relative size of these two contributions . The constant for the cosolvent contact with the protein is remarkably uniform for four of the proteins, indicating a similarity of groups exposed during unfolding . One protein, staphylococcus nuclease, is anomalous in almost all respects . In general, the strength of the interaction with guanidinium is about twice that of urea, which is about twice that of trimethylamine-N-oxide and sucrose . Arguments are presented for the use of volume fractions in equilibrium equations and the ignoring of activity coefficients of the cosolvent . It is shown in the Appendix that both the excluded volume and the direct interaction can be extracted in a unified way from the McMillan-Mayer formula for the second virial coefficient.

Br J Dermatol, 2003 Jun, 148(6), 1222 - 9
Protein A immunoadsorption: a novel and effective adjuvant treatment of severe pemphigus; Schmidt E et al.; BACKGROUND: Pemphigus foliaceus (PF) and pemphigus vulgaris (PV) are autoimmune blistering skin diseases usually treated with high-dose systemic corticosteroids and other immunosuppressants that may cause severe side-effects . Plasmapheresis also has been demonstrated to be of benefit in the treatment of pemphigus . In contrast to plasmapheresis, staphylococcal protein A immunoadsorption (PA-IA) specifically removes immunoglobulin from the circulation, allows treatment of larger plasma volumes, and does not require the substitution of plasma components . OBJECTIVES: To determine the effectiveness and side-effects of PA-IA in patients with severe pemphigus . METHODS: Five patients with severe pemphigus (PV, n = 4; PF, n = 1) were treated by PA-IA . Three of these patients had been refractory to various treatment regimens . In addition to PA-IA, methylprednisolone, 0.5 mg x kg-1 body weight day-1 was given initially and subsequently tapered . RESULTS: In all patients, a dramatic clinical improvement was seen within 2 weeks after initiation of therapy . Patients were free of lesions after 3, 4, 4, 10 and 21 weeks of treatment, respectively . Concurrently, autoantibody levels decreased rapidly . CONCLUSIONS: PA-IA is a rational, effective, and safe adjuvant therapy for severe pemphigus and warrants wider use for this indication . A controlled study should compare side-effects and effectiveness of PA-IA with other treatment options for pemphigus.

Scand J Immunol, 2003 Jul, 58(1), 89 - 98
Aberrant T-cell receptor signalling of interferon-gamma- and tumour necrosis factor-alpha-producing cytotoxic CD8+ Vdelta1/Vbeta16 T cells in a patient with chronic neutropenia; Bank I et al.; We previously found that the peripheral blood (PB) mononuclear cells (MCs) (PBMCs) of a patient with chronic neutropenia contained an expanded population of cytotoxic CD8+ T cells using a variable (V) region delta1 gene product in the T-cell receptor-alpha (TCR-alpha) polypeptide {Vdelta1-constant(C)alpha+ T cells} . Sequencing of polymerase chain reaction (PCR) amplification products have now revealed a productive Vdelta1/joining (J)alphaIGRJa03/Calpha rearrangement of the TCR-alpha gene, predominantly associated with a Vbeta16/Dbeta2.1/Jbeta2.1/Cbeta2 TCR-beta gene, in these cells . Furthermore, we detected a markedly deficient proliferative response of the patient PBMCs to triggering with monoclonal antibodies (MoAbs) to the CD3 molecule, contrasting with a substantial response to the Vbeta3, 12, 14, 15, 17 and 20-specific staphylococcal enterotoxin B (SEB) superantigen, suggesting defective TCR-mediated activation of the Vdelta1+/Vbeta16+ clone . Moreover, whereas triggering of Vdelta1- T cells cultured with interleukin-2 (IL-2) by MoAb to the CD3 molecule enhanced proliferation, Vdelta1-Calpha+ T cells were inhibited by MoAbs to either CD3 or Vdelta1 . Vdelta1-Calpha+ T-cell clones spontaneously secrete interferon-gamma (IFN-gamma) and were further induced to release tumour necrosis factor (TNF-alpha) when triggered by anti-CD3 plus phorbol ester . Aberrant signalling by the clonotypic TCR together with the functional properties of the CD8+ Vdelta1+/Vbeta16+ clone may thus contribute to the immunohaematological abnormalities observed in this patient.

Cell Immunol, 2003 Apr, 222(2), 144 - 55
Staphylococcal enterotoxin B induces anergy to conventional peptide in memory T cells; Watson AR et al.; Microbial superantigens can alter host immunity through aberrant activation and subsequent anergy of responding naive T cells . We show here that the superantigen, staphylococcal enterotoxin B (SEB), directly induces tolerance in memory CD4 T cells . Murine naive and memory CD4(+) T cells were labeled with the fluorescent dye CFSE and the cells were exposed to SEB before they were cultured with specific peptide antigen . Memory, but not naive, T cells became anergic and did not respond to their cognate peptide antigen . The extent and duration of T cell receptor (TCR) clustering was similar to promote naive T cell activation and memory T cell anergy, suggesting similar TCR-SEB interactions led to distinct intracellular signaling processes in the two cell types . Like SEB, soluble anti-CD3 mAb does not stimulate memory cell proliferation . However, unlike SEB, soluble anti-CD3 mAbs did not induce anergy to cognate peptide . Anergy was directly visualized in vivo . CD4(+) memory T cells were identified in mice that had been administered SEB . The cells failed to proliferate in response to subsequent immunization with their cognate recall antigen . Hence, one mode of pathogen survival is the modulation of host immunity through selective elimination of memory T cell responses.

Protein Sci, 2003 Jul, 12(7), 1530 - 7
Effects of denaturants and substitutions of hydrophobic residues on backbone dynamics of denatured staphylococcal nuclease; Ohnishi S et al.; Analysis of residual dipolar couplings (RDCs) in the Delta131Delta fragment of staphylococcal nuclease has demonstrated that its ensemble-averaged structure is resistant to perturbations such as high concentrations of urea, low pH, and substitution of hydrophobic residues, suggesting that its residual structure is encoded by local side-chain/backbone interactions . In the present study, the effects of these same perturbations on the backbone dynamics of Delta131Delta were examined through (1)H-(15)N relaxation methods . Unlike the global structure reported by RDCs, the transverse relaxation rates R(2) were quite sensitive to denaturing conditions . At pH 5.2, Delta131Delta exhibits an uneven R(2) profile with several characteristic peaks involving hydrophobic chain segments . Protonation of carboxyl side chains by lowering the pH reduces the values of R(2) along the entire chain, yet these characteristic peaks remain . In contrast, high concentrations of urea or the substitution of 10 hydrophobic residues eliminates these peaks and reduces the R(2) values by a greater amount . The combination of low pH and high urea leads to further decreases in R(2) . These denaturant-induced increases in backbone mobility are also reflected in decreases in (15)N NOEs and in relaxation interference parameters, with the former reporting an increase in fast motions and the latter a decrease in slow motions . Comparison between the changes in chain dynamics and the corresponding changes in Stokes radius and the patterns of RDCs suggests that regional variations in backbone dynamics in denatured nuclease arise primarily from local contacts between hydrophobic side chains and local interactions involving charged carboxyl groups.

Transfusion, 2003 Jul, 43(7), 974 - 8
Monitoring of apheresis platelet bacterial contamination with an automated liquid culture system: a university experience; Brecher ME et al.; BACKGROUND: With 4 million platelet units transfused per year in the United States and with the current estimate of bacteria contamination rate in PLT units, it would be expected that 2000 to 4000 bacterially contaminated units are transfused and associated with 333 to 1000 cases of clinical sepsis . STUDY DESIGN AND METHODS: Apheresis platelets were sampled on Day 2 of storage (collection day=Day 0) and issue (or following outdate, Days 6-8) using a sterile connection device (SCD) to attach a sampling bag . Using aseptic technique and a laminar flow hood, bottles were inoculated and placed onto an automated liquid culture system (BacT/ALERT 3D Microbial Detection System) for 7 days . RESULTS: A total of 2397 apheresis PLT units were sampled . A triple apheresis collection was reactive within 14 hours of the Day 2 sampling (aerobic bottles) and the bags were removed from inventory . Staphylococcus epidermidis was identified in all three contaminated bags . Two double-apheresis collections were found to be contaminated with Proprionibacterium sp . after 6 days of incubation but had been transfused to four patients without discernible clinical sequelae . There was one false-positive aerobic bottle and one false-positive anaerobic result due to inadvertent contamination of a bottle . Thus, the overall true-positive rate was 7 of 2397 apheresis units (0.29%) with a true-positive rate for aerobic organisms of 0.13% and an anaerobic true-positive rate of 0.17% . The false-positive rate was 2 out of 4794 samplings (0.04%) or 2 out of 9588 bottles (0.02%) . CONCLUSION: This preliminary data suggests that the use of a SCD, aseptic technique, and a laminar flow hood is associated with a low rate of contamination . In no case did an issue (or outdate) detect contamination that was not detected by the Day 2 culture . Additional surveillance is necessary before we can conclude that a Day 2 sterile culture is truly predictive of an issue (or outdate) sterile culture . Bacterial culture surveillance of PLTs would be expected to save lives and may facilitate an extension in PLT storage.

Clin Exp Immunol, 2003 Jul, 133(1), 108 - 14
Modulation of cytokine release from colonic explants by bacterial antigens in inflammatory bowel disease; Dionne S et al.; The intestinal flora play an important role in experimental colitis and inflammatory bowel disease (IBD) . Using colonic explant cultures from 132 IBD and control subjects, we examined tumour necrosis factor-alpha (TNF-alpha), interleukin (IL)-1 and interleukin-1 receptor antagonist (IL-1RA) production in vitro in response to bacterial activators . Unstimulated TNF-alpha release was increased significantly in rectal biopsies from involved IBD tissue, correlating with inflammation severity . Whereas lipopolysaccharide (LPS) only moderately stimulated TNF-alpha production from inflamed tissue, pokeweed mitogen (PWM) induced its release in all groups, with a stronger response in involved IBD tissue . Superantigen staphylococcal enterotoxin A (SEA) had a similar, but weaker effect . SEB was observed to be the strongest inducer of TNF-alpha for all groups, again with a more marked response in inflamed tissue . Stimulated release of IL-1 was considerably less than for TNF-alpha . The superantigens' superior potency over LPS was not as marked for IL-1 as it was for TNF-alpha . In addition to IL-1, IL-1RA release was also triggered by the bacterial products . The net effect of activation on the IL-1RA/IL-1 ratio was relatively modest . Release of the proinflammatory cytokines TNF-alpha and IL-1, as well as that of the anti-inflammatory cytokine IL-1RA was increased by incubation of colonic tissue with bacterial factors . TNF-alpha production and release was increased significantly in involved colonic explants from IBD . SEB was even capable of inducing TNF-alpha release from uninvolved colonic tissue.

AIDS, 2003 May 23, 17(8), 1139 - 44
Quantitative ex vivo analysis of functional virus-specific CD8 T lymphocytes in the blood and genital tract of HIV-infected women; Kaul R et al.; BACKGROUND: CD8 T lymphocytes are important in HIV-1 control and mediate virus-specific immunity in the blood and genital tract . The induction and monitoring of mucosal CD8 cell responses will be an important component of HIV-1 vaccine trials, but information regarding the frequency, phenotype and function of genital tract CD8 cell responses is lacking . METHODS: Simultaneous blood and cervical cytobrush samples were obtained from 16 HIV-1-infected Kenyan sex workers . Epitope-specific CD8 T lymphocyte frequencies in the blood and genital tract were analysed after short-term peptide incubation and intracellular cytokine staining for interferon-gamma (IFN gamma) . RESULTS: Cervical sampling resulted in adequate cell numbers for analysis in 10/16 women . Background IFN gamma production was higher in CD3+/CD8+ lymphocytes from the genital tract than from blood (0.48% versus 0.1%; P < 0.01) . Responses to staphylococcal enterotoxin B were detected in cervical CD8 lymphocytes from 10/10 women, at a similar frequency to blood (16.7% in cervix and 13.3% in blood; P = 0.4) . HIV-1-specific responses were detected the cervix of 8/10 women, with a trend to higher response frequencies in the genital tract than blood (2.1% versus 0.8%; P = 0.09) . Co-expression of integrin CD103 (alpha E beta 7), a mucosal marker, was used to confirm the mucosal origin of cervical responses . CONCLUSIONS: Cytobrush sampling and intracellular cytokine staining is well suited to the analysis of cervical CD8 cell responses . The frequency of functional virus-specific CD3+/CD8+ T cells is similar in the genital tract and blood of HIV-1-infected women . The role of genital tract CD8 cell responses in HIV-1 control warrants further investigation.

Mol Endocrinol, 2003 Sep, 17(9), 1805 - 14 Epub 2003 Jun 20.
Tudor and nuclease-like domains containing protein p100 function as coactivators for signal transducer and activator of transcription 5; Paukku K et al.; Signal transducer and activator of transcription 5 (Stat5) plays a critical role in prolactin (PRL)-induced transcription of several milk protein genes . Stat5-mediated gene regulation is modulated by cooperation of Stat5 with cell type- and promoter-specific transcription factors as well as by interaction with transcriptional coregulators . Recently, the expression of a tudor and staphylococcal nuclease-like domains containing protein p100 was found to be increased in mammary epithelial cells during lactation in response to lactogenic hormones . p100 was initially identified as a transcriptional coactivator of the Epstein-Barr virus nuclear antigen 2 . In this study we investigated the potential role of p100 in PRL-induced Stat5-mediated transcriptional activation . PRL stimulation increased the p100 protein levels in HC11 mouse mammary epithelial cells . p100 did not affect the early activation events of Stat5, but p100 enhanced the Stat5-dependent transcriptional activation in HC11 cells . p100 associated with Stat5 both in vivo and in vitro, and the interaction was mediated by both the tudor and staphylococcal nuclease-like domains of p100 . Together these results suggest that p100 functions as a transcriptional coactivator for Stat5-dependent gene regulation and the existence of a positive regulatory loop in PRL-induced transcription, in which PRL stabilizes p100 protein, which in turn can cooperate with Stat5 in transcriptional activation.

Infect Immun, 2003 Jul, 71(7), 3724 - 9
Control of the oxidative burst of human neutrophils by staphylococcal leukotoxins; Colin DA et al.; The ability of staphylococcal two-component leukotoxins to induce an oxidative burst and/or to prime human polymorphonuclear cells (PMNs) was studied by using spectrofluorometry or flow cytometry . At sublytic concentrations, the HlgA-HlgB, HlgA-LukF-PV, LukS-PV-LukF-PV, and HlgC-LukF-PV combinations of leukotoxins, but not the LukS-PV-HlgB and HlgC-HlgB combinations, were able to induce H(2)O(2) production similar to the H(2)O(2) production induced by 1 micro M N-formyl-Met-Leu-Phe (fMLP) . In addition, when added at sublytic concentrations, all of the leukotoxin combinations primed PMNs for H(2)O(2) production induced by fMLP . Leukotoxin activation was dependent on the presence of Ca(2+) and was inhibited by wortmannin, an inhibitor of phosphatidylinositol 3-kinase, but not by N-methyl-L-arginine, an inhibitor of NO generation, which eliminates the possibility that NO plays a role in the action of leukotoxins . At higher concentrations, all leukotoxins inhibited H(2)O(2) production by PMNs activated by fMLP, phorbol 12-myristate 13-acetate (PMA), or the leukotoxins themselves . This inhibition was not related to the pore formation induced by leukotoxins . Intracellular release of H(2)O(2) induced by fMLP and PMA was not primed by leukotoxins but was inhibited . It seems that leukotoxin inhibition of H(2)O(2) release is independent of pore formation but secondary to an intracellular event, as yet unknown, triggered by leukotoxins.

J Gen Virol, 2003 Jul, 84(Pt 7), 1701 - 10
Antibody-mediated growth of infectious salmon anaemia virus in macrophage-like fish cell lines; Joseph T et al.; Infectious salmon anaemia virus (ISAV), a pathogen in marine aquaculture, belongs to the genus Isavirus, family Orthomyxoviridae . There is limited information on how ISAV interacts with host defences . To study ISAV-antibody interactions, virus neutralization (VN) assays were performed in the cell lines CHSE-214, SHK-1 and TO using three strains of ISAV and rabbit or fish anti-ISAV sera . Homologous VN titres of >1 : 1280 in CHSE-214 cells corresponded to titres of only 1 : 80 in the macrophage-like fish cell lines SHK-1 and TO, despite using 1000 and 2000 times less virus, respectively . However, rabbit antiserum to infectious pancreatic necrosis virus (IPNV) had a VN titre of 1 : 10,260 against IPNV in both CHSE-214 and TO cells . Poor ISAV neutralization in TO cells was attributed to Fc receptors mediating virus infectivity, because (1) neutralization by rabbit antiserum to ISAV was increased 48-fold in the presence of staphylococcal Protein A and (2) when using FITC-labelled virus and spectrofluorometry, a significant increase (P=0.018) in the intensity of fluorescence of intracellular virus was observed in assays of virus-antiserum mixtures in the absence of Protein A as compared to those in the presence of Protein A . Neutralization of ISAV with fish antisera was observed only in CHSE-214 cells, as Protein A could not restore neutralization in TO cells . These findings demonstrate for the first time antibody-mediated infection of macrophage-like fish cell lines by a fish virus, ISAV, and, as ISAV in Atlantic salmon targets leukocytic and endothelial cells, this may have implications for ISA pathogenesis and vaccination.

Int Immunol, 2003 Jul, 15(7), 885 - 92
Effector T cells have a lower ligand affinity threshold for activation than naive T cells; Kimachi K et al.; It has been previously established that effector and memory T cells are more sensitive to antigen stimulation than naive T cells . In this study, we compared the effect of ligand affinity on the activation of naive and effector T cells derived from pigeon cytochrome c (PCC)-specific TCR transgenic mice by stimulating these cells with a variety of ligands with widely differing antigenicity . The data obtained indicated the following . (i) The differences in antigen dose requirements for activation of naive and effector cells widened as the affinity of the antigen decreased . Most dramatically, peptides that were TCR antagonists for naive T cells were recognized as agonists by effector T cells . (ii) While both naive and effector T cells were activated by the bacterial superantigen staphylococcal enterotoxin A, specific for the transgenic TCR V(beta)3 chain, effector, but not naive, T cells were stimulated to proliferate by toxic shock syndrome toxin-1, a superantigen not previously described to be stimulatory for V(beta)3 T cells . (iii) Effector T cells, but not naive cells, proliferated in response to endogenous self-peptides presented by antigen-presenting cells in a syngeneic mixed lymphocyte reaction . Taken together these data indicate that effector T cells have a lower affinity threshold for activation than naive T cells . Further studies demonstrated that the heightened reactivity of effector T cells to low-affinity ligands declined progressively with repeated stimulations by antigen such that after repeated stimulation effector T cells were no longer stimulated by low-affinity ligands but recognized them as TCR antagonists similar to naive T cells.

Int Immunol, 2003 Jul, 15(7), 817 - 26
CD154-CD40-independent up-regulation of B7-2 on splenic antigen-presenting cells and efficient T cell priming by staphylococcal enterotoxin A; Eshima K et al.; It has been demonstrated that in vivo T cell priming requires CD154-CD40 interaction, which is suggested to be critical in the induction of co-stimulatory activities on antigen-presenting cells (APC) . In the current study, we demonstrate that in vivo administration of a high dose of a superantigen, staphylococcal enterotoxin A (SEA), could up-regulate B7-2 on most splenic APC independently of the CD154-CD40 interaction, followed by efficient expansion of SEA-reactive V(beta)3(+) T cells in CD154- or CD40-deficient mice . However, the CD154-CD40 interaction may be involved in SEA-mediated T cell activation, since a contribution of the CD154-CD40 interaction was observed when a lower dose of SEA was injected . CD154-independent T cell priming by SEA appeared also independent of the TRANCE-RANK pathway, which was shown to be capable of mediating CD154-independent activation of naive T cells during the infection of some viruses . These results indicate that SEA, which provokes rapid and efficient T cell responses without adjuvant, could utilize multiple CD154/TRANCE-independent pathways, to prime T cells.

Rev Panam Salud Publica, 2003 Apr, 13(4), 214 - 21
{Resistance to antimicrobials and combination therapy assessment in neonatal sepsis}; Espino Hernandez M et al.; OBJECTIVES: To estimate antibiotic resistance among bacterial strains isolated from patients with neonatal sepsis from January 1994 through December 1998, and to assess the predictive value of the checkerboard method for selecting treatment with combination antibiotic therapy in seriously ill patients . METHODS: The study of strain susceptibility was carried out by microdilution in broth, and the checkerboard method in broth trays was used to assess the efficacy of antibiotic combination therapy . RESULTS: Fifty per cent of the strains of coagulase-negative Staphylococcus spp . and 37% of S . aureus strains, which are the main pathogens involved in neonatal sepsis, were resistant to methycillin . Of these strains, 94.5% were resistant to several antibiotics . Four strains of coagulase-negative Staphylococcus spp . and one strain of Enterococcus faecium were resistant to vancomycin (minimal inhibitory concentration = 128 micro g/mL) . We noted a high probability of clinical efficacy (P < 0.01) with antibiotic combinations having a synergistic effect in vitro, whereas antagonic reactions were seen in close association with a highly significant probability of clinical therapeutic failure . DISCUSSION: The main cause of neonatal sepsis in the Neonatal Intensive Care Unit at "America Arias" Hospital in Havana, Cuba, were strains of Staphylococcus resistant to methycillin and other antibiotics . In seriously ill patients (according to policy established at this institution), the first line of treatment is a combination of an aminoglycoside antibiotic and a beta-lactamic antibiotic . This study showed that the clinical efficacy of combination antibiotic therapy is not determined by the resistance pattern of the strain and that good clinical results can be attained, in all likelihood, when a synergistic effect can be shown in vitro.

Hum Gene Ther, 2003 May 20, 14(8), 789 - 802
Antibody-mediated targeting of replication-competent retroviral vectors; Tai CK et al.; Replication-competent murine leukemia virus (MLV) vectors can be engineered to achieve high efficiency gene transfer to solid tumors in vivo and tumor-restricted replication, however their safety can be further enhanced by redirecting tropism of the virus envelope . We have therefore tested the targeting capability and replicative stability of ecotropic and amphotropic replication-competent retrovirus (RCR) vectors containing two tandem repeats from the immunoglobulin G-binding domain of Staphylococcal protein A inserted into the proline-rich "hinge" region of the envelope, which enables modular use of antibodies of various specificities for vector targeting . The modified envelopes were efficiently expressed and incorporated into virions, were capable of capturing monoclonal anti-HER2 antibodies, and mediated efficient binding of the virus-antibody complex to HER2-positive target cells . While infectivity was markedly reduced by pseudotyping with targeted envelopes alone, coexpression of wild-type envelope rescued efficient cellular entry . Both ecotropic and amphotropic RCR vector/anti-HER2 antibody complexes achieved significant enhancement of transduction on murine target cells overexpressing HER2, which could be competed by preincubation with excess free antibodies . Interestingly, HER2-expressing human breast cancer cells did not show enhancement of transduction despite efficient antibody-mediated cell surface binding, suggesting that target cell-specific parameters markedly affect the efficiency of post-binding entry processes . Serial replication of targeted vectors resulted in selection of Z domain deletion variants, but reduction of the overall size of the vector genome enhanced its stability . Application of antibody-mediated targeting to the initial localization of replication-competent virus vectors to tumor sites will thus require optimized target selection and vector design.

J Endotoxin Res, 2003, 9(2), 119 - 23
Macrophage migration inhibitory factor (MIF) regulates host responses to endotoxin through modulation of Toll-like receptor 4 (TLR4); Roger T et al.; The cytokine macrophage migration inhibitory factor (MIF) has emerged recently as an important mediator of inflammation and innate immunity . MIF is rapidly released by macrophages after stimulation with microbial products and pro-inflammatory cytokines and, in turn, stimulates the production of pro-inflammatory mediators by immune cells . Immunoneutralization of MIF or deletion of the Mif gene was shown to protect animals from lethal endotoxemia, staphylococcal toxic shock and septic shock in experimental models of bacterial peritonitis . To investigate the function of MIF in innate immunity, we studied the response of macrophages expressing reduced levels of MIF to microbial products . These cells were generated by transduction of an antisense MIF adenovirus or by stable transfection with an antisense MIF plasmid or were obtained from MIF-knockout mice . MIF-deficient macrophages were shown to be hyporesponsive to stimulation with LPS and Gram-negative bacteria . The defect was associated with a down-regulation of Toll-like receptor 4 (TLR4), the signal transducing molecule of the LPS receptor complex . Immunoneutralization of extracellular MIF decreased TLR4 expression and responses of macrophages to LPS, indicating that MIF may exert autocrine effects . These findings identify an important role for MIF in innate immunity and provide a rationale for the development of anti-MIF strategy for the treatment of patients with Gram-negative septic shock.

Pediatr Infect Dis J, 2003 Jun, 22(6), 545 - 52
Optimizing blood culture practices in pediatric immunocompromised patients: evaluation of media types and blood culture volume; Gaur AH et al.; BACKGROUND: The optimal use of blood cultures to determine the etiology of febrile episodes in neutropenic children has not been well-defined . METHODS: Single volume blood cultures using the Pediatric ISOLATOR System (ISO), were compared with variable, weight-based culture volumes using the BACTEC 9240 Culture System (BAC) . Additionally the value of routinely inoculating the BACTEC MYCO/F LYTIC culture vial (MFL) as well as the BACTEC AEROBIC/F culture vial (AF) was examined . RESULTS: A total of 2620 cultures had both ISO and BAC inoculated; 182 cultures were positive (7.0% of cultures); 97.8% of positive cultures were detected by the BAC (AF and/or MFL) vs.46.2% detected by the ISO . The advantage of the BAC over the ISO was statistically significant for overall recovery of isolates and bloodstream infections, including most individual organism categories . There were only two instances (one each of histoplasmosis and candidemia) in which a blood stream infection was detected by ISO only . All the isolates judged to be contaminants were recovered by BAC only . AF detected significantly more coagulase-negative Staphylococcus spp . than the MFL . Of the isolates 16%, representing 14% of the bloodstream infections (including Gram-negative infections), were detected by the MFL only . Infections were detected more quickly by BAC than by ISO (P < 0.0001) . Among the BAC media types, AF was faster than MFL (P < 0.0001) . CONCLUSIONS: Optimal yield of blood cultures in immunocompromised pediatric patients included the use of BAC with a weight-based, graduated volume of culture inoculation and routine use of both AF and MFL.

Nucleic Acids Res, 2003 Jun 15, 31(12), 3027 - 32
A DNA Spiegelmer to staphylococcal enterotoxin B; Purschke WG et al.; Bacterial staphylococcal enterotoxin B is involved in several severe disease patterns and it was therefore used as a target for the generation of biologically stable mirror-image oligonucleotide ligands, so called Spiegelmers . The toxin is a 28 kDa protein consisting of 239 amino acids . Since the full-length protein is not accessible to chemical peptide synthesis, a stable domain of 25 amino acids was identified as a suitable selection target . DNA in vitro selection experiments were carried out against the equivalent mirror-image D-peptide domain resulting in high affinity D-DNA aptamers . As expected, the corresponding enantiomeric L-DNA Spiegelmer showed comparable binding characteristics to the L-peptide domain . Moreover, the Spiegelmer bound the whole protein target with only slightly reduced affinity . Dissociation constants of both peptide-oligonucleotide complexes were measured in the range of 200 nM, whereas the Spiegelmer binding to the full-length protein was determined at approximately 420 nM . These data demonstrate the possibility to identify Spiegelmers against large protein targets by a domain approach.

Vaccine, 2003 Jun 20, 21(21-22), 2791 - 6
Correlation of body temperature with protection against staphylococcal enterotoxin B exposure and use in determining vaccine dose-schedule; Boles JW et al.; The immunoprotective potential of a recombinant vaccine against the incapacitating effect of aerosolized staphylococcal enterotoxin B (SEB) in nonhuman primates is reported . SEB belongs to a family of structurally related superantigens responsible for serious, life threatening pathologies . Injecting the recombinant SEB vaccine did not induce temperature elevation in rhesus monkeys, a classical symptom of toxic-shock syndrome . No temperature elevation was noted following injection with control tetanus toxoid . In addition to 100% survival, we observed a clear correlation between vaccine dose and mitigation of temperature elevation after a lethal SEB aerosol challenge . We conclude that the recombinant SEB vaccine is non-pyrogenic and that monitoring changes in body temperature is an important biomarker of toxic shock in a primate animal model.

Pediatr Crit Care Med, 2001 Jan, 2(1), 57 - 62
Complications of central venous catheterization in critically ill children; Casado-Flores J et al.; OBJECTIVE: Analysis of central venous catheterization complications in different access sites with the Seldinger technique . Patients and Methods: A prospective study (May 1992 through December 1996) of 308 central venous catheterizations in children of different ages in a pediatric intensive care unit . RESULTS: Access sites were the subclavian vein (76.3%), femoral vein (20.4%), and jugular vein (3.2%) . The frequency of catheter placement-related complications was 22%, and the frequency of serious catheter placement-related complications was 2.9% (pneumothorax 1.9%, hemothorax 1%) . Catheter placement-related complications were more common in the subclavian than in the femoral vein (chi-square, p =.02) for the larger bore catheters (chi-square, p =.01) and for the higher number of attempts (Student's t -test, p <.001) . Catheter placement-related complications were not related to the age, weight, or whether the procedure was performed by the staff physician or resident . The overall complication rate for maintenance-related complications was 36% . Maintenance-related complications were more common in younger children (Student's t -test, p =.03) . The most frequent maintenance-related complications were mechanical complications (catheter obstruction and central venous thrombosis), and these were higher for femoral access (chi-square, p <.01) and for catheters indwelling for a longer period of time . Infection was found in 5.8% of patients, mainly due to Staphylococcus epidermidis . Infection was not related to the site of venous access or to the length of time the catheter was left indwelling . CONCLUSIONS: Central venous catheterization can be performed readily in children of all ages with an acceptable degree of risk . The immediate complications were more frequent and severe for subclavian vein catheterization, and the highest risk factor was the number of attempts at catheter insertion . Although the most frequent late complications were mechanical, which were higher for the femoral access and long-indwelling catheters, femoral catheters can be left indwelling for longer periods if routine ultrasound follow-up is performed . Infectious complications were independent of the venous access site or the duration of catheterization.

Rev Med Chil, 2003 Mar, 131(3), 237 - 50
{Infectious endocarditis: morbi-mortality in Chile . Results of the National Cooperative Study of Infective Endocarditis (1998-2002)}; Oyonarte M et al.; BACKGROUND: Infective endocarditis is a severe condition, with a mortality that fluctuates between 16 and 25% in the Metropolitan area of Chile . AIM: To perform a prospective assessment of clinical and microbiological features of patients with infective endocarditis in Chile . MATERIAL AND METHODS: Collaborative study of regional hospitals in the whole country and teaching hospitals in Santiago . Patients with a possible or definitive infective endocarditis, according to Duke's criteria, were included in the protocol and a structured data entry form was completed . RESULTS: Three hundred twenty one patients (65% male, mean age 49 +/- 16.5 years) were studied . According Duke's criteria, 89% had a definitive and 11% a possible endocarditis . The subacute form occurred in 64% of patients . The most frequent predisposing cardiopathies were rheumatic in 25%, prosthetic valves in 15% and congenital in 13% . There was no evidence of cardiopathy in 20% . Twenty percent of patients were on hemodialysis, 11% were diabetic and only one patient abused intravenous drugs . The most frequent complication was cardiac failure in 59% of cases, followed by renal failure in 32% and embolism in 28% . The most frequent causing organism was coagulase positive Staphylococcus in 35% . Blood cultures were negative in 28% of cases from the metropolitan region, in 56% of cases from the north and 38% of cases from the south . Echocardiographic diagnosis was done in 92% of cases . Aortic valve was involved in 42% and mitral valve in 29% . Successful antimicrobial treatment was achieved in 59% of patients . Thirty five percent of patients were subjected to surgical procedures with a 78% survival . Overall mortality was 29% . Univariate analysis identified sepsis, an age over 60 years and the presence of cardiac or renal failure as prognostic indicators of mortality . On multivariate analysis, the identified prognostic indicators were the presence of sepsis, renal failure, mitroaortic involvement associated to combined surgery and failure of antimicrobial treatment not associated to surgery . CONCLUSIONS: Subacute form is the most common presentation of infective endocarditis and rheumatic valve disease is the most common underlying cardiac lesion . Intravenous drugs users infective endocarditis is exceptional in Chile . The most frequent causing agent is coagulase positive Staphylococcus and the most frequent complication is cardiac failure . Surgical and overall mortality were 22 and 29% respectively . Sepsis, renal failure, combined surgical procedures, failure of medical treatment were identified as prognostic indicators of mortality.

Mol Cell Probes, 2003 Apr-Jun, 17(2-3), 125 - 6
A simple and rapid fluorescence-based immunoassay for the detection of staphylococcal enterotoxin B; Khan AS et al.; The bioterrorism threat is perceived to be a real challenge to our nation's security . This threat has necessitated the design of better and faster assays for the detection of biothreat agents including staphylococcal enterotoxin B (SEB), a causative agent of food poisoning . This study describes a simple, fast and highly sensitive fluorescence-based immunoassay, in which the antibody is fluorescently-labeled for use in this assay . Use of labeled antibodies resulted in very low level of detection of SEB, 100 pg/well . This method is four times faster than classical and conventional enzyme-linked immunosorbent assay (ELISA).

Blood Purif, 2003, 21(3), 225 - 31
A switch to high-flux helixone membranes reverses suppressed interferon-gamma production in patients on low-flux dialysis; Lonnemann G et al.; Long-term hemodialysis (HD) induces an inflammatory response and is associated with a suppressed cellular immune response manifested, in part, by impaired interferon (IFN-gamma) production . We investigated the effect of high-flux HD using the synthetic Helixone membrane and ultrafiltered dialysate on plasma levels of inflammatory mediators and on the whole blood production of IFN-gamma . METHODS: Twelve ESRD patients were dialyzed under low-flux HD (polysulfone F6) and again after 6 weeks of high-flux HD (Helixone FX100) . Ultrafiltered bicarbonate dialysate without bacterial growth and no detectable endotoxin was used throughout the study . Plasma levels of urea, albumin, beta(2)-microglobulin (beta(2)-m), interleukin (IL)-6, C-reactive protein (CRP), IL-1 receptor antagonist (IL-1Ra), IL-18, and IL-18-binding protein (IL-18BP) were measured . In addition, the Staphylococcus epidermidis-induced production of IFN-gamma and IL-18 was assessed in whole blood cultures of HD patients as well as in 9 healthy subjects . RESULTS: Plasma levels of urea, albumin, IL-6, IL-1Ra and CRP were not significantly different between high-flux and low-flux HD . In contrast, beta(2)-m levels decreased significantly by 31% with high-flux Helixone (p < 0.002) . Stimulated whole blood production of IFN-gamma was reduced in low-flux HD but increased to near normal levels after 6 weeks of high-flux HD . Plasma levels of free IL-18 and its specific inhibitor IL-18BP were not different between the two dialyzer membranes . CONCLUSION: Compared to low-flux polysulfone HD with ultrafiltered dialysate, high-flux HD with the synthetic Helixone membrane did not result in a significant change in plasma levels of proinflammatory (IL-6, CRP, IL-18) and anti-inflammatory (IL-1Ra, IL-18BP) cytokines . However, high-flux HD restores whole blood IFN-gamma production without significant changes in free IL-18 . Therefore, the immune modulation in high-flux HD is likely due to removal of inhibitors of IFN-gamma production other than IL-18BP .

Acta Physiol Scand, 2003 Jun, 178(2), 155 - 63
Strength, workload, anaerobic intensity and the immune response to resistance exercise in women; Miles MP et al.; AIM: The mechanism linking exercise intensity to the magnitude of the immune response is not completely understood . The purpose of this investigation was to determine whether the immune response to resistance exercise was associated with (1) changes in workload or (2) anaerobic exercise intensity . METHODS: Previously untrained women underwent 6 months of resistance training for lower and upper body (TOTAL, n = 34) or for upper body alone (UPPER, n = 30) . Lymphocyte subsets {T (CD3+), CD4+, CD8+, NK and B}, functional markers (CD45RA+ and CD45RO+), and mitogen (phytohemagglutinin-M, concanavalin A and pokeweed mitogen) and superantigen (staphylococcus a . cowans)-stimulated proliferation were measured from blood samples collected pre- and post-exercise for a squat resistance exercise consisting of six sets of 10 repetitions at 75% of one repetition maximum . This protocol was performed before (T0) and after 3 (T3) and 6 months (T6) of training . RESULTS: Lymphocyte recruitment to the circulation and proliferation following resistance exercise did not differ between training groups at any time, although the TOTAL group performed at a higher workload as training progressed . With respect to anaerobic intensity, exercise-induced increases in NK, CD4+, CD8+ and B lymphocyte concentrations were 42 (P = 0.07), 76 (P < 0.05), 72 (P < 0.05) and 242% (P < 0.01) greater in women in the highest compared with the lowest post-exercise lactate quartiles . Lymphocyte proliferation did not differ between lactate quartiles . CONCLUSIONS: Anaerobic intensity, rather than increased strength and workload, is associated with the number of lymphocytes recruited to the circulation, but not T and B cell proliferation responses.

Saudi Med J . 2003 May;24(5 Suppl):S35.
CT scan diagnosis of extralobar pulmonary sequestration: angiography not required; Wassia HL et al.; We present a 2-day-old newborn infant with extrapulmonary sequestration manifesting itself as persistent staphylococcus epidermidis pneumonia and high output cardiac failure . The diagnosis was established by a CT scan of the chest without angiography . Radiographic evaluation of pulmonary sequestration includes delineation of the aberrant systemic feeding artery and the venous drainage . Color Doppler ultrasound, spiral CT scan and MRI angiography are often sufficient.

Zhonghua Er Bi Yan Hou Ke Za Zhi, 2002 Aug, 37(4), 264 - 6
{Effect of staphylococcal enterotoxin B on ion electrophysiology and permeability in rabbit maxillary sinus epithelium}; Liu T et al.; OBJECTIVE: To study the barrier role of staphylococcal enterotoxin B (SEB) in rabbit maxillary sinus mucosa . METHODS: Rabbit maxillary sinus mucosal membrane was dissected under a surgical microscope . The short-circuit current (Isc), conductance (G) and its permeability to horseradish peroxidase (HRP) were evaluated by employing Ussing chamber technique . RESULTS: The stimulation of SEB enhanced Isc, G and its permeability to HRP of sinus epithelial barrier . The same results were obtained by applying tumor necrosis factor alpha (TNF-alpha) to rabbit maxillary sinus . Increase in TNF-alpha level in sinus mucosa was observed by the stimulation of SEB . Those pathophysiological abnormalities of sinus mucosa were abolished by pretreatment with anti-TNF-alpha antibody . CONCLUSION: The SEB has potential effects on rabbit maxillary sinus mucosal barrier, activates the immune cells in the mucosa, and plays certain roles in maxillary sinusitis.

FEMS Immunol Med Microbiol, 2003 Jun 10, 37(1), 45 - 51
Apotransferrin administration prevents growth of Staphylococcus epidermidis in serum of stem cell transplant patients by binding of free iron; von Bonsdorff L et al.; We investigated the effect of free, non-transferrin-bound iron occurring in haematological stem cell transplant patients on growth of Staphylococcus epidermidis in serum in vitro, and prevention of bacterial growth by exogenous apotransferrin . S . epidermidis did not grow in normal serum at inoculated bacterial densities up to 10(3) cfu ml(-1) but slow growth could be detected at higher initial inocula . Addition of free iron abolished the growth-inhibitory effect of serum, whereas addition of apotransferrin again restored it . Appearance of free iron and loss of growth inhibition coincided in patient serum samples taken daily during myeloablative therapy . Intravenously administered apotransferrin effectively bound free iron and restored the growth inhibition in patient sera . The results suggest that exogenous apotransferrin might protect stem cell transplant patients against infections by S . epidermidis and possibly other opportunistic pathogens.

Childs Nerv Syst, 2003 Sep, 19(9), 683 - 5 Epub 2003 May 27.
A case of neonatal cerebellar abscess; Tsutsumi S et al.; CASE REPORT: A 1-month-old boy presented with repeated vomiting without signs of infection and was eventually found to have staphylococcal cerebellar abscess . The initial diagnosis was hypertrophic pyloric stenosis . RESULT: . He was successfully treated by drainage surgery combined with intravenous antibiotic therapy . CONCLUSIONS: . Neonatal brain abscess, particularly cerebellar abscess, is seldom encountered, but may cause disability in later years even after successful treatment . Early diagnosis and appropriate treatment are mandatory for these patients.

Spine, 2003 May 15, 28(10), E194 - 7
Full-thickness thoracic laminar erosion after posterior spinal fusion associated with late-presenting infection; Tribus CB et al.; STUDY DESIGN: A case report describes unilateral complete laminar erosion of the caudal thoracic spine and late-presenting infection in a patient 10 years after anteroposterior reconstruction for scoliosis . OBJECTIVE: To present an unusual but significant complication that may occur after implantation of spinal instrumentation . SUMMARY OF BACKGROUND DATA: The reported patient presented with a deep infection and persistent back pain 10 years after successful anteroposterior reconstruction for adult idiopathic scoliosis . Delayed onset infections after implantation of spinal instrumentation are infrequent, yet when present, often require hardware removal . METHODS: The case of a 51-year-old woman who underwent irrigation and debridement for a late-presenting infection and removal of posterior hardware 10 years after her index procedure is presented . Interoperatively, it was noted that full-thickness laminar erosion was present from T4 to T12 . RESULTS: The patient was taken to the operating room for wound irrigation, debridement, and hardware removal . It was discovered that a Cotrel-Dubousset rod placed on the convexity of the curve had completely eroded through the lamina of T7-T12 . Infectious material was found along the entire length of both the convex and concave Cotrel-Dubousset rods . Intraoperative cultures grew Staphylococcus epidermidis and Propionibacterium acnes . Intravenous and oral antibiotics were administered, resulting in resolution of the infection and preoperative pain . CONCLUSIONS: The exact role of late-presenting infection with regard to the laminar erosion and rod migration seen in this case remains to be elucidated . However, the authors believe the primary cause of bony erosion was mechanical in origin . Regardless, most spine surgeons will treat many patients who have had posterior spinal implants and will perform hardware removal on a significant number of these patients during their careers . A full-thickness laminar erosion exposes the spinal cord to traumatic injury during hardware removal and debridement . This case is presented as a cautionary note to help surgeons become cognizant of a potentially devastating complication.

Medicina (Kaunas), 2003, 39 Suppl 1, 70 - 4
{Incidence of peritonitis and catheter exit site infection in children undergoing automatic continuous cyclic peritoneal dialysis}; Baciulis V et al.; The aim of this study was to find out the clinical aspects of peritonitis and catheter exit site infection in children undergoing continuous cyclic peritoneal dialysis . The incedence of peritonitis and catheter related infections were reviewed in 8 children on continuous cyclic peritoneal dialysis over a mean period of 14.9+/-15.8 months . Peritonitis occurred in 4 children . There were 14 episodes of peritonitis . The mean time from starting dialysis to the first episode of peritonitis was 1.9+/-1.0 months . The incidence of peritonitis was 1 episode in 9 treatment months . Gram-negative organisms (Enterococcus spp.) were responsible for the majority of episodes (35.7%) of peritonitis and gram-positive bacteria mainly caused the catheter exit site infections (Staphylococcus epidermidis) . The incidence of catheter exit site infection was one episode in 8 treatment months . Recurrent peritonitis was present in 1 case . Most patients with dialysis-related peritonitis and catheter exit site infection responded to antibiotic therapy . Catheter was replaced in 2 patients . The mortality rate was 2 out of 8 patients but none of the deaths were related to peritonitis . Patients with exit site infection had 7 times higher risk than those without developing peritonitis.

Protein Sci, 2003 Jun, 12(6), 1232 - 46
Automated protein fold determination using a minimal NMR constraint strategy; Zheng D et al.; Determination of precise and accurate protein structures by NMR generally requires weeks or even months to acquire and interpret all the necessary NMR data . However, even medium-accuracy fold information can often provide key clues about protein evolution and biochemical function(s) . In this article we describe a largely automatic strategy for rapid determination of medium-accuracy protein backbone structures . Our strategy derives from ideas originally introduced by other groups for determining medium-accuracy NMR structures of large proteins using deuterated, (13)C-, (15)N-enriched protein samples with selective protonation of side-chain methyl groups ((13)CH(3)) . Data collection includes acquiring NMR spectra for automatically determining assignments of backbone and side-chain (15)N, H(N) resonances, and side-chain (13)CH(3) methyl resonances . These assignments are determined automatically by the program AutoAssign using backbone triple resonance NMR data, together with Spin System Type Assignment Constraints (STACs) derived from side-chain triple-resonance experiments . The program AutoStructure then derives conformational constraints using these chemical shifts, amide (1)H/(2)H exchange, nuclear Overhauser effect spectroscopy (NOESY), and residual dipolar coupling data . The total time required for collecting such NMR data can potentially be as short as a few days . Here we demonstrate an integrated set of NMR software which can process these NMR spectra, carry out resonance assignments, interpret NOESY data, and generate medium-accuracy structures within a few days . The feasibility of this combined data collection and analysis strategy starting from raw NMR time domain data was illustrated by automatic analysis of a medium accuracy structure of the Z domain of Staphylococcal protein A.

Biochim Biophys Acta, 2003 May 13, 1627(1), 1 - 6
Glutamine is incorporated at the nonsense codons UAG and UAA in a suppressor-free Escherichia coli strain; Nilsson M et al.; Readthrough of the nonsense codons UAG, UAA, and UGA is seen in Escherichia coli strains lacking tRNA suppressors . Earlier results indicate that UGA is miscoded by tRNA(Trp) . It has also been shown that tRNA(Tyr) and tRNA(Gln) are involved in UAG and UAA decoding in several eukaryotic viruses as well as in yeast . Here we have investigated which amino acid(s) is inserted in response to the nonsense codons UAG and UAA in E . coli . To do this, the stop codon in question was introduced into the staphylococcal protein A gene . Protein A binds to IgG, which facilitates purification of the readthrough product . We have shown that the stop codons UAG and UAA direct insertion of glutamine, indicating that tRNA(Gln) can read the two codons . We have also confirmed that tryptophan is inserted in response to UGA, suggesting that it is read by tRNA(Trp).

Immunology, 2003 Jun, 109(2), 246 - 54
Soluble FcgammaRIIa inhibits rheumatoid factor binding to immune complexes; Wines BD et al.; Soluble low-affinity receptors for IgG are known to inhibit immune complex (IC)-mediated inflammation, and expression by leukocytes is elevated in several inflammatory diseases . Immunoglobulin M (IgM) rheumatoid factors (RF), anti-Fc autoantibodies, are found in autoimmune diseases, such as rheumatoid arthritis (RA), as well as in normal immune responses . This study demonstrated that soluble FcgammaRIIa inhibits the interaction of rheumatoid factors with ICs . The recombinant soluble low-affinity FcgammaR, rsFcgammaRIIa, partially inhibited (30-70%) the rate of precipitation of soluble ICs by RF-positive RA sera . This required the normal interaction of FcgammaRIIa with Fc as the effect could be abrogated with the Fab fragment of the blocking mAb IV-3 . Furthermore, rsFcgammaRIIa partially inhibited (40%) the binding of a monoclonal IgM RF (RF-AN) to an IC formed by IgG2 antibody binding to an antigen-coated biosensor chip . Since RF-AN has been characterized by crystallography to bind to the CH2/CH3 interface of the IgG-Fc, and leukocyte FcgammaRIIa binds to a distinct site centred on the lower hinge, this inhibition is uncompetitive . Some inhibition (15%) of staphylococcal protein A binding to IC was also observed . As soluble FcgammaRIIa disrupts Fc:Fc interactions in IgG-ICs, we propose that this alteration of the IC also reduces the accessibility of Fc portions in the IC, resulting in the partial inhibition of ligands, particularly IgM RF, which bind Fc . We propose that the high concentrations of soluble FcgammaR found during inflammation can affect the properties of ICs and their interaction with the immune system.

Arerugi, 2003 Apr, 52(4), 434 - 43
{Oral therapy for atopic dermatitis in year 2000 . Questionnaire Study to Boarding Members of Japanese Society for Allergology}; Aoki T et al.; Questionnaires were sent to boarding members of Japanese Society for Allergology (response rate 65.2%; 49 dermatologists {Derma} and 101 pediatricians {Pedia} ) to ask the order of choice in therapy attitudes, in the oral in comparison to the local therapy and in the oral therapy only . As for therapy attitudes, "suppress inflammation" (Infla), "reduce staphylococcal colonization" (Staph), "inhibit scratching" (Scrat), "guard skin from irritation" (Guard), "avoid allergen" (Avoid), and "support psychologically" (Psych) were presented . The order of choice was Guard, Infla, Scrat and Avoid for the first four in both Derma and Pedia . The last 2 were Psych and Staph in Derma and vice versa in Pedia . However, the lowest adoption rate was 76.2% in Derma and 85.8% in Pedia . This suggests that wide therapy attitudes were taken in mind in the therapy of atopic dermatitis in the both specialites . The adoption rate was significantly (P>0.05) higher in Pedia than Derma with Staph, Avoid, Scrat and Psych . As for the oral versus local therapy, "antihistamine" (A-hist), "antiallergy" (A-aller), "sedative" (Sedat), "hypnotic" (Hypno), "corticosteroid hormone" (O-steroid) and "antibacterial drug" (A-bact) were presented for the oral therapy, and "skin care medicine" (SkinC), "NSAID" (t-NSAID), "corticosteroid ointment" (S-oint) and "disinfectant" (Disinf) were for the topical therapy . The adoption rate was in the order of SkinC as well as S-oint followed by A-aller and A-hist both in Derma and Pediat . These choices were over 60% in Derma and over 75% in Pediat . Disinf and t-NSAID followed these with over 50% of adoption rate in Pediat, while all others were 30.6% or less in Derma . Thus, in Derma 4 therapies (2 local and 2 oral) were thought to be the first rank therapy but in Pediat additional 2 therapies (Disinf, t-NSAID) were comparable to the first rank therapy in Derma . As for the oral therapy only, A-aller and A-hist were adopted at high rates both in Derma and Pediat, but all other therapies that included 5 additional therapies; "kampo drug", "antimycotic drug", "vitamine", "arachidonic acid cascade controlling drug" and "immunosuppressant" were adopted in only low rates . Experience of oral corticosteroid therapy was about three quarters in Derma but a quarter in Pediat . Conditions for prescribing oral corticosteroid were mostly "extreme severity" and "acute generalized aggravation" both in Derma and Pediat . About 40% of doctors both in Derma and Pediat who ever prescribed oral corticosteroid were afraid that corticosteroid might not be stopped when starting the therapy . However, only short-term oral corticosteroid therapy is described in the guidelines of this society for the treatment of atopic dermatitis (1995) . It is hoped to standardized oral corticosteroid therapy for long-lasting extremely severe atopic dermatitis.

Vet Rec, 2003 May 3, 152(18), 558 - 62
Serum amyloid A in the serum and milk of ewes with mastitis induced experimentally with Staphylococcus epidermidis; Winter P et al.; Mastitis was induced experimentally in ewes with Staphylococcus epidermidis, and the concentrations of serum amyloid A (SAA) in milk and serum, and the somatic cell counts and bacteria in the milk were determined for up to 10 weeks in two experiments, each examining five infected and five control ewes . The somatic cell counts peaked eight hours after infection and preceded an increase in SAA in milk . A maximum concentration of 6460 microg/ml SAA was recorded in milk from the infected sheep, compared with a mean concentration of 1.4 microg/ml in the control sheep . The mean peak concentration of SAA in serum (206.8 microg/ml) occurred earlier (one day after infection) than in milk . The serum concentration of SAA in the healthy animals ranged from 0 to 29.4 microg/ml . There was no correlation between the concentrations of SAA in serum and milk.

J Pharmacol Toxicol Methods, 2002 Jul-Aug, 48(1), 31 - 9
A new screening model for safety evaluation of superantigen-antibody recombinant fusion proteins (mAb Fab-SEA/E) using telemetric monitoring in conscious rabbits; Ilback NG et al.; INTRODUCTION: Carcinoma recognising monoclonal antibodies (mAb) and mutated forms of the T-cell-activating bacterial staphylococcal enterotoxin A/E (SEA/E) have been combined in single hybrid constructs (mAb Fab-SEA/E) . By introducing substitutions in an MHC class II binding site, these harmful toxins can be converted into tolerable immunotoxins . Rabbits and humans are sensitive to SE toxins, and cardiovascular effects in rabbits are similar to those seen in septic shock in man . A new screening model using telemetry in conscious rabbits was applied in the safety evaluation of different mAb Fab-SEA/E constructs administered intravenously . METHODS: Telemetry transmitters were implanted in the peritoneal cavity of animals with a pressure catheter in the aorta and electrodes for ECG recording subcutaneously following administration of mAb Fab-SEA/E constructs intravenously . RESULTS: The responses in body temperature, heart rate, and blood pressure varied depending on the treatment regimen and the mutations of the drug given . For example, 25 micro g/kg of C215 Fab-SEAmut9 were given as a first treatment cycle on days 1, 5, and 7 and as a second treatment cycle on days 13-15 . The first dose induced high fever, whereas the second and third doses induced fever responses more rapidly and were of lower and shorter duration . The second treatment cycle, starting on day 13, did not induce any responses probably due to anti-SEA antibodies formed because of the treatment . Another construct, 5T4 Fab-SEA/E-11 at 50 micro g/kg, induced a similar response as C215 Fab-SEAmut9 on days 1, 5, and 7 . In this case, the pharmacologic response was still present on days 13-15, though no clinical signs developed or no formation of anti-SEA antibodies occurred . When 50 micro g/kg of 5T4 Fab-SEA/E-11 was administered once daily for 4 days, body temperature after the first dose increased slowly during the first 24 h, whereas the second to fourth doses induced more rapid and higher responses . The fourth dose of another compound, K305 Fab-SEA/E-11 (50 micro g/kg), induced an even more pronounced response both in magnitude and in duration as well as in adverse clinical signs . DISCUSSION: By using continuous telemetric registration in the rabbit as a tool in superantigen-antibody (mAb Fab-SEA/E) drug selection, it has been possible to evaluate the dynamics of drug-induced immune effects (fever) and concomitant engagement of the cardiovascular system, conditions that are essential before clinical trials can be initiated.

Chin J Traumatol, 2003 Jun, 6(3), 167 - 70
Traumatic endophthalmitis following penetrating ocular injuries with retained intraocular foreign bodies; Jiang CH et al.; OBJECTIVE: To evaluate the outcome and analyze the methods of surgical treatment of traumatic endophthalmitis following penetrating eye injuries with retained eye foreign bodies . METHODS: A total of 62 consecutive cases (58 men, 4 women) from January 1999 to December 2001 with IOFBs following penetrating eye injuries were retrospectively studied . The ages ranged from 8 to 46 years (mean 23 years) . Sixty patients (63 eyes) underwent pars plana vitreotomy and 1 patient underwent external magnet extraction . The follow-up ranged from 3 to 36 months (mean 12.5 months) . RESULTS: Ten eyes developed endophthalmitis, among which 7 (10.94%) were diagnosed preoperatively . The most frequently cultured organism was Staphylococcus epidermis (44.44%, 4/9) . Postoperatively, retinal detachment due to vitreoretinal proliferation occurred in 5 patients with endophthalmitis and in 9 patients without endophthalmitis . All the retinal detachments were reattached with additional vitreoretinal surgery . Two eyes with endophthalmitis and two without endophthalmitis were eviscerated . CONCLUSIONS: Post-traumatic endophthalmitis with intraocular foreign bodies (IOFBs) deserves great attention because of its high incidence and poor prognosis . Vitrectomy is suggested for the treatment of IOFBs and its complications, and it should be performed as soon as possible . Routine intravenous administration of antibiotics combined with periocular injection and topical antibiotics postoperatively are recommended.

Pediatr Surg Int, 2003 Aug, 19(6), 489 - 94 Epub 2003 May 13.
Small bowel perforation in the premature neonate: congenital or acquired?
Holland AJ, Shun A, Martin HC, Cooke-Yarborough C, Holland J.
To determine the potential aetiological factors of small bowel perforation in the premature neonate, we performed a retrospective chart review of those neonates with spontaneous intestinal perforation (SIP) of the small bowel seen in our tertiary paediatric hospital between January 1980 and December 2000 . Data were collected on gestational feto-maternal health, medical interventions prior to perforation and the subsequent operative and laboratory findings . There were 23 patients with SIP of the small bowel over the 21-year review; 65% were male . There were 7 twin pregnancies but no cases linked to maternal drug abuse . The median gestational age was 27 weeks, the median birth weight 973 g, 19 neonates required ventilation, 15 steroids and 13 indomethacin . The median age at diagnosis was 7 days, heralded by rapid development of abdominal distension in 22 patients . Surgical intervention in addition to insertion of a peritoneal drain was required in 19 patients . Positive microbiological cultures of blood or peritoneal fluid at operation were documented in 8 patients; 5 grew Staphylococcus epidermidis and 4 Candida species . Perforations were located in the ileum in 20 and the jejunum in 1 . Deficiency of the muscularis propria was found in 6 patients . Of the 6 deaths, 2 neonates had significant co-morbidity in addition to extreme prematurity . Small bowel SIP occurs in the premature neonate after the first week of life and usually presents with abdominal distension . Putative risk factors identified included twin gestation, neonatal ventilation, use of steroids and indomethacin, infection with Staphylococcus epidermidis and Candida species and deficiency of enteric smooth muscle.

J Biomater Sci Polym Ed, 2003, 14(4), 313 - 24
Characterization of poly(ethylene oxide) brushes on glass surfaces and adhesion of Staphylococcus epidermidis; Kaper HJ et al.; Poly(ethylene oxide) brushes have been covalently bound to glass surfaces and their presence was demonstrated by an increase in water contact angles from fully wettable on glass to advancing contact angles of 54 degrees, with a hysteresis of 32 degrees . In addition, electrophoretic mobilities of glass and brush-coated glass were determined using streaming potential measurements . The dependence of the electrophoretic mobilities on the ionic strength was analyzed in terms of a softlayer model, yielding an electrophoretic softness and fixed charge density of the layer . Brush-coated glass could be distinguished from glass by a 2-3-fold decrease in fixed charge density, while both surfaces were about equally soft . Adhesion of Staphylococcus epidermidis HBH276 to glass in a parallel plate flow chamber was extremely high and after 4 h, 19.0 x 10(6) bacteria were adhering per cm2 . In contrast, the organisms did not adhere to brush-coated glass, with numbers below the detection limit, i.e . 0.1 x 10(6) per cm2 . These results attest to the great potential of polymer brushes in preventing bacterial adhesion to surfaces.

Ann Med Interne (Paris), 2003 Mar, 154(2), 96 - 100
{Superantigens and vasculitis}; Hamidou M et al.; Superantigens are microbial products, that may activate in a polyclonal way, a high number of T lymphocytes bearing a specific VB segment of TCR; the best characterized are staphylococcal and streptogenic toxins . In a review of the literature, we summarize the role of these superantigens in primitive vasculitis, specially in Kawasaki syndrome and Wegener's granulomatosis . This pathogenic model highlights the complex relations between infection and autoimmunity.

J Neurosurg, 2003 May, 98(5), 1040 - 4
Treatment of staphylococcal ventriculitis associated with external cerebrospinal fluid drains: a prospective randomized trial of intravenous compared with intraventricular vancomycin therapy; Pfausler B et al.; OBJECT: Staphylococcal ventriculitis may be a complication in temporary external ventricular drains (EVDs) . The limited penetration of vancomycin into the cerebrospinal fluid (CSF) is well known; the pharmacodynamics and efficacy of systemically compared with intraventricularly administered vancomycin is examined in this prospective study . METHODS: Ten patients in whom EVDs were implanted to treat intracranial hemorrhage and who were suffering from drain-associated ventriculitis were randomized into two treatment groups . Five of these patients (median age 47 years) were treated with 2 g/day vancomycin administered intravenously (four infusions/day, Group 1), and the other five(median age 49 years) received 10 mg vancomycin intraventricularly once daily (Group 2) . Vancomycin levels were measured in serum and CSF six times a day . The maximum vancomycin level in CSF was 1.73 +/- 0.4 micro/ml in Group 1 and 565.58 +/- 168.71 microg/ml 1 hour after vancomycin application in Group 2 (mean +/- standard deviation) . Vancomycin levels above the recommended trough level of 5 microg/ml in CSF were never reached in Group 1, whereas in Group 2 they below the trough level (3.74 +/- 0.66 microg/ml) only at 21 hours after intraventricular vancomycin application . The vancomycin level in the serum was constant within therapeutic levels in Group 1, whereas in Group 2 in most instances vancomycin was almost below a measurable concentration . In both groups bacteriologically and laboratory-confirmed CSF clearance could be obtained . CONCLUSIONS: Intraventricular vancomycin application is a safe and efficacious treatment modality in drain-associated ventriculitis, with much higher vancomycin levels being achieved in the ventricular CSF than by intravenous administration.

Vet Microbiol, 2003 Jun 24, 94(1), 31 - 8
Cloning of the gene coding for Staphylococcus intermedius exfoliative toxin and its expression in Escherichia coli; Terauchi R et al.; An exfoliative toxin (SIET)-producing strain (D-52) of Staphylococcus intermedius derived from canine pyoderma did not possess large plasmids . Therefore, the gene coding for SIET was considered to be located on the chromosomal DNA . The SIET gene was cloned from the chromosomal DNA of S . intermedius and was expressed in Escherichia coli . The nucleotide sequence of the SIET gene consists of a coding region of 990 bp specifying a polypeptide of 330 amino acid residues, which included a putative 42-residue signal sequence.

Vet Microbiol, 2003 Jun 24, 94(1), 19 - 29
Isolation of exfoliative toxin from Staphylococcus intermedius and its local toxicity in dogs; Terauchi R et al.; A rounding effect was demonstrated in cultured cells inoculated with the culture filtrates (CFs) of 60 strains of Staphylococcus intermedius derived from dogs affected with pyoderma . Exfoliative toxin (ET)-like toxin (ETLT) was isolated from the CF of S . intermedius strain D-52, which exhibited strong rounding activity and then was purified by gel filtration on a Sephadex G-75 column, and by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) . The ETLT caused exfoliation in 1-day-old chickens, suckling Syrian hamsters, and dogs, but not in suckling mice . The ETLT was serologically different from exfoliative toxin A (ETA), exfoliative toxin B (ETB), exfoliative toxin C (ETC), S . hyicus exfoliative toxin A (SHETA), and SHETB, as shown by Western blot analysis . The molecular weight of the ETLT was estimated at 30 kDa by SDS-PAGE . In the present study, we propose the ETLT was a novel type of ET, S . intermedius exfoliative toxin (SIET).

Curr Opin Infect Dis, 2003 Apr, 16(2), 71 - 6
Staphylococcal epidermolysins; Prevost G et al.; PURPOSE OF REVIEW: Staphylococcal epidermolysins are the major causative toxins of bullous impetigo and staphylococcal scalded skin syndrome . This disease is characterized by the splitting of the epidermis between two cell layers resulting in exfoliation . It predominantly affects newborn babies and exposes them to secondary infections . This leads to the risk of epidemics, especially in nurseries . With only an experimental model which consists of skin injections in newborn mice and the recent determination of three-dimensional structures, the essential function of these toxins remained controversial, split between that of specific proteases and that of superantigens . RECENT FINDINGS: Staphylococcal epidermolysins now constitute a family of toxins, with the recent characterizations of two new serotypes: ETC and ETD . They may be secreted by sensitive or methicillin-resistant strains . Four molecules were also identified in Staphylococcus hyicus responsible for exudative epidermitis in swine . While different observations suggested a proteolytic action to these toxins, the histological parallel made with pemphigus foliaceus greatly helped in the characterization of the targets for epidermolysins ETA, ETB, ETD: desmoglein-1, a desmosome-constitutive protein, and incidentally melanocyte-stimulating hormones, which accounts for the blisters observed clinically . SUMMARY: The growing complexity in staphylococcal toxins has to be taken into account both for their association with diseases and for diagnosis purposes . Even though cases of staphylococcal scalded skin syndrome in adults are rare, they raise further questions about the pathogenic features of the disease such as individual sensitivity and distribution of the toxins into the body.

J Immunol, 2003 May 15, 170(10), 5159 - 64
Toll-like receptor 2 (TLR2) and TLR9 signaling results in HIV-long terminal repeat trans-activation and HIV replication in HIV-1 transgenic mouse spleen cells: implications of simultaneous activation of TLRs on HIV replication; Equils O et al.; Opportunistic infections are common in HIV-infected patients; they activate HIV replication and contribute to disease progression . In the present study we examined the role of Toll-like receptor 2 (TLR2) and TLR9 in HIV-long terminal repeat (HIV-LTR) trans-activation and assessed whether TLR4 synergized with TLR2 or TLR9 to induce HIV replication . Soluble Mycobacterium tuberculosis factor (STF) and phenol-soluble modulin from Staphylococcus epidermidis induced HIV-LTR trans-activation in human microvessel endothelial cells cotransfected with TLR2 cDNA . Stimulation of ex vivo spleen cells from HIV-1 transgenic mice with TLR4, TLR2, and TLR9 ligands (LPS, STF, and CpG DNA, respectively) induced p24 Ag production in a dose-dependent manner . Costimulation of HIV-1 transgenic mice spleen cells with LPS and STF or CpG DNA induced TNF-alpha and IFN-gamma production in a synergistic manner and p24 production in an additive fashion . In the THP-1 human monocytic cell line stably expressing the HIV-LTR-luciferase construct, LPS and STF also induced HIV-LTR trans-activation in an additive manner . This is the first time that TLR2 and TLR9 and costimulation of TLRs have been shown to induce HIV replication . Together these results underscore the importance of TLRs in bacterial Ag- and CpG DNA-induced HIV-LTR trans-activation and HIV replication . These observations may be important in understanding the role of the innate immune system and the molecular mechanisms involved in the increased HIV replication and HIV disease progression associated with multiple opportunistic infections.

J Med Primatol, 2003 Feb, 32(1), 23 - 30
Flow cytometric analysis of macaque whole blood for antigen-specific intracellular cytokine production by T lymphocytes; Keeney TS et al.; We report here the standardized conditions for stimulation of macaque whole blood samples with various protein or peptide antigens, and production of significant intracellular levels of interferon gamma (IFN-gamma) and tumor necrosis factor alpha (TNF-alpha) in CD4+ as well as CD8+ T lymphocytes . We observed significantly higher levels of TNF-alpha compared with IFN-gamma in both CD4+ and CD8+ T lymphocytes from all the macaque whole blood samples stimulated with staphylococcal enterotoxin B (SEB) as an antigen . Similarly, when whole blood samples from rhesus macaques immunized with an HIV envelope peptide cocktail vaccine were stimulated with either the peptide cocktail or recombinant gp160, we observed production of significant levels of TNF-alpha by both CD4+ and CD8+ T lymphocytes . These results strongly support the utility of the whole blood cytokine flow cytometry methodology for determining antigen-specific immune responses of macaques in vaccine studies.

J AOAC Int, 2003 Mar-Apr, 86(2), 332 - 9
First proficiency testing to evaluate the ability of European Union National Reference Laboratories to detect staphylococcal enterotoxins in milk products; Hennekinne JA et al.; The European Commission has designed a network of European Union-National Reference Laboratories (EU-NRLs), coordinated by a Community Reference Laboratory (CRL), for control of hygiene of milk and milk products (Council Directive 92/46/ECC) . As a common contaminant of milk and milk products such as cheese, staphylococcal enterotoxins are often involved in human outbreaks and should be monitored regularly . The main tasks of the EU-CRLs were to select and transfer to the EU-NRLs a reference method for detection of enterotoxins, and to set up proficiency testing to evaluate the competency of the European laboratory network . The first interlaboratory exercise was performed on samples of freeze-dried cheese inoculated with 2 levels of staphylococcal enterotoxins (0.1 and 0.25 ng/g) and on an uninoculated control . These levels were chosen considering the EU regulation for staphylococcal enterotoxins in milk and milk products and the limit of detection of the enzyme-linked immunosorbent assay test recommended in the reference method . The trial was conducted according to the recommendations of ISO Guide 43 . Results produced by laboratories were compiled and compared through statistical analysis . Except for data from 2 laboratories for the uninoculated control and cheese inoculated at 0.1 ng/g, all laboratories produced satisfactory results, showing the ability of the EU-NRL network to monitor the enterotoxin contaminant.

Curr Microbiol, 2003 Apr, 46(4), 280 - 6
Cell surface hydrophobicity and slime production of Staphylococcus epidermidis Brazilian isolates; Krepsky N et al.; The cell surface hydrophobicity of 60 isolates and three reference strains of Staphylococcus epidermidis was assayed by means of bacterial aggregation in liquid broth, phosphate-buffered saline, and in ammonium sulfate, as well as by affinity of the bacteria to n-hexadecane and polystyrene surfaces . In order to better characterize the isolates, the influence of bacterial growth time and enzyme treatment on cell hydrophobicity and the analysis of the slime production were also investigated . The strains presented the following profiles when assayed by the ammonium sulfate aggregation test (SAT): SAT < 1M, SAT 1M - <2M, SAT 2M - <4M, and SAT >or=4M . When SAT < 1M, the strains showed positive results for most of the cell surface hydrophobicity tests . None of the strains belonging to the groups with SAT >or= 1M showed spontaneous aggregation (SA), auto-aggregation (AA), or glass adherence, albeit 32 (62.7%) strains were polystyrene adherent and 42 (82.3%) presented weak adherence to n-hexadecane (>20%) . The best correlation of the results was found among the AA and glass adherence tests (100%), followed by SA/ glass adherence (98%) and SA/ AA test (98%) . The polystyrene adherence test and microbial adherence to n-hexadecane test (MATH) showed 78% correlation . Proteinase K treatment reduced bacterial adherence to polystyrene, but did not influence the SAT values . Three distinct groups of strains were distinguished by the polystyrene micromethod and glass tube adherence assay: 0.0-0.4 O.D . group, including non-glass adherent isolates; 0.5-0.7 O.D . group, including strains with variable profiles (adherent or non-adherent); and 0.8-1.3 O.D . group, composed of glass-adherent strains . Evaluation by a single method seemed not to reliably determine the surface hydrophobicity characteristics of S . epidermidis clinical isolates . Auto-aggregation properties of the strains that adhered to glass seemed related to slime expression, rather than cell surface hydrophobicity . Data also suggested involvement of protein components in adherence to polystyrene, but not in auto-aggregation properties assayed by SAT.

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