Staphylococcus

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 binding site on human IgG for SPA and the HSV-1 Fc receptor with involvement of the amino acid residues Tyr and His but not Lys . The similarities of binding sites on the IgG molecule for the HSV-1 Fc receptor and rheumatoid factors (RF) may be important for understanding the mechanism of RF production in rheumatoid arthritis or other disease states.

Am J Ophthalmol, 2004 Oct, 138(4), 678 - 9
Effect of chronic systemic use of trimethoprim-sulfamethoxazole in the conjunctival bacterial flora of patients with HIV infection; Fontes BM et al.; PURPOSE: To investigate the effect of orally administered trimethoprim-sulfamethoxazole (TMP-SMZ) on the prevalence, species distribution, and resistance of the conjunctival bacterial flora in adults with human immunodeficiency virus (HIV) infection . DESIGN: Cross-sectional study, with clinical and experimental laboratory investigation . METHODS: Samples from the inferior conjunctival fornix were collected and submitted for culture to evaluate aerobic flora . RESULTS: Sixty samples were collected . Negative cultures were found in 17 (56.7%) eyes of the TMP-SMZ group and in 10 (33%) of the control group (P = .036) . All Staphylococcus species isolates in the TMP-SMZ group were resistant to the drug, whereas 50% of the control group presented this finding (P = .025) . In the study group, all bacteria were resistant to TMP-SMZ, compared with only 47% of the microorganisms in the control group . CONCLUSION: Orally administered TMP-SMZ in patients with HIV infection seems to exert a selection pressure in the microorganisms present on the conjunctiva.

Mol Cell Probes, 2004 Dec, 18(6), 379 - 82
A rapid and sensitive magnetic bead-based immunoassay for the detection of staphylococcal enterotoxin B for high-through put screening; Alefantis T et al.; Staphylococcal enterotoxin B (SEB) is one of many toxins produced by the Gram-positive bacterium Staphylococcal aureus . While SEB is known as the causative agent of certain food poisonings it is also considered a biological Select Agent . Thus, rapid and accurate identification of SEB during either surveillance or in response to a biothreat is critical to the mitigation of the suspect agent . This report presents an improved method for the detection of SEB based on a SEB-specific, two-antibody system where one antibody was bound to a magnetic bead particle while the other was labeled with Alexa fluor 647 . The assay consisted of one incubation period for 30 minutes where all reagents necessary to detect SEB were included . Using this assay 100 pg of recombinant purified SEB, as well as SEB from the culture supernatant of several strains of methicillin-resistant S . aureus were detected with fidelity . This assay presents improvements over current assays in terms of a combination of the reduction in assay time length, assay sensitivity, ease of use, and application to automated high-throughput analysis . Additionally, this assay can be easily modified to detect a wide range of proteins and whole organisms.

J Antimicrob Chemother, 2004 Dec, 54(6), 1103 - 8 Epub 2004 Oct 14.
Treatment of experimental osteomyelitis by liposomal antibiotics; Kadry AA et al.; OBJECTIVES: Traditional antibiotic therapy of staphylococcal osteomyelitis by a single drug or a drug combination is ineffective in producing complete sterilization of infected bones . The aim of this study was to develop a non-traditional delivery system of antibiotics for treatment of chronic experimental osteomyelitis . METHODS: In the current work, ciprofloxacin and vancomycin were encapsulated in a cationic, anionic or neutral liposomal formulation . For prolonged circulation in serum, liposomal dispersions (<100 nm in diameter) were sonicated for different times (20, 40, 60 or 80 s), and tested for antibacterial activities . RESULTS AND CONCLUSIONS: Liposomes sonicated for 40 s gave the highest antibacterial activities in vitro . Since cationic liposomes trapped the highest percentage of antibiotics, and enhanced antibacterial activity above that of the free drugs, they were used for therapeutic trials to treat chronic staphylococcal osteomyelitis induced in rabbits . Therapeutic trials with antibiotics given intravenously revealed that, free ciprofloxacin or vancomycin given alone for 14 days was ineffective in sterilizing bone . Combination therapy with free ciprofloxacin and vancomycin for 14 days was more effective . However, this group showed renal dysfunction and severe diarrhoea, which resulted in loss of 33.3% of treated animals . Treatment with liposomal forms of either drug for 7 days was ineffective . Meanwhile, combination therapy in liposomal form for 7 days was more effective . Complete sterilization of bone tissues on cultures (100% cure) was obtained only in the group treated for 14 days with the combination of both drugs in liposomal form . Moreover, liposomal formulations showed much lower nephrotoxicity and a lower incidence of severe diarrhoea than that induced by free drugs.

Biochem Biophys Res Commun, 2004 Nov 19, 324(3), 1124 - 9
Assembly of alpha-hemolysin on A431 cells leads to clustering of Caveolin-1; Vijayvargia R et al.; Assembly and penetration of 14-strand beta-barrel of staphylococcal alpha-hemolysin (alpha-HL) is an intriguing phenomenon due to its water soluble property . alpha-HL interacts with the Caveolin-1 of A431 cells for its rapid assembly . A nine amino acid, non-hydrophobic peptide derived from alpha-HL has been shown to block the interaction of alpha-HL with the scaffolding domain of Caveolin-1 . alpha-HL's presence was also detected in the Caveolin-1 enriched membrane fractions isolated by ultracentrifugation . Moreover, alpha-HL co-precipitates with Caveolin-1 specifically . In a time-dependent process, alpha-HL associates with the Caveolin-1 and co-localizes with Caveolin-1 that results in an extensive clustering of Caveolin-1 at cell-cell contacts . Mutants of alpha-HL devoid of Caveolin-1 binding motif failed to assemble into heptameric oligomers on the surface of A431 cells . Our data suggest that the conformational changes required to form the heptameric assembly might be triggered at the Caveolin-1 binding motif of alpha-HL.

Expert Rev Anti Infect Ther, 2004 Aug, 2(4), 575 - 87
Treatment of staphylococcal scalded skin syndrome; Patel GK; Humans are a natural reservoir for Staphylococcal aureus . Colonization begins soon after birth and predisposes to infection . S . aureus is one of the most common causes of skin infection, giving rise to folliculitis, furunculosis, carbuncles, ecthyma, impetigo, cellulitis and abscesses . In addition, S . aureus may cause a number of toxin-mediated life-threatening diseases, including staphylococcal scalded skin syndrome (SSSS) . Epidermolytic toxins released by certain S . aureus strains cause SSSS by cleaving the epidermal cell adhesion molecule, desmogelin-1, resulting in superficial skin erosion . Recent experiments have revealed similarities in the pathophysiology of SSSS and pemphigus foliaceus, an autoimmune disorder that is characterized by antibodies targeting the same epidermal attachment protein . SSSS typically affects neonates and infants but may also occur in predisposed adults . It is painful and distressing for the patient and parents, although most cases respond to antibiotic treatment . Mortality is low in infants but can be as high as 67% in adults, and is dependent on the extent of skin involvement and the comorbid state . Thus, the management of adults who develop SSSS remains a major therapeutic challenge . The antibody response against the toxins neutralizes their effect and prevents recurrence or limits the effects to the area of infection, which is known as bullous impetigo.

Am Surg, 2004 Sep, 70(9), 787 - 92
Infectious complications after hepatic resection; Garwood RA et al.; The purpose of this study was to assess the characteristics of surgical infections after hepatic resection (HR) to identify factors accounting for increased postoperative mortality . Advances in operative technique and care have decreased morbidity and mortality after HR . However, infections after HR continue to be a major contributor to postoperative morbidity and mortality . All HR done during a 7-year period were analyzed and compared to our prospective surgical infection database . Factors contributing to infectious complications and mortality were identified . HR (n = 207) were performed with an overall mortality of 5.8 per cent . Nine patients (3.3%) had 18 infections; 6 (60%) had multiple infection sites, most commonly the peritoneum, blood, or wound . Three infected patients died . Lung and line infections occurred in 2 (67%) infection-related deaths . No single comorbidity increased postoperative infection risk, but an average of 6.7 comorbid conditions were present . All infection-related deaths were associated with ventilator-dependence . All infection-related deaths occurred after resection of a mean of four segments . Additional procedures at the time of HR, operative drains, or transfusion requirements did not impact infectious complications or mortality . Methicillin-resistant Staphylococcus sp . was isolated in all infection-related deaths . The mean time from HR to initiation of treatment was 8 days for infection survivors and 13.3 days for infection-related deaths . Infectious mortality after HR remains significant . Contributing risk factors are advanced age, multiple comorbid conditions, and extent of HR . Ventilator-dependence and delays in antibiotic therapy were associated with infectious mortality . Although gram-negative enteric infections were more common, abdominal, lung, and line infections with gram-positive cocci had higher associated mortality; especially when antibiotic resistant strains were present.

J Appl Microbiol, 2004, 97(5), 1087 - 94
Development of a multiplex PCR for the identification of Staphylococcus genus and four staphylococcal species isolated from food; Morot-Bizot SC et al.; AIMS: To develop a multiplex PCR that allows the identification of bacteria belonging to the Staphylococcus genus and in particular to the species Staphylococcus xylosus, S . saprophyticus, S . epidermidis and S . aureus isolated from food manufacturing plants . METHODS AND RESULTS: Five primer pairs were used in the multiplex PCR, one specific to the Staphylococcus genus and four specific to S . xylosus, S . saprophyticus, S . epidermidis and S . aureus species . All the 31 Staphylococcus reference strains yielded a specific PCR product with the genus-specific primers . Staphylococcus xylosus, S . saprophyticus, S . epidermidis and S . aureus gave a specific PCR fragment with the corresponding species-specific primers . No amplification with the Kocuria, Macrococcus and Micrococcus strains was observed in our conditions . This multiplex PCR was performed on 30 strains of Gram-positive cocci isolated from different workshops and fermented sausages . Among them, 28 belonged to the Staphylococcus genus and 14 were identified to S . saprophyticus, four to S . xylosus, two to S . aureus and one to S . epidermidis . CONCLUSIONS: This multiplex PCR provided reliable and repeatable PCR results . It allowed the identification of a major part of the isolates, highlighting the predominance of the S . saprophyticus species in the workshops studied . SIGNIFICANCE AND IMPACT OF THE STUDY: This tool is a useful way to screen the strains isolated from foodstuff and food environment and to monitor these species during the food processing.

Z Orthop Ihre Grenzgeb, 2004 Sep-Oct, 142(5), 611 - 7
{One-stage revision of shoulder arthroplasty in the case of periprosthetic infection}; Ince A et al.; AIM: Infection of shoulder arthroplasties is rare, but represents a potentially devastating complication . The aim of this work is to show the value of various diagnostic procedures, specify causative pathogens and present the results of one-stage revised patients . METHOD: We performed a retrospective analysis of our 16 consecutive patients with an infected shoulder arthroplasty . RESULTS: In 13 of 16 cases a causative pathogen could be established preoperatively . Staphylococcus and Propioni spp . dominated . Only 9 patients could be followed up because two died, two were lost and three patients were revised because of non-infectious complications . The follow-up time was 5.8 years (13 months-13.25 years) . The Constant-Murley score was 33.6 of 100 . Eradication of infection was achieved in all patients . In the follow-up time no reinfection has occurred . CONCLUSION: Culturing of the preoperative joint fluid aspirate and the determination of CRP provide an early diagnosing of shoulder arthroplasty infection . One-stage revision arthroplasty with radical debridement and application of antibiotics to the bone cement provide an accurate therapy of periprosthetic shoulder infection.

Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 2004 Oct, 98(4), 418 - 22
Relationship between oral bacteria and hemodialysis access infection; Shariff G et al.; OBJECTIVE: Controversy exists concerning the association between oral bacteria and infection of hemodialysis (HD) shunts . The purpose of this study was to determine the extent to which oral bacteria are associated with vascular access site infections in a group of HD patients . STUDY DESIGN: Microbial blood culture data for 87 HD patients were collected: 53 via chart review (retrospective), and 34 with newly suspected vascular access infections (prospective) . The primary outcome was the nature (i.e., species) of the bacteria recovered from blood cultures of subjects with vascular access infection, specifically those bacteria considered to be oral flora . RESULTS: The predominant bacteria reported to cause HD vascular access infections were Staphylococcus (48.6%) and Enterococcus (17.6%) species . Most infections occurred in hemodialysis catheters (89%) compared with AV grafts (11%) and AV fistulas (0%) . The subclavian venous access (41%) and the internal jugular venous access (38.9%) were more commonly infected than other sites . None of the bacteria identified by blood cultures were considered to be oral flora . CONCLUSION: The results suggest that oral bacteria rarely, if ever, cause vascular access infections in hemodialysis patients.

Inflamm Bowel Dis, 2004 Sep, 10(5), 655 - 6
Fatal staphylococcal sepsis in Crohn's disease after infliximab; Herrlinger KR et al.; We report a case of a 40-year-old woman who had received infliximab for perianal Crohn's disease . After six infusions of infliximab, the patient developed staphylococcal pneumonia resulting in fatal adult respiratory distress syndrome . The case is discussed in the context of the toxicity profile of infliximab .

Indian J Pathol Microbiol, 2004 Jan, 47(1), 85 - 9
Detection of slime production by coagulase-negative Staphylococcus: an assessment of two methods; Karthik S et al.; The detection of slime can be helpful in determining the diagnostic value of coagulase-negative Staphylococcus (CNS) . Here, we have made a comparative study of two phenotypic methods of slime production test from 28 clinically significant blood culture isolates of CNS . A total 16 out of 28 isolates (57.1%) were positive for slime by either of the tests, whereas only 5 out of 28 isolates (17.9%) were positive for slime by both the tests . The tube method detected slime in significantly more number of isolates than the spectrophotometric method (14 vs . 7; p<0.0483279) . A weak correlation was found between the tube method and the spectrophotometric method (rs=0.2474) . The tube test was found to be superior to the spectrophotometric test in terms of sensitivity, ease of performance and interpretation, and cost effectiveness.

Mol Cell, 2004 Oct 8, 16(1), 147 - 57
ChIC and ChEC; genomic mapping of chromatin proteins; Schmid M et al.; To map the genomic interaction sites of chromatin proteins, two related methods were developed and experimentally explored in Saccharomyces cerevisiae . The ChIC method (chromatin immunocleavage) consists of tethering a fusion protein (pA-MN) consisting of micrococcal nuclease (MN) and staphylococcal protein A to specifically bound antibodies . The nuclease is kept inactive during the tethering process (no Ca2+) . The ChEC method (chromatin endogenous cleavage) consists of expressing fusion proteins in vivo, where MN is C-terminally fused to the proteins of interest . The specifically tethered nucleases are activated with Ca2+ ions to locally introduce double-stranded DNA breaks . We demonstrate that ChIC and ChEC map proteins with a 100-200 bp resolution and excellent specificity . One version of the method is applicable to formaldehyde-fixed nuclei, another to native cells with comparable results . Among various model experiments, these methods were used to address the conformation of yeast telomeres.

Anal Biochem, 2004 Nov 1, 334(1), 72 - 80
Fluorescence resonance energy transfer-based detection of analytes using antiidiotypic affinity protein pairs; Renberg B et al.; A new method for specific detection of proteins based on fluorescence resonance energy transfer (FRET) using affinity proteins (affibodies) derived from combinatorial engineering of Staphylococcal protein A has been developed . Antiidiotypic affibody pairs were used in a homogeneous competitive binding assay, where the idiotypic, target-specific affibody was labeled with fluorescein and the antiidiotypic affibody was labeled with tetramethylrhodamine . Intermolecular FRET between the two fluorescent probes was observed in the antiidiotypic affibody complex, but upon addition of target protein the antiidiotypic affibody was displaced, which was monitored by a shift in the relative emission of the donor and acceptor fluorophores . The feasibility of the system was demonstrated by the detection of IgA and Taq DNA polymerase with high specificity, using two different antiidiotypic affibody pairs . Detection of Taq DNA polymerase in 25% human plasma was successfully carried out, demonstrating that the method can be used for analysis of proteins in samples of complex composition.

Arch Biochem Biophys, 2004 Nov 1, 431(1), 119 - 23
Perchlorate-induced conformational transition of Staphylococcal nuclease: evidence for an equilibrium unfolding intermediate; Maity H et al.; The sodium perchlorate-induced conformational transition of Staphylococcal nuclease has been monitored by both circular dichroism (CD) and fluorescence spectroscopy . The perchlorate-induced transition is cooperative as observed by both spectroscopic signals . However, the protein loses only about one-third of its native far-UV CD signal at high perchlorate concentrations, indicating that a significant amount of secondary structure remains in the post-transition state . The remaining CD signal can be further diminished in a cooperative manner by the addition of the strong denaturant, urea . Near-UV CD spectra clearly show that the protein loses its tertiary structure in the perchlorate-induced denatured state . The perchlorate-induced transition curves were fit to the standard two-state model and the standard free energy change and m value of the transition are 2.3kcal/mol and 1.8kcal/(molM), respectively . By comparison, the urea-induced unfolding of Staphylococcal nuclease (in the absence of perchlorate) yields an unfolding free energy change, DeltaG(0,un), of 5.6kcal/mol and an m value of 2.3kcal/(molM) . Thus, the thermodynamic state obtained in the post-transition region of perchlorate-induced conformation transition has a significantly lower free energy change, a high content of secondary structure, and diminished tertiary structure . These results suggest that the perchlorate-induced denatured state is a partially folded equilibrium state . Whether this intermediate is relevant to the folding/unfolding path under standard conditions is unknown at this time.

J Dermatol, 1977 Aug, 4(4), 123 - 8
Immunological aspects of staphylococcal skin infection; Tsuda S; Patients with staphylococcal infection of the skin were examined for immunological status . All cases examined in the present study had gamma-globulin levels within the normal range . The proliferative response of peripheral blood lymphocytes to staphylococcal antigen was significantly depressed in patients with furunculosis as compared with that in patients with common furuncle or in healthy controls . All patients with furunculosis showed higher titers of anti-Staphylococcus aureus antibody than did those with furuncle . The dissociation between cellular and humoral immunity in furunculosis seems to be explained in terms of immune deviation.

J Dermatol, 1977 Apr, 4(2), 65 - 8
Staphylococcal scalded skin syndrome . II . Serum level of anti exfoliatin and anti alpha-toxin in patients with staphylococcal scalded skin syndrome or bullous impetigo; Nishioka K et al.; In order to study the differences between staphylococcal scalded skin syndrome (SSSS) and bullous impetigo, the anti exfoliatin level was assessed in the sera from both groups of patients, and no significant difference in the level was found . However a significant difference was noted in the anti alpha-toxin levels in sera from both group of patients; that of SSSS patients was much lower than that of impetigo patients and of children in a control group . Five out of 6 patients with SSSS showed an unchaged level of anti alpha-toxin at the second examination, while an increased anti exfoliatin level was noted in 4 out of 6 SSSS patients.

J Mol Recognit, 2005 Jan-Feb, 18(1), 73 - 83
Modeling of receptor mimics that inhibit superantigen pathogenesis; Mollhoff M et al.; Staphylococcal enterotoxins SEB and SEC3 and toxic shock syndrome toxin TSST-1 act as superantigens by overstimulating the human immune system and thereby compromise host defense . The mechanism of pathogenesis is explained on the basis of superantigen binding to the MHC class II receptor on the antigen presenting cell and to the T cell receptor (TcR) on the T cell . SEB, SEC3 and TSST-1 bind as intact proteins and make contacts with the alpha1 subdomain (DRalpha) of MHC class II and Vbeta subdomain of TcR . SEB, SEC3 and TSST-1 show specificities for different TcRVbeta isoforms . We have designed three different chimeras linking the same DRalpha with different TcRVbeta isoforms to specifically target SEB, SEC3 and TSST-1 and inhibit their pathogenesis . Here, we show by molecular modeling that the DRalpha, TcRVbeta and linker of a given chimera interact with the target superantigen in a type-specific manner . An initial model of the complex is constructed on the basis of observed inter-molecular contacts between DRalpha/TcRVbeta and the superantigens . A constant temperature (300 K) 200 ps molecular dynamics is performed to sample different conformations of a chimera-superantigen complex by utilizing the flexibility of the (GSTAPPA)(2) linker while maintaining the native folds of superantigen, DRalpha and TcRVbeta and the observed intermolecular contacts . After equilibration, 100 molecular dynamic snapshots are minimized and analyzed . This provides descriptions of various pairwise interactions at the contact interface in the complex and important clues on single site mutations on the chimera that may enhance the stability of a given superantigen-chimera complex .

Biophys J, 2004 Oct, 87(4), 2299 - 309
Ion permeation through the alpha-hemolysin channel: theoretical studies based on Brownian dynamics and Poisson-Nernst-Plank electrodiffusion theory; Noskov SY et al.; Identification of the molecular interaction governing ion conduction through biological pores is one of the most important goals of modern electrophysiology . Grand canonical Monte Carlo Brownian dynamics (GCMC/BD) and three-dimensional Poisson-Nernst-Plank (3d-PNP) electrodiffusion algorithms offer powerful and general approaches to study of ion permeation through wide molecular pores . A detailed analysis of ion flows through the staphylococcal alpha-hemolysin channel based on series of simulations at different concentrations and transmembrane potentials is presented . The position-dependent diffusion coefficient is approximated on the basis of a hydrodynamic model . The channel conductance calculated by GCMC/BD is approximately 10% higher than (electrophysiologically measured) experimental values, whereas results from 3d-PNP are always 30-50% larger . Both methods are able to capture all important electrostatic interactions in equilibrium conditions . The asymmetric conductance upon the polarity of the transmembrane potential observed experimentally is reproduced by GCMC/BD and 3d-PNP . The separation of geometrical and energetic influence of the channel on ion conduction reveals that such asymmetries arise from the permanent charge distribution inside the pore . The major determinant of the asymmetry is unbalanced charge in the triad of polar residues D127, D128, and K131 . The GCMC/BD or 3d-PNP calculations reproduce also experimental reversal potentials and permeability rations in asymmetric ionic solutions . The weak anionic selectivity of the channel results from the presence of the salt bridge between E111 and K147 in the constriction zone . The calculations also reproduce the experimentally derived dependence of the reversible potential to the direction of the salt gradient . The origin of such effect arises from the asymmetrical distribution of energetic barriers along the channel axis, which modulates the preferential ion passage in different directions .

Int Immunopharmacol, 2004 Dec 15, 4(13), 1587 - 601
Differential expression of dendritic cell markers by all-trans retinoic acid on human acute promyelocytic leukemic cell line; Park HY et al.; Dendritic cells (DCs) are the most potent antigen-presenting cells (APCs) for naive T cells and play an important role in cancer immunology . All-trans retinoic acid (ATRA) is known to be a differentiating agent in the treatment of acute promyelocytic leukemia (APL) . In this study, we investigated whether ATRA can differentiate the retinoic acid (RA)-sensitive promyelocytic leukemic cell line, NB4, to DC-like cells and whether these differentiated cells can activate T cells . NB4 cells were differentiated to myeloid cells by 4, 6, and 8 days of ATRA treatment . NB4 cells up-regulated markers found in DCs, including HLA-DR, costimulatory molecules (CD80 and CD86), adhesion molecules (CD40), and chemokine receptors (CCR6) when cultured for 8 days in the presence of 1 microM ATRA . Upregulation of CD83 was also detected on the surface of ATRA-treated NB4 cells versus untreated cells . The addition of cytokines alone, such as GM-CSF or CD40 ligand, did not affect the expression of CD83 in untreated NB4 cells but they up-regulated CD83 in ATRA-treated cells . CD11b was coexpressed with CD80, CD83, and CD86 in ATRA-treated NB4 cells . In a functional assay, ATRA-treated NB4 cells stimulated T cell proliferation when challenged with Staphylococcus enterotoxin B . These results suggest that the differentiation of NB4 cells by ATRA causes the cells to express DC markers, and that ATRA-differentiated NB4 cells are able to present antigens to T cells.

Neurochem Res, 2004 Sep, 29(9), 1667 - 74
Fos expression in the rat brain after intraperitoneal injection of Staphylococcus enterotoxin B and the effect of vagotomy; Wang X et al.; The current study was designed to locate the neuronal activation in rat brain following intraperitoneal injection of Staphylococcus enterotoxin B (SEB) and observe the consequence of preliminary subdiaphragmatic vagotomy on SEB-induced brain Fos expression to clarify the role of the vagus nerve in sensation and transmission of abdominal SEB stimulation . The results showed that intraperitoneal SEB (1 mg/kg) induced a robust Fos expression in widespread brain areas . A significant increase of Fos immunoreactive cells were observed in the solitary tract nucleus, locus ceruleus, lateral parabrachial nucleus, ventrolateral part of central gray, medial amygdaloid nucleus, central amygdaloid nucleus, ventromedial part of thalamus, dorsomedial part of thalamus, hypothalamic paraventricular nucleus, lateral habenula, and lateral septum nucleus following SEB challenge . In hypothalamic paraventricular nucleus, in addition to the dense Fos expression in the parvocellular portion, some Fos-positive cells were also observed in the anterior magnocellular nucleus of the complex . Double immunofluorescence studies showed that these Fos-immunoreactive cells were mostly oxytocinergic . The results also showed that subdiaphragmatic vagotomy largely attenuated, but not totally abrogated, the brain Fos expression induced by abdominal administration of SEB . Our data suggest that peripheral SEB stimulation can induce activation of neurons in widespread brain areas and that the vagus plays a crucial role in transmitting the signal of abdominal immune stimulation to the brain.

Anal Biochem, 2004 Oct 15, 333(2), 225 - 9
Determination of protein charge by capillary zone electrophoresis; Winzor DJ et al.; The feasibility of employing classical electrophoresis theory to determine the net charge (valence) of proteins by capillary zone electrophoresis is illustrated in this paper . An outline of a procedure to facilitate the interpretation of mobility measurements is demonstrated by its application to a published mobility measurement for Staphylococcal nuclease at pH 8.9 that had been obtained by capillary zone electrophoresis . The significantly higher valence of +7.5 (cf . 5.6 from the same series of measurements) that has been reported on the basis of a "charge ladder" approach for charge determination signifies the likelihood that the latter generic approach may be prone to error arising from nonconformity of the experimental system with an inherent assumption that chemical modification or mutation of amino acid residues has no effect on the overall three-dimensional size and shape of the protein.

Indian J Pediatr, 2004 Sep, 71(9), 819 - 24
Management of septic arthritis; Shetty AK et al.; Septic arthritis in children remains a serious disease with the potential for significant systemic and musculoskeletal morbidity . Staphlococcus aureus is the most common cause of bone and joint infections in all age groups . Microbial invasion of the synovial space occurs typically results from hematogenous seeding . Diagnosis in neonates and young infants can be difficult since the clinical signs are much less specific in these age groups . Early diagnosis by needle aspiration of the affected joint and prompt initiation of appropriate antimicrobial therapy in conjunction with drainage of the affected joint is critical to avoid destruction of the articular cartilage and prevent disability . Septic arthritis in infants and children should always be managed by a pediatrician in close consultation with an orthopedic surgeon . Empiric antibiotic regimens should always include adequate anti-staphylococcal coverage . Antibiotic treatment should be started with appropriate doses of intravenous antibiotics . Switch to oral antibiotic therapy can be made when patient demonstrates clinical improvement . A minimum of 3-4 weeks of therapy is recommended . Close follow-up is warranted to monitor the growth of the affected limb until skeletal maturity.

Clin Cancer Res, 2004 Sep 15, 10(18 Pt 2), 6360S - 6S
Effect of renal cell carcinomas on the development of type 1 T-cell responses; Rayman P et al.; PURPOSE: We reported that in renal cell carcinoma patients with active disease, T-cell reactions to the tumor-associated antigens MAGE-6 and EphA2 are highly skewed toward TH2-type cytokine responses {interleukin (IL) 5} . Herein, we determined whether tumor-derived products, including gangliosides isolated from renal cell carcinoma patients, participate in the down-regulation of type 1 T-cell responses . EXPERIMENTAL DESIGN: T cells from healthy volunteers or renal cell carcinoma patients were cultured in the presence and absence of supernatants derived from renal cell carcinoma explants or with gangliosides isolated from those tumor supernatants . T cells were stimulated or not with either autologous dendritic cells pulsed with superantigen (Staphylococcus enterotoxin B) or with phorbol 12-myristate 13-acetate and ionomycin and then were assessed for type 1 or type 2 responses (cytokine production and gene expression) and apoptosis . RESULTS: Tumor supernatants efficiently inhibited the TH1-type responses {interferon (IFN) gamma} of T cells stimulated with either S . enterotoxin B or phorbol 12-myristate 13-acetate and ionomycin but had no inhibitory effect on activated T-cell production of type 2 cytokines (IL-4, IL-5, and IL-10) . Likewise, IFN-gamma mRNA and protein production were inhibited when T cells were cocultured with either renal cell carcinoma supernatant-derived gangliosides or a commercial source of purified GD1a . It was also determined that gangliosides impair type 1 responses by inducing apoptosis of activated T cells . CONCLUSIONS: We propose that renal cell carcinoma-derived tumor products such as gangliosides can induce a type 2 bias in antitumor immunity by initiating apoptosis in the IFN-gamma-producing type 1 effector cells . This represents a relevant mechanism by which renal cell carcinoma can inhibit protective antitumor immunity.

Antimicrob Agents Chemother, 2004 Oct, 48(10), 3905 - 11
Human immunodeficiency virus type 1 protease inhibitors block toll-like receptor 2 (TLR2)- and TLR4-Induced NF-kappaB activation; Equils O et al.; Coinfections with opportunistic and pathogenic bacteria induce human immunodeficiency virus (HIV) replication through microbial antigen activation of NF-kappaB . Here, we assessed whether HIV type 1 protease inhibitors (PI) block microbial antigen activation of NF-kappaB . Human microvessel endothelial cells were transiently transfected with either endothelial cell-leukocyte adhesion molecule NF-kappaB luciferase or interleukin 6 (IL-6) promoter luciferase constructs by using FuGENE 6, and they were treated with PI (nelfinavir, ritonavir, or saquinavir) prior to stimulation with the Toll-like receptor 4 (TLR4) and TLR2 ligands, with lipopolysaccharide (LPS), soluble Mycobacterium tuberculosis factor, or Staphylococcus epidermidis phenol-soluble modulin, respectively, or with tumor necrosis factor alpha (TNF-alpha) . Luciferase activity was measured by using a Promega luciferase kit . TNF-alpha release from the supernatant was measured by enzyme-linked immunosorbent assay . Cell death was assessed by lactate dehydrogenase assay . We observed that PI pretreatment blocked the TLR2- and TLR4- as well as the TNF-alpha-mediated NF-kappaB activation, in a dose-dependent manner . PI pretreatment also blocked the LPS-induced IL-6 promoter transactivation and TNF-alpha secretion . These data suggest that PI block HIV replication not only by inhibiting the HIV protease but also by blocking the TLR- and TNF-alpha-mediated NF-kappaB activation and proinflammatory cytokine production . These findings may help explain the immunomodulatory effects of PI, and they suggest an advantage for PI-containing drug regimens in the treatment of HIV-infected patients who are coinfected with opportunistic and pathogenic bacteria.

Adv Perit Dial, 2004, 20, 74 - 7
The recommendations from the International Society for Peritoneal Dialysis for Peritonitis Treatment: a single-center historical comparison; Silva MM et al.; The antibiotic treatment currently recommended by the International Society for Peritoneal Dialysis (ISPD) for peritonitis consists of a combination of a first- and a third-generation cephalosporin . The schedule formerly recommended combined a first-generation cephalosporin and an aminoglycoside . No comparison between the treatment schedules has been performed until now . We compared the effectiveness of these two regimens in peritoneal dialysis-related peritonitis at our center . From January 1999 to April 2000, we followed 107 patients in our PD clinic (period 1: 47% men; 32% with diabetes; mean age: 52 +/- 13 years) . We followed a similar number of patients from January 2002 to July 2003 (period 2: 109 patients; 54% men; 51% with diabetes; mean age: 56 +/- 18 years) . In each period, diagnosis and treatment of peritonitis were based on the recommendations of the ISPD as earlier described . Negative culture rates were similar in period 1 and period 2 (32% vs . 30%) . In both study groups, the bacteria that most commonly caused peritonitis were Staphylococcus epidermidis (period 1: 41%; period 2: 39%) and S . aureus (period 1: 27%; period 2: 18%) . Gram-positive infections occurred in 59% of patients during period 1 and in 57% during period 2 . Gram-negative infections occurred in 16% of patients during period 1 and in 18% during period 2 . We observed no significant difference in the peritonitis cure rate from period 1 to period 2 (78% vs . 83%; chi-square: 0.98; p = 0.3), but changes in the primary antibiotic schedule were necessary in 4 patients in period 1 as compared with 1 patient in period 2 . The rates of catheter removal were not significantly different during the two periods (period 1: 14%; period 2: 5%; chi-square: 2.5; p = 0.11) . Mortality was also not significantly different during the two periods (period 1: 7%; period 2: 5%; chi-square: 0.23; p = 0.62) . The two antibiotic schedules were equally effective in the treatment of peritonitis . Cost-effectiveness, impact on residual renal function, and potential development of bacterial resistance must be considered when selecting the antibiotic schedule for peritonitis treatment.

Orv Hetil, 2004 Jun 27, 145(26), 1367 - 72
{Necrotising fasciitis in hyper-IgE syndrome}; Erdos M et al.; Hyper-IgE syndrome is a rare primary immunodeficiency disease characterized by recurrent staphylococcal skin abscesses, chronic eczematoid dermatitis, pneumonia, pneumatoceles, and extreme elevation of serum IgE . The most common pathogens are S . aureus, and C . albicans . Abnormalities of dentition, bone manifestations, and connective tissue disorders are also common features of the disease . The authors report here a 19-year-old female with hyper-IgE syndrome who developed necrotising fasciitis and toxic shock syndrome . Methicillin resistant S . aureus and S . pyogenes cultured from the skin lesions . Association of hyper-IgE syndrome with necrotising fasciitis is a rarity in the medical literature . In addition to the case report, the authors describe here the major immunologic and clinical manifestations of hyper-IgE syndrome.

Am J Kidney Dis, 2004 Oct, 44(4), e50 - 8
The role of circulating immune complexes and biocompatibility of staphylococcal protein A immunoadsorption in mitomycin C-induced hemolytic uremic syndrome; Mistry B et al.; Mitomycin-induced hemolytic uremic syndrome (HUS) is a life-threatening complication of this therapy, and increased levels of circulating immune complexes and hypocomplementemia have been found in some patients . We further characterize the role of immune complexes in mitomycin-HUS by showing that removal of these complexes by immunoadsorption with staphylococcal protein A columns correlates with temporal improvement in the microangiopathic hemolytic anemia in this disorder . Two immune complexes bound to the protein A column were identified: an 11S platelet-aggregating protein and a 15S non-platelet-aggregating protein . In addition, the patient had anaphylactoid reactions at the onset of immunoadsorption similar to first-use dialysis reactions that correlated with increases in complement 3a and 5a, interleukin-1 (IL-1), IL-6, and tumor necrosis factor levels . This case suggests that platelet-aggregating and complement-fixing circulating immune complexes are, in part, a proximate cause for mitomycin C-induced HUS . Therapy for mitomycin (HUS) with protein A columns should continue until circulating immune complexes reach undetectable levels and serum complement levels return to the normal range.

Biopolymers, 2004 Oct 15, 75(3), 229 - 41
Searching for folding initiation sites of staphylococcal nuclease: a study of N-terminal short fragments; Dai J et al.; The N-terminal short fragments of staphylococcal nuclease (SNase), SNase20, SNase28, and SNase36, corresponding to the sequence regions, Ala1-Gly20, Ala1-Lys28, and Ala1-Leu36, respectively, as well as an 8-residue peptide (Ala17-Ile18-Asp19-Gly20-Asp21-Thr22-Val23-Lys24) have been synthesized . The conformational states of these fragments were investigated using CD and NMR spectroscopy in aqueous solution and in trifluoroethanol (TFE)-H(2)O mixture . SNase20 containing a sequence corresponding to a bent peptide in native SNase shows a transient population of bend-like conformation around Ala12-Thr13-Leu14 in TFE-H(2)O mixture . The sequence region of Ala17-Thr22 of SNase28 displays a localized propensity for turn-like conformation in both aqueous solution and TFE-H(2)O mixture . The conformational ensemble of SNase36 in aqueous solution includes populated turn-like conformations localized in sequence regions Ala17-Thr22 and Tyr27-Gln30 . The analysis suggests that these sequence regions, which form the regular secondary structures in native protein, may serve as the folding nucleation sites of SNase fragments of different chain lengths starting from the N-terminal end . Thus, the formation of bend- and turn-like conformations of these sequence regions may be involved in the early folding events of the SNase polypeptide chain in vitro.

Biophys J, 2004 Dec, 87(6), 3982 - 94 Epub 2004 Dec.
Stabilization of internal charges in a protein: water penetration or conformational change?
Denisov VP, Schlessman JL, Garcia-Moreno E B, Halle B.
The ionizable amino acid side chains of proteins are usually located at the surface . However, in some proteins an ionizable group is embedded in an apolar internal region . Such buried ionizable groups destabilize the protein and may trigger conformational changes in response to pH variations . Because of the prohibitive energetic cost of transferring a charged group from water to an apolar medium, other stabilizing factors must be invoked, such as ionization-induced water penetration or structural changes . To examine the role of water penetration, we have measured the 17O and 2H magnetic relaxation dispersions (MRD) for the V66E and V66K mutants of staphylococcal nuclease, where glutamic acid and lysine residues are buried in predominantly apolar environments . At neutral pH, where these residues are uncharged, we find no evidence of buried water molecules near the mutation site . This contrasts with a previous cryogenic crystal structure of the V66E mutant, but is consistent with the room-temperature crystal structure reported here . MRD measurements at different pH values show that ionization of Glu-66 or Lys-66 is not accompanied by penetration of long-lived water molecules . On the other hand, the MRD data are consistent with a local conformational change in response to ionization of the internal residues.

Support Care Cancer . 2004 Sep 15; {Epub ahead of print}
Cutaneous bacterial colonization, modalities of chemotherapeutic infusion, and catheter-related bloodstream infection in totally implanted venous access devices; Laurenzi L et al.; GOALS OF WORK . Prospective clinical study to evaluate patients suffering from solid tumor using a totally implanted venous access device (TIVAD) to determine: (1) if there is a relationship between cutaneous contamination at port insertion site and catheter-related bloodstream infection (CRBI); (2) development modalities of CRBI; (3) if there is a relationship between chemotherapy administration modalities by push/bolus versus continuous infusion and CRBI . PATIENTS AND METHODS . We studied 41 consecutive patients who needed a TIVAD positioned for chemotherapy administration by bolus/push or continuous infusion . In every patient, we performed blood cultures from blood samples from port catheters and cutaneous cultures from cutaneous tampons of the skin surrounding the implant area on the first (T0) and eight day (T1) postoperatively, after 1 month (T2), and after 3 months (T3) from insertion . MAIN RESULTS . The study was completed on 40 patients; in one case, the port was removed at T2 for septic complications . We obtained four positive blood cultures (two, 5%), two in the same patient, all caused by staphylococcus . Positive cutaneous tampons were 21 (13%) in 11 patients (27%); the four CRBI occurred in this group of patients with none in the remaining 30 patients (73%) for a total number of 120 tampons ( p<0.01) . In two cases, the same germ was isolated from both the skin and blood . None of the patients presented a local infection of the subcutaneous pocket . Positive cutaneous cultures decrease over time: T0-T2, 24-5%; T1-T3, 20-5% ( p<0.04) . There were no differences in CRBI incidence and positive cutaneous tampons between the two chemotherapy administration modalities . CONCLUSIONS . Cutaneous microbial flora has a primary role in CRBI development within TIVADs; there is a relationship between cutaneous colonization and CRBI; colonization reaches its maximum during the first days after catheterization in which the use of the system is at high risk; colonization occurs both via extraluminal and endoluminal routes; there is no difference in CRBI incidence between bolus and continuous infusion administration.

J Dermatolog Treat, 2004 Sep, 15(5), 303 - 7
Mycophenolate mofetil in pyoderma gangrenosum; Lee MR et al.; Recently, the use of mycophenolate mofetil (MMF) has expanded in dermatological practice . Three patients with PG received MMF in conjunction with prednisone and a fourth patient received MMF monotherapy daily . MMF in combination with prednisone in three of our patients and alone in our fourth patient induced healing of their ulcers . The first patient tolerated MMF apart from gastrointestinal upset, while treatment in our second patient was complicated by staphylococcal and pseudomonal sepsis . The third patient complained of palpitations and headaches.

Semin Thorac Cardiovasc Surg, 2004 Spring, 16(1), 108 - 12
Mediastinal wound infections following pediatric cardiac surgery; Huddleston CB; Deep sternal wound infections in children and neonates occur at approximately the same incidence as in adults . The organisms responsible are generally Staphylococcal species . The risk factors in adults include diabetes mellitus, the use of internal mammary artery grafts for bypass conduits, and other factors which clearly relate to age and the nature of the operations performed . In children the risk factors include leaving the sternum open following repair, nasal colonization with Staphylococcal species, need for re-exploration for bleeding, and others . Although conventional therapy for this complication is similar to what is proposed for adults, recently interest has been established in treating with debridement and primary closure of the sternum.

Curr Opin Cell Biol, 2004 Oct, 16(5), 536 - 43
Desmosomes and disease: pemphigus and bullous impetigo; Payne AS et al.; Desmosomal cadherins are the pathophysiologic targets of autoimmune or toxin-mediated disruption in the human diseases pemphigus and bullous impetigo (including its generalized form, called staphylococcal scalded skin syndrome) . Experiments exploiting the production of both pathogenic and nonpathogenic antidesmoglein antibodies in pemphigus patients' sera have afforded data that make an invaluable contribution towards identifying the functional domains of the desmogleins involved in intercellular adhesion . Conformational epitopes of antidesmoglein autoantibodies in pemphigus patients' sera and the specific cleavage site of desmoglein 1 by exfoliative toxin have been identified, implicating the N-terminal extracellular domains of the desmogleins as critical regions for controlling intercellular adhesion . Furthermore, the development of active autoimmune mouse models for pemphigus allows in vivo characterization of the disease and its pathogenesis . These studies offer new insight into the potential mechanisms of acantholysis in pemphigus and staphylococcal-associated blistering disease, with implications for the role of desmogleins in desmosomal structure and function .

Clin Diagn Lab Immunol, 2004 Sep, 11(5), 957 - 62
Response to superantigen stimulation in peripheral blood mononuclear cells from children perinatally infected with human immunodeficiency virus and receiving highly active antiretroviral therapy; McCloskey TW et al.; Our understanding of the pathogenesis of perinatal human immunodeficiency virus (HIV) infection is still evolving . We sought to characterize the response to the bacterial superantigen Staphylococcus enterotoxin B (SEB) of lymphocytes from HIV-infected children receiving treatment with highly active antiretroviral therapy (HAART) . Using the flow cytometric methodology, we quantified apoptosis, proliferation, cytokine production, and activation antigen upregulation in CD4 and CD8 T lymphocytes following in vitro stimulation of peripheral blood mononuclear cells (PBMCs) with SEB . The levels of proliferation, CD4 interleukin-2 (IL-2) production, CD8 gamma interferon (IFN-gamma) production, and upregulation of CD69 expression by cells from HIV-infected children were indistinguishable from those by cells from controls . However, stimulation with SEB dramatically decreased the ratio of resting apoptotic cells to cycling apoptotic cells in the controls but not in the patients . In addition, unstimulated spontaneous apoptosis of CD4 T cells remained greater in the patients than in the controls . The percentages of IL-2-positive CD8 T cells and IFN-gamma-positive CD4 T cells following SEB stimulation were significantly lower in the patients than in the controls . Our multiparameter approach was able to demonstrate differences in lymphocyte superantigen responsiveness in HIV-infected children receiving HAART in comparison to that in uninfected controls, notably, an apoptotic versus a proliferative response to stimulation.

J Appl Microbiol, 2004, 97(4), 719 - 30
PCR detection of staphylococcal enterotoxin genes in Staphylococcus spp . strains isolated from meat and dairy products . Evidence for new variants of seG and seI in S . aureus AB-8802; Blaiotta G et al.; AIMS: Evaluation of the occurrence of most known staphylococcal enterotoxin (SE) genes, egc (enterotoxin gene cluster) and TSST1 (toxic shock syndrome toxin 1) gene in both coagulase-positive (CPS) and coagulase-negative (CNS) staphylococcal strains isolated from meat and dairy products . METHODS AND RESULTS: Specificity and reliability of the PCR detection methods used were ascertained by using nine reference strains of Staphylococcus (S . aureus) harbouring SE genes (seA to seE; seG, seH, seI, seM, seJ, seN and seO) and egc (containing the following sequence of genes: seO, seM, seI, phient1, phient2, seN and seG) . Of 109 wild Staphylococcus spp . strains analysed, only 11 S . aureus strains were SE and/or TSST1 PCR-positive . The last 11 strains also appeared to harbour the egc . Restriction endonuclease analysis of part of the egc of both reference and wild strains showed that different variants of the egc exist . Moreover, nucleotide sequences of seG and seI indicate that the egc of the strain AB-8802 is characterized by the presence of variants of these enterotoxins (seGv and seIv) . CONCLUSIONS: The occurrence of SE genes in CNS and other non-S . aureus species isolated from Napoli-type salami, raw water buffalo milk and natural whey cultures used for mozzarella cheese manufacturing is very rare . SIGNIFICANCE AND IMPACT OF THE STUDY: During this study it was shown that at least five different egc may exist in S . aureus . A thorough study of egc polymorphism should provide further insight into the phylogenetics of the egc.

Infect Control Hosp Epidemiol, 2004 Aug, 25(8), 664 - 7
In vitro assessment of chlorhexidine gluconate-impregnated polyurethane foam antimicrobial dressing using zone of inhibition assays; Bhende S et al.; OBJECTIVE: To evaluate an antimicrobial dressing consisting of hydrophilic polyurethane foam with chlorhexidine gluconate for activity against several antibiotic-resistant clinical isolates as well as American Type Culture Collection reference strains using zone of inhibition assays . METHODS: Sterile foam samples with chlorhexidine gluconate and untreated controls were transferred onto inoculated agar plates . Plates were incubated at 35 degrees C to 37 degrees C for 24 hours and examined for zones of inhibition around the foam samples . RESULTS: Polyurethane foam with chlorhexidine gluconate showed antimicrobial activity in vitro against all of the challenge organisms including antibiotic-resistant clinical isolates . CONCLUSION: The data from this in vitro study support the hypothesis that polyurethane foam with chlorhexidine gluconate has an antimicrobial effect against antibiotic-resistant Staphylococcus and Enterococcus species, as well as Candida species.

J Exp Med, 2004 Sep 6, 200(5), 613 - 22
Determination of the critical concentration of neutrophils required to block bacterial growth in tissues; Li Y et al.; We showed previously that the competition between bacterial killing by neutrophils and bacterial growth in stirred serum-containing suspensions could be modeled as the competition between a first-order reaction (bacterial growth) and a second-order reaction (bacterial killing by neutrophils) . The model provided a useful parameter, the critical neutrophil concentration (CNC), below which bacterial concentration increased and above which it decreased, independent of the initial bacterial concentration . We report here that this model applies to neutrophil killing of bacteria in three-dimensional fibrin matrices and in rabbit dermis . We measured killing of 10(3)-10(8) colony forming units/ml Staphylococcus epidermidis by 10(5)-10(8) human neutrophils/ml in fibrin gels . The CNC was approximately 4 x 10(6) neutrophils/ml gel in the presence of normal serum and approximately 1.6 x 10(7) neutrophils/ml gel in the presence of C5-deficient serum . Application of our model to published data of others on killing of approximately 5 x 10(7) to 2 x 10(8) E . coli/ml rabbit dermis yielded CNCs from approximately 4 x 10(6) to approximately 8 x 10(6) neutrophils/ml dermis . Thus, in disparate tissues and tissuelike environments, our model fits the kinetics of bacterial killing and gives similar lower limits (CNCs) to the neutrophil concentration required to control bacterial growth.

J Mater Sci Mater Med, 1997, 8(10), 631 - 5
Conditioning fluid influences on the surface properties of silicone and polyurethane peritoneal catheters: implications for infection; Gorman SP et al.; Catheter-related infection remains a considerable problem in continuous ambulatory peritoneal dialysis (CAPD) . This study examined the adherence of clinical isolates of Staphylococcus epidermidis to commercially available polyurethane and silicone peritoneal catheters in the presence and absence of a proteinaceous conditioning film . In addition, the effects of the conditioning film on the surface properties (advancing and receding contact angles, and surface rugosity) of these biomaterials were investigated . Bacterial adherence to polyurethane and silicone catheters, pre-treated with phosphate-buffered saline (PBS) or artificial spent peritoneal dialysate (ASD) for 1 h at 37 degrees C, was examined using a radiometric (2-3H-adenine) adherence assay . The advancing and receding contact angles and the surface rugosity of ASD- and PBS-treated biomaterials were examined using a dynamic contact angle analyser and an atomic force microscope, respectively . The bacterial isolates were selected to represent high and low cell surface hydrophobicity . The hydrophobic isolate exhibited both a significantly greater rate and a significantly greater extent of adherence than the hydrophilic isolate to both catheter materials, independent of pre-treatment . In general, pre-treatment of the catheter materials with ASD significantly decreased the subsequent adherence of both isolates owing to the deposition of a conditioning film on the surface of the biomaterial . ASD treatment also decreased both the advancing and receding contact angles and the surface rugosity of both catheter materials . This study highlights the influence of both bacterial cell surface hydrophobicity and biomaterial surface conditioning films on bacterial adherence to CAPD catheters . In addition, it is recommended that the effects of proteinaceous conditioning films on biomaterial surface properties should be considered when assessing materials for medical devices and products.

J Mater Sci Mater Med, 2001 Jun, 12(6), 543 - 8
Staphylococcus epidermidis RP62A adhesion to chemically modified cellulose derivatives; Fonseca AP et al.; The adhesion of coagulase negative Staphylococcus epidermidis expressing capsular polysaccharide/adhesin (PS/A) to cellulose diacetate (CDA), as well as to primary reference low-density polyethylene, was assessed in vitro . Attached bacteria were released by gentle sonication and quantified as colony forming units . Surface free energy of cells and materials and the free energy of interaction between cells, each type of material and water molecules was calculated through contact angle measurement, also enabling the determination of materials surface hydrophobicity . The influence of CDA surface modification by deacetylation and phosphorylation on bacterial adhesion was studied . Chemical modifications of CDA by deacetylation and by phosphorylation were effective in lowering bacterial adhesion .

J Mater Sci Mater Med, 1999, 10(10/11), 621 - 7
The development of in vitro biocompatibility tests for the evaluation of intraocular biomaterials; Lloyd AW et al.; Recent developments in ocular implant technology require the in vitro evaluation of ocular compatibility in early stage development programs . This requires an understanding and appreciation of the biological interactions which occur in the ocular environment and their relevance with respect to the clinical complications associated with surgical implantation of devices . This paper describes the development of a series of clinically reflective in vitro assays for assessing the potential ocular compatibility of novel intraocular lens materials . Staphylococcus epidermidis attachment, fibrinogen adsorption, mouse embryo fibroblast 3T3 adhesion and proliferation, primary rabbit lens cell adhesion, human peripheral blood macrophage adhesion and granulocyte activation tests were employed to evaluate two widely used intraocular biomaterials poly(methyl methacrylate) (PMMA) and silicone, and a novel biomimetic phosphorylcholine-based coating (PC) . The performance of these materials in the in vitro assays was compared to their ability to reduce postoperative inflammation in vivo in a rabbit model . The results demonstrated that the in vitro assays described here are predictive of in vivo ocular compatibility . These assays offer a more relevant means of assessing the ocular compatibility of biomaterials than those presently required by the authorities for regulatory approval of medical devices and implants .

Biophys J, 2004 Nov, 87(5), 3479 - 92 Epub 2004 Sep 03.
Pressure denaturation of staphylococcal nuclease studied by neutron small-angle scattering and molecular simulation; Paliwal A et al.; We studied the pressure-induced folding/unfolding transition of staphylococcal nuclease (SN) over a pressure range of approximately 1-3 kilobars at 25 degrees C by small-angle neutron scattering and molecular dynamics simulations . We find that applying pressure leads to a twofold increase in the radius of gyration derived from the small-angle neutron scattering spectra, and P(r), the pair distance distribution function, broadens and shows a transition from a unimodal to a bimodal distribution as the protein unfolds . The results indicate that the globular structure of SN is retained across the folding/unfolding transition although this structure is less compact and elongated relative to the native structure . Pressure-induced unfolding is initiated in the molecular dynamics simulations by inserting water molecules into the protein interior and applying pressure . The P(r) calculated from these simulations likewise broadens and shows a similar unimodal-to-bimodal transition with increasing pressure . The simulations also reveal that the bimodal P(r) for the pressure-unfolded state arises as the protein expands and forms two subdomains that effectively diffuse apart during initial stages of unfolding . Hydrophobic contact maps derived from the simulations show that water insertions into the protein interior and the application of pressure together destabilize hydrophobic contacts between these two subdomains . The findings support a mechanism for the pressure-induced unfolding of SN in which water penetration into the hydrophobic core plays a central role.

Bone, 2004 Sep, 35(3), 614 - 20
Effects of activated T cells on osteoclastogenesis depend on how they are activated; Wyzga N et al.; INTRODUCTION: Activated T cells are emerging as important regulators of osteoclast function in inflammatory diseases . Both pro- and anti-resorptive properties have been described . We reasoned that this reported variability of the effects of T cells on osteoclast formation depends on how T cells are activated in vitro . METHODS: We harvested T lymphocytes from 5-week-old C57BL/6 mouse spleens . Activation was performed with anti-CD3epsilon and -CD28 Ab (Abs), concanavalin A (Con A), phytohemagglutinin (PHA), or the superantigen Staphylococcal enterotoxin A (SEA) . Osteoclastogenesis was induced by receptor activator of NF-kappaB ligand (RANKL) in the mouse macrophage cell line RAW 264.7 cells, or primary macrophage CD11b+ cells from mouse spleen . Cells were cultured with T cells or with their conditioned medium . RESULTS: Co-culture of activated T lymphocytes with RAW 264.7 cells inhibited osteoclastogenesis but only when activated by Abs . This effect was CD4+ -dependent . Conditioned medium from activated T lymphocytes with Abs consistently blocked osteoclastogenesis in RAW 264.7 and CD11b+ cells . T cells activated with SEA, Con A, and PHA had inconsistent effects on osteoclastogenesis . We then tested the role of interferon (IFN)-gamma, a known inhibitor of osteoclastogenesis, in the effects of T cells on osteoclast formation . IFN-gamma neutralizing antibody blocked the inhibitory effect of T-cell conditioned medium on osteoclastogenesis . Osteoclast precursors from IFN-gamma receptor-null mice treated with 0.1% medium from activated T cells formed osteoclasts . However, higher doses of medium inhibited osteoclastogenesis, so that we cannot exclude that other factors besides IFN-gamma may be involved . CONCLUSIONS: Available methods to activate T lymphocytes result in variable effects on osteoclastogenesis . IFN-gamma is the main factor responsible for the inhibitory effects of activated T cells on osteoclast formation.

Curr Biol, 1993 Jul 1, 3(7), 414 - 23
A method for predicting protein structure from sequence; Skolnick J et al.; Background: The ability to predict the native conformation of a globular protein from its amino-acid sequence is an important unsolved problem of molecular biology . We have previously reported a method in which reduced representations of proteins are folded on a lattice by Monte Carlo simulation, using statistically-derived potentials . When applied to sequences designed to fold into four-helix bundles, this method generated predicted conformations closely resembling the real ones . Results: We now report a hierarchical approach to protein-structure prediction, in which two cycles of the above-mentioned lattice method (the second on a finer lattice) are followed by a full-atom molecular dynamics simulation . The end product of the simulations is thus a full-atom representation of the predicted structure . The application of this procedure to the 60 residue, B domain of staphylococcal protein A predicts a three-helix bundle with a backbone root mean square (rms) deviation of 2.25-3 A from the experimentally determined structure . Further application to a designed, 120 residue monomeric protein, mROP, based on the dimeric ROP protein of Escherichia coli, predicts a left turning, four-helix bundle native state . Although the ultimate assessment of the quality of this prediction awaits the experimental determination of the mROP structure, a comparison of this structure with the set of equivalent residues in the ROP dime- crystal structure indicates that they have a rms deviation of approximately 3.6-4.2 A . Conclusion:Thus, for a set of helical proteins that have simple native topologies, the native folds of the proteins can be predicted with reasonable accuracy from their sequences alone . Our approach suggest a direction for future work addressing the protein-folding problem.

Kardiol Pol, 2004 Jun, 60(6), 535 - 40
Echocardiographic predictors of embolic events in infective endocarditis; Macarie C et al.; BACKGROUND: Systemic embolisation occurs in 22% to 50% of patients with infective endocarditis (IE) . Up to 65% of embolic events (EE) involve the central nervous system which increases the mortality rate . Several echocardiographic studies have demonstrated higher embolic rates with the increase of vegetation (VEG) dimensions and mobility . AIM: To define echocardiographic parameters which can help in identifying patients with a high risk of EE and to assess the value of transesophageal echocardiography (TEE) in predicting EE in patients with IE . METHODS: 236 patients (58% male, mean age 47.8+/-6) diagnosed with IE according to Duke criteria were followed for 3 years or until cardiac surgery . Echocardiographic parameters measured on VEG included the maximum length, thickness, the narrowest diameter, neck and mobility . RESULTS: The rate of EE was 51.27% without any significant differences with respect to gender, age, fever, anaemia, VEG site or the presence of a significant regurgitation murmur . The univariate analysis showed a significant correlation between EE and IE caused by staphylococcus, IE of the right heart, and the length as well as mobility of VEG . The only independent predictors of EE were the maximum VEG length >15 mm and the increased mobility of VEG with a maximal displacement angle >60.7 degrees . In 23% of patients EE occurred after the initiation of antibiotic treatment . VEG in this group were big and very mobile (length >15 mm, maximal angle of displacement >65 degrees) . CONCLUSIONS: 1 . Vegetation dimension and mobility determined by TEE are important predictors of the embolic risk . 2 . Significant echocardiographic predictors of embolic events included vegetation length >15 mm, neck/thickness ratio >0.69, and maximal angle of displacement of vegetation during cardiac cycle >60.7 degrees . 3 . During antibiotic treatment, the embolic risk depends only on vegetation mobility and dimension.

J Bone Joint Surg Br, 2004 Aug, 86(6), 797 - 800
Microbiological culture results for the femoral head . Are they important to the donor?
James LA, Ibrahim T, Esler CN.
We determined the rate of contamination of donated femoral heads at primary hip arthroplasty within a single region between July 1992 and July 2001 . We established the null hypothesis that culture results played no role in predicting early failure of the joint because of infection . The rate of contamination was 9% . A positive culture, at the time of retrieval, was found in 367 of 4045 femoral heads . Coagulase-negative staphylococcus was isolated in 77% of the positive cases . At a minimum follow-up of one year, there was no statistically significant difference in the rate of complications or of revision of age-matched patients whose femoral heads had a positive culture compared with those whose femoral heads were sterile . Our findings confirm that culture of the femoral head plays no part in determining future failure of joint replacement in the donor.

Ann Pharmacother, 2004 Oct, 38(10), 1603 - 7 Epub 2004 Aug 24.
Effect of 24-hour intravenous tubing set change on the sterility of repackaged fat emulsion in neonates; Reiter PD et al.; BACKGROUND: Duration of intravenous fat emulsion (IVFE) infusions, precise method of administration (manufactured bottle vs repackaged syringe), and interval for administration set change continue to be debated . OBJECTIVE: To determine the contamination rate associated with replacing IVFE administration sets every 24 hours in newborn infants receiving fat emulsion repackaged into unit-of-use syringes . METHODS: This was a prospective, microbiologic study of 90 administration sets used in 19 neonates . IVFE samples were obtained from administration sets at the end of a 19- to 23-hour infusion and prior to daily tubing set change from infants who received repackaged IVFE . Samples of IVFE (1-3 mL) were aseptically removed at the catheter connection site proximal to the patient, transferred into BACTEC PEDSPlus culture media, and continuously monitored for 5 days to detect gram-positive and gram-negative organisms, as well as yeast . RESULTS: Two samples (2.27%) grew coagulase-negative Staphylococcus . Both samples were from the same asymptomatic patient and were obtained on consecutive days . A blood sample obtained through this infant's central catheter grew the same organism and suggested catheter hub colonization as the primary site of microbe origin . CONCLUSIONS: Microbial contamination of IVFE infusion sets changed at 24-hour intervals, using unit-of-use syringes in neonates, was low at 2.2%.

FEMS Immunol Med Microbiol, 2004 Sep 1, 42(1), 130 - 8
Interleukin-10 and sudden infant death syndrome; Moscovis SM et al.; Uncontrolled pro-inflammatory responses to infections or bacterial toxins have been suggested to play a role in triggering the physiological events leading to sudden infant death syndrome (SIDS) . We tested the hypothesis that these uncontrolled responses might be due to interactions between the gene polymorphisms inducing low levels of IL-10 and exposure to cigarette smoke . In vitro, the IL-10 (G-1082A) polymorphism was associated with low IL-10 levels and the -1082G allele was associated with high levels . The first objective was to assess the distribution of this polymorphism among SIDS infants, parents of SIDS infants and controls, and two ethnic groups: Aboriginal Australians who have a high incidence of SIDS; and Bangladeshis who in Britain have a low incidence of SIDS compared with Europeans . The second objective was to assess effects of human recombinant IL-10 on interleukin-6 (IL-6) and tumour necrosis factor-alpha (TNF-alpha) responses of human leukocytes to staphylococcal toxins implicated in SIDS . The third objective was to assess IL-10 responses to endotoxin and toxic shock syndrome toxin (TSST) from leukocytes of smokers and non-smokers in relation to the IL-10 (G-1082A) polymorphism . There were major differences in the distributions of these polymorphisms between Europeans and Bangladeshis (p=0.00) and between Europeans and Aboriginal Australians (p=0.00); however, they were similar for the Bangladeshi and Aboriginal Australian subjects . There were no significant differences in the distribution of these polymorphisms among SIDS infants or parents of SIDS infants compared to control groups . IL-10 significantly reduced IL-6 and TNF-alpha responses to TSST and staphylococcal enterotoxins A and C . At 50 ng ml(-1), IL-10 significantly increased TNF-alpha but not IL-6 responses to TSST and enterotoxin A . Although IL-10 responses to endotoxin were lower from leukocytes of smokers who were homozygous for the G allele, the differences were not significant; however, significantly lower IL-10 responses were found for smokers who were homozygous for the A allele (p=0.01) and heterozygotes (p=0.04) . The pooled data found smokers had significantly lower levels of IL-10 responses to TSST, but there were no significant differences for smokers compared with non-smokers for the three genotypes . The high incidence of SIDS and serious respiratory infections among Aboriginal Australian infants and the low incidence of these conditions among Bangladeshi infants might be explained in part by our findings of differences in IL-10 responses between smokers and non-smokers . The lowest levels of IL-10 responses were observed among smokers who were homozygous for the A allele which is most prevalent among the Aboriginal Australians (83%) and Bangladeshis (84%) . The major difference between the risk factors for SIDS in these two groups is the level of exposure of infants to cigarette smoke associated with maternal smoking.

Biosci Biotechnol Biochem, 2004 Aug, 68(8), 1663 - 71
Characterization of a gene cluster of Staphylococcus warneri ISK-1 encoding the biosynthesis of and immunity to the lantibiotic, nukacin ISK-1; Aso Y et al.; We characterized a gene cluster in a plasmid designated pPI-1 of Staphylococcus warneri ISK-1 encoding the biosynthesis of and immunity to the lacticin-481 type lantibiotic, nukacin ISK-1 . The DNA sequence suggested that the nukacin ISK-1 gene cluster consists of at least six genes, nukA (a structural gene), -M, -T, -F, -E, -G, and two open reading frames, ORF1 and ORF7 . NukM and NukT were predicted to be involved in post-translational modification and secretion of nukacin ISK-1 respectively . NukF, -E, and -G were predicted to form a membrane complex which contributes to self-protection from nukacin ISK-1 . Transcriptional analyses revealed that nukM through ORF7 comprises an operon, and that ORF1 is transcribed independently from downstream of nukA . The transcriptional levels of the nukA and nukM genes were enhanced by osmotic stress . The expression level of the nukA transcript was scarcely enhanced by nukacin ISK-1, suggesting that expression is not under the control of the autoregulatory circuit.

J Immunol, 2004 Sep 1, 173(5), 3201 - 8
Naive T cells are resistant to anergy induction by anti-CD3 antibodies; Andris F et al.; Anti-CD3 mAbs are potent immunosuppressive agents used in clinical transplantation . It has been generally assumed that one of the anti-CD3 mAb-mediated tolerance mechanisms is through the induction of naive T cell unresponsiveness, often referred to as anergy . We demonstrate in this study that naive T cells stimulated by anti-CD3 mAbs both in vivo and in vitro do not respond to the superantigen staphylococcal enterotoxin B nor to soluble forms of anti-CD3 mAbs and APC, but express increased reactivity to plastic-coated forms of the same anti-CD3 mAbs and to their nominal Ag/class II MHC, a finding that is difficult to rationalize with the concept of anergy . Phenotypic and detailed kinetic studies further suggest that a strong signal 1 delivered by anti-CD3 mAbs in the absence of costimulatory molecules does not lead to anergy, but rather induces naive T cells to change their mitogen responsiveness and acquire features of memory T cells . In marked contrast, Ag-experienced T cells are sensitive to anergy induction under the same experimental settings . Collectively, these studies demonstrate that exposure of naive T cells in vivo and in vitro to a strong TCR stimulus does not induce Ag unresponsiveness, indicating that sensitivity to negative signaling through TCR/CD3 triggering is developmentally regulated in CD4(+) T cells.

Infect Immun, 2004 Sep, 72(9), 5493 - 7
Development of a novel chloramphenicol resistance expression plasmid used for genetic complementation of a fliG deletion mutant in Treponema denticola; Slivienski-Gebhardt LL et al.; A new expression plasmid containing the fla operon promoter and a staphylococcal chloramphenicol resistance gene, was constructed to help assess the role of fliG in Treponema denticola motility . Deletion of fliG resulted in a nonmotile mutant with a markedly decreased number of flagellar filaments . Wild-type fliG genes from T . denticola and from Treponema pallidum were cloned into this expression plasmid . In both cases, the gene restored the ability of the mutant to gyrate its cell ends and enabled colony spreading in agarose . This shuttle plasmid enables high-level expression of genes in T . denticola and possesses an efficient selectable marker that provides a new tool for treponemal genetics.

Curr Protein Pept Sci, 2004 Aug, 5(4), 275 - 86
Linderstrom-Lang-Schellman's model for protein stabilization revisited; Taniuchi H et al.; The fact that cleavage of single peptide linkages in proteins often leads to extensive conformational alteration, including regions far removed from the cleavage site is not fully understood . We propose, based on the work of Linderstrom-Lang and Schellman, that disruption primarily occurs within protein structural domains that are stabilized by cooperative interactions and that cleavage of single peptide linkages of the domain perturbs the entire cooperative interaction . For this model we review experimental observations: on fragment complexation (ribonuclease A, staphylococcal nuclease and cytochrome c), destabilized N-terminal large fragments (ribonuclease A and nuclease), cooperative folding and stabilization of proteins (ribonuclease A, nuclease and cytochrome c), the close relationship of the three-dimensional structure between fragment complexes and the original protein (ribonuclease A and nuclease), ligand induced stabilization (nuclease), 3D domain swapping, circular permutation (dihydrofolate reductase), evolutionary conservation (cytochrome c fold) . Based on analysis of these observations, we conclude that the cooperative interactions of domains are important for the mechanism of 3D domain swapping as well as for stabilization and thereby, determination of the ground state of native proteins . Furthermore, analysis of the observations reveals that domains generally contain a hydrophobic core . Further, based on studies of cytochrome c and the Tsao, Evans and Wennerstrom model of electrostatic interactions between two hydrophobic monolayers, we propose the model that the hydrophobic core of a domain is polarizable and responds to the surface charges through its polarizability to stabilize the domain, explaining in part the nature of the cooperative interactions.

Presse Med, 2004 Jul 10, 33(12 Pt 2), 2S18 - 22
{Clinical efficacy and cost/benefit ratio of current treatment of MRSA infections in intensive care units}; Nathwani D; THE RISK FACTORS FOR BACTERIAL RESISTANCE: These are recent history of hospitalization and/or antibiotics, notably levofloxacine or macrolides, an increase in severity of the disease, prolonged hospitalization in intensive care before the diagnosis and the existence of decubitus bed sores . THE IMPACT OF MRSA INFECTIONS: Is clinical to start, with a greater mortality rate in MRSA bacteremias than in staphylococcal sensitive, in vancomycin-resistant enterococcal infections than in enterococcal sensitive infections, and in acquired pneumonia under mechanical ventilation . From an economical point of view, the progression in bacterial resistances in intensive care units has consequences not only on the patients and the hospital but also on the whole of society . TO FIGHT AGAINST NOSOCOMIAL INFECTIONS: Three measures should be taken: means of prevention must be used, appropriate antibiotic treatments must be administered early and days of hospitalization must be reduced.

Allergy Asthma Proc, 2004 May-Jun, 25(3), 181 - 4
Interleukin-18 is associated with increased severity of atopic dermatitis in children; Sohn MH et al.; Atopic dermatitis (AD) is a chronic relapsing skin disease characterized by reduced interferon (IFN) gamma production with concurrent up-regulation of interleukin (IL)-4 . Recently, it was reported that IL-18, formerly called IFN gamma-inducing factor, induces the production of T helper (Th)2-related cytokines without help from IL-12 . This study was performed to evaluate the contribution of IL-18 in the pathogenesis of AD . Significantly higher serum IL-18 concentrations were found in patients with severe AD than in healthy subjects . Under staphylococcal enterotoxin B stimulation, IL-18 secretion was increased in peripheral blood mononuclear cells from patients with AD . There were significant differences in the concentrations of IL-10, IL-12, and soluble Fas ligand between AD patients and normal controls . In conclusion, increased serum IL-18 concentrations may be involved in the pathogenesis of AD.

Amino Acids, 2004 Oct, 27(2), 175 - 81 Epub 2004 Aug 19.
Oxidation of buried cysteines is slow and an insignificant factor in the structural destabilization of staphylococcal nuclease caused by H(2)O(2) exposure; Kim YH et al.; The oxidation of buried cysteine or methionine residues can destroy the enzyme activity of a protein by disrupting structure . Engineering in such an oxidatively triggered switch for enzyme activity would only be useful if the effects of substitution are relatively minor, while the effects of the oxidized side chain upon structure are significant and the oxidation relatively easy . To assess the feasibility of this strategy for controlling enzyme activity, the effects of such substitutions and their oxidation were studied in a well characterized model protein, staphylococcal nuclease . Stability and enzyme activity of the oxidized proteins was assessed and compared to the stability and enzyme activity of the unoxidized proteins . Cysteines were found to be generally well tolerated in buried positions but these mutants were not more destabilized than wild-type when oxidized . This shows that buried cysteines are difficult enough to oxidize that this is not likely to be a useful protein engineering strategy or a commonly used regulatory modification . Similar effects were observed for methionine.

Probl Tuberk Bolezn Legk, 2004, (6), 11 - 7
{Staphylococcal pneumonia}; Dvoretskii LI et al.; In January to December 2000, a total of 15 (8.8%) patients with staphylococcal community-acquired pneumonia (CAP), 13 (27.7%) patients with staphyloccocal nosocomial pneumonia (NP) that occurred at general units (GU), and 9 (50%) patients with GU that occurred under artificial ventilation (AV) at intensive care units (ICU) were followed up . Meticillin-resistant S . aureus strains were isolated in 6.7, 38.5, and 55.6% of cases, respectively . As compared with pneumococcal CAP, staphyloccocal CAP were more frequently characterized by the severe course of the disease (46.7% versus 15.4%), bilateral lesion (33.3% versus 5.1%), and the presence of complications (66.7% versus 30.8%) . Staphyloccocal NP that had occurred at GU, as compared to that at ICU also showed the severe course of the disease (46.2% versus 2.9%), bilateral lesion (30.7% versus 0%), and developed complications (75% versus 25%) . Staphylococcal NP developed under AV at ICU had no specific features as compared with NP of another etiology . Oxacillin and first-second-generation cephalosporins remain to be the drugs of choice when meticillin-sensitive S . aureus strains are isolated; lincomycin and erythromycin being alternative agents against these strains . Glycopeptides are the drugs of choice when meticillin-resistant S . aureus strains are isolated, its alernatives are linesolide or rifampicin . High mortality rates due to staphylococcal pneumonia are preserved . These are 7.1% in CAP, 7.7 and 66.7 in staphylococcal NP occurring at GU and under AV at ICU, respectively.

Int Immunopharmacol, 2004 Oct, 4(10-11), 1343 - 51
Differential regulation of Th1/Th2 in relevant tissues for sepsis pathogenesis with a Limulus anti-LPS factor-derived peptide increases survival in Gram-positive sepsis; Vallespi MG et al.; Severe sepsis and septic shock are important causes of death in intensive care units . Although Gram-negative infections were predominant in the 1960s, Gram-positive infections have increased in the past two decades and now account for about half of the cases of severe sepsis . In this study, we examined the effect of a Limulus anti-LPS factor (LALF)-derived peptide on lung and liver Th1/Th2 cytokine mRNA levels during a Gram-positive sepsis . We also examined the morphopathological changes observed in these organs during the disease . Mice challenged with a high dose of Staphylococcus haemolyticus showed severe damage in lung . In contrast, the liver of challenged mice showed an accumulation of bacterial particles in the sinusoids, associated with a severe inflammatory response due to high levels of tissue mRNA proinflammatory cytokines . Treatment with the peptide LALF(32-51) ameliorated the sepsis-induced effects in the lung and liver and increased the survival of mice in a dose- and time-dependent manner . Pretreatment with the peptide LALF(32-51) differentially regulates TNF-alpha, IFN-gamma, IL-12p40, IL-2 and IL-10 mRNA levels in lung and liver of peptide-treated mice, and limits the systemic inflammatory response . These findings support for the first time the effectiveness of an LALF-derived peptide in the treatment of a Gram-positive sepsis . Modulation of the Th1/Th2 pattern in tissues relevant for sepsis correlates with an improved outcome of the disease as denoted by increased survival.

J Cataract Refract Surg, 2004 Aug, 30(8), 1802 - 4
Keratitis after implantation of intrastromal corneal ring segments; Shehadeh-Masha'our R et al.; A 53-year-old man had implantation of 2 intrastromal corneal ring segments (Intacs, Addition Technology Inc.) in the left eye . Several days after the lower segment was exchanged, an infiltrate was present at the corneal incision site . This progressed to a lower channel infection followed by diffuse keratitis . Culture from the conjunctiva was positive for Staphylococcus epidermidis, but cultures from the cornea were negative for microorganisms . Later, 2 gas bubbles appeared at the interface . The patient was treated with topical and subconjunctival antibiotics and channel irrigation . The keratitis resolved, leaving a neovascularized opacity at the nasal part of the lower channel.

Rocz Panstw Zakl Hig, 2004, 55(1), 89 - 98
{Assessment of the probability of encountering staphylococcal enterotoxins in lactic acid cheese packaged in laminates}; Steinka I; Immunoassay methods were used to identify the presence of staphylococcal enterotoxins in lactic acid cheese vacuum and non-vacuum packed . There was assessed the probability of encountering staphylococcal enterotoxin in cheese dependent on different systems of packaging, count of staphylococcal cells, intensiveness of coagulase synthesis and tightness of packaging . The presence of enterotoxin was identified in 5% of researched samples of products stored for 14 days . The influence of packaging system and tightness on presence of enterotoxin was observed . The probability of presence of staphylococcal and enterotoxin in relation to researched factors was presented by the mathematical models.

Clin Microbiol Infect, 2004 Aug, 10(8), 765 - 8
Penetration of fusidic acid and rifampicin into cerebrospinal fluid in low-grade inflammatory meningitis caused by Staphylococcus epidermidis; Hedberg A et al.; Cerebrospinal fluid (CSF) concentration-time curves of rifampicin and fusidic acid were studied in a patient with post-operative meningitis caused by Staphylococcus epidermidis . The patient was treated with this combination of antimicrobial agents because of a severe hypersensitivity reaction to vancomycin . Peak CSF concentrations of rifampicin exceeded the MIC by > 60-fold, while those of fusidic acid just reached the MIC . CSF concentrations of fusidic acid were relatively stable within the range reported for patients with uninflamed meninges, but serum levels were surprisingly low . An increase in the metabolism of fusidic acid induced by rifampicin cannot be excluded.

Acta Crystallogr D Biol Crystallogr, 1995 Nov, 51(Pt 6), 1080 - 1
Preliminary X-ray studies on two new crystal forms of staphylococcal enterotoxin C2; Swaminathan S; Two new crystral forms of staphylococcal enterotoxin C2 have been obtained by vapor-diffusion methods . Form 1 crystals are monoclinic in space group P2(1) with cell dimensions a = 43.43, b = 69.92, c = 42.22 A, 8 = 90.1 degrees and diffract to at least 2.7 A resolution . Form 2 crystals are tetragonal in space group P4(1)2(1)2 or P4(3)2(1)2 with cell dimensions a = b = 42.98, c = 289.92 A and diffract to 1.9 A resolution.

Echocardiography, 2004 Aug, 21(6), 555 - 8
Precordial abscess inducing chest pain 20 years after surgical repair of a pentalogy of fallot; Aschwanden E et al.; A 25-year-old male asylum-seeker presented with chest pain, exertional dyspnea, and orthopnea 20 years after the surgical repair of a pentalogy of Fallot . An extracardiac mass compressing the right ventricle was subsequently detected and surgical decompression was performed to relieve the resulting right intraventricular hypertension . At operation, the mass proved to be a coagulase-negative, staphylococcal abscess . In addition, the removal of the mass unmasked a previously nonrecognized pulmonary outflow stenosis that required balloon dilatation and beta-blocker therapy . While infections are known to occur after sternotomy, the formation of an abscess in the anterior mediastinum several years after the intervention appears to be exceptional; this diagnosis came to mind only after the more common complications had been considered, e.g., pseudoaneurysm or pericardial hematoma . To our knowledge, this is the first report of an abscess in the anterior mediastinum that had probably formed over many years following a sternotomy, compressed the right ventricle and masked a pulmonary stenosis.

Clin Exp Allergy, 2004 Aug, 34(8), 1270 - 5
Specific immunoglobulin E for staphylococcal enterotoxins in nasal polyps from patients with aspirin-intolerant asthma; Suh YJ et al.; BACKGROUND: Nasal polyps infiltrated with eosinophils are commonly found in chronic asthmatic patients, more frequently in those with aspirin-intolerant asthma (AIA) than aspirin-tolerant asthma (ATA) . Some studies have suggested a contribution of superantigens derived from Staphylococcus sp to nasal polyposis and eosinophilia, but their relative importance in AIA and ATA subjects is unknown . OBJECTIVE: We investigated whether local production of specific IgE to staphylococcal enterotoxins A and B (SEA and SEB) and relationships with markers of eosinophilic inflammation differ in the nasal polyps of AIA and ATA subjects . METHODS: Fifteen AIA subjects with positive responses to lysine-aspirin bronchoprovocation and 15 ATA subjects underwent polypectomy . Immunoassays were used to quantify eosinophil cationic protein (ECP), IL-5, mast cell tryptase, soluble IL-2 receptors (sIL-2R), total IgE, and specific IgE for SEA and SEB . RESULTS: ECP levels in nasal polyp homogenates were higher in AIA subjects than in ATA subjects (P < 0.02), with no significant differences in tryptase, IL-5 or sIL-2R . Total IgE, and specific IgE to both SEA and SEB, were detectable in some nasal polyps from both subject groups, but median levels were markedly higher in AIA subjects than in ATA subjects (P = 0.04, 0.01, 0.05, respectively) . Levels of specific IgE to SEA and SEB correlated significantly with levels of ECP and IL-5, but not those of tryptase or sIL-2R . CONCLUSION: These findings suggest that staphylococcal superantigens may drive local eosinophilic inflammation in nasal polyp tissue, and that this is exacerbated in subjects with AIA.

J Clin Microbiol, 2004 Aug, 42(8), 3549 - 57
Use of 16S ribosomal DNA PCR and denaturing gradient gel electrophoresis for analysis of the microfloras of healing and nonhealing chronic venous leg ulcers; Davies CE et al.; The bacterial microfloras of 8 healing and 10 nonhealing chronic venous leg ulcers were compared by using a combination of cultural analysis and denaturing gradient gel electrophoresis (DGGE) of PCR-amplified 16S rRNA gene products . Cultural analysis of the microflora revealed that the majority of both wound types carried the aerobes Staphylococcus and Pseudomonas spp . (89 and 80%, respectively) . Sequencing of 16S ribosomal DNAs selected on the basis of DGGE profiling allowed the identification of strains not detected by cultural means . Of considerable interest was the finding that more than 40% of the sequences represented organisms not cultured from the wound from which they were amplified . DGGE profiles also revealed that all of the wounds possessed one apparently common band, identified by sequencing as Pseudomonas sp . The intensity of this PCR signal suggested that the bacterial load of nonhealing wounds was much higher for pseudomonads compared to healing wounds and that it may have been significantly underestimated by cultural analysis . Hence, the present study shows that DGGE could give valuable additional information about chronic wound microflora that is not apparent from cultural analysis alone.

Acta Biochim Biophys Sin (Shanghai), 2004 Aug, 36(8), 577 - 82
Development of cell lines stably expressing staphylococcal nuclease fused to dengue 2 virus capsid protein for CTVI; Qin CF et al.; To explore the potential application of capsid-targeted viral inactivation (CTVI) strategy in prophylactic model against dengue virus (DV) infection, here we fused a Ca(2+)-dependent nuclease, staphylococcal nuclease (SN), to the capsid protein of dengue 2 virus (D2C) at the carboxyl terminal, and constructed the desired expression plasmid pc/D2C-SN and control plasmids pc/D2C-SN* and pc/D2C . A mammalian cell line BHK-21 was transfected by electroporation with those plasmids and thereafter selected by 5mgr;/ml blasticidin . The resistant cell clones were then expanding cultured and screened by RT-PCR and Western Blot assays . The nuclease activity of the expressed fusion protein D2C-SN was analyzed by in vitro DNA digestion assay . It was confirmed cell lines stably expressing D2C-SN and control constructs were obtained . The intracellular expressed fusion protein D2C-SN had ideal nuclease activity and no cytotoxicity on mammalian cells . Those engineered cell lines provided the experimental system for CTVI application in prophylactic model and paved the new road for combating DV infection with CTVI.

Acta Orthop Belg, 2004 Jun, 70(3), 290 - 4
Septic arthritis of a lumbar facet joint case report and review of the literature; Smida M et al.; Septic arthritis of a lumbar facet joint (SALFJ) is a very rare condition . It has mostly been described in adults . Only one other paediatric case has been reported . We present a case of septic arthritis of the left L5-S1 lumbar facet joint, associated with epiduritis and paraspinal abscess, in an 8-year-old boy . Plain radiographs and Technetium bone scan were negative . The diagnosis was made by blood cultures, which isolated staphylococcus epidermidis, and by MRI . The child was treated successfully with antibiotics only.

J Nutr, 2004 Aug, 134(8 Suppl), 2056S - 2061S
Canine atopic dermatitis: new targets, new therapies; DeBoer DJ; Atopic dermatitis is a common allergic skin disease of complex etiopathogenesis in both humans and dogs . Immediate-type hypersensitivity to environmental allergens that arises as a result of environmental and genetic factors is a major part of the pathogenesis in most but not all patients . Alterations in epidermal barrier function, priming of cutaneous antigen-presenting cells with IgE, intrinsic keratinocyte defects, and even development of autoimmunity are also factors that contribute to the primary disease . Secondary factors, especially infections with Staphylococcus and yeast organisms, strongly influence the course of this skin disease . The relatively recent understanding of the complexities of atopic dermatitis has resulted in changes in diagnostic and therapeutic strategies for the disease . We now know that the best therapeutic approach is to use combinations of multiple modalities individualized for each patient over the course of his or her lifetime.

J Mol Biol, 2004 Aug 6, 341(2), 565 - 74
X-ray and thermodynamic studies of staphylococcal nuclease variants I92E and I92K: insights into polarity of the protein interior; Nguyen DM et al.; We have used crystallography and thermodynamic analysis to study nuclease variants I92E and I92K, in which an ionizable side-chain is placed in the hydrophobic core of nuclease . We find that the energetic cost of burying ionizable groups is rather modest . The X-ray determinations show water molecules solvating the buried glutamic acid under cryo conditions, but not at room temperature . The lysine side-chain does not appear solvated in either case . Guanidine hydrochloride (GnHCl) denaturation of I92E and I92K, done as a function of pH and monitored by tryptophan fluorescence, showed that I92E and I92K are folded in the pH range pH 3.5-9.0 and pH 5.5-9.5, respectively . The stability of the parental protein is independent of pH over a broad range . In contrast, the stabilities of I92E and I92K exhibit a pH dependence, which is quantitatively explained by thermodynamic analysis: the PK(a) value of the buried K92 is 5.6, while that of the buried E92 is 8.65 . The free energy difference between burying the uncharged and charged forms of the groups is modest, about 6 kcal/mol . We also found that epsilon(app) for I92K and I92E is in the range approximately 10-12, instead of 2-4 commonly used to represent the protein interior . Side-chains 92E and 92K were uncharged under the conditions of the X-ray experiment . Both are buried completely inside the well-defined hydrophobic core of the variant proteins without forming salt-bridges or hydrogen bonds to other functional groups of the proteins . Under cryo conditions 92E shows a chain of four water molecules, which hydrate one oxygen atom of the carboxyl group of the glutamic acid . Two other water molecules, which are present in the wild-type at all temperatures, are also connected to the water ring observed inside the hydrophobic core . The ready burial of water with an uncharged E92 raises the possibility that solvent excursions into the interior also take place in the wild-type protein, but in a random, dynamic way not detectable by crystallography . Such transient excursions could increase the average polarity, and thus epsilon(app), of the protein interior.

Ther Apher Dial, 2004 Aug, 8(4), 281 - 5
Immunoadsorption in lupus myocarditis; Griveas I et al.; Reduction of pathological autoantibodies may be useful in the treatment of systemic lupus erythematosus (SLE) . On the other hand clinically manifested myocarditis in SLE, though uncommon, may be life-threatening and its pathogenesis has been ascribed to autoimmunity . The aim of this study is to present a rare case of a patient with severe lupus myocarditis, where immunoadsorption (IA) was evaluated as rescue therapy . A case of SLE with initial manifestation of myocarditis is reported in a 29-year-old male who presented with arthritis, fever, lymphadenopathy, joint swelling and morning stiffness . Laboratory evaluation revealed increased antinuclear antibody (ANA), slightly decreased complement and positive anticoagulant panel . From the above clinical and laboratory features, criteria of SLE seemed applicable . During his hospitalization, the patient developed pericardial effusion and cardiogenic shock . Although pericardiotomy was performed and was treated with immunosuppressive agents, plasmapheresis and supported with current medications, his clinical condition remained critical with an ejection fraction of 20% . At this point it was decided to receive IA onto staphylococcal protein A . After 6 sessions with IA and concomitant immunosuppression, the patient responded well, his condition improved and was dismissed with an ejection fraction of 50% . Fulminant lupus myocarditis is a severe and rare situation lacking a satisfying specific therapy available today . In our presented case, IA in addition to immunosuppressive therapy was beneficial . Considering the benefits of our case and the current knowledge, it might be useful to clarify the open question in scale pilot studies.

Bull Exp Biol Med, 2004 Feb, 137(2), 175 - 8
Effects of secretory products of activated neutrophils on morphological composition and functional activity of peritoneal exudation cells during inflammation of staphylococcal origin; Tretyakova IE et al.; Substance A5 isolated from supernatants of activated neutrophils from donors significantly increases the percentage of neutrophils and macrophages in the peritoneal exudation of mice on days 3 and 7 of staphylococcal inflammation and stimulates functional activity (lysosomal, phagocytic, and NBT-reducing) of these cells, reduced as a result of inflammation, on days 3, 7, and 14 of the inflammatory process.

Antimicrob Agents Chemother, 2004 Aug, 48(8), 3162 - 4
Temporin A soaking in combination with intraperitoneal linezolid prevents vascular graft infection in a subcutaneous rat pouch model of infection with Staphylococcus epidermidis with intermediate resistance to glycopeptides; Giacometti A et al.; The efficacy of linezolid and temporin A in the prevention of prosthetic graft infection due to methicillin-resistant Staphylococcus epidermidis with intermediate resistance to glycopeptides was investigated in a subcutaneous rat pouch model . Linezolid and temporin A, alone or combined, greatly reduced the bacterial numbers compared to the effect with control drugs.

FEMS Microbiol Lett, 2004 Aug 1, 237(1), 157 - 61
Cloning of an agr homologue of Staphylococcus saprophyticus; Sakinc T et al.; An agr homologue of Staphylococcus saprophyticus was identified, cloned and sequenced . The gene locus shows homologies to other staphylococcal agr systems, especially to those of S . epidermidis and S . lugdunensis . A putative RNAIII was identified and found to be differentially expressed during the growth phases . In contrast to the RNAIII molecules of S . epidermidis and S . aureus it does not contain an open reading frame that codes for a protein with homologies to the delta-toxin . Using PCR, the agr was found to be present in clinical isolates of S . saprophyticus.

Brain Behav Immun, 2004 Sep, 18(5), 476 - 84
The opioid antagonist naltrexone blocks acute endotoxic shock by inhibiting tumor necrosis factor-alpha production; Greeneltch KM et al.; Septic shock is believed to be a consequence of excessive stimulation of the immune system by bacterial toxins that results in systemic overproduction of proinflammatory cytokines, including tumor necrosis factor-alpha (TNF-alpha), IL-1, and IL-6 . Various studies have shown that TNF-alpha, a major mediator of septic shock, induces tissue injury, loss of blood pressure, organ failure, and ultimately death . Administration of the opioid antagonist naloxone has been reported to reverse opiate-mediated hypotension, promote organ perfusion and increase patient survival . In this study, we examined the mechanism by which the opioid receptor antagonist, naltrexone, modulates the septic shock response in BALB/c mice after injection with lipopolysaccharide (LPS) or staphylococcal enterotoxin B (SEB) in combination with d-galactosamine (d-gal), or with agonistic anti-Fas antibody (Jo2) alone . Each of these treatments induced rapid-onset, acute shock, and ultimately mortality (6-9h after injection), although different mechanisms are involved . Administration of the opioid antagonist naltrexone protected mice from shock induced by LPS+d-gal, but not SEB+d-gal or Jo2 antibody, a protective effect that was reversed by morphine . Naltrexone significantly inhibited the production of TNF-alpha induced by LPS, but not SEB in vivo . When bone marrow-derived, splenic or peritoneal macrophages were treated with LPS in vitro, administration of naltrexone had no direct effect on TNF-alpha production . These results suggest that naltrexone is capable of preventing LPS-induced septic shock mortality by indirect inhibition of TNF-alpha production in vivo.

Curr Infect Dis Rep, 2004 Aug, 6(4), 263 - 269
Intra-arterial Device Infections; Antonios VS et al.; As an increasing number and variety of prosthetic devices are used in cardiovascular medicine, novel infectious complications have been described . Infection of intra-arterial devices, including arterial closure devices, prosthetic carotid patches, coronary artery stents and endovascular stents, and stent-grafts, is now being reported . Prosthetic vascular graft infection is an older, more common, and better-characterized entity, but recent developments in the surgical management of these infections have prompted a re-examination of the syndrome . Staphylococcal species account for most intra-arterial device infections, and often, morbidity and mortality rates are high . An update on intra-arterial device infections is warranted.

J Biol Chem, 2004 Sep 24, 279(39), 41028 - 37 Epub 2004 Jul 18.
Crystal structure of leucotoxin S component: new insight into the Staphylococcal beta-barrel pore-forming toxins; Guillet V et al.; Staphylococcal leucocidins and gamma-hemolysins (leucotoxins) are bi-component toxins that form lytic transmembrane pores . Their cytotoxic activities require the synergistic association of a class S component and a class F component, produced as water-soluble monomers that form hetero-oligomeric membrane-associated complexes . Strains that produce the Panton-Valentine leucocidin are clinically associated with cutaneous lesions and community-acquired pneumonia . In a previous study, we determined the crystal structure of the F monomer from the Panton-Valentine leucocidin . To derive information on the second component of the leucotoxins, the x-ray structure of the S protein from the Panton-Valentine leucocidin was solved to 2.0 angstrom resolution using a tetragonal crystal form that contains eight molecules in the asymmetric unit . The structure demonstrates the different conformation of the domain involved in membrane contacts and illustrates sequence and tertiary structure variabilities of the pore-forming leucotoxins . Mutagenesis studies at a key surface residue (Thr-28) further support the important role played by these microheterogeneities for the assembly of the bipartite leucotoxins .

Curr Eye Res, 2004 Apr, 28(4), 225 - 32
Intravitreal taurolidine against experimental Staphylococcus epidermidis endophthalmitis in rabbits; Oguz H et al.; PURPOSE: Taurolidine is a broad spectrum, non-antibiotic antimicrobial agent, not previously tested against infectious endophthalmitis . The efficacy of intravitreal taurolidine in the treatment of experimental Staphylococcus epidermidis endophthalmitis was evaluated and compared with vancomycin in a rabbit model . METHODS: The right eyes of 34 albino rabbits were infected with an intravitreal inoculum of S . epidermidis (10(5) colony-forming units/0.1 ml) . The right eyes of four rabbits (group 7) were not infected and served as uninfected controls . 24 hours after inoculation of bacteria the animals were divided into the following treatment groups: group 1 (7 rabbits) received intravitreal taurolidine at 24 hours and group 2 (7 rabbits) received at 48 hours . Group 3 (7 rabbits) received vancomycin at 24 hours and group 4 (7 rabbits) at 48 hours . Group 5 (3 rabbits) received polyvinylpyrrolidone at 24 hours and group 6 (3 rabbits) at 48 hours . Clinical scoring was performed at 24, 48 and 72 hours . At 72 hours post inoculation, vitreous samples were collected for quantitative microbiological studies and then, the eyes were enucleated for histopathological scorings . RESULTS: The clinical and histopathological examinations revealed significant amelioration of inflammation in eyes treated with taurolidine and vancomycin when compared with polyvinylpyrrolidone . The eyes treated with taurolidine also had significantly lower colony forming units than the eyes treated with polyvinylpyrrolidone and taurolidine rendered many eyes sterile . CONCLUSION: Taurolidine is expected to be a potential agent for treatment of S . epidermidis endophthalmitis.

Biol Chem, 2004 Jun, 385(6), 543 - 6
Characterisation of a highly specific, endogenous inhibitor of cysteine protease from Staphylococcus epidermidis, a new member of the staphostatin family; Dubin G et al.; Staphostatins, a novel family of cysteine protease inhibitors with a unique mechanism of action and distinct protein fold has recently been discovered . In this report we describe the properties of Staphylococcus epidermidis staphostatin A (EcpB), a new member of the family . As for other staphostatins, the recombinant S . epidermidis staphostatin A exerted very narrow inhibitory specificity, limited to cysteine protease from the same species . The closely related proteases from S . aureus cleaved the inhibitor at the reactive site peptide bond and inactivated it . The EcpB homologue, S . aureus staphostatin A (ScpB), was also susceptible to proteolytic cleavage at the same site by non-target cysteine proteases . Conversely, S . aureus staphostatin B (SspC) was resistant to such proteolysis . The difference in the susceptibility of individual inhibitors to proteolytic cleavage at the reactive site suggests subtle variations in the mechanism of interaction with cysteine proteases.

Int J Mol Med, 2004 Aug, 14(2), 241 - 6
Role of caspase in CD80 expression of superantigen-stimulated monocytes; Shinohara F et al.; In this study, we demonstrated evidence for the induction of CD80+ monocytes by staphylococcal enterotoxin B (SEB) via caspase actions . Pre-treatment with pyrrolidine dithiocarbamate (PDTC), a potent inhibitor of NF-kappaB, resulted in a significant reduction in the percentage of SEB- and interferon-gamma (IFN-gamma) (produced by SEB) -induced CD80+ monocytes . SEB and IFN-gamma activated NF-kappaB, which was inhibited by PDTC . SEB induced the activation of caspase-3 and -8, and pre-treatment with z-VAD-fmk, a broad-spectrum inhibitor of caspases, prevented the induction of apoptosis . Treating with z-VAD-fmk resulted in a reduction of the generation of CD80+ monocytes . These results indicated that CD80 driven by NF-kappaB is regulated by the enzymatic actions of caspases, which allows monocytes to participate in massive T-cell activation.

J Altern Complement Med, 2004 Jun, 10(3), 519 - 26
Yin zi huang, an injectable multicomponent chinese herbal medicine, is a potent inhibitor of T-cell activation; Chen X et al.; OBJECTIVES: The clinical efficacy of many multiherbal Traditional Chinese Medicines (TCM) is partially attributable to their immunoregulatory properties . In this study we evaluated the effect of eight commonly used, commercially available multiherbal Chinese medicines on T-cell activation . We focused on Yin Zhi Huang (YZH, an injectable herbal medicine commonly used for the treatment of liver diseases in China), because it was the most potent inhibitor of T-cell activation in our experimental system . The effects of 10 ingredient components of YZH were also evaluated . METHODS: {3H} thymidine incorporation assay was used to assess mouse T-cell proliferation after stimulation with latex beads coated with anti-CD3/CD28 antibodies . CD25, CD69, PD-1, and I-COS expression by purified mouse CD4+ T cells treated with plate-bound anti-CD3 antibody and soluble anti-CD28 antibody was analyzed by fluorescent-activated cell sorter (FACS) . Cytokine/chemokine production by human peripheral blood mononuclear cells (PBMC) stimulated with staphylococcal enterotoxin B (SEB) was determined by enzyme-linked immunosorbent assay (ELISA) . RESULTS: Among tested herbal medicines, YZH was the most potent inhibitor of T-cell activation . In splenocyte proliferation assays, the inhibitory effect of YZH was dose-dependent, with a 50% inhibition concentration (IC50) of 1:3200-1:1600 . Ten (10) purified compounds found in YZH were evaluated for their activity . Among them, ursolic acid (1-10 micromol), luteolin (1-10 micromol), baicalein (1-10 micromol), scopran (5-50 micromol), and crocin (5-50 micromol), exhibited dose-dependent inhibition . YZH also inhibited CD25, CD69, PD-1, and ICOS expression by stimulated mouse CD4+ T cells . In human PBMCs, YZH inhibited SEB-stimulated cytokine (interleukin {IL}-1, IL-2, IL-6, tumor necrosis factor{TNF}-alpha, interferon {IFN}-gamma) and chemokine (IP-10, MCP-1, MIP-1alpha and MIP-1beta) production in a dose-dependent manner . CONCLUSION: Our data show for the first time that YZH is a potent inhibitor of T-cell activation, and this property may be the major mechanism underlying the clinical efficacy of YZH . Our experimental results pave the way for identification of active component(s) and/or analysis of synergistic/additive effect of a YZH ingredient in future studies . Copyright Mary Ann Liebert, Inc.

Kidney Int, 2004 Aug, 66(2), 801 - 5
Locking of tunneled hemodialysis catheters with gentamicin and heparin; McIntyre CW et al.; INTRODUCTION: Catheter-related infection (CRI) is a major cause of morbidity and mortality in patients receiving hemodialysis . Antibiotic locking of these catheters has been shown to increase both the success of systemic antibiotic treatment in line sepsis, and to reduce the incidence of sepsis . We have studied the use of gentamicin locking of catheters (in combination with standard heparin rather than previously reported citrate) to reduce CRI rates . Furthermore, we have investigated the effects of this strategy on epoetin requirements and vascular access function . METHODS: Fifty patients were studied . Patients were randomized to catheter-restricted filling with either standard heparin (5000 IU/mL) alone, or gentamicin and heparin (5 mg/mL) . Epoetin requirements and hemoglobin response were monitored over the study period . RESULTS: The gentamicin-locked group suffered only one infective episode (0.3/1000 catheter days) compared to 10 episodes in six patients in the heparin alone group (4/1000 catheter days, P= 0.02) . The isolated organisms were equally split between Staphylococcal species and coliforms . There were no statistically significant differences in delivered dialysis dose (Kt/V) or QA between the two groups . Use of antibiotic locking was associated with both a higher mean hemoglobin (10.1 +/-0.14 g/dL vs . 9.2 +/- 0.17 g/dL in the heparin group, P= 0.003) and a lower mean epoetin dose (9000 +/- 734 IU/week vs . 10790 +/-615 IU/week in the heparin group, P= 0.04) . CONCLUSION: The practice of locking newly inserted tunneled central venous catheters with gentamicin and heparin is an effective strategy to reduce line sepsis rates, and is associated with beneficial effects on epoetin requirements.

Endocr Pract, 1998, 4(3), 159 - 64
Transient ionized hypocalcemia and secondary hyperparathyroidism accompanying acute adrenal insufficiency; Taylor MD FACP FACE HC et al.; Objective: To describe the hitherto unrecognized occurrence of transient ionized hypocalcemia with acute adrenal insufficiency and its therapy . Methods: We present three case reports with documented longitudinal laboratory findings . Results: Transient ionized hypocalcemia of acute illness has been noted in children and adults and is associated with increased mortality . Precipitating illnesses include gram-positive and gram-negative sepsis and staphylococcal toxic shock syndrome . We encountered three patients with transient ionized hypocalcemia associated with acute adrenal insufficiency . Similar to severely ill, transiently hypocalcemic patients without adrenal insufficiency, one patient demonstrated 25-hydroxyvitamin D deficiency, a second had minimal magnesium deficiency, and a third had no identifiable underlying abnormality . All three patients exhibited a transient increase in levels of serum intact parathyroid hormone and 1,25-dihydroxyvitamin D in response to ionized hypocalcemia, indicative of temporary secondary hyperparathyroidism . Two of the three patients were treated solely with glucocorticoids and intravenous administration of fluids, whereas the third received minimal intramuscularly administered magnesium and antibiotics in addition . All ultimately demonstrated a return to normal of serum total and ionized calcium, parathyroid hormone, and 1,25-dihydroxyvitamin D with no further treatment, even though one patient remained deficient in 25-hydroxyvitamin D . Conclusion: On the basis of these cases, we conclude that acute adrenal insufficiency and its treatment must be added to the disorders associated with transient ionized hypocalcemia and that transient secondary hyperparathyroidism is characteristic of at least some of the patients.

J Biomater Appl, 2004 Jul, 19(1), 47 - 57
In vitro attachment of Staphylococcus epidermidis to surgical sutures with and without Ag-containing bioactive glass coating; Pratten J et al.; The ability of a silver-doped bioactive glass (AgBG) coating to prevent bacterial colonization on surgical sutures was investigated in vitro . Bioactive glass powders, in the form of 45S5 Bioglass and AgBG, were used to coat Mersilk sutures using an optimized 'in house' slurry-dipping process . In vitro experiments were carried out using Staphylococcus epidermidis under both batch and flow conditions . While the traditional batch culture testing was used to determine the number of viable cells adhered to the surface, the flow-cell was used to visualize attachment and detachment over time . Under batch conditions of up to 180 min, statistically significant differences were observed in the colony forming units (CFU) per suture for both the coated and uncoated Mersilk sutures . The results showed that the AgBG coating had the greatest effect on limiting bacterial attachment (8 x 10(2) CFU) when compared to the 45S5 Bioglass coating (3.2 x 10(3) CFU) and the uncoated Mersilk (1.2 x 10(4) CFU) . Also under flow conditions differences were seen between the coated and uncoated sutures . Therefore, this preliminary study has demonstrated the quantification and visualization of bacterial attachment onto sutures in order to compare the antibacterial properties of Ag-containing bioactive glass coatings . The bactericidal properties imparted by Ag-containing glass open new opportunities for use of the composite sutures in wound healing and body wall repair.

Infect Control Hosp Epidemiol, 2004 Jun, 25(6), 498 - 503
A protracted outbreak of Staphylococcus epidermidis infections among patients undergoing valve replacement; Bou R et al.; OBJECTIVE: To investigate a Staphylococcus epidermidis outbreak among patients undergoing cardiac surgery . DESIGN: Retrospective cohort study . SETTING: A 260-bed community referral center . PATIENTS: Case-patients were patients with S . epidermidis mediastinitis, endocarditis, or both after valve implantation at Hospital de La Ribera from January to June 2002 . The study population included patients undergoing valve surgery at Hospital de La Ribera from January 2000 to June 2002 . RESULTS: From January to June 2002, 8 cases of mediastinitis, endocarditis, or both occurred among 53 patients undergoing cardiac surgery . In the same months of 2000, there had been no cases among 22 patients, and in 2001, only 1 case among 47 patients (P = .095 and P = .034, respectively) . In 2002, there were 4 cases of mediastinitis and endocarditis, 3 cases of mediastinitis, and 1 case of endocarditis, all following aortic valve replacement . The epidemic curve suggested a protracted outbreak . Patients with chronic obstructive lung disease were six-fold more likely to be case-patients (95% confidence interval, 1.6-23.8) . The mean duration of surgery tended to be longer in non-case-patients (161.4 +/- 57.9 minutes) than in case-patients (123.7 +/- 23.7 minutes) (P = .06) . CONCLUSION: The cause of this protracted outbreak was likely multifactorial . Reemphasis of existing policies was associated with resolution of the outbreak.

Chang Gung Med J, 2004 Apr, 27(4), 268 - 74
Nonunion of the femur treated with conventional osteosynthesis combined with autogenous and strut allogeneic bone grafts; Weng LH et al.; BACKGROUND: In this study, we reviewed the results of conventional osteosynthesis combined with strut onlay allografts and autogenous bone grafts as the treatment for nonunion of the femur . METHODS: We conducted a retrospective study of 18 patients with nonunion of the femur treated with internal fixation (eight 95 degree fixed-angled blade plates, 7 intramedullary nails, 2 dynamic hip compression plates and screws, and 1 condylar buttress plate) and autogenous bone grafts and cortical strut allografts . There were 1.8 operations on average before surgery . The average time from the initial treatment of the fracture or osteotomy to surgery was 32.5 months . The average follow-up was 32.2 months . RESULTS: All 18 nonunions had healed by the time of follow-up . Positive intraoperative cultures (Staphylococcus epidermidis) in 2 patients were resolved by parenteral antibiotics with no additional treatment . Removal of implants was required in 5 patients because of screw irritation or bony union . One concomitant quadricepsplasty as well as 2 excisions of a protruding graft were required because of restricted knee motion or impingement . CONCLUSIONS: For difficult nonunions of the femur in the current study, strict adherence to the principles of the treatment of nonunion and the addition of strut allografts to enhance stability and repair potential proved to be a good alternative.

Protein Sci, 2004 Aug, 13(8), 2078 - 88 Epub 2004 Jul 06.
Biophysical characterization of Z(SPA-1)--a phage-display selected binder to protein A; Lendel C et al.; Affibodies are a novel class of binding proteins selected from phagemid libraries of the Z domain from staphylococcal protein A . The Z(SPA-1) affibody was selected as a binder to protein A, and it binds the parental Z domain with micromolar affinity . In earlier work we determined the structure of the Z:Z(SPA-1) complex and noted that Z(SPA-1) in the free state exhibits several properties characteristic of a molten globule . Here we present a more detailed biophysical investigation of Z(SPA-1) and four Z(SPA-1) mutants with the objective to understand these properties . The characterization includes thermal and chemical denaturation profiles, ANS binding assays, size exclusion chromatography, isothermal titration calorimetry, and an investigation of structure and dynamics by NMR . The NMR characterization of Z(SPA-1) was facilitated by the finding that trimethylamine N-oxide (TMAO) stabilizes the molten globule conformation in favor of the fully unfolded state . All data taken together lead us to conclude the following: (1) The topology of the molten globule conformation of free Z(SPA-1) is similar to that of the fully folded structure in the Z-bound state; (2) the extensive mutations in helices 1 and 2 destabilize these without affecting the intrinsic stability of helix 3; (3) stabilization and reduced aggregation can be achieved by replacing mutated residues in Z(SPA-1) with the corresponding wild-type Z residues . This stabilization is better correlated to changes in helix propensity than to an expected increase in polar versus nonpolar surface area of the fully folded state.

J Mol Biol, 2004 Jul 23, 340(5), 1131 - 42
Structural characterization of unfolded states of apomyoglobin using residual dipolar couplings; Mohana-Borges R et al.; The conformational propensities of unfolded states of apomyoglobin have been investigated by measurement of residual dipolar couplings between (15)N and (1)H in backbone amide groups . Weak alignment of apomyoglobin in acid and urea-unfolded states was induced with both stretched and compressed polyacrylamide gels . In 8 M urea solution at pH 2.3, conditions under which apomyoglobin contains no detectable secondary or tertiary structure, significant residual dipolar couplings of uniform sign were observed for all residues . At pH 2.3 in the absence of urea, a change in the magnitude and/or sign of the residual dipolar couplings occurs in local regions of the polypeptide where there is a high propensity for helical secondary structure . These results are interpreted on the basis of the statistical properties of the unfolded polypeptide chain, viewed as a polymer of statistical segments . For a folded protein, the magnitude and sign of the residual dipolar couplings depend on the orientation of each bond vector relative to the alignment tensor of the entire molecule, which reorients as a single entity . For unfolded proteins, there is no global alignment tensor; instead, residual dipolar couplings are attributed to alignment of the statistical segments or of transient elements of secondary structure . For apomyoglobin in 8 M urea, the backbone is highly extended, with phi and psi dihedral angles favoring the beta or P(II) regions . Each statistical segment has a highly anisotropic shape, with the N-H bond vectors approximately perpendicular to the long axis, and becomes weakly aligned in the anisotropic environment of the strained acrylamide gels . Local regions of enhanced flexibility or chain compaction are characterized by a decrease in the magnitude of the residual dipolar couplings . The formation of a small population of helical structure in the acid-denatured state of apomyoglobin leads to a change in sign of the residual dipolar couplings in local regions of the polypeptide; the population of helix estimated from the residual dipolar couplings is in excellent agreement with that determined from chemical shifts . The alignment model described here for apomyoglobin can also explain the pattern of residual dipolar couplings reported previously for denatured states of staphylococcal nuclease and other proteins . In conjunction with other NMR experiments, residual dipolar couplings can provide valuable insights into the dynamic conformational propensities of unfolded and partly folded states of proteins and thereby help to chart the upper reaches of the folding landscape.

Cell Microbiol, 2004 Aug, 6(8), 753 - 9
Regulated expression of pathogen-associated molecular pattern molecules in Staphylococcus epidermidis: quorum-sensing determines pro-inflammatory capacity and production of phenol-soluble modulins; Vuong C et al.; Phenol-soluble modulin (PSM) is a peptide complex produced by the nosocomial pathogen Staphylococcus epidermidis that has a strong capacity to activate the human innate immune response . We developed a novel method based on liquid chromatography-mass spectrometry (LC-MS) to quantify the production of the individual PSM components . Each PSM peptide was abundant in most of the 76 S epidermidis strains tested . Importantly, none of the PSM components were secreted by an agr mutant strain, indicating that PSM synthesis is regulated strictly by the agr quorum-sensing system . Furthermore, the agr mutant strain failed to elicit production of TNFalpha by human myeloid cells and induced significantly less neutrophil chemotaxis compared with the wild-type strain . Thus, quorum-sensing in S . epidermidis dramatically influenced activation of human host defence . We propose that an agr quorum-sensing mechanism facilitates growth and survival in infected hosts by adapting production of the pro-inflammatory PSMs to the stage of infection.

Am J Ophthalmol, 2004 Jul, 138(1), 137 - 8
Incidence of acute endophthalmitis after triamcinolone-assisted pars plana vitrectomy; Sakamoto T et al.; PURPOSE: To report the incidence of acute endophthalmitis after triamcinolone acetonide-assisted pars plana vitrectomy (PPV) . DESIGN: A retrospective multicenter interventional case series collected over 23 months . METHODS: We retrospectively reviewed the charts of all patients who underwent triamcinolone-assisted PPV at seven academic clinical centers or eye hospitals from January 2002 to November 2003 . RESULTS: Of a total of 1,886 cases, only 1 case showed acute endophthalmitis due to Staphylococcus epidermidis (0.053%) . No other cases showed any signs of postoperative endophthalmitis . CONCLUSION: Intraoperative use of triamcinolone during PPV is not a high risk factor for acute endophthalmitis.

Pediatrics, 2004 Jul, 114(1), e9 - 15
A controlled study of the relationship between Bordetella pertussis infections and sudden unexpected deaths among German infants; Heininger U et al.; OBJECTIVE: This was a prospective, controlled, multicenter study to investigate the relationship between Bordetella pertussis infections and sudden unexpected deaths among German infants . DESIGN: Between 1995 and 1997, all infants who died at 7 to 365 days of age and for whom autopsies were performed in 1 of 8 participating institutes of legal medicine were enrolled . During a standardized autopsy, nasopharyngeal specimens (NPSs) and tracheal specimens were obtained for polymerase chain reaction (PCR) assays to detect B pertussis . The oligonucleotide primers PTp1 and PTp2, which specifically amplify a 191-base pair DNA fragment of the pertussis toxin operon of B pertussis, were used . Two control subjects (matched according to residence, age, gender, and nationality) were enrolled for each case subject, via a network of pediatricians in private practice, and NPSs were obtained from those infants . Parents of case subjects and control subjects were asked to provide specific information on respiratory illnesses of the child, contact with a known case of pertussis, or close contact with a person with a cough illness during the 4 weeks before death or enrollment, as well as the child's pertussis immunization status . The pathologists performing the autopsies were unaware of the PCR results . RESULTS: Enrolled were 254 infants (66% male) with sudden unexpected deaths and 441 matched control subjects . Autopsies according to protocol were performed for 234 of the case subjects (92%); a diagnosis of sudden infant death syndrome (SIDS) was made for 76% . For the remaining subjects, causes of death were respiratory or other infections (14%), congenital anomalies or organ failures (4%), aspiration (2%), or accidents or traumatic events (4%) . PCR results were positive for B pertussis for 12 case subjects (5.1%) (all with SIDS or respiratory infections) and 5.3% of control subjects . Of the 12 case subjects with positive PCR results, 10 (83%) were male . Questionnaires had been returned by the parents of 5 of the 12 infants . Three had experienced a respiratory illness (all with cough), beginning 7, 14, and 19 days before death . None had a known contact with a case of pertussis . Four of 15 control infants (27%) with positive PCR findings for B pertussis had a cough illness, indicating possible pertussis, and 2 of those 4 developed typical symptoms (whooping) . Background information was received from 116 parents (46%) of case subjects and from parents of all control subjects . Upper respiratory tract infections within 4 weeks before death were reported for 53% of case subjects and 38% of control subjects . Also, fewer case subjects (33%) than control subjects (68%) had received age-adequate numbers of pertussis vaccine doses . CONCLUSIONS: The concept of infection as a factor in SIDS is supported by a number of observations, including the seasonal distribution of the occurrence of SIDS; the high incidence of concurrent upper respiratory tract infections among infants dying as a result of SIDS; the peak age at 3 to 4 months; nicotine use in a child's household, which predisposes children to respiratory infections such as otitis media; and the protective role of breastfeeding . A prominent role might be suspected for B pertussis, for several reasons . 1) B pertussis infections in infancy are frequently associated with apneic spells, which are occasionally life-threatening and, if leading to death, might be reported as SIDS . 2) Epidemiologic evidence from the United Kingdom, Sweden, and Norway indicates that SIDS is associated with B pertussis infection . 3) In a previously published study, we detected B pertussis DNA in the nasopharynx of 9 of 51 consecutive infants (18%) with sudden unexpected deaths . This is the first prospective, controlled study to investigate the possible etiologic role of B pertussis in SIDS . Clinically unrecognized B pertussis infections were relatively frequent (5.3%) among control infants during the course of our study . The rate of infection was similar or perhaps greater for control subjects, compared with case subjects (1.7%), when only NPS results were compared . This may seem surprising but is supported by other studies, in which asymptomatic infections or mild respiratory illnesses were observed among infants exposed to B pertussis . Careful autopsies, including histologic evaluations of organ specimens and use of PCR to detect B pertussis in NPSs and tracheal specimens, represented a strength of this study . Our general findings were as expected . The majority of cases were classified as SIDS . The second largest group included infants for whom respiratory infections were found . The findings of various other diagnoses, which in several instances would have been undiscovered otherwise, emphasize the need for autopsies after unexpected infant deaths . What is the significance of the identified B pertussis infections in 12 cases? Several pieces of evidence support the plausibility of a cause-and-effect relationship . Eight of the 12 case subjects died before 6 months of age, the typical age for death attributable to pertussis . In autopsies, 9 of the subjects were found to have signs of respiratory infections; for 2 infants, the autopsies suggested that death was attributable to a respiratory infection . One additional infant (data not shown) had brain edema (which could have been attributable to hypoxemia during pertussis) . Lower rates of completed primary series or age-adequate numbers of pertussis vaccine doses among case subjects than among control subjects may indicate that immunization against pertussis protects children from death attributable to unrecognized B pertussis infection . Moreover, a recent study indicated that immunization with diphtheria-tetanus-pertussis vaccine induces antibodies that cross-react with pyrogenic staphylococcal toxins, which have been implicated in several cases of SIDS . Other microorganisms may be involved in the sudden death of infants, as suggested in this study by the higher rate of a history of concurrent upper respiratory tract infections among case subjects, compared with control subjects . Similarly, in a Scandinavian study, 48% of 244 SIDS case subjects, compared with 31% of 869 control subjects, exhibited symptoms of upper airway infection during the last week before death or interview, respectively . Because SIDS is a diagnosis of exclusion, every attempt should be made to identify a cause of death during autopsy . This should include the search for pathogenic microorganisms in the respiratory tract with the use of PCR and other sensitive tests . In conclusion, B pertussis infection was found for 12 of 234 infants (5.1%) with unexpected deaths, and the infections might have contributed to the deaths.

Vet Surg, 2004 Jul-Aug, 33(4), 404 - 11
In vitro efficacy of a polyhexamethylene biguanide-impregnated gauze dressing against bacteria found in veterinary patients; Lee WR et al.; OBJECTIVE: To evaluate the in vitro efficacy of polyhexamethylene biguanide (PHMB)-impregnated gauze dressing in limiting the growth of bacteria both within and underneath the dressing . STUDY DESIGN: In vitro study . METHODS: Squares of PHMB-impregnated and control gauze were placed on agar plates inoculated with 1 of 11 bacterial species, including 8 multi-resistant organisms . Growth under the gauze was assessed qualitatively after 24-hour incubation . Repeated use of sponges was used to evaluate residual inhibitory activity against Micrococcus lutea and Staphylococcus schleiferi ss . schleiferi . In a second procedure, PHMB-impregnated and control gauze squares were placed in sterile plastic wells and inoculated with 1 of 5 bacterial species, including Pseudomonas spp . and Klebsiella spp . Inhibition of bacterial growth within and underneath the dressing after 24-hour incubation was evaluated by quantifying the numbers of bacteria on the well floor and within each square . RESULTS: PHMB-impregnated gauze provided greater inhibition of growth of 4/4 Gram-positive species and 2/6 Gram-negative species on inoculated plates compared with control gauze . Residual inhibitory activity of PHMB-impregnated gauze was significantly greater against M . lutea on all days and against S . schleiferi ss . schleiferi on days 1 and 4 compared with control . No bacteria were recovered from inoculated PHMB-impregnated gauze squares placed in sterile wells or from the well floor underneath . More than 9 x 10(5) colony-forming units (CFU) were recovered from inoculated control samples placed in sterile wells and more than 8.4 x 10(4) CFU were recovered from control well floors . CONCLUSION: PHMB-impregnated gauze dressing, when placed on inoculated agar plates, reduces growth of underlying bacteria, particularly Gram-positive species . Wet-inoculated PHMB-impregnated dressing prevents growth of Gram-positive and Gram-negative bacteria both within and underneath the dressing . CLINICAL RELEVANCE: PHMB-impregnated dressings may be useful for reducing contamination of underlying wounds by bacterial pathogens.

Biophys Chem, 2004 Aug 1, 110(3), 239 - 48
Proteins with simplified hydrophobic cores compared to other packing mutants; Chen J et al.; Efforts to design proteins with greatly reduced sequence diversity have often resulted in proteins with so-called molten globule properties . Substitutions were made at six neighboring sites in the major hydrophobic core of staphylococcal nuclease to create variants with all leucine, all isoleucine or all valine at these sites . The mutant proteins with simplified cores constructed here are quite unstable and have poorly packed cores, attested to by interaction energies . Eight related mutants with greater sequence diversity were also constructed . Comparison to these mutants and 159 other permutations of these 3 aliphatic side chains at these same 6 sites previously constructed shows that the simplified cores are not unusual in their stabilities or interaction energies . Further, crystal structures of the two mutants with the worst packing, as measured by interaction energies, showed no unusual disorder in the core . Therefore, reduction of sequence diversity is not necessarily incompatible with a single stable native structure . Other factors must also contribute to previous protein design failures.

Bioorg Med Chem Lett, 2004 Aug 2, 14(15), 3881 - 3
Synthesis and antibacterial activity of arylpiperazinyl oxazolidinones with diversification of the N-substituents; Jang SY et al.; A series of 4-arylpiperazin-1-yl-3-phenyloxazolidin-2-one derivatives with diversification of the N-substituents such as methylene O-linked heterocycles, thioamide, dithiocarbamate, thiourea, and thiocarbamate were synthesized and evaluated as antibacterial agents . Their in vitro activities (MIC) were evaluated against MRSA and VRE resistant Gram-positive strains such as Staphylococcus and Enterococcus . Most of the compounds were more potent in vitro but less active in vivo than linezolid.

Transfusion, 2004 Jul, 44(7), 1098 - 103
Cryopreservation reduces the concentration of detectable bacteria in contaminated peripheral blood progenitor cell products; Kipp F et al.; BACKGROUND: Microbial contamination of PBPC products PBPCPs may cause severe clinical complications . There-fore, we investigated the influence of cryopreservation on the sensitivity to detect bacterial contaminations in PBPCPs . STUDY DESIGN AND METHODS: Expired PBPCPs (n = 29) were thawed, and defined concentrations of Staphylococcus epidermidis or Escherichia coli were inoculated into each bag . After 60 minutes of intermixing, a representative aliquot was drawn and cultured on Mueller-Hinton agar for 24 hours . Then, the products were cryopreserved for 24 hours, and the procedure was repeated as mentioned above . The total numbers of CFUs were counted before and after cryopreservation . RESULTS: A mean concentration of 2529 CFUs per mL of S . epidermidis was determined before cryopreservation versus 2182 CFUs per mL after cryopreservation, demonstrating a decrease of detectable colonies (p < 0.05) . For E . coli, the mean numbers were 424 CFUs per mL before cryopreservation and 343 CFUs per mL after cryopreservation, also showing a decrease (p < 0.05) . CONCLUSION: The cryopreservation reduces the concentration of detectable bacteria in contaminated PBPCPs . Especially in sterility testing of PBPCPs with low bacterial contamination, this phenomenon could lead to false-negative results with severe clinical consequences.

Vasa, 2004 May, 33(2), 83 - 5
Femoral artery infection associated with a percutaneous arterial suture device; Tiesenhausen K et al.; This is a case report of a femoral artery infection with fatal outcome after using a percutaneous suture mediated closure device: A 77-year old patient underwent diagnostic angiography of his thoracic and abdominal aortic aneurysm, the puncture site was closed with the Perclose system . He developed a staphylococcal femoral artery infection with groin abscess, requiring surgical intervention with debridement and removal of the Perclose suture . After stent graft exclusion of the thoracic and abdominal aortic aneurysm a staphylococcal sepsis occurred and the patient died of aneurysm rupture months later despite long term antibiotic therapy . Since the use of the Perclose device carries an increased risk of femoral artery infection with septic endarteritis and bacteremia, it should not be used in routine diagnostic angiography.

Graefes Arch Clin Exp Ophthalmol, 2004 Aug, 242(8), 679 - 81 Epub 2004 Jun 29.
Weak symptoms of bacterial endophthalmitis after a triamcinolone acetonide-assisted pars plana vitrectomy; Yamashita T et al.; PURPOSE: To report a case of endophthalmitis after triamcinolone acetonide (TA)-assisted par plane vitrectomy (PPV) . METHODS: A 60-year-old Japanese man developed endophthalmitis after TA-assisted PPV for diabetic macular edema . Preoperative visual acuity was 20/200 . Four days after surgery, endophthalmitis associated with anterior chamber hypopyon was noticed; the patient's vision had deteriorated to hand motion . In spite of severe cell infiltration, the ciliary injection and ocular pain were not significant . RESULTS: The additional PPV with irrigation of cefazolin (40 microg/ml) and gentamicin (8 microg/ml) was performed . Endophthalmitis resolved soon after this treatment . Staphylococcus epidermidis was detected in the intravitreous samples . The patient's visual acuity improved to 20/100 . CONCLUSION: Endophthalmitis may be a complication of TA-assisted PPV with unique signs and symptoms.

Ann Thorac Surg, 2004 Jul, 78(1), 117 - 20; discussion 120-1
Repair of aortic false aneurysm using deep hypothermia and circulatory arrest; Dumont E et al.; BACKGROUND: Aortic false aneurysms are a rare complication of surgery of the aorta that can occur several months to years after the initial operation . We reviewed our results with false aneurysm repair using deep hypothermia and circulatory arrest . METHODS: Eleven patients were reoperated on for false aneurysm of either the ascending or descending thoracic aorta . Femorofemoral cardiopulmonary bypass with full-dose aprotinin and a heparinized system was used in all patients . Hypothermic circulatory arrest at an average of 20 degrees C was instituted in all patients for repair . Six patients had a patch repair with either polyethylene terephthalate fiber (Dacron) or bovine pericardium, 4 had tube replacement of the aorta, and 1 had primary repair of the defect . RESULTS: Three patients had false aneurysm formation at a site of coarctation repair in the descending aorta, and the 8 others had false aneurysms in the ascending aorta at the site of a previous aortotomy . Six patients had proven infection as the cause; the causative agent was Staphylococcus species in all cases . Mean cardiopulmonary bypass time was 178 +/- 51 minutes, and circulatory arrest time averaged 39 +/- 18 minutes . Operative mortality was 18% (2 of 11); the cause of death was cardiogenic shock in both patients . The mean time to extubation in survivors was 5 days, and the average time to discharge was 16 days . CONCLUSIONS: Although mediastinal infection is a common cause, aortic false aneurysms can be safely approached using femorofemoral cardiopulmonary bypass, hypothermic circulatory arrest, and patch repair with acceptable operative mortality and long-term survival.

Cornea, 2004 Jul, 23(5), 472 - 8
Microbial keratitis following lamellar keratoplasty; Sharma N et al.; OBJECTIVE: To determine the predisposing factors, etiologic agents, and clinical and visual outcomes in infectious keratitis following lamellar keratoplasty (LK) . METHODS: One hundred thirty-five eyes (135 patients) that had undergone LK were retrospectively analyzed for the occurrence of infectious keratitis following LK . The parameters evaluated were predisposing factors, seasonal variation, indications and type of LK, time interval between LK and infection, site and depth of infection, etiologic organisms, type of treatment, outcome in terms of graft status, secondary surgery, visual acuity, and the donor tissue profile . RESULTS: The incidence of infectious keratitis following LK was 11.11% . The most significant predisposing factor was persistent epithelial defect (3 eyes) and suture abscesses (3 eyes) . Most cases occurred between May and August (9/15) . Twelve cases developed infection within 2 weeks of surgery (80%) . Seven cases (7/15) occurred with onlay grafts, 6 with inlay grafts, and 2 with large-diameter LK . Cultures of corneal scrapings were positive in 11 eyes (73.3%), and the most common isolated organism was coagulase-negative Staphylococcus (CNS) . Only 2 eyes responded to medical therapy, and graft sloughing occurred in 9 cases . Six eyes underwent penetrating keratoplasty to either salvage the integrity of globe or for visual rehabilitation of cases where infection resulted in corneal opacity . CONCLUSIONS: Infections after LK may not be amenable to antimicrobial therapy and may necessitate the removal of the graft or a therapeutic penetrating keratoplasty.

J Med Assoc Thai, 2004 Apr, 87(4), 438 - 41
Nocardia pneumonia with empyema thoracis in a healthy neonate: a case report; Tantracheewathorn T et al.; A 40-day-old male infant presented with fever and non-productive cough for 3 weeks, tachypnea and dyspnea 5 days before admission . The chest radiograph and computed tomographic (CT) scan revealed right lung consolidation with pleural effusion . Pleural tapping showed frank pus that grew Nocardia asteroides . The CT scan of the brain and abdomen were normal . The patient was treated with 15 mg/kg/day of trimethoprim and 75 mg/kg/day of sulfamethoxazole intravenously for 3 weeks and then orally for 6 months with complete recovery . T cell quantitative studies were normal and anti-HIV was non-reactive . A flow cytometric assay with Staphylococcal aureus and phorbol myristate acetate (PMA) as the stimulants showed normal oxidative burst which represented normal intracellular killing . One year after completion of treatment, he was healthy and the chest radiograph was normal.

Biosci Biotechnol Biochem, 2004 Jun, 68(6), 1293 - 8
Fluorometric study of the acid-induced denaturation of Staphylococcal nuclease and its mutant forms; Tanaka A; The acid-induced denaturation of wild-type Staphylococcal nuclease (WT) and its eight mutant forms, L25A, V66L, G79S, A90S, G88V, H124L, V66L/G88V, and V66L/G79S/G88V, was investigated using Trp140 fluorescence as a probe at 30 degrees C . The values of pH(1/2), at which the denaturation is half completed, and n, the apparent number of protons which trigger the denaturation and are taken up by the proteins upon denaturation at pH(1/2), were evaluated from the pH dependence of the fluorescence intensity . The values of pH(1/2) and n for WT were 3.8 and 1.8 respectively . The amino acid replacements changed the pH(1/2) values to a range between 3.0 (H124L) and 4.4 (G79S) and also changed the n values to a range between 1.0 (A90S) and 3.0 (G88V) . There was a negative correlation between the values of pH(1/2) and n . It was suggested that the amino acid replacements may change the energy levels of the native state and/or the denatured state mainly in the neutral (stable) pH region, not in the acidic (unstable) region, resulting in the correlative changes in pH(1/2) and n.

Antimicrob Agents Chemother, 2004 Jul, 48(7), 2708 - 11
Purification and characterization of recombinant Staphylococcus haemolyticus DNA gyrase and topoisomerase IV expressed in Escherichia coli; Bronstein JC et al.; The subunits of DNA gyrase and topoisomerase IV from Staphylococcus haemolyticus were expressed in Escherichia coli, purified to homogeneity, and used to reconstitute active enzymes that were sensitive to known topoisomerase inhibitors . This represents the first description of a method for isolating type II topoisomerases of a coagulase-negative staphylococcal species.

J Biomol NMR, 2004 Jul, 29(3), 377 - 85
Correlation times and adiabatic barriers for methyl rotation in SNase; Chatfield DC et al.; The relation of rotational correlation times to adiabatic rotational barriers for alanine methyl groups in staphylococcal nuclease (SNase) is investigated . The hypothesis that methyl rotational barriers may be useful probes of local packing in proteins is supported by an analysis of ten X-ray crystal structures of SNase mutants . The barrier heights are consistent across a set of ten structures of a native SNase and mutants containing single-point mutations or single or double insertions, most in a ternary SNase complex . The barriers for different methyls have a range of 7.5 kcal/mol, which at 300 K would correspond to a five-order-of-magnitude range in correlation time . It is demonstrated that adiabatic rotational barriers can fluctuate significantly during an MD simulation of hydrated SNase, but that a Boltzmann weighted average is predictive of rotational correlation times determined from correlation functions . Even if a given methyl is on average quite sterically hindered, infrequently sampled low-barrier conformations may dominate the Boltzmann distribution . This result is consistent with the observed uniformity of NMR correlation times for (13)C-labeled methyls . The methyl barriers in simulation fluctuate on multiple time scales, which can make the precise relationship between methyl rotational correlation time and methyl rotation barriers complicated . The implications of these issues for the interpretation of correlation times determined from NMR and simulation are discussed.

J Immunol, 2004 Jul 1, 173(1), 297 - 306
Deficiency of CD11b or CD11d results in reduced staphylococcal enterotoxin-induced T cell response and T cell phenotypic changes; Wu H et al.; The beta(2) integrin CD11a is involved in T cell-APC interactions, but the roles of CD11b, CD11c, and CD11d in such interactions have not been examined . To evaluate the roles of each CD11/CD18 integrin in T cell-APC interactions, we tested the ability of splenocytes of CD11-knockout (KO) mice to respond to staphylococcal enterotoxins (SEs), a commonly used superantigen . The defect in T cell proliferation with SEA was more severe in splenocytes from mice deficient in CD18, CD11b, or CD11d than in CD11a-deficient splenocytes, with a normal response in CD11c-deficient splenocytes . Mixing experiments showed that the defect of both CD11b-KO and CD11d-KO splenocytes was, unexpectedly, in T cells rather than in APC . Cytometric analysis failed to detect CD11b or CD11d on resting or activated T cells or on thymocytes of wild-type adult mice, nor did Abs directed to these integrins block responses in culture, suggesting that T cells educated in CD11b-KO or CD11d-KO mice were phenotypically altered . Consistent with this hypothesis, T cells from CD11b-KO and CD11d-KO splenocytes exhibited reduced intensity of CD3 and CD28 expression and decreased ratios of CD4/CD8 cells, and CD4(+) T cells were reduced among CD11b-KO and CD11d-KO thymocytes . CD11b and CD11d were coexpressed on a subset of early wild-type fetal thymocytes . We postulate that transient thymocyte expression of both CD11b and CD11d is nonredundantly required for normal thymocyte and T cell development, leading to phenotypic changes in T cells that result in the reduced response to SE stimulation.

Protein Eng Des Sel, 2004 May, 17(5), 455 - 62 Epub 2004 Jun 18.
Selection and characterization of HER2/neu-binding affibody ligands; Wikman M et al.; Affibody (affibody) ligands that are specific for the extracellular domain of human epidermal growth factor receptor 2 (HER2/neu) have been selected by phage display technology from a combinatorial protein library based on the 58 amino acid residue staphylococcal protein A-derived Z domain . The predominant variants from the phage selection were produced in Escherichia coli, purified by affinity chromatography, and characterized by biosensor analyses . Two affibody variants were shown to selectively bind to the extracellular domain of HER2/neu (HER2-ECD), but not to control proteins . One of the variants, denoted His6-ZHER2/neu:4, was demonstrated to bind with nanomolar affinity (approximately 50 nM) to the HER2-ECD molecule at a different site than the monoclonal antibody trastuzumab . Furthermore, radiolabeled His6-ZHER2/neu:4 affibody showed specific binding to native HER2/neu, overexpressed on the SKBR-3 tumor cell line . Such affibody ligands might be considered in tumor targeting applications for radionuclide diagnostics and therapy of adenocarcinomas such as breast and ovarian cancers.

Poult Sci, 2004 Jun, 83(6), 1047 - 50
Effects of applying Safe2O poultry wash to broiler wings on shelf life, Listeria monocytogenes, Pseudomonads, Staphylococcus species, and psychrotrophic bacteria levels after three, seven, and ten days of storage; Dickens JA et al.; Bacterial contamination of raw processed poultry continues to be of concern to consumers as well as regulatory and health officials . For many years wings were considered a low-value product; therefore, shelf life of wings was not a major concern . Due to changes in consumer attitudes and increases in the fast-food market, wings are now a valuable commodity . Because wings have a shorter shelf life than most other raw poultry products, acceptable intervention to decrease the population of associated spoilage organisms and human enteropathogens are needed . Safe2O Poultry Wash was evaluated as a postchill treatment to reduce microbial contamination and increase shelf life . Ninety-six carcasses were obtained from a local processor prior to final wash . On arrival at the research facility all carcasses were inoculated with 1 mL of a culture with 10(3) cfu/mL Listeria monocytogenes . After a 30-min attachment time, carcasses were subjected to a 4-s in-out final wash, hung for 3 min, and chilled in ice-water for 45 min . After the chilling, wings were removed by hand with a knife, pooled together, and subjected to a hand spray (4 mL/wing) with deionized water or Safe2O Poultry Wash . Two wings were then placed in each of 96 ziplock type storage bags, and wings were held at 5 +/- 1 degrees C for 3, 7, 10, and 14 d . On the day of sample, weep was decanted, and 100 mL of Butterfield's phosphate buffer was added to each bag . Three sets of wings were shaken by hand for 1 min, and total aerobes, Pseudomonads, Staphylococcus sp., psychrotrophic bacteria, and L . monocytogenes in the rinsates were enumerated . By using 7 log10 recovery of total aerobes from rinsates as a spoilage baseline, all wings were spoiled by d 10, but the wings treated with water were approaching spoilage counts on d 7, (log10 6.8), whereas only log10 5.5 bacteria were recovered from the wings sprayed with Safe2O Poultry Wash . Fewer Pseudomonads, Staphylcoccus sp., L . monocytogenes, and psychrotrophic bacteria were recovered from wings treated with Safe2O Poultry Wash and stored for 10 d . Log10 counts for the organisms were Pseudomonas sp., 8.2 and 6.9; Staphylcoccus sp., 5.5 and 4.9; L . monocytogenes, 5.2 and 4.6; and psychrotrophs, 8.2 and 6.9 for the water and Safe2O Poultry Wash treatments, respectively . Use of the Safe2O Poultry Wash as a postchill treatment on wings could increase the shelf life of wings by up to 3 d.

Transpl Immunol, 2004 Jun-Jul, 13(1), 55 - 61
Ex vivo lymphocyte proliferative function is severely inhibited in renal transplant patients on mycophenolate mofetil treatment; Hutchinson P et al.; Mycophenolate mofetil (MMF) is a recently introduced immunosuppressive drug . Its active form is mycophenolic acid (MPA) . MPA specifically inhibits de novo purine synthesis, which is vital for T and B lymphocyte proliferation . We measured lymphocyte subset numbers and mitogen induced proliferation in kidney transplant recipients on different combinations of MMF, cyclosporin A (C), azathioprine (A) and prednisolone (P) (C+A n=70; C+A+P n=15; C+MMF n=45; C+MMF+P n=37) and normals (n=73) . Patients on MMF had severely reduced phytohaemagglutinin A (PHA) induced proliferation compared to normals (Nml 2766+/-926 CPM/1000 lymphocytes {mean+/-S.D.}; C+MMF 282+/-406; C+MMF+P 195+/-496); non-MMF patients did not differ from normal . Similar inhibition of Poke Weed Mitogen and Staphylococcal enterotoxin B induced proliferation was observed . Cell cycle studies established that MMF patients had a significantly higher proportion of lymphocytes in the G0/G1 phase following PHA stimulation than the non-MMF patients . All transplant groups had significantly lower B cell numbers than the normal controls but no differences in CD4 and CD8 T cell numbers . All but the C+MMF group had significantly lower CD16+NK cell numbers than normal, while only the non-MMF groups had significantly lower CD56+NK cell numbers . The proliferation assay used was an ex vivo diluted whole blood technique . Removal of residual MPA by washing the plasma out prior to mitogen stimulation led to a significant increase in proliferation in six out of seven cases . In summary we have found that MMF treatment has a strikingly inhibitory effect on patient ex vivo lymphocyte mitogenic function.

Transplantation, 2004 Jun 15, 77(11), 1699 - 706
P-glycoprotein activity is decreased in CD4+ but not CD8+ lung allograft-infiltrating T cells during acute cellular rejection; Donnenberg VS et al.; BACKGROUND: Modulation of P-glycoprotein (P-gp) activity in graft-infiltrating T cells may alter their susceptibility to immunosuppression . METHODS: P-gp activity was measured by rhodamine efflux in T-cell subsets from bronchoalveolar lavage (BAL) of five healthy volunteers and 27 lung allograft recipients . The effect of T-cell activation on P-gp activity was modeled by stimulation of peripheral blood mononuclear cells with staphylococcal enterotoxin B . RESULTS: Most BAL T cells expressed memory-effector markers . Patients had a lower proportion of CD4 T cells (P = 0.005), whereas control subjects had CD4-to-CD8 ratios similar to peripheral blood . In controls, basal P-gp activity was greatly increased in both CD4 (35% P-gp active) and CD8 (63%) lung T cells compared with peripheral T cells . Basal P-gp activity was elevated in patient BAL T cells but was lower than control BAL activity (CD4, P = 0.07; CD8, P = 0.03) . Lung T cells from transplant patients had modest (CD4) or marked (CD8) increases in substrate-induced P-gp activity compared with normal lung, indicating that P-gp was not irreversibly inhibited . Patients with acute cellular rejection (ACR) had reduced P-gp activity in CD4, but not CD8, BAL T cells compared with patients without ACR (P = 0.004) . To determine the relationship between T-cell activation on P-gp modulation, P-gp activity was measured in staphylococcal enterotoxin B-stimulated peripheral blood mononuclear cells . P-gp activity was abrogated in CD71 cycling cells but remained high in a persistent but minor population of resting naive T cells . CONCLUSIONS: Lung T cells have increased in vivo P-gp activity and therefore may eliminate substrate drugs, resulting in local resistance to immunosuppressive therapy . However, P-gp function is reduced during T-cell activation, providing a window of susceptibility to treatment during ACR.

J Pept Res, 2004 Jul, 64(1), 33 - 41
Purification and characterization of a new peptide with analgesic effect from the scorpion Buthus martensi Karch; Cao ZY et al.; Anew peptide, designated as Buthus martensi Karch (BmK) AngM1, with an isoelectric point (pI) of 5.8 was purified and characterized from the venom of Buthus martensi Karch . The molecular mass was calculated as 7040.5 Da from multiple-charged ions by elelctrospray ionization mass spectroscopy (ESI/MS) . The complete amino acid sequence of BmK AngM1 of 64 amino acid residues was determined by automatic sequencing of N-terminal part of the native peptide and the fragments of reduced and S-carboxymethylated (RCM)-peptide degraded by Staphylococcus aureaus V(8) protease and TPCK(N-p-Tosyl-L-phenylalanine chloromethyl ketone)-treated trypsin . Bioactivity tested using mouse-twisting model showed an evident analgesic effect with 63.0% (P < 0.001) inhibition efficiency at the dose of 0.8 mg/kg, but the LD(50) was larger than 50 mg/kg . Electrophysiological studies showed that BmK AngM1 at the concentration of 1 microm obviously inhibit voltage-dependent Na(+) current (I(Na)) and voltage-dependent delayed rectifier K(+) current (I(K)) but had no effects on transient K(+) current.

Zhonghua Er Bi Yan Hou Ke Za Zhi, 2004 Feb, 39(2), 93 - 6
{Expression of substance P receptor positive cells in the Corti's organ with acute middle ear infection in guinea pigs}; Qiu JH et al.; OBJECTIVE: To investigate the expression of substance P receptor(SPR), SPR positive cells in the Corti's organ with acute middle ear infection in guinea pigs using the polyclonal antibody of SPR . METHODS: Twelve healthy guinea pigs were employed in the experiment . After general anesthesia, by injecting 1 x 10(8)/L staphylococcosis aureus into the middle cavity of right ear with the left ear serving as control, the acute middle ear infection model was established . Then three days later, immunohistochemical staining of SPR was performed in the Cochlear base membrane preparation . RESULTS: Microscopic examination of whole cochlear preparation revealed a number of SPR positive cells expression in the cochlear base membrane, these labeled cells usually did not exist in normal cochlear tissue . Obvious difference in morphology and distribution could be identified with inner hair cells, outer hair cells vascular endothelial cell and spiral ganglion neurons . Labeled SP receptor positive cells were similar to "neurons", scattering distributed in the free margin of cochlear base membrane, with larger size and multiple projections which was 6-12 times than the erythrocyte . There were the vesicle and granular substances in the cytoplasm of the labeled cells . CONCLUSIONS: Acute middle ear infection could induce the expression of SP receptor positive nonspecific cells in the Corti's organ of guinea pigs . These cells did not exist in the normal base membrane and might participate in initiating or inducing the immune response of inner ear.

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi, 2004 May, 20(3), 360 - 2
{Preparation and the anti-tumor activity of mutant Staphylococcal enterotoxin B}; Deng XH et al.; AIM:To prepare mutant Staphylococcal enterotoxin B(SEB) and observe its anti-tumor activity . METHODS: The expressed mutant SEB-K172E in inclusion body was denatured and renatured, and then isolated and purified . The anti-tumor activity of the mutant SEB-K172E was compared with wild-type SEB . RESULTS: Renaturation and purification method of the mutant SEB-K172E was developed . The anti-tumor activity of acquired mutant protein was ten times higher than that of wild-type SEB . CONCLUSION: The anti-tumor activity of the mutant SEB-K172E is much higher than that of wild-type SEB, suggesting that the mutant SEB-K172E may become a potential anti-tumor drug.

J Intensive Care Med, 2003 Jan-Feb, 18(1), 47 - 51
Pericardial infusion of tissue plasminogen activator in fibropurulent pericarditis; Reznikoff CP et al.; A 61-year-old man developed a loculated fibropurulent pericarditis, a rare complication of bacteremia . This occurred as a complication of a Staphylococcal aureus bacteremia from a head and neck abscess following self-extraction of a tooth . Despite surgical intervention and placement of 2 pericardial drains, a refractory, inadequately drained infected pericardial effusion persisted . Although there is limited experience with thrombolytic therapy to dissolve a fibrin clot in the pericardium, break down loculated adhesions, and facilitate free drainage of infected material, lysis is well described in the management of exudative pleural effusions . After infusion of 30 mg of tissue plasminogen activator in 100 cc normal saline through the pericardial drain of the patient, a large amount of infected serosanginous material subsequently drained during the next 2 days . The patient became afebrile and culture negative, remained hemodynamically stable, and had resolution of his pericarditis and pericardial effusion on electrocardiogram and echocardiogram, respectively.

Atherosclerosis, 2004 Jul, 175(1), 83 - 90
Statin-induced immunomodulatory effects on human T cells in vivo; Fehr T et al.; Statins are widely used for treatment of hypercholesterolemia . Recent experimental studies revealed that these drugs also exert anti-inflammatory effects . The aim of this study was to assess immunomodulatory effects of statins in humans in vivo . Twenty-seven healthy volunteers were analyzed for serum cytokines and acute phase proteins, HLA-DR and CD38 expression on T cells and superantigen-mediated T cell activation ex vivo before and after 14 days of statin treatment . First, simvastatin 40 mg was compared to atorvastatin 20 mg . Second, two different doses of simvastatin (20 and 40 mg) were tested . Atorvastatin treatment led to a significant down-regulation of HLA-DR and the CD38 activation marker on peripheral T cells, whereas simvastatin up-regulated both of these molecules . In contrast, superantigen-mediated T cell activation was inhibited by simvastatin and enhanced by atorvastatin . No significant effect of statin treatment on inflammatory serum markers was detected . Thus, immunomodulatory effects of statins on human T cells are first demonstrated in vivo and are differentially induced by two different statins: atorvastatin led to a major histocompatibility class II (MHC II) antigens down-regulation and may therefore be investigated for treatment of chronic transplant rejection; simvastatin inhibited superantigen-mediated T cell activation, which might explain reduced mortality of simvastatin-treated patients with staphylococcal bacteremia.

Appl Environ Microbiol, 2004 Jun, 70(6), 3263 - 71
Localization and functional analysis of PepI, the immunity peptide of Pep5-producing Staphylococcus epidermidis strain 5; Hoffmann A et al.; Pep5 is a cationic pore-forming lantibiotic produced by Staphylococcus epidermidis strain 5 . The producer strain protects itself from the lethal action of its own bacteriocin through the 69-amino-acid immunity peptide PepI . The N-terminal segment of PepI contains a 20-amino-acid stretch of apolar residues, whereas the C terminus is very hydrophilic, with a net positive charge . We used green fluorescent protein (GFP)-PepI fusions to obtain information on its localization in vivo . PepI was found to occur outside the cytoplasm and to accumulate at the membrane-cell wall interface . The extracellular localization appeared essential for conferring immunity . We analyzed the functional role of the specific segments by constructing various mutant peptides, which were also fused to GFP . When the hydrophobic N-terminal segment of PepI was disrupted by introducing charged amino acids, the export of PepI was blocked and clones expressing such mutant peptides were Pep5 sensitive . When PepI was successively shortened at the C terminus, in contrast, its export properties remained unchanged whereas its ability to confer immunity was gradually reduced . The results show that the N-terminal part is required for the transport of PepI and that the C-terminal part is important for conferring the immunity phenotype . A concept based on target shielding is proposed for the PepI immunity mechanism.

Cutis, 2004 May, 73(5 Suppl), 19 - 20
Staphylococcal resistance and oral cephalosporins for skin and skin structure infections; Joseph WS; The common classification of cephalosporins into "generations," based on their activity toward gram-positive and gram-negative organisms, does not adequately reflect the differences among these drugs, especially for newer, broad-spectrum antibiotics . Thus, clinicians need to understand the complete spectrums of activity for these drugs to treat patients effectively and minimize the development of drug resistance.

J Infect Dis, 2004 Jun 15, 189(12), 2334 - 6 Epub 2004 May 26.
Standard nomenclature for the superantigens expressed by Staphylococcus; Lina G et al.; The International Nomenclature Committee for Staphylococcal Superantigens proposes an international procedure for the designation of newly described superantigens and putative superantigens, a procedure that will be compatible with the new age of genomics.

J Chromatogr B Analyt Technol Biomed Life Sci, 2004 Jul 25, 807(1), 95 - 103
Decontamination of surfaces by lysozyme encapsulated in reverse micelles; Bermudez O et al.; Cells and enzymes can be used to decontaminate soil, water supplies, personal equipment, weapons and hospital equipment that have been exposed to bacteria, toxins or viruses . One of the problems associated with the use of microorganisms and enzymes for decontamination purposes is that the presence of water is not acceptable for some applications such as electronic equipment . One way of circumventing this problem is to allow the enzyme to distribute between a water phase and an organic phase-containing surfactant and then use the encapsulated enzyme in reverse micelles directly into the device to be clean . Reverse micelles were used to deliver the enzyme (lysozyme) to the cell-surface interface . They serve as a way to increase the local concentration of lysozyme and decrease the amount of water delivered . Specifically, we explored the lysis by free lysozyme and lysozyme encapsulated in reverse micelles of Klebsiella pneumoniae and Staphylococcus epidermidis attached to steel, glass, and hydroxyapatite . These two bacteria have been selected because they are known to be pathogenic and because of their differences in cell wall structure . Lysozyme was added to the surfaces in either reverse micelles or as a free solution and was tested under conditions of stirring and no stirring . Stirring was implemented to study the interplay between mass transfer limitations and surface roughness . We have shown that free lysozyme or lysozyme encapsulated in reverse micelles is capable of decontaminating surfaces of different texture . Lysis of the cells is slower when the encapsulated enzyme is used but lysis is more complete.

Am J Infect Control, 2004 Jun, 32(4), 232 - 4
Validation of a multicenter computer-based surveillance system for hospital-acquired bloodstream infections in neonatal intensive care departments; Graham PL 3rd et al.; Automated systems can facilitate surveillance for health care-associated infections . The New York Antimicrobial Resistance Project (NYARP) electronically monitors trends in bloodstream infections from 6 medical centers in New York, NY . To validate NYARP's data, episodes of health care-associated bloodstream infections detected by this system were compared with those obtained by an infection control practitioner performing an unrelated study in 2 participating neonatal intensive care departments . The sensitivity (84%), specificity (99%), and positive (84%) and negative (99%) predictive values of NYARP were excellent when coagulase-negative staphylococcal bloodstream infections were removed.

Curr Allergy Asthma Rep, 2004 Jul, 4(4), 276 - 84
Keratinocytes in atopic dermatitis: inflammatory signals; Esche C et al.; Atopic dermatitis (AD) is a chronic inflammatory skin disorder that usually predates the development of allergic airway disease . In most cases, this is thought to be an allergen-driven disease with prominent roles played by antigen presenting cells and effector Th2 cells . But keratinocytes, by virtue of their location, provide an important window to the environment and are also thought to contribute to the development of AD . In this review, we discuss several biologic attributes of keratinocytes that are relevant for AD: 1) intrinsic defects in barrier function, 2) production of inflammatory mediators that promote or maintain allergic inflammation, 3) keratinocyte apoptosis, 4) effects of staphylococcal toxins on keratinocytes, and 5) potential consequences of the expression of cosignaling molecules (eg, B7 family members) and receptors important for innate immune responses (eg, Toll receptors) . Clearly, these findings have highlighted a more active role played by the epithelium than was previously recognized.

Acta Neurochir (Wien), 2004 Jun, 146(6), 603 - 10; discussion 609-10 Epub 2004 Apr 08.
Effect of hydrophilic coating on microorganism colonization in silicone tubing; Cagavi F et al.; BACKGROUND: Shunt infections are one of the major causes of mortality and morbidity of patients with hydrocephalus . The aim of this research is to compare the bacterial colonization characteristics of a regular silicone elastomer shunt material coated with polyvinylpyrrolidone and dimethylpolysiloxane (silicone) . METHOD: Regular coated shunt materials were compared by in-vivo and in-vitro methods . In the in-vitro experiment, silicone and coated material immersed and not immersed in vancomycin solution was treated with a certain concentration of Staphylococcus epidermidis . In the in-vivo study, silicone and coated material specimens were treated with Staphylococcus epidermidis and they were stereotactically placed in the lateral ventricles of the rats . One week after the inoculation, shunt pieces were removed and the colonies were counted by using a scanning electron microscope . FINDINGS: There was a statistically significant difference of colonization in the in-vitro groups in coated material vs . silicone, coated material vs . vancomycin treated silicone, vancomycin treated coated material vs . silicone, vancomycin treated coated material vs . vancomycin treated silicone . There was no statistically significant difference for colonization in in-vitro groups of coated material and vancomycin treated coated material . With in-vivo experiments we can say that, coated material catheters are superior than the silicone catheters in respect to colonization but after the bacterial colonization has occurred, the amount of colonization did not differ . INTERPRETATION: Coated material catheters are superior to silicone catheters and they prevent bacterial colonization in some respect .

Curr Opin Infect Dis, 2004 Jun, 17(3), 237 - 41
Coagulase-negative staphylococcal disease: emerging therapies for the neonatal and pediatric patient; Weisman LE; PURPOSE OF REVIEW: Due to the high frequency, significant clinical impact, and cost of coagulase-negative staphylococcal infections, a great deal of effort continues in the investigation of the epidemiology, prevention, and treatment of coagulase-negative staphylococcal infection in the neonate and infant . RECENT FINDINGS: Pediatric oncology and burn patients appear to be high-risk groups for coagulase-negative staphylococcal infections . Coagulase-negative staphylococcus appears to be the major pathogen world-wide, and associated with significant morbidity and mortality in neonatal intensive care units, in pediatric intensive care units, and following ophthalmologic surgery . A screened human polyclonal antibody and a humanized chimeric monoclonal antibody are both under investigation (in phase II-III clinical trials) for prevention of coagulase-negative staphylococcal infections in high-risk neonates . Resistance of coagulase-negative staphylococcus to antibiotics appears to be increasing . Should neonates with sepsis and central catheters have their catheters removed immediately or only when the infection is persistent? Arbekacin and linezolid are two new antibiotics that appear to be effective in the treatment of coagulase-negative staphylococcal infections, but their precise role has not yet been identified . SUMMARY: This article summarizes the significant clinical reports about coagulase-negative staphylococcal infections since December 2002.

Alcohol Clin Exp Res, 2004 May, 28(5), 824 - 8
Acute alcohol consumption inhibits accessory cell function of monocytes and dendritic cells; Szabo G et al.; BACKGROUND: Alcohol affects both innate and acquired immune responses . Chronic alcoholics have reduced delayed-type hypersensitivity response and increased susceptibility to infections . In contrast, recent studies suggest that acute, moderate alcohol consumption has protective effects on mortality . Monocytes and dendritic cells (DC) play a central role in coordination of innate and adaptive immune responses and are pivotal in activation of T lymphocytes in an antigen-specific manner . In this study, we investigated the effects of acute, moderate alcohol consumption on antigen presenting cell function of blood monocytes and monocyte-derived myeloid dendritic cells . METHODS: Accessory cell function of human blood monocytes was tested before and after acute alcohol intake (2 ml vodka/kg body weight) by measuring T cell activation with alloantigen (mixed lymphocyte reaction, MLR), superantigen (staphylococcal enterotoxin B) and recall antigen (tetanus toxoid) . Myeloid DCs were generated in vitro from monocytes obtained from these individuals using IL-4 and GM-CSF and their allostimulatory function was tested in an MLR . RESULTS: We found significantly reduced T cell proliferation in the presence of monocytes obtained 2 or 18 hr after alcohol consumption whether alloantigen, superantigen, or recall antigen was the stimuli (p < 0.01) . The reduced T cell proliferation was due to the effects of acute alcohol on monocytes rather than on T cells as we found decreased proliferation only in the presence of alcohol-exposed accessory cells but not when T cells were exposed to alcohol . In addition, monocyte-derived dendritic cells showed significantly reduced allostimulatory capacity after alcohol consumption (p < 0.005) . CONCLUSION: Acute alcohol consumption inhibits accessory cell function of both monocytes and myeloid dendritic cells . Impaired function of these key antigen-presenting cells may contribute to reduced adaptive immune responses and increased susceptibility to infections when acute alcohol intake coincides with exposure to pathogens.

Hybrid Hybridomics, 2004 Apr, 23(2), 133 - 6
Production of monoclonal antibodies against a 19-kD recombinant Plasmodium vivax MSP1 for detection of P . vivax malaria in Turkey; Ak M et al.; Plasmodium vivax malaria, which is transmitted to humans by mosquitoes, is one of the most important parasitic diseases in Turkey . The major protein on the surface of asexual erythrocytic stage merozoites of P . vivax (Pv) is 200 kD and called major merozoite surface protein-1 (PvMSP1) . Polyclonal antibodies against the 19-kD C-terminal fragment of PvMSP1 (PvMSP1(19)) are protective in monkey models of P . vivax and associated with protection in field studies . In this research, monoclonal antibodies were produced against PvMSP1(19) . A total of 214 IgG(1) antibody-releasing hybridomas were obtained and three monoclonal antibodies were produced (PvMSP1(19).1, PvMSP1(19).2, and PvMSP1(19).3) and selected for further study . They have now been purified from ascitic fluid on a Staphylococcus protein A affinity column.These are the first monoclonal antibodies produced against P . vivax in Turkey and the first monoclonal antibodies produced against this recombinant PvMSP1(19) in the world . The monoclonal antibodies will be used to study the epidemiology of P . vivax in patients with malaria in Turkey, and to develop better strategies for early diagnosis and treatment of the disease in our population.

Int J Antimicrob Agents, 2004 Mar, 23(3), 226 - 34
Effects of linezolid on staphylococcal adherence versus time of treatment; Pagano PJ et al.; Staphylococcus epidermidis has emerged as a major nosocomial pathogen that is often associated with infections of indwelling medical devices . Microbial adhesion to implanted foreign materials is a prerequisite for establishing infection . We studied the time-dependent anti-adhesion effects of linezolid and vancomycin on three S . epidermidis clinical isolates . Minimum inhibitory concentration (MIC) values were identical for both agents for all three isolates (2 mg/l) . Bacterial suspensions were added to polystyrene wells and treated with 0.5-4 times the MIC of linezolid or vancomycin at 0, 2, 4 or 6 h post-inoculation . Supra-inhibitory (2 and 4 x MIC) and inhibitory (MIC) concentrations of linezolid demonstrated potent anti-adhesion activity following 2 and 4 h deferred treatments . Even at sub-inhibitory concentrations (0.5 x MIC), suppression of staphylococcal adherence to polystyrene was still evident in most cultures . Linezolid at two and four times the MIC also exerted significant inhibitory effects in cultures that had been treated with a 6-h delay . Supra-inhibitory and inhibitory concentrations of vancomycin administered 2 h post-infection appeared equally effective as linezolid . However, sub-inhibitory concentrations of vancomycin showed minimal or no activity against bacterial adhesion . When vancomycin treatments were delayed by 4 h, only concentrations above the MIC prevented adherence . Linezolid has promising in vitro anti-adhesion activity that merits further studies to determine its role in the management of foreign-body infections.

Antibiot Khimioter, 2004, 49(1), 22 - 5
{Use of clarithromycin in the treatment of atopic dermatitis complicated by staphylococcal infection}; Gintovt EA; Thirty patients (19 boys and 11 girls) with atopic dermatitis complicated by pyodermia were treated with clarithromycin tablets (Fromilid, KRKA, Slovenia) in a dose of 7.5 mg/kg body weight twice a day . The average treatment course was 7 days . The severity of atopic dermatitis was evaluated by the SCARAD Index . Thirty patients of the reference group were treated with intramuscular lincomycin in a dose of 10-20 mg/kg body weight twice a day for 7 days . The difference in the average SCORAD Index before and after the treatment with clarithromycin amounted to 19.95 vs 14.68 (p < 0.005) with the use of lincomycin . The tolerance of clarithromycin was good.

Indian J Pediatr, 2004 May, 71(5), 433 - 5
Toxic shock syndrome; Dass R et al.; The authors present two children who had fever >or=38.9 degree C, diffuse rash, hypotension, deranged renal and hepatic functions, disseminated intravascular coagulation, altered sensorium and inflamed oral mucosa . They responded to fluids, inotropes, antibiotics and intravenous immunoglobulin (2 g/kg) . Desquamation particularly of palms and soles and periungal region was noted 1 to 2 weeks after onset of illness . These features were consistent with the diagnosis of staphylococcal toxic shock syndrome (TSS) . The cases highlight that TSS is very much with us and can mimic a variety of other diseases . Early recognition, and aggressive antimicrobial supportive and IVIG therapy cover can ensure complete recovery.

Eur J Immunol, 2004 Jun, 34(6), 1762 - 70
Activated T killer cells induce apoptosis in lung epithelial cells and the release of pro-inflammatory cytokine TNF-alpha; Shinbori T et al.; Apoptosis is thought to be involved in lung epithelial cell damage in acute respiratory distress syndrome and interstitial pneumonia . Both the role of apoptosis and its underlying molecular mechanisms in human lung tissue remain unclear . To address these issues, we developed an in vitro assay in which a human lung epithelial cell line and a staphylococcal enterotoxin B (SEB)-reactive human CD8(+) CTL line were co-cultured in the presence of SEB . SEB-stimulated CD8(+) CTL induced apoptosis in the lung epithelial cell line primarily through the perforin/granzyme-mediated pathway . In these cells, apoptosis was initially independent of death receptor pathways . We also tested the effect of IFN-gamma on modulation of apoptosis in lung epithelial cells . In IFN-gamma-pretreated lung epithelial cells, CD95 (APO-1/Fas) activation as well as TNF-related apoptosis-inducing ligand (TRAIL) receptor and TNFR activation led to apoptosis . Furthermore, we found that the interaction of SEB-stimulated CD8(+) CTL with lung epithelial cells induced an increase in TNF-alpha secretion . These results suggest an important role for bacterial superantigen-reactive CD8(+) CTL in induction of lung epithelial cell apoptosis and in modulation of inflammatory processes in lung tissue.

FEMS Microbiol Lett, 2004 Jun 1, 235(1), 1 - 8
Staphylococcal methicillin resistance: fine focus on folds and functions; Mallorqui-Fernandez G et al.; Globalisation has entailed a massive increase in trade and human mobility facilitating the rapid spread of infectious agents, including those that are drug resistant . A particularly serious threat to human health is posed by methicillin-resistant staphylococcal strains which have acquired molecular mechanisms to evade the action of beta-lactam antibiotics (BLAs) . Full expression of high-level methicillin resistance involves a complex network of molecules and depends primarily on sufficient expression of a penicillin-binding protein with low sensitivity towards BLAs . Other factors include the fine-tuned regulation of autolytic activity of cell-wall components, as well as an optimal rate of peptidoglycan precursor formation and a highly specific peptidoglycan precursor structure . Three-dimensional structural data are available on several of the pieces involved in the jigsaw puzzle and provide a molecular basis for the understanding of methicillin resistance and for the design of new therapeutic strategies.

Transfusion, 2004 Jun, 44(6), 877 - 85
Photochemical inactivation of selected viruses and bacteria in platelet concentrates using riboflavin and light; Ruane PH et al.; BACKGROUND: A medical device is being developed for the reduction of pathogens in PLT concentrates (PCs) . The device uses broadband UV light and the compound riboflavin (vitamin B(2)) . STUDY DESIGN AND METHODS: Pathogens were added to single-donor PLTs . After treatment, the infectivity of each pathogen was measured using established biologic assays . In vitro PLT performance was evaluated after treatment and after 5 days of storage using a panel of 10 in-vitro cell quality assays . RESULTS: In studies with viral pathogens, the Pathogen Reduction Technology (PRT) system provided average log reduction factors of 4.46 +/- 0.39 for intracellular HIV, 5.93 +/- 0.20 for cells associated HIV, and 5.19 +/- 0.50 for West Nile virus . For the nonenveloped porcine parvovirus, a reduction factor greater than 5.0 log was observed . Staphylococcus epidermidis and Escherichia coli bacteria were also tested with observed reduction factors to the limits of detection of 4.0 log or greater . PLT cell quality was adequately maintained after treatment and during storage . Although P-selectin expression, glucose consumption, and lactate production increased relative to controls, this was not beyond accepted levels . The pH of treated PCs also decreased slightly relative to control PLTs on Days 1 and 5 . CONCLUSION: The data indicate that the device successfully reduced the number of selected pathogens in PCs . Despite the fact that significant differences exist between treated and control in-vitro variables, it is speculated that the clinical effectiveness of both products will not be significantly different, based on comparison to historical data for products in routine clinical use today.

Infect Immun, 2004 Jun, 72(6), 3664 - 7
Biological properties of staphylococcal enterotoxin-like toxin type R; Omoe K et al.; We investigated the biological properties of a novel staphylococcal enterotoxin-like putative toxin, staphylococcal enterotoxin-like toxin type R (SElR) . Major histocompatibility complex class II molecules were required for T-cell stimulation by SElR . SElR stimulated T cells bearing receptors Vbeta 3, 11, 12, 13.2, and 14 . These results suggested that SElR acts as a superantigen.

Pediatr Int, 2004 Jun, 46(3), 337 - 41
Staphylococcal enterotoxin-specific IgE antibodies in atopic dermatitis; Ide F et al.; BACKGROUND: The authors clarified the clinical significance of the measurement of serum concentrations of specific IgE antibodies to staphylococcal enterotoxin (SE) A- and SEB in atopic dermatitis (AD) . METHODS: The serum concentrations of SEA- and SEB-specific IgE antibodies in 140 pediatric patients with AD were measured with an immuno CAP -radioallergosorbent test system (RAST) . To check the cross-reaction of specific IgE antibodies to SEA/SEB and other allergens, the CAP RAST fluorescent enzyme immunoassay inhibition test was performed . RESULTS: Forty-seven patients (33.6%) tested positive for either SEA- or SEB-specific IgE antibodies . School children showed higher positive rates of SEA/SEB-specific IgE antibodies than infants or young children . The patients with severe AD and those with exacerbation of symptoms in summer, had higher positive rates of SEA/SEB-specific IgE antibodies than patients with mild AD or those with exacerbation in winter . In addition, the positive rates of specific IgE antibodies to both dog-dander and cat-dander were higher in patients with positive SEA/SEB-specific IgE antibodies than in patients with negative ones . No cross-reactions occurred among specific IgE antibodies to SEA/SEB and dog/cat dander with one patient's serum, which had positive IgE-specific antibodies against cat/dog dander and SEA/SEB . The positive rate of SEA/SEB-specific IgE antibodies in the patients with dogs and/or cats as pets was 48.4%, which was higher than in those with no pets . CONCLUSIONS: Atopic dermatitis patients who exhibit high positive rates of SEA/SEB-specific IgE antibodies were found to be school children, severe cases, cases with high serum concentrations of total IgE, cases with exacerbation in summer, and cases with dogs and/or cats as pets . The measurement of serum concentrations of specific IgE antibodies to SEA and SEB, thus has some value for evaluating AD patients.

J Med Microbiol, 2004 Jun, 53(Pt 6), 545 - 9
Genetic polymorphism of the accessory gene regulator (agr) locus in Staphylococcus epidermidis and its association with pathogenicity; Li M et al.; Staphylococcus epidermidis has become one of the most important causes of nosocomial infections in recent years . The staphylococcal accessory gene regulator (agr) is the most important locus responsible for the regulation of virulence factors, and it has been shown to be polymorphic . The aim of this study was to investigate the agr locus and its genetic polymorphisms in different Chinese S . epidermidis isolates and the relationship between genetic polymorphisms and pathogenicity . Specific PCR was used to amplify the different agr groups . Results were confirmed by restriction enzyme digestion and sequence analysis . agr mutations were detected and three agr groups of S . epidermidis were determined . Of the isolates, 12 % were pathogenic S . epidermidis and 17 % had naturally occurring agr mutations (P > 0.05) . Pathogenic S . epidermidis isolates comprised 68.2 % agr group I, 19.3 % group II and 12.5 % group III, while isolates from healthy controls comprised 39 % agr group I, 51 % group II and 10 % group III (P < 0.01) . The percentages of agr locus mutants and the three agr groups in different hospitals showed no significant differences (P > 0.05) . The percentage of agr group I S . epidermidis isolated from catheters and blood was higher than that isolated from the other clinical specimens . This is the first study to investigate the genetic polymorphism of agr in S . epidermidis in China . The mean percentage of agr locus mutants was 14.9 % (12 % in clinical specimens; 17.7 % in controls) . Genetic polymorphism of agr in S . epidermidis was linked to its pathogenicity; group I was associated with pathogenicity, while most isolates from healthy subjects were group II . The mechanism is to be investigated.

J Bacteriol, 2004 Jun, 186(11), 3374 - 83
Reconstitution of a staphylococcal plasmid-protein relaxation complex in vitro; Caryl JA et al.; The isolation of plasmid-protein relaxation complexes from bacteria is indicative of the plasmid nicking-closing equilibrium in vivo that serves to ready the plasmids for conjugal transfer . In pC221 and pC223, the components required for in vivo site- and strand-specific nicking at oriT are MobC and MobA . In order to investigate the minimal requirements for nicking in the absence of host-encoded factors, the reactions were reconstituted in vitro . Purified MobA and MobC, in the presence of Mg2+ or Mn2+, were found to nick at oriT with a concomitant phosphorylation-resistant modification at the 5' end of nic . The position of nic is consistent with that determined in vivo . MobA, MobC, and Mg2+ or Mn2+ therefore represent the minimal requirements for nicking activity . Cross-complementation analyses showed that the MobC proteins possess binding specificity for oriT DNA of either plasmid and are able to complement each other in the nicking reaction . Conversely, nicking by the MobA proteins is plasmid specific . This suggests the MobA proteins may encode the nicking specificity determinant.

J Bacteriol, 2004 Jun, 186(11), 3363 - 73
An accessory protein is required for relaxosome formation by small staphylococcal plasmids; Smith MC et al.; Mobilization of the staphylococcal plasmid pC221 requires at least one plasmid-encoded protein, MobA, in order to form a relaxosome . pC221 and closely related plasmids also possess an overlapping reading frame encoding a protein of 15 kDa, termed MobC . By completing the nucleotide sequence of plasmid pC223, we have found a further example of this small protein, and gene knockouts have shown that MobC is essential for relaxosome formation and plasmid mobilization in both pC221 and pC223 . Primer extension analysis has been used to identify the nic site in both of these plasmids, located upstream of the mobC gene in the sense strand . Although the sequence surrounding the nic site is highly conserved between pC221 and pC223, exchange of the oriT sequence between plasmids significantly reduces the extent of relaxation complex formation, suggesting that the Mob proteins are selective for their cognate plasmids in vivo.

Mol Immunol, 2004 May, 41(1), 1 - 8
Involvement of CTLA-4 in T-cell anergy induced by staphylococcal enterotoxin A in vitro; Xu GL et al.; Superantigens, like staphylococcal enterotoxin A (SEA), induce a strong proliferative response followed by clonal deletion of a substantial portion of defined V(beta) T-cells . The remaining cells display in vitro anergy . Anergy is a major mechanism to ensure antigen-specific tolerance in T-lymphocyte in the adult . Co-stimulatory molecules B7-1 (CD80)/B7-2 (CD86) and their counter-receptors CD28/CTLA-4 play pivotal roles in T-cell activation and immune regulation . While increasing data further suggested a role for CTLA-4 in regulating T-cell tolerance in vivo, the mechanism by which CTLA-4 influences T-lymphocyte tolerance is unclear . In the present study, we established an in vitro anergy model using superantigen SEA as the anergizing agents and examined CD3, CD28 and CTLA-4 expression of anergic T-cells in response to SEA rechallenge . It is found that anergic T-cell fails to produce the autocrine growth factor interleukin-2 (IL-2) upon stimulation, and addition of exogenous IL-2 can reverse the anergic state . Both TCR/CD3 complex and CD28 expression is not reduced in anergic cells during whole immune response, but the expression of CTLA-4 on the cell surface is enhanced dramatically in the late stages of an immune response . Using CTLA-4/B7-blocking agent, we found T-cell anergy was aborted and anergic T-cells restored the ability to proliferate and produce IL-2, suggesting that CTLA-4 may play a critical role in the induction of T-cell anergy.

J Appl Microbiol, 2004, 96(6), 1361 - 6
Identification and prevalence of an enterotoxin-related gene, se-int, in Staphylococcus intermedius isolates from dogs and pigeons; Futagawa-Saito K et al.; AIMS: To determine the prevalence of enterotoxin-producing Staphylococcus intermedius in dogs and pigeons . METHODS AND RESULTS: A total of 106 S . intermedius isolates from 44 dogs and 62 pigeons were tested for the production of enterotoxins A, B, C and D by reverse passive latex agglutination (RPLA) and for sec-canine by PCR . Only one isolate from dog was positive for SEC and sec-canine . Screening of sec-canine-negative strains by nested PCR led to the identification of a novel enterotoxin-related gene, se-int . SE-int showed a significant homology (59-61% identity) with SEC and (56.6% identity) SEB . All 44 isolates from dogs and five isolates (8.1%) from pigeons were se-int positive . CONCLUSIONS: While S . intermedius was isolated more frequently from pigeons than from dogs, se-int was more prevalent among the S . intermedius isolates from dogs, compared with the pigeon isolates . SIGNIFICANCE AND IMPACT OF THE STUDY: Further characterization of the se-int-positive S . intermedius strains should clarify their pathogenic potential including enterotoxigenicity and zoonotic transmissibility to human beings.

J Appl Microbiol, 2004, 96(6), 1265 - 70
A multiplex PCR for detection of genes encoding exfoliative toxins from Staphylococcus hyicus; Andresen LO et al.; AIMS: To develop a multiplex PCR for detection of genes encoding the exfoliative toxins ExhA, ExhB, ExhC and ExhD from Staphylococcus hyicus and to estimate the prevalence of exfoliative toxins among Staph . hyicus isolates from Danish pig herds with exudative epidermitis (EE) . METHODS AND RESULTS: A multiplex PCR employing specific primers for each of the genes encoding four different exfoliative toxins was developed and evaluated using a collection of Staph . hyicus with known toxin type and a number of other staphylococcal species . A total of 314 Staph . hyicus isolates from pigs with EE were screened by multiplex PCR and the combined results of the present and previous investigations showed that ExhA, ExhB, ExhC and ExhD was found in 20, 33, 18 and 22%, respectively, of 60 cases of EE investigated . CONCLUSIONS: This study has provided a new tool for detection of toxigenic Staph . hyicus and a more comprehensive picture of the prevalence of the Staph . hyicus exfoliative toxins in Danish pig herds . SIGNIFICANCE AND IMPACT OF THE STUDY: The multiplex PCR can be used in studies on the prevalence of toxigenic Staph . hyicus elucidating the epidemiology of EE in pigs . The multiplex PCR is currently being used for selection of Staph . hyicus isolates for production of autogenous vaccine.

Clin Pharmacokinet, 2004, 43(7), 417 - 40
Vancomycin: pharmacokinetics and administration regimens in neonates; de Hoog M et al.; This review describes the use of vancomycin in neonates over the last three decades . Given the relation of late-onset neonatal septicaemia to outcome and the increase in coagulase-negative staphylococcal infection as causative organism, vancomycin remains an important antibacterial in the neonatal intensive care unit . The pharmacokinetic behaviour of vancomycin in neonates can be adequately described by a one- or two-compartment model and is mainly determined by postconceptional age and renal function . In neonates, a patent ductus arteriosus as well as treatment with indomethacin or extracorporeal membrane oxygenation (ECMO) leads to an increase in volume of distribution and a decrease in clearance . Microbiological studies in vitro have shown that an increase in vancomycin concentrations above the minimum inhibitory concentration does not result in more effective killing . The microbiological and clinical efficacy of vancomycin in neonates has only been studied explicitly in a restricted number of patients . There are no definitive data relating serum concentrations to effect in this patient group . Vancomycin-related nephrotoxicity and ototoxicity in neonates is rare, and no clear relation to serum concentrations has been demonstrated . Based on the pharmacokinetic profile of vancomycin in neonates, several administration regimens have been constructed . Recent guidelines have suggested that dosage can be independent of gestational age or postconceptional age in neonates without renal failure . In patients with renal failure, therapy can be adequately tailored by using a regimen based on serum creatinine . The usefulness of routine monitoring of peak serum concentrations is doubtful based on the current literature . Recent research demonstrates a shift towards taking only routine trough serum concentrations in order to optimise efficacy . Patients with renal failure and other special subpopulations, such as patients exposed to ECMO or indomethacin, need to be monitored more closely.

Clin Diagn Lab Immunol, 2004 May, 11(3), 621 - 4
Caspase inhibitors attenuate superantigen-induced inflammatory cytokines, chemokines, and T-cell proliferation; Krakauer T; Proinflammatory cytokines mediate the toxic effect of superantigenic staphylococcal exotoxins (SE) . A pan-caspase inhibitor suppressed SE-stimulated T-cell proliferation and the production of cytokines and chemokines by human peripheral blood mononuclear cells . These data suggest that caspase inhibitors may represent a novel therapeutic modality for treating SE-induced toxic shock.

Cancer Biol Ther, 2004 Jul, 3(7), 660 - 6 Epub 2004 Jul 09.
In Vivo Tumor Co-Transfection with Superantigen and CD80 Induces Systemic Immunity without Tolerance and Prolongs Survival in Mice with Hepatocellular Carcinoma; Li ZS et al.; Background . Since transfection of established tumors with immunostimulatory genes can elicit antitumor immunity, we treat mouse HCC with in vivo transfection of superantigen SEA and/or costimulatory molecule CD80 and evaluated the safety and efficacy . Methods . Mice with HCC were treated with lipid-complexed plasmid DNA encoding staphylococcal enterotoxin A and/or CD80 . Then the mice were evaluated for tumor regression, systemic immunologic responses, survival times and treatment-associated toxicity . Results . Of all treated mice, the overall response rates (complete or partial remission) for SEA, CD80 and SEA/CD80 treated mice in this study were 65%, 60% and 75% separately, and were significantly higher than that of untreated mice . Most of the treat mice completed the therapy without any significant reaction . CTL activity increased with time of treatment and correlated temporally with an objective tumor response . Also our results indicated that local intratumoral expression of SEA did not lead to detectable deletion or anergy of SEA-reactive spleen T cells . Survival times for hepatoma mice in this study treated by intratumoral injection of SEA, CD80 and SEA/CD80 were prolonged significantly (P < 0.01) compared with the control mice.

Vet Pathol, 2004 May, 41(3), 299 - 301
Acute bacterial cystitis does not cause deoxyribonucleic acid damage detectable by the alkaline comet assay in urothelial cells of dogs; Alves A et al.; Considering the high incidence of dogs with acute bacterial cystitis (BC) and the relationship among inflammation, genotoxicity, and carcinogenesis, we conducted a case-control study comparing the frequency of deoxyribonucleic acid (DNA) lesions assessed by the comet assay between disease-free animals (13 males and 13 females) and cytology-confirmed cases of acute BC (12 males and 12 females), which was mainly caused by Staphylococcus sp . (40%) and Escherichia coli (35%) . The results show no increase in DNA damage in cells obtained by bladder washings and no influence of age, sex, and breed due to acute BC . In conclusion, DNA damage was seemingly not associated with the infection by specific bacteria.

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi, 2003 Jan, 19(1), 26 - 8
{The role of superantigen SEB-induced immunotolerance in the immune privileged site}; He QH et al.; AIM: To investigate if the immunological tolerance induced by superantigen staphylococcus enterotoxin B (SEB) takes part in the reaction at the immune privileged site . METHODS: Corneal transplantations were carried out using LEW rats as recipient and F344 rats as the donor . The recipient rats were randomly divided into 5 groups, receiving peribulbar SEB injection of a dosage of 30, 60, 90 or 120 microg/kg 7 days before and after operations . The negative control group received saline injection while the positive control group received glucocorticoid(GC), FK506, CsA or interleukin-1 receptor antagon ist (IL-1ra), respectively . The survival time and rejection index of corneal grafts within 30 days postoperation were examined and recorded using anti-rejection index method . The inflammatory cell infiltration of the recipient cornea and changes of local NK cells were examined by PE-conjugated anti-NK1.1 monoclonal antibody . RESULTS: The grafts in the group receiving injection of 120 microg/kg SEB survived 22 days longer than the negative control group . Their survival time is also significantly longer than the groups treated with FK506, CsA, GC or IL-1ra . The graft rejection index was 3.42+/-2.18 in the rats receiving 120 microg SEB/kg,but 6.58+/-3.15 in the negative control group (P<0.01) . The edema and new blood vessel indexes were also significantly reduced . The immunostaining showed that the number of NK cells increased following injection of SEB . CONCLUSION: Injection of SEB reduced the rejection to corneal transplants in rats, showing that the immune tolerance induced by superantigen SEB plays a role in the reaction at the immune privileged site.

J Vet Med B Infect Dis Vet Public Health, 2001 May, 48(4), 267 - 81
Chronic fatigue and immune dysfunction syndrome associated with Staphylococcus spp . bacteraemia responsive to thiacetarsamide sodium in eight birds of prey; Tarello W; Chronic fatigue and immune dysfunction syndrome (CFIDS) is a recognized human illness with zoonotic implications that is rarely described in animals . Eight birds of prey examined between 1992 and 1995 and sharing common symptoms (asthenia, inability to fly, poor appetite and emaciation) underwent laboratory tests revealing immunodeficiency, anaemia, high creatine kinase levels and low serum magnesium levels . Diagnosis of CFIDS was based upon these features . The effectiveness of an arsenic-based medication, thiacetarsamide sodium, administered intravenously for 2-3 days at low dosages (0.1 ml/kg/day) has been demonstrated by checks carried out 10, 20 and 30 days after therapy . The symptoms and the immune and haematological dysfunctions disappeared within 2-4 weeks of treatment . In all patients, micrococcus-like organisms found adhering to the outer surface of many red blood cells, had disappeared at post-treatment controls . Two of five blood cultures were positive for Staphylococcus spp . (S . intermedius and S . xilosus) . Consideration is given to the pharmacological activity of an arsenic-based drug in animal illnesses resembling CFIDS.

Clin Implant Dent Relat Res, 2003, 5(4), 226 - 32
Microbiologic diagnostics at titanium implants; Leonhardt A et al.; BACKGROUND: The microbiota found at periimplant lesions have been shown to contain putative periodontal pathogens as well as opportunistic species such as Staphylococcus spp, enterics, and Candida spp . Therefore, a microbiologic diagnosis may be of value as guidance before treatment of such lesions . PURPOSE: The aim of this study was to evaluate the prevalence of some putative pathogens associated with long-term followed-up cases using two different microbiologic procedures . MATERIALS AND METHODS: Fifteen subjects contributed with plaque samples from teeth and implants; these were analyzed with respect to 18 putative periimplant pathogens using cultural methods and a deoxyribonucleic acid DNA-DNA hybridization technique . RESULTS: The number of individuals positive for the analyzed pathogens was similar in samples taken from teeth and implants when analyzed with the DNA-DNA hybridization technique . When comparing detection frequency by culture procedure and by "checkerboard" technique at implants, the number of individuals positive for these species was lower with the traditional culture technique than with the checkerboard analyses . Using a higher cutoff point (> or = 4) with the checkerboard technique, the number of positive individuals was generally lower than that found with the culture technique . When comparing the techniques on an implant site level, the prevalence obtained by culture was lower for all analyzed species . If the specific species were present in the samples analyzed by the checkerboard technique, they were present only in every second sample analyzed with the culture technique . The high specificity values showed that if the checkerboard technique did not detect any Porphyromonas gingivalis, Prevotella intermedia, Actinobacillus actinomycetemcomitans, or Fusobacterium nucleatum, the bacteria were also undetectable by the culture technique . The two methods therefore did not overlap but did supplement each other . CONCLUSIONS: Based on the current results it is recommended that the technique used when analyzing microbiota around titanium implants should be a combination of the two protocols mentioned as they seem to give the most comprehensive outcome when used together.

Laryngoscope, 2004 May, 114(5), 887 - 92
Microbial superantigens induce glucocorticoid receptor beta and steroid resistance in a nasal explant model; Fakhri S et al.; OBJECTIVE: To study the role of superantigen (SAg) in inducing glucocorticoid (GC) receptor beta and steroid resistance in an explant model of nasal tissue . METHODS: Nasal tissue was obtained from inferior turbinates of controls and ragweed (RW)-sensitive patients . Tissue samples were incubated with SAg of staphylococcal enterotoxin B . In addition, tissue samples from RW-sensitive patients were incubated with RW allergen in the presence and absence of both SAg and dexamethasone (DEX) . The expression of GC receptor beta was assessed by immunocytochemistry . The expression of interleukin (IL)-2 and IL-4 mRNA was assessed by in situ hybridization . RESULTS: SAg induced an increase in the expression of GC receptor beta in atopic tissue and to a lesser extent in nonatopic tissue . The most significant induction of GC receptor beta was observed in response to SAg and RW in atopic tissue . Stimulation of atopic tissue with RW alone and SAg alone induced IL-4 and IL-2 mRNA, respectively . Incubation of atopic tissue with both SAg and RW induced both IL-2 and IL-4 mRNA . The increase in IL-4 mRNA expression was blunted by the addition of DEX to atopic tissue stimulated with RW alone but not to tissue stimulated by both RW and SAg . CONCLUSION: Our results demonstrate that SAgs induce steroid resistance in atopic nasal explant tissue by up-regulating the expression of GC receptor beta . Furthermore, we have shown that the up-regulation of GC receptor beta is a local event that is associated with the coexpression of IL-2 and IL-4 mRNA.

J Infect Dis, 2004 May 15, 189(10), 1892 - 6 Epub 2004 Apr 26.
Human peripheral gammadelta T cells potentiate the early proinflammatory cytokine response to staphylococcal toxic shock syndrome toxin-1; Kalyan S et al.; Toxic shock syndrome toxin (TSST)-1 is a superantigen known to profoundly induce proinflammatory cytokines by activation of V beta -specific alpha beta T cells, but its effect on gamma delta T cells, which normally constitute 1%-5% of peripheral blood mononuclear cells (PBMCs), is unclear . Here, we demonstrate that TSST-1 induced significantly higher levels of interferon (IFN)- gamma, tumor necrosis factor (TNF)- alpha, and interleukin (IL)-2, and a lower level of IL-10 in human PBMCs when the gamma delta subpopulation has been primed by isopentylpyrophosphate, compared with that in control PBMCs . Furthermore, depletion of the gamma delta subpopulation completely abrogated this effect . Thus, peripheral gamma delta T cells markedly modulate both the proinflammatory and anti-inflammatory cytokine responses of TSST-1.

Eur J Vasc Endovasc Surg, 2004 Jun, 27(6), 603 - 7
RNAIII-inhibiting peptide and/or nisin inhibit experimental vascular graft infection with methicillin-susceptible and methicillin-resistant Staphylococcus epidermidis; Ghiselli R et al.; OBJECTIVE: To investigate the efficacy of RNAIII-inhibiting peptide (RIP) and nisin as prophylactic agents in a rat model of vascular graft infection . DESIGN: Prospective, randomized, controlled animal study . MATERIALS: Two hundred and twenty adult male Wistar rats . Staphylococcus epidermidis ATCC 12228 and one clinical isolate of methicillin-resistant S . epidermidis . Drugs: RIP, nisin and rifampin . METHODS: Graft infections were established in the dorsal subcutaneous tissue by implantation of 1 cm(2) sterile Dacron grafts, followed by topical bacterial inoculation: grafts were retrieved at 7 days . The study included a control group (without inoculation) and two series composed of five groups for each staphylococcal strain: one contaminated group that did not receive any antibiotic prophylaxis, three contaminated groups that received grafts soaked with 10 mg/l RIP, 10 mg/l nisin, 10 mg/l rifampin, or RIP+nisin . The main outcome measure was the extent of bacterial at graft harvest . RESULTS: The bacterial counts for methicillin-resistant S . epidermidis on explanted grafts were 6.1+/-2.8x10(2), 7.8+/-3.0x10(3) and 5.5+/-2.9x10(4) for RIP, nisin and rifampin, respectively . RIP and nisin used in combination reduced the bacterial count to <10 . The results for S . epidermidis were similar . CONCLUSIONS: RIP and nisin could be used in combination to coat medical devices to prevent drug resistant S . epidermidis infections.

Scand J Infect Dis, 2004, 36(3), 209 - 12
Cold bone defect on granulocytes labelled with technetium-99m-HMPAO scintigraphy: significance and usefulness for diagnosis and follow-up of osteoarticular infections; Galperine T et al.; We wished to determine the frequency and significance of cold bone defect on granulocytes labelled with technetium-99-m-hexamethylpropyleneamine oxime (99mTc-HMPAO-PMN) in non-spinal bone infection . Cold bone defect was investigated as part of a retrospective review during a 2-y period . Patients who had possible osteoarticular infection underwent bone scintigraphy combined with 99mTc-HMPAO-PMN for diagnosis and follow-up . Osteomyelitis was confirmed by isolation of the responsible pathogen . Among 210 patients who had possible infection, 17 (8%) demonstrated a cold bone defect . The site of cold bone defect was for all patients the hip . All 17 patients had proven bacterial orthopaedic hardware-related infection . The single causative micro-organism was staphylococcus . Whatever the outcome, cold bone defect was constant regardless of follow-up equal to or longer than 18 months . These data suggest that this uncommon scintigraphic pattern is an indication of an infectious process similar to increased uptake.

J Laryngol Otol, 2004 Apr, 118(4), 260 - 2
Microbiology of cerumen in patients with recurrent otitis externa and cases with open mastoidectomy cavities; Pata YS et al.; This study investigated the common flora of human cerumen in patients with recurrent otitis externa, and subjects who had been operated on and had an open mastoidectomy cavity from chronic otitis media . Cerumen samples were collected from three groups; group A (n = 20) consisted of patients with recurrent otitis externa, group B (n = 20) consisted of patients with an open cavity and group C (n = 30) consisted of healthy subjects . The mean of the microbial count was 3.4 x 10(4) in group A, 3.08 x 10(4) in group B and 2.48 x 10(4) in group C . The most commonly isolated microorganism from the three groups was Staphylococcus epidermidis . No growth was observed in five cases (25 per cent) in group A and in three cases (10 per cent) in group C . In group B antimicrobial growth was observed in all samples . In 46 (65 per cent) of the cerumen samples, the isolates were monomicrobial and 24 (35 per cent) of the cerumen samples were polymicrobial . The isolates were polymicrobial in 65 per cent of group A, 20 per cent in group B and 23.3 per cent in group C . In the process of investigating the microbial flora of cerumen in all the three groups, microbial growth was observed from all the samples from patients with an open cavity, unlike the other groups, and it was determined that the group with recurrent external otitis had the most abundant microbial flora.

J Perinatol, 2004 May, 24(5), 324 - 6
Mixed infection with unusual fungi and staphylococcal species in two extremely premature neonates; Maheshwari A et al.; Systemic fungal infections are being recognized with increasing frequency in extremely premature neonates . We report two such infants with late-onset mixed infection with Staphylococcal species and unusual fungi . These cases are of interest in view of recent reports on the interaction of Staphylococcal cell wall components and neutrophils, as damaged skin sites could form a nidus and portal of entry for saprophytic fungal pathogens . It is also important to consider fungal infection as a possibility in sick premature infants with necrotic skin lesions even when the systemic signs have an alternative explanation with ongoing bacteremia.

Indian J Med Res, 2004 Mar, 119(3), 107 - 9
Effect of growth medium on hydrophobicity of Staphylococcus epidermidis; Das SC et al.; The present study was undertaken to evaluate the effect of growth medium on expression of hydrophobicity of Staphylococcus epidermidis . A total of 24 hydrophobic isolates of S . epidermidis, determined by n-hexadecane adherence assay (HAA) earlier were included . Five different growth media: horse blood agar (HBA), brain heart infusion agar (BHIA), brain heart infusion broth (BHIB), tryptic soy broth (TSB) and proteose peptone broth (PPB) were used . All 24 isolates exhibited the reproducible hydrophobicity when grown on HBA; however, 20 (83.33%), 19 (79.16%), 15 (62.50%) and 13 (54.16%) isolates were found to be hydrophobic when grown in BHIA, BHIB, TSB and PPB, respectively . HBA was found to be the most suitable medium for detection of hydrophobicity of S . epidermidis followed by BHIA or BHIB.

J Clin Immunol, 2004 May, 24(3), 294 - 301
A novel approach for cancer immunotherapy: tumor cells with anchored superantigen SEA generate effective antitumor immunity; Ma W et al.; Murine B16 melanoma cell line is poorly immunogenic and highly aggressive . We recently reported that the transmembrane staphylococcal enterotoxin A (TM-SEA) anchors onto B16 cells and stimulates lymphocyte proliferation . The purpose of the study was to investigate whether vaccination with B16 cells bearing membrane-anchored TM-SEA fusion protein could cause tumor-specific immunity . Mice in the therapeutic vaccination group received B16 tumor inoculations, followed by treatment with B16-TM-SEA vaccine or control vaccines . Mice in the prophylactic vaccination group were given B16-TM-SEA vaccine or control vaccines, followed by challenge with wild type B16 or control EL4 cells . Significant tumor growth inhibition, prolongation of survival, and marked augmentation of NK and CTL activities were observed in mice which received B16-TM-SEA vaccine as compared to controls . Overall, our results suggest that the TM-SEA cellular vaccine is a novel and effective strategy for cancer immunotherapy.

Ann Pharmacother, 2004 Jun, 38(6), 986 - 8 Epub 2004 Apr 27.
Successful treatment of Staphylococcus epidermidis hip prosthesis infection with oral linezolid; Mogenet I et al.; OBJECTIVE: To report a case of Staphylococcus epidermidis infection of a hip prosthesis successfully treated with oral linezolid . CASE SUMMARY: A 46-year-old woman developed methicillin-resistant S . epidermidis (MRSE) infection of her prosthetic hip . She received oral linezolid 600 mg twice daily for one month; after that time, the biological inflammatory markers returned to normal . DISCUSSION: One of the most serious complications of arthroplasty is joint prosthesis infection . It is mainly caused by gram-positive bacteria, in particular those of the genus staphylococcus . The increasing prevalence of gram-positive cocci that are resistant to antimicrobial agents has complicated the treatment of serious infections . CONCLUSIONS: Oral linezolid appears to be an effective and well-tolerated treatment option for hip prosthesis infections due to MRSE.

Ann Otol Rhinol Laryngol, 2004 Apr, 113(4), 310 - 2
Clinical ototoxicity of teicoplanin; Bonnet RM et al.; No strong evidence of ototoxicity of teicoplanin can be found in the literature, possibly because of conservative definitions of ototoxicity . We performed audiometry over time to compare the ototoxicity of teicoplanin with that of cloxacillin as a non-ototoxic standard . The data were analyzed with a linear mixed-effects model . The hearing thresholds of 12 patients who were treated with teicoplanin for severe staphylococcal infections showed a slight but significant increase over time, whereas the thresholds of 5 patients treated with cloxacillin decreased significantly during treatment . This improvement in hearing with cloxacillin may be attributed to improvement of the clinical condition . This outcome implies that previous reports that suggest a lack of ototoxicity of teicoplanin potentially underestimate the risk and should be interpreted accordingly.

World J Gastroenterol, 2004 May 1, 10(9), 1369 - 71
Diagnosis of intestinal acariasis with avidin-biotin system enzyme-linked immunosorbent assay; Zhang RB et al.; AIM: To explore the value of avidin-biotin system enzyme-linked immunosorbent assay (ABC-ELISA) in diagnosis of intestinal acariasis . METHODS: Mite-specific IgG levels in serum of 48 patients with intestinal acariasis were measured with ABC-ELISA . The sensitivity of this method was compared with that of staphylococcal protein A enzyme-linked immunosorbent assay (SPA-ELISA) . RESULTS: The positive rate of mite-specific IgG detected with ABC-ELISA and SPA-ELISA was 89.58% (43/48) and 56.25% (27/48), respectively . The positive rate with ABC-ELISA was statistically higher than that with SPA-ELISA (chi2=13.50, P<0.01) . CONCLUSION: ABC-ELISA is an effective method for the diagnosis of intestinal acariasis.

J Clin Anesth, 2004 Mar, 16(2), 92 - 7
Bacterial contamination of epidural catheters: microbiological examination of 502 epidural catheters used for postoperative analgesia; Steffen P et al.; STUDY OBJECTIVE: To investigate the frequency of bacterial colonization of epidural catheters used for postoperative pain treatment longer than 24 hours in abdominal, thoracic, or trauma surgery patients . DESIGN: Retrospective study . SETTING: Intermediate care facility and general ward of a university hospital . PATIENTS: 502 patients who received epidural catheters after abdominal, thoracic, or vascular surgery at our institution from January 1996 to December 2000 . INTERVENTIONS: Placement of an epidural catheter, which was used for postoperative pain treatment, for more than 24 hours . The puncture site dressing included saturation each day with povidone-iodine . MEASUREMENTS AND MAIN RESULTS: Microbiologic monitoring of epidural catheter tips and daily examination of puncture sites with regard to signs of inflammation took place . Four times daily patients were examined to check adequacy of pain treatment and neurologic deficits . Catheter tip cultures were positive in 29 patients (5.8%) . Staphylococcus epidermidis was isolated in 22 cases (76%) . No case of spinal epidural abscess was observed within 6 months after epidural catheterization . The average catheterization time was 5 days (quartile range: 4 to 6 days) . CONCLUSIONS: Meticulous management ensures a relatively low level of bacterial contamination in epidural catheters applied for postoperative pain treatment greater than 5 days . Contamination rarely leads to spinal epidural infection.

FEMS Microbiol Lett, 2004 May 1, 234(1), 155 - 61
A von Willebrand factor-binding protein from Staphylococcus lugdunensis; Nilsson M et al.; In the present study, a phage display library covering the genome of Staphylococcus lugdunensis, was affinity-selected against von Willebrand factor (vWf) . This led to the identification of a gene, vwbl, encoding a putative cell surface protein of 2060 amino acids, denoted vWbl . The deduced protein has an overall organisation typical of staphylococcal cell surface proteins, with an N-terminal signal peptide, and a C-terminal cell wall sorting signal . The vWf-binding part is located in repetitive domains and antibodies against vWbl or vWf can inhibit the binding . Southern blot analysis showed that vwbl was present in the 12 S . lugdunensis strains tested.

Kansenshogaku Zasshi, 2004 Jan, 78(1), 46 - 53
{Drug sensitivity and teicoplanin resistance of coagulase-negative Staphylococcus isolated from blood culture}; Nakamura T et al.; Drug sensitivity and teicoplanin (TEIC) resistance of coagulase-negative Staphylococcus spp (CNS) isolated from blood culture in 10 years between 1992 and 2001 were investigated, and epidemiological analysis was performed using the survey of clinical backgrounds . Regarding the resistance rates of CNS to various drugs, no isolate was resistant to vancomycin (VCM), arbekacin (ABK), and linezolid (LZD), but the minimum inhibitory concentration (MIC) of TEIC was 16 micrograms/ml or higher in 10.3% of isolates . The isolates include 56 S . epidermidis strains, 1 S . capitis strain and 4 S . haemolyticus strains . As for other drugs, the minocycline (MINO) resistance rate was relatively low (6.1%), and methicillin-resistant CNS (MRCNS) accounted for 67.1% . The resistance rate was high for many drugs between 1995 and 1997 . MIC50/90 of drugs considered to be anti-Staphylococcus were 1/2 microgram/ml (VCM), 1/16 microgram/ml (TEIC), 0.25/1 microgram/ml (ABK) and 1/2 microgram/ml (LZD), showing that both MIC 50 and MIC90 of ABK were the lowest . Regarding cross-resistance to TEIC, the resistant group (MIC: 16 micrograms/ml or higher) exhibited cross-resistance to all drugs excluding MINO . Particularly, 90.2% of the TEIC resistant group was MRCNS . TEIC-resistant strain was most frequently (30.8%) detected in the pediatric ward . In comparison of clinical backgrounds of the TEIC-resistant and TEIC-sensitive groups, there were no significant differences in age, use of IVH, or prognosis, but a significant difference in detection of TEIC-resistant strains was noted in patients treated with the 4 th cepharosporins and carbapenems (p < 0.01) . This survey clarified the presence of TEIC-resistant CNS strains . We will continue the survey to monitor resistance conversion . Investigation of the involvement of TEIC-resistant CNS strains in hospital infection and therapeutic course is also necessary.

Phytother Res, 2004 Mar, 18(3), 212 - 23
Biological activity of barbados cherry (acerola fruits, fruit of Malpighia emarginata DC) extracts and fractions; Motohashi N et al.; Fractionation of barbados cherry (acerola fruit, a fruit of Malpighia emarginata DC.) extracts were performed by organic solvent extractions and column chromatographies, using two extraction methods . Higher cytotoxic activity was concentrated in fractions A4 and A6 (acetone extract), and H3 and HE3 (hexane extract) . These four fractions showed higher cytotoxic activity against tumor cell lines such as human oral squamous cell carcinoma (HSC-2) and human submandibular gland carcinoma (HSG), when compared with that against normal cells such as human periodontal ligament fibroblasts (HPLF) and human gingival fibroblasts (HGF) . HE2 (hexane extract), AE2 (ethyl acetate extract), AE3, AE4, AE5, A8, A9 and A10 showed some relatively higher anti-bacterial activity on the Gram-positive Staphylococcus epidermidis ATCC 1228 but were ineffective on the representative Gram-negative species E . coli and Ps . aeruginosa . The fractions were inactive against Helicobacter pylori, two representative Candida species, and human immunodeficiency virus (HIV) . H3, H4 and HE3, which displayed higher tumor-specific cytotoxicity also showed higher multidrug resistance (MDR) reversal activity, than (+/-)-verapamil as positive control . ESR spectroscopy shows that the radical-mediated oxidation is not involved in the induction of tumor-specific cytotoxic activity . The tumor specific cytotoxic activity and MDR reversal activity of barbados cherry may suggest its possible application for cancer therapy .

Infect Immun, 2004 May, 72(5), 3081 - 3
The fibrinogen binding protein of Staphylococcus epidermidis is a target for opsonic antibodies; Rennermalm A et al.; Antibodies against the fibrinogen binding protein (Fbe) of Staphylococcus epidermidis significantly increased macrophage phagocytosis . Antibodies against autolysin E were opsonic but to a lesser extent . Antibodies against a novel, putatively surface-located antigen were unable to enhance phagocytosis . The severity of systemic infection of mice with S . epidermidis was reduced if the bacteria were preopsonized with anti-Fbe prior to administration . Fbe is thus a strong candidate for protein vaccination against S . epidermidis infection, and antibodies against Fbe can be used to prevent or treat infections caused by S . epidermidis.

J Allergy Clin Immunol, 2004 Apr, 113(4), 710 - 6
High IFN-gamma production by CD8+ T cells and early sensitization among infants at high risk of atopy; Rowe J et al.; BACKGROUND: High genetic risk (HR) of atopy among unstratified populations of infants is associated with attenuated IFN-gamma responses . However, the role of IFN-gamma in progression from HR status to active disease is less clear . OBJECTIVE: To identify immune function markers in neonates with HR that are associated with positive atopic outcomes at 2 years . METHODS: Cord blood mononuclear cells (CBMCs) were collected from 175 children with HR and cryopreserved . The children were assessed for atopy by skin prick at 0.5 and 2 years . CBMCs were thawed and stimulated with allergens and mitogens PHA and staphylococcal enterotoxin B (SEB), and cytokine responses were determined . RESULTS: No correlations were observed between allergen-specific CBMC responses and atopic outcomes . In contrast, sensitization was strongly associated with polyclonal IFN-gamma responses to both PHA (P=.002) and SEB (P=.005), and also with SEB-induced IL-5 (P =.05), IL-10 (P =.02), and IL-13 (P =.01) . Logistic regression analysis identified elevated PHA-induced IFN-gamma and SEB-induced IL-13 responses as the strongest independent predictors of atopy development . Cell separation studies confirmed CD8+ T cells as the source of approximately 90% of IFN-gamma production . CONCLUSIONS: IFN-gamma produced by CD8+ T cells may synergize with T(H)2 cytokines in driving atopy development in children with HR.

Ann Pharmacother, 2004 Jun, 38(6), 989 - 91 Epub 2004 Apr 20.
Treatment of vancomycin-resistant enterococcus with quinupristin/dalfopristin and high-dose ampicillin; Bethea JA et al.; OBJECTIVE: To report the successful treatment of vancomycin-resistant Enterococcus (VRE) bacteremia using the combination of quinupristin/dalfopristin and high-dose ampicillin . CASE SUMMARY: A 38-year-old African American woman with relapsed acute myeloid leukemia and neutropenic fever developed VRE bacteremia following 3 successive courses of vancomycin for methicillin-resistant staphylococcal infections . Treatment with linezolid was initiated; however, after 9 days of therapy, blood cultures continued to reveal VRE and the patient became febrile . The patient was subsequently switched to quinupristin/dalfopristin and high-dose ampicillin . The fever resolved and all subsequent blood cultures were negative after the initiation of combination therapy . DISCUSSION: The emergence of VRE infections presents a treatment challenge in immunocompromised patients . When treating VRE infections in this patient population, the effectiveness of linezolid and quinupristin/dalfopristin is limited by their bacteriostatic activity when used as monotherapy . Recent in vitro data suggest synergistic activity with quinupristin/dalfopristin when used in combination with other antimicrobials in selected isolates of VRE . CONCLUSIONS: Persistent VRE bacteremia was successfully treated in this neutropenic patient using the combination of high-dose ampicillin and quinupristin/dalfopristin . Case reports and in vitro data suggest that concomitant therapy with high-dose ampicillin may be an effective treatment alternative for VRE infections not responding to standard therapy.

J Immunol, 2004 May 1, 172(9), 5450 - 5
Lymphocyte activation gene-3 (CD223) regulates the size of the expanding T cell population following antigen activation in vivo; Workman CJ et al.; Lymphocyte activation gene-3 (LAG-3) is a CD4-related, activation-induced cell surface molecule that binds to MHC class II with high affinity . In this study, we used four experimental systems to reevaluate previous suggestions that LAG-3(-/-) mice had no T cell defect . First, LAG-3(-/-) T cells exhibited a delay in cell cycle arrest following in vivo stimulation with the superantigen staphylococcal enterotoxin B resulting in increased T cell expansion and splenomegaly . Second, increased T cell expansion was also observed in adoptive recipients of LAG-3(-/-) OT-II TCR transgenic T cells following in vivo Ag stimulation . Third, infection of LAG-3(-/-) mice with Sendai virus resulted in increased numbers of memory CD4(+) and CD8(+) T cells . Fourth, CD4(+) T cells exhibited a delayed expansion in LAG-3(-/-) mice infected with murine gammaherpesvirus . In summary, these data suggest that LAG-3 negatively regulates T cell expansion and controls the size of the memory T cell pool.

Aust Vet J, 2003 Aug, 81(8), 472 - 5
Metaphyseal osteomyelitis in an immature Abyssinian cat; Bradley WA; Metaphyseal osteomyelitis is a rarely described condition in small animals, especially the cat . Infection, most commonly Staphylococcus sp, is considered to occur in the metaphyseal region of the immature animal due to vascular anomalies that predispose to the haematogenous seeding of bacteria in this area . There is also speculation that the characteristics of the bacteria that allow them to adhere to cartilage matrix, rather than vascular linings and erythrocytes, may provide an advantage for colonisation in the metaphysis, resulting in infection . This case describes the successful management of a case of distal radial metaphyseal osteomyelitis in an immature cat using surgical intervention and antibacterial therapy.

Ann Pharmacother, 2004 Jun, 38(6), 936 - 41 Epub 2004 Apr 14.
Extended-interval aminoglycoside dosing for treatment of enterococcal and staphylococcal osteomyelitis; Graham AC et al.; BACKGROUND: Gram-positive osteomyelitis requires long-term antibiotic therapy, much of which is often administered in the outpatient setting . Historically, synergistic aminoglycoside use in these infections requires multiple daily doses, which can be inconvenient . Data regarding extended-interval aminoglycoside dosing (EIAD), also known as once-daily dosing, in this setting are lacking . OBJECTIVE: To evaluate the safety and efficacy of EIAD in the treatment of gram-positive osteomyelitis . METHODS: Retrospective chart review of adult patients treated for documented, gram-positive osteomyelitis with EIAD at the University of New Mexico Home IV Antibiotic Clinic was conducted . The patients' medical records were reviewed by an infectious diseases clinical pharmacist . Clinical and microbiologic outcomes and the incidence of nephrotoxicity and ototoxicity were the main outcome measures . RESULTS: Fifteen patients (16 events) were included . Enterococcus spp . was the most common organism isolated . Nine patients had infected equipment or devices; 6 of these had removal of these devices in conjunction with antibiotic therapy . The median duration of antibiotic therapy was 6 weeks (range 6-31) . The median duration of aminoglycoside therapy was 28 days (range 6-43) . Seven patients developed nephrotoxicity, 5 of whom received an aminoglycoside in combination with vancomycin . Male patients had a higher risk of developing nephrotoxicity compared with females (p = 0.04) . The mean +/- SD duration of EIAD before the development of nephrotoxicity was 34 +/- 8 days . Clinical cure was achieved in 12 (75%) patients . Three patients achieved clinical cure without hardware removal . CONCLUSIONS: Most of the patients with gram-positive osteomyelitis were successfully managed with EIAD . However, nephrotoxicity developed in a high proportion of patients and was likely related to prolonged aminoglycoside use.

Aust Vet J, 2003 Oct, 81(10), 608 - 11
Metaphyseal osteomyelitis in an immature Abyssinian cat; Bradley WA; Metaphyseal osteomyelitis is a rarely described condition in small animals, especially the cat . Infection, most commonly Staphylococcus sp, is considered to occur in the metaphyseal region of the immature animal due to vascular anomalies that predispose to the haematogenous seeding of bacteria in this area . There is also speculation that the characteristics of the bacteria that allow them to adhere to cartilage matrix, rather than vascular linings and erythrocytes, may provide an advantage for colonisation in the metaphysis, resulting in infection . This case describes the successful management of a case of distal radial metaphyseal osteomyelitis in an immature cat using surgical intervention and antibacterial therapy.

J Appl Microbiol, 2004, 96(5), 1185 - 93
Catabolism of leucine to branched-chain fatty acids in Staphylococcus xylosus; Beck HC et al.; AIMS: Staphylococcus xylosus is an important starter culture in the production of flavours from the branched-chain amino acids leucine, valine and isoleucine in fermented meat products . The sensorially most important flavour compounds are the branched-chain aldehydes and acids derived from the corresponding amino acids and this paper intends to perspectivate these flavour compounds in the context of leucine metabolism . METHODS AND RESULTS: GC and GC/MS analysis combined with stable isotope labelling was used to study leucine catabolism . This amino acid together with valine and isoleucine was used as precursors for the production of branched-chain fatty acids for cell membrane biosynthesis during growth . A 83.3% of the cellular fatty acids were branched . The dominating fatty acid was anteiso-C(15:0) that constituted 55% of the fatty acids . A pyridoxal 5'-phosphate and alpha-ketoacid dependent reaction catalysed the deamination of leucine, valine and isoleucine into their corresponding alpha-ketoacids . As alpha-amino group acceptor alpha-keto-beta-methylvaleric acid and alpha-ketoisovaleric acid was much more efficient than alpha-ketoglutarate . The sensorially and metabolic key intermediate on the pathway to the branched-chain fatty acids, 3-methylbutanoic acid was produced from leucine at the onset of the stationary growth phase and then, when the growth medium became scarce in leucine, from the oxidation of glucose via pyruvate . CONCLUSIONS: This paper demonstrates that the sensorially important branched-chain aldehydes and acids are important intermediates on the metabolic route leading to branched-chain fatty acids for cell membrane biosynthesis . SIGNIFICANCE AND IMPACT OF THE STUDY: The metabolic information obtained is extremely important in connection with a future biotechnological design of starter cultures for production of fermented meat.

Cornea, 2004 Mar, 23(2), 136 - 42
Aerobic bacterial conjunctival flora in diabetic patients; Martins EN et al.; OBJECTIVE: To study the aerobic conjunctival flora of diabetic patients and its relation to the presence and level of diabetic retinopathy and the duration of the disease . METHODS: One hundred three patients from the diabetic retinopathy screening program of the Federal University of Sao Paulo with no evidence of ocular surface disease were included . The diabetic patient cohort was compared with 60 nondiabetic subjects . All patients underwent slit-lamp evaluation, conjunctival scrapings, and indirect ophthalmoscopy . RESULTS: The frequency of positive conjunctival cultures was significantly higher in the diabetic group (94.18%) than in the nondiabetic group (73.33%) . Among diabetic patients, a significantly higher frequency of positive cultures was detected in those with diabetic retinopathy than in those without retinopathy . Neither the duration of the diabetes nor the hypoglycemic therapy correlated with the culture results . Coagulase-negative Staphylococcus was the most common microorganism isolated, and its identification was more frequent in patients with retinopathy than in those without diabetic retinopathy . CONCLUSION: Diabetic patients have a significantly higher number of positive conjunctival cultures . The presence of diabetic retinopathy was correlated with an increase in positive cultures and a higher proportion of coagulase-negative Staphylococcus.

Anticancer Drugs, 2004 Feb, 15(2), 113 - 8
SR31747A: a peripheral sigma ligand with potent antitumor activities; Casellas P et al.; SR31747A is currently being evaluated in phase IIb clinical trials for prostate cancer treatment . The molecule is a peripheral sigma ligand that binds four proteins in human cells, i.e . SRBP-1, sigma-2, HSI and its relative SRBP-2 . SR31747A is a dual agent with both immunomodulatory and antiproliferative activities . The molecule blocks proliferation of human and mouse lymphocytes, modulates the expression of pro- and anti-inflammatory cytokines, and was shown to protect animals in vivo against acute and chronic inflammatory conditions such as acute graft-versus-host reaction, lethality induced by staphylococcal enterotoxin B and lipopolysaccharide or rheumatoid arthritis . Besides these immunomodulatory activities, the molecule also inhibits the proliferation of various tumor cell lines in vitro in a time- and concentration-dependent manner . In vivo, SR31747A has potent antitumoral activity as demonstrated against mammary and prostatic tumoral cell lines injected into nude mice, where both tumor incidence and growth were decreased by more than 40% following daily SR31747A treatment at 25 mg/kg i.p . The recent literature on SR31747A in cancer is reviewed here . We focus specifically on preclinical data obtained in vivo and on studies aimed at deciphering the mode of action of the molecule.

Scand J Gastroenterol, 2004 Mar, 39(3), 238 - 45
Increased expression of TCR vbeta5.1 and 8 in mucosal T-cell lines cultured from patients with Crohn disease; Kelsen J et al.; BACKGROUND: Characterization of the T-cell receptor variable beta chain (Vbeta) repertoire in inflamed mucosa has been used to identify disease-relevant T-cell populations and antigens in Crohn disease (CD) . In vitro expansion of mucosal T cells may reveal changes in Vbeta repertoire not apparent in fresh isolates and we aimed to identify Vbeta subpopulations implicated in Crohn disease . METHODS: In vivo activated mucosal T cells were cultivated using IL-2 and IL-4 from biopsies of whole colonic mucosa without use of Vbeta-modifying exogenous antigen or feeder cells . The Vbeta gene expression in mucosal T-cell cultures was determined in 30 patients with CD and 12 healthy controls using reverse transcriptase polymerase reaction (RT-PCR) covering all 23 functional Vbeta families and the Vbeta receptor prevalence was evaluated by flow cytometry in selected cultures . RESULTS: Early T-cell cultures from both CD patients and healthy controls showed a polyclonal Vbeta gene expression that narrowed during culture, which in CD cultures led to a significant over-expression of the Vbeta5.1 (P = 0.04) and Vbeta8 gene segments (P = 0.03) . Together with Vbeta6 and Vbeta18, these Vbeta chains form a pattern of staphylococcal enterotoxin type E (SEE) responsive Vbeta chains, also over-expressed in CD cultures (P = 0.02) . Further in vitro stimulation of CD cultures with SEE caused expansion of Vbeta8 receptor positive cells together with a proinflammatory cytokine response . CONCLUSIONS: CD may be associated with (super)antigen-specific Vbeta subpopulations selected during long-term cultivation of mucosal biopsies from inflamed colon.

Arch Inst Pasteur Tunis, 2002, 79(1-4), 19 - 26
{In vivo and in vitro protection against lethal activity of Buthus occitanus tunetanus venom with a recombinant protein}; Benkhadir K et al.; We report the use of recombinant scorpion toxin in the form of fusion protein as antigen for mice immunisation . The aim is to produce protective antisera against lethal activity of the venom from Tunisian scorpion Buthus occitanus tunetanus, responsible for several annually reported human cases of scorpion stings . The gene encoding Bot III (the most toxic alpha toxin of Buthus occitanus tunetanus) was fused to the sequence encoding synthetic ZZ domains of staphylococcal protein A . The construct ZZ-Bot III was expressed in the periplasm of E . coli as a fusion protein and purified by affinity chromatography . The recombinant fusion protein was characterized and used as antigen to generate antibodies in mice . The antibodies against the recombinant protein neutralize the toxic venom (10 LD50/ml) and also confer protection for immunized mice against antigenically related mammal toxins.

J Immunol, 2004 Apr 15, 172(8), 4883 - 92
Burn injury initiates a shift in superantigen-induced T cell responses and host survival; Zang Y et al.; Severe injury induces a temporal shift in immune reactivity that can cause serious complications or even death . We previously reported that mice exposed to bacterial superantigen (SAg) early after injury undergo a strong SAg response with lethal consequences . This study compares the early and late effects of burn injury on SAg reactivity in vivo to establish how injury influences adaptive immune responses . We found that mice challenged with ordinarily sublethal doses of staphylococcal enterotoxin A or staphylococcal enterotoxin B at 1 day after burn injury exhibited high mortality, whereas no mortality occurred at 7 days after injury . This shift in mortality correlated with higher Th2-type cytokines (IL-4 and IL-10) being expressed by CD4(+) and CD8(+) T cells from burn as opposed to sham mice at 7 days after injury . Lymph node cells from burn-injured mice also produced higher levels of Th2-type cytokines at 7 days after injury . The results of cell-mixing studies using CD4(+) and CD8(+) T cells mixed with APCs from sham or burn mice suggested that changes in both T cells and APCs are involved in the altered SAg response . Finally, the biological significance of altered SAg reactivity following injury was shown by demonstrating that blocking IL-10 activity in vivo caused higher SAg-induced mortality at 7 days after injury . These findings support the idea that injury promotes a Th2-type shift in adaptive immune reactivity . Although prior studies link this counterinflammatory-type response to lowered resistance to infection, the present results suggest it may sometimes benefit the injured host.

J Biotechnol, 2004 Apr 29, 109(3), 277 - 86
Site-specific and reversible anchoring of active proteins onto cellulose using a cellulosome-like complex; Eklund M et al.; Protein engineering strategies facilitating controlled and spontaneous assembly of macromolecular complexes are of great interest for the design of artificial multi-enzyme systems of pre-defined composition . Here we have combined affinity proteins from different sources to achieve specific and reversible anchoring of affinity domain-tagged reporter proteins to a cellulose-anchored fusion protein . The design principle mimics the architecture of macromolecular cellulosome complexes produced by some cellulolytic microbes . A fusion protein between a cellulose-binding module (CBM1Cel6A) of the Trichoderma reesei cellobiohydrolase Cel6A and a five-domain staphylococcal protein A (SPA) was constructed to serve as platform for docking of easily detectable reporter proteins onto cellulose surfaces . In turn, the reporter proteins were produced as fusions to two copies of a SPA-binding affinity protein (an affibody denoted Z(SPA-1)), selected from a phage display library constructed by combinatorial protein engineering . In a series of experiments, involving repeated washing and low pH elution, affinity-tagged Enhanced Green Fluorescent Protein (EGFP) and Fusarium solani pisi lipase cutinase reporter proteins were both found to be specifically directed from solution to the same region of a cellulose filter paper where SPA-CBM1Cel6A fusion protein had been previously applied . This showed that the SPA-CBM1Cel6A fusion protein had been stably anchored to the cellulose surface without loss of binding capacity and that the interaction between SPA and the Z(SPA-1) affibody domains was selective . The generality of this biospecificity-driven system for assembly applications is discussed.

J Mol Biol, 2004 Apr 23, 338(2), 383 - 400
Early events during folding of wild-type staphylococcal nuclease and a single-tryptophan variant studied by ultrarapid mixing; Maki K et al.; A continuous-flow mixing device with a dead time of 100 micros coupled with intrinsic tryptophan and 1-anilinonaphthalene-8-sulfonate (ANS) fluorescence was used to monitor structure formation during early stages of the folding of staphylococcal nuclease (SNase) . A variant with a unique tryptophan fluorophore in the N-terminal beta-barrel domain (Trp76 SNase) was obtained by replacing the single Trp140 in wild-type SNase with His in combination with Trp substitution of Phe76 . A common background of P47G, P117G and H124L mutations was chosen in order to stabilize the protein and prevent accumulation of cis proline isomers under native conditions . In contrast to WT(*) SNase, which shows no changes in tryptophan fluorescence prior to the rate-limiting folding step ( approximately 100 ms), the F76W/W140H variant shows additional changes (enhancement) during an early folding phase with a time constant of 75 micros . Both proteins exhibit a major increase in ANS fluorescence and identical rates for this early folding event . These findings are consistent with the rapid accumulation of an ensemble of states containing a loosely packed hydrophobic core involving primarily the beta-barrel domain while the specific interactions in the alpha-helical domain involving Trp140 are formed only during the final stages of folding . The fact that both variants exhibit the same number of kinetic phases with very similar rates confirms that the folding mechanism is not perturbed by the F76W/W140H mutations . However, the Trp at position 76 reports on the rapid formation of a hydrophobic cluster in the N-terminal beta-sheet region while the wild-type Trp140 is silent during this early stage of folding . Quantitative modeling of the (un)folding kinetics and thermodynamics of these two proteins versus urea concentration revealed that the F76W/W140H mutation selectively destabilizes the native state relative to WT(*) SNase while the stability of transient intermediates remains unchanged, leading to accumulation of intermediates under equilibrium conditions at moderate denaturant concentrations.

Biochemistry, 2004 Apr 13, 43(14), 4064 - 70
Direct demonstration of structural similarity between native and denatured eglin C; Ohnishi S et al.; To characterize the long-range structure that persists in the unfolded form of the 70-residue protein eglin C, residual dipolar couplings (RDCs) for HN-N and HA-CA bond vectors were measured by NMR spectroscopy for both its low pH, urea denatured state and its native state . When the data sets for the two different structural states were compared, a statistically significant correlation was found, with both sets of dipolar couplings yielding a correlation coefficient of r = 0.47 to 0.51 . This finding directly demonstrates that the denatured state of eglin C has a nativelike global structure, a conclusion reached indirectly for staphylococcal nuclease by combining two different types of NMR data . A simple computer simulation showed that the degree of variation in phi and psi angles that yields the RDC correlation of r = 0.5 was inversely dependent on the statistical segment length, ranging from +/-6 to +/-30 degrees at the upper limit . Stable nativelike topologies that persist on unfolding would explain the rapid refolding kinetics displayed by many proteins and might provide a natural barrier against amyloid fibril formation.

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