Klin Mikrobiol Infekc Lek, 2004 Dec, 10(6), 259 - 264 {Microbiological findings in children with an indwelling central venous catheter.}; Uher M et al.; Introduction: Besides their obvious advantages for the patient, central venous catheters (CVC) also carry the risk of possible infectious complications . The purpose of our investigation was to carry out a microbiological evaluation of a 5-year set of paediatric patients with indwelling CVC.s Patients and methods: In the group were 218 CVCs inserted to 165 children over a period of 5 years . There were 26 multi-lumen catheters (11.927 %) and 192 single-lumen catheters (88.073 %) . The mean indwelling period was 10.1 days per 1 CVC . Blood for microbiology was removed by a physician from the CVC after disinfecting its opening under standard sterile conditions into a commercial sampling vessel HEMOD (Imuna, Sarisske Michalany, Slovak Republic) or into a vessel of an automated haemoculture system BactecPeds PLUS/F (Becton Dickinson and Comp., Spark,MA, USA) . When removing the tip of the CVC we disinfected, before removing the CVC, the area around the insertion with isopropyl or ethyl alcohol . We released the fixed CVC and 1 minute after disinfection we pulled out the CVC and cut off the end or rather the tip of the catheter (approx . 1-3 cm of the tip) into a sterile test tube . To establish the diagnosis of infectious complications we used the 1995 Sirges-Serra classification and the CDC criteria . Results: In 5 years (1995-1999) we had 71 infectious complications . There were 31 contaminated catheters, 27 cases of catheter sepsis and 11 cases of catheter bacteraemia . With 147 catheters (67.43%) there were no infectious complications . Dominant microbes were Staphylococcus epidermidis (32 cases - 11 from haemocultures and 21 from CVCs) and Candida spp . (30 cases, 17 from haemocultures and 13 from CVCs) . Among the microbiological agents of catheter sepsis predominated Gram-negative bacteria . Out of the whole analysed group 41 children (24.8 %) died . CVC as the cause of death was demonstrated in 6 children (3.636 % of patients with CVC) . Conclusions: Microbiological findings in our group are in line with literary data . To reduce the incidence of infectious complications it is important to limit sampling from CVC to a minimum . Insertion of CVCs under strict sterile conditions and aseptic handling of all entries into the central bloodstream reduces to a minimum the risk of infectious complications. Clin Exp Immunol, 2005 Feb, 139(2), 189 - 201 Correlation of disease evolution with progressive inflammatory cell activation and migration in the IL-4 transgenic mouse model of atopic dermatitis; Chen L et al.; Summary Atopic dermatitis is a chronic inflammatory skin disease characterized by inflammatory cell infiltration in the skin . In order to assess the roles of inflammatory cells in this disease, we analysed the activation status and surface markers of various leucocytes in the IL-4 transgenic mouse model of atopic dermatitis, by flow cytometry, immuofluorescence microscopy, and T cell proliferation assays . The studies were performed with a nontransgenic mouse control and transgenic mice at three disease stages: before disease onset, early skin disease, and late skin disease, so that we can delineate the immunological sequence of events . As the skin disease evolves, the skin draining lymph node cells from IL-4-Tg mice show a spontaneous proliferation and a progressively enhanced proliferative response to stimulants including anti-CD3, Con A, PHA, and Staphylococcus enterotoxins A and B . As the disease evolves, the percent of lymphoid organ T cells expressing activation molecules (CD44 and CD69) and costimulatory molecules (ICOS and PD-1) are progressively increased; the percent and total number of T cells are reduced in an incremental manner in the secondary lymphoid organs while the number of T cells infiltrating the skin increases in an incremental fashion; the total number of dendritic antigen presenting cells, macrophages, and NK cells gradually increases in the lymphoid organs . Collectively, our results suggest that there is a continued and progressive migration of activated inflammatory cells from the secondary lymphoid organs into the skin where they participate in immune responses resulting in the pathology associated with inflammation. Blood . 2005 Jan 11; {Epub ahead of print} Superantigen recognition by HLA class II on monocytes up-regulates toll-like receptor 4 and enhances proinflammatory responses to endotoxin; Hopkins PA et al.; The devastating systemic effects of bacterial superantigens may be explained by powerful proinflammatory synergy with lipopolysaccharide (LPS) . However, the mechanism underlying this phenomenon remains unclear and has never been investigated in humans . Specifically, there is no known link between superantigen-induced immune effects and the pattern-recognition of LPS at toll-like receptor 4 (TLR4) . Here we show that bacterial superantigens induce rapid transcription and increased membrane expression of TLR4 in primary human monocytes by ligation of MHC class II . We also demonstrate that superantigens are solely responsible for monocyte TLR4 up-regulation induced by products from Gram-positive bacteria . In parallel with enhanced TLR4 expression, priming of purified monocytes or mixed peripheral blood mononuclear cells with superantigens significantly enhanced the induction of proinflammatory cytokines by known TLR4 ligands . Staphylococcal enterotoxin A constructs containing targeted mutations were used to demonstrate a requirement for MHC class II ligation in both TLR4 up-regulation and enhanced responses to endotoxin . In contrast to results from animal models, superantigen-endotoxin interaction was not dependent on T cell receptor ligation by superantigen or interferon gamma production . Pattern-recognition of bacterial superantigens by MHC class II receptors may exacerbate the proinflammatory response of monocytes to Gram-negative infection or endotoxin by up-regulation of TLR4. Vet Ophthalmol, 2005 Jan, 8(1), 33 - 37 Survey of bacterial microorganisms in the conjunctival sac of clinically normal dogs and dogs with ulcerative keratitis in Fortaleza, Ceara, Brazil; Prado MR et al.; Objective The ocular microflora in dogs has not been established in north-east Brazil . Thus, the main aim of this research was to determine the bacterial microorganisms in the conjunctival sac of clinically normal dogs and dogs with ulcerative keratitis in Fortaleza, Ceara, Brazil . Animals studied This study included 60 healthy dogs, 15 dogs with unilateral corneal ulcer, and three dogs with bilateral corneal ulcers . Procedure Samples were taken by a calibrated platinum loop (1 microL) placed directly onto the conjunctival sac and on sterile blood agar . The clinical specimens were incubated at 37 degrees C in an atmosphere of 5% CO(2) for 48 h . Results Of the 120 samples from healthy dogs, only 47 (39%) had positive culture for bacteria, while all of the specimens from eyes with corneal ulcer were positive for bacterial growth . The group of dogs with corneal ulcer had a higher (P < 0.05) number of colony-forming units (CFU) per plate than the group of healthy animals . Of the 59 isolates from healthy eyes, only nine (15.3%) had more than 50 CFU per plate, while in the group of dogs with corneal ulcer, 23 (62.2%) of the 37 isolates presented more than 50 CFU per plate . In both groups Gram-positive bacteria (86.5%) predominated over Gram-negative (13.5%) . Staphylococcus spp . was the most frequently isolated genus and S . intermedius predominated in both groups . Conclusion The results of our study are directly applicable to initiate rational, preventive and therapeutic measures with greater accuracy in dogs with corneal ulcer. Am J Transplant, 2005 Jan, 5(1), 183 - 8 Mycoplasma hominis Septic Arthritis in a Pediatric Renal Transplant Recipient: Case Report and Review of the Literature; Mian AN et al.; Septic arthritis (SA) typically occurs in young children, often from Staphylococcus . With chronic immunosuppression, however, pathogens may be atypical . A 15-year-old African-American female developed Mycoplasma hominis SA in her right hip 2 months following cadaveric renal transplant (Tx) . Her presentation was subtle and indolent, without fever or leukocytosis . Although reported in adult Tx recipients, M . hominis infections have not been described in pediatric recipients . Early immunosuppression (basiliximab, prednisone, tacrolimus, mycophenolate mofetil and Thymoglobulin) may have increased her susceptibility to M . hominis . Optimal therapy for M . hominis SA is not well established and relapses occur . This patient underwent joint incision and drainage, treatment for 8 weeks with doxycycline and levofloxacin guided by in vitro sensitivities, and a reduction in immunosuppression . She has been free of ongoing infection for 3 years with stable graft function (Cr 1.1 mg/dL) on moderate immunosuppression with prednisone, tacrolimus and MMF. Zh Mikrobiol Epidemiol Immunobiol, 2004 Nov-Dec, (6), 60 - 3 {Activation of Langat virus infection in mice under the influence of Licopid}; Mutation Analysis of the Histidine Residues in the Glycylglycine Endopeptidase ALE-1; Department of Bacteriology, Hiroshima University Graduate School of Biomedical Sciences, Kasumi 1-2-3, Minami-ku, Hiroshima 734-8553, Japan . sugai@hiroshima-u.ac.jpA novel staphylolytic enzyme, ALE-1, is a glycylglycine endopeptidase produced by Staphylococcus capitis EPK1 . ALE-1 possesses seven histidines . Chemical modification studies using diethylpyrocarbonate and iodoacetic acid suggested that a histidine or tyrosine residue(s) in the molecule is important for the organism's staphylolytic activity . All of the histidine residues, one tyrosine, and one aspartic acid residue in the N-terminally truncated ALE-1 (DeltaN-term ALE-1) were systematically altered by site-directed mutagenesis, and the enzyme activities and metal contents of the variants were measured . Our studies indicated that His-150, His-200, His-231, His-233, and Asp-154 are essential for the enzyme activity of DeltaN-term ALE-1 . Except for His-150 and Asp-154, all of these amino acids were located within the 38-amino-acid region conserved among 11 proteins, including 5 staphylolytic endopeptidases . Inductively coupled plasma-mass spectrometric analysis of DeltaN-term ALE-1 revealed that it contains one atom of zinc per molecule . Measurement of the zinc content of the mutant DeltaN-term ALE-1 suggested that His-150 and -233 are important for zinc binding; their loss in these variant enzymes coincided with the loss of staphylolytic activity . These results strongly suggest that ALE-1 is a novel member of zinc metalloproteases. Br J Plast Surg, 2005 Jan, 58(1), 73 - 80 Lyell syndrome revisited: analysis of 18 cases of severe bullous skin disease in a burns unit; Neff P et al.; Over the last few years, understanding of the pathophysiology of toxic epidermal necrolysis (TEN), or Lyell's disease, has substantially increased . However, differentiation of severe bullous skin disease remains a challenge for the clinician, and one that is often complicated by late patient referral . We performed a retrospective analysis of all patients with severe bullous skin disease, admitted between 1997 and 2002 to the Burn Centre, which is an integrated part of the Division for Plastic, Hand- and Reconstructive Surgery at the University Hospital of Zurich, Switzerland . We present an overview of our strategies and of the diagnostic and therapeutic difficulties encountered . The final diagnoses of the 18 patients referred to the unit were as follows: eight cases of TEN, one case of staphylococcal scalded-skin syndrome (SSSS), two cases of generalised drug eruption, one case of acute generalised exanthematic pustulosis and one case of febrile ulceronecrotic pityriasis lichenoides et varioliformis acuta (PLEVA) . In two cases, the diagnosis remained unclear . In three cases, paraneoplastic origins were suspected but not demonstrated . The overall mortality rate was 33% (six of 18 patients) . Remarkably, all patients with histologically confirmed TEN survived . Six of these patients were successfully treated with intravenous immunoglobulins (IVIG) . The most common single causative drug inducing TEN (four cases out of eight) was Phenytoin . Establishing an accurate diagnosis-based on a skin biopsy, harvested at an early stage-is more important than ever, because more specific and effective therapeutic modalities are available . As these potentially life-threatening bullous skin disorders are rare, we recommend, that care be provided by an experienced interdisciplinary team, comprising a dermatologist, or dermatopathologist, an intensive care specialist and a plastic surgeon. Arch Biochem Biophys, 2005 Feb 1, 434(1), 86 - 92 Probing the subtle conformational state of N138ND(2)-Q106O hydrogen bonding deletion mutant (Asn138Asp) of staphylococcal nuclease using time of flight mass spectrometry with limited proteolysis; Huang S et al.; Recent studies indicate that the N138ND(2)-Q106O hydrogen bonding deletion in staphylococcal nuclease significantly alters the conformational integrity and stability of the nuclease . To find out the structural basis of the changes, mass spectrometry and limited proteolysis methods were combined to probe the subtle conformational changes in the SNaseN138D mutant and SNaseN138D-Ca(2+)-pdTp complex . The results reveal that the N138ND(2)-Q106O hydrogen bonding deletion makes the C-terminal part of alpha-helix 1 and alpha-helix 2 in the C-terminal subdomain of SNaseN138D unfold to some extent, but does not have much effect on the N-terminal part of alpha-helix 1, alpha-helix 3, and the N-terminal beta-barrel subdomain of SNaseN138D . Binding of ligands makes the alpha-helices 1 and 2 more resistant to protease Glu-C attack and converts the partially unfolded state to a native-like state . This study also demonstrates how mass spectrometry can be combined with limited proteolysis to observe conformational changes induced by ligand binding. Inflamm Bowel Dis, 2004 Nov, 10(6), 801 - 10 Effects of infliximab on apoptosis and reverse signaling of monocytes from healthy individuals and patients with Crohn's disease; Ringheanu M et al.; OBJECTIVES: Infliximab, an anti-tumor necrosis factor (TNF) monoclonal antibody, might exert some of its long-term therapeutic effects in Crohn's disease (CD) by interacting directly with cells of the immune system such as monocytes and T lymphocytes via membrane TNF and by inducing apoptosis . Accordingly, the effects of inflix-imab on monocyte apoptosis and down-regulation of proinflammatory cytokines (reverse signaling) were assessed . METHODS: To assess apoptosis, monocytes from healthy individuals (controls) and CD patients were incubated in the presence or absence of infliximab or the apoptotic agent gliotoxin for 24 hours . Annexin V staining and the terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-FITC nick end labeling assay were used to measure early and late apoptosis . To measure the effects of infliximab on reverse signaling, monocytes from healthy individuals pretreated in vitro with infliximab were stimulated with lipopolysaccharide or staphylococcal enterotoxin A, and the induction of the proinflammatory cytokines, TNF-alpha, interleukin (IL)-1beta, IL-6, and IL-8 was measured by reverse transcription polymerase chain reaction . The effect of in vivo infliximab treatment of monocytes was similarly determined by comparing the responses of monocytes from CD patients before and immediately after infliximab infusion . RESULTS: Infliximab did not induce apoptosis of monocytes from either healthy individuals or CD patients but rather stabilized them . However, monocytes from healthy individuals treated with infliximab in vitro, or from CD patients infused with infliximab, produced significantly less TNF and other proinflammatory cytokines when stimulated with the bacterial products lipopolysaccharide and staphylococcal enterotoxin A . CONCLUSIONS: Apoptosis of monocytes is not responsible for the therapeutic effects of infliximab . However, some of the therapeutic effects of infliximab may be caused by its ability to down-regulate proinflammatory cytokines production by monocytes exposed to bacterial antigens. J Biochem (Tokyo), 2004 Oct, 136(4), 427 - 431 Essential Residues, W177 and R198, of LukF for Phosphatidylcholine-Binding and Pore-Formation by Staphylococcal {gamma}-Hemolysin on Human Erythrocyte Membranes; Monma N et al.; LukF and Hlg2 of staphylococcal gamma-hemolysin assemble into hetero-oligomeric pores on human red blood cells (HRBC) . Here, we demonstrate, using a single-molecule imaging technique, that a W177T/R198T mutant of LukF, which exhibits no binding activity toward phosphatidylcholine, could form intermediate oligomers with Hlg2, including dimers, tetramers, and hexamer/heptamers, on HRBC . But, the mutant neither caused K(+) efflux nor lysed HRBC, indicating that functional pores were not formed . Hence, we conclude that the W177 and R198 residues are essential for proper pore-formation by staphylococcal gamma-hemolysin . We also suggest that the interaction between the W177 and R198 residues, and phosphatidylcholine on membranes is the key to the formation of functional pores. Infect Immun, 2005 Jan, 73(1), 50 - 61 Role of CD44 and its v7 isoform in staphylococcal enterotoxin B-induced toxic shock: CD44 deficiency on hepatic mononuclear cells leads to reduced activation-induced apoptosis that results in increased liver damage; McKallip RJ et al.; Exposure to bacterial superantigens such as staphylococcal enterotoxin B (SEB) leads to the induction of toxic shock syndrome which results in multiorgan failure, including liver damage . In the present study, we investigated the role of CD44 in SEB-induced liver injury . Injection of SEB into d-galactosamine-sensitized CD44 wild-type (WT) mice led to a significant increase in CD44 expression on liver T cells, NK cells, and NKT cells . Administration of SEB to CD44 knockout (KO) mice caused significantly enhanced liver damage which correlated with elevated numbers of T cells, NK cells, NKT cells, and macrophages in the liver and increased production of tumor necrosis factor alpha and gamma interferon compared to CD44 WT mice . Furthermore, liver mononuclear cells from CD44 KO mice were resistant to SEB-induced apoptosis, and cDNA microarray analysis revealed that SEB activation of such cells led to the induction of several antiapoptotic genes and repression of proapoptotic genes . Examination of CD44 isoforms revealed that SEB exposure altered CD44 variant 7 (v7) isoform expression . Interestingly, mice bearing a specific deletion of the CD44v7 exon exhibited increased susceptibility to SEB-induced hepatitis . Finally, treatment of CD44 WT mice with anti-CD44 monoclonal antibodies reduced expression of CD44 in liver mononuclear cells and caused increased susceptibility to SEB-induced liver injury . Together, these data demonstrate that the expression of CD44 and/or CD44v7 on SEB-activated liver mononuclear cells facilitates their rapid apoptosis, thereby preventing severe liver injury in wild-type mice, and suggest that CD44 plays an important role in the regulation and elimination of immune cells in the liver. J Cataract Refract Surg, 2004 Dec, 30(12), 2638 - 40 Early bacterial keratitis after laser-assisted subepithelial keratectomy; Laplace O et al.; We report a case of bacterial keratitis that occurred after laser-assisted subepithelial keratectomy . The patient presented with a decrease in visual acuity and pain 2 days after the procedure . Culture was positive for Staphylococcus haemolyticus . The infiltrate slowly resolved with topical antibiotics, and the best corrected visual acuity improved to 20/20 . Although bacterial keratitis occurs rarely after refractive surgery, patients should be informed of the potential risk for visual loss caused by this infection. Presse Med, 2004 Dec 4, 33(21), 1511 - 5 {Acute aseptic meningitis in adults . Retrospective study of 32 cases}; Pavic M et al.; OBJECTIVE: The objective of this study was to analyze the characteristics of a series of acute aseptic meningitis (AAM) (defined by sterile cerebrospinal fluid (CSF) with more than 10 leucocytes per mm3 and a neutrophilic polynuclear-rich formula) . We analysed the initial management, the reasons for antibiotic and/or antiviral treatment, the aetiologies, the need for lumbar puncture and the progression.. . METHOD: We retrospectively analyzed 32 cases of AAM (out of a total of 130 cases of meningitis) from two departments of internal medicine in Lyon, diagnosed between January 1996 and January 2003 . Only the files fulfilling the AAM criteria were retained, selecting those with a minimum neutrophilic polynuclear level of 30% in the CSF . RESULTS: The mean age was 32.6 years (range: 18-75) and predominantly male patients (59%) . On admission, 87% of the patients exhibited fever, but only 9% remaining so for 72 hours . Viral syndrome before admission was noted in 59% of cases, with seasonal predilection (summer: 39%, winter: 35%) . The motivation for lumbar puncture (LP) was meningeal syndrome (44%), headache (94%) and vomiting (47%) . The average rate of neutrophils in the CSF on admission was 63% (range: 30-96) . A control LP on Day 3 was performed 16 times (50%): mean PNN rate at 18% (range: 0-80), lymphocyte rate=68% (range: 20-95) . Most of the patients (77.4%) had a C reactive protein (CRP) lower than 50 mg/l on admission (range: 5-320) . A cerebral scan was performed 10 times (31%) and was abnormal 2 times (multiple cerebral abscesses, possible intracranial hypertension) . An antibiotic (84%) and/or antiviral (34%) treatment was initiated . The evolution on Day 3 was favourable (87.5%): no fever, regression of the meningeal syndrome, with a mean duration of hospitalisation of 8.3 days (range: 1-60) . Search for Herpes simplex virus and Enterovirus was made with PCR analysis in 20 cases (62.5%): no positivity for the herpes, but 9 for the Enterovirus . The systematic blood cultures were positive only once (staphylococcal infective endocarditis with cerebral abscesses) . The diagnosis of bacterial meningitis was evoked 3 times (prior antibiotic treatment) . DISCUSSION: The frequency of Enterovirus AAM should encourage this type of investigation in order to withdraw the often initiated anti-infectious treatment rapidly, and hence avoid a second lumbar puncture. Syst Appl Microbiol, 2004 Nov, 27(6), 696 - 702 Rapid and reliable identification of Staphylococcus equorum by a species-specific PCR assay targeting the sodA gene; Blaiotta G et al.; Rapid and reliable identification of Staphylococcus (S.) equorum was achieved by species-specific PCR assays . A set of primers targeting the manganese-dependent superoxide dismutase (sodA) gene of S . equorum was designed . Species-specificity of the primer set was evaluated by using a total of 112 strains (including 27 reference strains of the DSM collection), representing 26 different species of the genus Staphylococcus, 3 species of the genus Kocuria, and different strains of Macrococcus caseolyticus . By using primers SdAEqF and SdAEqR the expected PCR fragment was obtained only when DNA from S . equorum strains was used as template . 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 . equorum strains. Vet Immunol Immunopathol, 1980 Feb, 1(2), 179 - 93 Enzyme linked immunoassay and fluorescent antibody techniques in the diagnosis of viral diseases using staphylococcal protein-A instead of anti-gamma-globulins; Bommeli W et al.; Staphylococcal protein-A (SpA) is known to interact with the crystallizable fragment (Fc) of IgG molecules from several species . In the present study, SpA coupled to either fluorescein isothiocyanate (FITC) or peroxidase was used in place of antisera to IgG for the fluorescent antibody (FA) techniques and the enzyme linked immunoassay (ELISA) . The SpA conjugates produced low background staining when applied in these techniques, and provide a rapid, highly specific and sensitive means for the identification of viral isolates and the detection of serum antibodies . Moreover, SpA is a single reagent that replaces various preparations of anti-gamma globulin against many species . SpA-FITC conjugate was successfully applied for the identification of pseudorabies virus, hog cholera virus, swine vesicular disease virus, transmissible gastroenteritis virus, porcine parvovirus and porcine enteroviruses . Antibody titers against the mentioned viruses could be determined semi-quantitatively in the indirect FA test with SpA-FITC . In our laboratory the ELISA became a routinely practicable serological test for the detection of antibodies only after we introduced SpA-peroxidase as a marker for the IgG. Eur J Biochem, 2004 Dec, 271(23-24), 4815 - 24 Solution structure of the active-centre mutant I14A of the histidine-containing phosphocarrier protein from Staphylococcus carnosus; Moglich A et al.; High-pressure NMR experiments performed on the histidine-containing phosphocarrier protein (HPr) from Staphylococcus carnosus have shown that residue Ile14, which is located in the active-centre loop, exhibits a peculiarly small pressure response . In contrast, the rest of the loop shows strong pressure effects as is expected for typical protein interaction sites . To elucidate the structural role of this residue, the mutant protein HPr(I14A), in which Ile14 is replaced by Ala, was produced and studied by solution NMR spectroscopy . On the basis of 1406 structural restraints including 20 directly detected hydrogen bonds, 49 (1)H(N)-(15)N, and 25 (1)H(N)-(1)H(alpha) residual dipolar couplings, a well resolved three-dimensional structure could be determined . The overall fold of the protein is not influenced by the mutation but characteristic conformational changes are introduced into the active-centre loop . They lead to a displacement of the ring system of His15 and a distortion of the N-terminus of the first helix, which supports the histidine ring . In addition, the C-terminal helix is bent because the side chain of Leu86 located at the end of this helix partly fills the hydrophobic cavity created by the mutation . Xenon, which is known to occupy hydrophobic cavities, causes a partial reversal of the mutation-induced structural effects . The observed structural changes explain the reduced phosphocarrier activity of the mutant and agree well with the earlier suggestion that Ile14 represents an anchoring point stabilizing the active-centre loop in its correct conformation. Curr Opin Drug Discov Devel, 2004 Mar, 7(2), 248 - 56 Engineering protein A affinity chromatography; Jungbauer A et al.; Staphylococcal protein A can selectively interact with immunoglobulins . This protein is widely used as a ligand for affinity chromatography to purify therapeutic antibodies on an industrial scale . This type of affinity chromatography constitutes a generic step in processing antibodies . Questions of scale-up, design of chromatographic conditions, clearance of adventitious agents and operational modes, such as continuous operation or purification of antibodies in expanded-bed mode, will be addressed in this review. Curr Opin Drug Discov Devel, 2004 Mar, 7(2), 223 - 7 MSCRAMM--targeted vaccines and immunotherapy for staphylococcal infection; Rivas JM et al.; Hospital-acquired infections are associated with prolonged hospitalization and an increase in both healthcare costs and resources . Advances in sophisticated medical procedures, an increase in the number of immunocompromised patients, and the continued emergence of resistance to conventional antibiotic therapy has created a need for alternative strategies to prevent and treat infectious bacterial diseases . Immunoprevention and immunotherapy targeting microbial surface components recognizing adhesive matrix molecule (MSCRAMM) proteins are viable approaches to potentially impede bacterial adherence, eliminate colonization, and minimize hematogenous dissemination, thereby halting the inception and progression of infection . This review summarizes several investigative efforts where staphylococcal MSCRAMM proteins are being utilized in the design of subunit vaccines and in the development of innovative therapeutic strategies that could be implemented following the onset of infection to manage severe and life-threatening disease. J Proteome Res, 2004 Nov-Dec, 3(6), 1278 - 83 2D LC/MS analysis of membrane proteins from breast cancer cell lines MCF7 and BT474; Xiang R et al.; Membrane proteins play a central role in the interaction of the cell with its environment and in the function of subcellular organelles . The current study focused on developing a better understanding of the membrane proteome of two well-characterized breast cancer cell lines . Membranes from osmotically lysed BT474 and MCF7 cells were treated with cyanogen bromide followed by a combination of trypsin and Staphylococcus V8 protease to obtain hydrophilic peptides from membrane proteins . The complex peptide mixtures obtained were separated by 2-dimensional liquid chromatography coupled online with a nano-electrospray ionization ion trap mass spectrometer (2D LC/nanoESI-MS) . The strong cation exchange column used in the first dimension of the separation was eluted in an automated fashion using a series of salt steps of increasing concentration . Peptides eluted from each of the salt steps were separated using a capillary reversed-phase HPLC column, the output of which was directed through a nano-electrospray fused silica tip into the mass spectrometer . Peptides were fragmented by collision-induced dissociation (CID) and analyzed by data-dependent MS/MS followed by database searching using the Sequest algorithm . Analysis of the data revealed both similarities and expected differences between proteins identified from these cell lines . As demonstrated by others, mRNA and the HER2/neu protein tyrosine kinase-linked receptor in BT474 cells is up regulated compared to its level in MCF7, while the expression of the estrogen receptor alpha is known to be up regulated in MCF7 cells . As expected, our studies showed identification of peptides from HER2 in BT474 while estrogen receptor peptides were detected in the MCF7 line . A total of 604 proteins were identified from BT474 membranes while 313 proteins were found from MCF7 . The results are discussed in terms of the known differences in both protein and mRNA expression between these two breast cancer cell lines and also in the context of other known phenotypic differences between these cells. Arch Virol . 2004 Dec 10; {Epub ahead of print} Therapeutic effects of dengue 2 virus capsid protein and staphylococcal nuclease fusion protein on dengue-infected cell cultures; Qin CF et al.; Dengue infection poses a serious public health problem in most tropical and subtropical areas . No effective antiviral drugs or vaccines are currently available against dengue infection . To explore the feasibility of using capsid-targeted viral inactivation (CTVI) as an antiviral strategy against dengue infection, we constructed a plasmid expressing a fusion protein consisting of staphylococcal nuclease (SN) fused to dengue 2 virus capsid protein (D2C), and investigated its effects on the production of infectious virions when introduced into BHK cells infected with dengue virus . The results indicated that D2C-SN can be expressed and tolerated in this mammalian cell culture . The enzymatically active SN moiety was incorporated into nascent virions during the process of viral assembly . By comparing the effects of incorporated SN and SN*, an enzymatically inactive missense mutant form of wild-type SN, on the infectivity of progeny virions, we clearly demonstrated that nucleolytic activity was the major antiviral mechanism . Expression of D2C-SN fusion protein as a therapeutic agent resulted in a reduction in infectious titers of 12- to 60-fold . Therefore, dengue virus may be particularly vulnerable to a CTVI therapeutic approach. Cell Tissue Bank, 2004, 5(4), 261 - 6 Processing of cardiovascular allografts: effectiveness of European Homograft Bank (EHB) antimicrobial treatment (cool decontamination protocol with low concentration of antibiotics); Tabaku M et al.; To assess the effectiveness of antimicrobial treatment by using cool decontamination protocol with low concentration of antibiotics during processing of cardiovascular allografts, 948 allografts processed during a 2-year period were analysed . Five hundred and fourty one donors aged <62 years were classified in: multiorgan donors (MOD) with non-transplantable hearts; recipients of cardiac transplantation (RHT); and non-beating heart cadavers with a warm ischemic time of less than 6 h (NBHD) . During processing three samples for bacteriology testing were taken A (sampling before decontamination); B (sampling after decontamination); C (sampling on the final product) . Samples A were positive in 348 cases (36.4%), respectively 36% for MOD, 21.6% for RHT and 78.1% for NBHD . All the allografts were immersed in a cocktail of four antibiotics at 4 degrees C . After exposure to antibiotics the rate of decontamination of those with A positive was 90.4, 92.5, 82.5% respectively for MOD, RHT, NBHD . At the end of processing, 57 allografts (6%) were positive in B and/or C, 15 allografts remained contaminated with the same bacteria as in A, 42 were contaminated during processing . The overall rate of sterility in the end of processing is 94% and for each group this is: 95.4% for MOD, 96.8% for RHT and 86.3% for NBHD . Analysis shows that there is no influence of time of exposure in AB in the rate of decontamination for MOD and RHT . The most predominant germ in contamination is Coagulase Negative Staphylococcus (CNS) (53.4% alone, 8.9% with other bacteria) . 83.3% of MOD; 88.5% of RHT were contaminated with one germ, while 40.4% of NBHD were contaminated with more than one. J Ethnopharmacol, 2005 Jan 4, 96(1-2), 177 - 81 Antimicrobial activity of some medicinal plants of the island Soqotra; Mothana RA et al.; Twenty-five selected plants belonging to 19 families were collected from different localities of the island Soqotra, dried and extracted with the solvents chloroform, methanol and hot water to yield 80 extracts . The extracts were tested for their antimicrobial activity against several Gram-positive and Gram-negative bacteria and against one yeast species using agar diffusion method . Antibacterial activity was demonstrated especially against Gram-positive bacteria including multiresistant Staphylococcus strains . The greatest activity was exhibited by the methanolic extracts of Boswellia elongata, Boswellia ameero, Buxus hildebrandtii, Commiphora parvifolia, Jatropha unicostata, Kalanchoe farinacea, Pulicaria stephanocarpa, Punica protopunica, Withania adunensis and Withania riebeckii . Only the methanolic extract of Buxus hildebrandtii displayed significant antifungal activity. Asian Cardiovasc Thorac Ann, 2004 Dec, 12(4), 316 - 9 Surgery for active, culture negative endocarditis; Hasegawa MI et al.; Culture negative infective endocarditis (CNE) poses very difficult problems during treatment . In this study it was found that of 132 surgically treated patients with infective endocarditis, causative organism was not identified in 46 (34.8 %) . Pre- and perioperative conditions and clinical results of these patients were evaluated . CNE remained very frequent even in these years, and it did not decrease with time . Antibiotic treatment prior to microbiological examinations was commonly observed (nearly 90% orally, and 70% intravenously) . In average, it took more than 2 months to establish the diagnosis of CNE after the onset, and both aortic and mitral valves were affected frequently (19.0 %) . New York Heart Association functional class IV was observed significantly more commonly (61.9%) than culture positive patients . Frequencies of prosthetic valve endocarditis (12.2%), periannular abscess (36.3%), and embolism (21.4%) were similar . Infection was fairly controllable before surgery in 43.9% of CNE patients and in-hospital mortality rate was 14.3%, both of which were comparable to those of all culture positive patients . However, recurrence rate was relatively higher (10.0%) . The conditions and outcomes of CNE were comparable to Staphylococcal endocarditis in some aspects, and were relatively worse than overall culture positive endocarditis. Vet Dermatol, 2004 Dec, 15(6), 369 - 76 A prospective study of the clinical findings, treatment and histopathology of 44 cases of pyotraumatic dermatitis; Holm BR et al.; Pyotraumatic dermatitis (hot spot) is a common clinical syndrome in dogs but there are few prospective scientific studies related to it . The aim of this study was to investigate correlations among clinical pyotraumatic dermatitis, histopathology of the lesions and possible predisposing causes . The relationship of these with breed, age, sex and location of lesion was assessed statistically . A clinical diagnosis of acute pyotraumatic dermatitis was made in 44 privately owned dogs . Males exceeded females (P = 0.0348) and lesions were more common in dogs aged 4 years or less (P < 0.0001) . Lesions were most often seen on the cheek, neck and lateral thigh with a significant correlation between breed and site of lesion (P < 0.0001) . In 31 cases a possible underlying cause was found or suspected . In contrast to previous studies, no otitis externa was recorded and the study was conducted in an area without endemic fleas . Fourteen breeds were represented of which Rottweiler, German shepherd dog and golden retriever were most common . There was no significant seasonal incidence and no correlation among site of lesion and cause, time of year, age or sex . Histopathologically, the dogs could be separated into four patterns by the presence or absence of eosinophils and/or folliculitis . Eosinophils have not previously been recorded in pyotraumatic dermatitis but were seen in 29 cases . Acute folliculitis was seen in 20 cases . However, no correlation was seen among age, sex, breed, underlying cause or site of lesion and histopathology . Twenty-seven cases were cultured for bacteria of which 25 grew Staphylococcus intermedius and two were negative. J Leukoc Biol . 2004 Dec 6; {Epub ahead of print} Mammalian defensins: structures and mechanism of antibiotic activity; Sahl HG et al.; Antibiotic peptides are important effector molecules in host-parasite interactions throughout the living world . In vertebrates, they function in first-line host defense by antagonizing a wide range of microbes including bacteria, fungi, and enveloped viruses . The antibiotic activity is thought to be based on their cationic, amphipathic nature, which enables the peptides to impair vital membrane functions . Molecular details for such activities have been elaborated with model membranes; however, there is increasing evidence that these models may not reflect the complex processes involved in the killing of microbes . For example, the overall killing activity of the bacterial peptide antibiotic nisin is composed of independent activities such as the formation of target-mediated pores, inhibition of cell-wall biosynthesis, formation of nontargeted pores, and induction of autolysis . We studied the molecular modes of action of human defense peptides and tried to determine whether they impair membrane functions primarily and whether additional antibiotic activities may be found . We compared killing kinetics, solute efflux kinetics, membrane-depolarization assays, and macromolecular biosynthesis assays and used several strains of Gram-positive cocci as test strains . We found that membrane depolarization contributes to rapid killing of a significant fraction of target cells within a bacterial culture . However, substantial subpopulations appear to survive the primary effects on the membrane . Depending on individual strains and species and peptide concentrations, such subpopulations may resume growth or be killed through additional activities of the peptides . Such activities can include the activation of cell-wall lytic enzymes, which appears of particular importance for killing of staphylococcal strains. Int J Food Microbiol, 2005 Jan 1, 97(3), 277 - 84 Ecology and characterization by molecular methods of Staphylococcus species isolated from fresh sausages; Rantsiou K et al.; Staphylococcus spp . ecology in fresh sausages stored at 4 degrees C for a period of 10 days was investigated . Microbiological analyses to assess the quality and safety of the products studied were carried out, and strains on MSA agar were isolated . A total of 85 strains were identified, by traditional methods, as Staphylococcus spp . and a PCR-DGGE method, together with a S . xylosus-specific PCR, were used to determine the species of the strains isolated . Almost 50% of them were recognized as S . xylosus, but strains of S . pasteuri, S . warneri, S . equorum and S . succinus, were also found . Interesting population dynamics were observed, characterized by a succession of S . pasteuri and S . warneri at 0 day, with S . xylosus at 3 days and S . equorum at 6 and 10 days . Molecular characterization of S . xylosus strains and cluster analysis highlighted the presence of six main populations in the fresh sausages studied . However, the clusters were formed by strains isolated at different days of storage, implying a homogeneous distribution of the six subpopulations throughout the period in the products followed. J Pharm Pharm Sci, 2004 Nov 17, 7(3), 359 - 71 Influence of the pro-inflammatory cytokines on P-glycoprotein expression and functionality; Fernandez C et al.; PURPOSE: P-glycoprotein (P-gp) is involved in the transport of many drugs at different barriers with consequence in terms of drug distribution and elimination . The expression and activity of P-gp can be modulated by different factors and pathologies . The present article reviews the knowledge regarding the effect of pro-inflammatory cytokines (TNFalpha, IL-1beta, IL-6, IL-2, IFNgamma) on the expression and the functionality of P-gp at three major sites of drug absorption and disposition: the liver, the blood-brain barrier, and the intestine . METHODS: The various methods used to study the effect of pro-inflammatory cytokines include in vivo models (i.e . animals infected with Staphylococcus sp, animals injected with bacterial lipopolysaccharide or directly with cytokines, ...) and in vitro models (i.e . primary rat hepatocytes, human brain endothelial cells, ...) . RESULTS: The data on P-gp expression and/or function may differ according to the compound used to induce inflammation . However, there is a general trend towards a decrease in both the expression of P-gp (mRNA and protein) and its functionality . Transcription factors and nuclear receptors are probably involved in this regulation . CONCLUSION: Cytokines may interfere with P-gp . Hence, in pathological conditions (inflammation, infection, ...), the expression and functionality of P-glycoprotein may be modulated with consequences for drug disposition and, consequently treatment efficacy. Pediatr Dermatol, 2004 Nov-Dec, 21(6), 667 - 9 Generalized bullous impetigo in a neonate; Sandhu K et al.; Generalized bullous impetigo is uncommon in healthy and term neonates . It must be differentiated from staphylococcal scalded skin syndrome and other commonly encountered vesiculopustular lesions . We report generalized bullous impetigo in an otherwise healthy neonate. Nephrol Dial Transplant, 2004 Dec, 19(12), 3155 - 9 Acute glomerulonephritis occurring during immunoadsorption with staphylococcal protein A column (Prosorba); Iglesias J et al.; BACKGROUND: Apheresis of patient plasma by immunoadsorption with a staphylococcal protein A (SPA) column is used in a variety of autoimmune disorders . Leukocytoclastic vasculitis is an uncommon severe complication that can occur during immunoadsorption with SPA (Prosorba . METHODS: We report a case of immune complex glomerulonephritis occurring during Prosorba immunoabsorption in a patient with rheumatoid arthritis (RA) . Using a Medline literature search and information provided by Cypress Bioscience/Fresenius Hemocare, we review renal complications associated with Prosorba immunoadsorption . RESULTS: We identified seven additional potential cases of glomerulonephritis (GN) in association with Prosorba immunoadsorption . Five of these patients were being treated for RA, and two for idiopathic thrombocytopenia purpura (ITP) . Renal biopsies were performed on four patients, all of whom had evidence of immune complex GN . Among RA patients treated with Prosorba, the incidence of GN closely paralleled that of leukocytoclastic vasculitis at 1.75% . The presence of leukocytoclastic vasculitis was a significant risk factor for the development of GN (relative risk = 75.95, CI 7-1869, P = 0.00021) . In contrast, among more than 10 000 ITP patients treated with Prosorba, there were only two potential cases of GN . The risk of developing GN in association with Prosorba immunoadsorption was significantly greater for patients with RA than for those with ITP (relative risk = 62.95, CI 10-453, P = 0.00002) . CONCLUSION: This case series highlights the risk of GN among patients undergoing SPA immunoadsorption . The development of GN is associated with the presence of leukocytoclastic vasculitis . Patients with RA seem to be at particular risk. Proteins, 2005 Feb 1, 58(2), 271 - 7 Elucidation of information encoded in tryptophan 140 of staphylococcal nuclease; Hirano S et al.; We investigated the role of W140 in the folding of Staphylococcal nuclease . For this purpose, we constructed the 19 possible substitution mutations at residue 140 . Only three mutants, W140F, W140H, and W140Y, adopted native-like structures under physiological conditions and showed native-like enzymatic activities . In contrast, the other 16 mutants took on compact unfolded structures under physiological conditions and the enzymatic activities of these mutants were decreased to approximately 70% of wild-type levels . These 16 mutants maintained substrate-induced foldability . These results strongly indicate that the side-chain information encoded by residue 140 is essential to maintain a stable native structure, and that this residue must be an aromatic side chain . The order of thermal stability was wild type > W140H > W140F = W140Y . Therefore, the five-membered nitrogen-containing ring of the indole is thought to bear the essential information . In the crystal structure of staphylococcal nuclease, the five-membered ring is at the local center of the C-terminal cluster through hydrophobic interactions . This cluster plays a key role in the interaction connecting the C-terminal region and the N-terminal beta-core . Mutants other than W140H, W140F, and W140Y lost the ability to form the local core, which caused the loss of the long-range interactions between the C-terminal and N-terminal regions . Inhibitor or substrate binding to these mutants compensates for the lack of long-range interactions generated by W140 . (c) 2004 Wiley-Liss, Inc. J Autoimmun, 2004 Dec, 23(4), 379 - 83 IL-10 production in B cells is confined to CD154+ cells in patients with systemic lupus erythematosus; Diaz-Alderete A et al.; The immunological hallmark of SLE is B cell hyperactivity . CD154 (CD40-L) is normally expressed in activated T cells, and plays an important role in T-B interactions . Its expression is increased in SLE T cells . Additionally, its expression on B cells leads to the development of SLE-like disease in a transgenic model . IL-10 is a key cytokine in the disturbed SLE immune system . The aim of this work was to explore the relation between IL-10 and CD154 expression in SLE B cells . We studied 11 SLE patients and 10 healthy volunteers . Mononuclear cells were isolated from peripheral blood and cultured in the presence or absence of Cowan I Strain Staphylococcus (CSS) . Surface CD154 and intracytoplasmic IL-10 expression were quantified with flow cytometry . In basal conditions, CD154 expression was not different in patients and controls . B cell stimulation did not cause a significant increase in CD154 expression in control B cells . However, its expression increased 2 times in B cells obtained from SLE patients . IL-10 expression was confined to CD154(+) cells . Our results show that IL-10 production is intimately linked to CD154 expression in B cells, and that the IL-10(+)CD154(+) B cell subset increases abnormally when SLE-derived cells are stimulated with CSS. J Hosp Infect, 2005 Jan, 59(1), 19 - 26 Genomic characterization of oxacillin-resistant Staphylococcus epidermidis and Staphylococcus haemolyticus isolated from Brazilian medical centres; Nunes AP et al.; Studies on the genetic diversity of oxacillin-resistant coagulase-negative staphylococcal (CNS) isolates are important for the control and prevention of infections . The present study evaluated the clonal diversity of oxacillin-resistant Staphylococcus epidermidis (ORSE) and Staphylococcus haemolyticus (ORSH) strains, isolated from patients in nine Brazilian medical centres by using pulsed-field gel electrophoresis (PFGE) after digestion of bacterial DNA using SmaI . PFGE analysis of ORSE (N=44) and ORSH (N=25) strains showed the presence of 29 restriction profiles clustered in 16 PFGE types, and 21 distinct profiles in 15 PFGE types, respectively, indicating a large genetic diversity among isolates of both of these species . Among the ORSE isolates, 23 (52%) strains belonged to two predominant PFGE types (named A and B), which were observed in most of the hospitals assessed, indicating the spread of these PFGE types in hospitals located in Rio de Janeiro . The spread of PFGE types of ORSH was also detected in some of the hospitals investigated . The results show that PFGE is a suitable tool for epidemiological studies of oxacillin-resistant CNS, and can be used as a basis for infection control procedures for these multiresistant organisms. Ann Surg, 2004 Dec, 240(6), 1065 - 72; discussion 1072-3 Staphylococcal enterotoxin A-induced hepatotoxicity is predominantly mediated by Fas ligand (CD95L); Klintman D et al.; OBJECTIVE: To determine the role of tumor necrosis factor alpha (TNF-alpha) and Fas ligand (FasL, CD95L) in superantigen-induced and endotoxin-induced liver injury . SUMMARY BACKGROUND DATA: Gram-positive bacteria are increasingly common causes of sepsis and multiorgan failure, but the pathophysiologic mechanisms of superantigen-provoked hepatotoxicity remain elusive . METHODS: Intravital fluorescence microscopy was used to study the liver microcirculation in mice challenged with superantigen (staphylococcal enterotoxin A, SEA) or endotoxin (lipopolysaccharide, LPS) combined with D-galactosamine . RESULTS: Administration of 10 microg LPS and 50 microg SEA caused similar hepatocellular damage as determined by liver enzymes and apoptosis . Notably, TNF-alpha-deficient mice were completely protected against hepatic injury provoked by LPS, whereas no protection was observed in response to SEA . On the other hand, FasL-deficient mice were protected against liver injury induced by SEA, but no protection was found when challenged with LPS . LPS increased clear-cut leukocyte recruitment, whereas SEA had no significant effect on leukocyte responses in the liver microcirculation . Leukocyte responses to LPS were decreased by >56% in TNF-alpha gene-targeted animals . Moreover, antiadhesive therapy, ie, immunoneutralization of P-selectin, which is an effective inhibitor of leukocyte recruitment, protected against LPS-induced but not against SEA-induced hepatic damage . CONCLUSIONS: These novel findings demonstrate that the mechanisms of hepatic injury in endotoxin-induced and superantigen-induced sepsis are principally different . On one hand, SEA-provoked hepatotoxicity is mediated by FasL and is not associated with leukocyte recruitment . On the other hand, liver damage provoked by LPS is mediated by TNF-alpha and characterized by prominent leukocyte responses . These data may facilitate development of more specific therapies against sepsis of different origins. J Chemother, 2004 Oct, 16(5), 446 - 52 Polyurethanes loaded with antibiotics: influence of polymer-antibiotic interactions on in vitro activity against Staphylococcus epidermidis; Piozzi A et al.; Acidic or basic polyurethanes were loaded with antibiotics to develop materials to prevent medical device-related infections . A correlation between polymer-antibiotic interactions and amount of drug absorbed by polymers and released over time was found . Since the employed antibiotics, i.e . amoxicillin, cefamandole nafate, rifampin and vancomycin, possessed at least an acidic group in their structural formula, the introduction of basic tertiary amines in the polyurethane side-chain resulted in an increased polymer ability to adsorb antibiotics . However, a stronger ionic interaction between this polymer and the antibiotics caused a release of lower amount of drug over time . Antibiotics released from polymers inhibited Staphylococcus epidermidis growth on agar . Antibiotic-loaded polyurethanes kept in water for increasing times were still able to show inhibition zones of bacterial growth . The antibacterial activity lasted up to 3 hours for amoxicillin, 24 hours for vancomycin, 8 days for cefamandole nafate and 8 months for rifampin. J Contemp Dent Pract, 2004 Nov 15, 5(4), 91 - 100 Qualitative and quantitative analysis of bacterial aerosols; Al Maghlouth A et al.; The objective of this study was to investigate qualitatively and quantitatively the bacterial aerosols before, during, and after clinical work sessions in different areas within a multichair dental clinic, an isolation clinic, the sterilization center, and the prosthetic laboratory in the College of Dentistry, King Saud University . Also, the contributions of aerosols generated by different types of dental procedures were investigated . Air sampling using blood and heart infusion agar plates at four selected areas was performed three times per day over a 2-week period before, during, and after clinical sessions . The concentration of total bacterial aerosols was 5 times higher in the multichair clinic, 3.6 times higher in the prosthetic laboratory, 2 times higher in the sterilization center and isolation clinic during working sessions as compared to before the working sessions . At the end of the working day, aerosols decreased 50-70% in all areas . Staphylococcus epidermidis had the highest prevalence (37.12q%) of colony composition of bacteria examined . This study demonstrates that aerosols increase during and after work sessions and, therefore, increase the chance for infectious agent transmission . Preventive measures should be instituted to reduce or disrupt aerosols as a transmission route in the multichair dental clinic, sterilization center, prosthetic laboratory, and isolation clinic. J Vasc Surg, 2004 Nov, 40(5), 899 - 906 Total laparoscopic bypass for aortoiliac occlusive lesions: 93-case experience; Coggia M et al.; OBJECTIVES: We describe our experience with a new technique of total laparoscopic bypass surgery to treat aortoiliac occlusive lesions . MATERIAL AND METHODS: From November 2000 to December 2003, 93 total laparoscopic bypass procedures were performed to treat TASC (TransAtlantic Inter-Society Consensus document) grade C or D aortoiliac occlusive lesions . We also reimplanted 2 inferior mesenteric arteries, and performed 3 prosthesis-superior mesenteric bypasses and 2 suprarenal aorta endarterectomies . Our technique includes a sloping right lateral decubitus installation, which enables a simple transperitoneal left retrocolic or retrorenal approach to the infrarenal abdominal aorta . In patients with a hostile abdomen a retroperitoneal videoscopic approach was used . Aorta-prosthesis laparoscopic anastomoses are performed simply, which averts any trauma to the suture material . RESULTS: Patients included 76 men and 17 women, with median patient age 61 years (range, 38-79 years) . The approach to the aorta was always possible, in particular, in obese patients . It enabled stable aortic exposure during performance of the laparoscopic aorta-prosthesis anastomosis . Median operative time was 240 minutes (range, 150-450 minutes) . Median aortic clamping time measured to unclamping of the first prosthetic limb was 67.5 minutes (range, 30-135 minutes) . Median duration of aorta-prosthesis anastomosis was 30 minutes (range, 12-90 minutes) . The longest durations were mainly observed during the learning curve . Thirty-day postoperative mortality was 4% (4 of 93 patients) . Two patients died of myocardial infarction . One patient with American Society of Anesthesiologists grade 4 disease operated on to treat critical ischemia died of multiple organ system failure, and 1 patient died of colonic ischemia . Major nonlethal postoperative complications were observed in 4 patients, and included lung atelectasia in 2 patients, graft infection in 1 patient operated on emergently to treat aortic occlusion, and secondary spleen rupture at day 5 in 1 patient . Median hospital stay was 7 days (range, 2-57 days) . With a mean follow-up of 19 months (range, 1-37 months), complete recovery was observed in 89 patients, and all grafts were patent . One patient had kinking of a prosthetic limb at the groin, and in 1 patient Staphylococcus epidermidis graft infection developed, which was treated with in situ replacement with a rifampin-bonded graft . CONCLUSION: Total laparoscopic aortic bypass is feasible . In patients with TASC C and D aortoiliac occlusive lesions, short-term outcomes are comparable to those with conventional aortic bypass . After the initial learning curve, laparoscopic technique may reduce the operative trauma of aortic bypass. Protein Sci, 2004 Dec, 13(12), 3104 - 14 Infrequent cavity-forming fluctuations in HPr from Staphylococcus carnosus revealed by pressure- and temperature-dependent tyrosine ring flips; Hattori M et al.; Infrequent structural fluctuations of a globular protein is seldom detected and studied in detail . One tyrosine ring of HPr from Staphylococcus carnosus, an 88-residue phosphocarrier protein with no disulfide bonds, undergoes a very slow ring flip, the pressure and temperature dependence of which is studied in detail using the on-line cell high-pressure nuclear magnetic resonance technique in the pressure range from 3 MPa to 200 MPa and in the temperature range from 257 K to 313 K . The ring of Tyr6 is buried sandwiched between a beta-sheet and alpha-helices (the water-accessible area is less than 0.26 nm2), its hydroxyl proton being involved in an internal hydrogen bond . The ring flip rates 10(1)-10(5) s(-1) were determined from the line shape analysis of H(delta1, delta2) and H(epsilon1,epsilon2) of Tyr6, giving an activation volume DeltaV++ of 0.044 +/- 0.008 nm3 (27 mL mol(-1)), an activation enthalpy DeltaH++ of 89 +/- 10 kJ mol(-1), and an activation entropy DeltaS++ of 16 +/- 2 JK(-1) mol(-1) . The DeltaV++) and DeltaH++ values for HPr found previously for Tyr and Phe ring flips of BPTI and cytochrome c fall within the range of DeltaV(double dagger) of 28 to 51 mL mol(-1) and DeltaH++ of 71 to 155 kJ mol(-1) . The fairly common DeltaV++ and DeltaH++ values are considered to represent the extra space or cavity required for the ring flip and the extra energy required to create a cavity, respectively, in the core part of a globular protein . Nearly complete cold denaturation was found to take place at 200 MPa and 257 K independently from the ring reorientation process. Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi, 2004 Nov, 20(6), 757 - 9 {Study on the TCR Vbeta binding sites in the superantigen staphylococcal enterotoxin D.}; Li YF et al.; AIM:To study TCR Vbeta binding sites of staphylococcal enterotoxin D(SED) . METHODS: Six SED mutants were constructed by site-directed mutagenesis . The activity of promoting T cell proliferation by the mutants was detected by (3)H-TdR incorporation . For the mutants with decreased mitogenic activity, flow cytometry was used detect their MHC-II binding activity and TCR Vbeta specificity . RESULTS: Residue N23 played an important role in the interaction of SED with human TCR Vbeta5 . Residue H26 was probably a SED binding site to human TCR Vbetas except for TCR Vbeta5, TCR Vbeta8 and TCR Vbeta12.1 . CONCLUSION: Residue N23 is a key TCR Vbeta binding site of SED. Ann Ig, 2004 Jan-Apr, 16(1-2), 247 - 53 {Mycobacteria in swimming pool water and the meaning of microbiological conventional indicators}; Reali D et al.; Monitoring program of hygienic quality water in twelve public swimming pools was performed . Legally required microbiological indicator parameters of safety for gastrointestinal illness were measured besides the analyses of Pseudomonas spp . and Staphylococcus spp . prevalence, frequency of recovery and number of nontuberculous mycobacteria . We detected positive samples for coliforms at lower rate (29.3%) than Pseudomonas (75.5%), Staphylococcus spp . (46%) and Mycobacteria (59.4%) . We pointed out statistically significant correlation (r=0.67 p=0.0001) between Mycobacteria and Pseudomonas so we think that the latter might be a good predictive marker . As 82% of samples had free chlorine residual within the limits stated by Italian Laws, the efficacy of chlorination to prevent risk of infectious diseases transmission by route other than gastroenteric was discussed . A revision of both the sanitary significance of conventional microbial parameters and the related regulations appears necessary. J Invasive Cardiol, 2004 Nov, 16(11), 632 - 4 Bacteremia following complex percutaneous coronary intervention; Ramsdale DR et al.; BACKGROUND AND OBJECTIVE: Complex percutaneous coronary intervention (PCI) often requires introduction of numerous devices into and out of the arterial circulation and this may result in an increased risk of bacteraemia or even septicaemia . This study was undertaken to detect the frequency of bacteraemia that may be associated with such procedures . METHODS: 147 patients undergoing complex PCI had blood culture tests immediately after and 12 hours after the procedure . RESULTS: Of 147 patients, 26 (17.7%) had detectable bacteraemia immediately after PCI . Coagulase-negative staphylococcus was isolated most commonly . An additional 12% of patients yielded positive blood cultures in the next 12 hours with femoral sheaths still in-situ . There were no associated clinical sequelae . CONCLUSIONS: Uncomplicated bacteraemia is not uncommon as a result of complex PCI procedures . Although there are usually no clinical sequelae, these findings are important for those patients who are considered to be at moderate or high risk of infective endocarditis who require an invasive procedure such as PCI . This paper emphasizes the need for maximum sterility during PCI procedures if infective complications and stent infection are to be avoided. Physiol Genomics, 2004 Nov 17, 19(3), 247 - 54 Whole blood and leukocyte RNA isolation for gene expression analyses; Feezor RJ et al.; The analysis of gene expression data in clinical medicine has been plagued by the lack of a critical evaluation of accepted methodologies for the collection, processing, and labeling of RNA . In the present report, the reliability of two commonly used techniques to isolate RNA from whole blood or its leukocyte compartment was compared by examining their reproducibility, variance, and signal-to-noise ratios . Whole blood was obtained from healthy subjects and was either untreated or stimulated ex vivo with Staphylococcus enterotoxin B (SEB) . Blood samples were also obtained from trauma patients but were not stimulated with SEB ex vivo . Total RNA was isolated from whole blood with the PAXgene proprietary blood collection system or from isolated leukocytes . Biotin-labeled cRNA was hybridized to Affymetrix GeneChips . The Pearson correlation coefficient for gene expression measurements in replicates from healthy subjects with both techniques was excellent, exceeding 0.985 . Unsupervised analyses, including hierarchical cluster analysis, however, revealed that the RNA isolation method resulted in greater differences in gene expression than stimulation with SEB or among different trauma patients . The intraclass correlation, a measure of signal-to-noise ratio, of the difference between SEB-stimulated and unstimulated blood from healthy subjects was significantly higher in leukocyte-derived samples than in whole blood: 0.75 vs . 0.46 (P = 0.002) . At the P < 0.001 level of significance, twice as many probe sets discriminated between SEB-stimulated and unstimulated blood with leukocyte isolation than with PAXgene . The findings suggest that the method of RNA isolation from whole blood is a critical variable in the design of clinical studies using microarray analyses. Lett Appl Microbiol, 2004, 39(6), 490 - 4 Comparison of three immunological methods for detecting staphylococcal enterotoxins from food; Vernozy-Rozand C et al.; AIMS: Immunologically based assays for the detection of staphylococcal enterotoxins are numerous . These techniques include radio immunosorbent assays and enzyme-linked immunosorbent assays (ELISA), some of which are available as commercial kits . The purpose of this study was to compare the performances of three commercial immunoassays . METHODS AND RESULTS: Two automated detection systems, VIDAS SET bioMerieux, VIDAS SET2 bioMerieux and an ELISA method, TRANSIA PLATE Staphylococcal Enterotoxins Diffchamb were compared for detecting different quantities of purified staphylococcal enterotoxins (A, B, C2, D and E) added to food . CONCLUSIONS: VIDAS SET2 had a greater specificity (100%) and sensitivity than VIDAS SET and TRANSIA PLATE Staphylococcal Enterotoxins . More precisely, VIDAS SET2 could detect <0.5 ng g(-1) of toxins A and B, <1 ng g(-1) of toxins C2 and E and 1 ng g(-1) of toxins D and E . SIGNIFICANCE AND IMPACT OF THE STUDY: Because staphylococcal food poisoning (resulting from ingestion of low levels of staphylococcal enterotoxins) is one of the most common forms of foodborne illness there is a need for specific and sensitive methods for detecting these enterotoxins . VIDAS SET2 appears to be suitable for detecting staphylococcal enterotoxins from food. Angiology, 2004 Nov-Dec, 55(6), 697 - 9 Bacteremia caused by late-infected pacemaker lead--a case report; Kinoshita O et al.; A 73-year-old man with bradycardia and atrial fibrillation underwent implantation of a transvenous pacemaker system on the left anterior chest wall in 1995 . Six years later, he was admitted for bacteremia from coagulase-negative Staphylococcus . Repeated treatment employing antibiotic therapy was ineffective . The infected electrode was removed under cardiopulmonary bypass . His electrode had become firmly encased with fibrous tissue within the right ventricle and atrium . It was removed under direct vision during complete cardiac arrest . The postoperative course was uneventful and there has been no recurrence after 1 year. J Clin Invest, 2004 Nov, 114(10), 1484 - 92 Defining the pathogenic involvement of desmoglein 4 in pemphigus and staphylococcal scalded skin syndrome; Nagasaka T et al.; Desmogleins (Dsgs), cadherin-type cell adhesion molecules, are targeted in skin-blistering diseases such as pemphigus and staphylococcal scalded skin syndrome (SSSS) . The role of Dsg4, a new isoform, was investigated in these diseases . Dsg4 was recognized by 30 (77%) of 39 pemphigus sera containing anti-Dsg1 IgG but not by 16 pemphigus sera containing no anti-Dsg1 IgG or by 34 normal control sera . The Dsg4 immunoreactivity of these sera was abolished by removal of anti-Dsg1 IgG . Conversely, the removal of anti-Dsg4 IgG from pemphigus sera reduced the immunoreactivity against Dsg1 only 13.8% +/- 8.8% (n = 23) and did not affect its ability to induce blisters in neonatal mice . IgG that was affinity-purified on Dsg4 recognized Dsg1 but failed to induce blisters, while IgG purified on Dsg1 from the same pemphigus foliaceus sera induced blisters . Thus, pemphigus sera show Dsg4 reactivity due to cross-reactivity of a subset of anti-Dsg1 IgG, and the Dsg4/Dsg1-cross-reacting IgG has no demonstrable pathogenic effect . In addition, Dsg4 was not cleaved by exfoliative toxins that induce blisters in SSSS . These findings suggest that Dsg4 may play a role other than adhesion and that the cross-reactivity of desmoglein autoantibodies should be factored into the framework of future studies of autoimmune mechanisms in pemphigus. Int J Syst Evol Microbiol, 2004 Nov, 54(Pt 6), 2213 - 5 Reclassification of Staphylococcus pulvereri Zakrzewska-Czerwinska et al . 1995 as a later synonym of Staphylococcus vitulinus Webster et al . 1994; Svec P et al.; A polyphasic taxonomic approach was applied to strains of the species Staphylococcus vitulinus and Staphylococcus pulvereri in order to clarify their taxonomic relatedness . Four reference strains, representing both species, and seven strains isolated from human clinical material were characterized by biotyping, ribotyping and SDS-PAGE analysis of whole-cell proteins, and none of the screening approaches allowed the two taxa to be distinguished . DNA-DNA hybridization experiments between four selected representative strains, including the type strains, confirmed that Staphylococcus pulvereri is a later synonym of Staphylococcus vitulinus. Neuroscience, 2004, 129(3), 555 - 62 Behavioural endocrine immune-conditioned response is induced by taste and superantigen pairing; Pacheco-Lopez G et al.; Administration of bacterial superantigen, such as staphylococcal enterotoxin B (SEB), induces in vivo stimulation of T cell proliferation and cytokine production such as interleukin-2 (IL-2) . It has been previously reported that SEB administration induces fever, c-Fos expression in the brain, and hypothalamus-pituitary-adrenal axis activation, demonstrating that the brain is able to sense and respond to SEB . Previously it had been shown that immune functions can be behaviourally conditioned pairing a novel gustatory stimulus together with an immunomodulatory drug or an antigen . We designed an experimental protocol using Dark Agouti rats in which saccharin taste, as conditioned stimulus, was paired with an i.p . injection of SEB (2 mg/kg), as unconditioned stimulus . Six days later, when conditioned animals were re-exposed to the conditioned stimulus they displayed strong conditioned taste avoidance to the saccharin . More importantly, re-exposure to the conditioned stimulus significantly increased IL-2, interferon-gamma and corticosterone plasma levels, in comparison with conditioned animals which had not been re-exposed to saccharin taste . These results demonstrate a behavioural-immune-endocrine conditioned response using a superantigen as unconditioned stimulus . In addition, they illustrate the brain abilities to mimic the unconditioned effects of a superantigen by yet unknown mechanisms. Tenn Med, 2004 Oct, 97(10), 453 - 6 Monitoring quality of flexible endoscope disinfection by microbiologic surveillance cultures; Tunuguntla A et al.; BACKGROUND: The purpose of this study is to assess the adequacy of currently approved high-level disinfection (HLD) for flexible endoscopes, by reviewing retrospectively the results of environmental cultures on the endoscopes, which were ready to use . METHODS: Cultures of all of our gastrointestinal endoscopes that were ready for use were sent to the laboratory for environmental cultures . Thirty milliliters of sterile water was passed through the biopsy channel of the endoscopes and the flushed water was collected in a sterile container . The specimen was plated onto blood agar, incubated at 37C, and examined for growth at 24 and 48 hours . RESULTS: Cultures were positive during the initial part of the study repeatedly with the same organism; the source of contamination was traced to be the source water and scope cleaner, after they were suspected and sent for cultures . In addition to Pseudomonas, there was contamination of the endoscopes with Micrococcus and Staphylococcus during the study period . CONCLUSIONS: Using environmental cultures for monitoring effectiveness of standard reprocessing procedures on a regular basis in clinical practice is not only inexpensive but also valuable to identify the source of possible contamination and rectify reprocessing methods to prevent transmission of infection secondary to endoscopy. J Allergy Clin Immunol, 2004 Nov, 114(5), 1059 - 69 Superantigen-induced corticosteroid resistance of human T cells occurs through activation of the mitogen-activated protein kinase kinase/extracellular signal-regulated kinase (MEK-ERK) pathway; Li LB et al.; BACKGROUND: Microbial superantigens induce human T-cell resistance to corticosteroids . OBJECTIVE: Understanding the molecular pathways resulting in corticosteroid-resistant T cells is important because this condition can complicate the treatment of inflammation . METHODS: The response of human PBMCs to steroids was assessed by using proliferation assays after stimulation with superantigens or anti-CD3 in the presence of various kinase inhibitors . Glucocorticoid receptor alpha (GCRalpha) localization was defined on the basis of intracellular staining . Protein phosphorylation was measured by means of Western blotting . RESULTS: In the current study we found that PBMCs stimulated with superantigen, but not anti-CD3, induced corticosteroid-resistant T cells . However, the purified T cells stimulated either with staphylococcal enterotoxin B (SEB) or anti-CD3 are susceptible to corticosteroid inhibition . These results imply that signals on antigen-presenting cells might act in concert with the T-cell receptor to cause steroid resistance . Blockade of CD40-CD40 ligand interaction had no effect on superantigen-induced corticosteroid resistance . However, CD28 costimulation with T-cell receptor activation induced corticosteroid resistance of human T cells in a dose-dependent manner . Superantigen stimulation, compared with anti-CD3 stimulation, was found to induce a more rapid and sustained phosphorylation of mitogen-activated extracellular signal-regulated kinase (ERK) . Treatment with PD98059 and UO126 (specific mitogen-activated protein kinase kinase {MEK}/ERK inhibitors), but not a p38 inhibitor or a c-Jun N-terminal kinase inhibitor, restored the response to steroids, as indicated by proliferation assays . Furthermore, purified ERK1 and ERK2 were able to phosphorylate recombinant human GCRalpha directly in an in vitro kinase assay . Of note, superantigen-induced corticosteroid resistance was associated with abrogation of GCRalpha nuclear translocation . This effect could be reversed by treatment with MEK/ERK pathway inhibitors . CONCLUSIONS: These data are compatible with the hypothesis that superantigen-induced corticosteroid resistance involves the Raf-MEK-ERK1/ERK2 pathway of T-cell receptor signaling, which leads to GCRalpha phosphorylation and inhibition of dexamethasone-induced GCRalpha nuclear translocation. Anaesth Intensive Care, 2004 Oct, 32(5), 707 - 10 Successful use of ECMO in adults with life-threatening infections; MacLaren G et al.; Two cases of critically ill patients who received extracorporeal membrane oxygenation (ECMO) using different forms of circuitry and for different indications are presented . Both patients had life-threatening infections with septic shock and were not able to be supported by conventional means . The first patient had staphylococcal septicaemia and received venoarterial ECMO for circulatory failure . The second patient had psittacosis and received venovenous ECMO for respiratory failure . We discuss the expanding indications for this technology and the role it has to play in adult intensive care. Folia Neuropathol, 2004, 42(3), 125 - 32 Toll-like receptors in rat brains injured by hypoxic-ischaemia or exposed to staphylococcal alpha-toxin; Maslinska D et al.; Some data suggest that the central nervous system (CNS) is the main target of Staphylococcus alpha-toxin . Since this pathogen cannot penetrate the blood-brain barrier (BBB), the exact mechanism by which alpha-toxin affects the CNS remains unclear . Recent studies on the role of the innate immune system have shed light on how bacterial infections initiate inflammatory responses within the CNS . The aim of this study was to investigate the immunoexpression of Toll-like receptors (TLR 2, TLR 4) in brains of young rats systemically exposed to Staphylococcus alpha-toxin or injured by neonatal hypoxia-ischaemia . The study was carried out on 6-week-old Wistar rats . A group of 6-week-old rats with severe brain injury caused by neonatal hypoxia-ischaemia was also studied separately . In all control rats, the immunoexpression of TLR 2 and TLR 4 was not detected . However, the expression of both TLRs was evident in all brains injured by HI or exposed to alpha-toxin . The immunoexpression was localised in the wall of the small brain vessels, cells of ependyma and leptomeninges . In such vessels the spectrum of ultrastructural lesions was found . The presence of TLR4 detected in the nerve cells of the subcortical gray matter of the brain is particularly of interest, but requires further studies . The presence of TLR 4 antigen in the nerve cells of the subcortical gray matter is particularly of interest . In conclusion, the results show that brain microvessels through TLRs may participate in the immune response of brain affected by bacterial infection as well as injured by non-infection insults. Mol Cell Biochem, 2004 Jul, 262(1-2), 177 - 85 Inhibition of nuclear factor-kappa B prevents staphylococcal enterotoxin A-induced fever; Shao DZ et al.; It has been shown that staphylococcal enterotoxin A (SEA) acts through human peripheral blood mononuclear cells (PBMC) to stimulate synthesis or release of pyrogenic cytokines . Nuclear factor-kappa B (NF-kappaB) is thought to play an important role in inflammatory responses through the regulation of genes encoding pro-inflammatory cytokines . The purpose of the present study was to determine whether the NF-kappaB mechanisms in human PBMC are involved in SEA-induced fever . Western blot evaluation revealed SEA was able to induce nuclear translocation of NF-kappaB from cytosol to nucleus in PBMC, which could be abolished by a NF-kappaB inhibitor such as pyrrolidine dithiocarbamate (PDTC), sodium pyrithione (Pyri), N-acetyl-L-cysteine (NAC), or curcumin (Cur) . Electrophoretic mobility shift assay also showed that the NF-kappaB DNA-binding activity was increased in the SEA-treated PBMC . Again, the SEA-induced increased NF-kappaB binding activity was significantly attenuated by either PDTC, Pyri, NAC or Cur . The pyrogenic responses to supernatant fluids obtained from human PBMC stimulated with SEA were associated with increased levels of interleukin 1-beta (IL-1beta), IL-6, and tumor necrosis factor-alpha (TNF-alpha) in the supernatant fluids . Both the fever and the increased levels of IL-1beta, IL-6, and TNF-alpha in supernatant fluids obtained from the SEA-stimulated PBMC were decreased by incubating SEA-PBMC with either PDTC, Pyri, NAC, or Cur . Furthermore, the fever induced by systemic or central administration of SEA in rabbits were attenuated by pre-treatment with an systemic or central dose of either PDTC, Pyri, NAC, or Cur . The data indicate that inhibition of NF-kappaB prevents SEA-induced fever. J Clin Microbiol, 2004 Nov, 42(11), 5324 - 6 Prevalence and characterization of leukotoxin-producing Staphylococcus intermedius in Isolates from dogs and pigeons; Futagawa-Saito K et al.; Staphylococcus intermedius isolates from dogs (n = 44) and pigeons (n = 62) were categorized into 12 types by intergenic ribosomal DNA spacer polymorphism analysis . All isolates from pigeons were lukS positive and all isolates from dogs were lukS and lukF positive by dot blot analysis . The mean leukotoxicity titer for dog isolates was at least 129-fold higher than that for pigeon isolates. Proc Natl Acad Sci U S A, 2004 Nov 16, 101(46), 16168 - 73 Epub 2004 Nov 16. Hydrophobic interactions drive ligand-receptor recognition for activation and inhibition of staphylococcal quorum sensing; Wright JS 3rd et al.; Two-component systems represent the most widely used signaling paradigm in living organisms . Encoding the prototypical two-component system in Gram-positive bacteria, the staphylococcal agr (accessory gene regulator) operon uses a polytopic receptor, AgrC, activated by an autoinducing peptide (AIP), to coordinate quorum sensing with the global synthesis of virulence factors . The agr locus has undergone evolutionary divergence, resulting in the formation of several distinct inter- and intraspecies specificity groups, such that most cross-group AIP-receptor interactions are mutually inhibitory . We have exploited this natural diversity by constructing and analyzing AgrC chimeras generated by exchange of intradomain segments between receptors of different agr groups . Functional chimeras fell into three general classes: receptors with broadened specificity, receptors with tightened specificity, and receptors that lack activation specificity . Testing of these chimeric receptors against a battery of AIP analogs localized the primary ligand recognition site to the receptor distal subdomain and revealed that the AIPs bind primarily to a putative hydrophobic pocket in the receptor . This binding is mediated by a highly conserved hydrophobic patch on the AIPs and is an absolute requirement for interactions in self-activation and cross-inhibition of the receptors . It is suggested that this recognition scheme provides the fundamental basis for agr activation and interference. Am J Infect Control, 2004 Nov, 32(7), 414 - 6 Antimicrobial susceptibility of microorganisms recovered from intraabdominal infections at Belo Horizonte, Brazil; dos Santos SG et al.; We evaluated antimicrobial susceptibility patterns of microorganisms isolated from intraabdominal infection of Brazilian patients, by agar dilution, agar diffusion, and E test . Among the strictly anaerobes, 57.7% were resistant to penicillin, 28.2% to clindamycin, and 9.9% to metronidazole . The majority of Escherichia coli and Staphylococcus were sensitive and resistant to almost all drugs, respectively . Half of Candida samples were resistant to itraconazole . Our data reinforce the importance of this kind of study to support rational antimicrobial therapy. Ginekol Pol, 2004 May, 75(5), 382 - 4 {Staphylococcal toxic shock syndrome}; Malolepsza E et al.; {A case of menstrual toxic shock syndrome (TSS) in a woman aged 21 is reported . Diagnosis was made on clinical basis and was followed by successful treatment} J Biomed Mater Res . 2000 Jun 15;50(4):546-556 {Epub ahead of print} Subcutaneous abscess formation around catheters induced by viable and nonviable Staphylococcus epidermidis as well as by small amounts of bacterial cell wall components; Boelens JJ et al.; The use of catheters is often complicated by infection, mainly due to Staphylococcus epidermidis . Recently, a novel poly(vinylpyrrolidone)-grafted silicone elastomer catheter (SEpvp) was introduced . Less bacteria adhered to SEpvp than to conventional SE catheters in vitro . The frequency of S . epidermidis infection associated with SEpvp and SE was assessed in a rabbit model . Unexpectedly, abscesses were induced by the injection of low numbers of S . epidermidis along subcutaneously inserted SEpvp . No abscesses were seen around SE, even when very high numbers of S . epidermidis were injected . This bioincompatibility reaction observed around the SEpvp was independent of the host, bacterial strain, and method of inoculation . Abscesses were also induced by nonviable S . epidermidis and by bacterial cell wall components . Because these incompatibility reactions were not observed in the absence of bacteria, biocompatibility testing should include experiments in which the inflammatory effects of the combination of catheter and (non)viable bacteria are tested . (c) 2000 John Wiley & Sons, Inc . J Biomed Mater Res, 50, 546-556, 2000. Exp Biol Med (Maywood), 2004 Nov, 229(10), 1061 - 71 Functional piglet model for the clinical syndrome and postmortem findings induced by staphylococcal enterotoxin B; van Gessel YA et al.; Staphylococcal enterotoxin (SE) B causes serious gastrointestinal illness, and intoxication with this exotoxin can lead to lethal toxic shock syndrome . In order to overcome significant shortcomings of current rodent and nonhuman primate models, we developed a piglet model of lethal SEB intoxication . Fourteen-day-old Yorkshire piglets were given intravenous SEB, observed clinically, and sacrificed at 4, 6, 24, 48, 72, or 96 hrs posttreatment . Clinical signs were biphasic with pyrexia, vomiting, and diarrhea within 4 hrs, followed by terminal hypotension and shock by 96 hrs . Mild lymphoid lesions were identified as early as 24 hrs, with severe lymphadenopathy, splenomegaly, and prominent Peyer's patches found by 72 hrs . Widespread edema-most prominent in the mesentery, between loops of spiral colon, and in retroperitoneal connective tissue-was found in animals at 72 hrs . Additional histologic changes included perivascular aggregates of large lymphocytes variably present in the lung and brain, circulating lymphoblasts, and lymphocytic portal hepatitis . Preliminary molecular investigation using gene array has uncovered several gene profile changes that may have implications in the pathophysiology leading to irreversible shock . Five genes were selected for further study, and all showed increased mRNA levels subsequent to SEB exposure . The use of this piglet model will continue to elucidate the pathogenesis of SEB intoxication and facilitate the testing of new therapeutic regimens that may better correlate with human lesions. Biosens Bioelectron, 2004 Nov 1, 20(4), 753 - 64 Fabrication and characterization of 3D hydrogel microarrays to measure antigenicity and antibody functionality for biosensor applications; Charles PT et al.; We report the fabrication, characterization and evaluation of three-dimensional (3D) hydrogel thin films used to measure protein binding (antigenicity) and antibody functionality in a microarray format . Protein antigenicity was evaluated using the protein toxin, staphylococcal enterotoxin B (SEB), as a model on highly crosslinked hydrogel thin films of polyacrylamide and on two-dimensional (2D) glass surfaces . Covalent crosslinking conditions were optimized and quantified . Interrogation of the modified 3D hydrogel was measured both by direct coupling of a Cy5-labeled SEB molecule and Cy5-anti-SEB antibody binding to immobilized unlabeled SEB . Antibody functionality experiments were conducted using three chemically modified surfaces (highly crosslinked polyacrylamide hydrogels, commercially available hydrogels and 2D glass surfaces) . Cy3-labeled anti-mouse IgG (capture antibody) was microarrayed onto the hydrogel surfaces and interrogated with the corresponding Cy5-labeled mouse IgG (antigen) . Five different concentrations of Cy5-labeled mouse IgG were applied to each microarrayed surface and the fluorescence quantified by scanning laser confocal microscopy . Experimental results showed fluorescence intensities 3-10-fold higher for the 3D films compared to analogous 2D surfaces with attomole level sensitivity measured in direct capture immunoassays . However, 2D surfaces reported equal or greater sensitivity on a per-molecule basis . Reported also are the immobilization efficiencies, inter-and intra-slide variability and detection limits. J Surg Res, 2004 Nov, 122(1), 103 - 12 Depletion of activated Vbeta8+ T cells disrupts bispecific antibody directed antitumor immunity; McConnell EJ et al.; INTRODUCTION: Activation of Vbeta8+ T cells with superantigen staphylococcal enterotoxin B (SEB) and use of an antitumor, anti-CD3 bispecific antibody (BsAb) leads to tumor protective immunity . We hypothesize that Vbeta8+ T-cell activation in combination with BsAb is crucial for tumor protective immunity in this model . METHODS: Adolescent C3H/HeN mice were intravenously injected with syngeneic CL62 melanoma to establish pulmonary metastasis . Three days after establishing pulmonary metastasis, predominantly Vbeta8+ T cells are activated with 50 mug of intraperitoneal superantigen SEB . T cells were depleted at different time points in relation to SEB administration to assess the effect on protective immunity against a second tumor challenge . RESULTS: Protective immunity is significantly (P < 0.008) decreased when Vbeta8+ depletion occurs 6 h after SEB injection, as growth of rechallenged CL62 melanoma occurred in 43% . Protective immunity is present at all other time points when mice survive Vbeta8+ T-cell depletion . Survival of animals treated with SEB/BsAb (82%) is significantly better (P < 0.002) than with SEB alone (60%) or nontreated control (0%) . Survival when Vbeta8+ T-cell depletion occurred at 6 h and 48 h post-SEB is 72% and 77%, respectfully, and is statistically indistinguishable (P < 0.232 and P < 0.602) . If T-cell depletion was conducted before SEB administration, however, the combination of SEB and BsAb did not result in significant protective immunity . T-cell depletion before the use of SEB alone, without BsAb, failed to result in significant protective immunity . CONCLUSIONS: Depletion of Vbeta8+ T cells 6 h after activation disrupts the development of protective immunity. Biochem Biophys Res Commun, 2004 Dec 3, 325(1), 344 - 52 alpha-Hemolysin-induced dephosphorylation of EGF receptor of A431 cells is carried out by rPTPsigma; Vijayvargia R et al.; Earlier we have shown that the epidermal growth factor receptor was unable to retain its phospho Tyr signal after the assembly of staphylococcal alpha-hemolysin (alpha-HL) . However, the nature of the protein tyrosine phosphatase (PTPase) or its identity is not known . In this report, we demonstrate that the alpha-HL elevates the activity of receptor like protein tyrosine phosphatase sigma (rPTPsigma) . The alpha-HL induced dephosphorylation is prominent only in intact A431 cells . The PTPase activity is not inhibited if the alpha-HL treatment precedes PTPase inhibitor treatments . The anti-EGFr immunoprecipitates have exhibited higher PTPase activity after alpha-HL treatment of A431 cells . Interestingly, PTPase activity of anti-EGFr immunoprecipitates from the A431 cells expressing the antisense message of rPTPsigma has not increased despite alpha-HL treatment, confirming the role of rPTPsigma in the dephosphorylation of EGFr . The studies presented here will be useful in understanding the process of signal modulation by the assembly of alpha-HL. Eur J Cardiothorac Surg, 2004 Nov, 26(5), 897 - 900 A prospective study of autologous 'blood patch' pleurodesis for persistent air leak after pulmonary resection; Lang-Lazdunski L et al.; OBJECTIVE: To evaluate the efficacy and risks of autologous 'blood patch' pleurodesis in patients with persistent air leak after pulmonary resection . METHODS: All patients operated on by a single surgeon between January 2002 and January 2004 and presenting with a persistent air leak after pulmonary resection have been treated by the autologous blood patch pleurodesis technique . Fifty millilitres of autologous blood were injected through the chest tube that was then rinsed, clamped for 30 min and then unclamped and placed back to water seal . RESULTS: We have obtained a 100% success rate in 11 patients with persistent air leak who have been treated with this technique over a 2-year period . Most air leaks (72.7%) ceased within 12 h of blood injection . No patient developed empyema, but two patients developed fever and pleural fluid grew Staphylococcus after blood pleurodesis . At 3-month follow-up, all patients were well and their lungs were expanded fully . CONCLUSIONS: In our experience a single injection of 50 ml of blood is sufficient to seal persistent air leaks in less than 48 h . Although highly effective, the autologous blood patch pleurodesis technique should not be used in patients with incomplete lung re-expansion or positive pleural fluid culture to minimize the risk of empyema. Transplant Proc, 2004 Sep, 36(7), 2101 - 3 C4d-positive acute humoral renal allograft rejection: rescue therapy by immunoadsorption in combination with tacrolimus and mycophenolate mofetil; Liu M et al.; OBJECTIVE: We investigated the efficacy of immunoadsorption (IA) in combination with tacrolimus (FK506) and mycophenolate mofetil (MMF) rescue therapy for C4d-positive acute humoral rejection (AHR) of renal transplants . METHODS: Six of 185 cadaveric renal allograft recipients developed AHR at a mean of 4.8 +/- 0.8 days after the operation . C4d deposits were observed in peritubular capillaries (PTC) with accumulation of granulocytes . IA with staphylococcal protein A and FK506-MMF combination therapy were administered . RESULTS: After treatment with IA for 6.3 +/- 1.03 sessions combined with FK506 (0.14 to 0.16 mg.kg(-1).d(-1)) and MMF (1.5 g/d) therapy, renal function recovered in all the patients . The mean duration of treatment to a serum creatinine decrease was 14 +/- 2.9 days . The pre-IA panel reactive antibody reactivity (PRA) peaked at 50.2% +/- 6.1%, and was significantly reduced to 8.3% +/- 2.9% after IA . In four of six patients repeat allograft biopsy revealed a remission of AHR . With a mean follow-up of 18.8 +/- 5.46 months, patient and allograft survival are 100% and renal function remains stable with a mean serum creatinine of 1.2 +/- 0.22 mg/dL . CONCLUSION: The optimal treatment for alloantibody-mediated AHR remains uncertain . Our findings suggest that a therapeutic approach combining IA and FK506-MMF rescue improves the outcome of AHR. Am J Cardiol, 2004 Nov 1, 94(9), 1198 - 9 Role of troponin I in active infective endocarditis; Watkin RW et al.; Infective endocarditis (IE) remains a disease associated with high morbidity and mortality rates . In this pilot study, the role of troponin I in IE was assessed . Myocardial involvement distal to the site of infection in IE has been previously described . Elevated troponin was demonstrated in 11 of 15 patients diagnosed with the condition . Patients diagnosed with staphylococcal endocarditis were more likely to have elevated troponin (3 of 3 patients) . Patients with elevated troponin I were not more likely to need valve replacement . Troponin I levels did not predict perivalvular extension . It is hypothesized that elevated troponin I is a reflection of myocardial involvement. Nephrol Nurs J, 2004 Sep-Oct, 31(5), 534 - 7 Use of tissue plasminogen activator in peritoneal dialysis catheters: a literature review and one center's experience; Zorzanello MM et al.; There have been anecdotal reports of the use of tPA for obstructed peritoneal dialysis catheters in both adults and children . This manuscript reviews the literature and summarizes common elements of the procedures used for tPA administration in peritoneal dialysis catheters . The Gambro New Haven experience with administration of tPA (8 mgs in 10 ml of sterile water injected into the catheter and allowed to dwell for 1 hour) in 29 cases of catheter obstruction in 18 patients is presented . Patency was restored in 24 instances with no adverse effects . In the 5 cases that did not respond, the primary cause of poor drain was catheter malposition in 2, constipation in 2, and adhesions in 1 . tPA was also administered to 5 patients with relapsing peritonitis; 3 patients, all with Staphylococcus epidermidis, recovered and did not experience further recurrence. Pediatr Blood Cancer . 2004 Oct 28; {Epub ahead of print} Treatment of pediatric febrile neutropenia in the era of vancomycin-resistant microbes; Kline RM et al.; PURPOSE: The increasing frequency of Gm(+) infections in febrile neutropenic (FN) patients has resulted in increased use of vancomycin (VN) . Likely as a result, VN-resistant Enterococcus (VRE) has become a significant concern in FN patients . We sought to understand how the emergence of VN resistant microbes has changed the antibiotic management of pediatric FN . METHODS: A questionnaire was distributed by e-mail to responsible investigators of the Children's Oncology Group . RESULTS: One hundred and thirty responses were analyzed . Forty-four percent initially used monotherapy, with 82% of those using ceftazidime . Twenty-seven used VN with another agent, generally ceftazidime . After the emergence of VRE and VN-resistant staphylococcus (VRS), monotherapy increased to 58% . Ceftazidime continued to be most frequently used . There was a 57% reduction in the use of VN with 88% of centers not currently using VN in their initial treatment of FN . Forty-seven percent of the centers that continue to use VN have VRE, while 90% that have discontinued its use have VRE/VRS . CONCLUSIONS: Ours is the first study to survey current practices in the treatment of pediatric FN and to document changes in practice patterns due to emerging antibiotic resistance patterns . We demonstrate increased use of monotherapy for FN, and a 57% decrease in the use of VN . Local considerations influence antibiotic choices with a significant difference in VRE prevalence between those centers that continue to use VN as compared to those that have discontinued it . (c) 2004 Wiley-Liss, Inc. Bull Soc Belge Ophtalmol, 2004, (293), 47 - 53 Chronic granulomatous disease and peripheral ulcerative keratitis: a rare case of recurrent external ocular disease; Leroux K et al.; CLINICAL REPORT: We report a case of a 29-year-old lady, with known Chronic Granulomatous Disease, who presented with an acneiform scarring facial and trunk eruption and sore red eyes . Slitlamp examination showed limbal granulomas and adjacent peripheral ulcerative keratitis . DISCUSSION: The authors are considering the possible causes of the keratitis . As there were no signs of blepharitis at the time of initial presentation, the keratitis was most likely mediated by the adjacent limbal granuloma, and not due to Staphylococcal hypersensitivity . CONCLUSION: Although ocular involvement in CGD has been described before, this is the first article that describes limbal granulomata and peripheral ulcerative keratitis . Multidisciplinary management with longstanding oral antibiotic treatment, and topical combined antibiotic-steroid treatment were required to bring the condition under control. J Ethnopharmacol, 2004 Dec, 95(2-3), 229 - 33 Evaluation of the antioedematogenic, free radical scavenging and antimicrobial activities of aerial parts of Tillandsiastreptocarpa Baker - Bromeliaceae; Delaporte RH et al.; The crude methanolic extract of the aerial parts of Tillandsiastreptocarpa was investigated for their acute toxicity and antioedematogenic, antioxidant and antimicrobial activities . Also, the antioedematogenic activity of the hexane fraction resulting from the partition of the crude methanolic extract was evaluated . The methanolic extract and the hexane fraction showed significant (P < 0.05) inhibition of ear oedema, observed at 2 mg/ear in the croton oil-induced mice ear oedema test . In the 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging test, a high reactivity and potent antioxidant effect (IC(50) = 0.0056%, w/v) were observed for the methanolic extract . The antimicrobial activity assay showed that the crude methanolic extract was inactive toward Escherichiacoli, Staphylococcusaureus, Pseudomonasaeruginosa, Bacillussubtilis, Candidaalbicans, C . parapsilosis, C . krusei and C . tropicalis (MIC > 500 microg/ml) . The methanolic extract showed no toxic effect on mice at a single dose of 2000 mg/kg (p.o) . Common side effects including mild diarrhoea, loss of weight and depression were not recorded . The compounds cycloartenol, 4',5-dihydroxy-3',7-dimethoxyflavanone and a mixture of the steroids stigmasterol, beta-sitosterol and campesterol, were isolated from the hexane fraction and identified by spectroscopic methods. Wien Klin Wochenschr, 2004, 116 Suppl 2, 60 - 3 Abscess formation within cerebellar metastatic carcinoma--report of two cases and review of the literature; Kovacic S et al.; BACKGROUND: The occurrence of an abscess in conjunction with a tumor in the brain is very rare . Only presumptions exist about their origin and manner of dissemination . Preoperative discrimination between a brain tumor with cystic degeneration and a brain abscess within a tumor may be difficult or even impossible . The purpose of this report is to demonstrate the difficulty of such discrimination using conventional CT diagnostics alone . METHODS: Two patients with abscess formation in association with metastatic carcinoma in the cerebellum are presented and compared with similar cases in the literature . The etiology and the route of dissemination are discussed . RESULTS: In our first patient with previously diagnosed lung carcinoma and a CT showing suspected cerebellar metastasis only, an abscess caused by Propionibacterium acnes was found in the cerebellum and treated surgically . No tumor was recognised during the operation . Post-mortem examination six weeks later revealed the coexistence of remnants of a chronic abscess as well as metastatic lung carcinoma at the operation site . The second patient presented with an enhanced, ring-like cystic cerebellar lesion . During surgery, a purulent exudate with a coagulase-negative type of Staphylococcus species was found within the metastatic carcinoma of unknown origin . Both were radically excised and the patient recovered well . Neither patient had a history of previous infection and the pathway for abscess formation in both patients remained unclear . About 30 cases of abscesses associated with intracranial neoplasms were found in the literature . Including our own report, only three cases of abscesses within metastatic carcinoma have been published . CONCLUSIONS: Modern diagnostic tools reported to differentiate more reliably between an abscess and a tumor are diffusion-weighted MR and proton MR spectroscopy techniques . Brain imaging using CT alone may not reliably demonstrate both coexisting lesions or differentiate between them . It is important to know that a metastatic brain lesion can occur in association with a brain abscess and that tissue sampling for pathological as well as microbiological testing is of crucial importance for optimal therapy for both lesions. Int Immunol, 2004 Dec, 16(12), 1751 - 60 Epub 2004 Oct 25. Staphylococcal enterotoxins condition cells of the innate immune system for Toll-like receptor 4 stimulation; Rossi RJ et al.; In this report we examined overlap between superantigen (SAg) and Toll-like receptor 4 (TLR4) stimulation of the innate immune system . Before in vivo stimulation we found that mouse splenic DCs expressed unexpectedly low levels of surface TLR4 compared to macrophages . In response to LPS, TLR4 gene expression in fractionated spleen cells was downregulated . By comparison, surface TLR4 staining with the Sa15-21 mAb showed little downregulation, and the anti-TLR4 MTS510 mAb showed decreased staining, suggesting that LPS was bound to TLR4 at the time points examined . Interestingly, SAg stimulation induced decreased TLR4 staining as measured by the MTS510 mAb, even though the TLR4 gene was not downregulated . Nevertheless, LPS potently induced DCs to produce TNF and IL-12, but SAg did not, even though they efficiently activated DCs . Notwithstanding, in vivo stimulation with staphylococcal enterotoxin SAg conditioned the innate immune system to hyper-respond to various pathogen-associated molecular patterns (PAMPs) . Specifically, pre-priming with SAg enhanced LPS-mediated DC synthesis of TNF and IL-12 . Thus, SAgs may exert their pathogenesis on the host by conditioning DCs, in a T cell activation dependent manner to potentiate responses to PAMPs. Ann Plast Surg, 2004 Nov, 53(5), 420 - 4 Retrospective case review of capsular contracture after two-stage breast reconstruction: is colonization of the tissue expander pocket associated with subsequent implant capsular contracture? Macadam SA, Clugston PA, Germann ET. Periprosthetic capsular contracture is a common problem associated with implant-based breast reconstruction . The purpose of this study was to determine if bacterial colonization of the tissue expander contributes to contracture of the permanent implant . Medical records were reviewed for 86 patients (124 tissue expanders) between 1997 and 2001 in 1 institution . Three specimens taken from the expander were cultured . The overall incidence of colonization was 42.7%; 49.4% (38.8-60.0) of immediate and 28.2% (14.1-42.3) of delayed expanders had at least 1 positive culture site (P = 0.043) . The most common organisms were Propionibacterium acnes (57.6%), Staphylococcus epidermidis (31.0%), and Peptostreptococcus (5.8%) . Statistical analysis revealed no significant difference between colonization of the expander and capsular contracture of the permanent prosthesis (P = 0.59) . 45.8% (25.9-65.8) of breasts irradiated preoperatively developed contracture versus 14% (7.2-20.8) with no irradiation (P = 0.0013) . These results suggest that colonization of the expander occurs frequently, irradiation predisposes to contracture, and colonization did not contribute to secondary implant contracture in this study population. Infect Immun, 2004 Nov, 72(11), 6733 - 7 In vitro and in vivo evaluation of staphylococcal superantigen peptide antagonists; Rajagopalan G et al.; Superantigen peptide antagonists failed to block T-cell activation and cytokine production as well as toxic shock induced by staphylococcal enterotoxin B (SEB) in HLA class II transgenic mice . They also failed to inhibit the binding of SEB to HLA class II molecules as well as activation of human T lymphocytes in vitro. Infect Immun, 2004 Nov, 72(11), 6237 - 44 SdrX, a serine-aspartate repeat protein expressed by Staphylococcus capitis with collagen VI binding activity; Liu Y et al.; Staphylococcus capitis (S . capitis) has been implicated in a large proportion of coagulase-negative staphylococcal infections in very-low-birth-weight infants . To identify potential therapeutic targets, the S . capitis genome was probed for the presence of genes encoding microbial surface components recognizing adhesive matrix molecules (MSCRAMM) . By using Southern blot analysis, an S . capitis gene, designated sdrX, that contained sequence motifs consistent with the Sdr family of MSCRAMM proteins was identified . By using monospecific antisera in Western blot and flow cytometry, SdrX was demonstrated to be expressed on the surface of S . capitis . Human collagen type VI was found to bind both the recombinant A domain of SdrX and viable S . capitis expressing SdrX . SdrX is the first collagen-binding Sdr protein described and is the first MSCRAMM protein identified in S . capitis. J Cardiol, 2004 Sep, 44(3), 93 - 100 {Surgical treatment of active infective endocarditis: analysis of 20 years' experience}; Imanaka-Hasegawa M et al.; OBJECTIVES: Changes in perioperative condition and outcomes of surgically treated patients with active infective endocarditis were evaluated during the last 20 years . METHODS: Between 1983 and 2002, 132 patients with active infective endocarditis underwent surgery at Saitama Medical School . Changes in frequency, pathogens, clinical features, surgical results, and perioperative treatment were compared between four periods of 5 years . RESULTS: The percentage of surgery for infective endocarditis remained almost the same among all cardiovascular procedures . Staphylococcal infective endocarditis increased significantly (p < 0.01), and prosthetic valve infective endocarditis and periannular abscess became more common . Surgery tended to be performed in severely ill patients significantly more frequently (p < 0.01) . If all patients were included, hospital mortality did not decrease significantly, at 1.7% in stable patients, but 50% in critically ill patients . Intensive care unit stay became relatively longer . Recurrent infection was observed significantly more frequently in critically ill patients and in patients with prosthetic valve infective endocarditis . Patients were referred for surgery following diagnosis and underwent surgery at increasingly more appropriate timing . However, the diagnosis of infective endocarditis took 1.5 months to establish regardless of the patient's condition or the clinical outcome . Moreover, antibiotics were administered orally in around 90% and intravenously in nearly 70% of the patients without microbiological tests, and negative cultures remained very frequent . CONCLUSIONS: Critically ill patients underwent surgery increasingly more frequently, and surgical outcomes remained unsatisfactory over the last 20 years . Early diagnosis and avoidance of premature antibiotic therapy may be important for future improvement. Emerg Infect Dis, 2004 Sep, 10(9), 1544 - 9 Laboratory exposures to staphylococcal enterotoxin B; Rusnak JM et al.; Staphylococcal enterotoxins are 23- to 29-kDa polypeptides in the bacterial superantigen protein family . Clinical symptoms from intoxication with staphylococcal enterotoxins vary by exposure route . Ingestion results in gastrointestinal symptoms, and inhalation results in fever as well as pulmonary and gastrointestinal symptoms . Review of occupational exposures at the U.S . Army Medical Research Institute of Infectious Diseases from 1989 to 2002 showed that three laboratory workers had symptoms after ocular exposure to staphylococcal enterotoxin B (SEB) . Conjunctivitis with localized cutaneous swelling occurred in three persons within 1 to 6 hours after exposure to SEB; two of these persons also had gastrointestinal symptoms, which suggests that such symptoms occurred as a result of exposure by an indirect cutaneous or ocular route . Ocular exposures from SEB resulting in conjunctivitis and localized swelling have not previously been reported . Symptoms from these patients and review of clinical symptoms of 16 laboratory-acquired inhalational SEB intoxications may help healthcare workers evaluate and identify SEB exposures in laboratory personnel at risk. J Immunol, 2004 Nov 1, 173(9), 5810 - 7 CC chemokine ligand 18, an atopic dermatitis-associated and dendritic cell-derived chemokine, is regulated by staphylococcal products and allergen exposure; Pivarcsi A et al.; Atopic dermatitis is a chronic inflammatory skin disease with a steadily increasing prevalence . Exposure to allergens or bacterial superantigens triggers T and dendritic cell (DC) recruitment and induces atopic skin inflammation . In this study, we report that among all known chemokines CCL18/DC-CK1/PARC represents the most highly expressed ligand in atopic dermatitis . Moreover, CCL18 expression is associated with an atopic dermatitis phenotype when compared with other chronic inflammatory skin diseases . DCs either dispersed within the dermis or clustering at sites showing perivascular infiltrates are abundant sources of CCL18 . In vitro, microbial products including LPS, peptidoglycan, and mannan, as well as the T cell-derived activation signal CD40L, induced CCL18 in monocytes . In contrast to monocytes, monocyte-derived, interstitial-type, and Langerhans-type DCs showed a constitutive and abundant expression of CCL18 . In comparison to Langerhans cells, interstitial-type DCs produced higher constitutive levels of CCL18 . In vivo, topical exposure to the relevant allergen or the superantigen staphylococcal enterotoxin B, resulted in a significant induction of CCL18 in atopic dermatitis patients . Furthermore, in nonatopic NiSO4-sensitized individuals, only relevant allergen but not irritant exposure resulted in the induction of CCL18 . Taken together, findings of the present study demonstrate that CCL18 is associated with an atopy/allergy skin phenotype, and is expressed at the interface between the environment and the host by cells constantly screening foreign Ags . Its regulation by allergen exposure and microbial products suggests an important role for CCL18 in the initiation and amplification of atopic skin inflammation. Biosens Bioelectron, 2004 Oct 15, 20(3), 585 - 91 A staphylococcal enterotoxin B magnetoelastic immunosensor; Ruan C et al.; A magnetoelastic immunosensor for detection of staphylococcal enterotoxin B (SEB) is described . The magnetoelastic sensor is a newly developed mass/elasticity-based transducer of high sensitivity having a material cost of approximately $0.001/sensor . Affinity-purified rabbit anti-SEB antibody was covalently immobilized on magnetoelastic sensors, of dimensions 6 mm x 2 mm x 28 microm . The affinity reaction of biotin-avidin and biocatalytic precipitation are used to amplify antigen-antibody binding events on the sensor surface . Horseradish peroxidase (HRP) and alkaline phosphatase were examined as the labeled enzymes to induce biocatalytic precipitation . The alkaline phosphatase substrate, 5-bromo-4-chloro-3-indolyl phosphate (BCIP) produces a dimer, which binds tightly to the sensor surface, inducing a change in sensor resonance frequency . The biosensor demonstrates a linear shift in resonance frequency with staphylococcal enterotoxin B concentration between 0.5 and 5 ng/ml, with a detection limit of 0.5 ng/ml. Immunology, 1989 Jan, 66(1), 8 - 13 Herpes simplex type 1-induced Fc receptor binds to the Cgamma2-Cgamma3 interface region of IgG in the area that binds staphylococcal protein A; Johansson PJ et al.; The binding site of immunoglobulin G (IgG) to herpes simplex virus (HSV) type 1-induced Fc receptor was investigated using human IgG Fc intermediate (Fc(i)) fragments, fragment D of staphylococcal protein A (SPA) and chemically modified human IgG . Human IgG Fc(i) fragment composed of one Cgamma2 and two Cgamma3 domains, bound strongly to HSV-1-infected cells . Fragment D, a monovalent subunit of SPA, inhibited the binding of radiolabelled human IgG Fc fragments to the HSV Fc receptor . Reductively methylated human IgG reacted equally well to HSV-infected cells, as did chemically unmodified IgG in contrast to N-acetylimidazole-modified and diethylpyrocarbonate-modifed human IgG, which were unreactive . These results suggest a similar b