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 medi