Pneumologie, 2000 Sep, 54(9), 392 - 4 {Spontaneous lysis of pulmonary aspergillosis: "Aspergillus destroyed by Actinomyces"}; Hartmann J et al.; A 79 years old patient developed a large pulmonary aspergilloma in the cavities of his right upper lobe after postinfarctional pneumonia with local abscess formation . The clinical follow-up was characterized by recurrent hemoptysis resulting in marked anemia as well as by a continuous growth of the mycetoma . Suddenly a purulent gangrene of the whole upper lobe occurred infected by actinomyces israelii and staphylococcus but not aspergillus as it could be demonstrated in specimens from repeated transthoracic needle aspirations . After percutaneously inserted chest-tube drainage during 30 days the upper lobe cavity cleared up and the previously impressive aspergilloma had disappeared completely, however, the serum precipitins from aspergillus fumigatus still remained positive . After a course of several months without further pulmonary complications the patient finally died from a ruptured aortic aneurysm . It is suggested, that the spontaneous lysis of the aspergilloma was due to a deprivation of it' nutritive basis by the infected pulmonary tissue . A similar mechanism may also account for a sometimes successful treatment of pulmonary aspergilloma after injection of an amphotericin containing paste as a novel therapeutic strategy which is recommended in the case of patient's inoperable conditions. Nephrol Dial Transplant, 2000 Nov, 15(11), 1827 - 34 Catalytically active iron and bacterial growth in serum of haemodialysis patients after i.v . iron-saccharate administration; Parkkinen J et al.; BACKGROUND: I.v . iron is commonly administered to haemodialysis patients suffering from anaemia to improve their response to erythropoietin therapy . It has been unclear whether routinely used doses of i.v . iron preparations could result in iron release into plasma in amounts exceeding the iron binding capacity of transferrin . Here, we have studied the effect of 100 mg of iron saccharate given as an i.v . injection on transferrin saturation and the appearance of potentially harmful catalytically active iron . METHODS: We followed serum iron, transferrin and transferrin-saturation before and 5-210 min after administration of iron saccharate in 12 patients on chronic haemodialysis due to end-stage renal disease . We measured catalytically active iron by the bleomycin-detectable iron (BDI) assay and transferrin iron forms by urea gel electrophoresis, and studied iron-dependent growth of Staphylococcus epidermidis inoculated into the serum samples in vitro . RESULTS: The iron saccharate injection resulted in full transferrin saturation and appearance of BDI in the serum in seven out of the 12 patients . BDI appeared more often in patients with a low serum transferrin concentration, but it was not possible to identify patients at risk based on serum transferrin or ferritin level before i.v . iron . The average transferrin saturation and BDI level increased until the end of the follow-up time of 3.5 h . The appearance of BDI resulted in loss of the ability of patient serum to resist the growth of S . epidermidis, which was restored by adding iron-free apotransferrin to the serum . Iron saccharate, added to serum in vitro, released only little iron and promoted only slow bacterial growth, but caused falsely high transferrin saturation by one routinely used serum iron assay . CONCLUSIONS: The results indicate that 100 mg of iron saccharate often leads to transferrin oversaturation and the presence of catalytically active iron within 3.5 h after i.v . injection . As catalytically active iron is potentially toxic and may promote bacterial growth, it may be recommendable to use dosage regimens for i.v . iron that would not cause transferrin oversaturation. Singapore Med J, 2000 Apr, 41(4), 177 - 8 Staphylococcus lugdunensis: report of first case of skin and soft tissue infection in Singapore; Ling ML et al.; We report the first case of skin and soft tissue infection due to Staphylococcus lugdunensis in Singapore . This is a coagulase negative Staphylococcus species known to cause a wide variety of more serious infections--brain abscess, sepsis, chronic osteomyelitis and infective endocarditis. J Antimicrob Chemother, 2000 Nov, 46(5), 751 - 6 Efficacy of polycationic peptides in preventing vascular graft infection due to Staphylococcus epidermidis; Giacometti A et al.; A rat model was used to investigate the efficacy of two polycationic peptides, ranalexin and buforin II, in the prevention of vascular prosthetic graft infection due to methicillin-resistant Staphylococcus epidermidis with intermediate resistance to glycopeptides . The in vitro activity of the peptides was compared with those of vancomycin and teicoplanin by MIC determination and time-kill study . Moreover, the efficacy of collagen-sealed peptide-soaked Dacron was evaluated in a rat model of graft infection . Graft infections were established in the dorsal subcutaneous tissue of 120 adult male Wistar rats . The in vivo study included a control group, one contaminated group that did not receive any antibiotic prophylaxis and four contaminated groups that received an antibiotic-soaked graft . Experiments demonstrated that the activities of buforin II and ranalexin were greater than those of vancomycin and teicoplanin . Particularly, rats with buforin II-coated Dacron grafts showed no evidence of staphylococcal infection while, for the rats with ranalexin-, vancomycin- and teicoplanin-coated Dacron grafts, the quantitative graft cultures demonstrated bacterial growth (1.9 x 10(2) +/- 0.6 x 10(2) cfu/mL, 6 . 2 x 103 +/- 1.9 x 10(3) cfu/mL and 5.1 x 10(4) +/- 4.8 x 10(3) cfu/mL, respectively) . The study demonstrated that the use of peptide-soaked Dacron graft can result in significant bacterial growth inhibition and indicates that these compounds may be potentially useful in prosthetic surgery. J Biochem (Tokyo), 2000 Nov, 128(5), 739 - 44 Double point mutant F34W/W140F of staphylococcal nuclease is in a molten globule state but highly competent to fold into a functional conformation; Li Y et al.; The double point mutant F34W/W140F of staphylococcal nuclease was created and then characterized by far and near-UV CD, size-exclusion chromatography, ANS-binding fluorescence . The results show that this mutant has properties consistent with the classical definition of a molten globule, i.e., substantial secondary structure but no unique tight packing of tertiary structure, a relatively compact size and a larger exposed hydrophobic surface area as compared with the wild type enzyme, indicating that a molten globule can occur under physiological conditions . However, the activity assay showed that the mutant still maintains wild-type levels of activity . To further clarify the mechanism of the substrate-induced reactivation, enzymatic parameters such as K(M)(DNA), K(S)(DNA), K(M)(Ca), K(A)(Ca), K(d)(pdTp), and V(max) were determined, showing that all the parameters of this mutant are similar to those of the wild type enzyme . The results indicate that the F34W/W140F mutant has a similar substrate affinity to the wild type enzyme, and the functional conformation can be restored by substrate binding, which corresponds to the conformational adjustment capability of the mutant upon binding to ligands pdTp and Ca(2+) . The severely disrupted tertiary structure and high activity of the mutant indicate that it is highly competent to fold to its functional conformation . The results suggest that the primary structure can only guide the mutant to a molten globule state and that ligand-binding causes the mutant to fold further into its functionally active conformation, indicating that ligand-binding plays an important role in protein folding and catalysis. Appl Environ Microbiol, 2000 Nov, 66(11), 5083 - 6 Low sensitivity of Listeria monocytogenes to quaternary ammonium compounds; Mereghetti L et al.; Ninety-seven epidemiologically unrelated strains of Listeria monocytogenes were investigated for their sensitivities to quaternary ammonium compounds (benzalkonium chloride and cetrimide) . The MICs for seven serogroup 1/2 strains were high . Three came from the environment and four came from food; none were isolated from human or animal samples . All 97 strains carried the mdrL gene, which encodes a multidrug efflux pump, and the orfA gene, a putative transcriptional repressor of mdrL . The absence of plasmids in four of the seven resistant strains and the conservation of resistance after plasmid curing suggested that the resistance genes are not plasmid borne . Moreover, PCR amplification and Southern blot hybridization experiments failed to find genes phylogenetically related to the qacA and smr genes, encoding multidrug efflux systems previously described for the genus Staphylococcus . The high association between nontypeability by phages and the loss of sensitivity to quaternary ammonium compounds are suggestive of an intrinsic resistance due to modifications in the cell wall. Dig Surg, 2000, 17(4), 354 - 7 Splenic abscess . An old disease with new interest; Smyrniotis V et al.; BACKGROUND/AIMS: To study the demographics, signs and symptoms, causes, risk factors, imaging findings, bacteriologic profile, treatment and outcome of patients with splenic abscess . METHOD: The medical records of 17 patients with splenic abscess at two tertiary-care hospitals between 1989 and 1997 were retrospectively reviewed . The demographic data, physical and radiological findings, treatment, bacteriology reports and outcome of treatment were reviewed . RESULTS: The mean age of patients was 43 years (range 7-79 years) . Fever and abdominal pain were the most prominent signs . Seven patients were immunocompromised, three had abscessed hydatic cysts, two were drug users and three suffered from splenic trauma, infarction, and endocarditis, respectively . No predisposing factor was identified in 2 patients . In all cases, CT demonstrated the splenic lesion(s) . Staphylococcus species and Bacteriodes were the most common microbes, identified in the blood and abscess cultures . Thirteen patients underwent splenectomy, two medical therapy and two no therapy with respective survival rates of 92, 100 and 0% . CONCLUSION: Splenic abscess is a rare surgical entity encountered mostly in immunocompromised patients . CT scan is the gold standard for the definite diagnosis . Splenectomy is the treatment of choice, while medical therapy should be reserved for unusual pathogens provided that an effective antimicrobial agent is available . Vet Immunol Immunopathol, 2000 Oct 31, 76(3-4), 231 - 8 Presence of glutamine at position 74 of pocket 4 in the BoLA-DR antigen binding groove is associated with occurrence of clinical mastitis caused by Staphylococcus species; Sharif S et al.; Potential relationships between amino acid motifs in the antigen binding groove of various alleles of the bovine major histocompatibility complex DR (BoLA-DR) molecule and occurrence of clinical mastitis caused by Staphylococcus species (non-Staphylococcus aureus) were investigated in a case-control study . A significant association (P< or =0.05) was detected between the presence of glutamic acid at position beta 74 and occurrence of mastitis caused by Staphylococcus spp . with a relative risk of 11 . This motif is present in BoLA-DRB3.2*22, *23 and *24 alleles . Presence of a positively charged residue (arginine or lysine) at position 13 also showed a tendency (P< o r=0.1) towards an association with a higher risk of clinical mastitis caused by the same bacteria . This motif is present in BoLA-DRB3.2*23 and *8 alleles . Similarly, presence of arginine at position beta 71 (present in alleles *23 and *22) was associated with occurrence of this disease . These positions (beta 13, beta 71 and beta 74) form pocket 4 of the antigen binding groove, which plays an instrumental role in antigen binding and recognition by T-lymphocytes . Thus, it can be concluded that pocket 4 of the BoLA-DR molecule is involved in conferring susceptibility to clinical mastitis caused by Staphylococcus spp. Mol Cell Probes, 2000 Oct, 14(5), 311 - 9 Comparative molecular analysis of erythromycin-resistance determinants in staphylococcal isolates of poultry and human origin; Nawaz MS et al.; The ermA, ermB, ermC and msrA/msrB genes were detected in multidrug-resistant Staphylococcus spp . strains by PCR . Among 25 human clinical staphylococcal isolates the ermA, ermB, ermC and the msrA/msrB genes were detected in 88, 72, 4 and 100% of the strains, respectively . Among 24 poultry isolates the ermA, ermB, ermC and the msrA/msrB genes were detected in 100, 16.6, 50 and 12.5% of the strains, respectively . The ermA gene was found exclusively on the chromosome, whereas the ermC gene was found on 2.4-4.2 kb plasmids . Restriction fragment length polymorphism (RFLP) analysis of the ermA gene with Eco RI revealed five patterns (25.0, 21.0, 10.5, 6.2 and 4 . 8 kb) for the clinical strains and two (8.0 and 6.2 kb) for the poultry strains . The 6.2 kb RFLP pattern, in both the poultry and human clinical isolates, indicates a common lineage for the ermA gene . Antimicrob Agents Chemother, 2000 Nov, 44(11), 3122 - 6 Glycopeptide susceptibility profiles of Staphylococcus haemolyticus bloodstream isolates; Biavasco F et al.; Twelve clinical strains of Staphylococcus haemolyticus (eight methicillin resistant and three methicillin susceptible), isolated from blood cultures between 1982 and 1997, were investigated for teicoplanin and vancomycin susceptibility profiles . On the basis of conventional MIC tests and breakpoints, four isolates were susceptible (MICs, 1 to 8 microgram/ml) and eight were resistant (MICs, 32 to 64 microgram/ml) to teicoplanin while all were susceptible to vancomycin (MICs, 1 to 2 microgram/ml) . All four strains for which the conventional teicoplanin MICs were within the range of susceptibility expressed heterogeneous resistance to teicoplanin and homogeneous vancomycin susceptibility . Of the eight strains for which the conventional teicoplanin MICs were within the range of resistance, six expressed heterogeneous and two expressed homogeneous teicoplanin resistance while seven showed heterogeneous vancomycin resistance profiles (with subpopulations growing on 8 microgram of the drug per ml at frequencies of >/=10(-6) for six strains and 10(-7) for one) and one demonstrated homogeneous vancomycin susceptibility . Of six bloodstream isolates of other staphylococcal species (S . aureus, S . epidermidis, and S . simulans), for all of which the conventional teicoplanin MICs were >/=4 microgram/ml and the vancomycin MICs were </=2 microgram/ml, none exhibited heterogeneous susceptibility profiles for teicoplanin while three showed homogeneous and three showed heterogeneous susceptibility profiles for vancomycin (with subpopulations growing on 8 microgram of the drug per ml found for only one strain) . The results of this study indicate that a heterogeneous response to glycopeptides is a common feature of S . haemolyticus isolates and suggest that susceptibility to glycopeptides as determined by conventional MIC tests may not be predictive of the outcome of glycopeptide therapy. Infect Immun, 2000 Nov, 68(11), 6398 - 401 A novel polymorphism in the toll-like receptor 2 gene and its potential association with staphylococcal infection; Lorenz E et al.; The toll-like receptor 2 (TLR2) has gained importance as a major mammalian receptor for lipoproteins derived from the cell wall of a variety of bacteria, such as Borrelia burgdorferi, Treponema pallidum, and Mycoplasma fermentans . We were interested in identifying mutations in the TLR2 gene that might prove to be associated with altered susceptibility to septic shock . We performed a mutation screen of the TLR2 gene using single-stranded conformational polymorphism in 110 normal, healthy study subjects and detected an Arg753Gln mutation in three individuals . No other missense mutations were detected in the TLR2 open reading frame . Functional studies demonstrate that the Arg753Gln polymorphism, in comparison to the wild-type TLR2 gene, is significantly less responsive to bacterial peptides derived from B . burgdorferi and T . pallidum . In a septic shock population, the Arg753Gln TLR2 polymorphism occurred in 2 out of 91 septic patients . More importantly, both of the subjects with the TLR2 Arg753Gln polymorphism had staphylococcal infections . These findings suggest that a mutation in the TLR2 gene may predispose individuals to life-threatening bacterial infections. Diagn Microbiol Infect Dis, 2000 Oct, 38(2), 69 - 77 Structural analysis of a defective transfer-like region in a staphylococcal aminoglycoside resistance plasmid; von David W et al.; Staphylococcus epidermidis clinical isolate MH6502 contained the 51 . 9-kb nonconjugal plasmid pMH6502, which has homology to a major part of the transfer gene region of a known conjugal plasmid . Plasmid pMH6502 mediates aminoglycoside and ethidium bromide resistance . During restriction digest analysis of pMH6502, a double logarithmic regression of marker data gave a better linear relationship than a semi-logarithmic one . The analysis indicated several differences in the transfer-like region of pMH6502 compared to the analogous region of the S . aureus conjugal plasmid pG01 . The transfer-like region was in the opposite orientation compared to pG01 . An EcoRI site that is within the transfer-like region of pMH6502, has no analogue in pG01 . A HindIII site, located outside a 6.3-kb EcoRI fragment in the transfer gene region of pG01, is inside the analogous fragment of pMH6502 . A model is proposed to describe how a conjugal ancestral plasmid of pMH6502 could alter to its present form. J Immunol, 2000 Oct 1, 165(7), 3820 - 9 CD28 utilizes Vav-1 to enhance TCR-proximal signaling and NF-AT activation; Michel F et al.; The mechanism through which CD28 costimulation potentiates TCR-driven gene expression is still not clearly defined . Vav-1, an exchange factor for Rho GTPases thought to regulate, mainly through Rac-1, various signaling components leading to cytokine gene expression, is tyrosine phosphorylated upon CD28 engagement . Here, we provide evidence for a key role of Vav-1 in CD28-mediated signaling . Overexpression of Vav-1 in Jurkat cells in combination with CD28 ligation strongly reduced the concentration of staphylococcus enterotoxin E/MHC required for TCR-induced NF-AT activation . Surprisingly, upon Vav-1 overexpression CD28 ligation sufficed to activate NF-AT in the absence of TCR engagement . This effect was not mediated by overexpression of ZAP-70 nor of SLP-76 but necessitated the intracellular tail of CD28, the intactness of the TCR-proximal signaling cascade, the Src-homology domain 2 (SH2) domain of Vav-1, and SLP-76 phosphorylation, an event which was favored by Vav-1 itself . Cells overexpressing Vav-1 formed lamellipodia and microspikes reminiscent of Rac-1 and Cdc42 activation, respectively, for which the SH2 domain of Vav-1 was dispensable . Together, these data suggest that CD28 engagement activates Vav-1 to boost TCR signals through a synergistic cooperation between Vav-1 and SLP-76 and probably via cortical actin changes to facilitate the organization of a signaling zone. J Endovasc Ther, 2000 Oct, 7(5), 404 - 9 Suprarenal mycotic aneurysm exclusion using a stent with a partial autologous covering; Madhavan P et al.; PURPOSE: To report a combined endovascular and open technique to manage a suprarenal mycotic aortic aneurysm using a stent-graft partially covered with a section of autologous artery . METHODS AND RESULTS: A 50-year-old was hospitalized for staphylococcal septicemia and severe back pain . A previously diagnosed 3-cm abdominal aortic aneurysm was found to have expanded 2 cm in 3 weeks . Aortography documented some periaortic thickening and 2 mycotic aneurysms, one posterior at the level of the superior mesenteric artery and the second at the aortic bifurcation . After intensive antibiotic therapy, an endovascular approach to exclude the suprarenal mycotic aneurysm was undertaken in tandem with surgical excision of the infrarenal aneurysm . The harvested right common iliac artery was used to partially cover a Palmaz stent, which was deployed under direct vision just above the renal artery ostia so that the covered portion of the stent excluded the aneurysm . A right axillofemoral bypass with a femorofemoral bypass completed the revascularization . Postoperatively, the patient developed renal failure, ischemic colitis necessitating a left hemicolectomy, and paraplegia . Although the patient is paralyzed, the aneurysm remains excluded with patent visceral vessels at 12 months following surgery . No organisms were grown from excised aortic tissue, and no signs of recurrent infection have been seen . CONCLUSIONS: Stent-graft repair may be able to lessen the invasiveness and reduce the morbidity associated with treatment of mycotic aortic aneurysms. Arch Otolaryngol Head Neck Surg, 2000 Oct, 126(10), 1225 - 31 Systemic adoptive T-cell immunotherapy in recurrent and metastatic carcinoma of the head and neck: a phase 1 study; To WC et al.; OBJECTIVE: To evaluate the feasibility and toxic effects of systemic adoptive T-cell immunotherapy in patients with unresectable squamous cell carcinoma of the head and neck (SCCHN) . DESIGN: Nonrandomized phase 1 clinical trial . SETTING: Academic tertiary care hospital . PATIENTS: Between April 1, 1996, and September 30, 1998, 17 patients with confirmed recurrent and metastatic SCC of the upper aerodigestive tract were enrolled . Two patients did not receive T cells because of poor vaccine response . Fifteen patients were successfully treated with T-cell immunotherapy . INTERVENTION: Patients were vaccinated on the thigh with irradiated autologous tumor cells admixed with granulocyte-macrophage colony-stimulating factor (GM-CSF) followed by 3 additional daily injections of GM-CSF at the vaccination site . Eight to 10 days later, tumor cell vaccine-draining inguinal lymph nodes were resected, and lymph node lymphocytes were activated with staphylococcal enterotoxin A and expanded in interleukin 2 in vitro . Resulting cultured cells were infused into patients peripherally on an outpatient basis . RESULTS: Toxic effects of infusion were limited to grade 2 reactions in 3 of 16 treatments . One patient required overnight hospitalization for fever and emesis . Median cell expansion was 37 times (range, 4-416 times), and median cell dose was 7.5 x 10(9) (range, 1.3 x 10(8) to 4.2 x 10(10)) . Infused cells were predominantly CD3+ (>97%), being a mixture of CD4+ and CD8+ cells . Three patients demonstrated stabilization of previously progressive disease . Two patients experienced favorable clinical courses after adoptive T-cell transfer, including 1 patient with no evidence of disease 4 years after surgical resection of a vertebral body metastasis . CONCLUSIONS: Adoptive immunotherapy is a technically feasible and safe treatment with low toxicity and may demonstrate therapeutic activity in patients with unresectable SCCHN. Appl Microbiol Biotechnol, 2000 Sep, 54(3), 361 - 9 Fed-batch production of recombinant human calcitonin precursor fusion protein using Staphylococcus carnosus as an expression-secretion system; Dilsen S et al.; A pH-auxostatic fed-batch process was developed for the secretory production of a fusion protein consisting of the pro-part of Staphylococcus hyicus lipase and two synthetic human calcitonin (hCT) precursor repeats under the control of a xylose-inducible promotor from Staphylococcus xylosus . Using glycerol as the energy source and pH-controlled addition of yeast extract resulted in the production of 2000 mg 1(-1) of the fusion protein (420 mg 1(-1) of the recombinant hCT precursor) within 14 h, reaching 45 g 1(-1) cell dry mass with Staphylococcus carnosus in a stirred-tank reactor . Product titer and space-time yield (30 mg calcitonin precursor 1(-1) h(-1)) were thus improved by a factor of 2, and 4.5, respectively, compared to Escherichia coli expression-secretion systems for the production of calcitonin precursors . Two hundred grams of the fusion protein was secreted by the recombinant S . carnosus on a 150-1 scale (scale-up factor of 50) with a minimum use of technical-grade yeast extract (40 mg fusion protein g(-1) yeast extract). J Med Liban, 2000 Mar-Apr, 48(2), 77 - 83 Pattern of proven bacterial sepsis in a neonatal intensive care unit in Riyadh-Saudi Arabia: a 2-year analysis; Kilani RA et al.; Bacterial infections are an important cause of neonatal mortality and morbidity . The major pathogens for neonatal sepsis in the neonatal intensive care unit (NICU) vary with geographical area and time . It is therefore important to frequently audit neonatal sepsis in individual NICU, to aid in provision of adequate and appropriate preventive and therapeutic measures . We retrospectively reviewed the medical records of all infants who had positive blood cultures during a 2-year period in the NICU at a university hospital in Riyadh, Saudi Arabia . Overall the incidence of proven-bacterial-sepsis (PBS) was 10.2% of NICU admissions . The incidence of PBS in low-birth-weight (LBW), very low-birth-weight (VLBW), and extremely low-birth-weight (ELBW) infants were 19%, 41%, and 49% respectively . Multiple episodes of bacterial sepsis occurred in 21% of all infants infected . Coagulase negative Staphylococcus (CONS) (50%) was the most common infecting organism causing late onset sepsis (LOS) and Escherichia coli (29%) the most common causing early onset sepsis (EOS) . Gram negative bacteria (GNB) were the infecting organisms in 50% of the EOS episodes and 29% of LOS episodes . Only 11% (14) of the PBS were EOS . Only 10 (10.4%) infants had bacterial meningitis . The overall PBS related mortality was 9%, representing 22% of all neonatal deaths. Proc Natl Acad Sci U S A, 2000 Oct 10, 97(21), 11614 - 9 The osmolyte xylitol reduces the salt concentration of airway surface liquid and may enhance bacterial killing; Zabner J et al.; The thin layer of airway surface liquid (ASL) contains antimicrobial substances that kill the small numbers of bacteria that are constantly being deposited in the lungs . An increase in ASL salt concentration inhibits the activity of airway antimicrobial factors and may partially explain the pathogenesis of cystic fibrosis (CF) . We tested the hypothesis that an osmolyte with a low transepithelial permeability may lower the ASL salt concentration, thereby enhancing innate immunity . We found that the five-carbon sugar xylitol has a low transepithelial permeability, is poorly metabolized by several bacteria, and can lower the ASL salt concentration in both CF and non-CF airway epithelia in vitro . Furthermore, in a double-blind, randomized, crossover study, xylitol sprayed for 4 days into each nostril of normal volunteers significantly decreased the number of nasal coagulase-negative Staphylococcus compared with saline control . Xylitol may be of value in decreasing ASL salt concentration and enhancing the innate antimicrobial defense at the airway surface. Biophys J, 2000 Oct, 79(4), 1967 - 75 Interaction of the noncovalent molecular adapter, beta-cyclodextrin, with the staphylococcal alpha-hemolysin pore; Gu LQ et al.; Cyclodextrins act as noncovalent molecular adapters when lodged in the lumen of the alpha-hemolysin (alphaHL) pore . The adapters act as binding sites for channel blockers, thereby offering a basis for the detection of a variety of organic molecules with alphaHL as a biosensor element . To further such studies, it is important to find conditions under which the dwell time of cyclodextrins in the lumen of the pore is extended . Here, we use single-channel recording to explore the pH- and voltage-dependence of the interaction of beta-cyclodextrin (betaCD) with alphaHL . betaCD can access its binding site only from the trans entrance of pores inserted from the cis side of a bilayer . Analysis of the binding kinetics shows that there is a single binding site for betaCD, with an apparent equilibrium dissociation constant that varies by >100-fold under the conditions explored . The dissociation rate constant for the neutral betaCD molecule varies with pH and voltage, a result that is incompatible with two states of the alphaHL pore, one of high and the other of low affinity . Rather, the data suggest that the actual equilibrium dissociation constant for the alphaHL . betaCD complex varies continuously with the transmembrane potential. Thromb Haemost, 2000 Sep, 84(3), 374 - 80 Heterogeneous recognition of beta 2-glycoprotein I by antibodies from antiphospholipid syndrome patients; Guerin J et al.; Beta 2-glycoprotein I plays a pivotal role in the binding of antiphospholipid antibodies to phospholipid in patients with antiphospholipid syndrome . In this study the nature of the epitopes on beta 2-glycoprotein I (beta2-GPI) recognised by sera from antiphospholipid syndrome (APS) patients (n = 15) was investigated and compared to rabbit polyclonal and mouse monoclonal anti-beta2-GPI antibodies . beta2-GPI was only recognised when bound to a high affinity binding support . The antigenic epitope on beta2-GPI recognised by all APS patients was also dependent on disulphide bond integrity . Digestion of beta2-GPI with elastase rapidly destroyed the epitope(s) on beta2-GPI recognised by antibodies in 91% of APS patients . The main cleavage occurred at tryptophan316-lysine317 in the fifth domain . Digestion with staphylococcal V8 protease resulted in a 50% reduction in antibody binding in 81% of patients and the cleavage sites mainly involved the first domain of the molecule . There was considerable variability in the recognition of six different species of beta2-GPI by serum from APS patients . The epitopes on beta2-GPI bound by APS sera appear conformationally determined in all patients but are quite heterogeneous in the regions of beta2-GPI that are recognised. Nat Biotechnol, 2000 Oct, 18(10), 1091 - 5 Detecting protein analytes that modulate transmembrane movement of a polymer chain within a single protein pore; Movileanu L et al.; Here we describe a new type of biosensor element for detecting proteins in solution at nanomolar concentrations . We tethered a 3.4 kDa polyethylene glycol chain at a defined site within the lumen of the transmembrane protein pore formed by staphylococcal alpha-hemolysin . The free end of the polymer was covalently attached to a biotin molecule . On incorporation of the modified pore into a lipid bilayer, the biotinyl group moves from one side of the membrane to the other, and is detected by reversible capture with a mutant streptavidin . The capture events are observed as changes in ionic current passing through single pores in planar bilayers . Accordingly, the modified pore allows detection of a protein analyte at the single-molecule level, facilitating both quantification and identification through a distinctive current signature . The approach has higher time resolution compared with other kinetic measurements, such as those obtained by surface plasmon resonance. J Clin Microbiol, 2000 Oct, 38(10), 3887 - 9 Osteomyelitis caused by Staphylococcus schleiferi and evidence of misidentification of this Staphylococcus species by an automated bacterial identification system; Calvo J et al.; We report a case of sternal osteomyelitis due to Staphylococcus schleiferi in a patient who underwent thoracic surgery . This constitutes the first documented case of osteomyelitis caused by this Staphylococcus species . We also relate our experience in the utilization of commercially available MicroScan panels for the identification of this microorganism. Int J Food Microbiol, 2000 Sep 15, 60(1), 1 - 13 Multitoxin biosensor-mass spectrometry analysis: a new approach for rapid, real-time, sensitive analysis of staphylococcal toxins in food; Nedelkov D et al.; Biomolecular interaction analysis mass spectrometry (BIA-MS) was applied to detection of bacterial toxins in food samples . This two-step approach utilizes surface plasmon resonance (SPR) to detect the binding of the toxin(s) to antibodies immobilized on a surface of a sensor chip . SPR detection is then followed by identification of the bound toxin(s) by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry . Staphylococcal enterotoxin B (SEB) was readily detected in milk and mushroom samples at levels of 1 ng/ml . In addition, non-specific binding of food components to the immobilized antibody and to the sensor chip surface was detected . To evaluate the applicability of BIA-MS in the analysis of materials containing multiple toxic components, sample containing both SEB and toxic-shock syndrome toxin-1 was analyzed . Both toxins were successfully and simultaneously detected through the utilization of multiaffinity sensor chip surfaces. Gig Sanit, 1998 Nov-Dec, (6), 13 - 6 {Hygienic evaluation of staphylococcal nasal mucosal biocenosis in school children of an industrial city}; Usviatsov BIa et al.; The results of microbiological spatial monitoring in Orenburg were evaluated . A biological variety of Staphylococcus on the nasal mucosa of schoolchildren were detected . The highest proportion of carriage of the pathogen Staphylococcus was found in different districts of Orenburg, which had the highest air pollution. Gig Sanit, 1998 Nov-Dec, (6), 8 - 11 {Microbiological analysis of the state of environment in the Orenburg region}; Bukharin OV et al.; The frequency of the residual staphylococcal carriage among the children who live in rural area having different levels and patterns of environmental pollution is estimated . A relationship is found between the frequency of the residual staphylococcal carriage and the rate of technogenic pollution of the ambient air . The findings suggest that the staphylococcal carriage among children may be used as an index of the unfavorable environment. Avian Dis, 2000 Jul-Sep, 44(3), 737 - 44 Isolation of avian influenza virus (H10N7) from an emu (Dromaius novaehollandiae) with conjunctivitis and respiratory disease; Woolcock PR et al.; Avian influenza virus was isolated from the conjunctiva of a male emu chick . Clinical observations included ocular discharge, dyspnea, and mild respiratory signs . Lesions included conjunctivitis, tracheitis, bronchopneumonia, and airsacculitis . Escherichia coli was isolated from the conjunctiva and the sinus, and Staphylococcus sp . was isolated from the conjunctiva . Influenza A viral nucleoprotein was detected immunohistochemically in epithelial cells of the bronchi, lung parenchyma and tracheal mucosa, and mononuclear inflammatory cells within the exudate of the bronchial lumen; conjunctiva, air sacs, kidney, intestine, and liver were negative for the viral nucleoprotein . The isolated influenza virus was typed as H10N7 and was determined to be nonpathogenic for chickens. Cell Immunol, 2000 Aug 25, 204(1), 1 - 10 Effect of Staphylococcus enterotoxin B on the concurrent CD8(+) T cell response to influenza virus infection; Huang CC et al.; Bacterial superantigens have potent in vivo effects . Respiratory viral infections are often associated with secondary bacterial infections, raising the likelihood of exposure to bacterial superantigens after the initiation of the anti-viral immune response . In this study, the general and V beta-specific effects of exposure to Staphylococcal enterotoxin B (SEB) during influenza virus infection on both the ongoing acute and the subsequent recall CD8(+) T cell responses were analyzed, using the well-characterized murine influenza model system and tetrameric MHC/peptide reagents to directly identify virus-specific T cells . The results show that although superantigen exposure during the primary viral infection caused delayed viral clearance, there was remarkably little effect of SEB on the magnitude or TCR repertoire of the ongoing cytolytic T cell response or on the recall response elicited by secondary viral infection . Thus, despite the well-characterized immunomodulatory effects of SEB, there was surprisingly little interference with concurrent anti-viral immunity . An Esp Pediatr, 2000 Mar, 52(3), 263 - 6 {Toxic shock syndrome in children . Report of four cases}; de Vincente Aymat A et al.; We report four children with toxic shock syndrome admitted in the pediatric intensive care unit of our hospital during the past year . All the children had the five criteria established by the Centers for Disease Control for the diagnosis of this syndrome . In all four there was a probable point of entry of the infection: maxillar sinusitis in one, pneumonia in two and surgical wound in the other . No bacteria that could have caused the infection were isolated in any of the children, which suggests a staphylococcal origin for this syndrome . Evolution was good in all of the children due to aggressive treatment that included inotropic support, volemic expansion and antibiotics . Two of the children, who suffered adult respiratory distress syndrome, required prolonged respiratory support. J Perinatol, 2000 Sep, 20(6), 384 - 6 Solitary hepatic abscess with associated glomerulonephritis in a neonate; DeFranco PE et al.; A full-term neonate with a history of umbilical venous catheterization followed by coagulase-negative staphylococcal sepsis is presented . The infant developed a solitary hepatic abscess with saprophytic organisms . Her liver abscess resulted in acute glomerulonephritis characterized by hypertension, proteinuria, oliguria, and azotemia . Surgical drainage and antibiotic treatment of the abscess was associated with resolution of the glomerulonephritis . Glomerulonephritis due to solitary liver abscess in a neonate has not been reported previously . Acute onset of glomerulonephritis should prompt a search for occult sources of infection. Arch Dis Child, 2000 Oct, 83(4), 353 - 5 Recurrent skin peeling following Kawasaki disease; Michie C et al.; Long term follow up of 259 cases of Kawasaki disease led to the observation that 11% of children have episodes of recurrent peeling of the skin for several years after their recovery . These events were usually associated with an upper respiratory tract infection and were distinct from a recurrence of Kawasaki disease . Repeeling was significantly less frequent in children who had suffered coronary artery dilatation and was more frequently seen in those with nasal staphylococcal colonisation . The mechanism for this phenomenon is unclear, but it has been observed in a number of other conditions caused by infectious agents and their toxins . Paediatricians need to be aware of this phenomenon which is distinct from recurrence of Kawasaki disease. J Invest Dermatol, 2000 Oct, 115(4), 746 - 52 Glucocorticoids augment the chemically induced production and gene expression of interleukin-1alpha through NF-kappaB and AP-1 activation in murine epidermal cells; Miyazaki Y et al.; To clarify the mechanism of the glucocorticoid-induced augmentation of skin response, we attempted to demonstrate the modulatory effect of glucocorticoids on the regulation of cytokines produced by keratinocytes stimulated with various chemicals in vitro . Haptens, irritants, and a superantigen (staphylococcal enterotoxin B) induced a significant release of interleukin-1alpha and tumor necrosis factor alpha, but not interleukin-10, from a murine keratinocyte cell line, Pam 212 cells . Glucocorticoids (10(-6)-10(-12) M) significantly augmented the production of interleukin-1alpha by Pam 212 cells at both the protein and mRNA levels when stimulated by either haptens or irritants, but not by staphylococcal enterotoxin B, whereas glucocorticoids alone had no effect . In contrast, glucocorticoids had no effect on the production of tumor necrosis factor alpha and interleukin-10 by chemically stimulated Pam 212 cells . Electrophoretic mobility shift assays revealed that chemical stimulation induced NF-kappaB activation in Pam 212 cells; however, augmented NF-kappaB activation by 10(-6)-10(-8) M of glucocorticoids was observed in Pam 212 cells stimulated by both haptens and irritants, but not by staphylococcal enterotoxin B . Furthermore, pyrrolidine dithiocarbamate inhibited the hapten-induced interleukin-1alpha production and NF-kappaB expression by Pam 212 cells . Pyrrolidine dithiocarbamate did not completely abrogate the hapten-induced interleukin-1alpha production augmented by glucocorticoids, however . To determine the effect on transcription factors other than NF-kappaB, AP-1 activity was examined by electrophoretic mobility shift assays . Hapten was founded to induce AP-1 activation in Pam 212 cells . In addition, AP-1 activation was augmented in the hapten-stimulated Pam 212 cells in the presence of 10(-8)-10(-10) M of glucocorticoids . The augmented inflammatory reaction by glucocorticoids may therefore reflect the augmentation of interleukin-1alpha production by keratinocytes mediated through the NF-kappaB and AP-1 pathway. Nahrung, 2000 Aug, 44(4), 272 - 5 Effect of preservation techniques and food additives on staphylococcal thermonuclease; Kumar JK et al.; Staphylococcal TNase was found to retain its activity fully even after exposure to chilling and refrigeration temperatures for 24 h . It was not inhibited by p-hydroxy benzoic acid, sorbic acid, methylpropyl p-benzoic acid and sodium nitrite in the concentration range of 0.04 to 0.5%, whereas it was inhibited by 100 ppm of butylated hydroxy anisole (BHA), 200 ppm of butylated hydroxy toluene (BHT), and 300 ppm of propyl gallate . There was not complete inhibition of S . aureus and TNase by tocopherol (TP) and ascorbic acid (AA) even at concentration of 300 ppm . These results indicate that TNase can be used as an index of potentially enterotoxin producing S . aureus contamination in foods subjected to chilling, refrigeration as well as in foods containing common preservatives and antioxidants. J Urol, 2000 Oct, 164(4), 1275 - 6 Is it safe to reuse a syringe of glutaraldehyde cross-linked collagen? A microbiological study; Culligan PJ et al.; PURPOSE: We evaluated the safety of saving partially used syringes of glutaraldehyde cross-linked collagen for subsequent treatment sessions in an individual . MATERIALS AND METHODS: After periurethral injection in an office setting 56 partially used syringes of glutaraldehyde cross-linked collagen were stored in a refrigerator for 1 to 61 weeks (mean 15) . Collagen from all 56 syringes was then cultured qualitatively using a broth medium at 35C and semiquantitatively using a chocolate agar plate at 22 to 30C for 5 days each . RESULTS: A qualitative broth culture was positive for coagulase negative staphylococcus but the results of semiquantitative chocolate agar culture of material from the same syringe were negative . All cultures of the other 55 syringes were negative . CONCLUSIONS: The positive culture most likely resulted from contamination during periurethral injection or the culturing process . Minimal contamination from and the great potential cost savings of reusing glutaraldehyde cross-linked collagen for subsequent treatments in an individual indicate the need for an expanded study involving multiple centers. Biotechnol Bioeng, 2000 Nov 5, 70(3), 349 - 52 Stereochemistry of nonnatural aldol reactions catalyzed by DHAP aldolases; Schoevaart R et al.; A coupled enzymatic assay was developed for quantitative determination of the stereoisomeric products formed in aldol reactions catalyzed by dihydroxyacetone phosphate (DHAP)-dependent aldolases . Three of the four stereoisomers could be determined directly; the fourth one was calculated . This procedure is based on the reversibility of the aldol reaction and requires no derivatization or work-up of the product samples, only removal or inactivation of the biocatalyst . In comparison with other methods the enzymatic assay is highly accurate and fast . Determination of isomer formation with 10 different acceptor substrates applying this procedure gave unprecedented insight in the stereochemistry of fructose-1,6-bisphosphate aldolase from Staphylococcus carnosus and l-rhamnulose-1-phosphate aldolase from E . coli . Antimicrob Agents Chemother, 2000 Oct, 44(10), 2842 - 4 Mupirocin prophylaxis against methicillin-susceptible, methicillin-resistant, or vancomycin-intermediate Staphylococcus epidermidis vascular-graft infection; Giacometti A et al.; A rat model was used to investigate the efficacy of mupirocin in the prevention of vascular prosthetic graft infection due to Staphylococcus epidermidis strains with different susceptibility patterns (methicillin susceptible, methicillin resistant, and with intermediate resistance to vancomycin) . The effect of mupirocin-soaked Dacron was compared to that of perioperative intraperitoneal prophylaxis with vancomycin . Graft infections were established in the back subcutaneous tissue of adult male Wistar rats by implantation of Dacron prostheses (1 cm(2)) followed by topical inoculation with 5 x 10(7) CFU of one staphylococcal strain . The study included a control group (no graft contamination), three contaminated groups that did not receive any antibiotic prophylaxis, three contaminated groups that received mupirocin-soaked grafts, three contaminated groups in which perioperative intraperitoneal vancomycin prophylaxis (10 mg/kg of body weight) was administered, and three contaminated groups that received mupirocin-soaked grafts and perioperative intraperitoneal vancomycin prophylaxis (10 mg/kg) . The grafts were sterilely removed 7 days after implantation, and the infection was evaluated by using sonication and quantitative agar culture . Data analysis showed the efficacy of mupirocin against all three strains, with growth of the strains in treated rats significantly different than that in the untreated control . In addition, mupirocin was more effective than vancomycin against the strain with intermediate susceptibility to the glycopeptide . Finally, the combination of mupirocin and vancomycin produced complete suppression of the growth of all of the strains. J Rheumatol, 2000 Sep, 27(9), 2152 - 9 Anti-beta2-glycoprotein I autoantibodies, in vitro thrombin generation, and the antiphospholipid syndrome; Hanly JG et al.; OBJECTIVE: Thrombin plays a pivotal role in the regulation of hemostasis . We examined the effect of antiphospholipid (aPL) antibodies, in particular those with specificity for beta2-glycoprotein I (beta2-GPI), on in vitro thrombin generation, and examined the association with clinical manifestations of the antiphospholipid syndrome (APS) . METHODS: We studied plasma samples from 59 patients with aPL antibodies determined by the presence of either elevated anticardiolipin (aCL) antibodies or lupus anticoagulant (LAC) . Direct antibody binding of IgG, IgM, and IgA to beta2-GPI and prothrombin (PT) was determined by ELISA . Affinity purification of total IgG and IgG anti-B2-GPI antibodies was performed using staphylococcal protein A and phospholipid liposomes . A chromogenic assay was used to determine the effect of plasma samples and purified autoantibodies on in vitro thrombin generation . RESULTS: Thirty-three of 59 (56%) plasma samples inhibited in vitro generation of thrombin and 7/59 (12%) accelerated thrombin formation . There was a strong negative correlation between thrombin generation and IgG aCL (r = -0.72, p < 0.001) and IgG anti-beta2-GPI (r = -0.71, p < 0.001) antibody levels, and a weaker correlation with LAC (r = -0.46, p = 0.001) . This association was not found with anti-PT antibodies and could not be attributed to concurrent therapy with warfarin . Additional experiments with affinity purified IgG antibodies indicated a dose dependent inhibition of thrombin generation, which was restricted to anti-beta2-GPI antibodies . Patients with a history of core clinical manifestations of the APS {venous and arterial thrombosis, recurrent (> or = 2) fetal loss} had significantly greater inhibition of in vitro thrombin generation (mean +/- SEM Z score: -3.38 +/- 0.51 vs -1.42 +/- 0.56; p = 0.01) and higher levels of IgG aCL (mean +/- SEM Z score: 8.39 +/- 1.12 vs 5.39 +/- 0.88; p = 0.04) and IgG anti-beta2-GPI antibodies (mean +/- SEM Z score: 4.49 +/- 0.69 vs 2.26 +/- 0.54; p = 0.01) . Odds ratios for these variables and clinical manifestations of the APS were 5.43, 4.17, and 3.28, respectively . CONCLUSION: aPL antibodies may accelerate or inhibit the rate of in vitro thrombin formation . The predominant effect is inhibition that is restricted to IgG anti-beta2-GPI antibodies and it is strongly associated with clinical manifestations of the APS. Structure Fold Des, 2000 Sep 15, 8(9), 961 - 9 The X-ray structure of the FMN-binding protein AtHal3 provides the structural basis for the activity of a regulatory subunit involved in signal transduction; Albert A et al.; BACKGROUND: The Arabidopsis thaliana HAL3 gene product encodes for an FMN-binding protein (AtHal3) that is related to plant growth and salt and osmotic tolerance . AtHal3 shows sequence homology to ScHal3, a regulatory subunit of the Saccharomyces cerevisae serine/threonine phosphatase PPz1 . It has been proposed that AtHal3 and ScHal3 have similar roles in cellular physiology, as Arabidopsis transgenic plants that overexpress AtHal3 and yeast cells that overexpress ScHal3 display similar phenotypes of improved salt tolerance . The enzymatic activity of AtHal3 has not been investigated . However, the AtHal3 sequence is homologous to that of EpiD, a flavoprotein from Staphylococcus epidermidis that recognizes a peptidic substrate and subsequently catalyzes the alpha, beta-dehydrogenation of its C-terminal cysteine residue . RESULTS: The X-ray structure of AtHal3 at 2 A resolution reveals that the biological unit is a trimer . Each protomer adopts an alpha/beta Rossmann fold consisting of a six-stranded parallel beta sheet flanked by two layers of alpha helices . The FMN-binding site of AtHal3 contains all the structural requirements of the flavoenzymes that catalyze dehydrogenation reactions . Comparison of the amino acid sequences of AtHal3, ScHal3 and EpiD reveals that a significant number of residues involved in trimer formation, the active site, and FMN binding are conserved . This observation suggests that ScHal3 and EpiD might also be trimers, having a similar structure and function to AtHal3 . CONCLUSIONS: Structural comparisons of AtHal3 with other FMN-binding proteins show that AtHal3 defines a new subgroup of this protein family that is involved in signal transduction . Analysis of the structure of AtHal3 indicates that this protein is designed to interact with another cellular component and to subsequently catalyze the alpha,beta-dehydrogenation of a peptidyl cysteine . Structural data from AtHal3, together with physiological and biochemical information from ScHal3 and EpiD, allow us to propose a model for the recognition and regulation of AtHal3/ScHal3 cellular partners. J Mol Biol, 2000 Sep 22, 302(3), 527 - 37 The crystal structure of staphylococcal enterotoxin H: implications for binding properties to MHC class II and TcR molecules; Hakansson M et al.; The X-ray structure of the superantigen staphylococcal enterotoxin H (SEH) has been determined at 1.69 A resolution . In this paper we present two structures of zinc-free SEH (apoSEH) and one zinc-loaded form of SEH (ZnSEH) . SEH exhibits the conventional superantigen (SAg) fold with two characteristic domains . In ZnSEH one zinc ion per SEH molecule is bound to the C-terminal beta-sheet in the region implicated for major histocompatibility complex class II (MHC class II) binding in SEA, SED and SEE . Surprisingly, the zinc ion has only two ligating amino acid residues His206 and Asp208 . The other ligands to the zinc ion are two water molecules . An extensive packing interaction between two symmetry-related molecules in the crystal, 834 A(2)/molecule, forms a cavity that buries the zinc ions of the molecules . This dimer-like interaction is found in two crystal forms . Nevertheless, zinc-dependent dimerisation is not observed in solution, as seen in the case of SED . A unique feature of SEH as compared to other staphylococcal enterotoxins is a large negatively charged surface close to the Zn(2+) site . The interaction of SEH with MHC class II is the strongest known among the staphylococcal enterotoxins . However, SEH seems to lack a SEB-like MHC class II binding site, since the side-chain properties of structurally equivalent amino acid residues in SEH and those in SEB-binding MHC class II differ dramatically . There is also a structural flexibility between the domains of SEH . The domains of two apoSEH structures are related by a 5 degrees rotation leading to at most 3 A difference in C(alpha) positions . Since the T-cell receptor probably interacts with both domains, SEH by this rotation may modulate its binding to different TcR Vbeta-chains . Immunity, 2000 Aug, 13(2), 179 - 86 Intrathymic T cell development and selection proceeds normally in the absence of glucocorticoid receptor signaling; Purton JF et al.; Glucocorticoids are believed to play a role in T cell development and selection, although their precise function is controversial . Glucocorticoid receptor (GR)-deficient mice were used to directly investigate this problem . GR-deficient thymocytes were resistant to dexamethasone-mediated apoptosis, confirming the absence of glucocorticoid responsiveness . An absence of GR signaling had no impact on thymocyte development either in vivo or in vitro . T cell differentiation, including positive selection, was normal as assessed by normal development of CD4+CD8+, alphabetaTCR+CD4+, and alphabetaTCR+CD8+ thymocytes . Negative selection, mediated by the superantigen staphylococcal enterotoxin B (SEB), or anti-CD3/CD28, was also normal in the absence of GR signaling . In contrast to earlier reports, these data demonstrate that GR signaling is not essential for intrathymic T cell development or selection. FEMS Microbiol Lett, 2000 Sep 1, 190(1), 115 - 20 Identification of beta-oxidation and thioesterase activities in Staphylococcus carnosus 833 strain; Engelvin G et al.; Staphylococcus carnosus 833, inoculated into sausage meat, increased the level of methyl ketones, which contributed to the cured aroma . These ketones can arise from incomplete beta-oxidation followed by two enzymatic activities: a thioesterase and a decarboxylase . In this study we identified the beta-oxidative pathway (through the measure of 3-hydroxyacyl-CoA dehydrogenase activity) and the thioesterase activity in extracts of S . carnosus cells grown in the presence of different methyl esters . The beta-oxidative system was induced by methyl esters and highest induction was found with a 12-carbon substrate . It was specific for medium chain length fatty acyl CoA substrates . Its maximal activity was observed at the end of stationary growth phase . HPLC analyses of acyl-CoA after incubation of cell extracts with palmitoyl-CoA showed that the beta-oxidation system released preferentially long chain hydroxyacyl-CoAs, enoyl-CoAs, and acyl-CoAs . The time-course of intermediate formation indicated a precursor product relationship indicative of a model of free intermediates which could be further deacylated by a thioesterase . The thioesterase activity was enhanced when S . carnosus was grown in the presence of methyl esters with at least 12 carbons and this enzyme was specific for short chain acyl-CoAs . The maximal activity was reached at the stationary growth phase. Isr Med Assoc J, 2000 Aug, 2(8), 598 - 600 Hepatitis A associated with other focal infections; Klar A et al.; BACKGROUND: Concomitant bacterial and viral infection is a well-known phenomenon, however only very rarely has a bacterial infection been reported during hepatitis A virus infection . OBJECTIVE: To evaluate retrospectively the clinical records of children hospitalized with HAV infection for a concomitant infection proved or presumed to be bacterial . METHOD: A retrospective study was conducted on all the children hospitalized with hepatitis A infection from 1988-96 in our center . The records were evaluated for a concomitant infection . RESULTS: Of 40 children hospitalized with HAV infection, 13 were found to have a concomitant infection: these included 6 with pneumonia, 4 with pyelonephritis and 1 case each of purulent otitis media, osteomyelitis and staphylococcal bacteremia . CONCLUSION: In areas where hepatitis A is endemic, a simultaneous infection with hepatitis A and other common bacterial infection during childhood may co-exist . A permissive role for HAV infection is suggested. Cytokine, 2000 Sep, 12(9), 1348 - 55 Interleukin 10 (IL-10)-mediated inhibition of inflammatory cytokine production by human alveolar macrophages; Raychaudhuri B et al.; Alveolar macrophages are an important source of inflammatory cytokines in the lung . IL-10 has been shown to inhibit inflammatory cytokine production by human alveolar macrophages, but mechanisms are unclear . The purpose of the present study was to investigate whether IL-10 modified cytokine production by interference with transcriptional pathways . Alveolar macrophages were obtained from healthy controls by fiberoptic bronchoscopy and incubated with LPS+/-IL-10 . Results indicated that steady state mRNA levels of tumour necrosis factor-alpha (TNF) and interleukin 1-beta (IL-1) decreased in the presence of IL-10 . Consequently, electrophoretic mobility shift assays were performed using end-labelled nuclear factor-kappa B (NF-kappa B) or activator protein-1 (AP-1) probe . NF-kappa B binding was decreased in extracts from macrophages incubated for 4 h with LPS+IL-10 in comparison to those incubated with LPS alone . IL-10 also inhibited TNF secretion and NF-kappa B activation induced by another stimulus, staphylococcal toxin . Supershift assays revealed the presence of both p50 and p65 subunits of NF-kappa B . AP-1 was not affected by IL-10 . Further examination of mechanisms indicated that IL-10 delayed the LPS-mediated degradation of the inhibitor protein I kappa B, thus delaying the nuclear translocation of the p65 subunit . These observations provide the first evidence that IL-10 antagonizes cytokine transcription in human alveolar macrophages by impeding the nuclear translocation of NF-kappa B by delaying the degradation of I kappa B . Ther Apher, 2000 Aug, 4(4), 275 - 9 Therapeutic apheresis in myasthenia gravis; Batocchi AP et al.; Plasma exchange (PE) is an easily applicable technique for rapid and massive removal of antibodies, and its beneficial role is well established in the management of myasthenia gravis (MG), an antibody-mediated disorder of the neuromuscular junction . PE is useful in myasthenic crisis, in most severe forms of MG before thymectomy, in the early postoperative period, and in cases of symptom worsening during tapering or initiation of immunosuppressive therapy . Clinical efficacy varies from 55% to 100%, and improvement rarely persists for more than 4-10 weeks; thus immunosuppressive therapy has to be associated . New apheretic techniques (double filtration plasmapheresis, immunoadsorption systems with staphylococcal protein A columns or thryptophan-polyvinyl alcohol gel columns) that allow the selective removal of IgG and anti-AChR antibody were recently used in the management of MG with positive effects . Whether their therapeutic effect and cost effect prove more favorable than those obtained by PE still must be demonstrated. Pediatr Nephrol, 2000 Sep, 14(10-11), 912 - 5 Rapid removal of vancomycin by continuous veno-venous hemofiltration; Shah M et al.; We describe a 14-year-old girl with staphylococcal (coagulase-negative) ventriculo-peritoneal shunt infection, who developed oliguric acute renal failure and was found to have a serum vancomycin concentration of 250 microg/ml . Since only about 10%-50% of vancomycin is bound to protein in blood, we employed continuous veno-venous hemofiltration (CVVH) with a high ultrafiltration rate (1,800 ml/h) for increased convective clearance to remove vancomycin, which may have contributed to the acute renal failure . At the end of 38 h of CVVH, the vancomycin concentration had decreased in an exponential manner to 27 microg/ml . Over the subsequent 3-4 days, her renal function improved and the vancomycin concentration decreased further to <5 microg/ml . In conclusion, we believe that a high serum vancomycin concentration may be nephrotoxic and demonstrate that CVVH can be used effectively to remove vancomycin in children with acute renal failure. Lett Appl Microbiol, 2000 Sep, 31(3), 228 - 32 Biogenic amine formation and oxidation by Staphylococcus xylosus strains from artisanal fermented sausages; Martuscelli M et al.; Fifty strains of Staph . xylosus, isolated from artisanal fermented sausages in Southern Italy (Lucania region) were tested to verify their potential to produce or degrade biogenic amines . Twenty-six strains analysed were not able to form amines, but seven had the potential to produce spermine and/or spermidine and, at lower levels, tryptamine and tyramine . By contrast, about 80% of the strains that did not possess amino acid decarboxylase activity, exhibited an ability to degrade histamine . The greatest histamine-oxidase activity was present in the strains S81 (100% degradation), S206 (93%), S79 (68%) and S90 (53%) . The strain S142 exhibited a remarkably high potential to oxidase tyramine and histamine, reducing the initial concentrations by 63 and 47%, respectively. Diabet Med, 2000 Jul, 17(7), 546 - 9 Blindness following a diabetic foot infection: a variant to the 'eye-foot syndrome'? Yuen KC, Baker NR, Reddy A, Edelsten C, Rayman G. AIMS: The 'eye-foot syndrome' was initially described by Walsh et al . to highlight the important association of foot lesions in patients with diabetic retinopathy . We present a case of a 58-year-old patient with Type 2 diabetes mellitus who developed blindness following endogenous staphylococcal endophthalmitis from an infected foot ulcer . RESULTS: Our case describes the link between the eye and the foot but is somewhat different to the association as described by Walsh et al . Endogenous endophthalmitis is rare with diabetic patients being especially at risk, and we report the first case of endogenous staphylococcal endophthalmitis related to a diabetic foot lesion . CONCLUSIONS: Our case illustrates several important issues in the management of diabetic patients admitted to hospital with infection; the need to thoroughly examine the feet to ascertain any foot lesions and any underlying peripheral vascular disease or peripheral neuropathy, to treat aggressively any infected foot lesions to prevent serious complications of septicaemia and to consider rare conditions like endogenous endophthalmitis in any diabetic patient presenting with acute visual impairment and septicaemia. Br J Haematol, 2000 Aug, 110(2), 461 - 8 Central venous access devices in children with congenital coagulation disorders: complications and long-term outcome; McMahon C et al.; Reliable venous access is essential to facilitate the administration of prophylactic factor concentrate or blood products in children with congenital coagulation disorders and immune tolerance therapy (ITT) regimens in those who develop high responding inhibitors . Poor venous access is even more problematic in very young children, the vast majority of whom will require the insertion of central venous access devices (CVADs) . Previous studies have suggested that infection rates are low and that there are few long-term complications associated with CVAD usage . We have reviewed 86 CVADs that have been inserted, since 1988, in 58 children with congenital bleeding disorders, aged 6 d to 16.5 years, attending Great Ormond Street Hospital, London, and the National Children's Hospital, Dublin . The devices have remained in situ for 2 weeks to 92 months (median 22.5 months) . Early (0-2 weeks) complications of CVAD insertion included nine bleeding episodes, one extravasation of factor concentrate, three allergic reactions to factor concentrate and five catheter infections . Overall, CVAD infection was the commonest problem encountered, with 52 devices (60%) becoming infected . Twenty-seven CVADs (31%) required removal . Infection rates in children without inhibitors (29/68) were 1/20 patient-months or 1 . 6 infections/1000 patient-days, but infection rates for those with inhibitors were 1/8.5 patient-months or 4.3/1000 patient-days . Staphylococcus epidermidis was the predominant organism (25/52) isolated . Blockage of CVAD (four) and catheter disconnection (four) were the most frequently occurring non-infectious long-term complications . Skin erosion of the port was also seen in three children, in one child at 20 months, in one at 29 months and in one at 34 months after insertion . This study demonstrates a high CVAD infection rate and highlights the long-term complications of CVAD usage. Ann Thorac Surg, 2000 Aug, 70(2), 547 - 52 Mediastinal false aneurysm after thoracic aortic surgery; Katsumata T et al.; BACKGROUND: Postoperative mediastinal false aneurysm is associated with a substantial morbidity and mortality . Surgical treatment is mandatory, although the individual approach varies according to the type of pathologic process, infection status, and site of origin of the aneurysm . METHODS: Between April 1993 and February 1999, we treated 10 patients, aged 25 to 73 years, with anastomotic mediastinal false aneurysm originating from the proximal thoracic aorta . Nine had undergone prior operations on the ascending aorta (7, type A dissection repair; 1, aortitis; 1, root abscess) with a Dacron conduit (n = 5) or valved conduit (n = 4) . The last patient had undergone valve replacement for excavating aortic root sepsis . False aneurysms were detected from 2 to 70 months after the most recent operation . Three patients had positive tissue cultures . The surgical procedure was direct suture repair of the disrupted anastomosis in 5, root or ascending aortic replacement with an aortic homograft in 4, and Dacron graft interposition in 1 . Hypothermic low-flow perfusion with or without circulatory arrest was used in all patients . RESULTS: There was one hospital death caused by staphylococcal mediastinitis . A false aneurysm recurred after direct suture repair in 2 patients with underlying type A dissection or aortitis . This resulted in one late death . One patient experienced a neurologic event during removal of an infected vascular graft . All 8 surviving patients are alive and well after a mean follow-up of 2 years . Three patients with mycotic false aneurysms remain free from infection after aortic homograft replacement . CONCLUSIONS: Mediastinal false aneurysms are surgically taxing . Low-flow hypothermic perfusion with or without circulatory arrest allows safe reentry . Radical surgery provides a satisfactory outcome in infected patients . Local repair of suture dehiscence in pathologic tissues may predispose to recurrence . We suspect that excessive use of formalin in gelatin-resorcin-formol glue may predispose to tissue necrosis. Biophys J, 2000 Sep, 79(3), 1610 - 20 High apparent dielectric constants in the interior of a protein reflect water penetration; Dwyer JJ et al.; A glutamic acid was buried in the hydrophobic core of staphylococcal nuclease by replacement of Val-66 . Its pK(a) was measured with equilibrium thermodynamic methods . It was 4.3 units higher than the pK(a) of Glu in water . This increase was comparable to the DeltapK(a) of 4.9 units measured previously for a lysine buried at the same location . According to the Born formalism these DeltapK(a) are energetically equivalent to the transfer of a charged group from water to a medium of dielectric constant of 12 . In contrast, the static dielectric constants of dry protein powders range from 2 to 4 . In the crystallographic structure of the V66E mutant, a chain of water molecules was seen that hydrates the buried Glu-66 and links it with bulk solvent . The buried water molecules have never previously been detected in >20 structures of nuclease . The structure and the measured energetics constitute compelling and unprecedented experimental evidence that solvent penetration can contribute significantly to the high apparent polarizability inside proteins . To improve structure-based calculations of electrostatic effects with continuum methods, it will be necessary to learn to account quantitatively for the contributions by solvent penetration to dielectric effects in the protein interior. Arq Bras Cardiol, 2000 Apr, 74(4), 324 - 8 Peripheral arterial embolism . Report of hospitalized cases; Pereira Barretto AC et al.; OBJECTIVE: We analyzed the frequency of peripheral embolisms, the underlying heart disease,triggering factors, the sites of the emboli, and evolution of the patients . METHODS: We analyzed 29 cases of peripheral arterial embolism out of a total of 20,211 hospitalizations in a cardiology center in the city of Sao Paulo . The age was 51.89+/-18.66 years, and 15 were males . RESULTS: Embolism in the right lower limb occurred in 18 patients (62.0%),in the left lower 11(37.9%) and right upper 3 (10.3%) limbs, and in the left arm (1) . Four patients had embolism in two limbs . The heart disease, mitral valvar heart disease (9 patients - 31.0%); infective endocarditis (7- 24.1%); dilated cardiomyopathy (6 - 20.6%); ischemic coronary heart disease (6 patients - 20.6%); and one patient with cor pulmonale . Atrial fibrillation was observed in 20 patients (68.9%), chronic in 12 patients (41.3% ) and acute in 8 (27 . 5%) . All patients with mitral valvar heart disease had atrial fibrillation, chronic in 8 patients (88.8%); patients with cardiomyopathy and coronary heart disease, 4 in each group had atrial fibrillation, acute in 60% of the patients.Patients with infective endocarditis, 3 had staphylococcus and 2 Gram-negative bacteria . In the follow-up, 2 patients (6.8%) required limbs amputation, and 5 (17.2%) died due to embolism . CONCLUSION: Most of the time, embolism does not cause permanent complications . Our data highlight the importance of anticoagulation for patients acute atrial fibrillation in myocardial dysfunction and for patients with chronic atrial fibrillation in cases of mitral valvar heart disease to prevent peripheral embolism. Blood, 2000 Sep 1, 96(5), 1853 - 6 Kinetics of CXCR4 and CCR5 up-regulation and human immunodeficiency virus expansion after antigenic stimulation of primary CD4(+) T lymphocytes; Maier R et al.; The chemokine receptors CCR5 and CXCR4 are coreceptors for the human immunodeficiency virus (HIV) and determine the cell tropism of different HIV strains . Previous studies on their regulation were performed under conditions of unspecific T-lymphocyte stimulation and provided conflicting results . To mimic physiologic conditions, highly purified primary Staphylococcus enterotoxin B (SEB)-reactive CD4 T lymphocytes were stimulated in the presence of autologous antigen-presenting cells and the kinetics of CCR5 and CXCR4 surface expression and HIV replication were studied . Both chemokine receptors were transiently up-regulated with maximal expression at day 3 after stimulation . The stimulated T cells were equally susceptible to productive infection with R5-and X4-tropic virus strains . Thus, antigenic stimulation of T cells promotes efficient replication of both, T cell-tropic and macrophage-tropic HIV . (Blood . 2000;96:1853-1856) Hepatology, 2000 Sep, 32(3), 507 - 13 Unresponsiveness of intrahepatic lymphocytes to bacterial superantigen: rapid development of suppressive Mac-1(high) cells in the mouse liver; Terabe M et al.; We previously found that a small dose (2 microg per mouse) of staphylococcal enterotoxin B (SEB) induced early emerging unresponsiveness in intrahepatic-lymphocyte populations (IHLs) . The purpose of this study was to reveal the inducing role of accessory cells involved in IHLs in this phenomenon . IHLs prepared at 3 to 24 hours after SEB injection failed to proliferate in response not only to SEB but also to SEA, representing ligand-nonspecific unresponsiveness, whereas spleen cells (SPCs) and mesenteric lymph-node cells showed transient proliferation . Unresponsiveness in IHLs was related to a deficit of their accessory cell function as measured by coculture of irradiated IHLs and antigen-specific, type 1 T-helper (Th1) clone cells . High levels of nitrite were detected in the culture supernatant . Supplement of N(G)-monomethyl-L-arginine lowered nitrite levels and concurrently restored the proliferative response of Th1 cells, indicating the involvement of nitric oxide in suppression . Adherent cells prepared from IHLs well reproduced these results . As shown by flow cytometry, Mac-1(high) Ia(+) cells, which mainly included F4/80(+) cells (macrophages) and a minor population of CD11c(+) cells (dendritic cells), increased in proportion in IHLs but not in SPCs at 6 to 24 hours . Depletion of Mac-1(high) cells from IHLs with antibody-coated magnetic beads recovered the proliferative response . Depleted Mac-1(high) cells had a monocytoid appearance . In immunostained sections, Kupffer cells came to highly express both Mac-1 and Ia at 12 hours . These results indicate that Mac-1(high)Ia(+) adherent cells, largely Kupffer cells activated by SEB, nonspecifically suppress the proliferation of Th1 cells via nitric oxide production before manifestation of ligand-specific unresponsiveness. Antimicrob Agents Chemother, 2000 Sep, 44(9), 2530 - 3 Identification of a plasmid-borne chloramphenicol-florfenicol resistance gene in Staphylococcus sciuri; Schwarz S et al.; The 16.5-kbp plasmid pSCFS1 from Staphylococcus sciuri mediated combined resistance to chloramphenicol and florfenicol . The gene responsible for this resistance property, cfr, was cloned and sequenced . The amino acid sequence of the Cfr protein revealed no homology to known acetyltransferases or efflux proteins involved in chloramphenicol and/or florfenicol resistance or to other proteins whose functions are known. Antimicrob Agents Chemother, 2000 Sep, 44(9), 2286 - 90 A single oral dose of thalidomide enhances the capacity of lymphocytes to secrete gamma interferon in healthy humans; Verbon A et al.; Thalidomide is increasingly being used as adjuvant therapy for patients with mycobacterial and human immunodeficiency virus (HIV) infections . The T-helper (Th)1 cytokine-Th2 cytokine balance critically determines the outcomes of these diseases . To obtain insight into the effect of thalidomide on the capacity of lymphocytes to produce Th1 and Th2 cytokines, six healthy volunteers received an oral dose (400 mg) of thalidomide . Before and at 3, 6, and 24 h after ingestion of thalidomide, peripheral blood mononuclear cells (PBMCs) were isolated and stimulated for 24 h with the T-cell stimulant staphylococcal enterotoxin B (SEB) or anti-CD3/CD28 . In all six volunteers ingestion of thalidomide was associated with enhanced SEB- and anti-CD3/CD28-induced production of the Th1 cytokine gamma interferon (P < 0.05) and a decrease in the level of anti-CD3/CD28-induced interleukin-5 (IL-5) production (P < 0.05) . The levels of IL-2 (Th1) and IL-4 (Th2) released remained unchanged . These changes were accompanied by an increase in the amount of IL-12p40 released by the PBMCs 6 h after ingestion of thalidomide (P < 0.05) . Thus, a single oral dose of thalidomide causes a Th1-type response in healthy humans . This finding offers a potential explanation for the positive effect of thalidomide in patients with mycobacterial and HIV infections. Infect Immun, 2000 Sep, 68(9), 5044 - 9 Purification of protease from a mixture of exfoliative toxin and newborn-mouse epidermis; Ninomiya J et al.; Although the role of exfoliative toxin in staphylococcal scalded-skin syndrome has been suggested to be that of a serine protease, it has not been demonstrated to show proteolytic activity . Our purpose was to purify a proteolytic enzyme from a mixture of exfoliative toxin and newborn-mouse epidermis . We used gel filtration and ion-exchange and hydroxyapatite chromatography with a high-pressure liquid chromatography system . A casein-hydrolyzing enzyme was isolated from the mixture . The molecular mass of the enzyme was confirmed to be 20 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis . Subcutaneous injection of the purified enzyme into newborn mice reproduced the epidermal splitting that is seen in staphylococcal scalded-skin syndrome . These results suggest that exfoliative toxin does not work as a protease itself but that some reaction between exfoliative toxin and an epidermal component(s) first produces a protease, after which epidermal splitting occurs. Biosens Bioelectron, 2000 Jan, 14(10-11), 795 - 804 Rapid detection and identification of biological and chemical agents by immunoassay, gene probe assay and enzyme inhibition using a silicon-based biosensor; Lee WE et al.; A rapid biosensor assay procedure that utilizes biotin streptavidin mediated filtration capture onto nitrocellulose membrane, in conjunction with a silicon-based light-addressable potentiometric sensor (LAPS) was developed for detection and identification of biological and chemical threat agents . Sandwich immunoassays, nucleic acid hybridization assays and enzyme inhibition assays are described . For immunoassays, the lower limits of detection (LOD) per 100-microl sample were approximately 5 pg/ml for protein (Staphylococcal enterotoxin B), 2 ng/ml for virus (Newcastle disease virus), and 20 ng/ml for vegetative bacteria (Brucella melitensis) . In a dual gene probe assay format, the LOD was 0.30 fmol (1.8 x 10(8) copies per 60-microl) of single stranded target DNA . Enzyme inhibition assays on the LAPS using acetylcholinesterase were able to detect soman and sarin in aqueous samples at 2 and 8 pg (100 and 600 pM), respectively . The assays were easy to perform and required a total time equal to the reaction period plus about 15 min for filtering, washing and sensing . The assay format is suitable for detection of a wide range of infectious and toxic substances . New assays can be developed and optimized readily, often within 1 or 2 days. Biosens Bioelectron, 2000 Jan, 14(10-11), 785 - 94 Array biosensor for detection of biohazards; Rowe-Taitt CA et al.; A fluorescence-based biosensor has been developed for simultaneous analysis of multiple samples for multiple biohazardous agents . A patterned array of antibodies immobilized on the surface of a planar waveguide is used to capture antigen present in samples; bound analyte is then quantified by means of fluorescent tracer antibodies . Upon excitation of the fluorophore by a small diode laser, a CCD camera detects the pattern of fluorescent antibody:antigen complexes on the waveguide surface . Image analysis software correlates the position of fluorescent signals with the identity of the analyte . This array biosensor has been used to detect toxins, toxoids, and killed or non-pathogenic (vaccine) strains of pathogenic bacteria . Limits of detection in the mid-ng/ml range (toxins and toxoids) and in the 10(3)-10(6) cfu/ml range (bacterial analytes) were achieved with a facile 14-min off-line assay . In addition, a fluidics and imaging system has been developed which allows automated detection of staphylococcal enterotoxin B (SEB) in the low ng/ml range. Biosens Bioelectron, 2000 Jan, 14(10-11), 779 - 84 Optical flow-cell multichannel immunosensor for the detection of biological warfare agents; Koch S et al.; An automated optical flow cell multichannel immunosensor for the detection and identification of toxins, viruses and bacterial particles is presented . A solid phase ELISA, based on a peroxidase label for signal generation and on fused silica capillaries as a support for immobilized antibodies, has been employed for analyte detection and identification . The sensing and signal transducing component of the sensor consists of a light-emitting diode and a photodetector . The device is fitted with three channels allowing the simultaneous detection of three agents . An integrated flow injection analysis system ensures automation of the assay cycles . Data on the detection of the bacterial toxin staphylococcal enterotoxin B (SEB), the bacteriophage M13 as a viral agent, and Escherichia coli as a bacterial agent are presented. J Vet Med Sci, 2000 Jul, 62(7), 757 - 8 Ultrasonographic findings of an ovarian abscess in a cow; Zulu VC et al.; Ultrasonographic examination was carried out in a cow with an ovarian abscess . Real time ultrasound scanning using a 7.5 MHz linear probe per rectum revealed an active left ovary with follicles of up to 11 mm in diameter and an enlarged right ovary measuring 6 x 6 cm, with a mass of 4 x 3 cm . The mass had varied echotexture, a thick hyperechoic boundary and a hypoechoic centre with echoic foci . It was tentatively diagnosed as an ovarian tumour or abscess . After ovariectomy, grossly the right ovary measured 6 x 6 x 5 cm and on cutting cheesy pus oozed out . The abscess was completely localised within the ovary, 4 cm in diameter and encapsulated thickly . On bacteriological examination Staphylococcus spp . was isolated . This is a rare case of ovarian abscess. Dtsch Med Wochenschr, 2000 Jul 14, 125(28-29), 862 - 5 {Late spreading of tuberculosis in tubercular spondylitis}; Herget GW et al.; HISTORY AND ADMISSION DIAGNOSIS: A 65-year-old man, known to have had a gastric ulcer and chronic rheumatoid arthritis as well as alcohol and nicotine abuse, was admitted because of suspected endocarditis . Physical examination revealed marked pain on pressure over the throacic spine . Vesicular breath sounds were reduced over the entire thorax and there was a systolic murmur over Erb's point (above the right clavicle) . There was a purulent bursitis over the olecranon . INVESTIGATIONS: Abnormal laboratory tests were: elevated C-reactive protein, elevated leucocyte count (up to 33 thousand during the hospital stay) . Smears from the bursitis and blood cultures revealed Staph . aureus . Computed tomography demonstrated a fracture of the 7th thoracic vertebra with a paravertebral abscess . Echocardiography showed anatherosclerotic aortic valve with floating particles . DIAGNOSIS, TREATMENT AND COURSE: Treatment of the suspected staphylococcal bacteraemia with purulent bursitis, spondylitis and aortic valvar endocarditis was begun with broad-spectrum antibiotics, but the patient soon developed a severe acute respiratory distress syndrome and he died of multi-organ failure . Autopsy revealed as cause of death left heart failure with aortic valvar endocarditis and gelatinour pneumonia caused by late tubercular dissemination from the tubercular spondylitis . CONCLUSION: Tuberculosis can be a life-threatening infection . Uncharacteristic history and extrapulmonary manifestations can make it very difficult to arrive at the correct diagnosis. Eur J Immunol, 2000 Jul, 30(7), 2048 - 55 Beta-chemokines inhibit activation-induced death of lymphocytes from HIV-infected individuals; Pinto LA et al.; The present study investigates the role of the HIV-suppressive beta-chemokines macrophage inflammatory protein (MIP)-1alpha, MIP-1 and RANTES in activation-induced cell death (AICD) . A pool of these beta-chemokines reduced anti-CD3-induced apoptosis of T cell blasts from healthy blood donors in a dose-dependent manner . Although the pooled beta-chemokines were more effective, the inhibitory effect could also be mediated by each of the individual chemokines and was blocked by neutralizing anti-chemokine antibodies . The beta-chemokines also inhibited pokeweed mitogen/staphylococcal enterotoxin B-induced T lymphocyte apoptosis in 33/49 HIV-infected (HIV+) individuals . This anti-apoptotic effect was not correlated with the patients' CD4 T cell counts . beta-chemokines did not lead to altered secretion of IL-2, IL-4, IFN-gamma or IL-10 in response to activation stimuli in either normal T cell blasts or peripheral blood mononuclear cells from HIV+ individuals . Co-incubation with beta-chemokines did not inhibit anti-CD3-induced expression of cell surface Fas ligand, nor did it alter levels of the death receptor Fas or Bcl-2 in T cell blasts, suggesting that the beta-chemokines are blocking AICD downstream of Fas . These observations indicate that beta-chemokines may play a novel role as modulators of AICD, in addition to their known role as chemoattractants and inhibitors of HIV replication. Chin J Biotechnol, 1999, 15(3), 177 - 82 A new system for expressing heterologous gene in Escherichia coli regulated by oxygen consistence in the environment; Tong Q et al.; The expression of Vitreosilla hemoglobin gene (vgb) is regulated by oxygen consistence in E . coli . The gene transcription is activated under the condition of limited oxygen . A new system for expressing heterologous gene in E . coli regulated by dissolved oxygen consistence was constructed . It includes a host bacteria GJ100, which contained T7 RNA polymerase gene controlled by vgb promoter, and an expression vector on which the heterologous gene was under the control of a T7 promoter . The results indicated that E . coli thioredoxin A, IgG binding domain of Staphylococcus protein A(ZZ), snake neurotoxin, salmon calcitonin hexa-polymer, human interleukinII (IL2) and human pro-urokinase genes could be expressed efficiently . Expression level was more than 30% of the total cellular protein. J Pharm Biomed Anal, 2000 Aug 15, 23(2-3), 363 - 74 Determination of slime-producing S . epidermidis specific antibodies in human immunoglobulin preparations and blood sera by an enzyme immunoassay: correlation of antibody titers with opsonic activity and application to preterm neonates; Lamari F et al.; Slime-producing Staphylococcus epidermidis is responsible for severe infections in immunocompromised patients and, particularly, in premature infants who are transiently deficient in IgG . A sulfated polysaccharide with molecular mass of 20-kDa (20-kDa PS) has been recognized as the major polysaccharide component and antigenic determinant of S . epidermidis extracellular slime layer . The presence of adequate amounts of antibodies to 20-kDa PS in patients' sera would be of importance to prevent or treat slime-producing S . epidermidis bacteremia . Administration of intravenous immunoglobulin (IVIG) is considered to be a reasonable IgG replacement therapy and has been widely used to prevent or treat neonatal sepsis . Clinical trials have shown conflicting results on the efficacy of IVIGs and this phenomenon has been attributed to the variability of IVIG preparations in the content and opsonic activity of IgG against microorganisms of clinical importance . Monitoring of antibodies to distinct bacterial macromolecules, which are species-specific and responsible for bacterial infections, has not been performed previously . A highly precise and repeatable enzyme immunoassay was developed to determine quantitatively the levels of antibodies against the 20-kDa PS of S . epidermidis slime . The amount of 20-kDa PS specific antibodies found in 27 lots of an IVIG preparation (Sandoglobulin) correlated well with their in vitro opsonic activity against slime-producing S . epidermidis . The majority of lots (75%) having titers higher than 200 units/ml showed significant opsonic activity (50-75%) towards slime-producing S . epidermidis . Sandoglobulin lots with titers higher than 200 units/ml of 20-kDa PS specific IgG were administered as a prophylactic agent to low-birth weight (lower than 1700 g) preterm neonates immediately after birth . The levels of total and 20-kDa PS specific IgG in neonates' blood sera were significantly higher than those found in the control group, even 10 days after the last infusion . The rate of slime-producing S . epidermidis bacteremia in neonates who received IVIG was also considerably lower than those in the control group . The results of this study suggest that specific IgG titers estimated by the developed enzyme immunoassay may well be indicative of the IVIG opsonic activity against slime-producing S . epidermidis . Furthermore, administration of Sandoglobulin with titers higher than a cut-off value of 200 units/ml may significantly protect preterm neonates against slime-producing S . epidermidis bacteremia. Rev Latinoam Microbiol, 1999 Apr-Jun, 41(2), 53 - 7 {Dichotomous keys for identifying Staphylococcus isolated from bovine milk}; Demo M et al.; Many efforts have been done in order to identify species of Gram positive and catalase positive coccus isolated from raw milk . The aim of this epidemiological observation, was to identify different strains isolated from raw milk according to conventional criteria and to recommend a useful and practical methodology for their correct identification . Gram positive and catalase positive coccus were tested for bacitracin resistance, novobiocin susceptibility, coagulase and others tests (e.g . carbohydrate fermentation) . Dichotomic keys were designed to typify Staphylococcus' species . They were grouped into: Group I, coagulase positive, novobiocin susceptibility; Group II coagulase negative novobyocin resistance; and Group III coagulase negative novobiocin susceptibility . 62 strains were tested and S . hycus (17.7%) was the most isolated species, S . aureus (14.5%), S . sciuri (12.9%) and S . haemolyticus (11.3%) were also isolated . S . epidermidis was not isolated. Rev Latinoam Microbiol, 1998 Jan-Jun, 40(1-2), 45 - 52 Arsenic resistance determinants from environmental bacteria; Esquivel JA et al.; Arsenic resistance determinants from 42 environmental bacterial isolates (32 Gram negative) were analyzed by DNA: DNA hybridization using probes derived from Escherichia coli and Staphylococcus plasmid or chromosomal arsenic resistance (ars) genes . In colony hybridization assays, 11 and 1 Gram negative strains hybridized with the E . coli chromosome and plasmid probes, respectively . No hybridization was detected using a probe containing only the arsA (ATPase) gene from E . coli plasmid or with a Staphylococcus plasmid ars probe . From Southern hybridization tests of some of the positive strains it was concluded that homology to ars chromosomal genes occurred within chromosome regions, except in an E . coli isolate where hybridization occurred in both the chromosome and a 130-kb plasmid . Our results show that DNA sequences homologous to E . coli ars chromosomal genes are commonly present in the chromosomes of environmental arsenic-resistant Gram negative isolates. Rev Latinoam Microbiol, 1997 Jan-Jun, 39(1-2), 57 - 63 Prevention of nonspecific reactions on reversed passive latex agglutination assay (RPLA) for detecting low amounts of staphylococcal enterotoxins; Pereira ML et al.; The SET-RPLA, from Denka Seiken Co . Ltd., Tokio, a commercial reversed passive latex agglutination test kit, has been recommended to establish the enterotoxicity capacity of some staphylococcal strains, implicated in food poisoning outbreaks that produce low levels of enterotoxins (SE) . Despite the RPLA specificity, the occurrence of nonspecific reactions when testing low-SE-producing is common . In order to control these nonspecific reactions the addition of purified normal rabbit IgG purified was applied on approximately 350 staphylococcal isolates from human milk and anatomic sites of healthy dental student carriers . The results indicated that addition of 5% (v/v) of purified normal rabbit IgG (0.74 mg/mL) to the culture supernatant fluid is a simple and reliable tool for the controlling of nonspecific reactions in the RPLA assay. Clin Exp Immunol, 2000 Aug, 121(2), 181 - 6 Staphylococcal toxin-induced T cell proliferation in atopic eczema correlates with increased use of superantigen-reactive Vbeta-chains in cutaneous lymphocyte-associated antigen (CLA)-positive lymphocytes; Davison S et al.; Staphylococcal superantigens have been implicated in the pathogenesis of atopic dermatitis (AD) . This may occur through superantigenic activation of T lymphocytes and their subsequent induction of the skin homing receptor CLA on activated cells . We investigated the proliferative responses of peripheral blood mononuclear cells (PBMC) from 10 patients with an infective exacerbation of AD and six normal controls to the staphylococcal superantigens, staphylococcal enterotoxin A and B (SEA, SEB) and toxic shock syndrome toxin-1 (TSST-1), and the mitogens phytohaemagglutinin (PHA) and concanavalin A (Con A) . We also assessed CLA and T cell receptor (TCR) Vbeta-chain expression by immunofluorescence and flow cytometry before and after stimulation . PBMC from AD patients showed two-fold increased proliferation to SEA and SEB (P < 0.01) compared with normals, whereas the response to mitogenic stimulation was identical . Analysis of (TCR) Vbeta-chain expression demonstrated increased use of superantigen-reactive Vbeta families in freshly isolated PBMC in AD patients compared with controls . This pattern of Vbeta-chain expression was only observed in the CLA+ but not the total population of T cells . Furthermore, there was a positive correlation between the enhanced PBMC proliferative response and increased expression of superantigen-reactive Vbeta families in atopic patients . These data support the concept that superantigens are important in the pathogenesis of this common condition, and also provide evidence that the increased use of certain Vbeta families in circulating, CLA+, skin homing lymphocytes is of functional significance. J Radiol, 2000 Mar, 81(3 Suppl), 392 - 408 {Imaging of chronic hip pain in adults}; Chevrot A et al.; Adult hip pathologies are mainly represented by the degenerative disease, so called "osteoarthrosis, or more precisely coxarthrosis" . The means of imaging are exposed, according to their specific value: X Rays (measurement of the characteristic angles of the adult hip), Arthrography, CT Scanner, Arthro-CT Scanner, MRI, Bone Scintigraphy, Ultrasonography . Clinical findings differentiate a mechanical syndrome and an inflammatory syndrome . The coxarthrosis is the most frequent, under two forms: primary (idiopathic) coxarthrosis and secondary coxarthrosis . Primary (idiopathic) coxarthrosis has a localised narrowing of the joint space, osteophyte formation, subchondral sclerosis, cyst formation . The destruction progresses slowly, in 10 to 15 years leading to a complete destruction . Bilaterality is frequent . it is treated with total hip prosthesis . There is a rapid form (1 to 2 years) (Postel's Disease) . Secondary coxarthrosis occurs after architectural vice, chondral diseases, lack of balance between the size of the head and the acetabulum as in the case of previous fracture or dislocation, avascular bone necrosis of the head of the femur, Paget's disease . Calcium pyrophosphate Deposition disease (CPPD) involves mostly aged women, and also leads to cox-arthrosis . Avascular bone necrosis of the head of the femur involves young adults . Bilateral involvement are frequent . MRI is the most sensitive and the most specific means of early diagnosis, The area of bone necrosis appears as well defined modifications of the upper head of the femur, precisely surrounded by a low signal intensity line on both Ti and T2 weighted imaging . MRI shows articular effusion, bone marrow edema . Scintigraphy gives early findings which are a characteristic, but non specific, hot spot . CT scanner is used for hip destruction evaluation . o Algodystrophy: transient osteoporosis of the hip has a cyclic course, lasting 3 to 9 months . MRI shows an inflammatory pattern in the area of the process(dark in Ti and white in T2, with positive Gadolinium response) . Scintigraphy is positive . Staphylococcus location in the hip can be acute or chronic . MRI shows joint effusion, cystic formation and subchondral non specific modifications . Tuberculosis of the hip joint is relatively rare . Greater trochanteric tuberculous involvement is possible under special contexts . Chronic Inflammatory diseases are represented by Rheumatoid Arthritis, Spondylarthritis and other chronic inflammatory diseases . Synovial tumors such as Pigmented Villo Nodular Synovitis, Primary Osteochondromatosis, synovial sarcoma have special presentations . The subchondral bone can be involved by amorphous depositions such as in tophaceous gout, different varieties of lipidosis, amyloidosis, reticulo histiocytosis . Pen arthropathies are enthesopathies in the anterior rectus tendon, calcifying tendonitis (not to be confused with calcifying soft tissue tumor/chondrosarcoma) . The pelvis bone and the femur are involved by primary and secondary tumors or by insufficiency fractures which can mislead to hip pathologies. Knee, 2000 Jul 1, 7(3), 171 - 174 Infected knee treated by total knee arthroplasty; Mirza AH et al.; The perceived wisdom in orthopaedics precludes insertion of implants in cases of post-recent active joint sepsis . However, revision knee surgery after infected arthroplasty has now been recognised to be safe and efficacious in specialised centres . We illustrate the concept of using principles of revision surgery to successfully treat knees destroyed by primary staphylococcal septic arthritis. J Mol Biol, 2000 Aug 11, 301(2), 247 - 56 Ensemble modulation as an origin of denaturant-independent hydrogen exchange in proteins; Wooll JO et al.; Native state hydrogen exchange (HX) has become a powerful tool for the analysis of conformational states that exist under native conditions . However, the interpretation of HX data in terms of conformational fluctuations is still controversial . In particular, it has been shown that many residues display exchange behavior that is independent of denaturant concentration . It has been postulated that this lack of denaturant dependence results from local fluctuations that do not expose appreciable amounts of buried surface area . Here, we use a general thermodynamic description of HX to explore the different possibilities for this behavior . We find that the denaturant dependence seen in HX experiments under native conditions is not a de facto indication of the amount of surface area exposure required for exchange . Instead, this behavior results from the relatively homogenous character of the conformational ensemble that exists under native conditions and the non-specific nature of denaturant effects . Furthermore, a comparison of the HX behavior from a stabilized mutant of Staphylococcal nuclease (SNase) with that predicted for the wild-type SNase from the COREX algorithm suggests that denaturant-independent exchange of many residues is consistent with significant (approximately 10 %) surface area exposure for this protein . J Immunol, 2000 Aug 15, 165(4), 1918 - 24 Helicobacter pylori urease binds to class II MHC on gastric epithelial cells and induces their apoptosis; Fan X et al.; Infection by Helicobacter pylori leads to injury of the gastric epithelium and a cellular infiltrate that includes CD4+ T cells . H . pylori binds to class II MHC molecules on gastric epithelial cells and induces their apoptosis . Because urease is an abundant protein expressed by H . pylori, we examined whether it had the ability to bind class II MHC and induce apoptosis in class II MHC-bearing cells . Flow cytometry revealed the binding of PE-conjugated urease to class II MHC+ gastric epithelial cell lines . The binding of urease to human gastric epithelial cells was reduced by anti-class II MHC Abs and by staphylococcal enterotoxin B . The binding of urease to class II MHC was confirmed when urease bound to HLA-DR1-transfected COS-1 (1D12) cells but not to untransfected COS-1 cells . Urease also bound to a panel of B cell lines expressing various class II MHC alleles . Recombinant urease induced apoptosis in gastric epithelial cells that express class II MHC molecules, but not in class II MHC- cells . Also, Fab from anti-class II MHC and not from isotype control Abs blocked the induction of apoptosis by urease in a concentration-dependent manner . The adhesin properties of urease might point to a novel and important role of H . pylori urease in the pathogenesis of H . pylori infection. Surgery, 2000 Aug, 128(2), 159 - 64 A central role for CD95 (Fas) in T-cell reactivity after injury; Kell MR et al.; BACKGROUND: Recent findings indicate that severe injury primes the immune system for an enhanced and lethal proinflammatory cytokine response against bacterial-derived superantigens . This study asked whether this response to injury involves the CD95 (Fas) signaling pathway . METHODS: To assess superantigen-mediated mortality, wild-type (WT) C57BL/6 and Fas-deficient C57BL/6 lpr (-/-) (lpr) mice underwent burn or sham injury and were challenged 2 hours later with staphylococcal enterotoxin B (SEB) . Spleen cells from sham and burn WT or lpr mice were stimulated in vitro with SEB to assess injury effects on IL-2, TNF-alpha, and IFN-gamma production . RESULTS: Lpr burn mice survived the SEB challenge (100% survival), while WT burn mice showed a high mortality (17% survival, P < 001, analysis of variance {ANOVA}) . Sham lpr or WT mice suffered no mortality to the SEB challenge . In vitro studies demonstrated that burn lpr mice produced significantly less TNF-alpha, IFN-gamma, IL-2 than burn WT mice (P <.01, ANOVA) . Burn injury markedly enhanced SEB-stimulated IFN-gamma production by WT spleen cells and CD8+ T cells, while this did not occur in SEB-stimulated lpr spleen cells . CONCLUSIONS: These findings support the hypothesis that the CD95 (Fas) signaling pathway plays an integral role in the injury-induced enhanced and lethal T-cell reactivity against bacterial superantigens. J Biomol NMR, 2000 Jun, 17(2), 137 - 51 RFAC, a program for automated NMR R-factor estimation; Gronwald W et al.; A computer program (RFAC) has been developed, which allows the automated estimation of residual indices (R-factors) for protein NMR structures and gives a reliable measure for the quality of the structures . The R-factor calculation is based on the comparison of experimental and simulated 1H NOESY NMR spectra . The approach comprises an automatic peak picking and a Bayesian analysis of the data, followed by an automated structure based assignment of the NOESY spectra and the calculation of the R-factor . The major difference to previously published R-factor definitions is that we take the non-assigned experimental peaks into account as well . The number and the intensities of the non-assigned signals are an important measure for the quality of an NMR structure . It turns out that for different problems optimally adapted R-factors should be used which are defined in the paper . The program allows to compute a global R-factor, different R-factors for the intra residual NOEs, the inter residual NOEs, sequential NOEs, medium range NOEs and long range NOEs . Furthermore, R-factors can be calculated for various user defined parts of the molecule or it is possible to obtain a residue-by-residue R-factor . Another possibility is to sort the R-factors according to their corresponding distances . The summary of all these different R-factors should allow the user to judge the structure in detail . The new program has been successfully tested on two medium sized proteins, the cold shock protein (TmCsp) from Termotoga maritima and the histidine containing protein (HPr) from Staphylococcus carnosus . A comparison with a previously published R-factor definition shows that our approach is more sensitive to errors in the calculated structure. Zhonghua Zhong Liu Za Zhi, 1997 May, 19(3), 188 - 91 {Mechanism and treatment of cancer cachexia in tumor-bearing mice}; Li T et al.; OBJECTIVE: To evaluate the role of TNF-alpha in the development of cancer cachexia, and the effects of TNF-alpha antibody and high agglutinated staphylococcin (BM828) in the treatment of experimental cancer cachexia . METHODS: T739 mice bearing LA795 tumor were used as murine tumor cachexia model and also the T739 receiving TNF-alpha . The level of serum TNF-alpha was assayed by TNF-alpha RIA in each group . Comparisons were made in the food intake and body weight of all the five groups including BM828 and TNF-alpha AB groups . RESULTS: Two weeks after being inoculated with LA795, the tumor-bearing mice developed significant anorexia and weight loss . The level of serum TNF-alpha in tumor-bearing mice was significantly higher than that in the non-tumor bearing mice (P < 0.01) . TNF-alpha administration to mice resulted also in marked anorexia and weight loss as in cancer cachexia . CONCLUSION: The results suggested that the mechanism of cachexia be correlated with TNF-alpha . BM828 and TNF-alpha AB have the potentiality of attenuating the development of cachexia in murine models. Rozhl Chir, 2000 Apr, 79(4), 162 - 4 {Autogenous bilateral aortofemoral reconstruction using the popliteal and superficial femoral veins}; Rezek Z; The author presents a case-history of a late infection of a bilateral aortofemoral bypass by a coagulase-negative staphylococcus and the definite solution by substitution of the contaminated vascular prosthesis by an anatomical autogenous bilateral bypass made from popliteal and superficial femoral veins . From the technical aspect the operation was very satisfactory without marked peroperative difficulties, the early postoperative course was also favourable, with readily palpable peripheral pulsations, without marked manifestations of venostasis on the lower extremities . From the third postoperative day onwards however progressive manifestations of multiorgan failure developed associated with septic shock and leading to a fatal outcome . Despite this the author considers in patients with a reasonable surgical risk this method of substitution of an infected vascular prosthesis in the aortofemoral region feasible with regard to its resistance to infection and long-term patency, though the operation is technically pretentious but easy to implement. J Laryngol Otol, 2000 May, 114(5), 366 - 9 Non-pseudomonal malignant otitis externa and jugular foramen syndrome secondary to cyclosporin-induced hypertrichosis in a diabetic renal transplant patient; Lancaster J et al.; We present the case of a 58-year-old diabetic renal transplant patient who developed a left jugular foramen syndrome, secondary to an ipsilateral staphylococcal malignant (necrotizing) otitis externa . This followed a protracted episode of uncomplicated otitis externa with no evidence of bone involvement on computed tomography (CT) scanning . Cyclosporin-induced hypertrichosis (excess hair growth) in his external auditory canal contributed greatly to the initial difficulty in managing his otitis externa . Following an initial successful treatment with prolonged intravenous antibiotics the patient relapsed with a secondary infection in the same anatomical site due to Candida parapsilosis . Despite further intensive treatment including antimicrobials, a subtotal petrosectomy and hyperbaric oxygen therapy he eventually succumbed to his disease. Harefuah, 1998 Jul, 135(1-2), 15 - 6, 87 {Discitis in brucellosis is not responsive to anti-brucella treatment}; Heldenberg D et al.; A 14-year-old girl with fever and low back pain was diagnosed as having brucellosis and diskitis . Treatment with doxycycline and gentamicin had no effect nor was there any improvement when the treatment was changed to doxycycline and rifampicin . With cessation of this treatment and administration of cloxacillin fever was reduced and the back pain disappeared . After 12 days of i.v . cloxacillin, she was released from hospital with instructions to continue cloxacillin orally for another 4 weeks . We conclude that when a patient is suffering from brucellosis and diskitis and does not respond to the usual anti-brucellosis treatment, an anti-staphylococcal preparation should be added. Toxicol Appl Pharmacol, 2000 Aug 1, 166(3), 214 - 21 CTL hyporesponsiveness induced by 2,3,7, 8-tetrachlorodibenzo-p-dioxin: role of cytokines and apoptosis; Prell RA et al.; Studies have shown that blocking B7-mediated costimulation induces T cell tolerance via anergy or apoptosis . Provision of exogenous IL-2 can reverse or prevent the induction of tolerance . We have previously shown that TCDD-induced suppression of the CTL response to allogeneic P815 tumor cells is accompanied by decreased expression of CD86 (B7-2) as well as suppressed IL-2 and IFNgamma production . In the present studies, the role of IL-2 and IFNgamma and the analysis of inappropriate deletion of CD8(+) cells was examined . Administration of IL-2 on days 7-9 relative to the injection of P815 tumor cells dose-dependently increased the CTL activity and the generation of CD8(+) CTL effector cells in TCDD-treated mice . This increased CTL response was not due to recruitment of naive CTL precursors (CTLp), suggesting that a small pool of activated CTLp in TCDD-treated mice could respond to the IL-2 . A much larger pool of activated CTLp in control mice was also expanded by IL-2 treatment . In contrast, treatment with IFNgamma during the same time period did not alter CTL activity in control or TCDD-treated mice . To address the possibility that insufficient IL-2 early in the response was responsible for the reduced pool of activated CTLp in TCDD-treated mice, IL-2 was administered on days 1-3 after P815 injection . However, not only did early treatment with IL-2 fail to restore the response in TCDD-treated mice, it suppressed the CTL response of non-TCDD-treated mice . To test whether exposure to TCDD induced apoptosis of activated CD8(+) T cells, phosphatidylserine (PS) expression was measured on various days after P815 tumor challenge . Surprisingly, the percentage of apoptotic CD8(+) T cells was significantly lower in TCDD-treated mice compared to controls throughout the allograft response . Similarly, exposure to TCDD failed to enhance peripheral deletion of Vbeta3(+)CD8(+) T cells after injection of the superantigen Staphylococcal enterotoxin A (SEA) . Taken together, the data indicate that TCDD induces an early defect in CTLp activation that is not due to insufficient IL-2 or deletion of CD8(+) cells and may implicate a novel mechanism by which ligands of the Ah receptor disrupt CTL precursor activation . J Virol, 2000 Aug, 74(16), 7442 - 50 Prolonged dominance of clonally restricted CD4(+) T cells in macaques infected with simian immunodeficiency viruses; Chen ZW et al.; The repertoire of functional CD4(+) T lymphocytes in human immunodeficiency virus type 1-infected individuals remains poorly understood . To explore this issue, we have examined the clonality of CD4(+) T cells in simian immunodeficiency virus (SIV)-infected macaques by assessing T-cell receptor complementarity-determining region 3 (CDR3) profiles and sequences . A dominance of CD4(+) T cells expressing particular CDR3 sequences was identified within certain Vbeta-expressing peripheral blood lymphocyte subpopulations in the infected monkeys . Studies were then done to explore whether these dominant CD4(+) T cells represented expanded antigen-specific cell subpopulations or residual cells remaining in the course of virus-induced CD4(+) T-cell depletion . Sequence analysis revealed that these selected CDR3-bearing CD4(+) T-cell clones emerged soon after infection and dominated the CD4(+) T-cell repertoire for up to 14 months . Moreover, inoculation of chronically infected macaques with autologous SIV-infected cell lines to transiently increase plasma viral loads in the monkeys resulted in the dominance of these selected CDR3-bearing CD4(+) T cells . Both the temporal association of the detection of these clonal cell populations with infection and the dominance of these cell populations following superinfection with SIV suggest that these cells may be SIV specific . Finally, the inoculation of staphylococcal enterotoxin B superantigen into SIV-infected macaques uncovered a polyclonal background underlying the few dominant CDR3-bearing CD4(+) T cells, demonstrating that expandable polyclonal CD4(+) T-cell subpopulations persist in these animals . These results support the notions that a chronic AIDS virus infection can induce clonal expansion, in addition to depletion of CD4(+) T cells, and that some of these clones may be SIV specific. Antibiot Khimioter, 2000, 45(6), 10 - 2 {The antimicrobial activity and acute toxicity of the polymer salts of gentamycin}; Solovskii MV et al.; By neutralization of copolymers of N-(2-hydroxypropyl) methacrylamide with acrylic acid and copolymers of N-vinylpyrrolidone with crotonic and p-crotonoylaminophenoxyacetic acids in the presence of gentamycin, water-soluble polymer salts containing from 10 to 25 mass% of gentamycin were obtained . These salts regardless of gentamycin content completely retain high level of antimicrobial activity of gentamycin against Staphylococcus spp . and Escherichia coli and are characterized by less (by more than one order of magnitude) acute toxicity. Aust N Z J Surg, 2000 Jul, 70(7), 480 - 4 Contamination of banked femoral head allograft: incidence, bacteriology and donor follow up; Sommerville SM et al.; BACKGROUND: Allograft donations are not uncommonly found to be contaminated . The issue of contaminated donations from live donors at the time of surgery, and the significance of this to the patient in terms of subsequent sepsis of the arthroplasty, were examined . METHODS: The donations of femoral heads to the Queensland Bone Bank over a 9-year period were reviewed, and the incidence and bacteriology of contamination were detailed . Clinical outcomes were determined for donors who had positive cultures at the time of retrieval and they were compared with those of culture-negative donors . RESULTS: Between March 1987 and February 1996, 232 femoral heads were donated to the Queensland Bone Bank . Four specimens were sent for culture with each femoral head (surface swab of femoral head, acetabular swab, bone biopsy and capsule) . In 51 cases, one or more positive cultures were obtained (22% contamination rate) . The majority of orga