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Selected AbstractsClinical evaluation of resilient denture liners.JOURNAL OF PROSTHODONTICS, Issue 3 2003Part 2: candida count, speciation Purpose The purpose of this study was to count and to speciate Candida isolated from 2 resilient denture liners, Molloplast-B and MPDS-SL. Materials and Methods A group of 20 patients each had 1 maxillary denture and 2 mandibular dentures fabricated. One mandibular denture was lined with Molloplast-B, and 1 was lined with MPDS-SL. Each denture was used for 3 months. At the end of the 3-month period, the mandibular denture was surrendered, and a 5 × 5-mm circular resilient liner sample was obtained from the tissue surface of the lingual flange. Samples were processed, and Candida was isolated and counted. Speciation of Candida was performed using CHROMagar Candida and API 20C AUX strips. Results Molloplast-B had, on average, 5 times as many CFU/sample as MPDSL-SL, but this difference was not significant (p= 0.26). A sign test gave a similar nonsignificant trend (p= 0.057). CHROMagar identified several Candida species, and confirmation was made using API 20C AUX strips. One patient was lost to follow-up. Of 19 Molloplast-B samples, 7 had no growth, 4 grew C. albicans, 3 grew C. parapsilosis, 2 grew C. glabrata, 1 grew C. tropicalis, 2 grew a Trichosporon spp., and 2 grew a nonidentifiable colony. The analogous counts for 19 MPDS-SL samples were 10, 4, 1, 3, 0, 1, and 1 (p= 0.45 for culture positively, exact McNemar test). ConclusionsCandida growth on Molloplast-B was not significantly different from growth on MPDS-SL. Several yeast species were cultured from each material. The rates of culture-positive testing did not differ between the 2 resilient denture liners. [source] Significant cytotoxic activity in vitro of the EGFR tyrosine kinase inhibitor gefitinib in acute myeloblastic leukaemiaEUROPEAN JOURNAL OF HAEMATOLOGY, Issue 5 2008Elin Lindhagen Abstract Objectives:, Gefitinib inhibits epidermal growth factor receptor (EGFR) signalling, but may also act by non-EGFR dependent mechanisms. We have investigated the activity of gefitinib in haematological tumour cells, in particular acute myeloblastic leukaemia (AML). Methods:, Cytotoxic activity of gefitinib, alone or in combination with standard anti-leukaemic drugs, was assessed by the short-term fluorometric microculture cytotoxicity assay in tumour cells from 117 patients representing five haematological and five non-haematological malignancies. In AML, the EGFR status was analysed by immunochemistry. Gefitinib-induced apoptosis was investigated in a subset of AML samples, as well as in the leukaemia cell line MV-4-11, using a multiparametric high content screening assay. To confirm activation of caspase-3 in cells treated with gefitinib, a blocking test was carried out in which MV4-11 cells were pretreated with the specific caspase inhibitor DEVD-FMK. Results:, Gefitinib showed highest cytotoxic activity in AML (n = 19) with many samples being sensitive at concentrations achievable in clinical practice (<10 ,M), and no difference between previously untreated and relapsed patients. No correlation between the activity of gefitinib and standard antileukaemic drugs (cytarabine, doxorubicin, etoposide) was observed. Combining gefitinib with these drugs resulted in mainly additive or synergistic (etoposide) effects, with no evidence of sequence dependency. The AML cells did not express the EGFR. Gefitinib induced apoptosis, which was at least partly mediated by activation of the caspase-3 pathway. Conclusion:,In vitro, gefitinib has significant cytotoxic activity in AML by inducing apoptosis through non-EGFR dependent pathways. [source] Romarchite and associated phases as common corrosion products on pewter artifacts from marine archaeological sitesGEOARCHAEOLOGY: AN INTERNATIONAL JOURNAL, Issue 6 2004Stacie E. Dunkle Corrosion products were examined from typical pewter artifacts originating from six different submerged archaeological sites, dating to between ca. A.D. 1550 and 1733, along the eastern seaboard of North America and in the Caribbean Sea. The artifacts were viewed as 270,450-year long experiments revealing the phases and mechanisms of tin corrosion in seawater. All of the samples analyzed exhibit abhurite (Sn3O(OH)2Cl2), romarchite (SnO), and hydroromarchite (Sn3O2(OH)2) forming at the expense of the underlying artifact. Textural analysis suggests that abhurite is the first alteration product to form at the expense of the pewter; romarchite subsequently develops and then hydroromarchite. The outermost corrosion layers on several of the most corroded artifacts also exhibit cassiterite (SnO2) as a significant and apparently final phase to form during alteration. The absence of this mineral on many samples demonstrates that, while samples appeared to be stable under the conditions that were present, cassiterite had not yet had time to form. The very limited stability field for romarchite, based on data presented by Séby et al. (Geochimica et Cosmochimica Acta, 65, 3041,3053, 2001), suggests that its presence on these artifacts may be the result of a kinetic effect. The universal appearance of this mineral on corroding tin suggests that it is a required step in the oxidation of pure tin to the final most stable phase of cassiterite. The stability of romarchite and its effectiveness as an agent of passivation can provide insight into not only the formation of tin oxides but the rate of tin corrosion. This can have significant implications in the field of artifact preservation as well as more widespread industrial applications. © 2004 Wiley Periodicals, Inc. [source] HHV-6 A- or B-specific P41 antigens do not reveal virus variant-specific IgG or IgM responses in human serumJOURNAL OF MEDICAL VIROLOGY, Issue 3 2002Yunhe Xu Abstract The etiology of multiple sclerosis (MS) remains unknown, but there are indications of a role of human herpesvirus 6 (HHV-6), especially variant A, in the pathogenesis. Higher serum antibody reactivity against an HHV-6 early protein, p41, has been found in MS cases than in controls. The antigen, however, was purified from infected cells with a monoclonal antibody also reactive with a protein (p38) likely to be of cellular origin. To avoid serological crossreactivity with the cellular protein, recombinant p41 proteins from HHV-6A strain GS and HHV-6B strain Z29 were expressed as glutathione-S-transferase fusion proteins (p41-GST), and used as antigens in an enzyme-linked immunosorbent assay (ELISA). p41 variant specific monoclonal antibodies reacted strongly with the respective recombinant proteins. Serum IgM and IgG reactivities with the recombinant p41 antigens were analysed in patients with manifest MS, patients with optic neuritis, patients with other neurological diseases, and in one group of healthy controls. All sera were HHV-6 IgG seropositive by immunofluorescence. The serum IgM or IgG reactivities against the recombinant p41 antigens did not differ significantly between the groups, and the reactivities against the variant A and B antigens were identical. In many samples, the reactivity was very low. The results indicate that p41 is not an optimal target for HHV-6 serology studies, and that the data obtained with the p41 antigen prepared from infected cells (possibly including also p38) should be interpreted with caution. J. Med. Virol. 66:394-399, 2002. © 2002 Wiley-Liss, Inc. [source] Detection of Phytophthora nicotianae in Soil with Real-time Quantitative PCRJOURNAL OF PHYTOPATHOLOGY, Issue 1 2010Junli Huang Abstract Phytophthora nicotianae is one of the most important soil-borne plant pathogens. A rapid, specific and sensitive real-time polymerase chain reaction (PCR) detection method for P. nicotianae was established, which used primers targeting the internal transcribed spacer (ITS) regions of rDNA genes of Phytophthora spp. Based on the nucleotide sequences of ITS2 of 15 different species of Phytophthora, the primers and probe were designed specifically to amplify DNA from P. nicotianae. With a series of 10-fold DNA dilutions extracted from P. nicotianae pure cultures, the detection limit was 10 pg/,l in conventional PCR, whereas in SYBR Green I PCR the detection limit was 0.12 fg/,l and in TaqMan PCR 1.2 fg/,l, and real-time PCR was 104,105 times more sensitive than conventional PCR. The simple and rapid procedures maximized the yield and quality of recovered DNA from soil and allowed the processing of many samples in a short time. The direct DNA extractions from soil were utilized to yield DNA suitable for PCR. By combining this protocol with the real-time PCR procedure it has been possible to specifically detect P. nicotianae in soil, and the degree of sensitivity was 1.0 pg/,l. The system was applied to survey soil samples from tobacco field sites in China for the presence of P. nicotianae and the analyses of naturally infested soil showed the reliability of the real-time PCR method. [source] Screening of Plant Extracts for Antioxidant Activity: a Comparative Study on Three Testing MethodsPHYTOCHEMICAL ANALYSIS, Issue 1 2002Irina I. Koleva Abstract Three methods widely employed in the evaluation of antioxidant activity, namely 2,2,-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging method, static headspace gas chromatography (HS-GC) and ,-carotene bleaching test (BCBT), have been compared with regard to their application in the screening of plant extracts. The strengths and limitations of each method have been illustrated by testing a number of extracts, of differing polarity, from plants of the genus Sideritis, and two known antioxidants (butylated hydroxytoluene and rosmarinic acid). The sample polarity was important for the exhibited activity in the BCBT and HS-GC methods but not for the DPPH method. The complex composition of the extracts and partition phenomena affected their activity in each assay. The value of the BCBT method appears to be limited to less polar samples. Although slow, the HS-GC method is preferable for assessing the antioxidant inhibitory properties on the formation of unwanted secondary volatile products. Being rapid, simple and independent of sample polarity, the DPPH method is very convenient for the quick screening of many samples for radical scavenging activity. Copyright © 2001 John Wiley & Sons, Ltd. [source] A rapid and inexpensive microplate assay for the enzymatic determination of glucose, fructose, sucrose, L -malate and citrate in tomato (Lycopersicon esculentum) extracts and in orange juicePHYTOCHEMICAL ANALYSIS, Issue 5 2001Joyce S. Velterop Abstract Sugars and organic acids are important determinants of taste in fruits and vegetables. Based on the enzymatic analyses originally developed by Boehringer-Mannheim, rapid and reliable microplate-based assays were devised for glucose, fructose and sucrose in tomato and orange juice, and citrate and L -malate in tomato. These microplate assays were used to compare some commercially available tomato types, and to study the compartmentation of sugars and acids between pericarp and locular tissue of tomato fruit. The microplate format allows analysis of many samples per day and the only apparatus required is a microplate reader. Copyright © 2001 John Wiley & Sons, Ltd. [source] Faster and easier chromatin immunoprecipitation assay with high sensitivityPROTEINS: STRUCTURE, FUNCTION AND BIOINFORMATICS, Issue 1 2007Hidetsugu Kohzaki Dr. Abstract Chromatin immunoprecipitation (ChIP) assays are widely used to investigate where chromatin-binding proteins bind to the genome. The standard assay is very time consuming. We have developed a rapid ChIP assay in which the immunoprecipitates serve directly as PCR templates. This assay eliminates the step to reverse the crosslinking, shortening the assay by 1,day. It also requires a less immunoprecipitating antibody, permits many samples to be tested simultaneously, and is more sensitive than the standard ChIP assay. [source] Mineralogical And Chemical Investigations Of Bloomery Slags From Prehistoric (8th Century Bc To 4th Century Ad) Iron Production Sites In Upper And Lower Lusatia, GermanyARCHAEOMETRY, Issue 2 2001R. B. Heimann More than 400 fayalitic bloomery slags from prehistoric iron production sites in Upper and Lower Lusatia, eastern Germany, as well as bog iron ore samples and intermediary samples of the smelting process, were analysed by chemical and mineralogical techniques. While the precursor bog iron ores exploited in the two regions under investigation were very similar in composition, consisting of low-manganese/low-barium as well as high-manganese/high-barium types of ore, pronounced differences in slag composition were detected. Slags from 17 investigated sites in Upper Lusatia showed average P2O5 contents between 1 and 3 mass%, whereas slags from 15 investigated sites in Lower Lusatia were generally much richer in phosphorus, reaching values as high as 7 mass% P2O5. Since a reasonable correlation exists between calcium and phosphorus contents in the slags of the latter sites, it is conjectured that deliberate addition of CaO to the ore/charcoal charge of the bloomery furnace may have taken place in order to fix the phosphorus in the slags effectively. In many samples, this conjecture is being supported by the detection of a slag mineral Ca,Fe phosphate Ca9,xFe1+x(PO4)7 that presumably crystallized from a residual phosphorus-rich melt and shows a cotectic relationship to both Ca-rich fayalite and wustite, as well as to members of the solid solution series magnetite,hercynite. [source] Diffraction cartography: applying microbeams to macromolecular crystallography sample evaluation and data collectionACTA CRYSTALLOGRAPHICA SECTION D, Issue 8 2010Matthew W. Bowler Crystals of biological macromolecules often exhibit considerable inter-crystal and intra-crystal variation in diffraction quality. This requires the evaluation of many samples prior to data collection, a practice that is already widespread in macromolecular crystallography. As structural biologists move towards tackling ever more ambitious projects, new automated methods of sample evaluation will become crucial to the success of many projects, as will the availability of synchrotron-based facilities optimized for high-throughput evaluation of the diffraction characteristics of samples. Here, two examples of the types of advanced sample evaluation that will be required are presented: searching within a sample-containing loop for microcrystals using an X-ray beam of 5,µm diameter and selecting the most ordered regions of relatively large crystals using X-ray beams of 5,50,µm in diameter. A graphical user interface developed to assist with these screening methods is also presented. For the case in which the diffraction quality of a relatively large crystal is probed using a microbeam, the usefulness and implications of mapping diffraction-quality heterogeneity (diffraction cartography) are discussed. The implementation of these techniques in the context of planned upgrades to the ESRF's structural biology beamlines is also presented. [source] |