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Sample records for indirect immunofluorescent detection

  1. Indirect micro-immunofluorescence test for detecting type-specific antibodies to herpes simplex virus.

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    Forsey, T; Darougar, S

    1980-02-01

    A rapid indirect micro-immunofluorescence test capable of detecting and differentiating type-specific antibodies to herpes simplex virus is described. The test proved highly sensitive and, in 80 patients with active herpes ocular infection, antibody was detected in 94%. No anti-herpes antibody was detected in a control group of 20 patients with adenovirus infections. Testing of animal sera prepared against herpes simplex virus types 1 and 2 and of human sera from cases of ocular and genital herpes infections showed that the test can differentiate antibodies to the infecting serotypes. Specimens of whole blood, taken by fingerprick, and eye secretions, both collected on cellulose sponges, could be tested by indirect micro-immunofluorescence. Anti-herpes IgG, IgM, and IgA can also be detected.

  2. Human cryptosporidiosis: detection of specific antibodies in the serum by an indirect immunofluorescence

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    Braz Lúcia M.A.

    1996-01-01

    Full Text Available Cryptosporidium sp., a coccidian parasite usually found in the faeces of cattle, has been recently implicated as an agent of human intestinal disease, mainly in immunocompromised patients. In the study realized, by an indirect immunofluorescence technique, specific immunoglobulins (IgG and IgM have been demonstrated in human serum against Cryptosporidium oocysts. Purified oocysts were used as antigens in the indirect immunofluorecence assay. After analyzing this test in sera from selected groups of patients, the frequency of both specific IgG and IgM of immunocompetent children who were excreting oocysts in their faeces was 62% and in children with negative excretion of oocysts was 20% and 40%, respectively. In adults infected with the human immunodeficiency virus (HIV and who were excreting Cryptosporidium in their stools, the frequency was 57% for IgG but only 2% for IgM. Twenty three percent of immunocompromised adults with not determined excretion of oocysts in their stools had anti-Cryptosporidium IgG in their sera. Children infected with human immunodeficiency virus had no IgM and only 14% had IgG detectable in their sera. The indirect immunoflorescence assay, when used with other parasitological techniques appears to be useful for retrospective population studies and for diagnosis of acute infection. The humoral immune response of HIV positive patients to this protozoan agent needs clarification.

  3. Application of photostable quantum dots for indirect immunofluorescent detection of specific bacterial serotypes on small marine animals

    International Nuclear Information System (INIS)

    Decho, Alan W; Beckman, Erin M; Chandler, G Thomas; Kawaguchi, Tomohiro

    2008-01-01

    An indirect immunofluorescence approach was developed using semiconductor quantum dot nanocrystals to label and detect a specific bacterial serotype of the bacterial human pathogen Vibrio parahaemolyticus, attached to small marine animals (i.e. benthic harpacticoid copepods), which are suspected pathogen carriers. This photostable labeling method using nanotechnology will potentially allow specific serotypes of other bacterial pathogens to be detected with high sensitivity in a range of systems, and can be easily applied for sensitive detection to other Vibrio species such as Vibrio cholerae

  4. An Automatic Indirect Immunofluorescence Cell Segmentation System

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    Yung-Kuan Chan

    2014-01-01

    Full Text Available Indirect immunofluorescence (IIF with HEp-2 cells has been used for the detection of antinuclear autoantibodies (ANA in systemic autoimmune diseases. The ANA testing allows us to scan a broad range of autoantibody entities and to describe them by distinct fluorescence patterns. Automatic inspection for fluorescence patterns in an IIF image can assist physicians, without relevant experience, in making correct diagnosis. How to segment the cells from an IIF image is essential in developing an automatic inspection system for ANA testing. This paper focuses on the cell detection and segmentation; an efficient method is proposed for automatically detecting the cells with fluorescence pattern in an IIF image. Cell culture is a process in which cells grow under control. Cell counting technology plays an important role in measuring the cell density in a culture tank. Moreover, assessing medium suitability, determining population doubling times, and monitoring cell growth in cultures all require a means of quantifying cell population. The proposed method also can be used to count the cells from an image taken under a fluorescence microscope.

  5. Indirect immunofluorescence assay for the simultaneous detection of antibodies against clinically important old and new world hantaviruses.

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    Sabine Lederer

    Full Text Available In order to detect serum antibodies against clinically important Old and New World hantaviruses simultaneously, multiparametric indirect immunofluorescence assays (IFAs based on biochip mosaics were developed. Each of the mosaic substrates consisted of cells infected with one of the virus types Hantaan (HTNV, Puumala (PUUV, Seoul (SEOV, Saaremaa (SAAV, Dobrava (DOBV, Sin Nombre (SNV or Andes (ANDV. For assay evaluation, serum IgG and IgM antibodies were analyzed using 184 laboratory-confirmed hantavirus-positive sera collected at six diagnostic centers from patients actively or previously infected with the following hantavirus serotypes: PUUV (Finland, n=97; SEOV (China, n=5; DOBV (Romania, n=7; SNV (Canada, n=23; ANDV (Argentina and Chile, n=52. The control panel comprised 89 sera from healthy blood donors. According to the reference tests, all 184 patient samples were seropositive for hantavirus-specific IgG (n=177; 96% and/or IgM (n=131; 72%, while all control samples were tested negative. In the multiparametric IFA applied in this study, 183 (99% of the patient sera were IgG and 131 (71% IgM positive (accordance with the reference tests: IgG, 96%; IgM, 93%. Overall IFA sensitivity for combined IgG and IgM analysis amounted to 100% for all serotypes, except for SNV (96%. Of the 89 control sera, 2 (2% showed IgG reactivity against the HTNV substrate, but not against any other hantavirus. Due to the high cross-reactivity of hantaviral nucleocapsid proteins, endpoint titrations were conducted, allowing serotype determination in >90% of PUUV- and ANDV-infected patients. Thus, multiparametric IFA enables highly sensitive and specific serological diagnosis of hantavirus infections and can be used to differentiate PUUV and ANDV infection from infections with Murinae-borne hantaviruses (e.g. DOBV and SEOV.

  6. Comparison of electron microscopy, enzyme-linked immunosorbent assay, solid-phase radioimmunoassay, and indirect immunofluorescence for detection of human rotavirus antigen in faeces

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    Birch, C J; Lehmann, N I; Hawker, A J; Marshall, J A; Gust, I D [Fairfield Hospital for Communicable Diseases, Victoria (Australia). Virology Dept.

    1979-07-01

    Four techniques were compared for their practicability, speed, and sensitivity for the detection of human rotavirus. Radioimmunoassay (RIA) and enzyme-linked immunosorbent assay (ELISA) were found to be the most sensitive means of identifying rotavirus and, once processed, up to 40 specimens could be examined daily. Electron microscopy, although less sensitive than these techniques, had the advantage of being able to detect other viral agents present in faecal extracts. Indirect immunofluorescence failed to detect rotavirus as often as the other three methods. In laboratories where routine examination of faecal specimens from patients with gastroenteritis is required, ELISA and RIA are useful alternatives to electron microscopy.

  7. Comparison of electron microscopy, enzyme-linked immunosorbent assay, solid-phase radioimmunoassay, and indirect immunofluorescence for detection of human rotavirus antigen in faeces

    International Nuclear Information System (INIS)

    Birch, C.J.; Lehmann, N.I.; Hawker, A.J.; Marshall, J.A.; Gust, I.D.

    1979-01-01

    Four techniques were compared for their practicability, speed, and sensitivity for the detection of human rotavirus. Radioimmunoassay (RIA) and enzyme-linked immunosorbent assay (ELISA) were found to be the most sensitive means of identifying rotavirus and, once processed, up to 40 specimens could be examined daily. Electron microscopy, although less sensitive than these techniques, had the advantage of being able to detect other viral agents present in faecal extracts. Indirect immunofluorescence failed to detect rotavirus as often as the other three methods. In laboratories where routine examination of faecal specimens from patients with gastroenteritis is required, ELISA and RIA are useful alternatives to electron microscopy. (author)

  8. [Standardized indirect immunofluorescence. Differentiation of mitochondrial, microsomal and ribosomal antibodies].

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    Storch, W

    1977-02-15

    By an extensive standardisation of the indirect immunofluorescence for the demonstration espeially of mitochondrial antibodies we succeeded in recognizing atypical fluorescence patterns and in describing their exact localisation. On the basis of absorption studies with mitochondrias, microsomas and ribosomas by comparative observation of sections of liver, stomach and kidneys of rats the preferred sort of reaction and the intensity of fluorescence of antibodies against mitochondria, microsomas and ribosomas were empirically established. Antimitochondrial antibodies react above all with the parietal cells of the stomach and the distal epithelia of the tubulus of the kidney. Antibodies against microsomas of liver and kidney are characterized by a brilliant diffuse cytoplasmatic fluorescence of the hepatocytes and by a comparatively weaker fluorescence of exclusively proximal tubuli of the kidneys of rats. Antibodies against ribosomas lead to a fluorescence especially of the main cells of the stomach. The differentiation of several cytoplasmatic antibodies is among others of interest for the diagnosis of certain autoimmune diseases. Although there are numerous still unclear findings and "overlap" phenomena the existence of high titre antibodies against mitochondrias speaks for a primarily biliary cirrhosis or a pseudo-LE-syndrome, the existence of antibodies against microsomas of kidney and liver of rats for a special form of a chronically active hepatitis and the existence of the very rare antibodies against ribosomas for an active lupus erythematodes disseminatus.

  9. Immunofluorescence

    International Nuclear Information System (INIS)

    Bongertz, V.; Castro, B.G.

    1990-01-01

    The advantages of the immunofluorescence assay (IFA) that allowing the specific 'in situ' localization of the antigen-antibody interaction, of paramount importance when complex antigenic preparations, such as for instance tissue sections, are studied. (L.M.J.) [pt

  10. Indirect immunofluorescence staining of Chlamydia trachomatis inclusions in microculture plates with monoclonal antibodies.

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    Zapata, M; Chernesky, M; Mahony, J

    1984-06-01

    Indirect immunofluorescence (IF) staining, using a monoclonal antibody, detected two- to fourfold more inclusions than did iodine staining. Of 274 clinical specimens, 53 (19.3%) were positive by IF on passage 1 as compared with 33 (12%) by iodine staining (P less than 0.005). IF-stained inclusions in McCoy cells in the bottom of microculture wells were readily viewed with a long-focal-length objective at a magnification of 250 X.

  11. Indirect immunofluorescence staining of Chlamydia trachomatis inclusions in microculture plates with monoclonal antibodies.

    OpenAIRE

    Zapata, M; Chernesky, M; Mahony, J

    1984-01-01

    Indirect immunofluorescence (IF) staining, using a monoclonal antibody, detected two- to fourfold more inclusions than did iodine staining. Of 274 clinical specimens, 53 (19.3%) were positive by IF on passage 1 as compared with 33 (12%) by iodine staining (P less than 0.005). IF-stained inclusions in McCoy cells in the bottom of microculture wells were readily viewed with a long-focal-length objective at a magnification of 250 X.

  12. Epidermis area detection for immunofluorescence microscopy

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    Dovganich, Andrey; Krylov, Andrey; Nasonov, Andrey; Makhneva, Natalia

    2018-04-01

    We propose a novel image segmentation method for immunofluorescence microscopy images of skin tissue for the diagnosis of various skin diseases. The segmentation is based on machine learning algorithms. The feature vector is filled by three groups of features: statistical features, Laws' texture energy measures and local binary patterns. The images are preprocessed for better learning. Different machine learning algorithms have been used and the best results have been obtained with random forest algorithm. We use the proposed method to detect the epidermis region as a part of pemphigus diagnosis system.

  13. Detection of Streptococcus suis by in situ hybridization, indirect immunofluorescence, and peroxidase-antiperoxidase assays in formalin-fixed, paraffin-embedded tissue sections from pigs

    DEFF Research Database (Denmark)

    Boye, Mette; Feenstra, Anne Avlund; Tegtmeier, Conny

    2000-01-01

    and the immunohistochemical methods were used for detection of S. suis in formalin-fixed, paraffin-embedded tissue sections of brain, endocardium, and lung from pigs infected with S. suis. The methods developed were able to detect single cells of S. suis in situ in the respective samples, whereas no signal was observed from...

  14. Original Approach for Automated Quantification of Antinuclear Autoantibodies by Indirect Immunofluorescence

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    Daniel Bertin

    2013-01-01

    Full Text Available Introduction. Indirect immunofluorescence (IIF is the gold standard method for the detection of antinuclear antibodies (ANA which are essential markers for the diagnosis of systemic autoimmune rheumatic diseases. For the discrimination of positive and negative samples, we propose here an original approach named Immunofluorescence for Computed Antinuclear antibody Rational Evaluation (ICARE based on the calculation of a fluorescence index (FI. Methods. We made comparison between FI and visual evaluations on 237 consecutive samples and on a cohort of 25 patients with SLE. Results. We obtained very good technical performance of FI (95% sensitivity, 98% specificity, and a kappa of 0.92, even in a subgroup of weakly positive samples. A significant correlation between quantification of FI and IIF ANA titers was found (Spearman's ρ=0.80, P<0.0001. Clinical performance of ICARE was validated on a cohort of patients with SLE corroborating the fact that FI could represent an attractive alternative for the evaluation of antibody titer. Conclusion. Our results represent a major step for automated quantification of IIF ANA, opening attractive perspectives such as rapid sample screening and laboratory standardization.

  15. Hepatitis B Core Antigen in Hepatocytes of Chronic Hepatitis B: Comparison between Indirect Immunofluorescence and Immunoperoxidase Method

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    Tabassum, Shahina; Al-Mahtab, Mamun; Nessa, Afzalun; Jahan, Munira; Shamim Kabir, Chowdhury Mohammad; Kamal, Mohammad; Cesar Aguilar, Julio

    2015-01-01

    Background Hepatitis B virus (HBV) infection has many faces. Precore and core promoter mutants resemble inactive carrier status. The identification of hepatitis B core antigen (HBcAg) in hepatocytes may have variable clinical significance. The present study was undertaken to detect HBcAg in chronic hepatitis B (CHB) patients and to assess the efficacy of detection system by indirect immunofluorescence (IIF) and indirect immunoperoxidase (IIP). Materials and methods The study was done in 70 chronic HBV-infected patients. Out of 70 patients, eight (11.4%) were hepatitis B e antigen (HBeAg) positive and 62 (88.57%) were HBeAg negative. Hepatitis B core antigen was detected by indirect immunofluorescence (IIF) and indirect immunoperoxidase (IIP) methods in liver tissue. Results All HBeAg positive patients expressed HBcAg by both IIF and IIP methods. Out of 62 patients with HBeAg-negative CHB, HBcAg was detected by IIF in 55 (88.7%) patients and by IIP in 51 (82.26%) patients. A positive relation among viral load and HBcAg detection was also found. This was more evident in the case of HBeAg negative patients and showed a positive relation with HBV DNA levels. Conclusion Hepatitis B core antigen can be detected using the IIF from formalin fixed paraffin block preparation and also by IIP method. This seems to reflect the magnitudes of HBV replication in CHB. How to cite this article Raihan R, Tabassum S, Al-Mahtab M, Nessa A, Jahan M, Kabir CMS, Kamal M, Aguilar JC. Hepatitis B Core Antigen in Hepatocytes of Chronic Hepatitis B: Comparison between Indirect Immunofluorescence and Immunoperoxidase Method. Euroasian J Hepato-Gastroenterol 2015;5(1):7-10. PMID:29201677

  16. Antibodies to UV irradiated DNA: the monitoring of DNA damage by ELISA and indirect immunofluorescence

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    Wani, A A; Gibson-D' Ambrosio, R E; D' Ambrosio, S M [Ohio State Univ., Columbus (USA). Dept. of Radiology

    1984-10-01

    The enzyme-linked immunosorbant assay (ELISA) was modified to (1) characterize antibodies raised in rabbits against UV-irradiated single-stranded DNA (UVssDNA) complexed with methylated BSA and (2) directly detect pyrimidine dimers in irradiated DNA. The antisera specifically bound to UVssDNA, UVpoly(dT) and to a limited extent to UVdsDNA and UVpoly(dC). Fifty per cent of the maximum antibody binding was observed at a 1-5000 dilution against UVssDNA. Binding to ssDNA and poly(dT) was observed only at much higher concentrations of antibody, whereas no binding to double stranded DNA (dsDNA) was observed. The extent of binding of the antibody was dependent on the UV dose to DNA and the concentration of antigen immobilized on the plate. The ability of various irradiated molecules, DNA, homopolymers and linkers to act as inhibitors of antibody binding establishes that the antigenic determinants are mainly thymine homodimers with lower affinity for cytosine dimers. Potential usefulness of the antibodies to directly quantitate pyrimidine dimers in cells exposed to UV radiation was determined by indirect immunofluorescence. Flow cytometric analysis of immunostained human lymphocytes irradiated with 254 nm radiation indicated that greater than 50% of the population had significantly higher fluorescent intensity than unirradiated cells.

  17. Immunofluorescence detection of pea protein in meat products.

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    Petrášová, Michaela; Pospiech, Matej; Tremlová, Bohuslava; Javůrková, Zdeňka

    2016-08-01

    In this study we developed an immunofluorescence method to detect pea protein in meat products. Pea protein has a high nutritional value but in sensitive individuals it may be responsible for causing allergic reactions. We produced model meat products with various additions of pea protein and flour; the detection limit (LOD) of the method for pea flour was 0.5% addition, and for pea protein it was 0.001% addition. The repeatabilities and reproducibilities for samples both positive and negative for pea protein were all 100%. In a blind test with model products and commercial samples, there was no statistically significant difference (p > 0.05) between the declared concentrations of pea protein and flour and the immunofluorescence method results. Sensitivity was 1.06 and specificity was 1.00. These results show that the immunofluorescence method is suitable for the detection of pea protein in meat products.

  18. Improved detection of Pneumocystis carinii by an immunofluorescence technique using monoclonal antibodies

    DEFF Research Database (Denmark)

    Orholm, M; Holten-Andersen, W; Lundgren, Jens Dilling

    1990-01-01

    To assess whether a recently developed indirect immunofluorescent stain using monoclonal antibodies was more sensitive in detecting Pneumocystis carinii than the combination of Giemsa and methenamine silver nitrate stains which has routinely been used in the laboratory, 88 lavage fluid specimens...... silver nitrate and toluidine blue O. Immunofluorescence using the monoclonal antibodies from the NIH was significantly more sensitive than any other single staining method and than the combination of Giemsa and methenamine silver nitrate staining. The study also showed that the cytospin centrifuge...

  19. Application of indirect immunofluorescent test with an improved HEp-2 substrate tranfected with human Ro60/SSA autoantigens

    International Nuclear Information System (INIS)

    Lv Liangjing; Chen Shunle; Gu Yueying; Shen Nan; Bao Chunde; Wang Yuan; Xue Feng; Ye Peng; Yu Chongzhao

    2006-01-01

    To develop an improved substrate for indirect immunofluorescent test (IIF) to detect anti-Ro/SSA autoantibodies, the human 60-kDa Ro/SSA autoantigens (Ro60) cDNAs were obtained from placental cDNA library using PCR and were cloned into the mammalian expression vectorpEGFP-C1. Then the recombinant plasmids were transfected into HEp-2 cells. We con- firmed the overexpression, localization and antigenicity of fusion proteins in transfected cells by means of fluorescence microscopy, immunoblotting and IIF. HEp-2 and HEp-Ro60 was analyzed by IIF using a panel of 10 precipitinpositive anti-Ro human sera simultaneously. Stable expression of Ro60-GFP (green fluorescent protein) fusion proteins maintained ten more generations. And Ro60-GFP kept the antigenicity of Ro and had its own characteristic immunofluorescent pattern in HEp-Ro60 cells. The transfectants dramatically increased the sensitivity of IIF testing (a mean increase of 6.7-fold in endpoint titer, P<0.01). Eight (8/10) positive an- ti-Ro sera showed characteristic immunofluorescent pattern on HEp-Ro60, including two sera which were antinuclear antibodies (ANA) negative on untransfected HEp-2. IIF-ANA in all healthy sera were negative on HEp-Ro60. As a kind of new substrate of IIF, the Ro60 transfectants can be used to detect anti-Ro antibodies. In addition, transfected HEp-2 cells kept the immunofluorescent property of HEp-2 cells in IIF-ANA tests and could be employed as substrate for the routine IIF-ANA detection. The method improved the sensitivity of IIF-ANA. (authors)

  20. A comparison of indirect immunofluorescence and electron microscopy for the diagnosis of some haemorrhagic viruses in cell cultures.

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    El Mekki, A A; van der Groen, G

    1981-09-01

    Yellow fever, dengue (types 1, 2 and 4), Chikungunya, Rift Valley fever, Ebola, Marburg, and Lassa viruses were inoculated into susceptible cell cultures and daily investigated by indirect immunofluorescence (IFA) and electron microscopy (EM) with a view to achieve an early detection-identification of these agents. Compared to the other cell lines tested (Vero, BHK-21 and Aedes albopictus), CV-1 cells were found to be more sensitive. Viral antigens were detected by IFA from a few hours post inoculation (CHIK and RVF) to a maximum of 3 days (YF and EBO). For most of the viruses studied, the cytopathic effect (CPE) commenced 2-3 days after the detection of viral antigens. Virus particles were detected by EM only in the case of EBO, MBG and LAS, before any CPE was observed in cell cultures.

  1. Staining pattern classification of antinuclear autoantibodies based on block segmentation in indirect immunofluorescence images.

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    Jiaqian Li

    Full Text Available Indirect immunofluorescence based on HEp-2 cell substrate is the most commonly used staining method for antinuclear autoantibodies associated with different types of autoimmune pathologies. The aim of this paper is to design an automatic system to identify the staining patterns based on block segmentation compared to the cell segmentation most used in previous research. Various feature descriptors and classifiers are tested and compared in the classification of the staining pattern of blocks and it is found that the technique of the combination of the local binary pattern and the k-nearest neighbor algorithm achieve the best performance. Relying on the results of block pattern classification, experiments on the whole images show that classifier fusion rules are able to identify the staining patterns of the whole well (specimen image with a total accuracy of about 94.62%.

  2. Automated Processing and Evaluation of Anti-Nuclear Antibody Indirect Immunofluorescence Testing.

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    Ricchiuti, Vincent; Adams, Joseph; Hardy, Donna J; Katayev, Alexander; Fleming, James K

    2018-01-01

    Indirect immunofluorescence (IIF) is considered by the American College of Rheumatology (ACR) and the international consensus on ANA patterns (ICAP) the gold standard for the screening of anti-nuclear antibodies (ANA). As conventional IIF is labor intensive, time-consuming, subjective, and poorly standardized, there have been ongoing efforts to improve the standardization of reagents and to develop automated platforms for assay incubation, microscopy, and evaluation. In this study, the workflow and performance characteristics of a fully automated ANA IIF system (Sprinter XL, EUROPattern Suite, IFA 40: HEp-20-10 cells) were compared to a manual approach using visual microscopy with a filter device for single-well titration and to technologist reading. The Sprinter/EUROPattern system enabled the processing of large daily workload cohorts in less than 8 h and the reduction of labor hands-on time by more than 4 h. Regarding the discrimination of positive from negative samples, the overall agreement of the EUROPattern software with technologist reading was higher (95.6%) than when compared to the current method (89.4%). Moreover, the software was consistent with technologist reading in 80.6-97.5% of patterns and 71.0-93.8% of titers. In conclusion, the Sprinter/EUROPattern system provides substantial labor savings and good concordance with technologist ANA IIF microscopy, thus increasing standardization, laboratory efficiency, and removing subjectivity.

  3. Automated Processing and Evaluation of Anti-Nuclear Antibody Indirect Immunofluorescence Testing

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    Vincent Ricchiuti

    2018-05-01

    Full Text Available Indirect immunofluorescence (IIF is considered by the American College of Rheumatology (ACR and the international consensus on ANA patterns (ICAP the gold standard for the screening of anti-nuclear antibodies (ANA. As conventional IIF is labor intensive, time-consuming, subjective, and poorly standardized, there have been ongoing efforts to improve the standardization of reagents and to develop automated platforms for assay incubation, microscopy, and evaluation. In this study, the workflow and performance characteristics of a fully automated ANA IIF system (Sprinter XL, EUROPattern Suite, IFA 40: HEp-20-10 cells were compared to a manual approach using visual microscopy with a filter device for single-well titration and to technologist reading. The Sprinter/EUROPattern system enabled the processing of large daily workload cohorts in less than 8 h and the reduction of labor hands-on time by more than 4 h. Regarding the discrimination of positive from negative samples, the overall agreement of the EUROPattern software with technologist reading was higher (95.6% than when compared to the current method (89.4%. Moreover, the software was consistent with technologist reading in 80.6–97.5% of patterns and 71.0–93.8% of titers. In conclusion, the Sprinter/EUROPattern system provides substantial labor savings and good concordance with technologist ANA IIF microscopy, thus increasing standardization, laboratory efficiency, and removing subjectivity.

  4. Performance of an ELISA and Indirect Immunofluorescence Assay in Serological Diagnosis of Zoonotic Cutaneous Leishmaniasis in Iran

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    Bahador Sarkari

    2014-01-01

    Full Text Available Serological assays have been extensively evaluated for diagnosis of visceral leishmaniasis (VL and considered as a routine method for diagnosis of VL while these methods are not properly evaluated for diagnosis of cutaneous leishmaniasis (CL. This study aimed to assess the performance of indirect immunofluorescent-antibody test (IFA and enzyme-linked immunosorbent assay (ELISA for serodiagnosis of cutaneous leishmaniasis in Iran. Sixty-one sera samples from parasitologically confirmed CL patients and 50 sera from healthy controls along with 50 sera from non-CL patients were collected. Antigen was prepared from promastigotes and amastigotes of Leishmania major. IFA was used to detect anti-Leishmania IgG while ELISA was used to detect anti-Leishmania IgM, total IgG, or IgG subclasses (IgG1 and 4. ELISA, for detection of total IgG and IgM, showed sensitivity of 83.6% and 84.7% and specificity of 62.7% and 54.6%, respectively. Sensitivity and specificity of ELISA for detecting IgG1 and IgG4 were 64%, 75% and 85%, 49%, respectively. Sensitivity and specificity of IFA were 91.6% and 81%. Conclusion. Findings of this study demonstrated that serological test, especially IFA, can be used for proper diagnosis of CL.

  5. Immunofluorescence detection of milk protein in meat products

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    Michaela Petrášová

    2015-05-01

    Full Text Available Nowadays there are various vegetable protein additives intended for the manufacture of meat products in the food industry. These ingredients include both, plant-origin as well as animal-origin proteins. The most common vegetable additives include various types of flour, starch, fiber and plant protein. Among animal proteins, the most commonly used are plasma, collagen or milk protein. Milk protein is added to meat products due to its functional properties, such as emulsifying fats, improving the holding capacity of meat, improving juiciness, gel-forming capacity and affecting the taste of the product. Usage of these proteins, however, is currently limited by the effective legislation, not only in order to prevent consumer deception, but also because of their potential impact on consumers' health of. Thus, this issue has received considerable attention not only in the Czech Republic, but also globally. The main risk is the impossibility of selecting a suitable foodstuff for individuals with potential allergic reactions. The only option for allergic consumers to protect themselves is to strictly exclude the given allergen from their diet. Although the number of studies dealing with the reduction or loss of allergenicity is increasing, yet these practices are not common. Most of the population suffering from food allergies is thus still dependent on strict exclusion of foodstuffs causing adverse allergic reactions from their diet. Detection of allergens in foodstuffs is unfortunately quite difficult due to the fact that they occur in trace amounts and are often masked by different parts of the foodstuff. This research dealt with the detection of milk protein in meat products purchased in the market network of the Czech Republic, whereas declaration given by the manufacturer on the packaging for the small meat products purchased from the market was used to verify the detection of milk protein by the immunofluorescence method. 20 products were

  6. Supersymmetric dark matter: Indirect detection

    International Nuclear Information System (INIS)

    Bergstroem, L.

    2000-01-01

    Dark matter detection experiments are improving to the point where they can detect or restrict the primary particle physics candidates for non baryonic dark matter. The methods for detection are usually categorized as direct, i.e., searching for signals caused by passage of dark matter particles in terrestrial detectors, or indirect. Indirect detection methods include searching for antimatter and gamma rays, in particular gamma ray lines, in cosmic rays and high-energy neutrinos from the centre of the Earth or Sun caused by accretion and annihilation of dark matter particles. A review is given of recent progress in indirect detection, both on the theoretical and experimental side

  7. Detection of serum antitrichomonal antibodies in urogenital trichomoniasis by immunofluorescence.

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    Bhatt R

    1992-04-01

    Full Text Available Trichomonas vaginalis is a frequently encountered genital pathogen in both males and females. In females, vaginitis due to this parasite is one of the most common manifestation. The indirect fluorescent technique (IFA test was carried out to detect antitrichomonal antibodies in 370 female patients using whole cell antigen. Seventy one (19.18% gave positive reaction for either of the class IgG, IgM and IgA antibodies. The level of the IgG class antibodies was found to be higher i.e. 58 (81.69% than IgM 11 (15.27% antibodies, which may be suggestive of past infection or a prolonged manifestation by the organisms.

  8. Performance analysis of automated evaluation of Crithidia luciliae-based indirect immunofluorescence tests in a routine setting - strengths and weaknesses.

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    Hormann, Wymke; Hahn, Melanie; Gerlach, Stefan; Hochstrate, Nicola; Affeldt, Kai; Giesen, Joyce; Fechner, Kai; Damoiseaux, Jan G M C

    2017-11-27

    Antibodies directed against dsDNA are a highly specific diagnostic marker for the presence of systemic lupus erythematosus and of particular importance in its diagnosis. To assess anti-dsDNA antibodies, the Crithidia luciliae-based indirect immunofluorescence test (CLIFT) is one of the assays considered to be the best choice. To overcome the drawback of subjective result interpretation that inheres indirect immunofluorescence assays in general, automated systems have been introduced into the market during the last years. Among these systems is the EUROPattern Suite, an advanced automated fluorescence microscope equipped with different software packages, capable of automated pattern interpretation and result suggestion for ANA, ANCA and CLIFT analysis. We analyzed the performance of the EUROPattern Suite with its automated fluorescence interpretation for CLIFT in a routine setting, reflecting the everyday life of a diagnostic laboratory. Three hundred and twelve consecutive samples were collected, sent to the Central Diagnostic Laboratory of the Maastricht University Medical Centre with a request for anti-dsDNA analysis over a period of 7 months. Agreement between EUROPattern assay analysis and the visual read was 93.3%. Sensitivity and specificity were 94.1% and 93.2%, respectively. The EUROPattern Suite performed reliably and greatly supported result interpretation. Automated image acquisition is readily performed and automated image classification gives a reliable recommendation for assay evaluation to the operator. The EUROPattern Suite optimizes workflow and contributes to standardization between different operators or laboratories.

  9. Aquaporin-4 autoantibodies in neuromyelitis optica spectrum disorders: comparison between tissue-based and cell-based indirect immunofluorescence assays

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    Chan Koon H

    2010-09-01

    Full Text Available Abstract Background Neuromyelitis optica spectrum disorders (NMOSD are severe central nervous system inflammatory demyelinating disorders (CNS IDD characterized by monophasic or relapsing, longitudinally extensive transverse myelitis (LETM and/or optic neuritis (ON. A significant proportion of NMOSD patients are seropositive for aquaporin-4 (AQP4 autoantibodies. We compared the AQP4 autoantibody detection rates of tissue-based indirect immunofluorescence assay (IIFA and cell-based IIFA. Methods Serum of Chinese CNS IDD patients were assayed for AQP4 autoantibodies by tissue-based IIFA using monkey cerebellum and cell-based IIFA using transfected HEK293 cells which express human AQP4 on their cell membranes. Results In total, 128 CNS IDD patients were studied. We found that 78% of NMO patients were seropositive for AQP4 autoantibodies by cell-based IIFA versus 61% by tissue-based IFA (p = 0.250, 75% of patients having relapsing myelitis (RM with LETM were seropositive by cell-based IIFA versus 50% by tissue-based IIFA (p = 0.250, and 33% of relapsing ON patients were seropositive by cell-based IIFA versus 22% by tissue-based IIFA (p = 1.000; however the differences were not statistically significant. All patients seropositive by tissue-based IIFA were also seropositive for AQP4 autoantibodies by cell-based IIFA. Among 29 NMOSD patients seropositive for AQP4 autoantibodies by cell-based IIFA, 20 (69% were seropositive by tissue-based IIFA. The 9 patients seropositive by cell-based IIFA while seronegative by tissue-based IIFA had NMO (3, RM with LETM (3, a single attack of LETM (1, relapsing ON (1 and a single ON attack (1. Among 23 NMO or RM patients seropositive for AQP4 autoantibodies by cell-based IIFA, comparison between those seropositive (n = 17 and seronegative (n = 6 by tissue-based IIFA revealed no differences in clinical and neuroradiological characteristics between the two groups. Conclusion Cell-based IIFA is slightly more sensitive

  10. Accuracy of molecular diagnostics in pemphigus and bullous pemphigoid: comparison of commercial and modified mosaic indirect immunofluorescence tests as well as enzyme-linked immunosorbent assays.

    Science.gov (United States)

    Gornowicz-Porowska, Justyna; Seraszek-Jaros, Agnieszka; Bowszyc-Dmochowska, Monika; Kaczmarek, Elżbieta; Pietkiewicz, Paweł; Bartkiewicz, Paweł; Dmochowski, Marian

    2017-02-01

    Pemphigus and bullous pemphigoid (BP) are identified by autoantibodies (abs) against desmoglein 1, 3 (DSG1/3) and BP180/BP230, respectively. A novel mosaic to indirect immunofluorescence (IIF) using purified BP180 recombinant proteins spotted on slide and transfected cells expressing BP230, DSG1, DSG3 is available. The commercial (IgG detection) and modified (IgG4 detection) mosaic for indirect immunofluorescence (IIFc - IIF commercial, IIFm - IIF modified) and IgG ELISAs were evaluated in pemphigus and bullous pemphigoid (BP) molecular diagnostics. To compare diagnostic accuracy of commercial (IgG detection) and modified (IgG4 detection) mosaic IIF assay and to examine the diagnostic value of ELISAs in relation to mosaic IIF in routine laboratory diagnostics of pemphigus and BP. Sera from 37 BP and 19 pemphigus patients were studied. Associations between tests were assessed using Fisher's exact test. There are associations between the positive/negative samples detected by IIFc with desmoglein1 (DSG1)/desmoglein3 (DSG3)/BP230 transfected cells and ELISAs and no association between anti-BP180 IgG detection by IIFc and ELISA. IIFm with DSG1 and DSG3 showed both 100% sensitivity and 100% and 78% specificity, respectively, and 100% and 83% positive predictive value in relation to IIFc. IIFm with BP230 had 87% specificity, 55% sensitivity, whereas IIFm with BP180 had a 100% sensitivity and 13% specificity in relation to IIFc. The IIFc with DSG1/DSG3/BP230 transfected cells, excluding BP180 spots, is an alternative method to ELISA in pemphigus/BP diagnostics. IgG4 antibodies, both pathogenically and diagnostically important, are inconsistently detectable with IIFm.

  11. Accuracy of molecular diagnostics in pemphigus and bullous pemphigoid: comparison of commercial and modified mosaic indirect immunofluorescence tests as well as enzyme-linked immunosorbent assays

    Directory of Open Access Journals (Sweden)

    Justyna Gornowicz-Porowska

    2017-02-01

    Full Text Available Introduction : Pemphigus and bullous pemphigoid (BP are identified by autoantibodies (abs against desmoglein 1, 3 (DSG1/3 and BP180/BP230, respectively. A novel mosaic to indirect immunofluorescence (IIF using purified BP180 recombinant proteins spotted on slide and transfected cells expressing BP230, DSG1, DSG3 is available. The commercial (IgG detection and modified (IgG4 detection mosaic for indirect immunofluorescence (IIFc – IIF commercial, IIFm – IIF modified and IgG ELISAs were evaluated in pemphigus and bullous pemphigoid (BP molecular diagnostics. Aim : To compare diagnostic accuracy of commercial (IgG detection and modified (IgG4 detection mosaic IIF assay and to examine the diagnostic value of ELISAs in relation to mosaic IIF in routine laboratory diagnostics of pemphigus and BP. Material and methods : Sera from 37 BP and 19 pemphigus patients were studied. Associations between tests were assessed using Fisher’s exact test. Results: There are associations between the positive/negative samples detected by IIFc with desmoglein1 (DSG1/desmoglein3 (DSG3/BP230 transfected cells and ELISAs and no association between anti-BP180 IgG detection by IIFc and ELISA. IIFm with DSG1 and DSG3 showed both 100% sensitivity and 100% and 78% specificity, respectively, and 100% and 83% positive predictive value in relation to IIFc. IIFm with BP230 had 87% specificity, 55% sensitivity, whereas IIFm with BP180 had a 100% sensitivity and 13% specificity in relation to IIFc. Conclusions : The IIFc with DSG1/DSG3/BP230 transfected cells, excluding BP180 spots, is an alternative method to ELISA in pemphigus/BP diagnostics. IgG4 antibodies, both pathogenically and diagnostically important, are inconsistently detectable with IIFm.

  12. Indirect detection of dark matter

    International Nuclear Information System (INIS)

    Pieri, L.

    2008-01-01

    In the Cold Dark Matter scenario, the Dark Matter particle candidate may be a Weakly Interacting Massive Particle (Wimp). Annihilation of two Wimps in local or cosmological structures would result in the production of a number of standard model particles such as photons, leptons and baryons which could be observed with the presently available or future experiments such as the Pamela or Glast satellites or the Cherenkov Telescopes. In this work we review the status-of-the-art of the theoretical and phenomenological studies about the possibility of indirect detection of signals coming from Wimp annihilation.

  13. HEMORRHAGIC-FEVER VIRUS-INFECTIONS IN AN ISOLATED RAIN-FOREST AREA OF CENTRAL LIBERIA - LIMITATIONS OF THE INDIRECT IMMUNOFLUORESCENCE SLIDE TEST FOR ANTIBODY SCREENING IN AFRICA

    NARCIS (Netherlands)

    van der Waals, F. W.; Pomeroy, K. L.; Goudsmit, J.; Asher, D. M.; Gajdusek, D. C.

    1986-01-01

    Serum samples from 119 healthy individuals and 106 epilepsy patients inhabiting Grand Bassa County, Liberia, were tested for antibodies to hemorrhagic fever viruses (HFV) by indirect immunofluorescence. E6 Vero cells infected with Lassa fever virus (LAS), Rift Valley Fever virus (RVF), Congo

  14. Longitudinal study of the indirect immunofluorescence and complement fixation tests for diagnosis of chagas' disease in immunosuppressed patients submitted to renal transplantation

    Directory of Open Access Journals (Sweden)

    José Fernando de Castro Figueiredo

    1993-12-01

    Full Text Available Clinical and serological follow-up of 7 patients submitted to renal transplantation and presenting positive serological reactions to Chagas 'disease before immunossupression did not show significant changes in indirect immunofluorescence and complement fixation titres for Chagas ' disease, or signs and symptoms indicating exacerbation of the disease during follow- up. In addition, 18 of 66 recipients of renal transplants considered to be non-chagasic before immunosuppression showed at least one positive result to the indirect immunofluorescence test for Chagas ' disease during the study period. The results suggest that the immunosuppression State induced in chagasic patients submitted to renal transplant did notpromoted exacerbation of the chronic infection in these patients and not interfere with the serological response of chronic chagasics, thus permitting the use of these serologic reactions for diagnostic purposes in these cases. However, the positive results ofthe indirect immunofluorescence test in non- chagasic patients indicate the needforjudicious interpretation ofthe indirect immunofluorescence test for the diagnosis of Chagas' disease in renal transplanted patients.

  15. Detection of viral infection by immunofluorescence in formalin-fixed tissues, pretreated with trypsin

    Directory of Open Access Journals (Sweden)

    O. M. Barth

    1988-06-01

    Full Text Available The presence of viral antigen in sections from formalin-fixed and paraffin-embedded human tissues was demonstrated by trypsin digestion followed by direct or indirect immunofluorescence. The specimens may be used for retrospective diagnosis. The immunofluorescence technique has to be adapted to the suspected virus infection on the basis of previous histopathology study. Variations of trypsin concentration time and temperature of incubation, expose different viral antigens and have to be previously tested for each unknown system. For measles virus detection in lung a stronger digestion has to be applied as compared to adenovirus or respiratory disease viruses in the same tisue. Flavivirus in liver tissue needs a weaker digestion. The reproducibility of the method makes it useful as a routine technique in diagnosis of virus infection.A presença de antígeno viral em cortes de tecidos humanos fixados em formol e emblocados em parafina foi demonstrada pela digestão com tripsina foi demonstrada pela ingestão com tripsina seguida de imunofluorescência direta ou indireta. Os espécimens podem ser utilizados para diagnoses retrospectivas. A técnica da imunofluorescência deve ser adaptada à infecção viral suspeita segundo diagnosie histopatológica prévia. Os parâmetros para a digestão do tecido pela tripsina, relacionados à concentração, duração de atuação e temperatura, expõem diferentes antígenos virais e devem ser previamente testados para cada sistema a ser estabelecido. Uma digestão mais intensa deve ser aplicada para a detecção do vírus do sarampo em tecido pulmonar do que para adenovírus ou vírus respiratório sincicial no mesmo tecido. Por outro lado, o vírus da febre amarela em tecido de fígado necessita de uma digestão mais fraca.

  16. A comparative study of an elisa test and an indirect immunofluorescence test for serological diagnosis of Babesia bovis infection

    International Nuclear Information System (INIS)

    Martins, J.R.; Cheong, F.H.; Correa, B.L.; Radley, D.E.; Cereser, V.H.

    1998-01-01

    Detection of antibodies to Babesia bovis in cattle is essential for the understanding of the epidemiology of babesiosis and this study was concerned with comparing the indirect fluorescent antibody with the ELISA. Both assays gave rise to 100% sensitivity whilst the ELISA was shown to be marginally more specific at 98%. The ease of use and low cost of the ELISA would make it the more obvious choice in conducting future serological surveys for this parasite. (author)

  17. Neuromyelitis optica immunoglobulin G in Chinese patients detected by immunofluorescence assay on a monkey brain substrate.

    Science.gov (United States)

    Long, Youming; Hu, Xueqiang; Peng, Fuhua; Lu, Zhengqi; Wang, Yuge; Yang, Yu; Qiu, Wei

    2012-01-01

    Serum neuromyelitis optica immunoglobulin G (NMO-IgG) is used as a biomarker to differentiate between neuromyelitis optica (NMO) and multiple sclerosis (MS). However, the original assay is expensive and complex and shows low sensitivity. Here, we investigated the potential of NMO-IgG detection using an indirect immunofluorescence (IIF) assay on monkey brains. NMO-IgG seroprevalence was determined in 168 samples by an IIF assay on a monkey brain substrate. The data were compared with those from a standard mouse brain IIF assay using McNemar and kappa tests. Thirty-one of 50 (62%) NMO patients, 7 of 18 (38.9%) longitudinally extensive transverse myelitis patients, 6 of 57 (10.5%) MS patients, and 5 of 10 (50%) optic neuritis patients were seropositive for NMO-IgG. None of the acute partial transverse myelitis patients (n = 3) or healthy controls (n = 20) was positive. Thus, the sensitivity of the test was 62% for the patients with clinically definite NMO. The specificity was 89.5%, considering the 57 MS patients as the control group. The modified IIF assay on monkey brains and the standard IIF assay based on mouse brains were not significantly different (McNemar test; p = 1.000). The two assays were concordant in 39 seropositive samples and 100 seronegative samples (kappa test; kappa = 0.592, p monkey brain assay was no better than the standard mouse brain IIF assay, we affirmed that NMO-IgG is a sensitive and specific biomarker to differentiate between NMO and MS. Copyright © 2011 S. Karger AG, Basel.

  18. Double-label immunofluorescence method for simultaneous detection of adenovirus and herpes simplex virus from the eye.

    Science.gov (United States)

    Walpita, P; Darougar, S

    1989-07-01

    The development and application of a double-label immunofluorescence method which has the potential to screen for single or dual infections from any site, in single shell vial cultures, is described. In this study, a total of 1,141 ocular specimens were inoculated in shell vials, centrifuged at 15,000 X g for 1 h, incubated at 37 degrees C for 48 h, and fixed in methanol at room temperature for 15 min. The virus inclusions were detected by staining with a double-label indirect immunofluorescence procedure using mixtures of appropriate first antibodies, followed by fluorescein- and rhodamine-conjugated second antibodies. Each specimen was also inoculated in parallel by the conventional virus isolation method. The sensitivity and specificity of the double-label shell vial procedure were comparable to those with the conventional method, and the former test took only 48 h to complete. The test offers a rapid and simple single-vial procedure which allows for individual or simultaneous detection of multiple pathogens. It results in savings in time and cost over the conventional virus isolation method and other shell vial procedures.

  19. Immunofluorescent detection in the ovary of host antibodies against a secretory ferritin injected into female Haemaphysalis longicornis ticks.

    Science.gov (United States)

    Galay, Remil Linggatong; Matsuo, Tomohide; Hernandez, Emmanuel Pacia; Talactac, Melbourne Rio; Kusakisako, Kodai; Umemiya-Shirafuji, Rika; Mochizuki, Masami; Fujisaki, Kozo; Tanaka, Tetsuya

    2018-04-01

    Due to the continuous threat of ticks and tick-borne diseases to human and animal health worldwide, and the drawbacks of chemical acaricide application, many researchers are exploring vaccination as an alternative tick control method. Earlier studies have shown that host antibodies can circulate in the ticks, but it has not been confirmed whether these antibodies can be passed on to the eggs. We previously reported that ticks infesting rabbits immunized with a recombinant secretory ferritin of Haemaphysalis longicornis (HlFER2) had reduced egg production and hatching. Here we attempted to detect the presence of antibodies against HlFER2 in the ovary and eggs of female ticks through immunofluorescent visualization. Purified anti-HlFER2 antibodies or rabbit IgG for control was directly injected to engorged female H. longicornis. Ovaries and eggs after oviposition were collected and prepared for an indirect immunofluorescent antibody test. Positive fluorescence was detected in ovaries one day post-injection of anti-HlFER2 antibodies. Through silencing of Hlfer2 gene, we also determined whether the injected antibodies can specifically bind to native HlFER2. Immunofluorescence was observed in the oocytes of dsLuciferase control ticks injected with anti-HlFER2 antibodies, but not in the oocytes of Hlfer2-silenced ticks also injected with anti-HlFER2 antibodies. Our current findings suggest that host antibodies can be passed on to the oocytes, which is significant in formulating a vaccine that can disrupt tick reproduction. Copyright © 2017 Elsevier B.V. All rights reserved.

  20. Indirect detection of dark matter

    International Nuclear Information System (INIS)

    Carr, J; Lamanna, G; Lavalle, J

    2006-01-01

    This article is an experimental review of the status and prospects of indirect searches for dark matter. Experiments observe secondary particles such as positrons, antiprotons, antideuterons, gamma-rays and neutrinos which could originate from annihilations of dark matter particles in various locations in the galaxy. Data exist from some experiments which have been interpreted as hints of evidence for dark matter. These data and their interpretations are reviewed together with the new experiments which are planned to resolve the puzzles and make new measurements which could give unambiguous results

  1. Do infants detect indirect reciprocity?

    Science.gov (United States)

    Meristo, Marek; Surian, Luca

    2013-10-01

    In social interactions involving indirect reciprocity, agent A acts prosocially towards B and this prompts C to act prosocially towards A. This happens because A's actions enhanced its reputation in the eyes of third parties. Indirect reciprocity may have been of central importance in the evolution of morality as one of the major mechanisms leading to the selection of helping and fair attitudes. Here we show that 10-month-old infants expect third parties to act positively towards fair donors who have distributed attractive resources equally between two recipients, rather than toward unfair donors who made unequal distributions. Infants' responses were dependent on the reciprocator's perceptual exposure to previous relevant events: they expected the reciprocator to reward the fair donor only when it had seen the distributive actions performed by the donors. We propose that infants were able to generate evaluations of agents that were based on the fairness of their distributive actions and to generate expectations about the social preferences of informed third parties. Copyright © 2013 Elsevier B.V. All rights reserved.

  2. Use of a Granulocyte Immunofluorescence Assay Designed for Humans for Detection of Antineutrophil Cytoplasmic Antibodies in Dogs with Chronic Enteropathies.

    Science.gov (United States)

    Florey, J; Viall, A; Streu, S; DiMuro, V; Riddle, A; Kirk, J; Perazzotti, L; Affeldt, K; Wagner, R; Vaden, S; Harris, T; Allenspach, K

    2017-07-01

    Perinuclear antineutrophil cytoplasmic antibodies (pANCA) previously have been shown to be serum markers in dogs with chronic enteropathies, with dogs that have food-responsive disease (FRD) having higher frequencies of seropositivity than dogs with steroid-responsive disease (SRD). The indirect immunofluorescence (IIF) assay used in previous publications is time-consuming to perform, with low interobserver agreement. We hypothesized that a commercially available granulocyte IIF assay designed for humans could be used to detect perinuclear antineutrophil cytoplasmic antibodies in dogs. Forty-four dogs with FRD, 20 dogs with SRD, 20 control dogs, and 38 soft-coated wheaten terrier (SCWT) or SCWT-cross dogs. A granulocyte assay designed for humans was used to detect pANCA, cANCA, and antinuclear antibodies (ANA), as well as antibodies against proteinase-3 protein (PR-3) and myeloperoxidase protein (MPO) in archived serum samples. Sensitivity of the granulocyte assay to predict FRD in dogs was 0.61 (95% confidence interval (CI), 0.45, 0.75), and specificity was 1.00 (95% CI, 0.91, 1.00). A significant association was identified between positive pANCA or cANCA result and diagnosis of FRD (P < 0.0001). Agreement between the two assays to detect ANCA in the same serum samples from SCWT with protein-losing enteropathy/protein-losing nephropathy (PLE/PLN) was substantial (kappa, 0.77; 95% CI, 0.53, 1.00). Eight ANCA-positive cases were positive for MPO or PR-3 antibodies. The granulocyte immunofluorescence assay used in our pilot study was easy and quick to perform. Agreement with the previously published method was good. Copyright © 2017 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.

  3. Detection of thrombocytic antibodies with the direct and indirect haemolysis inhibition test and the radioimmuno-Coombs test

    International Nuclear Information System (INIS)

    Mettenboerger, D.; Vith, E.

    1982-01-01

    Methods of application of the direct and indirect haemolysis inhibition test were studied in order to optimise the test parameters: The ultimate aim was to standardize the test method and compare its sensitivity in detecting various platelet antibodies with platelet indirect radioactive Coombs-test and the platelet immunofluorescence test. (orig.) [de

  4. An expert protocol for immunofluorescent detection of calcium channels in tsA-201 cells.

    Science.gov (United States)

    Koch, Peter; Herzig, Stefan; Matthes, Jan

    Pore-forming subunits of voltage gated calcium channels (VGCC) are large membrane proteins (260kDa) containing 24 transmembrane domains. Despite transfection with viral promoter driven vectors, biochemical analysis of VGCC is often hampered by rather low expression levels in heterologous systems rendering VGCC challenging targets. Especially in immunofluorescent detection, calcium channels are demanding proteins. We provide an expert step-by-step protocol with adapted conditions for handling procedures (tsA-201 cell culture, transient transfection, incubation time and temperature at 28°C or 37°C and immunostaining) to address the L-type calcium-channel pore Ca v 1.2 in an immunofluorescent approach. We performed immunocytochemical analysis of Ca v 1.2 expression at single-cell level in combination with detection of different markers for cellular organelles. We show confluency levels and shapes of tsA-201 cells at different time points during an experiment. Our experiments reveal sufficient levels of Ca v 1.2 protein and a correct Ca v 1.2 expression pattern in polygonal shaped cells already 12h after transfection. A sequence of elaborated protocol modifications allows subcellular localization analysis of Ca v 1.2 in an immunocytochemical approach. We provide a protocol that may be used to achieve insights into physiological and pathophysiological processes involving voltage gated calcium channels. Our protocol may be used for expression analysis of other challenging proteins and efficient overexpression may be exploited in related biochemical techniques requiring immunolabels. Copyright © 2016 Elsevier Inc. All rights reserved.

  5. Development of immunofluorescence colony staining (IFC) for detection of Xanthomonas campestris pv. vesicatoria and Clavibacter michiganensis subsp michiganensis in tomato seeds

    NARCIS (Netherlands)

    Nemeth, J.; Vuurde, van J.W.L.

    2006-01-01

    Immunofluorescence colony-staining (IFC) is based on sample pour plating in combination with immunofluorescence staining for recognition of the target colony. IFC was optimised for detecting Xanthomonas campestris pv. vesicatoria (Xcv) and Clavibacter michiganensis subsp. michiganensis (Cmm) in

  6. DETECTION OF ANTIBODIES TO CANDIDA ALBICANS GERM TUBE BY IMMUNOFLUORESCENCE IN IMMUNOSUPPRESSED MICE WITH EXPERIMENTAL SYSTEMIC CANDIDIASIS

    Directory of Open Access Journals (Sweden)

    F. Zaini

    2007-07-01

    Full Text Available "nThe increasing incidence of systemic candidiasis, which parallels the use of invasive and immunosuppressive medical procedures, necessitates development of rapid and cost effective tests for diagnosis of systemic candidiasis. Therefore in this study 85 mice were first immunosuppressed by cyclophosphamide and then infected by Candida albicans NCPF 3153. Other 85 mice were employed as control. The case and control mice were bled and then autopsied. Hearts and kidneys were checked by direct, histopathological and cultural examination for systemic candidiasis. The 85 sera from histological proven cases and 85 control mice were adsorbed with heat killed blastospores of same strain of C. albicans. Anti-Candida albicans germ tube antibodies were detected by indirect immunofluorescence assay for diagnosis of invasive candidiasis in case and control mice. In addition, sera from 35 mice with proven cryptococcosis were also tested. While 84 mice with proven systemic candidiasis (100% had anti-germ tube antibodies, these antibodies were absent in all controls and mice with cryptococcosis. The specificity was 100%, indicating a high degree of discrimination was possible between systemic candidiasis and cryptococcosis in the mice studied. It must be concluded that anti-germ tube responses did not appear to be significantly reduced in immunocompromised mice.

  7. Semi-automated, occupationally safe immunofluorescence microtip sensor for rapid detection of Mycobacterium cells in sputum.

    Directory of Open Access Journals (Sweden)

    Shinnosuke Inoue

    Full Text Available An occupationally safe (biosafe sputum liquefaction protocol was developed for use with a semi-automated antibody-based microtip immunofluorescence sensor. The protocol effectively liquefied sputum and inactivated microorganisms including Mycobacterium tuberculosis, while preserving the antibody-binding activity of Mycobacterium cell surface antigens. Sputum was treated with a synergistic chemical-thermal protocol that included moderate concentrations of NaOH and detergent at 60°C for 5 to 10 min. Samples spiked with M. tuberculosis complex cells showed approximately 10(6-fold inactivation of the pathogen after treatment. Antibody binding was retained post-treatment, as determined by analysis with a microtip immunosensor. The sensor correctly distinguished between Mycobacterium species and other cell types naturally present in biosafe-treated sputum, with a detection limit of 100 CFU/mL for M. tuberculosis, in a 30-minute sample-to-result process. The microtip device was also semi-automated and shown to be compatible with low-cost, LED-powered fluorescence microscopy. The device and biosafe sputum liquefaction method opens the door to rapid detection of tuberculosis in settings with limited laboratory infrastructure.

  8. Comparison of new immunofluorescence method for detection of soy protein in meat products with immunohistochemical, histochemical, and ELISA methods

    Directory of Open Access Journals (Sweden)

    Michaela Petrášová

    2014-01-01

    Full Text Available Soy proteins are commonly used in the food industry thanks to their technological properties. However, soy is, along with cow’s milk, eggs, wheat, peanuts, tree nuts, fish, crustaceans, and molluscs, responsible for around 90% of food allergies, and is also one of the foodstuffs that can cause anaphylaxis. The aim of this work was to compare the immunofluorescence method for the detection of soy protein in meat products purchased from the retail market with other microscopic methods (immunohistochemical and histochemical, with the ELISA reference method and with the confirmatory results. Within the research, 127 meat products purchased in the retail network were examined using the immunofluorescence method used for the detection of soy protein. The method was compared to Enzyme-Linked ImmunoSorbent Assay (ELISA, immunohistochemical, and histochemical methods. According to McNemar’s test, non-compliance between the immunofluorescence method and immunohistochemical method was low. In addition, a significant difference between the fluorescence method and ELISA (P P < 0.01 was found. The immunofluorescence method was also compared with confirmatory results. According to McNemar’s test, non-compliance between the immunofluorescence method and confirmatory results was low. The results showed the possibilities of this new method to detect the content of soy protein in meat products.

  9. Estimation of antibodies to human cytomegalovirus by immunofluorescence and radioimmunoassay

    International Nuclear Information System (INIS)

    Jankowski, M.; Gut, W.; Nawrocka, E.

    1980-01-01

    The 125 I labelled IgG fraction against rabbit IgG of goat origin was employed for the detection of CMV IgG and IgM antibodies in the double indirect radioimmunoassay. The results were compared with those obtained in complement fixation, indirect immunofluorescence and anti-complement immunofluorescence tests. The application of labelled anti-fc antisera, instead of antisera against whole IgG in the tests for detection of specific CMV IgG antibody resulted in increased sensitivity of radioimmunoassay and reduction of non-specific cytoplasmatic reactions in preparations stained by indirect immunofluorescence. The absorption of sera with protein A rich staphylococci and aggregates to immunoglobulin eliminated unwanted secondary IgM staining caused by rheumatoid factors both in indirect immunofluorescence and radioimmunoassay tests for CMV antibodies. (author)

  10. Indirect detection methods: Looking for what is not there

    International Nuclear Information System (INIS)

    Yeung, E.S.

    1989-01-01

    Indirect detection allows one to monitor species that normally do not give a response. It has grown from an intellectual curiosity to become an extremely useful tool. For volume displacement, polarimetry gives the best results. For charge displacement, ion chromatography and capillary zone electrophoresis are ideal situations for applying indirect fluorescence detection. LOD surpasses all but the most specialized detection schemes. Future developments in systems specially designed for indirect detection should make these schemes even more powerful

  11. Spherical cows in dark matter indirect detection

    Energy Technology Data Exchange (ETDEWEB)

    Bernal, Nicolás [Centro de Investigaciones, Universidad Antonio Nariño, Cra 3 Este # 47A-15, Bogotá (Colombia); Necib, Lina; Slatyer, Tracy R., E-mail: nicolas.bernal@uan.edu.co, E-mail: lnecib@mit.edu, E-mail: tslatyer@mit.edu [Center for Theoretical Physics, Massachusetts Institute of Technology, Cambridge, MA 02139 (United States)

    2016-12-01

    Dark matter (DM) halos have long been known to be triaxial, but in studies of possible annihilation and decay signals they are often treated as approximately spherical. In this work, we examine the asymmetry of potential indirect detection signals of DM annihilation and decay, exploiting the large statistics of the hydrodynamic simulation Illustris. We carefully investigate the effects of the baryons on the sphericity of annihilation and decay signals for both the case where the observer is at 8.5 kpc from the center of the halo (exemplified in the case of Milky Way-like halos), and for an observer situated well outside the halo. In the case of Galactic signals, we find that both annihilation and decay signals are expected to be quite symmetric, with axis ratios very different from 1 occurring rarely. In the case of extragalactic signals, while decay signals are still preferentially spherical, the axis ratio for annihilation signals has a much flatter distribution, with elongated profiles appearing frequently. Many of these elongated profiles are due to large subhalos and/or recent mergers. Comparing to gamma-ray emission from the Milky Way and X-ray maps of clusters, we find that the gamma-ray background appears less spherical/more elongated than the expected DM signal from the large majority of halos, and the Galactic gamma ray excess appears very spherical, while the X-ray data would be difficult to distinguish from a DM signal by elongation/sphericity measurements alone.

  12. Evaluation of immunofluorescence microscopy and enzyme-linked immunosorbent assay in detection of Cryptosporidium and Giardia infections in asymptomatic dogs

    DEFF Research Database (Denmark)

    Rimhanen-Finne, R.; Enemark, Heidi L.; Kolehmainen, J.

    2007-01-01

    The performance of immunofluorescence microscopy (IF) and enzyme-linked immunosorbent assay (ELISA) in canine feces was evaluated. IF and Cryptosporidium ELISA detected 10(5) oocysts/g, while the detection limit for Giardia ELISA was 10(4) cysts/g. The Cryptosporidium ELISA showed 94% specificity...... zoonotic character of Cryptosporidium and Giardia infections in 150 asymptomatic Finnish dogs from the Helsinki area were studied. The overall proportion of dogs positive for Cryptosporidium was 5% (7/150) and that for Giardia 5% (8/150). In dogs...

  13. Direct and indirect detection of supersymmetric dark matter; Detection directe et indirecte de matiere sombre supersymetrique

    Energy Technology Data Exchange (ETDEWEB)

    Mayet, F

    2001-09-01

    A substantial body of astrophysical evidence supports the existence of non-baryonic dark matter in the universe. One of the leading dark matter candidates is the neutralino predicted by the supersymmetric extensions of the standard model of particle physics. Different detectors have been designed for the detection, either indirect or direct, of the neutralino. Related to indirect detection, the present work has been performed in the context of the AMS experiment. A precursor version of the spectrometer was flown on the space shuttle Discovery in June 1998. The detector included an Aerogel Threshold Cherenkov counter (ATC) to identify antiprotons, whose spectrum may be used to infer a neutralino signal. The analysis of the ATC data is presented including an evaluation of the flight performance and a description of the optimization of the antiproton selection. An antiproton analysis is also reported. A phenomenological study allows us to investigate the discovery potential of this indirect method. This thesis also includes the development of a new detector (MACHe3) designed for direct neutralino search using a superfluid {sup 3}He bolometer operated at ultra low temperatures. The data analysis of the prototype cell is presented. A Monte Carlo simulation has been developed, in order to optimize the detector design for direct neutralino search. These results are compared with theoretical predictions of supersymmetric models, thus highlighting the discovery potential of this detector and its complementarity with existing devices. (author)

  14. A Unique Immunofluorescence Protocol to Detect Protein Expression in Vascular Tissues: Tacking a Long Standing Pathological Hitch

    Directory of Open Access Journals (Sweden)

    Puneet GANDHI

    2018-01-01

    Full Text Available Objective: Autofluorescence induced interference is one of the major drawbacks in immunofluorescence analysis of formalin-fixed paraffin-embedded tissues, as it decreases the signal-to-noise ratio of specific labeling. Apart from aldehyde-fixation induced artifacts; collagen and elastin, red blood cells and endogenous fluorescent pigment lipofuscin are prime sources of autofluorescence in vascular and aging tissues. We describe herein, an optimized indirect-immunofluorescence method for archival formalin-fixed paraffin-embedded tissues tissues and cryo sections, using a combination of 3-reagents in a specific order, to achieve optimal fluorescence signals and imaging. Material and Method: Human telomerase reverse transcriptase, a protein implicated as a proliferation marker, was chosen relevant to its expression in solid tumors along with 3 other intracellular proteins exhibiting nuclear and/or cytoplasmic expression. Staining was performed on 10 glioma tissue sections along with 5 of their cryo sections, 5 sections each of hepatocellular, lung, papillary-thyroid and renal cell carcinoma, with 10 non-malignant brain tissue samples serving as control. Specimens were imaged using epifluorescence microscopy, followed by software-based quantification of fluorescence signals for statistical analysis and validation. Results: We observed that the combined application of sodium-borohydride followed by crystal violet before antigen retrieval and a Sudan black B treatment after secondary antibody application proved to be most efficacious for masking autofluorescence/non-specific background in vascular tissues. Conclusion: This unique trio-methodology provides quantifiable observations with maximized fluorescence signal intensity of the target protein for longer retention time of the signal even after prolonged storage. The results can be extrapolated to other human tissues for different protein targets.

  15. Practical application of immunofluorescence for the detection of bacterial contaminants during vinification

    Directory of Open Access Journals (Sweden)

    Marielle Bouix

    1997-03-01

    Full Text Available This study presents a rapid and specific microscopie technique for detecting and identifying populations of lactic acid bacteria in musts, wines, and inoculum starter cultures. Through the use of fluorescent antibodies, this procedure can be performed in less than two hours, and it is effective with Leuconostoc, Pediococcus and Lactobacillus concentrations as small as 102 cells/ml. Implementation of this technique will assist winemakers in controlling malolactic fermentations and in preventing lactic acid bacterial spoilage.

  16. Naturally acquired Lawsonia intracellularis infection in pigs studied from weaning to slaughter by indirect immunofluorescence antibody test and polymerase chain reaction on faeces

    DEFF Research Database (Denmark)

    Jensen, Tim Kåre; Vigre, Håkan; Sørensen, Vibeke

    2005-01-01

    by immunofluorescence antibody test compared to 24% by immunohistochemistry on ileal samples. Thus, applied at the time of slaughter the antibody test appeared to be a highly sensitive ante-mortem diagnostic tool for identifying L. intracelluaris exposed pigs with or without current proliferative enteropathy. (c) 2004......The course of naturally acquired Lawsonia intracellularis infection was studied in 41 pigs by testing blood and faeces samples collected four to seven times from before weaning to slaughter 5 months old. At slaughter, a sample of ileum was taken for histopathology. In the first sampling when...... later on shed and/or were seropositive for L. intracellularis. The lowest prevalence of L. intracellularis was observed in 6-13 weeks old pigs and it seemed as though L. intracellularis in early infected pigs only activates a minor antibody response. At slaughter 66% of the pigs were found positive...

  17. Reações de imunofluorescência indireta: algumas modificações de sua técnica Indirect immunofluorescence tests: some modifications of their technique

    Directory of Open Access Journals (Sweden)

    Cláudio Santos Ferreira

    1973-09-01

    Full Text Available Propõem-se modificações de procedimentos rotineiros de reações de imunofluorescência indireta. A eluição das amostras de sangue dessecado contidas em papel-filtro é feita em placas ("godets" com tampas. Alças de níquel-cromo substituem pipetas capilares para a transferência de amostras de eluatos para as lâminas de microscopia. Propõe-se uma técnica rápida e eficiente de lavagem das preparações.Some simplifications intended for routine operations included immunofluorescence tests are proposed. The elution of dried blood samples is performed on test plates (water-color mixing plates, with a lid instead of in test tubes. Nickel-chromium wire loops are substituted for capillary pipettes in transferring eluate samples to microscope slides. A rapid and efficient washing technique is proposed.

  18. Evaluation of a Direct Immunofluorescent Assay and/or Conjunctival Cytology for Detection of Canine Distemper Virus Antigen.

    Science.gov (United States)

    Athanasiou, Labrini V; Kantere, Maria C; Kyriakis, Constantinos S; Pardali, Dimitra; Adamama Moraitou, Katerina; Polizopoulou, Zoe S

    2018-04-01

    Canine distemper is a common and potentially lethal multisystemic disease caused by the Canine distemper virus (CDV). We evaluated the diagnostic performance of direct immunofluorescent assay (FA) and cytology to detect CDV antigen in conjunctival cells compared with an established polymerase chain reaction (PCR) detection assay used as a gold standard for CDV diagnosis. Samples were collected from 57 young dogs presenting with central nervous system signs compatible with distemper disease. Exfoliative epithelial cells were collected from the right and left conjunctiva of each animal using nylon-bristled cytobrushes for cytology and cotton swabs for FA and PCR. For the direct FA, samples were stained with anti-CDV polyclonal antiserum conjugated to fluorescein isothiocyanate and imaged using a fluorescent microscope. Out of 57 dogs tested, 19 were PCR positive (15 positive in direct FA and 4 positive in cytology, including one that was negative by PCR), whereas 37 dogs were negative in all methods. A good agreement was observed between the FA and PCR, with a κ-value of 0.833 (95% CI: 0.678-0.989). Meanwhile, there was poor agreement between cytology and PCR (κ-value of 0.164; 95% CI: -0.045 to 0.373) and a fair agreement between FA and cytology (κ-value of 0.231; 95% CI: -0.026 to 0.487). Our results indicated a poor performance of cytology for the detection of CDV antigen. In contrast, FA is a 100% specific and an adequately sensitive assay (sensitivity: 78.95%, negative likelihood ratio: 0.21, 95% CI: 0.09-0.50) for antemortem diagnosis of canine distemper.

  19. Diagnostic utility of a direct immunofluorescence test to detect feline coronavirus antigen in macrophages in effusive feline infectious peritonitis.

    Science.gov (United States)

    Litster, A L; Pogranichniy, R; Lin, T-L

    2013-11-01

    The antemortem diagnosis of feline infectious peritonitis (FIP) remains challenging in clinical practice, since current testing methods have suboptimal diagnostic accuracy. Immunohistochemical testing of biopsy specimens and postmortem examination are the standard diagnostic methods, although direct immunofluorescence (DIF) testing to detect feline coronavirus in macrophages in effusion specimens has been reported to have 100% specificity and has been recommended as an antemortem confirmatory test. The aim of this study was to compare the results of DIF testing in antemortem feline effusions with postmortem results using field samples. Effusion specimens were collected antemortem from 17 cats and tested by DIF, followed by postmortem examination. Histopathological examination of specimens collected at postmortem confirmed FIP in 10/17 cases and ruled out FIP out in 7/17 cases. Antemortem DIF testing was positive in all 10 cases confirmed as FIP at postmortem examination. In the seven cats where FIP was ruled out at postmortem examination, DIF was negative in five cases and positive in the remaining two cases. The calculated sensitivity of DIF testing was 100% and the specificity was 71.4%. Duplicate effusion specimens from eight cats that were initially DIF positive were stored refrigerated (4 °C) or at room temperature (22-25 °C) and subjected to serial DIF testing to determine the duration of positive results. DIF-positive specimens stored at both temperatures retained their positive status for at least 2 days. Copyright © 2013 Elsevier Ltd. All rights reserved.

  20. The detection of Giardia muris and Giardia lamblia cysts by immunofluorescence in animal tissues and fecal samples subjected to cycles of freezing and thawing.

    Science.gov (United States)

    Erlandsen, S L; Sherlock, L A; Bemrick, W J

    1990-04-01

    The effects of freezing and thawing on the detection of selected Giardia spp. cysts were investigated using immunofluorescence, bright field microscopy, and low voltage scanning electron microscopy (SEM). Giardia muris cysts were obtained from either animal carcasses, fecal pellets, or isolated cyst preparations, whereas Giardia lamblia cysts were isolated from fecal samples. These samples were stained using an immunofluorescence technique after 1-3 freezing (-16 C) and thawing (20 C) cycles. Cysts were detected successfully by immunofluorescence in all samples. However, in those samples subjected to freeze-thawing, the cyst walls often became distorted and then were not detectable by bright field microscopy. Low voltage SEM demonstrated that the filaments in the distorted cyst wall underwent rearrangements of interfilament spacing. Quantitation of cyst recovery after freezing and thawing demonstrated that a substantial loss occurred after 1 cycle of alternating temperature when low concentrations of cysts were used, but not with high concentrations of cysts. Cyst recovery, after 3 freezing and thawing cycles, was dramatically lowered irrespective of the initial cyst concentration. These results demonstrated that immunofluorescence was an effective technique for the detection of Giardia spp. cysts in frozen samples and would suggest that freezing and thawing of fecal samples could prevent the detection of cysts when only bright field microscopy was employed.

  1. Direct versus indirect detection of supersymmetric dark matter

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    2003-07-01

    This document gathers the slides that were presented during the workshop 'direct versus indirect detection of supersymmetric dark matter'(about 30 contributions). This workshop intended to bring together people from the particle theory community, astrophysicists and cosmologists, as well as experimentalists involved in the detection of dark matter. The aim is to generate a discussion about current and future strategies for detection of SUSY dark matter (with focus, but not exclusively, on neutralinos). Complementarities between accelerator, direct and indirect searches as well as a comparison between the uncertainties in direct and indirect searches of dark matter, are supposed to be discussed. Among the issues which will be addressed are: -) the crucial questions related to the structure of galaxies (local dark matter density, clumping, anomalous velocity distributions, etc.) ; -) the possibilities offered by the present and future experimental facilities for direct and indirect (photon, neutrino) searches; -) the potential for the discovery of SUSY at LHC and beyond; and -) the parameterization of the SUSY breaking models beyond the minimal versions.

  2. Direct versus indirect detection of supersymmetric dark matter

    International Nuclear Information System (INIS)

    2003-01-01

    This document gathers the slides that were presented during the workshop 'direct versus indirect detection of supersymmetric dark matter'(about 30 contributions). This workshop intended to bring together people from the particle theory community, astrophysicists and cosmologists, as well as experimentalists involved in the detection of dark matter. The aim is to generate a discussion about current and future strategies for detection of SUSY dark matter (with focus, but not exclusively, on neutralinos). Complementarities between accelerator, direct and indirect searches as well as a comparison between the uncertainties in direct and indirect searches of dark matter, are supposed to be discussed. Among the issues which will be addressed are: -) the crucial questions related to the structure of galaxies (local dark matter density, clumping, anomalous velocity distributions, etc.) ; -) the possibilities offered by the present and future experimental facilities for direct and indirect (photon, neutrino) searches; -) the potential for the discovery of SUSY at LHC and beyond; and -) the parameterization of the SUSY breaking models beyond the minimal versions

  3. [Immunofluorescence assay with Crithidia luciliae for the detection of anti-DNA antibodies. Atypical images and their relationship with Chagas' disease and leishmaniasis].

    Science.gov (United States)

    Griemberg, Gloria; Ferrarotti, Nidia F; Svibel, Graciela; Ravelli, Maria R; Taranto, Nestor J; Malchiodi, Emilio L; Pizzimenti, Maria C

    2006-01-01

    Anti-native DNA antibodies can be detected by indirect immunofluorescence assay with Crithidia luciliae, displaying an annular image due to a kinetoplast containing double stranded DNA. Other structures such as membrane, flagellum and basal corpuscle can be stained as well, showing what is called atypical fluorescent images. As C. luciliae belongs to the Trypanosomatidae family, which include the human pathogens Trypanosoma cruzi and Leishmania spp., it was considered that these atypical images could be caused by cross-reactions. Serological studies for Chagas' disease were performed in 105 serum samples displaying atypical images. Sixty four percent of the samples from non endemic and 78.3% from endemic areas for Chagas' disease showed fluorescence in both, membrane and flagellum (joint image). Fifty samples from normal blood donors and 57 samples from patients with conective tissue diseases were tested with C. luciliae. None of them presented the joint image except for two patients with lupus who were also chagasic. In addition, 54 samples from chagasic patients were studied and all of them presented the joint image. We also studied 46 samples from patients with leishmaniasis from whom 28 were coinfected with T. cruzi. The joint image was observed in 88.0% of the samples with leishmaniasis and in 89.3% of the co-infected samples. The results suggest that C. luciliae could be used as an economical, and of low risk, alternative substrate for the serological diagnosis of Chagas' disease, even though it does not discriminate for Leishmania spp. infection. This study also suggests that whenever atypical images are observed in C. luciliae during the search for anti-DNA antibodies, it would be convenient to submit the patient to clinical and serological tests for the diagnosis of leishmaniosis and Chagas' disease.

  4. [Application of Immunohistochemistry and Immunofluorescence Staining in Detection of Phospholipase A2 Receptor on Paraffin Section of Renal Biopsy Tissue].

    Science.gov (United States)

    Dong, Hong-rui; Wang, Yan-yan; Wang, Guo-qin; Sun, Li-jun; Cheng, Hong; Chen, Yi-pu

    2015-10-01

    To evaluate the application of immunohistochemistry and fluorescence staining method in the detection of phospholipase A2 receptor (PLA2R) on paraffin section of renal biopsy tissue,and to find an accurate and fast method for the detection of PLA2R in renal tissue. The PLA2R of 193 cases were detected by immunohistochemical staining,and the antigen was repaired by the method of high pressure cooker (HPC) hot repair plus trypsin repair. The 193 samples including 139 cases of idiopathic membranous nephropathy (IMN), 15 cases of membranous lupus nephritis, 8 cases of hepatitis B virus associated membranous nephropathy, 18 cases of IgA nephropathy, and 13 cases of minimal change diseases. To compare the dyeing effects, 22 paraffin sections of renal biopsy tissue of IMN cases with positive PLA2R were stained by using 4 different. of antigen repairing,which included HPC hot repair, HPC hot repair plus trypsin repair, water bath heat repair, and water bath heat repair plus trypsin repair. To compare the dyeing effects, 15 paraffin sections of renal biopsy tissue of IMN cases with positive PLA2R were stained by using 3 different. of antigen repairing,which included water bath heat repair plus trypsin repair, protease K digestion repair, and pepsin digestion repair. In 193 cases, the positive rate of PLA2R in IMN cases was 90.6% (126/139), and the other 54 patients without IMN were negative. Twenty-two IMN patients were positive for PLA2R by using the HPC heat repair plus trypsin repaire or the water bath heat repair plus trypsin repair;while only a few cases of 22 IMN cases were positive by using the HPC hot repair alone or water bath heat repair alone. Fifteen IMN patients were positive for PLA2R by using water bath heat repair plus trypsin repair,protease K digestion repair,and pepsin digestion repair, but the distribution of positive deposits and the background were different. PLA2R immunohistochemical staining can effectively identify IMN and secondary MN. For

  5. Padronização de ensaio imunoenzimático para pesquisa de anticorpos das classes IgM e IgG anti-Toxoplasma gondii e comparação com a técnica de imunofluorescência indireta Standardization of enzyme-linked immunosorbent assay ELISA to detect anti-Toxoplasma gondii IgM and IgG antibodies, and comparison with the indirect immunofluorescence technique

    Directory of Open Access Journals (Sweden)

    Cláudia Maria Antunes Uchôa

    1999-12-01

    Full Text Available A sorologia tem sido o método de escolha para o diagnóstico da toxoplasmose. Devido a isto, padronizamos um ensaio imunoenzimático (ELISA e comparamos seus resultados com a técnica de imunofluorescência indireta (IFI. A técnica padronizada apresentou na pesquisa de IgG sensibilidade (S de 96,7% e especificidade (E de 75%, com valor de predição de positividade (VPP de 83,3% e de negatividade (VPN de 94,7%, com uma concordância ajustada (K de 73,5%. A IFI apresentou S de 83,8%, E de 79,1% com VPP de 83,8 % e VPN de 79,1% com K de 63%. A concordância bruta entre os dois testes (ELISA/IFI foi de 88,3% para pesquisa de IgG e de 81,5% para pesquisa de IgM, sendo o K de 70,8% para IgG e de 1,3% para IgM, sendo o índice de correlação (r de 0,556 para IgG e de -0,023 para IgM. Podemos concluir que a ELISA-IgG padronizada é indicada nos processos de triagem sorológica, sendo a ELISA-IgM desaconselhada uma vez que apresentou baixos índices de concordância ajustada com a técnica de referência, sugerindo pouca confiabilidade dos resultados.Serology has been the most popular method to diagnose toxoplasmosis. Accordingly, this study standardizes an enzyme-linked immunosorbent assay (ELISA and compares its results with the IFI technique. In the IgG detection test, the standardized technique presented a sensibility (S of 96.77%, a specificity (SP of 75%, with a positive predictive value (PPV of 83.33%, a negative predictive value (NPV of 94.74%, and an adjusted concordance (K of 73.50%. The IFI exhibited 83.87% for S, 79.16% for SP, 83.81% for PPV, 79.16% for NPV, and 63% for K. The rough concordance between these two tests (ELISA/IFI was 88.35% for the IgG detection test and 81.55% for the IgM detection test. K was 70.82% and 1.31% for IgG and IgM, respectively, the correlation index (r being 0.556 for IgG and -0.023 for IgM. We can conclude that standardized ELISA-IgG is indicated in serologic selection processes, whereas the ELISA-IgM is

  6. Desarrollo de improntas para el diagnóstico del virus Epstein-Barr por inmunofluorescencia indirecta Development of slides for Epstein-Barr virus diagnosis by indirect immunofluorescence

    Directory of Open Access Journals (Sweden)

    Germán R. Pérez

    2005-08-01

    mononucleosis. Detection of anti-viral capside antigen (VCA antibodies indicates previous or present EBV infection. Moreover, high titles of anti-VCA antibodies are observed in EBV-associated neoplasic disorders, such as lymphomas in AIDS patients. The objective of this study was the development and optimization of P3HR1 cell slides for the EBV serologic detection by indirect immunofluorescence (IIF assay. P3HR1 exponential growth culture cells were stimulated with phorbol-12-mirystoil-13-acetate, collected at different time points and used for slide preparation. IIF assay was performed in each slide using an anti-EBV positive serum as primary antibody. An 11% increase in VCA expression was observed at 40 hours post-stimulation. Data was confirmed by western blot and immunodetection. Intra- and inter-lot precisions of the developed slides were evaluated for IgG and IgM antibodies using EBV-positive sera and positive samples for other members of the Herpesviridae family. Neither false-positive or false negative results were obtained for EBV detection nor cross-reaction was observed with other members of the Herpesviridae family with the developed slides. In conclusion, the slides here presented can be a useful instrument for acute EBV infection diagnosis and for the serologic detection of IgG anti-VCA antibodies in EBV-associated neoplastic disorders.

  7. Review of indirect detection of dark matter with neutrinos

    Science.gov (United States)

    Danninger, Matthias

    2017-09-01

    Dark Matter could be detected indirectly through the observation of neutrinos produced in dark matter self-annihilations or decays. Searches for such neutrino signals have resulted in stringent constraints on the dark matter self-annihilation cross section and the scattering cross section with matter. In recent years these searches have made significant progress in sensitivity through new search methodologies, new detection channels, and through the availability of rich datasets from neutrino telescopes and detectors, like IceCube, ANTARES, Super-Kamiokande, etc. We review recent experimental results and put them in context with respect to other direct and indirect dark matter searches. We also discuss prospects for discoveries at current and next generation neutrino detectors.

  8. Indirect Detection Analysis: Wino Dark Matter Case Study

    Energy Technology Data Exchange (ETDEWEB)

    Hryczuk, Andrzej [Munich, Tech. U.; Cholis, Ilias [Fermilab; Iengo, Roberto [SISSA, Trieste; Tavakoli, Maryam [IPM, Tehran; Ullio, Piero [INFN, Trieste

    2014-07-15

    We perform a multichannel analysis of the indirect signals for the Wino Dark Matter, including one-loop electroweak and Sommerfeld enhancement corrections. We derive limits from cosmic ray antiprotons and positrons, from continuum galactic and extragalactic diffuse γ-ray spectra, from the absence of γ-ray line features at the galactic center above 500 GeV in energy, from γ-rays toward nearby dwarf spheroidal galaxies and galaxy clusters, and from CMB power-spectra. Additionally, we show the future prospects for neutrino observations toward the inner Galaxy and from antideuteron searches. For each of these indirect detection probes we include and discuss the relevance of the most important astrophysical uncertainties that can impact the strength of the derived limits. We find that the Wino as a dark matter candidate is excluded in the mass range bellow simeq 800 GeV from antiprotons and between 1.8 and 3.5 TeV from the absence of a γ-ray line feature toward the galactic center. Limits from other indirect detection probes confirm the main bulk of the excluded mass ranges.

  9. Indirect detection of radiation sources through direct detection of radiolysis products

    Science.gov (United States)

    Farmer, Joseph C [Tracy, CA; Fischer, Larry E [Los Gatos, CA; Felter, Thomas E [Livermore, CA

    2010-04-20

    A system for indirectly detecting a radiation source by directly detecting radiolytic products. The radiation source emits radiation and the radiation produces the radiolytic products. A fluid is positioned to receive the radiation from the radiation source. When the fluid is irradiated, radiolytic products are produced. By directly detecting the radiolytic products, the radiation source is detected.

  10. Direct and Indirect Dark Matter Detection in Gauge Theories

    Energy Technology Data Exchange (ETDEWEB)

    Queiroz, Farinaldo [Federal Univ. of Paraba (Brazil)

    2013-01-01

    The Dark matter (DM) problem constitutes a key question at the interface among Particle Physics, Astrophysics and Cosmology. The observational data which have been accumulated in the last years point to an existence of non baryonic amount of DM. Since the Standard Model (SM) does not provide any candidate for such non-baryonic DM, the evidence of DM is a major indication for new physics beyond the SM. We will study in this work one of the most popular DM candidates, the so called WIMPs (Weakly Interacting Massive Particles) from a direct and indirect detection perspective. In order to approach the direct and indirect dection of DM in the context of Particle Physics in a more pedagogic way, we will begin our discussion talking about a minimal extension of the SM. Later we will work on the subject in a 3-3-1 model. Next, we will study the role of WIMPs in the Big Bang Nucleosynthesis. Lastly, we will look for indirect DM signals in the center of our galaxy using the NASA Satellite, called Fermi-LAT. Through a comprehensive analysis of the data events observed by Fermi-LAT and some background models, we will constrain the dark matter annihilation cross section for several annihilation channels and dark matter halo profiles.

  11. Indirect detection of dark matter with γ rays.

    Science.gov (United States)

    Funk, Stefan

    2015-10-06

    The details of what constitutes the majority of the mass that makes up dark matter in the Universe remains one of the prime puzzles of cosmology and particle physics today-80 y after the first observational indications. Today, it is widely accepted that dark matter exists and that it is very likely composed of elementary particles, which are weakly interacting and massive [weakly interacting massive particles (WIMPs)]. As important as dark matter is in our understanding of cosmology, the detection of these particles has thus far been elusive. Their primary properties such as mass and interaction cross sections are still unknown. Indirect detection searches for the products of WIMP annihilation or decay. This is generally done through observations of γ-ray photons or cosmic rays. Instruments such as the Fermi large-area telescope, high-energy stereoscopic system, major atmospheric gamma-ray imaging Cherenkov, and very energetic radiation imaging telescope array, combined with the future Cherenkov telescope array, will provide important complementarity to other search techniques. Given the expected sensitivities of all search techniques, we are at a stage where the WIMP scenario is facing stringent tests, and it can be expected that WIMPs will be either be detected or the scenario will be so severely constrained that it will have to be rethought. In this sense, we are on the threshold of discovery. In this article, I will give a general overview of the current status and future expectations for indirect searches of dark matter (WIMP) particles.

  12. Unstable gravitino dark matter prospects for indirect and direct detection

    International Nuclear Information System (INIS)

    Grefe, Michael

    2011-11-01

    We confront the signals expected from unstable gravitino dark matter with observations of indirect dark matter detection experiments in all possible cosmic-ray channels. For this purpose we calculate in detail the gravitino decay widths in theories with bilinear violation of R parity, particularly focusing on decay channels with three particles in the final state. Based on these calculations we predict the fluxes of gamma rays, charged cosmic rays and neutrinos expected from decays of gravitino dark matter. Although the predicted spectra could in principal explain the anomalies observed in the cosmic ray positron and electron fluxes as measured by PAMELA and Fermi LAT, we find that this possibility is ruled out by strong constraints from gamma-ray and antiproton observations. Therefore, we employ current data of indirect detection experiments to place strong constraints on the gravitino lifetime and the strength of R-parity violation. In addition, we discuss the prospects of forthcoming searches for a gravitino signal in the spectrum of cosmic-ray antideuterons, finding that they are in particular sensitive to rather low gravitino masses. Finally, we discuss in detail the prospects for detecting a neutrino signal from gravitino dark matter decays, finding that the sensitivity of neutrino telescopes like IceCube is competitive to observations in other cosmic ray channels, especially for rather heavy gravitinos. Moreover, we discuss the prospects for a direct detection of gravitino dark matter via R-parity violating inelastic scatterings off nucleons. We find that, although the scattering cross section is considerably enhanced compared to the case of elastic gravitino scattering, the expected signal is many orders of magnitude too small in order to hope for a detection in underground detectors. (orig.)

  13. Unstable gravitino dark matter prospects for indirect and direct detection

    Energy Technology Data Exchange (ETDEWEB)

    Grefe, Michael

    2011-11-15

    We confront the signals expected from unstable gravitino dark matter with observations of indirect dark matter detection experiments in all possible cosmic-ray channels. For this purpose we calculate in detail the gravitino decay widths in theories with bilinear violation of R parity, particularly focusing on decay channels with three particles in the final state. Based on these calculations we predict the fluxes of gamma rays, charged cosmic rays and neutrinos expected from decays of gravitino dark matter. Although the predicted spectra could in principal explain the anomalies observed in the cosmic ray positron and electron fluxes as measured by PAMELA and Fermi LAT, we find that this possibility is ruled out by strong constraints from gamma-ray and antiproton observations. Therefore, we employ current data of indirect detection experiments to place strong constraints on the gravitino lifetime and the strength of R-parity violation. In addition, we discuss the prospects of forthcoming searches for a gravitino signal in the spectrum of cosmic-ray antideuterons, finding that they are in particular sensitive to rather low gravitino masses. Finally, we discuss in detail the prospects for detecting a neutrino signal from gravitino dark matter decays, finding that the sensitivity of neutrino telescopes like IceCube is competitive to observations in other cosmic ray channels, especially for rather heavy gravitinos. Moreover, we discuss the prospects for a direct detection of gravitino dark matter via R-parity violating inelastic scatterings off nucleons. We find that, although the scattering cross section is considerably enhanced compared to the case of elastic gravitino scattering, the expected signal is many orders of magnitude too small in order to hope for a detection in underground detectors. (orig.)

  14. Propagation of Galactic Cosmic Rays and Dark Matter indirect Detection

    International Nuclear Information System (INIS)

    Delahaye, Timur

    2010-07-01

    This thesis is dedicated to the study of propagation of cosmic electrons and positrons in the Milky Way and to the indirect detection of dark matter. The existence of dark matter is a hypothesis considered as reasonable from the point of view of cosmology, astrophysics and even particle physics. Nevertheless its detection still eludes us and it is not possible to verify this hypothesis by other means than gravitational one. A possible way to detect dark matter is to look for its annihilation or decay products among Galactic cosmic rays. During the last three years, data concerning cosmic ray electrons and positrons have been accumulated and have reached a remarkable precision. Such a precision requires from us to refine the theoretical models and to quantify the errors. This thesis addresses the study of all the sources of uncertainties affecting predictions of cosmic electrons and positron fluxes, primary and secondary, classical or from exotic origin. The greatest care has been dedicated to the sources and the propagation in the Galactic halo. Moreover a study of gamma and radio emissions associated to these cosmic rays is presented, again with the will of sizing uncertainties. Finally a status of the research for detection of annihilation or decay of Galactic dark matter is presented. (author)

  15. Anatomopatologia e imunofluorescência direta e indireta das lesões de pênfigo foliáceo endêmico resistentes à corticoterapia Pathology, direct and indirect immunofluorescent test of lesions of pemphigus foliaceous resistent to corticotherapy

    Directory of Open Access Journals (Sweden)

    Vitor Manoel Silva dos Reis

    1991-04-01

    Full Text Available São estudados 16 doentes de pênfigo foliáceo endêmico (PFE, sob tratamento com corticosteróides, que ainda apresentavam lesões eritemato-pápulo-verrucosas, geralmente hiperpigmentadas, que foram caracterizadas como lesões resistentes à corticoterapia (LRC. O estudo destas lesões foi feito através de anatomopatologia e de imunofluorescência direta (IFD. Anatomopatologicamente essas lesões mostraram tendência à hiperplasia epitelial e clivagem em níveis variáveis na epiderme o que difere dos achados nas lesões recentes do PFE e coincide com os achados nas lesões crônicas do PFE da era pré-corticóide. A IFD da pele lesada foi positiva para IgG em 93,75% dos casos, como ocorre nas fases iniciais do PFE, tendo sido negativa no único caso em que não houve clivagem. Adicionalmente, em oito desses doentes, foram estudados a IFD da pele sã e a imunofluorescência indireta (IFI. A IFD foi positiva em três destes casos e a IFI foi negativa nos oito.A group of 16 patients with endemic pemphigus foliaceus under corticotherapy and still showing erythematous, papulous, verrucous, in general hyperpigmented lesions, which were characterized as cortico therapy resistant lesions, were studied. Such study was made through anatomopathology and direct immunofluorescence (DIF. Anatomopathologically, such lesions showed tendencies to epitelial hyperplasia and cleavage in variable levels at the epidermis what differs from the early lesions of EPF and coincides with the chronic injuries of the EPF of the pre-corticoid era. The DIF of the injured skin was positive for IgG in 93.75% of cases, as it happens in the early stages of EPF, being negative in a simple case in which there was not cleavage. In addition, in eight of those patients, the DIF of the healthy skin and the indirect immunofluorescence (IIF were studied. The DIF was positive in three of these cases and in all eight the IIF was negative.

  16. Comparison of enzyme-linked immunosorbent assay, radioimmunoassay, complement fixation, anticomplement immunofluorescence and passive haemaglutination techniques for detecting cytomegalovirus IgG antibody

    Energy Technology Data Exchange (ETDEWEB)

    Booth, J C; Hannington, G; Bakir, T M.F.; Stern, H; Kangro, H; Griffiths, P D; Heath, R B [Saint George' s Hospital Medical School, London (UK); Saint Bartholomew' s Hospital, London (UK))

    1982-12-01

    The radioimmunoassay (RIA) and enzyme-linked immunosorbent assay (ELISA) techniques were found to be comparable in sensitivity and specificity for detecting cytomegalovirus IgG antibody, and 10 to 100 times more sensitive than complement-fixation (CF), anticomplement immunofluorescence (ACIF) and passive haemagglutination (PHA). In screening tests for antibody, the frequency of false-positive and -negative results was 0.6% for RIA and ELISA, 1.5% for CF, 1.6% for ACIF and 3.6% for PHA. PHA was the least satisfactory test, largely because of technical problems.

  17. Collider, direct and indirect detection of supersymmetric dark matter

    International Nuclear Information System (INIS)

    Baer, Howard; Park, Eun-Kyung; Tata, Xerxes

    2009-01-01

    We present an overview of supersymmetry (SUSY) searches, both at collider experiments and via searches for dark matter (DM). We focus on three DM possibilities in the SUSY context: the thermally produced neutralino, a mixture of axion and axino, and the gravitino, and compare and contrast signals that may be expected at colliders, in direct detection (DD) experiments searching of DM relics left over from the Big Bang, and indirect detection (ID) experiments designed to detect the products of DM annihilations within the solar interior or galactic halo. Detection of DM particles using multiple strategies provides complementary information that may shed light on the new physics associated with the DM sector. In contrast to the minimal supergravity (mSUGRA) model where the measured cold DM relic density restricts us to special regions mostly on the edge of the m 0 -m 1/2 plane, the entire parameter plane becomes allowed if the universality assumption is relaxed in models with just one additional parameter. Then, thermally produced neutralinos with a well-tempered mix of wino, bino and higgsino components, or with a mass adjusted so that their annihilation in the early Universe is Higgs-resonance-enhanced, can be the DM. Well-tempered neutralinos typically yield heightened rates for DD and ID experiments compared with generic predictions from mSUGRA. If instead DM consists of axinos (possibly together with axions) or gravitinos, then there exists the possibility of detection of quasi-stable next-to-lightest SUSY particles at colliding beam experiments, with especially striking consequences if the next-lightest-supersymmetric-particle (NLSP) is charged, but no DD or ID detection. The exception for mixed axion/axino DM is that DD of axions may be possible.

  18. Construction of iron-polymer-graphene nanocomposites with low nonspecific adsorption and strong quenching ability for competitive immunofluorescent detection of biomarkers in GM crops.

    Science.gov (United States)

    Yin, Kaifei; Liu, Anran; Shangguan, Li; Mi, Li; Liu, Xu; Liu, Yuanjian; Zhao, Yuewu; Li, Ying; Wei, Wei; Zhang, Yuanjian; Liu, Songqin

    2017-04-15

    We developed a new immunofluorescent biosensor by utilizing a novel nanobody (Nb) and iron-polymer-graphene nanocomposites for sensitive detection of 5-enolpyruvylshikimate-3-phosphate synthase from Agrobacdterium tumefaciens strain CP4 (CP4-EPSPS), which considered as biomarkers of genetically modified (GM) crops. Specifically, we prepared iron doped polyacrylic hydrazide modified reduced graphene nanocomposites (Fe@RGO/PAH) by in-situ polymerization approach and subsequent a one-pot reaction with hydrazine. The resulting Fe@RGO/PAH nanocomposites displayed low nonspecific adsorption to analytes (11% quenching caused by nonspecific adsorption) due to electrostatic, energetic and steric effect of the nanocomposites. After Nb immobilizing, the as-prepared Fe@RGO/PAH/Nbs showed good selectivity and high quenching ability (92% quenching) in the presence of antigen (Ag) and polyethylene glycol (PEG) modified CdTe QDs (Ag/QDs@PEG), which is a nearly 4 fold than that of the unmodified GO in same condition. The high quenching ability of Fe@RGO/PAH/Nbs can be used for detection of CP4-EPSPS based on competitive immunoassay with a linearly proportional concentration range of 5-100ng/mL and a detection limit of 0.34ng/mL. The good stability, reproducibility and specificity of the resulting immunofluorescent biosensor are demonstrated and might open a new window for investigation of fluorescent sensing with numerous multifunctional graphene based materials. Copyright © 2016 Elsevier B.V. All rights reserved.

  19. Direct and indirect detection of dissipative dark matter

    Energy Technology Data Exchange (ETDEWEB)

    Fan, JiJi; Katz, Andrey; Shelton, Jessie, E-mail: jijifan1982@gmail.com, E-mail: katz.andrey@gmail.com, E-mail: jshelton137@gmail.com [Department of Physics, Harvard University, Cambridge, MA 02138 (United States)

    2014-06-01

    We study the constraints from direct detection and solar capture on dark matter scenarios with a subdominant dissipative component. This dissipative dark matter component in general has both a symmetric and asymmetric relic abundance. Dissipative dynamics allow this subdominant dark matter component to cool, resulting in its partial or total collapse into a smaller volume inside the halo (e.g., a dark disk) as well as a reduced thermal velocity dispersion compared to that of normal cold dark matter. We first show that these features considerably relax the limits from direct detection experiments on the couplings between standard model (SM) particles and dissipative dark matter. On the other hand, indirect detection of the annihilation of the symmetric dissipative dark matter component inside the Sun sets stringent and robust constraints on the properties of the dissipative dark matter. In particular, IceCube observations force dissipative dark matter particles with mass above 50 GeV to either have a small coupling to the SM or a low local density in the solar system, or to have a nearly asymmetric relic abundance. Possible helioseismology signals associated with purely asymmetric dissipative dark matter are discussed, with no present constraints.

  20. Direct and indirect detection of dissipative dark matter

    International Nuclear Information System (INIS)

    Fan, JiJi; Katz, Andrey; Shelton, Jessie

    2014-01-01

    We study the constraints from direct detection and solar capture on dark matter scenarios with a subdominant dissipative component. This dissipative dark matter component in general has both a symmetric and asymmetric relic abundance. Dissipative dynamics allow this subdominant dark matter component to cool, resulting in its partial or total collapse into a smaller volume inside the halo (e.g., a dark disk) as well as a reduced thermal velocity dispersion compared to that of normal cold dark matter. We first show that these features considerably relax the limits from direct detection experiments on the couplings between standard model (SM) particles and dissipative dark matter. On the other hand, indirect detection of the annihilation of the symmetric dissipative dark matter component inside the Sun sets stringent and robust constraints on the properties of the dissipative dark matter. In particular, IceCube observations force dissipative dark matter particles with mass above 50 GeV to either have a small coupling to the SM or a low local density in the solar system, or to have a nearly asymmetric relic abundance. Possible helioseismology signals associated with purely asymmetric dissipative dark matter are discussed, with no present constraints

  1. Dark matter indirect detection with charged cosmic rays

    International Nuclear Information System (INIS)

    Giesen, Gaelle

    2015-01-01

    Overwhelming evidence for the existence of Dark Matter (DM), in the form of an unknown particle filling the galactic halos, originates from many observations in astrophysics and cosmology: its gravitational effects are apparent on galactic rotations, in galaxy clusters and in shaping the large scale structure of the Universe. On the other hand, a non-gravitational manifestation of its presence is yet to be unveiled. One of the most promising techniques is the one of indirect detection, aimed at identifying excesses in cosmic ray fluxes which could possibly be produced by DM annihilations or decays in the Milky Way halo. The current experimental efforts mainly focus in the GeV to TeV energy range, which is also where signals from WIMPs (Weakly Interacting Massive Particles) are expected. Focussing on charged cosmic rays, in particular antiprotons, electrons and positrons, as well as their secondary emissions, an analysis of current and foreseen cosmic ray measurements and improvements on astrophysical models are presented. Antiproton data from PAMELA imposes constraints on annihilating and decaying DM which are similar to (or even slightly stronger than) the most stringent bounds from gamma ray experiments, even when kinetic energies below 10 GeV are discarded. However, choosing different sets of astrophysical parameters, in the form of propagation models and halo profiles, allows the constraints to span over one or two orders of magnitude. In order to exploit fully the power of antiprotons to constrain or discover DM, effects which were previously perceived as sub-leading turn out to be relevant especially for the analysis of the newly released AMS-02 data. In fact, including energy losses, diffusive re-acceleration and solar modulation can somewhat modify the current bounds, even at large DM masses. A wrong interpretation of the data may arise if they are not taken into account. Finally, using the updated proton and helium fluxes just released by the AMS-02

  2. Comparative evaluations of the detection of antinuclear antibodies by means of various immunofluorescence techniques and by means of a radioimmunoassay under particular consideration of disseminated Lupus erythematodus

    International Nuclear Information System (INIS)

    Gemuend, R.

    1980-01-01

    On a group of 146 test persons (in 50 cases desseminated lupus erythematodus had been confirmed), for the first time comparative evaluations were made with four methods (A to D) under the application of a repurified fluorescinisothiocyanat FITC) serum, in order to detect antinuclear antibodies (ANA). The ANA detection was obtained by immunofluorescence (IFL) on frozen sections of mouse livers; by IFL on chicken erythrocytes smears, previously treated with hydrochloric acid; by IFL on ethanol-fixed flagellates Crithidia luciliae; and by the radioimmunoassay (RIA) of a test kit with reference sera. These two tests served to detect antibodies - with respect to negative DNA - which are of particular importance in lupous nephritis. A good correlation of both methods was proved by means of various statistic methods and by follow-up observations and examinations of the reference sera. Possible reasons responsible for the deviations, which were found between the two tests, are described. Of all 4 tests, RIA and IFL on Crithida resulted to be the most closely ones to the relevant laboratory values and reflect very evidently the activity of the desseminated lupus erethematodus. The particularly well correlation with the blood sedimentation rate, proteinuria and with the complement level becomes very obvious. The advantages and disadvantages of the applied methods are discussed and it is emphasized that at present the method of choice for the detection of DNA antibodies is the combined examination of the patient serum, both, in the IFL on Crithidia and in the RIA. (orig./MG) [de

  3. Giardia and Cryptosporidium spp. dissemination during wastewater treatment and comparative detection via immunofluorescence assay (IFA), nested polymerase chain reaction (nested PCR) and loop mediated isothermal amplification (LAMP).

    Science.gov (United States)

    Gallas-Lindemann, Carmen; Sotiriadou, Isaia; Plutzer, Judit; Noack, Michael J; Mahmoudi, Mohammad Reza; Karanis, Panagiotis

    2016-06-01

    Environmental water samples from the Lower Rhine area in Germany were investigated via immunofluorescence assays (IFAs), nested polymerase chain reaction (nested PCR) and loop-mediated isothermal amplification (LAMP) to detect the presence of Giardia spp. (n=185) and Cryptosporidium spp. (n=227). The samples were concentrated through filtration or flocculation, and oocysts were purified via centrifugation through a sucrose density gradient. For all samples, IFA was performed first, followed by DNA extraction for the nested PCR and LAMP assays. Giardia cysts were detected in 105 samples (56.8%) by IFA, 62 samples (33.5%) by nested PCR and 79 samples (42.7%) by LAMP. Cryptosporidium spp. were detected in 69 samples (30.4%) by IFA, 95 samples (41.9%) by nested PCR and 99 samples (43.6%) by LAMP. According to these results, the three detection methods are complementary for monitoring Giardia and Cryptosporidium in environmental waters. Copyright © 2016 Elsevier B.V. All rights reserved.

  4. The Muon-Induced Neutron Indirect-Detection EXperiment. MINIDEX

    Energy Technology Data Exchange (ETDEWEB)

    Palermo, Matteo

    2016-06-06

    A new experiment to measure muon-induced neutrons is introduced. The design of the Muon-Induced Neutron Indirect Detection EXperiment, MINIDEX, is presented and its installation and commissioning in the Tuebingen Shallow Underground Laboratory are described. Results from its first data taking period, run I, are presented. Muon-induced neutrons are not only an interesting physics topic by itself, but they are also an important source of background in searches for possible new rare phenomena like neutrinoless double beta decay or directly observable interactions of dark matter. These subjects are of great importance to understand the development of the early universe. Therefore, a new generation of ton-scale experiments which require extremely low background levels is under consideration. Reliable Monte Carlo simulations are needed to design such future experiments and estimate their background levels and sensitivities. The background due to muon-induced neutrons is hard to estimate, because of inconsistencies between different experimental results and discrepancies between measurements and Monte Carlo predictions. Especially for neutron production in high-Z materials, more experimental data and related simulation studies are clearly needed. MINIDEX addresses exactly this subject. Already the first five months of data taking provided valuable data on neutron production, propagation and interaction in lead. A first round of comparisons between MINIDEX data and Monte Carlo predictions are presented. In particular, the predictions of two Monte Carlo packages, based on GEANT4, are compared to the data. The data show an overall 70-100% higher rate of muon-induced events than predicted by the Monte Carlo packages. These packages also predict a faster time evolution of the muon-induced signal than observed in the data. Nevertheless, the time until the signal from the muon-induced events is completely collected was correctly predicted by the Monte Carlos. MINIDEX is foreseen

  5. A novel immunofluorescence flow cytometry technique detects the expansion of brown tides caused by Aureoumbra lagunensis to the Caribbean Sea.

    Science.gov (United States)

    Koch, F; Kang, Y; Villareal, T A; Anderson, D M; Gobler, C J

    2014-08-01

    During the past 3 decades, brown tides caused by the pelagophytes Aureococcus anophagefferens and Aureoumbra lagunensis have caused ecological and economic damage to coastal ecosystems across the globe. While blooms of A. lagunensis had previously been confined to Texas, in 2012, an expansive brown tide occurred on Florida's East Coast, causing widespread disruption within the Indian River and Mosquito Lagoons and generating renewed interest in this organism. A major impediment to detailed investigations of A. lagunensis in an ecosystem setting has been the absence of a rapid and reliable method for cell quantification. The combination of their small size (3 to 5 μm) and nondescript extracellular features makes identification and enumeration of these cells with conventional methods a challenge. Here we report the development of an immunological-based flow cytometry method that uses a fluorescently labeled antibody developed against A. lagunensis. This method is species specific, sensitive (detection limit of 1.5 × 10(3) cells ml(-1)), precise (1% relative standard deviation of replicated samples), and accurate (108% ± 8% recovery of spiked samples) over a wide range of cell concentrations. Furthermore, this method effectively quantifies A. lagunensis in both glutaraldehyde- and formalin-preserved samples, yields a high throughput of samples (∼35 samples h(-1)), and is cost-effective, making it an ideal tool for managers and scientists. This method successfully documented the recurrence of a brown tide bloom in Florida in 2013. Bloom densities were highest in June (>2.0 × 10(6) cells ml(-1)) and spanned >60 km from the Ponce de Leon inlet in the northern Mosquito Lagoon south to Titusville in the Indian River Lagoon. Low levels of A. lagunensis cells were found >250 km south of this region. This method also quickly and accurately identified A. lagunensis as the causative agent of a 2013 brown tide bloom in Guantanamo Bay, Cuba, and thus should prove useful for

  6. Prospects for indirect detection of frozen-in dark matter

    Science.gov (United States)

    Heikinheimo, Matti; Tenkanen, Tommi; Tuominen, Kimmo

    2018-03-01

    We study observational consequences arising from dark matter (DM) of nonthermal origin, produced by dark freeze-out from a hidden sector heat bath. We assume this heat bath was populated by feebly coupled mediator particles, produced via a Higgs portal interaction with the Standard Model (SM). The dark sector then attained internal equilibrium with a characteristic temperature different from the SM photon temperature. We find that even if the coupling between the DM and the SM sectors is very weak, the scenario allows for indirect observational signals. We show how the expected strength of these signals depends on the temperature of the hidden sector at DM freeze-out.

  7. Detection of Lawsonia intracellularis in formalin-fixed porcine intestinal tissue samples: comparison of immunofluorescence and in-situ hybridization, and evaluation of the effects of controlled autolysis.

    Science.gov (United States)

    Jensen, T K; Boesen, H T; Vigre, H; Boye, M

    2010-01-01

    Two methods, an immunofluorescence assay (IFA; with a Lawsonia intracellularis-specific monoclonal antibody) and fluorescent in-situ hybridization (FISH; with a specific oligonucleotide probe targeting 16S ribosomal RNA of the bacterium), were compared for their ability to detect L. intracellularis (the cause of porcine proliferative enteritis [PE]) in formalin-fixed samples of intestinal tissue. Of 69 intestinal samples with gross lesions of PE, 63 were positive by both FISH and IFA, but six were positive only by IFA. This indicated that the sensitivity of FISH was 91% that of IFA. However, both methods had a specificity of 100%. Fifty normal porcine intestines were negative by both tests. IFA was much less susceptible than FISH to the effects of autolysis. Thus, three of nine samples from pigs with PE were FISH-negative after being kept at 20 degrees C for 4 days, and seven were FISH negative after 2 weeks; after 4 weeks at this temperature, however, six of the nine samples were still IFA positive. After being kept at 4 degrees C for 12 weeks, the majority of samples (> or = 66%) were positive by both methods.

  8. Quantum dots immunofluorescence histochemical detection of EGFR gene mutations in the non-small cell lung cancers using mutation-specific antibodies

    Directory of Open Access Journals (Sweden)

    Qu YG

    2014-12-01

    Full Text Available Yan-Gang Qu,1 Qian Zhang,2 Qi Pan,3 Xian-Da Zhao,4 Yan-Hua Huang,2 Fu-Chun Chen,3 Hong-Lei Chen41Department of Pathology, The Central Hospital of Enshi Autonomous Prefecture, Enshi, 2Department of Molecular Pathology, Wuhan Nano Tumor Diagnosis Engineering Research Center, Wuhan, Hubei, People’s Republic of China; 3Department of Thoracosurgery, Traditional Chinese Medical Hospital of Wenling, Wenling, Zhejiang, People’s Republic of China; 4Department of Pathology, School of Basic Medical Science, Wuhan University, Wuhan, Hubei, People’s Republic of ChinaBackground: Epidermal growth factor receptor (EGFR mutation status plays an important role in therapeutic decision making for non-small cell lung cancer (NSCLC patients. Since EGFR mutation-specific antibodies (E746-A750del and L858R have been developed, EGFR mutation detection by immunohistochemistry (IHC is a suitable screening test. On this basis, we want to establish a new screening test, quantum dots immunofluorescence histochemistry (QDs-IHC, to assess EGFR gene mutation in NSCLC tissues, and we compared it to traditional IHC and amplification refractory mutation system (ARMS.Materials and methods: EGFR gene mutations were detected by QDs-IHC, IHC, and ADx-ARMS in 65 cases of NSCLC composed of 55 formalin-fixed, paraffin-embedded specimens and ten pleural effusion cell blocks, including 13 squamous cell carcinomas, two adenosquamous carcinomas, and 50 adenocarcinomas.Results: Positive rates of EGFR gene mutations detected by QDs-IHC, IHC, and ADx-ARMS were 40.0%, 36.9%, and 46.2%, respectively, in 65 cases of NSCLC patients. The sensitivity of QDs-IHC when detecting EGFR mutations, as compared to ADx-ARMS, was 86.7% (26/30; the specificity for both antibodies was 100.0% (26/26. IHC sensitivity was 80.0% (24/30 and the specificity was 92.31% (24/26. When detecting EGFR mutations, QDs-IHC and ADx-ARMS had perfect consistency (κ=0.882; P<0.01. Excellent agreement was observed

  9. Detection of rotator cuff lesions with indirect MR angiography

    International Nuclear Information System (INIS)

    Rudolph, J.; Lorenz, M.; Schroeder, R.; Felix, R.

    2000-01-01

    Purpose: To determine the value of indirect MR arthrography in lesions of the rotator cuff, prospectively versus arthroscopy. Methods: 63 patients with suspected shoulder pathology were examined: Oblique-coronary and axial T 1 w sequences, axial FLASH-2 D sequences, furthermore oblique-coronary T 2 - and PD-weighted sequences were taken. After intravenous administration of 0.1 mmol Gd-DTPA/kilogram body weight and active motion of the shoulder T 1 w sequences were repeated. Signal intensities (SI) inside the tendon were quantitatively measured by the ROI technique (region-of-interest) and the percentual contrast-enhancement CE was calculated. In 32 patients the results were confirmed by surgical follow-up. Results: The mean SI measured in the supraspinous tendon were higher in lesions (degeneration, impingement, partial and total rupture), before as well as after contrast medium, compared to intact findings (p [de

  10. Predicting and detecting reciprocity between indirect ecological interactions and evolution.

    Science.gov (United States)

    Estes, James A; Brashares, Justin S; Power, Mary E

    2013-05-01

    Living nature can be thought of as a tapestry, defined not only by its constituent parts but also by how these parts are woven together. The weaving of this tapestry is a metaphor for species interactions, which can be divided into three broad classes: competitive, mutualistic, and consumptive. Direct interactions link together as more complex networks, for example, the joining of consumptive interactions into food webs. Food web dynamics are driven, in turn, by changes in the abundances of web members, whose numbers or biomass respond to bottom-up (resource limitation) and top-down (consumer limitation) forcing. The relative strengths of top-down and bottom-up forcing on the abundance of a given web member depend on its ecological context, including its topological position within the food web. Top-down effects by diverse consumers are nearly ubiquitous, in many cases influencing the structure and operation of ecosystems. While the ecological effects of such interactions are well known, far less is known of their evolutionary consequences. In this essay, we describe sundry consequences of these interaction chains on species and ecosystem processes, explain several known or suspected evolutionary effects of consumer-induced interaction chains, and identify areas where reciprocity between ecology and evolution involving the indirect effects of consumer-prey interaction chains might be further explored.

  11. Validation of a commercially available indirect ELISA using a nucleocapside recombinant protein for detection of Schmallenberg virus antibodies.

    Directory of Open Access Journals (Sweden)

    Emmanuel Bréard

    Full Text Available A newly developed Enzym Like Immuno Sorbant Assay (ELISA based on the recombinant nucleocapsid protein (N of Schmallenberg virus (SBV was evaluated and validated for the detection of SBV-specific IgG antibodies in ruminant sera by three European Reference Laboratories. Validation data sets derived from sheep, goat and bovine sera collected in France and Germany (n = 1515 in 2011 and 2012 were categorized according to the results of a virus neutralization test (VNT or an indirect immuno-fluorescence assay (IFA. The specificity was evaluated with 1364 sera from sheep, goat and bovine collected in France and Belgium before 2009. Overall agreement between VNT and ELISA was 98.9% and 98.3% between VNT and IFA, indicating a very good concordance between the different techniques. Although cross-reactions with other Orthobunyavirus from the Simbu serogroup viruses might occur, it is a highly sensitive, specific and robust ELISA-test validated to detect anti-SBV antibodies. This test can be applied for SBV sero-diagnostics and disease-surveillance studies in ruminant species in Europe.

  12. Indirect Charged Particle Detection: Concepts and a Classroom Demonstration

    Science.gov (United States)

    Childs, Nicholas B.; Horányi, Mihály; Collette, Andrew

    2013-01-01

    We describe the principles of macroscopic charged particle detection in the laboratory and their connections to concepts taught in the physics classroom. Electrostatic dust accelerator systems, capable of launching charged dust grains at hypervelocities (1-100 km/s), are a critical tool for space exploration. Dust grains in space typically have…

  13. Indirect detection of dark matter with the ANTARES neutrino telescope

    International Nuclear Information System (INIS)

    Lambard, G.

    2008-01-01

    The ANTANARES telescope is composed of an array of 900 photomultipliers (12 lines) that will be immersed in the Mediterranean sea at a depth of 2500 m. The photomultipliers are sensitive to the Cherenkov light emitted by high energy muons produced in the interactions of neutrinos with matter. My work consisted in the calibration of the detector, in time and charge in order to extract the crucial data for the reconstruction of the particle tracks and the ability of the detector to distinguish the atmospheric neutrinos from astrophysical neutrinos. The first part of this work is dedicated to the today understanding of the universe and of its models and of the importance of the neutrinos as the messengers of what occurs in the remote parts of the universe. The detection of neutrinos through the Cerenkov effect is detailed and the ANTANARES detector is presented. The second part deals with the study of the background radiation due to atmospheric muons and neutrinos. A simulation is the only tool to assess the background radiation level and to be able to extract the signal due to solar neutrinos. The third part shows how the solar neutrino flux might be influenced by the interaction of dark matter with baryonic matter. A Monte-Carlo simulation has allowed us to quantify this interaction and measure its impact on the number of events detected by ANTANARES. (A.C.)

  14. Indirect photometric detection of boron cluster anions electrophoretically separated in methanol.

    Science.gov (United States)

    Vítová, Lada; Fojt, Lukáš; Vespalec, Radim

    2014-04-18

    3,5-Dinitrobenzoate and picrate are light absorbing anions pertinent to indirect photometric detection of boron cluster anions in buffered methanolic background electrolytes (BGEs). Tris(hydroxymethyl)aminomethane and morpholine have been used as buffering bases, which eliminated baseline steps, and minimized the baseline noise. In methanolic BGEs, mobilities of boron cluster anions depend on both ionic constituents of the BGE buffer. This dependence can be explained by ion pair interaction of detected anions with BGE cations, which are not bonded into ion pairs with the BGE anions. The former ion pair interaction decreases sensitivity of the indirect photometric detection. Copyright © 2014 Elsevier B.V. All rights reserved.

  15. Indirect solid-phase immunosorbent assay for detection of arenavirus antigens and antibodies

    Energy Technology Data Exchange (ETDEWEB)

    Ivanov, A P; Rezapkin, G V; Dzagurova, T K; Tkachenko, E A [Institute of Poliomyelitis anU Viral Encephalities of the U.S.S.R. Academy of Medical Sciences, Moscow

    1984-05-01

    Indirect enzyme-linked immunosorbent assay (ELISA) and solid phase radioimmunoassay (SPRIA) using either enti-human or anti-mouse IgG labelled with horseradish peroxidase and /sup 125/I, respectively, were developed for the detection of Junin, Machupo, Tacaribe, Amapari, Tamiami, Lassa and LCM arenaviruses. Both methods allow high sensitivity detection of arenavirus antigens and antibodies.

  16. Indirect fluorometric detection techniques on thin layer chromatography and effect of ultrasound on gel electrophoresis

    Energy Technology Data Exchange (ETDEWEB)

    Yinfa, Ma.

    1990-12-10

    Thin-layer chromatography (TLC) is a broadly applicable separation technique. It offers many advantages over high performance liquid chromatography (HPLC), such as easily adapted for two-dimensional separation, for whole-column'' detection and for handling multiple samples, etc. However, due to its draggy development of detection techniques comparing with HPLC, TLC has not received the attention it deserves. Therefore, exploring new detection techniques is very important to the development of TLC. It is the principal of this dissertation to present a new detection method for TLC -- indirect fluorometric detection method. This detection technique is universal sensitive, nondestructive, and simple. This will be described in detail from Sections 1 through Section 5. Section 1 and 3 describe the indirect fluorometric detection of anions and nonelectrolytes in TLC. In Section 2, a detection method for cations based on fluorescence quenching of ethidium bromide is presented. In Section 4, a simple and interesting TLC experiment is designed, three different fluorescence detection principles are used for the determination of caffeine, saccharin and sodium benzoate in beverages. A laser-based indirect fluorometric detection technique in TLC is developed in Section 5. Section 6 is totally different from Sections 1 through 5. An ultrasonic effect on the separation of DNA fragments in agarose gel electrophoresis is investigated. 262 refs.

  17. Liquid chromatographic separation and indirect detection of non-absorbing aliphatic compounds using m-nitrophenol as a detection agent

    International Nuclear Information System (INIS)

    Lee, Seung Seok; Kang, Sam Woo; Moon, Young Ja

    1991-01-01

    m-Nitrophenol(m-NP) was a detection agent for the use of the detection and separation of non-absorbing compounds such as aliphatic acids, alcohols, amines and tetraalkylammonium salts by indirect photometric detection in reversed-phase liquid chromatography. Response of samples was investigated to the several factors such as pH, temperature, and concentration of MeOH as well as concentration of detection reagent in mobile phase. The separation of several mixtures were attempted under optimum condition. (Author)

  18. The impact of the phase-space density on the indirect detection of dark matter

    International Nuclear Information System (INIS)

    Ferrer, Francesc; Hunter, Daniel R.

    2013-01-01

    We study the indirect detection of dark matter when the local dark matter velocity distribution depends upon position, as expected for the Milky Way and its dwarf spheroidal satellites, and the annihilation cross-section is not purely s-wave. Using a phase-space distribution consistent with the dark matter density profile, we present estimates of cosmic and gamma-ray fluxes from dark matter annihilations. The expectations for the indirect detection of dark matter can differ significantly from the usual calculation that assumes that the velocity of the dark matter particles follows a Maxwell-Boltzmann distribution

  19. Swelling behavior detection of irradiated U-10Zr alloy fuel using indirect neutron radiography

    Energy Technology Data Exchange (ETDEWEB)

    Sun, Yong; Huo, He-yong; Wu, Yang [Institute of Nuclear Physics and Chemistry, Chinese Academy of Engineering Physics, Mianyang (China); Key Laboratory of Neutron Physics, Chinese Academy of Engineering Physics, Mianyang (China); Li, Jiangbo [Institute of Nuclear Physics and Chemistry, Chinese Academy of Engineering Physics, Mianyang (China); Zhou, Wei; Guo, Hai-bing [Institute of Nuclear Physics and Chemistry, Chinese Academy of Engineering Physics, Mianyang (China); Key Laboratory of Neutron Physics, Chinese Academy of Engineering Physics, Mianyang (China); Li, Hang, E-mail: lihang32@gmail.com [Institute of Nuclear Physics and Chemistry, Chinese Academy of Engineering Physics, Mianyang (China); Key Laboratory of Neutron Physics, Chinese Academy of Engineering Physics, Mianyang (China); Cao, Chao; Yin, Wei; Wang, Sheng; Liu, Bin; Feng, Qi-jie; Tang, Bin [Institute of Nuclear Physics and Chemistry, Chinese Academy of Engineering Physics, Mianyang (China); Key Laboratory of Neutron Physics, Chinese Academy of Engineering Physics, Mianyang (China)

    2016-11-21

    It is hopeful that fusion-fission hybrid energy system will become an effective approach to achieve long-term sustainable development of fission energy. U-10Zr alloy (which means the mass ratio of Zr is 10%) fuel is the key material of subcritical blanket for fusion-fission hybrid energy system which the irradiation performance need to be considered. Indirect neutron radiography is used to detect the irradiated U-10Zr alloy because of the high residual dose in this paper. Different burnup samples (0.1%, 0.3%, 0.5% and 0.7%) have been tested with a special indirect neutron radiography device at CMRR (China Mianyang Research Reactor). The resolution of the device is better than 50 µm and the quantitative analysis of swelling behaviors was carried out. The results show that the swelling behaviors relate well to burnup character which can be detected accurately by indirect neutron radiography.

  20. In situ detection of the activation of Rac1 and RalA small GTPases in mouse adipocytes by immunofluorescent microscopy following in vivo and ex vivo insulin stimulation.

    Science.gov (United States)

    Takenaka, Nobuyuki; Nihata, Yuma; Ueda, Sho; Satoh, Takaya

    2017-11-01

    Rac1 has been implicated in insulin-dependent glucose uptake by mechanisms involving plasma membrane translocation of the glucose transporter GLUT4 in skeletal muscle. Although the uptake of glucose is also stimulated by insulin in adipose tissue, the role for Rac1 in adipocyte insulin signaling remains controversial. As a step to reveal the role for Rac1 in adipocytes, we aimed to establish immunofluorescent microscopy to detect the intracellular distribution of activated Rac1. The epitope-tagged Rac1-binding domain of a Rac1-specific target was utilized as a probe that specifically recognizes the activated form of Rac1. Rac1 activation in response to ex vivo and in vivo insulin stimulations in primary adipocyte culture and mouse white adipose tissue, respectively, was successfully observed by immunofluorescent microscopy. These Rac1 activations were mediated by phosphoinositide 3-kinase. Another small GTPase RalA has also been implicated in insulin-stimulated glucose uptake in skeletal muscle and adipose tissue. Similarly to Rac1, immunofluorescent microscopy using an activated RalA-specific polypeptide probe allowed us to detect intracellular distribution of insulin-activated RalA in adipocytes. These novel approaches to visualize the activation status of small GTPases in adipocytes will largely contribute to the understanding of signal transduction mechanisms particularly for insulin action. Copyright © 2017 Elsevier Inc. All rights reserved.

  1. A performance comparison of direct- and indirect-detection flat-panel imagers

    International Nuclear Information System (INIS)

    Partridge, M.; Hesse, B.-M.; Mueller, L.

    2002-01-01

    A comparison of the performance of a direct- and an indirect-detection amorphous silicon flat-panel X-ray imager is presented for a 6 MV beam. Experimental measurements of the noise characteristics, image lag, spectral response, spatial resolution and quantum efficiency are described, compared and discussed. The two systems are comprised of 512x512 pixel, 400 μm pitch, arrays of a-Si:H p-i-n photodiodes and thin-film transistors. In the direct-detection system, X-rays interact to produce electron/hole pairs directly in the silicon photodiodes. For the indirect-detection system, a phosphor screen converts energy from the incident X-rays into visible light, which is then detected by the photodiodes. Both systems are shown to be quantum noise limited, with the total electronic noise in the detector 10-15 times smaller than the Poisson noise level in detected signal. The measured lag for both systems is 1.0±0.1% or less in the first frame with subsequent signals decaying exponentially with frame read-out, with a half-life of between 3.3 and 3.8 frames. Both systems are demonstrated to have a pronounced sensitivity to low-energy multiply scattered photons, although this is shown to be effectively filtered out using a 2 mm copper build-up plate. The direct-detection system, with the 2 mm Cu build-up, shows greater sensitivity to scattered radiation than the indirect system. The spatial resolutions of both systems were effectively equal with an f 50 of 0.25 mm -1 when pixels are binned 2x2, although a slight contribution from optical scattering in the phosphor screen is seen for the indirect-detection system. The quantum efficiency of the direct-detection system is a factor of 0.45 lower than that of the indirect-detection system. The application of these detectors to megavoltage CT is discussed, with the conclusion that the indirect-detection system is to be preferred

  2. Direct and indirect fluorescent detection of tetracyclines using dually emitting carbon dots

    International Nuclear Information System (INIS)

    Qu, Fei; Sun, Zhe; Liu, Dongya; Zhao, Xianen; You, Jinmao

    2016-01-01

    The authors describe dual-emission carbon nanodots containing blue emitters (BE; peak emission at 385 nm under 315 nm excitation) and yellow emitters (YE; peak emission at 530 nm under 365 nm excitation), and how they can be applied to direct and indirect determination of tetracyclines (TCs). The direct detection scheme is based on the finding that tetracycline (TET), oxytetracycline, chlortetracycline and doxycycline quench the two emissions of the carbon dots. While direct determination is rapid and convenient, it cannot differentiate between TCs. The indirect detection scheme, in contrast, is based on the finding that Al (III) ions enhance the fluorescence of the YE in the carbon dots, and that they cause a blue shift in emission. It is, however, known that TET forms a strong complex with Al (III), and this can inhibit the interaction between Al (III) and the YE, so that the fluorescence of YE is not enhanced and blue-shifted by Al (III) in the presence of TET. This finding is exploited in a fluorescence turn-on/off assay for TET that can distinguish TET from other TCs. The linear range of indirect determination for TET extends from 1 nM to 30 μM, and the limit of detection is 0.52 nM. The indirect method was successfully applied to the determination of TET in spiked milk, fish and pork, and recoveries ranged from 91.7 to 102 %. (author)

  3. Effectiveness of direct and indirect radionuclide cystography in detecting vesicoureteral reflux

    International Nuclear Information System (INIS)

    Conway, J.J.; Kruglik, G.D.

    1976-01-01

    A modified of the direct radionuclide cystography technique to include filling, voiding, and postvoiding phases of the examination permitted a simulated comparison between direct and indirect radionuclide cystography. One hundred thirty-seven examples of reflux were documented with this technique. Of these, 96 instances of reflux (70 percent) were recorded during two or more phases and thus would have been detected by either technique. Twenty-nine examples (21 percent) were only detected during filling and thus would have been missed by the indirect radionuclide technique and by some roentgenographic techniques. Only 12 examples (9 percent) were detected during the voiding phase only. The modified direct method of radionuclide cystography, which continuously monitors the urinary tracts during filling, voiding, and postvoiding, is offered as the best current technique for assessing visicoureteral reflus

  4. Heteronuclear Correlation SSNMR Spectroscopy with Indirect Detection under Fast Magic-Angle Spinning [Book Chapter

    Energy Technology Data Exchange (ETDEWEB)

    Kobayshi, Takeshi [Ames Laboratory (AMES), Ames, IA (United States); Nishiyama, Yusuke [Ames Laboratory (AMES), Ames, IA (United States); Pruski, Marek [Ames Laboratory (AMES), Ames, IA (United States)

    2018-01-01

    The main focus of this chapter is to address experimental strategies on the subject by providing a hands-on guide to fast MAS experiments, with a particular focus on indirect detection. Although our experience is limited to our respective laboratories in Ames and Yokohama, we hope that our descriptions of experimental setups and optimization procedures are sufficiently general to be applicable to all modern instruments. The chapter is organized as follows. Section 2 below introduces briefly the fast MAS technology and its main advantages. In Section 3, we describe the hardware associated with this remarkable technology and provide practical advices on its use, including procedures for loading and unloading the samples, maintaining the probe, reducing t1 noise, etc. In Section 4, we describe the principles and hands-on aspects of experiments involving the indirect detection of spin-1/2 and 14N nuclei

  5. Design of indirect solid-phase immunosorbent methods for detecting arenavirus antigens and antibodies

    Energy Technology Data Exchange (ETDEWEB)

    Ivanov, A P; Rezapkin, G V; Dzagurova, T K; Tkachenko, E A

    1984-05-01

    Specifications have been elaborated for formulating indirect solid-phase enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay (SPRIA) methods that employ anti-human and anti-mice G class immunoglobulin (IgG), conjugated with horseradish peroxidase and /sup 125/I for detecting the arenaviruses Junin, Machupo, Tacaribe, Amalpari, Tamiami, Lassa, and LCM (lymphocytic choriomeningitis). These methods make it possible to identify with a high degree of sensitivity arenavirus antigens and antibodies in various kinds of material.

  6. A performance comparison of direct- and indirect-detection flat-panel imagers

    CERN Document Server

    Partridge, M; Müller, L

    2002-01-01

    A comparison of the performance of a direct- and an indirect-detection amorphous silicon flat-panel X-ray imager is presented for a 6 MV beam. Experimental measurements of the noise characteristics, image lag, spectral response, spatial resolution and quantum efficiency are described, compared and discussed. The two systems are comprised of 512x512 pixel, 400 mu m pitch, arrays of a-Si:H p-i-n photodiodes and thin-film transistors. In the direct-detection system, X-rays interact to produce electron/hole pairs directly in the silicon photodiodes. For the indirect-detection system, a phosphor screen converts energy from the incident X-rays into visible light, which is then detected by the photodiodes. Both systems are shown to be quantum noise limited, with the total electronic noise in the detector 10-15 times smaller than the Poisson noise level in detected signal. The measured lag for both systems is 1.0+-0.1% or less in the first frame with subsequent signals decaying exponentially with frame read-out, with...

  7. Detection of antibodies in human serum using trimellityl-erythrocytes: direct and indirect haemagglutination and haemolysis.

    Science.gov (United States)

    Turner, E S; Pruzansky, J J; Patterson, R; Zeiss, C R; Roberts, M

    1980-02-01

    Utilizing trimellityl-erythrocytes (TM-E), antibodies were detected in sera of seven workers with trimellitic anhydride (TMA) induced airway syndromes by direct haemagglutination, indirect haemagglutination with anti-human IgG, IgA or IgM or by haemolysis. Detectable levels of antibody were obtained with all three methods. The most sensitive technique was indirect haemagglutination using anti-IgG. When added as an inhibitor, TM-human serum albumin produced a 10- to 800-fold reduction in titres. TM-ovalbumin of similar epitope density was less inhibitory and sodium trimellitate the least inhibitory on a molar basis. All of the assays using haptenized human red cells were also capable of detecting anti-TM antibodies in Rhesus monkeys whose airways had been exposed to TMA. These assays are useful for detecting anti-TM antibodies and may also be adapted to demonstrate antibodies induced against other inhaled haptens in sera of environmentally exposed individuals or in animal models of such exposure.

  8. Role of electroweak radiation in predictions for dark matter indirect detection

    Energy Technology Data Exchange (ETDEWEB)

    Ali Cavasonza, Leila; Pellen, Mathieu; Kraemer, Michael [RWTH Aachen, Aachen (Germany)

    2015-07-01

    A very exciting challenge in particle and astroparticle physics is the exploration of the nature of dark matter. The evidences of the existence of dark matter are also the strongest phenomenological indications for physics beyond the Standard Model. A huge experimental effort is currently made at colliders and via astrophysical experiments to shed light on the nature of dark matter: dark matter may be produced at colliders or detected through direct and indirect detection experiments. The interplay and complementarity between these different approaches offers extraordinary opportunities to improve our understanding of the nature of dark matter or to set constraints on dark matter models. In indirect detection one searches for dark matter annihilation products, that produce secondary antimatter particles like positrons and antiprotons. Such antimatter particles propagate through the Galaxy and can be detected at Earth by astrophysical experiments. Particularly interesting is the importance of electroweak corrections to the predictions for the expected fluxes at Earth. The inclusion of EW radiation from the primary dark matter annihilation products can significantly affect the spectra of the secondary SM particles. The EW radiation can be described using fragmentation functions, as done for instance in QCD. We study the quality of this approximation in a simplified SUSY model and in a UED model.

  9. Indirect detection of neutralino dark matter up to TeV scale

    International Nuclear Information System (INIS)

    Hooper, Dan

    2001-01-01

    In this paper, we will describe the results of SUSY parameter space searches including minimal supergravity, non-universal supergravity and minimal supersymmetry and the implications on the indirect detection of neutralino dark matter. We give special attention to the effects of detector thresholds, solar absorption of neutrinos and hadronization of neutralino annihilation products. These effects are known to be important in calculating accurate event rates [1]. We chose also to focus on models which predict a heavy lightest neutralino (several hundred GeV to several TeV). These models have been selected for several reasons including their inaccessibility in future collider searches

  10. Improved detection of canine Angiostrongylus vasorum infection using real-time PCR and indirect ELISA.

    Science.gov (United States)

    Jefferies, Ryan; Morgan, Eric R; Helm, Jenny; Robinson, Matthew; Shaw, Susan E

    2011-12-01

    This study reports the development of a real-time PCR assay and an indirect ELISA to improve on current detection of canine Angiostrongylus vasorum infection. A highly specific fluorescent probe-based, real-time PCR assay was developed to target the A. vasorum second internal transcribed spacer region and detected DNA in EDTA blood, lung tissue, broncho-alveolar larvage fluid, endotracheal mucus, pharyngeal swabs and faecal samples. PCR was fast (∼1 h), highly efficient when using EDTA blood samples, consistently detected a single molecule of parasite DNA and did not amplify DNA from other parasitic nematodes or definitive host species. An indirect ELISA was also developed using the soluble protein fraction from adult A. vasorum worms. Some cross-reactive antigen recognition was observed when tested against sera from dogs infected with Crenosoma vulpis (n = 8), Toxocara canis (n = 5) and Dirofilaria immitis (n = 5). This was largely overcome by setting the cut-off for a positive result at an appropriately high level. Field evaluation of the real-time PCR and ELISA was conducted by testing sera and EDTA blood from dogs with suspected A. vasorum infection (n = 148) and compared with the Baermann's larval migration test in faeces. Thirty-one dogs were positive by at least one test. Of these, 20 (65%) were detected by the Baermann method, 18 (58%) by blood PCR, 24 (77%) by ELISA and 28 (90%) by blood PCR and ELISA together. Combined testing using real-time PCR and ELISA therefore improved the detection rate of A. vasorum infection and holds promise for improved clinical diagnosis and epidemiological investigation.

  11. Towards understanding thermal history of the Universe through direct and indirect detection of dark matter

    Energy Technology Data Exchange (ETDEWEB)

    Roszkowski, Leszek; Trojanowski, Sebastian [National Centre for Nuclear Research, Hoża 69, 00-681 Warsaw (Poland); Turzyński, Krzysztof, E-mail: leszek.roszkowski@ncbj.gov.pl, E-mail: sebastian.trojanowski@uci.edu, E-mail: Krzysztof-Jan.Turzynski@fuw.edu.pl [Institute of Theoretical Physics, Faculty of Physics, University of Warsaw, ul. Pasteura 5, 02-093 Warsaw (Poland)

    2017-10-01

    We examine the question to what extent prospective detection of dark matter by direct and indirect- detection experiments could shed light on what fraction of dark matter was generated thermally via the freeze-out process in the early Universe. By simulating putative signals that could be seen in the near future and using them to reconstruct WIMP dark matter properties, we show that, in a model- independent approach this could only be achieved in a thin sliver of the parameter space. However, with additional theoretical input the hypothesis about the thermal freeze-out as the dominant mechanism for generating dark matter can potentially be verified. We illustrate this with two examples: an effective field theory of dark matter with a vector messenger and a higgsino or wino dark matter within the MSSM.

  12. Should varicella-zoster virus culture be eliminated? A comparison of direct immunofluorescence antigen detection, culture, and PCR, with a historical review.

    Science.gov (United States)

    Wilson, D A; Yen-Lieberman, B; Schindler, S; Asamoto, K; Schold, J D; Procop, G W

    2012-12-01

    A comparison of direct fluorescent-antibody assay (DFA), culture, and two PCR assays disclosed sensitivities of 87.8%, 46.3%, and 97.6% and 100%, respectively. We reviewed 1,150 results for clinical specimens submitted for DFA and culture and found that only 17 were culture positive/DFA negative. The incremental cost to detect these 17 positives was $3,078/specimen.

  13. Immunofluorescence in multiple tissues utilizing serum from a patient affected by systemic lupus erythematosus

    Directory of Open Access Journals (Sweden)

    Piotr Brzezinski

    2012-01-01

    Full Text Available Introduction: Lupus erythematosus is a chronic, inflammatory autoimmune disease that can affect multiple organs. Lupus can affect many parts of the body, especially in systemic lupus erythematosus (SLE; affected tissues may include the joints, skin, kidneys, heart, lungs, blood vessels, and brain. Case report: A 46-year-old female presented with pruritus, photosensitivity and edema of the cheeks of about 2 years duration, and was evaluated by a dermatologist. On examination, multiple telangiectasias were present on the cheeks, with erythema, edema and a malar rash observed. A review of systems documented breathing difficulty and pleuitic pain, joint pain and joint edema, photosensitivity, cardiac dysrhythmia, and periodic pain in the back close to the kidneys. Methods: Skin biopsies for hematoxylin and eosin testing, as well for direct and indirect immunofluorescence were performed, in addition to multiple diagnostic blood tests, chest radiography and directed immunologic testing. Results: The blood testing showed elevated C-reactive protein. Direct and indirect immunofluorescence testing utilizing monkey esophagus, mouse and pig heart and kidney, normal human eyelid skin and veal brain demonstrated strong reactivity to several components of smooth muscle, nerves, blood vessels, skin basement membrane zone and sweat gland ducts and skin meibomian glands. Anti-endomysium antibodies were detected as well as others, especially using FITC conjugated Complement/C1q, FITC conjugated anti-human immunoglobulin IgG and FITC conjugated anti-human fibrinogen. Conclusions: We conclude that both direct and indirect immunofluorescence using several substrates can unveil previously undocumented autoantibodies in multiple organs in lupus erythematosus, and that these findings could be utilized to complement existing diagnostic testing for this disorder.

  14. Evaluation and Comparison of Enzyme Immunoassay (Eia and Acid Fast Staining with Confirmation by Immunofluorescent Antibody Assay for Detection of Cryptosporidium Species in Infants and Young Children.

    Directory of Open Access Journals (Sweden)

    D Dorostcar Moghaddam

    2005-01-01

    Full Text Available Introduction: Cryptosporidiosis is prevalent world wide, causing a variety of problems ranging from acute, self-limiting diarrhea to fatal cases in immunocompromised persons, particulary those with acquired immunodeficiency (AIDS. Diagnosis of Cryptosporidium is made by identification of oocysts in stool specimens. The detection is most commonly made by the acid-fast staining method followed by microscopic examination which has low specificity and sensitivity. Material and Methods: In the present study, we evaluated diagnostic utility of a commercially available enzyme immunoassay (EIA, which detects Cryptosporidium-Specific antigen (CSA in 204 unprocessed stool specimens obtained from patients less than 3 years of age. Results: When compared with the routine screening procedure applied in this field study (screening by acid-fast staining and microscopy after concentration of positive results by IFA, both sensitivity and specificity were 98%. Of the 139 specimens negative by microscopy, 13 (9.3% were positive by EIA, 11 of which were confirmed by inhibition with antibody to Cryptosporidia-specific antigen. Conclusion: The EIA is an important tool for identifying Cryptosporidium in fecal specimens in field studies since it is sensitive, specific, simple to use and unaffected by the presence of a preservative.

  15. Indirectly detected chemical shift correlation NMR spectroscopy in solids under fast magic angle spinning

    Energy Technology Data Exchange (ETDEWEB)

    Mao, Kanmi [Iowa State Univ., Ames, IA (United States)

    2011-01-01

    The development of fast magic angle spinning (MAS) opened up an opportunity for the indirect detection of insensitive low-γ nuclei (e.g., 13C and 15N) via the sensitive high-{gamma} nuclei (e.g., 1H and 19F) in solid-state NMR, with advanced sensitivity and resolution. In this thesis, new methodology utilizing fast MAS is presented, including through-bond indirectly detected heteronuclear correlation (HETCOR) spectroscopy, which is assisted by multiple RF pulse sequences for 1H-1H homonuclear decoupling. Also presented is a simple new strategy for optimization of 1H-1H homonuclear decoupling. As applications, various classes of materials, such as catalytic nanoscale materials, biomolecules, and organic complexes, are studied by combining indirect detection and other one-dimensional (1D) and two-dimensional (2D) NMR techniques. Indirectly detected through-bond HETCOR spectroscopy utilizing refocused INEPT (INEPTR) mixing was developed under fast MAS (Chapter 2). The time performance of this approach in 1H detected 2D 1H{l_brace}13C{r_brace} spectra was significantly improved, by a factor of almost 10, compared to the traditional 13C detected experiments, as demonstrated by measuring naturally abundant organic-inorganic mesoporous hybrid materials. The through-bond scheme was demonstrated as a new analytical tool, which provides complementary structural information in solid-state systems in addition to through-space correlation. To further benefit the sensitivity of the INEPT transfer in rigid solids, the combined rotation and multiple-pulse spectroscopy (CRAMPS) was implemented for homonuclear 1H decoupling under fast MAS (Chapter 3). Several decoupling schemes (PMLG5m$\\bar{x}$, PMLG5mm$\\bar{x}$x and SAM3) were analyzed to maximize the performance of through-bond transfer based

  16. Evaluation of four indirect ELISA systems for the detection of trypanosomal antibodies in bovine serum

    International Nuclear Information System (INIS)

    Ndamkou, C.N.; Yomo, J.P.

    2000-01-01

    Four indirect-ELISA systems developed by the Joint FAO/IAEA Division for the detection of trypanosomal antibodies in bovine serum were evaluated in the field. Internal quality control data obtained were good showing that pre-coating plates with antigen increase the robustness of the assay and contribute to its standardisation. ELISA systems derived from Trypanosoma vivax antigen lysates gave a better performance than ELISA systems using T. congolense antigens. Sensitivity and specificity corresponding to the highest accuracy were 86-87% and 83-85% respectively. When comparing the two ELISA systems utilising T. vivax antigens, there was no significant difference between native and denatured antigens and diagnostic threshold was higher for denatured antigens. (author)

  17. Evaluation of an indirect ELISA for detection and typing of foot-and-mouth disease virus

    International Nuclear Information System (INIS)

    Prado, J.A.

    1998-01-01

    An indirect enzyme linked immunosorbent assay (ELISA) kit was used for diagnosis of foot-and-mouth disease virus (FMDV) types O1, A23, C3 which occurred in Rio Grande do Sul State, Southern Brazil during 1984-1994. The samples were randomly selected and tested by ELISA, Complement Fixation Test (CFT) and in tissue culture. Out of 106 samples 78 (73,5%) were positive by ELISA and 39 (36,8%) were found positive in CFT, when original suspensions were used. Once these samples were inoculated onto tissue culture both tests gave similar results, although ELISA picked up more positive samples during the 1st passage in tissue culture. The negative samples (16) included in this study were negative in all tests. The ELISA was more sensitive than and as specific as CFT. ELISA and tissue culture together were shown to be a better system for detection of foot-and-mouth disease virus antigen than CFT. (author)

  18. Mixed Wino Dark Matter: consequences for direct, indirect and collider detection

    International Nuclear Information System (INIS)

    Baer, Howard; Mustafayev, Azar; Park, Eun-Kyung; Profumo, Stefano

    2005-01-01

    In supersymmetric models with gravity-mediated SUSY breaking and gaugino mass unification, the predicted relic abundance of neutralinos usually exceeds the strict limits imposed by the WMAP collaboration. One way to obtain the correct relic abundance is to abandon gaugino mass universality and allow a mixed wino-bino lightest SUSY particle (LSP). The enhanced annihilation and scattering cross sections of mixed wino dark matter (MWDM) compared to bino dark matter lead to enhanced rates for direct dark matter detection, as well as for indirect detection at neutrino telescopes and for detection of dark matter annihilation products in the galactic halo. For collider experiments, MWDM leads to a reduced but significant mass gap between the lightest neutralinos so that Z-tilde 2 two-body decay modes are usually closed. This means that dilepton mass edges- the starting point for cascade decay reconstruction at the CERN LHC- should be accessible over almost all of parameter space. Measurement of the m Z-tilde2 -m Z-tilde1 mass gap at LHC plus various sparticle masses and cross sections as a function of beam polarization at the International Linear Collider (ILC) would pinpoint MWDM as the dominant component of dark matter in the universe

  19. Facilitating the indirect detection of genomic DNA in an electrochemical DNA biosensor using magnetic nanoparticles and DNA ligase

    Directory of Open Access Journals (Sweden)

    Roozbeh Hushiarian

    2015-12-01

    This technique was found to be reliably repeatable. The indirect detection of genomic DNA using this method is significantly improved and showed high efficiency in small amounts of samples with the detection limit of 5.37 × 10−14 M.

  20. Validation of an indirect ELISA for the detection of Trypanosoma congolense antibodies in Ethiopian cattle

    International Nuclear Information System (INIS)

    Tadesse, Y.; Kefyalew, H.; Kembata, G.; Nega, A.

    2000-01-01

    Control and eradication of African Animal Trypanosomosis can be achieved if the reliability of the methods to diagnose the disease could be improved. The techniques currently used in the diagnosis of trypanosomosis in Ethiopia are not sufficiently sensitive to detect all infected animals. In order to improve disease diagnosis the indirect antibody detection ELISA, developed by FAO/IAEA, was evaluated under field conditions. Accordingly, reference serum samples were collected from trypanosomosis free and endemic areas. Serum samples negative for trypanosomes were collected from the Central highlands of Ethiopia where there is no previous record of trypanosomosis. The samples were used to establish the threshold and the specificity of the test. Trypanosoma congolense positive sera (based on thin and thick smears and BCT) were collected from endemic areas in the Southwestern part of the country to estimate the sensitivity of the test. Out of 701 negative serum samples, 690 were identified as negative with the indirect antibody ELISA, whereas the remaining 11 were detected as positive. Moreover, of the 282 infected samples the ELISA detected 155 sera as positive, but the remaining 127 cases fell in the negative range. The positive/negative threshold established from negative reference sera was found to be 81.38%. Based on this threshold the specificity of the test was 98.43%, whilst the sensitivity was calculated as 54.96%. Thus, the complementary use of both the ELISA and parasitological methods is encouraged. Since the internal quality controls (IQC) did not fall in the ranges prescribed in the protocol provided by FAO/IAEA precision was achieved by comparing the plate to plate variation of the IQC based on the means plot. Accordingly the assay process indicated that there was no significant difference between individual mean of each plate regarding the strong positive (C++) and moderate positive (C+) controls. Nevertheless, considerable discrepancies were

  1. Comparison of indirect immunofluorescence test for measles antibodies with haemagglutination inhibition and plaque neutralization tests Comparação da reação de imunofluorescência indireta para o vírus do sarampo com as reações de inibição da hemaglutinação e neutralização por redução de placas

    Directory of Open Access Journals (Sweden)

    Vanda Akico Ueda Fick de Souza

    1990-10-01

    Full Text Available Indirect Immunofluorescence (IFA, Plaque Reduction Neutralization (PRN and Haemagglutination Inhibition (HI tests for measles antibodies were carried out in 197 sera obtained from umbilical cord and vaccinated children. The IFA was also applied to blood samples collected with filter paper. IFA results demonstrated that the test is relatively simple to perform, with good reproducibility for different antigen lots. Good correlation was obtained between IFA, PRN and HI antibody titers. Better correlation was demonstrated with IFA and PRN than with HI and PRN tests. Sensitivity of IFA in detecting antibody was less effective than PRN, however more effective than HI using rhesus monkey red blood cells. PRN antibody titers over 100 were detected by IFA but not by HI (9.7% with negative results. IFA may be of considerable practical use and able to substitute HI in Seroepidemiological surveys and to evaluate vaccine efficacy. It also can be simplified by employing filter paper collected samples.As reações de imunofluorescência indireta (RIF, neutralização por redução de placas (RNP e inibição da hemaglutinação (RIH para detecção de anticorpos para o vírus do sarampo foram aplicadas a 197 soros provenientes de cordão umbilical e de crianças vacinadas contra o sarampo. Avaliou-se ainda a aplicação da RIF em amostras colhidas em papel de filtro. A RIF apresentou-se como uma prova de execução relativamente simples, de boa reprodutibilidade com diferentes partidas de antígeno. Observou-se boa correlação entre os títulos de anticorpos obtidos por RIF, RNP e RIH. Com a RNP, a RIF apresentou maior correlação que a RIH. A sensibilidade da RIF na detecção de anticorpos contra o sarampo foi menor que RNP, porém superior à da RIH com hemácias de macaco rhesus. Anticorpos com títulos superiores a 100 pela RNP foram sistematicamente detectados pela RIF, mas não por RIH (9,7% de resultados negativos. A RIF pode ser de grande utilidade

  2. Establishment of an indirect ELISA for detection of the novel antifibrotic peptide M10.

    Directory of Open Access Journals (Sweden)

    Tanjina Akter

    Full Text Available M10 is a ten amino acid peptide generated from the intracellular cytoplasmic tail of the hepatocyte growth factor (HGF receptor c-Met following cleavage by caspase-3. Recently we reported that M10 interacts with Smad2 and demonstrates antifibrotic properties in vitro and in vivo and can be advanced into a novel antifibrotic remedy. The current study was undertaken to develop an immunoassay to measure M10 concentration in biological specimens.An Indirect Enzyme-Linked Immunosorbent Assay (ELISA for detection of M10 in biological fluids was developed using pharmaceutical grade synthetic M10 as a calibrator and commercially available anti-c-Met C12 antibody.M10 ELISA specifically detected in plasma M10, but not a scrambled peptide, following a single intraperitoneal administration of M10 (1mg/kg to mice. The detection limit was 9.6 ng/ml, and the measuring limit was between 15 ng/ml and 200 ng/ml. The recovery limits of M10 were between 80% and 120%; intra-assay coefficient of variation was between 5.3% and 6.3%; inter-assay coefficient of variation was between 5.0% and 8.0% over the buffer concentration tested in the range from 15 ng /ml to 250 ng /ml. The peak of M10 concentration following a single intraperitoneal injection (1mg/kg was achieved within 6 hours and declined to minimal levels by 48 hours. The experimentally obtained half-life for M10 was comparable to the theoretically predicted half-life for M10.We have established a highly sensitive ELISA to detect the antifibrotic peptide M10 in plasma samples, which should prove to be a novel tool to study the pharmacokinetics and efficacy of M10 in the treatment of fibroproliferative disorders.

  3. Direct immunofluorescence for the diagnosis of legionellosis

    Directory of Open Access Journals (Sweden)

    David JM Haldane

    1993-01-01

    Full Text Available Culture and direct immunofluorescent microscopy (DFA results for Legionella pneumophila were reviewed over a two-year period. In the first year, a positive result was defined as having at least one morphologically typical fluorescing organism. In the second year, a positive was defined as at least five typical fluorescing organisms. Despite these stricter criteria and other measures to reduce the possibility of reagent contamination, there was no statistically significant difference in the sensitivity or specificity of the DFA in the two years for sputa, deep specimens or overall. Of 37 sputum specimens from infected patients, 16 were positive on DFA. Thirty-two of 38 positive patients were detected by sputum culture. DFA can provide rapid diagnostic information but cannot be used to rule out the diagnosis. Sputum is a useful specimen for the initial laboratory investigation of patients with legionellosis.

  4. Doppler effect on indirect detection of dark matter using dark matter only simulations

    Science.gov (United States)

    Powell, Devon; Laha, Ranjan; Ng, Kenny C. Y.; Abel, Tom

    2017-03-01

    Indirect detection of dark matter is a major avenue for discovery. However, baryonic backgrounds are diverse enough to mimic many possible signatures of dark matter. In this work, we study the newly proposed technique of dark matter velocity spectroscopy [E. G. Speckhard, K. C. Y. Ng, J. F. Beacom, and R. Laha, Phys. Rev. Lett. 116, 031301 (2016), 10.1103/PhysRevLett.116.031301]. The nonrotating dark matter halo and the Solar motion produce a distinct longitudinal dependence of the signal which is opposite in direction to that produced by baryons. Using collisionless dark matter only simulations of Milky Way like halos, we show that this new signature is robust and holds great promise. We develop mock observations by a high energy resolution x-ray spectrometer on a sounding rocket, the Micro-X experiment, to our test case, the 3.5 keV line. We show that by using six different pointings, Micro-X can exclude a constant line energy over various longitudes at ≥3 σ . The halo triaxiality is an important effect, and it will typically reduce the significance of this signal. We emphasize that this new smoking gun in motion signature of dark matter is general and is applicable to any dark matter candidate which produces a sharp photon feature in annihilation or decay.

  5. Multi-Layer Organic Squaraine-Based Photodiode for Indirect X-Ray Detection

    Science.gov (United States)

    Iacchetti, Antonio; Binda, Maddalena; Natali, Dario; Giussani, Mattia; Beverina, Luca; Fiorini, Carlo; Peloso, Roberta; Sampietro, Marco

    2012-10-01

    The paper presents an organic-based photodiode coupled to a CsI(Tl) scintillator to realize an X-ray detector. A suitable blend of an indolic squaraine derivative and of fullerene derivative has been used for the photodiode, thus allowing external quantum efficiency in excess of 10% at a wavelength of 570 nm, well matching the scintillator output spectrum. Thanks to the additional deposition of a 15 nm thin layer of a suitable low electron affinity polymer, carriers injection from the metal into the organic semiconductor has been suppressed, and dark current density as low as has been obtained, which is comparable to standard Si-based photodiodes. By using a collimated X-ray beam impinging onto the scintillator mounted over the photodiode we have been able to measure current variations in the order of 150 pA on a dark current floor of less than 50 pA when operating the X-ray tube in switching mode, thus proving the feasibility of indirect X-ray detection by means of organic semiconductors.

  6. AMS-02 positron excess and indirect detection of three-body decaying dark matter

    Energy Technology Data Exchange (ETDEWEB)

    Cheng, Hsin-Chia [Department of Physics, University of California, Davis, CA 95616 (United States); Huang, Wei-Chih [Fakultät für Physik, Technische Universität Dortmund, 44221 Dortmund (Germany); Huang, Xiaoyuan [Physik-Department T30d, Technische Universität München, James-Franck-Straße, D-85748 Garching (Germany); Low, Ian [High Energy Physics Division, Argonne National Laboratory, Argonne, IL 60439 (United States); Tsai, Yue-Lin Sming [Kavli IPMU (WPI), University of Tokyo, Kashiwa, Chiba 277-8583 (Japan); Yuan, Qiang, E-mail: cheng@physics.ucdavis.edu, E-mail: wei-chih.huang@tu-dortmund.de, E-mail: huangxiaoyuan@gmail.com, E-mail: ilow@northwestern.edu, E-mail: smingtsai@gmail.com, E-mail: yuanq@pmo.ac.cn [Key Laboratory of Dark Matter and Space Astronomy, Purple Mountain Observatory, Chinese Academy of Sciences, Nanjing 210008 (China)

    2017-03-01

    We consider indirect detection of meta-stable dark matter particles decaying into a stable neutral particle and a pair of standard model fermions. Due to the softer energy spectra from the three-body decay, such models could potentially explain the AMS-02 positron excess without being constrained by the Fermi-LAT gamma-ray data and the cosmic ray anti-proton measurements. We scrutinize over different final state fermions, paying special attention to handling of the cosmic ray background and including various contributions from cosmic ray propagation with the help of the LIKEDM package. It is found that primary decays into an electron-positron pair and a stable neutral particle could give rise to the AMS-02 positron excess and, at the same time, stay unscathed against the gamma-ray and anti-proton constraints. Decays to a muon pair or a mixed flavor electron-muon pair may also be viable depending on the propagation models. Decays to all other standard model fermions are severely disfavored.

  7. Avaliação do teste de imunofluorescência indireta para diagnóstico da filariose bancroftiana usando a microfilária de W. bancrofti como antígeno, em Recife-PE, Brasil Evaluation of indirect immunofluorescence test for bancroftian filariasis using Wuchereria bancroft microfilariae as the antigen in Recife, Brazil

    Directory of Open Access Journals (Sweden)

    Gerusa Dreyer

    1991-10-01

    Full Text Available Analisou-se o teste de imunofluorescência indireta com microfilárias de W. bancrofti tratadas pela papaína, como antígeno, amplamente utilizado em Recife para o imunodiagnóstico da filariose linfática. Foram testados soros de 50 pacientes portadores das diversas formas clínicas da doença, incluindo microfilaremia assintomática, eosinofilia pulmonar tropical, elefantíase de membros inferiores, linfagite aguda e quilúria. Para o grupo controle, foram selecionados 50 indivíduos vivendo pelo menos há 5 anos em área endêmica, sem nenhuma evidencia clínica e/ou laboratorial da doença, constituindo os chamados endêmicos normais. A sensibilidade e especificidade do teste, segundo diferentes pontos de corte, mostraram a impossibilidade de diferenciação entre o grupo controle e o grupo sabidamente infectado. Também não foi possível estabelecer correlação entre os títulos encontrados e as diferentes formas clínicas. Foi considerada a existência de reações cruzadas relacionadas a helmintíases intestinais, porém nenhuma relação direta foi encontrada.The authors analysed the indirect immunofluorescence assay, for the diagnosis of bancroftian filariasis using papain treated W. bancrofti microfilariae as antigen, widely used in Recife - Brazil. Sera from 50 patients with several clinical forms of the disease including asymptomatic carriers, tropical pulmonary eosinophilia, elephantiasis, filarial fever and chyluria were analysed. For the control group, 50 individuals were selected, living at least 5 years in endemic area, with neither previous DEC treatment nor clinical-laboratory evidences of the disease, called normals endemic. The sensitivity and specificity were analised taking into account different cut off values. It was not possible to differentiate infected individuals from the control group. It was not even possible to establish any correlation with IMF titers among different clinical presentation of the disease

  8. Development of flow injection method for indirect copper determination with amperometric detection in drinking water samples

    Directory of Open Access Journals (Sweden)

    Nikolić-Mandić Snežana

    2012-01-01

    Full Text Available A gas-diffusion flow injection method with amperometric detection for indirect copper determination on a silver electrode is developed. The flow through system is equipped with two injection valves and a gas-diffusion unit. In the first step, a signal of cyanide solution was recorded. In the following step a signal of cyanide in the presence of copper was measured. Interferences (Cd(II, Co(II, Ag(I, Ni(II, Fe(III, Hg(II and Zn(II were investigated and successfully removed. The calibration graph is linear in the range 1-90 μmol dm-3 of copper, correlation coefficient is 0.993, the regression equation is I = (0.0455±0.0015c + (0.4611±0.0671, I is relative signal decrease in μA and c is concentration in μmol dm-3. Relative standard deviation for six consecutive injections of 30 μmol dm-3 copper(II was 1.47 % and for 1 μmol dm-3 copper(II was 3.40 %. The detection limit, calculated as 3 s/m (where s is a standard deviation of nine measurement of a reagent blank and m is the slope of the calibration curve, was 0.32 μmol dm-3, which corresponds to 2.44 ng of copper(II (loop volume was 0.12 cm3. The method enables 60 analyses per hour and it was successfully applied on determination of copper in drinking water samples. [Acknowledgements. The authors acknowledge the grant from the Ministry of Education and Science of the Republic of Serbia, Project number 172051

  9. Detection of hidden hazelnut protein in food by IgY-based indirect competitive enzyme-immunoassay

    NARCIS (Netherlands)

    Baumgartner, S.; Bremer, M.G.E.G.; Kemmers - Voncken, A.E.M.; Smits, N.G.E.; Haasnoot, W.; Banks, J.; Reece, P.; Danks, C.; Tomkies, V.; Immer, U.; Schmitt, K.; Krska, R.

    2004-01-01

    The development of an indirect competitive enzyme-immunoassay for the detection of hidden hazelnut protein in complex food matrices is described. A sensitive and selective polyclonal antibody was raised by immunisation of laying hens with protein extracts from roasted hazelnuts. In contrast to

  10. Indirect photometric detection of boron cluster anions electrophoretically separated in methanol

    Czech Academy of Sciences Publication Activity Database

    Vítová, Lada; Fojt, Lukáš; Vespalec, Radim

    2014-01-01

    Roč. 1338, APR 2014 (2014), s. 174-183 ISSN 0021-9673 Institutional support: RVO:68081707 Keywords : Boron cluster * Capillary electrophoresis * Indirect photometry Subject RIV: BO - Biophysics Impact factor: 4.169, year: 2014

  11. An effective model for fermion dark matter. Indirect detection of supersymmetric dark matter in astronomy with the CELESTE Telescope; Modele effectif de matiere noire fermionique. Detection indirecte de matiere noir supersymetrique en astronomie avec le Telescope CELESTE

    Energy Technology Data Exchange (ETDEWEB)

    Lavalle, Julien [Ecole Doctorale des Sciences Fondamentales, Universite Blaise Pascal, Aubiere (France)

    2004-07-01

    The purpose of this thesis is to discuss both phenomenological and experimental aspects of Dark Matter, related to its indirect detection with gamma-ray astronomy. In the MSSM framework, neutralinos arise as natural candidates to non-baryonic and Cold Dark Matter, whose gravitational effects manifest in the Universe at different scales. As they are Majorana particles, they may in principle annihilate in high density regions, as the centres of galaxies, and produce gamma rays. Nevertheless, the expected fluxes are basically low compared to experimental sensitivities. After estimating gamma fluxes from M31 and Draco galaxies in the MSSM scheme, we first generalize the MSSM couplings by studying an effective Lagrangian. We show that the only constraint of imposing a relic abundance compatible with recent measurements obviously deplete significantly the gamma ray production, but also that predictions in this effective approach are more optimistic for indirect detection than the MSSM. In a second part, we present the indirect searches for Dark Matter performed with the CELESTE Cherenkov telescope towards the galaxy M31. We propose a statistical method to reconstruct spectra, mandatory to discriminate classical and exotic spectra. The M31 data analysis enables the extraction of an upper limit on the gamma ray flux, which is the first worldwide for a galaxy in the energy range 50-500 GeV, and whose astrophysical interest goes beyond indirect searches for Dark Matter. (author)

  12. Basic model of fermion dark matter. Indirect detection of supersymmetric dark matter in {gamma} astronomy with the CELESTE telescope; Modele effectif de matiere noire fermionique. Detection indirecte de matiere noire supersymetrique en astronomie {gamma} avec le telescope CELESTE

    Energy Technology Data Exchange (ETDEWEB)

    Lavalle, J

    2004-10-15

    The purpose of this thesis is to discuss both phenomenological and experimental aspects of Dark Matter, related to its indirect detection with gamma-ray astronomy. In the MSSM (Minimal Supersymmetric Standard Model) framework, neutralinos arise as natural candidates to non-baryonic and Cold Dark Matter, whose gravitational effects manifest in the Universe at different scales. As they are Majorana particles, they may in principle annihilate in high density regions, as the centres of galaxies, and produce gamma rays. Nevertheless, the expected fluxes are basically low compared to experimental sensitivities. After estimating gamma fluxes from M31 and Draco galaxies in the MSSM scheme, we first generalize the MSSM couplings by studying an effective Lagrangian. We show that the only constraint of imposing a relic abundance compatible with recent measurements obviously deplete significantly the gamma ray production, but also that predictions in this effective approach are more optimistic for indirect detection than the MSSM. In a second part, we present the indirect searches for Dark Matter performed with the CELESTE Cherenkov telescope towards the galaxy M31. We propose a statistical method to reconstruct spectra, mandatory to discriminate classical and exotic spectra. The M31 data analysis enables the extraction of an upper limit on the gamma ray flux, which is the first worldwide for a galaxy in the energy range 50-500 GeV, and whose astrophysical interest goes beyond indirect searches for Dark Matter. (author)

  13. Indirect MR arthrography of the shoulder in detection of rotator cuff ruptures

    International Nuclear Information System (INIS)

    Yagci, B.; Manisali, M.; Yilmaz, E.; Oezaksoy, D.; Kovanlikaya, I.; Oezkan, M.; Ekin, A.

    2001-01-01

    The aim of this study was to compare the efficacy of indirect MR arthrography images obtained following intravenous contrast injection and conventional MR imaging in the diagnosis of rotator cuff tears. Twenty-four patients with clinically suspected rotator cuff disease were examined. Conventional MR images and post-contrast indirect MR arthrography images were obtained. All images were evaluated in a blinded fashion by two musculoskeletal radiologist. Results were than analyzed depending on surgical output. The correlation coefficient (Spearman rank correlation test) and the kappa values for agreement between surgery and imaging techniques were calculated. The correlation coefficients between indirect MR arthrography and surgery for reader 1 and reader 2 were 0.9137 and 0.9773, respectively. Whereas the agreement between conventional MR imaging and surgery was moderate (κ=0.383-0.571), the agreement between indirect MR arthrography and surgery was excellent (κ=0.873-0.936). We suggest the use of indirect MR arthrography technique when conventional MR images are equivocal in diagnosis of rotator cuff disease. (orig.)

  14. An effective model for fermion dark matter. Indirect detection of supersymmetric dark matter in astronomy with the CELESTE Telescope

    International Nuclear Information System (INIS)

    Lavalle, Julien

    2004-01-01

    The purpose of this thesis is to discuss both phenomenological and experimental aspects of Dark Matter, related to its indirect detection with gamma-ray astronomy. In the MSSM framework, neutralinos arise as natural candidates to non-baryonic and Cold Dark Matter, whose gravitational effects manifest in the Universe at different scales. As they are Majorana particles, they may in principle annihilate in high density regions, as the centres of galaxies, and produce gamma rays. Nevertheless, the expected fluxes are basically low compared to experimental sensitivities. After estimating gamma fluxes from M31 and Draco galaxies in the MSSM scheme, we first generalize the MSSM couplings by studying an effective Lagrangian. We show that the only constraint of imposing a relic abundance compatible with recent measurements obviously deplete significantly the gamma ray production, but also that predictions in this effective approach are more optimistic for indirect detection than the MSSM. In a second part, we present the indirect searches for Dark Matter performed with the CELESTE Cherenkov telescope towards the galaxy M31. We propose a statistical method to reconstruct spectra, mandatory to discriminate classical and exotic spectra. The M31 data analysis enables the extraction of an upper limit on the gamma ray flux, which is the first worldwide for a galaxy in the energy range 50-500 GeV, and whose astrophysical interest goes beyond indirect searches for Dark Matter. (author)

  15. Basic model of fermion dark matter. Indirect detection of supersymmetric dark matter in γ astronomy with the CELESTE telescope

    International Nuclear Information System (INIS)

    Lavalle, J.

    2004-10-01

    The purpose of this thesis is to discuss both phenomenological and experimental aspects of Dark Matter, related to its indirect detection with gamma-ray astronomy. In the MSSM (Minimal Supersymmetric Standard Model) framework, neutralinos arise as natural candidates to non-baryonic and Cold Dark Matter, whose gravitational effects manifest in the Universe at different scales. As they are Majorana particles, they may in principle annihilate in high density regions, as the centres of galaxies, and produce gamma rays. Nevertheless, the expected fluxes are basically low compared to experimental sensitivities. After estimating gamma fluxes from M31 and Draco galaxies in the MSSM scheme, we first generalize the MSSM couplings by studying an effective Lagrangian. We show that the only constraint of imposing a relic abundance compatible with recent measurements obviously deplete significantly the gamma ray production, but also that predictions in this effective approach are more optimistic for indirect detection than the MSSM. In a second part, we present the indirect searches for Dark Matter performed with the CELESTE Cherenkov telescope towards the galaxy M31. We propose a statistical method to reconstruct spectra, mandatory to discriminate classical and exotic spectra. The M31 data analysis enables the extraction of an upper limit on the gamma ray flux, which is the first worldwide for a galaxy in the energy range 50-500 GeV, and whose astrophysical interest goes beyond indirect searches for Dark Matter. (author)

  16. Novel indirect enzyme-linked immunosorbent assay (ELISA) method to detect Total E. coli in water environment

    International Nuclear Information System (INIS)

    Wang Na; He Miao; Shi Hanchang

    2007-01-01

    In order to establish ELISA (enzyme-linked immunosorbent assay) method to detect Total E. coli in water environment, E. coli multi-characters antigens in water environment were prepared according to the characters of kinds of E. coli serotypes, including antigen of whole cell, antigen of disrupted whole cell, somatic antigen, flagellar antigen and fimbrial antigen. Total E. coli polyclonal antibodies were obtained from the New Zealand rabbits immunized with these five antigens, respectively. Antibodies generated in this research are with high titers and good purity, can conjugate with antigens, specifically, stably and strongly. Indirect ELISA shows the titers of antibody of whole cell and antibody of disrupted whole cell are both over 1 x 10 5 . The cross-reactivity of the antibody is from 12 to 30% which indicate the specificity of the antibody against Total E. coli. Based on these antibodies, we established indirect ELISA method to detect Total E. coli in water environment. The matrix effects were studied and the results show that there is no significant influence by all the factors. The ELISA result shows that the detection limitation could be 10 4 CFU (colony forming units) L -1 . The indirect ELISA method developed in this study is well suited for Total E. coli analysis in real water samples as a rapid screen method

  17. Dark matter in dwarf spheroidal galaxies and indirect detection: a review.

    Science.gov (United States)

    Strigari, Louis E

    2018-05-01

    Indirect dark matter searches targeting dwarf spheroidal galaxies (dSphs) have matured rapidly during the past decade. This has been because of the substantial increase in kinematic data sets from the dSphs, the new dSphs that have been discovered, and the operation of the Fermi-LAT and many ground-based gamma-ray experiments. Here we review the analysis methods that have been used to determine the dSph dark matter distributions, in particular the 'J-factors', comparing and contrasting them, and detailing the underlying systematics that still affect the analysis. We discuss prospects for improving measurements of dark matter distributions, and how these interplay with future indirect dark matter searches.

  18. Dark matter in dwarf spheroidal galaxies and indirect detection: a review

    Science.gov (United States)

    Strigari, Louis E.

    2018-05-01

    Indirect dark matter searches targeting dwarf spheroidal galaxies (dSphs) have matured rapidly during the past decade. This has been because of the substantial increase in kinematic data sets from the dSphs, the new dSphs that have been discovered, and the operation of the Fermi-LAT and many ground-based gamma-ray experiments. Here we review the analysis methods that have been used to determine the dSph dark matter distributions, in particular the ‘J-factors’, comparing and contrasting them, and detailing the underlying systematics that still affect the analysis. We discuss prospects for improving measurements of dark matter distributions, and how these interplay with future indirect dark matter searches.

  19. Variability in the recognition of distinctive immunofluorescence patterns in different brands of HEp-2 cell slides

    Directory of Open Access Journals (Sweden)

    Alessandra Dellavance

    2013-06-01

    Full Text Available INTRODUCTION: Indirect immunofluorescence on HEp-2 cells is considered the gold standard for the detection of autoantibodies against cellular antigens. However, the culture conditions, cell fixation and permeabilization processes interfere directly in the preservation and spatial distribution of antigens. Therefore, one can assume that certain peculiarities in the processing of cellular substrate may affect the recognition of indirect immunofluorescence patterns associated with several autoantibodies. OBJECTIVE: To evaluate a panel of serum samples representing nuclear, nucleolar, cytoplasmic, mitotic apparatus, and chromosome plate patterns on HEp-2 cell substrates from different suppliers. MATERIALS AND METHODS: Seven blinded observers, independent from the three selected reference centers, evaluated 17 samples yielding different nuclear, nucleolar, cytoplasmic and mitotic apparatus patterns on HEp-2 cell slides from eight different brands. The slides were coded to maintain confidentiality of both brands and participating centers. RESULTS: The 17 HEp-2 cell patterns were identified on most substrates. Nonetheless, some slides showed deficit in the expression of several patterns: nuclear coarse speckled/U1-ribonucleoprotein associated with antibodies against RNP (U1RNP, centromeric protein F (CENP-F, proliferating cell nuclear antigen (PCNA, cytoplasmic fine speckled associated with anti-Jo-1 antibodies (histidyl synthetase, nuclear mitotic apparatus protein 1 (NuMA-1 and nuclear mitotic apparatus protein 2 (NuMA-2. CONCLUSION: Despite the overall good quality of the assessed HEp-2 substrates, there was considerable inconsistency in results among different commercial substrates. The variations may be due to the evaluated batches, hence generalizations cannot be made as to the respective brands. It is recommended that each new batch or new brand be tested with a panel of reference sera representing the various patterns.

  20. A machine learning approach for indirect human presence detection using IoT devices

    OpenAIRE

    Madeira, Rui Nuno Neves

    2016-01-01

    The recent increased democratization of technology led to the appearance of new devices dedicated to the improvement of our daily living and working spaces, capable of being remotely controlled through the internet and interoperability with other systems. In this context, human presence detection is fundamental for several purposes, such has: further automization, usage pattern learning, problem detection (illness, or intruder), etc. Current intrusion detection devices usual...

  1. Detection of Ganoderic Acid A in Ganoderma lingzhi by an Indirect Competitive Enzyme-Linked Immunosorbent Assay.

    Science.gov (United States)

    Sakamoto, Seiichi; Kohno, Toshitaka; Shimizu, Kuniyoshi; Tanaka, Hiroyuki; Morimoto, Satoshi

    2016-05-01

    Ganoderma is a genus of medicinal mushroom traditionally used for treating various diseases. Ganoderic acid A is one of the major bioactive Ganoderma triterpenoids isolated from Ganoderma species. Herein, we produced a highly specific monoclonal antibody against ganoderic acid A (MAb 12 A) and developed an indirect competitive ELISA for the highly sensitive detection of ganoderic acid A in Ganoderma lingzhi, with a limit of detection of 6.10 ng/mL. Several validation analyses support the accuracy and reliability of the developed indirect competitive ELISA for use in the quality control of Ganoderma based on ganoderic acid A content. Furthermore, quantitative analysis of ganoderic acid A in G. lingzhi revealed that the pileus exhibits the highest ganoderic acid A content compared with the stipe and spore of the fruiting body; the best extraction efficiency was found when 50 % ethanol was used, which suggests the use of a strong liquor to completely harness the potential of Ganoderma triterpenoids in daily life. Georg Thieme Verlag KG Stuttgart · New York.

  2. Guidelines to indirectly measure and enhance detection efficiency of stationary PIT tag interrogation systems in streams

    Science.gov (United States)

    Connolly, Patrick J.; Wolf, Keith; O'Neal, Jennifer S.

    2010-01-01

    With increasing use of passive integrated transponder (PIT) tags and reliance on stationary PIT tag interrogation systems to monitor fish populations, guidelines are offered to inform users how best to use limited funding and human resources to create functional systems that maximize a desired level of detection and precision. The estimators of detection efficiency and their variability as described by Connolly et al. (2008) are explored over a span of likely performance metrics. These estimators were developed to estimate detection efficiency without relying on a known number of fish passing the system. I present graphical displays of the results derived from these estimators to show the potential efficiency and precision to be gained by adding an array or by increasing the number of PIT-tagged fish expected to move past an interrogation system.

  3. Climate and infectious disease: use of remote sensing for detection of Vibrio cholerae by indirect measurement

    Science.gov (United States)

    Lobitz, B.; Beck, L.; Huq, A.; Wood, B.; Fuchs, G.; Faruque, A. S.; Colwell, R.

    2000-01-01

    It has long been known that cholera outbreaks can be initiated when Vibrio cholerae, the bacterium that causes cholera, is present in drinking water in sufficient numbers to constitute an infective dose, if ingested by humans. Outbreaks associated with drinking or bathing in unpurified river or brackish water may directly or indirectly depend on such conditions as water temperature, nutrient concentration, and plankton production that may be favorable for growth and reproduction of the bacterium. Although these environmental parameters have routinely been measured by using water samples collected aboard research ships, the available data sets are sparse and infrequent. Furthermore, shipboard data acquisition is both expensive and time-consuming. Interpolation to regional scales can also be problematic. Although the bacterium, V. cholerae, cannot be sensed directly, remotely sensed data can be used to infer its presence. In the study reported here, satellite data were used to monitor the timing and spread of cholera. Public domain remote sensing data for the Bay of Bengal were compared directly with cholera case data collected in Bangladesh from 1992-1995. The remote sensing data included sea surface temperature and sea surface height. It was discovered that sea surface temperature shows an annual cycle similar to the cholera case data. Sea surface height may be an indicator of incursion of plankton-laden water inland, e.g., tidal rivers, because it was also found to be correlated with cholera outbreaks. The extensive studies accomplished during the past 25 years, confirming the hypothesis that V. cholerae is autochthonous to the aquatic environment and is a commensal of zooplankton, i.e., copepods, when combined with the findings of the satellite data analyses, provide strong evidence that cholera epidemics are climate-linked.

  4. Use of Indirect Calorimetry to Detect Overfeeding in Critically Ill Children: Finding the Appropriate Definition.

    Science.gov (United States)

    Kerklaan, Dorian; Hulst, Jessie M; Verhoeven, Jennifer J; Verbruggen, Sascha C A T; Joosten, Koen F M

    2016-10-01

    Overfeeding during critical illness is associated with adverse effects such as metabolic disturbances and increased risk of infection. Because of the lack of sound studies with clinical endpoints, overfeeding is arbitrarily defined as the ratio caloric intake/measured resting energy expenditure (mREE) or alternatively as a comparison of measured respiratory quotient (RQ) to the predicted RQ based on the macronutrient intake (RQmacr). We aimed to compare definitions of overfeeding in critically ill mechanically ventilated children based on mREE, RQ, and caloric intake to find an appropriate definition. Indirect calorimetry measurements were performed in 78 mechanically ventilated children, median age 6.3 months. Enteral and/or parenteral nutrition was provided according to the local guidelines. Definitions used to indicate overfeeding were the ratio caloric intake/mREE of >110% and >120% and by the measured RQ > RQmacr + 0.05. The proportion of patients identified as overfed varied widely depending on the definition used, ranging from 22% (RQ > RQmacr + 0.05), to 40% and 50% (caloric intake/mREE of >120% and >110%, respectively). Linear regression analysis showed that all patients would be identified as overfed with the definition RQ > RQmacr + 0.05 when the ratio caloric intake/mREE exceeded 165%. Caloric intake was higher in children with a standard deviation-score weight for age definition applied. These currently used definitions fail to take into account several relevant factors affecting metabolism during critical illness and are therefore not generally applicable to the pediatric intensive care unit population.

  5. Application of cholesterol determination method to indirectly detect meat and bone meals in ruminant feeds

    Directory of Open Access Journals (Sweden)

    Cecília M. Bandeira

    2013-01-01

    Full Text Available The aim of this study was to verify the presence of meat and bone meal (MBM in ruminant feed, by identifying the cholesterol using gas chromatography with a flame ionization detector. The proposed method demonstrated precision, trueness, and capability to detect MBM in the ruminant feed.

  6. Detection of understory bamboo in giant panda habitats using an indirect remote sensing approach

    NARCIS (Netherlands)

    Bian, B.M.; Wang, T.; Liu, Y.F.; Fei, T.; Skidmore, A.K.

    2007-01-01

    The bamboo is the exclusive food of the wild giant pandas. Detection of the bamboo forest in giant panda habitat will help scientists further understand the spatial distribution pattern of giant pandas and their habitats. Moreover, it provides crucial scientific evidence for estimating habitat

  7. Minimizing the t1-noise when using an indirect 1H high-resolution detection of unlabeled samples.

    Science.gov (United States)

    Shen, M; Wegner, S; Trébosc, J; Hu, B; Lafon, O; Amoureux, J P

    2017-10-01

    The most utilized through-space correlation 1 H-{X} methods with proton indirect detection use two consecutive transfers, 1 H → X and then X →  1 H, with the evolution time t 1 in the middle. When the X isotope is not 100% naturally abundant (NA), only the signal of the protons close to these isotopes is modulated by the 1 H-X dipolar interactions. This signal is theoretically disentangled with phase-cycling from the un-modulated one. However, this separation is never perfect and it may lead to t 1 -noise in case of isotopes with very small NA, such as 13 C or even worse 15 N. One way to reduce this t 1 -noise is to minimize, 'purge', during t 1 the un-modulated 1 H magnetization before trying to suppress it with phase-cycling. We analyze experimentally several sequences following the HORROR condition, which allow purging the 1 H transverse magnetization. The comparison is made at three spinning speeds, including very fast ones for 1 H resolution: 27.75, 55.5 and 111 kHz. We show (i) that the efficiency of this purging process increases with the spinning speed, and (ii) that the best recoupling sequences are the two simplest ones: XY and S 1  = SR2 1 2 . We then compare the S/N that can be achieved with the two most used 1 H-{X} 2D methods, called D-HMQC and CP-CP. The only difference in between these two methods is that the transfers are done with either two π/2-pulses on X channel (D-HMQC), or two Cross-Polarization (CP) transfers (CP-CP). The first method, D-HMQC, is very robust and should be preferred when indirectly detecting nuclei with high NA. The second method, CP-CP, (i) requires experimental precautions to limit the t 1 -noise, and (ii) is difficult to use with quadrupolar nuclei because the two CP transfers are then not efficient nor robust. However, CP-CP is presently the best method to indirectly detect isotopes with small NA, such as 13 C and 15 N. Copyright © 2017 Elsevier Inc. All rights reserved.

  8. Biostatistical analysis of quantitative immunofluorescence microscopy images.

    Science.gov (United States)

    Giles, C; Albrecht, M A; Lam, V; Takechi, R; Mamo, J C

    2016-12-01

    Semiquantitative immunofluorescence microscopy has become a key methodology in biomedical research. Typical statistical workflows are considered in the context of avoiding pseudo-replication and marginalising experimental error. However, immunofluorescence microscopy naturally generates hierarchically structured data that can be leveraged to improve statistical power and enrich biological interpretation. Herein, we describe a robust distribution fitting procedure and compare several statistical tests, outlining their potential advantages/disadvantages in the context of biological interpretation. Further, we describe tractable procedures for power analysis that incorporates the underlying distribution, sample size and number of images captured per sample. The procedures outlined have significant potential for increasing understanding of biological processes and decreasing both ethical and financial burden through experimental optimization. © 2016 The Authors Journal of Microscopy © 2016 Royal Microscopical Society.

  9. Indirect glyphosate detection based on ninhydrin reaction and surface-enhanced Raman scattering spectroscopy

    Science.gov (United States)

    Xu, Meng-Lei; Gao, Yu; Li, Yali; Li, Xueliang; Zhang, Huanjie; Han, Xiao Xia; Zhao, Bing; Su, Liang

    2018-05-01

    Glyphosate is one of the most commonly-used and non-selective herbicides in agriculture, which may directly pollute the environment and threaten human health. A simple and effective approach to assessment of its damage to the natural environment is thus quite necessary. However, traditional chromatography-based detection methods usually suffer from complex pretreatment procedures. Herein, we propose a simple and sensitive method for the determination of glyphosate by combining ninhydrin reaction and surface-enhanced Raman scattering (SERS) spectroscopy. The product (purple color dye, PD) of the ninhydrin reaction is found to SERS-active and directly correlate with the glyphosate concentration. The limit of detection of the proposed method for glyphosate is as low as 1.43 × 10- 8 mol·L- 1 with a relatively wider linear concentration range (1.0 × 10- 7-1.0 × 10- 4 mol·L- 1), which demonstrates its great potential in rapid, highly sensitive concentration determination of glyphosate in practical applications for safety assessment of food and environment.

  10. The indirect detection of bleach (sodium hypochlorite) in beverages as evidence of product tampering.

    Science.gov (United States)

    Jackson, David S; Crockett, David F; Wolnik, Karen A

    2006-07-01

    Bleach (sodium hypochlorite) has been identified as the adulterant in a relatively large number of product tamperings that have been investigated by the Forensic Chemistry Center (FCC) of the U.S. Food and Drug Administration. In this work, household bleach was added to 23 different beverages at each of three levels. The impact of sodium hypochlorite on these beverages over a 13-day study period was evaluated using the following techniques: diphenylamine spot test for oxidizing agents, potassium iodide-starch test paper for oxidizing agents, pH, iodometric titration for quantitating hypochlorite, ion chromatography for chloride and chlorate quantitation, automated headspace sampling with gas chromatography-flame ionization detection (GC-FID) for determination of chloroform, and visual and organoleptic observations. This study has shown that hypochlorite is fragile when added to most common beverages and typically breaks down either partially or completely over time. In cases where a beverage is suspected of being adulterated with bleach but tests for hypochlorite are negative, it is still possible to characterize the product to demonstrate that the results are consistent with the addition of bleach. An adulterated product will give a positive test for oxidizing agents using the diphenylamine spot test. It is likely that the pH of the adulterated product will be higher than a control of that product. Ion chromatographic analysis shows elevated chloride and chlorate as compared with a control. And, chloroform may also be detected by GC-FID especially if the beverage that was adulterated contains citric acid.

  11. Cosmic Ray Experiments and the Implications for Indirect Detection of Dark Matter

    Science.gov (United States)

    Mitchell, John W.; Ormes, Jonathan F.; Streitmatter, Robert E.

    2013-01-01

    Detection of cosmic-ray antiprotons was first reported by Golden et al. in 1979 and their existence was firmly established by the BESS and IMAX collaborations in the early 1990s. Increasingly precise measurements of the antiproton spectrum, most recently from BESS-Polar and PAMELA, have made it an important tool for investigating cosmic-ray transport in the galaxy and heliosphere and for constraining dark-matter models. The history of antiproton measurements will be briefly reviewed. The current status will be discussed, focusing on the results of BESS-Polar II and their implications for the possibility of antiprotons from primordial black hole evaporation. The current results of the BESS-Polar II antihelium search are also presented.

  12. Development and validation of an indirect pulsed electrochemical detection method for monitoring the inhibition of Abl1 tyrosine kinase.

    Science.gov (United States)

    Chen, Hui; Wang, Xu; Chopra, Shruti; Adams, Erwin; Van Schepdael, Ann

    2014-03-01

    A new method for monitoring the enzyme inhibition of Abl1 tyrosine kinase by liquid chromatography-indirect pulsed electrochemical detection (LC-InPED) was developed. In this method, adsorption of a peptide analyte at the noble metal electrode suppresses the oxidation of polyols under alkaline condition to elicit an indirect response resulting in a negative peak of the target peptide. Among the reagents tested, D-gluconic acid sodium salt gave the best overall signal to noise (S/N) values for the indirect detection of p-Abltide, the product of Abl1 enzymatic reaction. 50 μM D-gluconic acid sodium salt dissolved in a mixture of 78% water-22% acetonitrile-0.03% trifluoroacetic acid (TFA) was used as the mobile phase. Chromatographic separation was achieved on an Alltima C18 (I.D. 5 μm; 250 mm × 4.6 mm) column with the mobile phase flow rate of 0.5 ml/min. 0.5M sodium hydroxide was added post-column to maintain alkaline conditions in the PED cell. The limit of quantification (LOQ) was 0.2 μM for p-Abltide, which was about 50-fold lower than direct PED analysis. The residual plot of the linear calibration curve indicated a good fit with a linear model within the investigated concentration range of p-Abltide. Intra- and inter-day precision was not more than 6.5% and accuracy was from -5.75% to +1.54%. The validated LC-InPED method was successfully applied for monitoring of p-Abltide in Abl1 enzyme reaction and the inhibition study of Abl1. The determined IC50 values of model inhibitors, imatinib, nilotinib and dasatinib, were 601.4 nM (R(2)=0.99), 32.3 nM (R(2)=0.99) and 1.3 nM (R(2)=0.98), respectively. These results were consistent with literature data. To the best of our knowledge this is the first time a LC-InPED method has been used to monitor an enzyme reaction. Copyright © 2013 Elsevier B.V. All rights reserved.

  13. Capillary electrophoresis with indirect UV detection for the determination of stabilizers and citrates present in human albumin solutions.

    Science.gov (United States)

    Jaworska, Małgorzata; Cygan, Paulina; Wilk, Małgorzata; Anuszewska, Elzbieta

    2009-08-15

    Sodium caprylate and N-acetyltryptophan are the most frequently used stabilizers that protect the albumin from aggregation or heat induced denaturation. In turn citrates - excipients remaining after fractionation process - can be treated as by-product favoring leaching aluminum out of glass containers whilst albumin solution is stored. With ionic nature these substances have all the markings of a subject for capillary electrophoresis analysis. Thus CE methods were proposed as new approach for quality control of human albumin solution in terms of determination of stabilizers and citrates residue. Human albumin solutions both 5% and 20% from various manufacturers were tested. Indirect detection mode was set to provide sufficient detectability of analytes lacking of chromophores. As being anions analytes were separated with reversed electroosmotic flow. As a result of method optimization two background electrolytes based on p-hydroxybenzoic acid and 2,6-pyridinedicarboxylic acid were selected for stabilizers and citrates separation, respectively. The optimized methods were successfully validated. For citrates that require quantification below 100microM the method demonstrated the precision less than 4% and the limit of detection at 4microM. In order to check the new methods accuracy and applicability the samples were additionally tested with selected reference methods. The proposed methods allow reliable quantification of stabilizers and citrates in human albumin solution that was confirmed by method validation as well as result comparison with reference methods. The CE methods are considered to be suitable for quality control yet simplifying and reducing cost of analysis.

  14. Indirect fluorescence detection of native amino acids in capillary zone electrophoresis

    Energy Technology Data Exchange (ETDEWEB)

    Kuhr, W.G.; Yeung, E.S.

    1988-09-01

    Amino acids are but one of several important classes of small chemical compounds in biological chemistry that have an inherent lack of analytically useful physical properties. Amino acids, peptides, fatty acids, sugars, many mono-, di-, and tricarboxylic acids, and phosphorylated intermediates in glycolysis and metabolism show little, if any, UV or visible absorption, fluorescence, or electrochemical activity. As the emphasis of biochemical research shifts to smaller samples where, for example, picomolar quantities of amino acids are analyzed in gas phase protein sequencing or in microliter samples of the extracellular fluid of the mammalian brain, the analytical problem becomes even more challenging due to the small volume of sample available for analysis. In this work, laser-induced fluorescence spectroscopy is performed on-column to detect the bands separated with capillary zone electrophoresis (CZE). CZE is an instrumental form of zone electrophoresis where chemical species are separated purely on the basis of their electrophoretic mobility, since no supporting gel is utilized. Both anions and cations can be separated in the same run because of the large electroosmotic flow generated in small diameter capillaries. This technique has already been used successfully in the rapid, efficient separation of dansyl-amino acids.

  15. Development and validation of an indirect Enzyme-linked Immunosorbent Assay for the detection of antibodies against Schmallenberg virus in blood samples from ruminants

    NARCIS (Netherlands)

    Heijden, van der H.M.J.F.; Bouwstra, R.J.; Mars, M.H.; Poel, van der W.H.M.; Wellenberg, G.J.; Maanen, van C.

    2013-01-01

    To detect Schmallenberg virus (SBV) infections in ruminants and to perform SBV epidemiological studies a cost-effective serological test is required. For these purposes an indirect whole virus Enzyme-linked Immunosorbent Assay (ELISA) for detection of SBV specific antibodies in ruminant blood

  16. DMBLC: An Indirect Urban Impervious Surface Area Extraction Approach by Detecting and Masking Background Land Cover on Google Earth Image

    Directory of Open Access Journals (Sweden)

    Min Huang

    2018-05-01

    Full Text Available Implying the prosperity and development of the city, impervious surface area (ISA is playing an increasingly important role in ecological processes, microclimate, material and energy flows, and urban flood. The free sub-meter resolution Google Earth image, which is integrated by several high spatial resolution data, appears to have potential for high-resolution ISA extraction, where present study is rare and performances remain to be improved. Due to the high spatial and spectral variation of the urban environment as well as confusion between ISA and soil, the accurate delineating of ISA with traditional (direct methods can be costly and time-consuming, which is in a word resource-intensive. However, this paper presents a novel indirect ISA extraction conceptual model and a new detecting and masking background land cover (DMBLC approach that: uses a freely available, high-resolution dataset; requires a reduced set of training samples; and consists of relatively simple, common, and feasible image processing steps. The key characteristic of DMBLC is to detect the background of ISA (vegetation, soil, and water accurately and obtain the ISA by masking the background. The approach relies on background detection to avoid the predicaments of direct ISA extraction. Water can be directly gained by water body vector data, in DMBLC; mixture tuned matched filtering (MTMF is exploited to detect vegetation and soil, image segmentation is used to mitigate the spectral variation problem within the same land cover, and segment rectangularity reduces the confusion between ISA and soil. From experiments in a core area of Fuzhou, China, the DMBLC approach reached high performance and outperformed the powerful traditional support vector machines (SVM method (overall accuracy of 94.45% and Kappa coefficient of 0.8885, compared to 86.44% and 0.7329, respectively. From the comparison of different levels of complexity within the inner processing steps, it is confirmed

  17. Deteccion de Chlamydia trachomatis en muestras uretrales mediante inmunofluorescencia directa Detecção de Chlamydia trachomatis em amostras uretrais mediante imunofluorescência direta Detection of Chlamydia trachomatis in urethral samples by means of direct immunofluorescence

    Directory of Open Access Journals (Sweden)

    Myra Wilson Schuster

    1989-12-01

    Full Text Available Se estudiaron 82 pacientes con uretritis para la búsqueda de Chlamydia trachomatis mediante inmunofluorescencia directa, Neisscria gonorrhoeae, Mycoplastna y Ureaplasma mediante métodos estándar. Se encontró un 19,5% de Chlamydia trachomatis y en 11 de ellos (68,8% se encontró asociada a otras bacterias y estos pacientes presentó una secreción escasa-gelatinosa.Em 82 doentes com uretrite foi pesquisada a presença de Chlamydia trachomatis, utilizando a prova da imunofluorescência direta, e de Neisseria gonorrhoeae, Mycoplasma e Ureaplasma, utilizando os métodos padrões. Ch. trachomatis foi encontrada em 19,5% dos casos, sendo que em 11 deles (68,8% observou-se associação entre Chlamydia e as outras bactérias pesquisadas. Nesses pacientes observou-se presença de secreção uretral escassa e de aspecto gelatinoso.The presence of Chlamydia trachomatis was studied by the direct immunofluorescence test, as also was that of Neisseria gonorrhoeae, Mycoplasma and Ureaplasma by the standard methods, in 82 patients with urethral discharge. Ch. trachomatis was found in 19.5% (16 of the cases and in 11 of them (68.8% there was association with the other bacteria investigated. This eleven patients presented a scanty gelatinous discharge.

  18. Diagnostic significance of colloid body deposition in direct immunofluorescence

    Directory of Open Access Journals (Sweden)

    Chularojanamontri Leena

    2010-01-01

    Full Text Available Background: Colloid bodies (CB in direct immunofluorescence (DIF studies are usually found in interface dermatitis. Furthermore, CB can be found in various skin diseases and even in normal skin. Aim: To evaluate the diagnostic value of CB deposits in DIF studies. Methods: From 1996-2007, data from 502 patients where DIF studies showed immunoreactants at CB were enrolled. The definite diagnoses of these patients were based on clinical, histopathological and immunofluorescent findings. The results of DIF studies were analyzed. Results: Immunoreactants at CB were detected in 44.4%, 43.8%, 4.2%, 3.8%, and 2.2% of interface dermatitis, vasculitis, autoimmune vesiculobullous disease, panniculitis, and scleroderma/morphea, respectively. The most common immunoreactant deposit of all diseases was Immunoglobulin M (IgM. Brighter intensity and higher quantity of CB was detected frequently in the group with interface dermatitis. Conclusions: Immunoreactant deposits at CB alone can be found in various diseases but a strong intensity and high quantity favor the diagnosis of interface dermatitis. CB plus dermoepidermal junction (DEJ deposits are more common in interface dermatitis than any other disease. Between lichen planus (LP and discoid lupus erythematosus (DLE, CB alone is more common in LP; whereas, CB plus DEJ and superficial blood vessel (SBV is more common in DLE. The most common pattern in both diseases is CB plus DEJ. The quantity and intensity of CB in LP is higher than in DLE.

  19. [Laboratory diagnosis of genital herpes--direct immunofluorescence method].

    Science.gov (United States)

    Majewska, Anna; Romejko-Wolniewicz, Ewa; Zareba-Szczudlik, Julia; Kilijańczyk, Marek; Gajewska, Małgorzata; Młynarczyk, Grazyna

    2013-07-01

    Aim of the study was to determine clinical usefulness of direct immunofluorescence method in the laboratory diagnosis of genital herpes in women. Overall 187 anogenital swabs were collected from 120 women. Using a dacron-tipped applicator 83 swabs were collected from women suspected of genital herpes and 104 from patients with no signs of genital infection. All samples were tested using cell culture (Vero cell line) and then direct immunofluorescence method (DIF) for the identification of antigens of herpes simplex viruses: HSV-1 and HSV-2. Characteristic cytopathic effect (CPE), indicative of alphaherpesvirus infection, was observed in 43.4% of cultures with clinical specimens collected from women with suspected genital herpes and in 29.8% of cultures of clinical specimens taken from patients with no clinical symptoms of genital herpes. Herpes simplex viruses were determined in 73 samples by direct immunofluorescence method after amplification of the virus in cell culture. The DIF test confirmed the diagnosis based on the microscopic CPE observation in 85%. In 15% of samples (taken from pregnant women without clinical signs of infection) we reported positive immunofluorescence in the absence of CPE. The frequency of antigen detection was statistically significantly higher in samples that were positive by culture study (chi-square test with Yates's correction, p genital herpes in swabs taken from the vestibule of the vagina and the vulva. However, there was no statistically significant difference in the frequency of detection of Herpes Simplex Virus antigens in specimens from different parts of the genital tract in both groups of women (chi-square test, p > 0.05). In our study HHV-1 was the main causative agent of genital herpes. The growing worldwide prevalence of genital herpes, challenges with the clinical diagnosis, and availability of effective antiviral therapy are the main reasons for a growing interest in rapid, proper laboratory diagnosis of infected

  20. Correlation of antinuclear antibody immunofluorescence patterns with immune profile using line immunoassay in the Indian scenario

    Directory of Open Access Journals (Sweden)

    Sebastian Wendy

    2010-07-01

    Full Text Available Background: Immunity status, individual response to disease and types of antibodies produced are well known to vary from person to person, place to place and probably from population to population. A broad spectrum of specific auto antibodies that have so far been associated with specific rheumatic diseases, as noted in Western literature, has been well taken as a reference standard all over the world. There is neither research work nor any data correlating the auto antibodies and their antinuclear antibody (ANA patterns with the immunoprofile in the Indian population to date. Aims: To understand a definite association between ANA patterns and specific antibodies in the serum in the Indian study population and to document similarities / differences with the West. Settings and Design: This prospective and retrospective double blind study was undertaken on the South Indian population referred for ANA testing by Indirect Immunofluorescence method and by immunoline methods. Materials and Methods: Serum samples of patients from a random South Indian population who sought medical help for rheumatic disease were subjected for ANA testing by indirect immunofluorescence (IIF method and line immunoassay during the study period of 27 months. Serum samples were processed in dilution of 1:100 using HEp - 2010 / liver biochip (Monkey (EUROIMMUN AG. The serum samples which were further processed for line immunoassay were treated in 1:100 dilution on nylon strips coated with recombinant and purified antigens as discrete lines with plastic backing (EUROIMMUN AG coated with antigens nRNP / Sm, Sm, SSA, Ro-52, SSB, Scl-70, PM-Scl, PCNA, Jo-1, CENP-B, dsDNA, nucleosomes, histones, ribosomal protein-P, anti-mitochondrial antibodies (AMA-M2 along with a control band. The analysis was done by comparing the intensity of the reaction with positive control line by image analysis. Results: The antinuclear antibody indirect immunofluorescence (ANA - IIF patterns obtained

  1. Indirect ultraviolet detection of alkaline earth metal ions using an imidazolium ionic liquid as an ultraviolet absorption reagent in ion chromatography.

    Science.gov (United States)

    Liu, Yong-Qiang; Yu, Hong

    2017-04-01

    A convenient and versatile method was developed for the separation and detection of alkaline earth metal ions by ion chromatography with indirect UV detection. The chromatographic separation of Mg 2+ , Ca 2+ , and Sr 2+ was performed on a carboxylic acid base cation exchange column using imidazolium ionic liquid/acid as the mobile phase, in which the imidazolium ionic liquid acted as an UV-absorption reagent. The effects of imidazolium ionic liquids, detection wavelength, acids in the mobile phase, and column temperature on the retention of Mg 2+ , Ca 2+ , and Sr 2+ were investigated. The main factors influencing the separation and detection were the background UV absorption reagent and the concentration of hydrogen ion in ion chromatography with indirect UV detection. The successful separation and detection of Mg 2+ , Ca 2+ , and Sr 2+ within 14 min were achieved using the selected chromatographic conditions, and the detection limits (S/N = 3) were 0.06, 0.12, and 0.23 mg/L, respectively. A new separation and detection method of alkaline earth metal ions by ion chromatography with indirect UV detection was developed, and the application range of ionic liquids was expanded. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  2. Ion chromatography with the indirect ultraviolet detection of alkali metal ions and ammonium using imidazolium ionic liquid as ultraviolet absorption reagent and eluent.

    Science.gov (United States)

    Liu, Yong-Qiang; Yu, Hong

    2016-08-01

    Indirect ultraviolet detection was conducted in ultraviolet-absorption-agent-added mobile phase to complete the detection of the absence of ultraviolet absorption functional group in analytes. Compared with precolumn derivatization or postcolumn derivatization, this method can be widely used, has the advantages of simple operation and good linear relationship. Chromatographic separation of Li(+) , Na(+) , K(+) , and NH4 (+) was performed on a carboxylic acid base cation exchange column using imidazolium ionic liquid/acid/organic solvent as the mobile phase, in which imidazolium ionic liquids acted as ultraviolet absorption reagent and eluting agent. The retention behaviors of four kinds of cations are discussed, and the mechanism of separation and detection are described. The main factors influencing the separation and detection were the background ultraviolet absorption reagent and the concentration of hydrogen ion in the ion chromatography-indirect ultraviolet detection. The successful separation and detection of Li(+) , Na(+) , K(+) , and NH4 (+) within 13 min was achieved using the selected chromatographic conditions, and the detection limits (S/N = 3) were 0.02, 0.11, 0.30, and 0.06 mg/L, respectively. A new separation and analysis method of alkali metal ions and ammonium by ion chromatography with indirect ultraviolet detection method was developed, and the application range of ionic liquid was expanded. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  3. Indirect competitive immunoassay for the detection of fungicide Thiabendazole in whole orange samples by Surface Plasmon Resonance.

    Science.gov (United States)

    Estevez, M-Carmen; Belenguer, Jose; Gomez-Montes, Silvia; Miralles, Javier; Escuela, Alfonso M; Montoya, Angel; Lechuga, Laura M

    2012-12-07

    A highly sensitive and specific SPR-based competitive immunoassay for the detection of Thiabendazole (TBZ) has been developed. An indirect format where a TBZ-protein conjugate is immobilized onto gold surfaces has been selected. Under the optimal conditions, a LOD of 0.67 nM (0.13 μg L(-1)) and an IC(50) of 3.2 nM (0.64 μg L(-1)) have been achieved which are comparable to the values obtained by conventional ELISA. Analysis of real samples has been attempted by first evaluating the influence of complex matrix samples coming from whole oranges and secondly measuring samples containing TBZ previously evaluated by chromatographic methods. A methanolic extraction procedure followed by a simple dilution in assay buffer has proven to be sufficient to measure orange samples using the developed immunoassay with an excellent recovery percentage. The sensitivity and the feasibility of measuring whole orange samples demonstrate the effectiveness and robustness of the SPR biosensor, which can be useful for the determination of TBZ in food at concentrations below the Maximum Residue Levels (MRLs) established by the European legislation.

  4. Development of an Indirect ELISA Based on a Recombinant Chimeric Protein for the Detection of Antibodies against Bovine Babesiosis.

    Science.gov (United States)

    Jaramillo Ortiz, José Manuel; Montenegro, Valeria Noely; de la Fournière, Sofía Ana María; Sarmiento, Néstor Fabián; Farber, Marisa Diana; Wilkowsky, Silvina Elizabeth

    2018-01-23

    The current method for Babesia spp. serodiagnosis based on a crude merozoite antigen is a complex and time-consuming procedure. An indirect enzyme-linked immunosorbent assay (iELISA) based on a recombinant multi-antigen of Babesia bovis (rMABbO) was developed for detection of antibodies in bovines suspected of infection with this parasite. The multi-antigen comprises gene fragments of three previously characterized B. bovis antigens: MSA-2c, RAP-1 and the Heat Shock protein 20 that are well-conserved among geographically distant strains. The cutoff value for the new rMABbo-iELISA was determined using 75 known-positive and 300 known-negative bovine sera previously tested for antibodies to B. bovis by the gold-standard ELISA which uses a merozoite lysate. A cutoff value of ≥35% was determined in these samples by receiver operator characteristic (ROC) curve analysis, showing a sensitivity of 95.9% and a specificity of 94.3%. The rMABbo-iELISA was further tested in a blind trial using an additional set of 263 field bovine sera from enzootic and tick-free regions of Argentina. Results showed a good agreement with the gold standard test with a Cohen's kappa value of 0.76. Finally, the prevalence of bovine babesiosis in different tick enzootic regions of Argentina was analyzed where seropositivity values among 68-80% were obtained. A certain level of cross reaction was observed when samples from B. bigemina infected cattle were analyzed with the new test, which can be attributed to shared epitopes between 2 of the 3 antigens. This new rMABbo-iELISA could be considered a simpler alternative to detect anti Babesia spp. antibodies and appears to be well suited to perform epidemiological surveys at the herd level in regions where ticks are present.

  5. Highly specific and rapid immuno-fluorescent visualization and detection of E. coli O104:H4 with protein-A coated magnetic beads based LST-MUG assay.

    Science.gov (United States)

    Barizuddin, Syed; Balakrishnan, Baskar; Stringer, R Cody; Dweik, Majed

    2015-08-01

    A method combining immunomagnetic separation and fluorescent sensing was developed to detect Escherichia coli (E. coli) O104:H4. The antibody specific to E. coli O104:H4 was immobilized on protein A-coated magnetic beads. This protein-A-anti E. coli O104:H4 complex was used to bind Fluorescein IsoThioCyanate (FITC) labeled E. coli O104:H4 antigen (whole cell) on it. The goal was to achieve a fluorescently detectable protein-A-anti E. coli O104:H4-E. coli O104:H4 complex on the magnetic beads. Fluorescent microscopy was used to image the magnetic beads. The resulting fluorescence on the beads was due to the FITC labeled antigen binding on the protein-A-anti E. coli O104:H4 immobilized magnetic beads. This visually proves the antigen-antibody binding. The fluorescent imaging results were obtained in 2 h if the minimum available bacteria in the sample were at least 10(5) CFU/ml. If no fluorescence was observed on the magnetic beads during fluorescent imaging, it indicates the bacterial concentration in the sample to be too low for it to have bound to the magnetic beads and hence no detection was possible. To detect bacterial concentration less than 10(5) CFU/ml in the sample, an additional step was required for detection. The magnetic bead complex was added to the LST-MUG (lauryl sulfate tryptose-4-methylumbelliferyl-β-D-glucuronide), a signaling reporter. The E. coli O104:H4 grows in LST-MUG and releases β-glucuronidase enzyme. This enzyme cleaves the MUG substrate that produces 4-methylumbelliferone, a highly fluorescent species. This fluorescence was detected using a spectrofluorometer. The emission peak in the fluorescent spectrum was found to be at 450 nm. The lower and upper detection range for this LST-MUG assay was found to be 2.05×10(5)-4.09×10(8) CFU/ml. The results for the LST-MUG assay for concentrations below 10(5) CFU/ml were ascertained in 8h. The advantages of this technique include the specific detection of bacteria without an enrichment step and

  6. Detection of Candida albicans Sap2 in cancer patient serum samples by an indirect competitive enzyme-linked immunosorbent assay for the diagnosis of candidiasis

    Directory of Open Access Journals (Sweden)

    Yicun Wang

    2013-01-01

    Full Text Available Background: The secreted aspartyl proteinases 2 (Sap2 of Candida albicans (C. albicans is a potential marker of candididasis. It is a virulence factor associated with adherence and tissue invasion. Aim: In order to detect Sap2 in clinical sera, we developed an indirect competitive enzyme-linked immunosorbent assay (ELISA. Materials and Methods: Polyclonal antibodies were produced for Sap2 by injecting Sap2 into a New Zealand White inbred rabbit. They could be used at a dilution exceeding 1:1200 in an indirect ELISA, and detected Sap2 concentration up to 1 ng/mL. Results: Of the 286 cancer serum samples tested, 16.8% were found as candidiasis. The test was simple and economical to perform and had a level of sensitivity for detection of low-titer positive sera; thus, it may be proven to be of value in epidemiological studies on candidiasis.

  7. Detection of Candida albicans Sap2 in cancer patient serum samples by an indirect competitive enzyme-linked immunosorbent assay for the diagnosis of candidiasis.

    Science.gov (United States)

    Wang, Yicun; Gao, Xiang; Zhi Gang, J U; Liu, Jingyuan; Dong, Shuai; Wang, Li

    2013-01-01

    The secreted aspartyl proteinases 2 (Sap2) of Candida albicans (C. albicans) is a potential marker of candididasis. It is a virulence factor associated with adherence and tissue invasion. In order to detect Sap2 in clinical sera, we developed an indirect competitive enzyme-linked immunosorbent assay (ELISA). Polyclonal antibodies were produced for Sap2 by injecting Sap2 into a New Zealand White inbred rabbit. They could be used at a dilution exceeding 1:1200 in an indirect ELISA, and detected Sap2 concentration up to 1 ng/mL. Of the 286 cancer serum samples tested, 16.8% were found as candidiasis. The test was simple and economical to perform and had a level of sensitivity for detection of low-titer positive sera; thus, it may be proven to be of value in epidemiological studies on candidiasis.

  8. Use of commercially available rabbit monoclonal antibodies for immunofluorescence double staining

    DEFF Research Database (Denmark)

    Bzorek, M.; Stamp, I.M.; Frederiksen, L.

    2008-01-01

    Immunohistochemistry, that is, the use of polyclonal and monoclonal antibodies to detect cell and tissue antigens at a microscopical level is a powerful tool for both research and diagnostic purposes. Especially in the field of hematologic disease, there is often a need to detect several antigens...... synchronously, and we report here a fast and easy technique for demonstrating more than 1 antigen in 1 slide using immunofluorescence. We have used commercially available rabbit monoclonal antibodies (Cyclin D1, CD3, CD5, CD23, etc.) paired with mouse monoclonal antibodies (CD7, CD20, CD79a, Pax-5, etc.......) for double immunofluorescence labeling on paraffin-embedded tissue sections. Commercially available rabbit monoclonal antibodies in combination with mouse monoclonal antibodies proved useful in double immunofluorescence labeling on paraffin-embedded tissue, and all combinations used yielded excellent results...

  9. Analysis of the imaging performance in indirect digital mammography detectors by linear systems and signal detection models

    International Nuclear Information System (INIS)

    Liaparinos, P.; Kalyvas, N.; Kandarakis, I.; Cavouras, D.

    2013-01-01

    Purpose: The purpose of this study was to provide an analysis of imaging performance in digital mammography, using indirect detector instrumentation, by combining the Linear Cascaded Systems (LCS) theory and the Signal Detection Theory (SDT). Observer performance was assessed, by examining frequently employed detectors, consisting of phosphor-based X-ray converters (granular Gd 2 O 2 S:Tb and structural CsI:Tl), coupled with the recently introduced complementary metal-oxide-semiconductor (CMOS) sensor. By applying combinations of various irradiation conditions (filter-target and exposure levels at 28 kV) on imaging detectors, our study aimed to find the optimum system set-up for digital mammography. For this purpose, the signal to noise transfer properties of the medical imaging detectors were examined for breast carcinoma detectability. Methods: An analytical model was applied to calculate X-ray interactions within software breast phantoms and detective media. Modeling involved: (a) three X-ray spectra used in digital mammography: 28 kV Mo/Mo (Mo: 0.030 mm), 28 kV Rh/Rh (Rh: 0.025 mm) and 28 kV W/Rh (Rh: 0.060 mm) at different entrance surface air kerma (ESAK) of 3 mGy and 5 mGy, (b) a 5 cm thick Perspex software phantom incorporating a small Ca lesion of varying size (0.1–1 cm), and (c) two 200 μm thick phosphor-based X-ray converters (Gd2O2S:Tb, CsI:Tl), coupled to a CMOS based detector of 22.5 μm pixel size. Results: Best (lowest) contrast threshold (CT) values were obtained with the combination: (i) W/Rh target-filter, (ii) 5 mGy (ESAK), and (iii) CsI:Tl-CMOS detector. For lesion diameter 0.5 cm the CT was found improved, in comparison to other anode/filter combinations, approximately 42% than Rh/Rh and 55% than Mo/Mo, for small sized carcinoma (0.1 cm) and approximately 50% than Rh/Rh and 125% than Mo/Mo, for big sized carcinoma (1 cm), considering 5 mGy X-ray beam. By decreasing lesion diameter and thickness, a limiting CT (100%) was occurred for size

  10. Development and evaluation of an indirect ELISA for detection of exfoliative toxin ExhA, ExhB or ExhC produced by Staphylococcus hyicus

    DEFF Research Database (Denmark)

    Andresen, Lars Ole

    1999-01-01

    Immunoblot analysis and enzyme-linked immunosorbent assay (ELISA) confirmed previous reports that the Staphylococcus hyicus exfoliative toxins ExhA and ExhB are metalloproteins, and further indicated that ExhC is also a metalloprotein. An indirect ELISA. was developed for the detection of toxigenic...... strains as an alternative method to the use of phage typing for selection of S. hyicus isolates to be used in autogenous vaccine against exudative epidermitis in pigs. The indirect ELISA was evaluated by investigating the presence of toxin among a total of 655 S. hyicus isolates from 69 pig skin samples......, one from each of the 69 pig herds with outbreak of exudative epidermitis. Toxigenic S. hyicus were detected in 74% of the cases by ELISA. From each of the five cases, in which initially no toxigenic S. hyicus were found, a further 40 S. hyicus-like colonies were tested in ELISA. Testing of this number...

  11. Enrichment and detection of microorganisms involved in direct and indirect methanogenesis from methanol in an anaerobic thermophilic bioreactor

    NARCIS (Netherlands)

    Roest, de K.; Altinbas, M.; Paulo, P.L.; Heilig, H.G.H.J.; Akkermans, A.D.L.; Smidt, H.; Vos, de W.M.; Stams, A.J.M.

    2005-01-01

    To gain insight into the microorganisms involved in direct and indirect methane formation from methanol in a laboratory-scale thermophilic (55°C) methanogenic bioreactor, reactor sludge was disrupted and serial dilutions were incubated in specific growth media containing methanol and possible

  12. Development & standardization of an in-house IgM indirect ELISA for the detection of parvovirus B19 infections

    Directory of Open Access Journals (Sweden)

    Kumaran Vadivel

    2017-01-01

    Interpretation & conclusions: The in-house IgM indirect ELISA was found to be simple with high sensitivity and specificity when compared with nPCR and could be used as an alternative to expensive commercial kits in resource-poor settings.

  13. Detection of Candida albicans Sap2 in cancer patient serum samples by an indirect competitive enzyme-linked immunosorbent assay for the diagnosis of candidiasis

    OpenAIRE

    Yicun Wang; Xiang Gao; J U Zhi gang; Jingyuan Liu; Shuai Dong; Li Wang

    2013-01-01

    Background: The secreted aspartyl proteinases 2 (Sap2) of Candida albicans (C. albicans) is a potential marker of candididasis. It is a virulence factor associated with adherence and tissue invasion. Aim: In order to detect Sap2 in clinical sera, we developed an indirect competitive enzyme-linked immunosorbent assay (ELISA). Materials and Methods: Polyclonal antibodies were produced for Sap2 by injecting Sap2 into a New Zealand White inbred rabbit. They could be used at a dilution exceeding 1...

  14. Inmunofluorescencia con Crithidia luciliae para la detección de anticuerpos anti-ADN: Imágenes atípicas y su relación con enfermedad de Chagas y leishmaniasis Immunofluorescence assay with Crithidia luciliae for the detection of anti-DNA antibodies: Atypical images and their relationship with Chagas’ disease and leishmaniasis

    Directory of Open Access Journals (Sweden)

    Gloria Griemberg

    2006-02-01

    Full Text Available Los anticuerpos anti-ADN nativo pueden detectarse por inmunofluorescencia indirecta con Crithidia luciliae, observándose tinción fluorescente anular del kinetoplasto que contiene ADN de doble cadena. En algunos casos pueden observarse imágenes fluorescentes en flagelo, membrana y corpúsculo basal, consideradas atípicas. Como C. luciliae pertenece a la familia Trypanosomatidae, que incluye patógenos para el hombre como Trypanosoma cruzi y Leishmaniaspp., se consideró que las imágenes atípicas pudieran deberse a reacciones cruzadas. Se realizaron estudios serológicos para Chagas a 105 muestras provenientes de zona endémica (Corrientes y no endémica (Buenos Aires para T. cruzi que presentaban imágenes atípicas con C. luciliae. La serología para Chagas resultó positiva en el 64.7% de las muestras de Buenos Aires y en el 78.3% de las de Corrientes que presentaban frente a C. luciliae imagen conjunta de membrana y flagelo. No presentaron la imagen conjunta ninguna de las muestras de dadores de sangre normales, ni de pacientes con enfermedades del tejido conectivo, excepto dos con lupus que también eran chagásicos. Todas las muestras de pacientes chagásicos analizadas frente a C. luciliae presentaron la imagen conjunta. Se estudiaron también 46 muestras de pacientes con leishmaniasis, 28 de ellos coinfectados con T. cruzi. La imagen conjunta se observó en el 88.0% de las muestras de leishmaniásicos y en el 78.5% de las de coinfectados. Los resultados sugieren que C. luciliae podría ser un sustrato alternativo, económico y de bajo riesgo para el diagnóstico serológico de enfermedad de Chagas, aunque no discrimina la infección por Leishmania. El hallazgo de la imagen conjunta en la detección de anti-ADN nativo señala la conveniencia de realizar en esos pacientes, estudios clínicos y de laboratorio para enfermedad de Chagas y leishmaniasis.Anti-native DNA antibodies can be detected by indirect immunofluorescence assay with

  15. Simultaneous determination of ammonia, dimethylamine, trimethylamine and trimethylamine-N-oxide in fish extracts by capillary electrophoresis with indirect UV-detection

    DEFF Research Database (Denmark)

    Timm Heinrich, Maike; Jørgensen, Bo

    2002-01-01

    A capillary electrophoretic method with indirect UV detection is described for simultaneous determination of ammonia, dimethylamine (DMA), trimethylamine (TMA) and trimethylamine-N- oxide (TMAO) in aqueous extracts of fish, A buffer consisting of 4 mM formic acid, 5 mM copper(II)sulfate and 3 m......M. The detection limit for ammonia, DMA, TMA, and TMAO was less than 0.04 mM, corresponding to 2 mg nitrogen per 100 g fish. As an extra benefit, the method also provided a quantitative determination of potassium, sodium, calcium and magnesium ions. (C) 2002 Elsevier Science Ltd. All rights reserved....

  16. Immunofluorescent determination of wheat protein in meat products

    Directory of Open Access Journals (Sweden)

    Michaela Petrášová

    2014-02-01

    foodstuffs is relatively difficult because of the fact that they occur in trace amounts and are often masked by various parts of the product. This paper deals with detection of wheat protein in meat products bought in the retail network of the Czech Republic. Ten cooked meat products, especially types of sausages and soft salami which stated wheat protein in their composition, were examined. The samples were processed using the method of immunofluorescence and stained with Texas Red fluorochrome. The presence of wheat protein was demonstrated in all the examined meat products. From the results it follows that the method of immunofluorescence is suitable for detection of wheat protein in meat products. Normal 0 21 false false false CS JA X-NONE

  17. Rapid Identification of Dengue Virus Serotypes Using Monoclonal Antibodies in an Indirect Immunofluorescence Test.

    Science.gov (United States)

    1982-06-18

    encephalitis(TBH-28), West Nile(E-101), Yellow fever(French neurotropic and 17D strains), and Zika . Two Sandfly Fever viruses (213452 and Candiru) were...were provided as first passage isolates ( Aedes pseudoscutellaris cells, AP-61) or human serum from recent dengue virus patients. African isolates... viruses of the Phlebovirus genus (Table 1). Several monoclonal antibody preparations reacted solely with dengue virus serotypes. Two preparations (13E7 and

  18. Quantitation of underivatized branched-chain amino acids in sport nutritional supplements by capillary electrophoresis with direct or indirect UV absorbance detection.

    Science.gov (United States)

    Qiu, Jun; Wang, Jinhao; Xu, Zhongqi; Liu, Huiqing; Ren, Jie

    2017-01-01

    The branched-chain amino acids (BCAAs) including leucine (Leu), isoleucine (Ile) and valine (Val) play a pivotal role in the human body. Herein, we developed capillary electrophoresis (CE) coupled with conventional UV detector to quantify underivatized BCAAs in two kinds of sport nutritional supplements. For direct UV detection at 195 nm, the BCAAs (Leu, two enantiomers of Ile and Val) were separated in a background electrolyte (BGE) consisting of 40.0 mmol/L sodium tetraborate, and 40.0 mmol/L β-cyclodextrin (β-CD) at pH 10.2. In addition, the indirect UV detection at 264 nm was achieved in a BGE of 2.0 mmol/L Na2HPO4, 10.0 mmol/L p-aminosalicylic acid (PAS) as UV absorbing probe, and 40.0 mmol/L β-CD at pH 12.2. The β-CD significantly benefited the isomeric separation of Leu, L- and D-Ile. The optimal conditions allowed the LODs (limit of detections) of direct and indirect UV absorption detection to be tens μmol/L level, which was comparable to the reported CE inline derivatization method. The RSDs (relative standard deviations) of migration time and peak area were less than 0.91% and 3.66% (n = 6). Finally, CE with indirect UV detection method was applied for the quantitation of BCAAs in two commercial sport nutritional supplements, and good recovery and precision were obtained. Such simple CE method without tedious derivatization process is feasible of quality control and efficacy evaluation of the supplemental proteins.

  19. Quantitation of underivatized branched-chain amino acids in sport nutritional supplements by capillary electrophoresis with direct or indirect UV absorbance detection.

    Directory of Open Access Journals (Sweden)

    Jun Qiu

    Full Text Available The branched-chain amino acids (BCAAs including leucine (Leu, isoleucine (Ile and valine (Val play a pivotal role in the human body. Herein, we developed capillary electrophoresis (CE coupled with conventional UV detector to quantify underivatized BCAAs in two kinds of sport nutritional supplements. For direct UV detection at 195 nm, the BCAAs (Leu, two enantiomers of Ile and Val were separated in a background electrolyte (BGE consisting of 40.0 mmol/L sodium tetraborate, and 40.0 mmol/L β-cyclodextrin (β-CD at pH 10.2. In addition, the indirect UV detection at 264 nm was achieved in a BGE of 2.0 mmol/L Na2HPO4, 10.0 mmol/L p-aminosalicylic acid (PAS as UV absorbing probe, and 40.0 mmol/L β-CD at pH 12.2. The β-CD significantly benefited the isomeric separation of Leu, L- and D-Ile. The optimal conditions allowed the LODs (limit of detections of direct and indirect UV absorption detection to be tens μmol/L level, which was comparable to the reported CE inline derivatization method. The RSDs (relative standard deviations of migration time and peak area were less than 0.91% and 3.66% (n = 6. Finally, CE with indirect UV detection method was applied for the quantitation of BCAAs in two commercial sport nutritional supplements, and good recovery and precision were obtained. Such simple CE method without tedious derivatization process is feasible of quality control and efficacy evaluation of the supplemental proteins.

  20. Detection of NP, N3 and N7 antibodies to avian influenza virus by indirect ELISA using yeast-expressed antigens

    Directory of Open Access Journals (Sweden)

    Ammayappan Arun

    2009-10-01

    Full Text Available Abstract Background Avian influenza viruses, belonging to the family Orthomyxoviridae, possess distinct combinations of hemagglutinin (H and the neuraminidase (N surface glycoproteins. Typing of both H and N antigens is essential for the epidemiological and surveillance studies. Therefore, it is important to find a rapid, sensitive, and specific method for their assay, and ELISA can be useful for this purpose, by using recombinant proteins. Results The nucleoprotein (NP and truncated neuraminidase subtype 3 and 7 of avian influenza virus (AIV were expressed in Saccharomyces cerevisiae and used to develop an indirect enzyme-linked immunosorbent assay for antibody detection. The developed assays were evaluated with a panel of 64 chicken serum samples. The performance of NP-ELISA was compared with the commercially available ProFlok® AIV ELISA kit. The results showed comparable agreement and sensitivity between the two tests, indicating that NP-ELISA assay can be used for screening the influenza type A antibody in AIV infected birds. The N3 and N7- ELISAs also reacted specifically to their type specific sera and did not exhibit any cross-reaction with heterologous neuraminidase subtype specific sera. Conclusion The study demonstrates the expression of the NP, N3, and N7 proteins of AIV in yeast (S. cerevisiae and their application in developing an indirect ELISA for detecting NP, N3 and N7 antibodies from AIV-infected chicken sera. The described indirect ELISAs are rapid, sensitive, specific and can be used as promising tests during serological surveillance.

  1. Immunofluorescent staining of nuclear antigen in lymphoid cells transformed by Herpesvirus papio (HVP).

    Science.gov (United States)

    Schmitz, H

    1981-01-01

    An improved fixation method for antigen detection in lymphoblastoid cells is described. Herpesvirus papio nuclear antigen (HUPNA) could be stained in several transformed lymphoid cell lines by anti-complement immunofluorescence (ACIF). Antibody to HUPNA was detected in many human sera containing antibodies to Epstein-Barr virus capsid and nuclear antigen (EBNA). Rheumatoid arthritis sera showed a high incidence of both anti-EBNA and anti-HUPNA antibodies.

  2. Tight connection between direct and indirect detection of dark matter through Higgs portal couplings to a hidden sector

    International Nuclear Information System (INIS)

    Arina, Chiara; Josse-Michaux, Francois-Xavier; Sahu, Narendra

    2010-01-01

    We present a hidden Abelian extension of the standard model including a complex scalar as a dark matter candidate and a light scalar acting as a long range force carrier between dark matter particles. The Sommerfeld enhanced annihilation cross section of the dark matter explains the observed cosmic ray excesses. The light scalar field also gives rise to potentially large cross sections of dark matter on the nucleon, therefore providing an interesting way to probe this model simultaneously at direct and indirect dark matter search experiments. We constrain the parameter space of the model by taking into account the CDMS-II exclusion limit as well as PAMELA and Fermi LAT data.

  3. Comparison of indirect hemagglutination and 51Chromium release tests for detection of herpes simplex virus types 1 and 2 antibodies in patients with recurrent herpes infections

    International Nuclear Information System (INIS)

    Kesavalu, L.; Seth, P.

    1980-01-01

    Indirect hemagglutination and 51 Cr release tests (IHAT and 51-CRT respectively) were compared in patients with recurrent herpes simplex virus (HSV) infections from whom HSV-1 or HSV-2 was isolated. Both tests were equally sensitive and specific in detecting HSV antibodies. However, IHAT was more specific in detecting homologous HSV antibody response in patients with recurrent HSV-2 infections. Past infections with HSV-1 in the patients with dual infections were detected by determining HSV-type specific antibodies by inhibition of IHAT. Cross absorption studies showed that the antibody reactivity measured by the two tests was qualitatively and quantitatively different. Nevertheless, IHAT has been found to be more appropriate test for seroepidemiologic studies of HSV-2 infections because of its specificity, rapidity and less cost, whereas, 51-CRT appears to measure antibodies against recent and more predominant type of infecting HSV. (Author)

  4. Direct immunofluorescence of normal skin in rheumatoid arthritis.

    Science.gov (United States)

    Fitzgerald, O M; Barnes, L; Woods, R; McHugh, L; Barry, C; O'Loughlin, S

    1985-11-01

    The clinical significance of previously described immunoglobulin and complement deposition in the superficial dermal vessel walls of patients with rheumatoid arthritis is unknown. In the present study, skin biopsies were obtained from the normal forearm and buttock of 48 unselected patients with rheumatoid arthritis and were examined by direct immunofluorescence (IF) for the presence of immunoglobulin (IgG,A,M) and complement (C3) in the vessel walls. Deposits of C3, IgM or IgG were detected in 10 patients. Five patients had deposits at the forearm sample alone, four patients had deposits at both biopsy sites, while one patient was positive at the buttock alone. Clinical features were similar in patients with and without vessel IF. However, patients with IF were significantly more seropositive with lower levels of complement and raised levels of serum IgA and IgM. There was also an increased level of circulating IgG immune complexes in these patients. Further analysis following exclusion of seronegative patients revealed similar results. This study suggests that the presence of vessel IF identifies a subgroup of patients who have evidence of more severe immunological disturbance.

  5. Anti-human neutrophil antigen-1a, -1b, and -2 antibodies in neonates and children with immune neutropenias analyzed by extracted granulocyte antigen immunofluorescence assay.

    Science.gov (United States)

    Onodera, Rie; Kurita, Emi; Taniguchi, Kikuyo; Karakawa, Shuhei; Okada, Satoshi; Kihara, Hirotaka; Fujii, Teruhisa; Kobayashi, Masao

    2017-11-01

    Anti-human neutrophil antigen (HNA) antibodies have been implicated in the development of neonatal alloimmune neutropenia (NAN) and autoimmune neutropenia (AIN). There are many conventional assay methods that detect anti-HNA antibodies. However, a method to measure multiple samples and detect several anti-HNA antibodies simultaneously is needed. We developed a new method, the extracted granulocyte antigen immunofluorescence assay (EGIFA), to analyze anti-HNA-1a, -1b, and -2 antibodies in sera. The results obtained by EGIFA were evaluated in comparison with those from several standard assay methods. Anti-HNA antibodies in serum samples from nine familial cases with suspected NAN (n = 19) and children with suspected AIN (n = 88) were also measured by EGIFA. The evaluation of nine serum samples with anti-HNA antibodies suggested that EGIFA demonstrated equivalent specificity and superior sensitivity to monoclonal antibody-specific immobilization of granulocyte antigens and had comparable sensitivity to the granulocyte indirect immunofluorescence test. EGIFA successfully detected anti-HNA-1a or -1b antibodies in seven of nine familial cases with suspected NAN. EGIFA detected anti-HNA antibodies in 40.9% of children with suspected AIN. Among them, isolated anti-HNA-1a or -1b antibody was detected in 4.5 or 12.5% of children, respectively, and anti-HNA-2 antibody was identified in 3.4% of children. The 30.8% (16 of 52) of children negative for anti-HNA antibody by EGIFA were positive for anti-HLA antibody. EGIFA facilitated the measurement of anti-HNA-1a, -1b, and/or -2 antibodies in sera. The prompt measurement of anti-HNA antibodies will improve the diagnosis and clinical management of patients with suspected NAN or AIN. © 2017 AABB.

  6. Surface-biofunctionalized multicore/shell CdTe@SiO2 composite particles for immunofluorescence assay

    Science.gov (United States)

    Jing, Lihong; Li, Yilin; Ding, Ke; Qiao, Ruirui; Rogach, Andrey L.; Gao, Mingyuan

    2011-12-01

    Strongly fluorescent multicore/shell structured CdTe@SiO2 composite particles of ~ 50 nm were synthesized via the reverse microemulsion method by using CdTe quantum dots co-stabilized by thioglycolic acid and thioglycerol. The optical stability of the CdTe@SiO2 composite particles in a wide pH range, under prolonged UV irradiation in pure water, or in different types of physiological buffers was systematically investigated. Towards immunofluorescence assay, both poly(ethylene glycol) (PEG) and carboxyl residues were simultaneously grafted on the surface of the silanol-terminated CdTe@SiO2 composite particles upon further reactions with silane reagents bearing a PEG segment and carboxyl group, respectively, in order to suppress the nonspecific interactions of the silica particles with proteins and meanwhile introduce reactive moieties to the fluorescent particles. Agarose gel electrophoresis, dynamic light scattering and conventional optical spectroscopy were combined to investigate the effectiveness of the surface modifications. Via the surface carboxyl residue, various antibodies were covalently conjugated to the fluorescent particles and the resultant fluorescent probes were used in detecting cancer cells through both direct fluorescent antibody and indirect fluorescent antibody assays, respectively.

  7. Diagnostic performance of CT and MRI on the detection of symptomatic intracranial dural arteriovenous fistula: a meta-analysis with indirect comparison

    Energy Technology Data Exchange (ETDEWEB)

    Lin, Yen-Heng [National Taiwan University, Institute of Epidemiology and Preventive Medicine, Taipei (China); National Taiwan University Hospital in Taipei and Yuan-Lin Branch, Department of Medical Imaging and Radiology, Hospital and Medical College, Taipei (China); Lin, Hsien-Ho [National Taiwan University, Institute of Epidemiology and Preventive Medicine, Taipei (China); Liu, Hon-Man; Lee, Chung-Wei; Chen, Ya-Fang [National Taiwan University Hospital in Taipei and Yuan-Lin Branch, Department of Medical Imaging and Radiology, Hospital and Medical College, Taipei (China)

    2016-08-15

    This study aims to review the diagnostic performance of computed tomography (CT) and magnetic resonance imaging (MRI) in symptomatic dural arteriovenous fistula (DAVF). EMBASE, PubMed, and Cochrane Library were searched until April 2015 for studies which compared CT, MRI, or both with angiography for the detection of DAVF. The diagnostic performances of MRI and CT were indirectly compared using modality as a covariate in the analysis. Thirteen studies met our inclusion criteria. MRI had a sensitivity of 0.90 (95 % confidence interval (CI) = 0.83-0.94) and specificity of 0.94 (95 % CI = 0.90-0.96). CT had a sensitivity of 0.80 (95 % CI = 0.62-0.90) and specificity of 0.87 (95 % CI = 0.74-0.94). MRI showed better diagnostic performance than CT (p = 0.02). Contrast medium use and time-resolved MR angiography did not improve MRI diagnostic performance (p = 0.31 and 0.44, respectively). Both CT and MRI had good diagnostic performance. MRI was better than CT on the detection of symptomatic intracranial dural arteriovenous fistula in the indirect comparison. (orig.)

  8. Capillary electrophoresis of chitooligosaccharides in acidic solution: simple determination using a quaternary-ammonium-modified column and indirect photometric detection with crystal violet.

    Science.gov (United States)

    Hattori, Toshiaki; Anraku, Nobuhiro; Kato, Ryo

    2010-02-01

    Five chitosan oligosaccharides were separated in acidic aqueous solution by capillary electrophoresis (CE) with indirect photometric detection using a positively coated capillary. Electrophoretic mobility of the chitooligosaccharides (COSs) depended on the number of monomer units in acidic aqueous solution, similar to other polyelectrolyte oligomers. The separation was developed in nitric acid aqueous solution at pH 3.0 with 1 mM Crystal Violet, using a capillary positively coated with N-trimethoxypropyl-N,N,N-trimethylammonium chloride. The limit of the detection for chitooligosaccharides with two to six saccharide chains was less than 5 microM. CE determination of an enzymatically hydrolyzed COS agreed with results from HPLC. 2009 Elsevier B.V. All rights reserved.

  9. Accuracy of the Enhanced Liver Fibrosis Test vs Fibrotest, Elastography and Indirect Markers in Detection of Advanced Fibrosis in Patients with Alcoholic Liver Disease

    DEFF Research Database (Denmark)

    Thiele, Maja; Madsen, Bjørn Stæhr; Hansen, Janne Fuglsang

    2018-01-01

    BACKGROUND & AIMS: Alcohol is the leading cause of cirrhosis and liver-related mortality, but we lack serum markers to detect compensated disease. We compared the accuracy of the Enhanced Liver Fibrosis test (ELF), the FibroTest, liver stiffness measurements (made by transient elastography and 2......-dimensional shear-wave elastography), and 6 indirect marker tests in detection of advanced liver fibrosis (Kleiner stage ≥F3). METHODS: We performed a prospective study of 10 liver fibrosis markers (patented and not), all performed on the same day. Patients were recruited from primary centers (municipal...... significantly from those of liver stiffness measurement in intention-to-diagnose analyses (AUROC for transient elastography, 0.90), but did differ in the per-protocol analysis (AUROC for transient elastography, 0.97) (P=.521 and .004 for comparison with ELF). Adding a serum marker to transient elastography...

  10. Indirect enzyme-linked immunosorbent assay method based on Streptococcus agalactiae rSip-Pgk-FbsA fusion protein for detection of bovine mastitis.

    Science.gov (United States)

    Bu, Ri-E; Wang, Jin-Liang; Wu, Jin-Hua; Xilin, Gao-Wa; Chen, Jin-Long; Wang, Hua

    2017-03-01

    The aim of this study was to establish a rapid and accurate method for the detection of the Streptococcus agalactiae antibody (SA-Ab) to determine the presence of the bovine mastitis (BM)-causative pathogen. The multi-subunit fusion protein rSip-Pgk-FbsA was prokaryotically expressed and purified. The triple activities of the membrane surface-associated proteins Sip, phosphoglycerate kinase (Pgk), and fibronectin (FbsA) were used as the diagnostic antigens to establish an indirect enzyme-linked immunosorbent assay (ELISA) method for the detection of SA-Ab in BM. The optimal antigen coating concentration was 2 μg/mL, the optimal serum dilution was 1:160, and the optimal dilution of the enzyme-labeled secondary antibody was 1:6000. The sensitivity, specificity, and repeatability tests showed that the method established in this study had no cross-reaction with antibodies to Streptococcus pyogenes, Escherichia coli, Staphylococcus aureus, and Staphylococcus epidermidis in the sera. The results of the sensitivity test showed that a positive result could be obtained even if the serum dilution reached 1:12,800, indicating the high sensitivity and good repeatability of the method. The positive coincidence rate of this method was 98.6%, which is higher than that of previous tests established with the Sip or Pgk mono-antigen fusion protein, respectively, demonstrating the relatively higher sensitivity of this newly established method. The detection rate for 389 clinical samples was 46.53%. The indirect ELISA method established in this study could provide a more accurate and reliable serological method for the rapid detection of S. agalactiae in cases of BM.

  11. Standardization of the indirect enzyme-linked immunosorbent assay for detection of antibodies against Newcastle disease virus in chickens

    International Nuclear Information System (INIS)

    Della Porta, A.J.; Young, J.; Hansson, E.; Spencer, T.

    1994-01-01

    Newcastle disease is the major viral disease of poultry causing significant economic losses in most countries except Australia and New Zealand. Serological monitoring of poultry has traditionally been carried out using the haemagglutinin-inhibition (HI) test. More recently, ELISA has been used for the same purpose. This paper described the use of an indirect ELISA for assay of antibodies in chickens against Newcastle disease viruses and compares some of the parameters for this test. The sucrose density gradient purified, inactivated, antigen enabled performance of the test without the addition of any blocking agents other than the usual Tween 20. A range of plates was compared and the most suitable plate was found to be a polystyrene haemagglutination plate giving an excellent positive to negative ratio of 33.2, compared with some expensive ELISA plates which gave very low +ve/-ve ratios. Various incubation conditions for the steps in the ELISA were compared and incubation with shaking at room temperature (24 to 28 deg. C) gave adequate reactivity whilst simplifying incubation conditions and speeding up the test. The negative cut-off value was determined by testing 1632 HI negative specific pathogen free sera from birds of a wide age range. The reactivity of sera in the ELISA was standardized using a standard curve on every plate and converting the readings to ELISA units (EUs) in the range of 16 to 512 EUs. The EU values of these sera were not normally distributed and so the 95% cut-off was determined by ranking the values in descending order and retaining only the top 5% of the values as false positives. This resulted in a cut-off value of 33.6 EUs, with few of HI positive sera having values lower than this cut-off. The use of a standard curve on each plate is recommended in order to standardize the assay and to determine the ELISA units for the test sera. (author). 14 refs, 2 figs, 1 tab

  12. Enrichment and detection of microorganisms involved in direct and indirect methanogenesis from methanol in an anaerobic thermophilic bioreactor.

    Science.gov (United States)

    Roest, Kees; Altinbas, Mahmut; Paulo, Paula L; Heilig, H G H J; Akkermans, Antoon D L; Smidt, Hauke; de Vos, Willem M; Stams, Alfons J M

    2005-10-01

    To gain insight into the microorganisms involved in direct and indirect methane formation from methanol in a laboratory-scale thermophilic (55 degrees C) methanogenic bioreactor, reactor sludge was disrupted and serial dilutions were incubated in specific growth media containing methanol and possible intermediates of methanol degradation as substrates. With methanol, growth was observed up to a dilution of 10(8). However, when Methanothermobacter thermoautotrophicus strain Z245 was added for H2 removal, growth was observed up to a 10(10)-fold dilution. With H2/CO2 and acetate, growth was observed up to dilutions of 10(9) and 10(4), respectively. Dominant microorganisms in the different dilutions were identified by 16S rRNA-gene diversity and sequence analysis. Furthermore, dilution polymerase chain reaction (PCR) revealed a similar relative abundance of Archaea and Bacteria in all investigated samples, except in enrichment with acetate, which contained 100 times less archaeal DNA than bacterial DNA. The most abundant bacteria in the culture with methanol and strain Z245 were most closely related to Moorella glycerini. Thermodesulfovibrio relatives were found with high sequence similarity in the H2/CO2 enrichment, but also in the original laboratory-scale bioreactor sludge. Methanothermobacter thermoautotrophicus strains were the most abundant hydrogenotrophic archaea in the H2/CO2 enrichment. The dominant methanol-utilizing methanogen, which was present in the 10(8)-dilution, was most closely related to Methanomethylovorans hollandica. Compared to direct methanogenesis, results of this study indicate that syntrophic, interspecies hydrogen transfer-dependent methanol conversion is equally important in the thermophilic bioreactor, confirming previous findings with labeled substrates and specific inhibitors.

  13. Thin film organic photodetectors for indirect X-ray detection demonstrating low dose rate sensitivity at low voltage operation

    Energy Technology Data Exchange (ETDEWEB)

    Starkenburg, Daken J. [Department of Materials Science and Engineering, University of Florida, Gainesville, Florida 32611, USA; Johns, Paul M. [Nuclear Engineering Program, University of Florida, Gainesville, Florida 32611, USA; Detection Systems Group, Pacific Northwest National Laboratory, Richland, Washington 99354, USA; Baciak, James E. [Department of Materials Science and Engineering, University of Florida, Gainesville, Florida 32611, USA; Nuclear Engineering Program, University of Florida, Gainesville, Florida 32611, USA; Nino, Juan C. [Department of Materials Science and Engineering, University of Florida, Gainesville, Florida 32611, USA; Xue, Jiangeng [Department of Materials Science and Engineering, University of Florida, Gainesville, Florida 32611, USA

    2017-12-14

    Developments in the field of organic semiconductors have generated organic photodetectors with high quantum efficiency, wide spectral sensitivity, low power consumption, and unique form factors that are flexible and conformable to their substrate shape. In this work, organic photodetectors coupled with inorganic CsI(Tl) scintillators are used to showcase the low dose rate sensitivity that is enabled when high performance organic photodetectors and scintillator crystals are integrated. The detection capability of these organic-inorganic coupled systems to high energy radiation highlights their potential as an alternative to traditional photomultiplier tubes for nuclear spectroscopy applications. When exposed to Bremsstrahlung radiation produced from an X-ray generator, SubPc:C60, AlPcCl:C70, and P3HT:PC61BM thin film photodetectors with active layer thicknesses less than 100 nm show detection of incident radiation at low and no applied bias. Remarkably low dose rates, down to at least 0.28 µGy/s, were detectable with a characteristic linear relationship between exposure rate and photodetector current output. These devices also demonstrate sensitivities as high as 5.37 mC Gy-1 cm-2 when coupled to CsI(Tl). Additionally, as the tube voltage across the X-ray generator was varied, these organic-inorganic systems showed their ability to detect a range of continuous radiation spectra spanning several hundred keV.

  14. Capability of the HAWC Gamma-Ray Observatory for the Indirect Detection of Ultrahigh-Energy Neutrinos

    Directory of Open Access Journals (Sweden)

    Hermes León Vargas

    2017-01-01

    Full Text Available The detection of ultrahigh-energy neutrinos, with energies in the PeV range or above, is a topic of great interest in modern astroparticle physics. The importance comes from the fact that these neutrinos point back to the most energetic particle accelerators in the Universe and provide information about their underlying acceleration mechanisms. Atmospheric neutrinos are a background for these challenging measurements, but their rate is expected to be negligible above ≈1 PeV. In this work we describe the feasibility to study ultrahigh-energy neutrinos based on the Earth-skimming technique, by detecting the charged leptons produced in neutrino-nucleon interactions in a high mass target. We propose to detect the charged leptons, or their decay products, with the High Altitude Water Cherenkov (HAWC observatory and use as a large-mass target for the neutrino interactions the Pico de Orizaba volcano, the highest mountain in Mexico. In this work we develop an estimate of the detection rate using a geometrical model to calculate the effective area of the observatory. Our results show that it may be feasible to perform measurements of the ultrahigh-energy neutrino flux from cosmic origin during the expected lifetime of the HAWC observatory.

  15. Thin film organic photodetectors for indirect X-ray detection demonstrating low dose rate sensitivity at low voltage operation

    Science.gov (United States)

    Starkenburg, Daken J.; Johns, Paul M.; Baciak, James E.; Nino, Juan C.; Xue, Jiangeng

    2017-12-01

    Developments in the field of organic semiconductors have generated organic photodetectors with high quantum efficiency, wide spectral sensitivity, low power consumption, and unique form factors that are flexible and conformable to their substrate shape. In this work, organic photodetectors coupled with inorganic CsI(Tl) scintillators are used to showcase the low dose rate sensitivity that is enabled when high performance organic photodetectors and scintillator crystals are integrated. The detection capability of these organic-inorganic coupled systems to high energy radiation highlights their potential as an alternative to traditional photomultiplier tubes for nuclear spectroscopy applications. When exposed to Bremsstrahlung radiation produced from an X-ray generator, SubPc:C60, AlPcCl:C70, and P3HT:PC61BM thin film photodetectors with active layer thicknesses less than 100 nm show detection of incident radiation at low and no applied bias. Remarkably low dose rates, down to at least 0.18 μGy/s, were detectable with a characteristic linear relationship between exposure rate and photodetector current output. These devices also demonstrate sensitivities as high as 5.37 mC Gy-1 cm-2 when coupled to CsI(Tl). Additionally, as the tube voltage across the X-ray generator was varied, these organic-inorganic systems showed their ability to detect a range of continuous radiation spectra spanning several hundred keV.

  16. Rapid Detection and Identification of Streptococcus Iniae Using a Monoclonal Antibody-Based Indirect Fluorescent Antibody Technique

    Science.gov (United States)

    Streptococcus iniae is among the major pathogens of a large number of fish species cultured in fresh and marine recirculating and net pen production systems . The traditional plate culture technique to detect and identify S. iniae is time consuming and may be problematic due to phenotypic variations...

  17. Direct and indirect singlet scalar dark matter detection in the lepton-specific two-Higgs-doublet model

    International Nuclear Information System (INIS)

    Boucenna, M. S.; Profumo, S.

    2011-01-01

    A recent study of gamma-ray data from the Galactic center motivates the investigation of light (∼7-10 GeV) particle dark matter models featuring tau-lepton pairs as dominant annihilation final state. The lepton-specific two-Higgs-doublet model provides a natural framework where light, singlet scalar dark matter can pair-annihilate dominantly into tau leptons. We calculate the nucleon-dark matter cross section for singlet scalar dark matter within the lepton-specific two-Higgs-doublet model framework, and compare with recent results from direct detection experiments. We study how direct dark matter searches can be used to constrain the dark matter interpretation of gamma-ray observations, for different dominant annihilation final states. We show that models exist with the correct thermal relic abundance that could fit the claimed gamma-ray excess from the Galactic center region and have direct detection cross sections of the order of what is needed to interpret recent anomalous events reported by direct detection experiments.

  18. Astrophysical limitations to the identification of dark matter: Indirect neutrino signals vis-a-vis direct detection recoil rates

    International Nuclear Information System (INIS)

    Serpico, Pasquale D.; Bertone, Gianfranco

    2010-01-01

    A convincing identification of dark matter (DM) particles can probably be achieved only through a combined analysis of different detections strategies, which provides an effective way of removing degeneracies in the parameter space of DM models. In practice, however, this program is made complicated by the fact that different strategies depend on different physical quantities, or on the same quantities but in a different way, making the treatment of systematic errors rather tricky. We discuss here the uncertainties on the recoil rate in direct-detection experiments and on the muon rate induced by neutrinos from dark matter annihilations in the Sun, and we show that, contrarily to the local DM density or overall cross section scale, irreducible astrophysical uncertainties affect the two rates in a different fashion, therefore limiting our ability to reconstruct the parameters of the dark matter particles. By varying within their respective errors astrophysical parameters such as the escape velocity and the velocity dispersion of dark matter particles, we show that the uncertainty on the relative strength of the neutrino and direct-detection signal is as large as a factor of 2 for typical values of the parameters, but can be even larger in some circumstances.

  19. Direct and indirect signal detection of 122 keV photons with a novel detector combining a pnCCD and a CsI(Tl) scintillator

    Energy Technology Data Exchange (ETDEWEB)

    Schlosser, D.M., E-mail: dieter.schlosser@pnsensor.de [PNSensor GmbH, Sckellstraße 3, 81667 München (Germany); Huth, M.; Hartmann, R. [PNSensor GmbH, Sckellstraße 3, 81667 München (Germany); Abboud, A.; Send, S. [Universität Siegen, Walter-Flex-Straße 3, 57072 Siegen (Germany); Conka-Nurdan, T. [Türkisch-Deutsche Universität, Sakinkaya Cad. 86, Beykoz, 34820 Istanbul (Turkey); Shokr, M.; Pietsch, U. [Universität Siegen, Walter-Flex-Straße 3, 57072 Siegen (Germany); Strüder, L. [PNSensor GmbH, Sckellstraße 3, 81667 München (Germany); Universität Siegen, Walter-Flex-Straße 3, 57072 Siegen (Germany)

    2016-01-01

    By combining a low noise fully depleted pnCCD detector with a CsI(Tl) scintillator, an energy-dispersive area detector can be realized with a high quantum efficiency (QE) in the range from below 1 keV to above 100 keV. In direct detection mode the pnCCD exhibits a relative energy resolution of 1% at 122 keV and spatial resolution of less than 75 µm, the pixel size of the pnCCD. In the indirect detection mode, i.e. conversion of the incoming X-rays in the scintillator, the measured energy resolution was about 9–13% at 122 keV, depending on the depth of interaction in the scintillator, while the position resolution, extracted with the help of simulations, was 30 µm only. We show simulated data for incident photons of 122 keV and compare the various interaction processes and relevant physical parameters to experimental results obtained with a radioactive {sup 57}Co source. - Highlights: • Position and energy resolving pnCCD+CsI(Tl) detector for energies from 1-150 keV • Detection in the pnCCD (122keV): 1% energy and <75µm spatial resolution • Detection in the scintillator (122keV): 9-12% energy and ~30µm spatial resolution.

  20. Establishment of an indirect solid phase radioimmunoassay for the detection of HSV-type I-specific antibodies in human serum

    International Nuclear Information System (INIS)

    Buchow, H.

    1982-01-01

    An indirect solid phase radioimmunoassay (IFRIA) was developed for the detection in human serum of antibodies against Herpes simplex virus (HSV) type I. The IFRIA was carried out according to the 'sandwich' principle. In comparison to other serological routine methods, especially the complement binding reaction (CBR) commonly used for HSV diagnostic, the IFRIA excelled by being a test which is quick to carry out and by lacking the subjectivity by the researcher in the evaluation of the test results, as well as by the use of patients serum which had not been previously treated. Comparative studies between IFRIA and CBR resulted in no agreement between the respectively attained maximum titer. As a reason for this the different qualities of the antibodies which can be determined by each test were discussed. For the addition of this test to the routine diagnostic further studies in the preservation of the specificity of the antigen in the case of longer storage are requisite. (orig.) [de

  1. Measurement of 14N quadrupole couplings in biomolecular solids using indirect-detection 14N solid-state NMR with DNP.

    Science.gov (United States)

    Jarvis, J A; Haies, I; Lelli, M; Rossini, A J; Kuprov, I; Carravetta, M; Williamson, P T F

    2017-11-07

    The quadrupolar interaction experienced by the spin-1 14 N nucleus is known to be extremely sensitive to local structure and dynamics. Furthermore, the 14 N isotope is 99.6% naturally abundant, making it an attractive target for characterisation of nitrogen-rich biological molecules by solid-state NMR. In this study, dynamic nuclear polarization (DNP) is used in conjunction with indirect 14 N detected solid-state NMR experiments to simultaneously characterise the quadrupolar interaction at multiple 14 N sites in the backbone of the microcrystalline protein, GB3. Considerable variation in the quadrupolar interaction (>700 kHz) is observed throughout the protein backbone. The distribution in quadrupolar interactions observed reports on the variation in local backbone conformation and subtle differences in hydrogen-bonding; demonstrating a new route to the structural and dynamic analysis of biomolecules.

  2. Application of the indirect enzyme-labeled antibody microtest to the detection and surveillance of animal diseases. [Brucellosis, cholera, and trichinosis in cattle and swine

    Energy Technology Data Exchange (ETDEWEB)

    Saunders, G.C. Clinard, E.H.; Bartlett, M.L.; Sanders, W.M.

    1976-01-01

    The rapid, indirect enzyme-labeled antibody (ELA) microplate test has been developed as a diagnostic and surveillance tool to aid in the control of animal disease. Data are presented, which illustrate the application of the test to viral (hog cholera), parasitic (trichinosis), and bacterial (brucellosis) diseases of animals. A greater than 95 percent correlation was observed between the hog cholera ELA test and the hog cholera serum neutralization test performed on over 2000 mixed hog cholera positive and negative field samples obtained during the 1976 New Jersey epizootic. Of 56 swine naturally infected with Trichinella spiralis at a level considered dangerous to man, all were ELA positive, while only one of 360 T. spiralis negative packing house sera was ELA positive. Preliminary experiments with bovine brucellosis (Brucella abortus) indicate that the ELA test is more sensitive than other test methods currently in use. ELA procedures should soon become tests of choice for the detection of antibodies to animal disease agents.

  3. Role of direct immunofluorescence in the diagnosis of glomerulonephritis

    Directory of Open Access Journals (Sweden)

    Archana C Buch

    2015-01-01

    Full Text Available Background: Immunofluorescence microscopy is a vital tool for the diagnosis of glomerular diseases. This study was carried out to study patterns of glomerulonephritis (GN and to record the sensitivity of direct immunofluorescence (DIF in renal lesions. The DIF findings were correlated with clinical and histopathology findings and discrepancies were analyzed. Materials and Methods: The cross-sectional analytical study was conducted during the period July 2011 to July 2013 at a tertiary care Hospital, Department of Pathology. A total of 75 renal biopsies were received for routine and immunofluorescence studies in which histopathology and clinical data were reviewed and analyzed. Results: The sensitivity of DIF was 87.9% and specificity was 70.5%. The maximum number of cases were seen in the age group 41-50 years. The pattern of GN by DIF was minimal change disease (MCD in 24%, IgA nephropathy in 13%, focal segmental glomerulosclerosis in 9% and membranoproliferative glomerulonephritis in 8% of the cases. Twelve histopathologically proven cases of GN were negative on DIF. One case of MCD on histopathology was diagnosed as IgM nephropathy based on DIF. Conclusion: Direct immunofluorescence forms an important diagnostic tool in reaching the exact diagnosis in various types of GN presenting with overlapping clinical and histomorphological features.

  4. Indirect ELISA based on Hendra and Nipah virus proteins for the detection of henipavirus specific antibodies in pigs.

    Directory of Open Access Journals (Sweden)

    Kerstin Fischer

    Full Text Available Hendra virus (HeV and Nipah virus (NiV belong to the genus Henipavirus in the family Paramyxoviridae. Henipavirus infections were first reported in the 1990's causing severe and often fatal outbreaks in domestic animals and humans in Southeast Asia and Australia. NiV infections were observed in humans in Bangladesh, India and in the first outbreak in Malaysia, where pigs were also infected. HeV infections occurred in horses in the North-Eastern regions of Australia, with singular transmission events to humans. Bats of the genus Pteropus have been identified as the reservoir hosts for henipaviruses. Molecular and serological indications for the presence of henipa-like viruses in African fruit bats, pigs and humans have been published recently. In our study, truncated forms of HeV and NiV attachment (G proteins as well as the full-length NiV nucleocapsid (N protein were expressed using different expression systems. Based on these recombinant proteins, Enzyme-linked Immunosorbent Assays (ELISA were developed for the detection of HeV or NiV specific antibodies in porcine serum samples. We used the NiV N ELISA for initial serum screening considering the general reactivity against henipaviruses. The G protein based ELISAs enabled the differentiation between HeV and NiV infections, since as expected, the sera displayed higher reactivity with the respective homologous antigens. In the future, these assays will present valuable tools for serosurveillance of swine and possibly other livestock or wildlife species in affected areas. Such studies will help assessing the potential risk for human and animal health worldwide by elucidating the distribution of henipaviruses.

  5. EUROPattern Suite technology for computer-aided immunofluorescence microscopy in autoantibody diagnostics.

    Science.gov (United States)

    Krause, C; Ens, K; Fechner, K; Voigt, J; Fraune, J; Rohwäder, E; Hahn, M; Danckwardt, M; Feirer, C; Barth, E; Martinetz, T; Stöcker, W

    2015-04-01

    Antinuclear autoantibodies (ANA) are highly informative biomarkers in autoimmune diagnostics. The increasing demand for effective test systems, however, has led to the development of a confusingly large variety of different platforms. One of them, the indirect immunofluorescence (IIF), is regarded as the common gold standard for ANA screening, as described in a position statement by the American College of Rheumatology in 2009. Technological solutions have been developed aimed at standardization and automation of IIF to overcome methodological limitations and subjective bias in IIF interpretation. In this review, we present the EUROPattern Suite, a system for computer-aided immunofluorescence microscopy (CAIFM) including automated acquisition of digital images and evaluation of IIF results. The system was originally designed for ANA diagnostics on human epithelial cells, but its applications have been extended with the latest system update version 1.5 to the analysis of antineutrophil cytoplasmic antibodies (ANCA) and anti-dsDNA antibodies. © The Author(s) 2015 Reprints and permissions: sagepub.co.uk/journalsPermissions.nav.

  6. Effects of blood contamination of cerebrospinal fluid on results of indirect fluorescent antibody tests for detection of antibodies against Sarcocystis neurona and Neospora hughesi.

    Science.gov (United States)

    Finno, Carrie J; Packham, Andrea E; David Wilson, W; Gardner, Ian A; Conrad, Patricia A; Pusterla, Nicola

    2007-05-01

    The purpose of this study was to determine the effect of blood contamination of cerebrospinal fluid (CSF) on the results of indirect fluorescent antibody tests (IFATs) for Sarcocystis neurona and Neospora hughesi. The in vitro study used antibody-negative CSF collected from non-neurologic horses immediately after euthanasia and blood samples from 40 healthy horses that had a range of IFAT antibody titers against S. neurona and N. hughesi. Serial dilutions of whole blood were made in seronegative CSF to generate blood-contaminated CSF with red blood cell (RBC) concentrations ranging from 10 to 100,000 RBCs/microl. The blood-contaminated CSF samples were then tested for antibodies against both pathogens using IFAT. Blood contamination of CSF had no detectable effect on IFAT results for S. neurona or N. hughesi at any serologic titer when the RBC concentration in CSF was or=5) for S. neurona and N. hughesi were detected only when the corresponding serum titers were >or=160 and >or=80, respectively. The IFAT performed on CSF is reliable for testing horses for equine protozoal myeloencephalitis caused by S. neurona or N. hughesi, even when blood contamination causes the RBC concentration in CSF to be up to 10,000 RBCs/microl.

  7. Development and evaluation of a truncated recombinant NS3 antigen-based indirect ELISA for detection of pestivirus antibodies in sheep and goats.

    Science.gov (United States)

    Kalaiyarasu, Semmannan; Mishra, Niranjan; Rajukumar, Katherukamem; Nema, Ram Kumar; Behera, Sthita Pragnya

    2015-01-01

    The aim of this study was to develop an indirect ELISA using the helicase domain of bovine viral diarrhoea virus (BVDV) NS3 protein instead of full-length NS3 protein for detection of BVDV and BDV antibodies in sheep and goats and its validation by comparing its sensitivity and specificity with virus neutralization test (VNT) as the reference test. The purified 50 kDa recombinant NS3 protein was used as the coating antigen in the ELISA. The optimal concentration of antigen was 320 ng/well at a serum dilution of 1:20 and the optimal positive cut-off optical density value was 0.40 based on test results of 418 VNT negative sheep and goat sera samples. When 569 serum samples from sheep (463) and goats (106) were tested, the ELISA showed a sensitivity of 91.71% and specificity of 94.59% with BVDV VNT. A good correlation (93.67%) was observed between the two tests. It showed a sensitivity of 85% and specificity of 86.6% with VNT in detecting BDV antibody positive or negative samples. This study demonstrates the efficacy of truncated recombinant NS3 antigen based ELISA for seroepidemiological study of pestivirus infection in sheep and goats.

  8. Comparison of Rose Bengal Plate Agglutination, Standard tube agglutination and Indirect ELISA tests for detection of Brucella antibodies in Cows and Buffaloes

    Directory of Open Access Journals (Sweden)

    S. N. Ghodasara

    2010-04-01

    Full Text Available A total of 180 serum samples (107 cows, 73 buffaloes from cases of abortion and various reproductive disorders were collected for detection of Brucella antibody by Rose Bengal Plate Agglutination Test (RBPT, Serum Tube Agglutination Test (STAT and indirect- ELISA (i-ELISA. The overall prevalence of brucellosis by RBPT, STAT and i-ELISA were 11.21%, 16.00% and 24.30% in cows 9.59%, 12.33% and 26.03% in buffaloes respectively. Overall seroprevalence of Brucellosis in cases of abortion, R.O.P. by RBPT, STAT and i-ELISA were 11.32%, 16.04% and 32.08% respectively. When three serological tests were compared, seropositivity was found highest by i-ELISA (25%, followed by STAT (14.45% and RBPT (10.56%. The results shows higher prevalence of brucellosis in cases of abortion and R.O.P., while at lower level from various reproductive disorders as detected serologically indicating endemicity of the infection in villages around Anand city, Gujarat. [Vet. World 2010; 3(2.000: 61-64

  9. Rapid antigen detection test for respiratory syncytial virus diagnosis as a diagnostic tool.

    Science.gov (United States)

    Mesquita, Flávio da Silva; Oliveira, Danielle Bruna Leal de; Crema, Daniela; Pinez, Célia Miranda Nunes; Colmanetti, Thaís Cristina; Thomazelli, Luciano Matsumia; Gilio, Alfredo Elias; Vieira, Sandra Elisabeth; Martinez, Marina Baquerizo; Botosso, Viviane Fongaro; Durigon, Edison Luiz

    The aim of this study was to evaluate the QuickVue ® RSV Test Kit (QUIDEL Corp, CA, USA) as a screening tool for respiratory syncytial virus in children with acute respiratory disease in comparison with the indirect immunofluorescence assay as gold standard. In Brazil, rapid antigen detection tests for respiratory syncytial virus are not routinely utilized as a diagnostic tool, except for the diagnosis of dengue and influenza. The authors retrospectively analyzed 486 nasopharyngeal aspirate samples from children under age 5 with acute respiratory infection, between December 2013 and August 2014, the samples were analyzed by indirect immunofluorescence assay and QuickVue ® RSV Test kit. Samples with discordant results were analyzed by real time PCR and nucleotide sequencing. From 313 positive samples by immunofluorescence assays, 282 (90%) were also positive by the rapid antigen detection test, two were positive only by rapid antigen detection test, 33 were positive only by immunofluorescence assays, and 171 were positive by both methods. The 35 samples with discordant results were analyzed by real time PCR; the two samples positive only by rapid antigen detection test and the five positive only by immunofluorescence assays were also positive by real time PCR. There was no relation between the negativity by QuickVue ® RSV Test and viral load or specific strain. The QuickVue ® RSV Test showed sensitivity of 90%, specificity of 98.8%, predictive positive value of 99.3%, and negative predictive value of 94.6%, with accuracy of 93.2% and agreement κ index of 0.85 in comparison to immunofluorescence assay. This study demonstrated that the QuickVue ® RSV Test Kit can be effective in early detection of Respiratory syncytial virus in nasopharyngeal aspirate and is reliable for use as a diagnostic tool in pediatrics. Copyright © 2016 Sociedade Brasileira de Pediatria. Published by Elsevier Editora Ltda. All rights reserved.

  10. Rapid antigen detection test for respiratory syncytial virus diagnosis as a diagnostic tool,

    Directory of Open Access Journals (Sweden)

    Flávio da Silva Mesquita

    Full Text Available Abstract Objective: The aim of this study was to evaluate the QuickVue® RSV Test Kit (QUIDEL Corp, CA, USA as a screening tool for respiratory syncytial virus in children with acute respiratory disease in comparison with the indirect immunofluorescence assay as gold standard. In Brazil, rapid antigen detection tests for respiratory syncytial virus are not routinely utilized as a diagnostic tool, except for the diagnosis of dengue and influenza. Methods: The authors retrospectively analyzed 486 nasopharyngeal aspirate samples from children under age 5 with acute respiratory infection, between December 2013 and August 2014, the samples were analyzed by indirect immunofluorescence assay and QuickVue® RSV Test kit. Samples with discordant results were analyzed by real time PCR and nucleotide sequencing. Results: From 313 positive samples by immunofluorescence assays, 282 (90% were also positive by the rapid antigen detection test, two were positive only by rapid antigen detection test, 33 were positive only by immunofluorescence assays, and 171 were positive by both methods. The 35 samples with discordant results were analyzed by real time PCR; the two samples positive only by rapid antigen detection test and the five positive only by immunofluorescence assays were also positive by real time PCR. There was no relation between the negativity by QuickVue® RSV Test and viral load or specific strain. The QuickVue® RSV Test showed sensitivity of 90%, specificity of 98.8%, predictive positive value of 99.3%, and negative predictive value of 94.6%, with accuracy of 93.2% and agreement κ index of 0.85 in comparison to immunofluorescence assay. Conclusions: This study demonstrated that the QuickVue® RSV Test Kit can be effective in early detection of Respiratory syncytial virus in nasopharyngeal aspirate and is reliable for use as a diagnostic tool in pediatrics.

  11. USE OF AN INDIRECT ENZYME-LINKED IMMUNOSORBENT ASSAY (ELISA TO DETECT ANTIBODIES IN AYU (Plecogiossus altivelis VACCINATED BY IMMERSION ADMINISTRATION

    Directory of Open Access Journals (Sweden)

    . Sukenda

    2007-05-01

    Full Text Available ABSTRACTAn indirect enzyme-linked immunosorbent assay (ELISA was used to detect serum antibody in ayu, Plecoglossus altivelis, immunized against Pseudomonasplecoglossicida by immersion vaccination.  First, the procedure of the ELISA was optimized and the sensitivity was checked.  Secondly, the formalin-killed cells (FKC of P. plecoglossicida was administered to ayu by immersion vaccination.  Two weeks after vaccination, fish were divided into two groups, one group was given booster.  The level of specific antibody production of both boostered and vaccinated only fish were statistically higher than unvaccinated control fish at the time of each blood collection.  However, the differences between the boostered and vaccinated only fish were not statistically significant.Keywords :  immunization, Pseudomonas plecoglossicida, ayu, ELISA ABSTRAKIndirect enzyme-linked immunosorbent assay (ELISA digunakan untuk mendeteksi antibodi pada ayu, Plecoglossus altivelis, yang diimunisasi dengan cara perendaman untuk melawan infeksi Pseudomonas plecoglossicida.  Pertama, prosedur ELISA dioptimasikan dan sensitivitas dari metode ini juga diperiksa.  Kemudian, bakteri Plecoglossus altivelis yang sudah dimatikan dengan formalin diberikan ke ikan ayu dengan vaksinasi perendaman.  Dua minggu setelah vaksinasi, ikan dibagi menjadi dua kelompok, satu kelompok diberi vaksinasi kedua.  Produksi antibodi spesifik dari ikan-ikan yang divaksinasi satu kali dengan vaksinasi dua kaii secara statistik lebih tinggi dibandingkan dengan control.  Akan tetapi, tidak ada perbedaan produksi antibodi antara ikan yarig divaksanisi satu kali dengan divaksinasi dua kali.Kata kunci :  imunisasi, Pseudomonasplecoglossicida, ayu, ELISA

  12. Development of Indirect Competitive Immuno-Assay Method Using SPR Detection for Rapid and Highly Sensitive Measurement of Salivary Cortisol Levels

    International Nuclear Information System (INIS)

    Tahara, Yusuke; Huang, Zhe; Kiritoshi, Tetsuro; Onodera, Takeshi; Toko, Kiyoshi

    2014-01-01

    The monitoring of salivary cortisol as a key biomarker of an individual’s stress response has been increasingly focused on. This paper describes the development of a novel cortisol immuno-assay method based on an indirect competitive method using a commercially available surface plasmon resonance instrument. The surface of an Au chip was modified with PEG6-COOH aromatic dialkanethiol self-assembled monolayers and hydrocortisone 3-(O-carboxymethyl) oxime (hydrocortisone 3-CMO) as a cortisol analog. A detection limit of 38 ppt range with a measurement range of 10 ppt–100 ppb was accomplished without the incubation of a mixing solution consisting of standard cortisol and an anti-cortisol antibody, and the time for quantification of cortisol concentration was 8 min from the sample injection. We experimentally compared our immuno-assay with a commercialized salivary cortisol enzyme-linked immunosorbent assay (ELISA) kit using human saliva samples. It was found that the results obtained by the cortisol immuno-assay had a good correlation with those obtained by ELISA assay (R = 0.96). Our findings indicate the potential utility of the cortisol immuno-assay for measurements of human salivary cortisol levels.

  13. Development of Indirect Competitive Immuno-Assay Method Using SPR Detection for Rapid and Highly Sensitive Measurement of Salivary Cortisol Levels

    Energy Technology Data Exchange (ETDEWEB)

    Tahara, Yusuke; Huang, Zhe; Kiritoshi, Tetsuro [Graduate School of Information Science and Electrical Engineering, Kyushu University, Fukuoka (Japan); Onodera, Takeshi [Research and Development Center for Taste and Odor Sensing, Kyushu University, Fukuoka (Japan); Toko, Kiyoshi, E-mail: toko@ed.kyushu-u.ac.jp [Graduate School of Information Science and Electrical Engineering, Kyushu University, Fukuoka (Japan); Research and Development Center for Taste and Odor Sensing, Kyushu University, Fukuoka (Japan)

    2014-05-30

    The monitoring of salivary cortisol as a key biomarker of an individual’s stress response has been increasingly focused on. This paper describes the development of a novel cortisol immuno-assay method based on an indirect competitive method using a commercially available surface plasmon resonance instrument. The surface of an Au chip was modified with PEG6-COOH aromatic dialkanethiol self-assembled monolayers and hydrocortisone 3-(O-carboxymethyl) oxime (hydrocortisone 3-CMO) as a cortisol analog. A detection limit of 38 ppt range with a measurement range of 10 ppt–100 ppb was accomplished without the incubation of a mixing solution consisting of standard cortisol and an anti-cortisol antibody, and the time for quantification of cortisol concentration was 8 min from the sample injection. We experimentally compared our immuno-assay with a commercialized salivary cortisol enzyme-linked immunosorbent assay (ELISA) kit using human saliva samples. It was found that the results obtained by the cortisol immuno-assay had a good correlation with those obtained by ELISA assay (R = 0.96). Our findings indicate the potential utility of the cortisol immuno-assay for measurements of human salivary cortisol levels.

  14. Identification of inorganic improvised explosive devices by analysis of postblast residues using portable capillary electrophoresis instrumentation and indirect photometric detection with a light-emitting diode.

    Science.gov (United States)

    Hutchinson, Joseph P; Evenhuis, Christopher J; Johns, Cameron; Kazarian, Artaches A; Breadmore, Michael C; Macka, Miroslav; Hilder, Emily F; Guijt, Rosanne M; Dicinoski, Greg W; Haddad, Paul R

    2007-09-15

    A commercial portable capillary electrophoresis (CE) instrument has been used to separate inorganic anions and cations found in postblast residues from improvised explosive devices (IEDs) of the type used frequently in terrorism attacks. The purpose of this analysis was to identify the type of explosive used. The CE instrument was modified for use with an in-house miniaturized light-emitting diode (LED) detector to enable sensitive indirect photometric detection to be employed for the detection of 15 anions (acetate, benzoate, carbonate, chlorate, chloride, chlorite, cyanate, fluoride, nitrate, nitrite, perchlorate, phosphate, sulfate, thiocyanate, thiosulfate) and 12 cations (ammonium, monomethylammonium, ethylammonium, potassium, sodium, barium, strontium, magnesium, manganese, calcium, zinc, lead) as the target analytes. These ions are known to be present in postblast residues from inorganic IEDs constructed from ammonium nitrate/fuel oil mixtures, black powder, and chlorate/perchlorate/sugar mixtures. For the analysis of cations, a blue LED (470 nm) was used in conjunction with the highly absorbing cationic dye, chrysoidine (absorption maximum at 453 nm). A nonaqueous background electrolyte comprising 10 mM chrysoidine in methanol was found to give greatly improved baseline stability in comparison to aqueous electrolytes due to the increased solubility of chrysoidine and its decreased adsorption onto the capillary wall. Glacial acetic acid (0.7% v/v) was added to ensure chrysoidine was protonated and to enhance separation selectivity by means of complexation with transition metal ions. The 12 target cations were separated in less than 9.5 min with detection limits of 0.11-2.30 mg/L (calculated at a signal-to-noise ratio of 3). The anions separation system utilized a UV LED (370 nm) in conjunction with an aqueous chromate electrolyte (absorption maximum at 371 nm) consisting of 10 mM chromium(VI) oxide and 10 mM sodium chromate, buffered with 40 mM tris

  15. Spatial Mixture Modelling for Unobserved Point Processes: Examples in Immunofluorescence Histology.

    Science.gov (United States)

    Ji, Chunlin; Merl, Daniel; Kepler, Thomas B; West, Mike

    2009-12-04

    We discuss Bayesian modelling and computational methods in analysis of indirectly observed spatial point processes. The context involves noisy measurements on an underlying point process that provide indirect and noisy data on locations of point outcomes. We are interested in problems in which the spatial intensity function may be highly heterogenous, and so is modelled via flexible nonparametric Bayesian mixture models. Analysis aims to estimate the underlying intensity function and the abundance of realized but unobserved points. Our motivating applications involve immunological studies of multiple fluorescent intensity images in sections of lymphatic tissue where the point processes represent geographical configurations of cells. We are interested in estimating intensity functions and cell abundance for each of a series of such data sets to facilitate comparisons of outcomes at different times and with respect to differing experimental conditions. The analysis is heavily computational, utilizing recently introduced MCMC approaches for spatial point process mixtures and extending them to the broader new context here of unobserved outcomes. Further, our example applications are problems in which the individual objects of interest are not simply points, but rather small groups of pixels; this implies a need to work at an aggregate pixel region level and we develop the resulting novel methodology for this. Two examples with with immunofluorescence histology data demonstrate the models and computational methodology.

  16. Accuracy of the Enhanced Liver Fibrosis Test vs FibroTest, Elastography, and Indirect Markers in Detection of Advanced Fibrosis in Patients With Alcoholic Liver Disease.

    Science.gov (United States)

    Thiele, Maja; Madsen, Bjørn Stæhr; Hansen, Janne Fuglsang; Detlefsen, Sönke; Antonsen, Steen; Krag, Aleksander

    2018-04-01

    Alcohol is the leading cause of cirrhosis and liver-related mortality, but we lack serum markers to detect compensated disease. We compared the accuracy of the Enhanced Liver Fibrosis test (ELF), the FibroTest, liver stiffness measurements (made by transient elastography and 2-dimensional shear-wave elastography), and 6 indirect marker tests in detection of advanced liver fibrosis (Kleiner stage ≥F3). We performed a prospective study of 10 liver fibrosis markers (patented and not), all performed on the same day. Patients were recruited from primary centers (municipal alcohol rehabilitation, n = 128; 6% with advanced fibrosis) and secondary health care centers (hospital outpatient clinics, n = 161; 36% with advanced fibrosis) in the Region of Southern Denmark from 2013 through 2016. Biopsy-verified fibrosis stage was used as the reference standard. The primary aim was to validate ELF in detection of advanced fibrosis in patients with alcoholic liver disease recruited from primary and secondary health care centers, using the literature-based cutoff value of 10.5. Secondary aims were to assess the diagnostic accuracy of ELF for significant fibrosis and cirrhosis and to determine whether combinations of fibrosis markers increase diagnostic yield. The ELF identified patients with advanced liver fibrosis with an area under the receiver operating characteristic curve (AUROC) of 0.92 (95% confidence interval 0.89-0.96); findings did not differ significantly between patients from primary vs secondary care (P = .917). ELF more accurately identified patients with advanced liver fibrosis than indirect marker tests, but ELF and FibroTest had comparable diagnostic accuracies (AUROC of FibroTest, 0.90) (P = .209 for comparison with ELF). Results from the ELF and FibroTest did not differ significantly from those of liver stiffness measurement in intention-to-diagnose analyses (AUROC for transient elastography, 0.90), but did differ in the per-protocol analysis (AUROC for

  17. Quantitative Immunofluorescence Analysis of Nucleolus-Associated Chromatin.

    Science.gov (United States)

    Dillinger, Stefan; Németh, Attila

    2016-01-01

    The nuclear distribution of eu- and heterochromatin is nonrandom, heterogeneous, and dynamic, which is mirrored by specific spatiotemporal arrangements of histone posttranslational modifications (PTMs). Here we describe a semiautomated method for the analysis of histone PTM localization patterns within the mammalian nucleus using confocal laser scanning microscope images of fixed, immunofluorescence stained cells as data source. The ImageJ-based process includes the segmentation of the nucleus, furthermore measurements of total fluorescence intensities, the heterogeneity of the staining, and the frequency of the brightest pixels in the region of interest (ROI). In the presented image analysis pipeline, the perinucleolar chromatin is selected as primary ROI, and the nuclear periphery as secondary ROI.

  18. The Interobserver Agreement between Residents and Experienced Radiologists for Detecting Pulmonary Embolism and DVT with Using CT Pulmonary Angiography and Indirect CT Venography

    International Nuclear Information System (INIS)

    Yavas, Ulas Savas; Calisir, Cuneyt; Ozkan; Ibrahim Ragip

    2008-01-01

    We wanted to prospectively evaluate the interobserver agreement between radiology residents and expert radiologists for interpreting CT images for making the diagnosis of pulmonary embolism (PE). We assessed 112 consecutive patients, from April 2007 to August 2007, who were referred for combined CT pulmonary angiography and indirect CT venography for clinically suspected acute PE. CT scanning was performed with a 64x0.5 collimation multi-detector CT scanner. The CT studies were initially interpreted by the radiology residents alone and then the CT images were subsequently interpreted by a consensus of the resident plus an experienced general radiologist and an experienced chest radiologist. Two of the 112 CTs were unable to be interpreted (1.7%). Pulmonary artery clots were seen on 36 of the thoracic CT angiographies (32%). The interobserver agreement between the radiology residents and the consensus interpretation was good (a kappa index of 0.73). All of the disagreements (15 cases) were instances of overcall by the resident on the initial interpretation. Deep venous thrombosis was detected in 72% (26 of 36) of the patients who had PE seen on thoracic CT. The initial and consensus interpretations of the CT venography images disagreed for two cases (kappa statistic: 0.96). It does not seem adequate to base the final long-term treatment of PE on only the resident's reading, as false positives occurred in 13% of such cases. Timely interpretation of the CT pulmonary angiography and CT venography images should be performed by experienced radiologists for the patients with suspected PE

  19. An investigation of signal performance enhancements achieved through innovative pixel design across several generations of indirect detection, active matrix, flat-panel arrays

    International Nuclear Information System (INIS)

    Antonuk, Larry E.; Zhao Qihua; El-Mohri, Youcef; Du Hong; Wang Yi; Street, Robert A.; Ho, Jackson; Weisfield, Richard; Yao, William

    2009-01-01

    Active matrix flat-panel imager (AMFPI) technology is being employed for an increasing variety of imaging applications. An important element in the adoption of this technology has been significant ongoing improvements in optical signal collection achieved through innovations in indirect detection array pixel design. Such improvements have a particularly beneficial effect on performance in applications involving low exposures and/or high spatial frequencies, where detective quantum efficiency is strongly reduced due to the relatively high level of additive electronic noise compared to signal levels of AMFPI devices. In this article, an examination of various signal properties, as determined through measurements and calculations related to novel array designs, is reported in the context of the evolution of AMFPI pixel design. For these studies, dark, optical, and radiation signal measurements were performed on prototype imagers incorporating a variety of increasingly sophisticated array designs, with pixel pitches ranging from 75 to 127 μm. For each design, detailed measurements of fundamental pixel-level properties conducted under radiographic and fluoroscopic operating conditions are reported and the results are compared. A series of 127 μm pitch arrays employing discrete photodiodes culminated in a novel design providing an optical fill factor of ∼80% (thereby assuring improved x-ray sensitivity), and demonstrating low dark current, very low charge trapping and charge release, and a large range of linear signal response. In two of the designs having 75 and 90 μm pitches, a novel continuous photodiode structure was found to provide fill factors that approach the theoretical maximum of 100%. Both sets of novel designs achieved large fill factors by employing architectures in which some, or all of the photodiode structure was elevated above the plane of the pixel addressing transistor. Generally, enhancement of the fill factor in either discrete or continuous

  20. Routinely used immunoassays do not detect circulating anti-GBM antibodies against native NC1 hexamer and EA epitope of the α3 chain of type IV collagen.

    Science.gov (United States)

    Clavarino, Giovanna; Gauthier, Arnaud; Hellmark, Thomas; Carron, Pierre-Louis; Giovannini, Diane; Colliard, Sophie; Dragon-Durey, Marie-Agnès; Segelmark, Mårten; Cesbron, Jean-Yves; Dumestre-Pérard, Chantal

    2018-04-12

    Detection of circulating anti-GBM antibodies has a key role for the diagnosis of Goodpasture syndrome but immunoassays using purified or recombinant alpha3(IV)NC1 as antigen do not recognize all anti-GBM antibodies. We show that anti-GBM antibodies directed against epitopes in their native conformation or cryptic epitopes are detected by indirect immunofluorescence. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. Nail unit in collagen vascular diseases: A clinical, histopathological and direct immunofluorescence study

    Directory of Open Access Journals (Sweden)

    Nabil P

    2006-01-01

    Full Text Available Background: Abnormalities of the nail unit are common in patients with connective tissue diseases. Clinical examination of the nail unit, coupled with biopsy of proximal nail fold offers an additional advantage in the diagnosis. Purpose: Our aim was to record clinical changes of the nail unit in connective tissue diseases and to study the histopathological (both H and E and periodic acid Schiff and direct immunofluorescence (DIF findings of nail-fold biopsy. Materials and Methods: Thirty-eight confirmed cases connective tissue diseases attending skin OPD were enrolled in the study. After detailed clinical examination of the nail unit, a crescentric biopsy was taken from the proximal nail fold (PNF. Histopathological and DIF studies were was carried out. Findings: Nail changes could be demonstrated in 65% connective tissue diseases. Specific histopathological (H and E and immunofluorescence findings were also encountered in many patients. Conclusion: Clinical examination of the nail unit offers additional clue in the diagnosis of connective tissue diseases. Though DIF of PNF biopsy is useful in the diagnosis, it is not an ideal site for H and E study, as the yield is very low. Limitations: Lack of adequate comparison group and non-utilization of capillary microscopy for the detection of nail fold capillary abnormalities.

  2. Application of a sepharose bead immunofluorescence assay and a solid-phase radioimmunoassay to the bovine leukemia virus system

    International Nuclear Information System (INIS)

    Fiebach, H.; Uckert, W.; Micheel, B.

    1982-01-01

    Several fluorescence assays with bovine leukemia virus (BLV) conjugated to activated Sepharose 4B were used for the detection of BLV and anti-BLV antibodies. These tests were compared with a solid-phase radioimmunoassay and found to be in the same sensitivity range. Sepharose bead immunofluorescence assay and solid-phase radioimmunoassay can be applied to the diagnosis of BLV infection in cattle. (author)

  3. Application of a sepharose bead immunofluorescence assay and a solid-phase radioimmunoassay to the bovine leukemia virus system

    Energy Technology Data Exchange (ETDEWEB)

    Fiebach, H.; Uckert, W.; Micheel, B. (Akademie der Wissenschaften der DDR, Berlin. Zentralinstitut fuer Krebsforschung)

    Several fluorescence assays with bovine leukemia virus (BLV) conjugated to activated Sepharose 4B were used for the detection of BLV and anti-BLV antibodies. These tests were compared with a solid-phase radioimmunoassay and found to be in the same sensitivity range. Sepharose bead immunofluorescence assay and solid-phase radioimmunoassay can be applied to the diagnosis of BLV infection in cattle.

  4. Avaliação sorológica para Toxoplasma gondii pela imunofluorescência indireta e detecção do vírus da imunodeficiência felina pela nested PCR em felinos selvagens Serological evaluation for Toxoplasma gondii by indirect immunofluorescence and detection of feline immunodeficiency virus by nested PCR in wild felines

    Directory of Open Access Journals (Sweden)

    A.V. Rivetti Jr.

    2008-10-01

    Full Text Available Nineteen sera and blood samples from wild feline kept in captivity were tested for Toxoplasma gondii antibody and presence of feline immunodeficiency virus (FIV DNA, respectively. Eighteen (94.7% of the them were seropositive for toxoplasma. However, the only negative animal, a Leopardus pardalis, was the only FIV positve. These results suggest that the infection by FIV may have compromised its immune system and interfered with antibody production for toxoplasma.

  5. IFDOTMETER: A New Software Application for Automated Immunofluorescence Analysis.

    Science.gov (United States)

    Rodríguez-Arribas, Mario; Pizarro-Estrella, Elisa; Gómez-Sánchez, Rubén; Yakhine-Diop, S M S; Gragera-Hidalgo, Antonio; Cristo, Alejandro; Bravo-San Pedro, Jose M; González-Polo, Rosa A; Fuentes, José M

    2016-04-01

    Most laboratories interested in autophagy use different imaging software for managing and analyzing heterogeneous parameters in immunofluorescence experiments (e.g., LC3-puncta quantification and determination of the number and size of lysosomes). One solution would be software that works on a user's laptop or workstation that can access all image settings and provide quick and easy-to-use analysis of data. Thus, we have designed and implemented an application called IFDOTMETER, which can run on all major operating systems because it has been programmed using JAVA (Sun Microsystems). Briefly, IFDOTMETER software has been created to quantify a variety of biological hallmarks, including mitochondrial morphology and nuclear condensation. The program interface is intuitive and user-friendly, making it useful for users not familiar with computer handling. By setting previously defined parameters, the software can automatically analyze a large number of images without the supervision of the researcher. Once analysis is complete, the results are stored in a spreadsheet. Using software for high-throughput cell image analysis offers researchers the possibility of performing comprehensive and precise analysis of a high number of images in an automated manner, making this routine task easier. © 2015 Society for Laboratory Automation and Screening.

  6. In Situ Immunofluorescent Staining of Autophagy in Muscle Stem Cells

    KAUST Repository

    Castagnetti, Francesco

    2017-06-13

    Increasing evidence points to autophagy as a crucial regulatory process to preserve tissue homeostasis. It is known that autophagy is involved in skeletal muscle development and regeneration, and the autophagic process has been described in several muscular pathologies and agerelated muscle disorders. A recently described block of the autophagic process that correlates with the functional exhaustion of satellite cells during muscle repair supports the notion that active autophagy is coupled with productive muscle regeneration. These data uncover the crucial role of autophagy in satellite cell activation during muscle regeneration in both normal and pathological conditions, such as muscular dystrophies. Here, we provide a protocol to monitor the autophagic process in the adult Muscle Stem Cell (MuSC) compartment during muscle regenerative conditions. This protocol describes the setup methodology to perform in situ immunofluorescence imaging of LC3, an autophagy marker, and MyoD, a myogenic lineage marker, in muscle tissue sections from control and injured mice. The methodology reported allows for monitoring the autophagic process in one specific cell compartment, the MuSC compartment, which plays a central role in orchestrating muscle regeneration.

  7. Systemic lupus erythematosus and the Crithidia luciliae immunofluorescent test

    International Nuclear Information System (INIS)

    Whitehouse, I.J.; Fehr, K.; Wagenhaeuser, F.J.

    1983-01-01

    A comparative study of the Crithidia luciliae immunofluorescence (CL-IF) assay and an adapted Farr radioimmunoassay (RIA), for the measurement of antibodies to native deoxyribonucleic acid, was performed using forty-two sera from patients with systematic lupus erythematosus (SLE) and another forty-two from patients with rheumatoid arthritis. Both assays were specific for SLE. The CL-IF assay was statistically significantly more sensitive than the adapted RIA assay. This significant difference was due to greater sensitivity of the CL-IF assay in the cases of sera from patients with SLE of slight activity. Additional advantages of the CL-IF assay were its use to classify the immunoglobulin types of the antibodies (most commonly IgG or IgM) and to measure complement-fixing antibodies to native deoxyribonucleic acid; it affords a simple method of selecting and following SLE patients at risk of developing severe renal disease. These advantages plus the simplicity and inexpensiveness of the CL-IF assay make it a useful tool, especially for use in small laboratories, for the study of antibodies to native deoxyribonucleic acid in patients with SLE. (orig.) [de

  8. Indirection and computer security.

    Energy Technology Data Exchange (ETDEWEB)

    Berg, Michael J.

    2011-09-01

    The discipline of computer science is built on indirection. David Wheeler famously said, 'All problems in computer science can be solved by another layer of indirection. But that usually will create another problem'. We propose that every computer security vulnerability is yet another problem created by the indirections in system designs and that focusing on the indirections involved is a better way to design, evaluate, and compare security solutions. We are not proposing that indirection be avoided when solving problems, but that understanding the relationships between indirections and vulnerabilities is key to securing computer systems. Using this perspective, we analyze common vulnerabilities that plague our computer systems, consider the effectiveness of currently available security solutions, and propose several new security solutions.

  9. Zirconia coated stir bar sorptive extraction combined with large volume sample stacking capillary electrophoresis-indirect ultraviolet detection for the determination of chemical warfare agent degradation products in water samples.

    Science.gov (United States)

    Li, Pingjing; Hu, Bin; Li, Xiaoyong

    2012-07-20

    In this study, a sensitive, selective and reliable analytical method by combining zirconia (ZrO₂) coated stir bar sorptive extraction (SBSE) with large volume sample stacking capillary electrophoresis-indirect ultraviolet (LVSS-CE/indirect UV) was developed for the direct analysis of chemical warfare agent degradation products of alkyl alkylphosphonic acids (AAPAs) (including ethyl methylphosphonic acid (EMPA) and pinacolyl methylphosphonate (PMPA)) and methylphosphonic acid (MPA) in environmental waters. ZrO₂ coated stir bar was prepared by adhering nanometer-sized ZrO₂ particles onto the surface of stir bar with commercial PDMS sol as adhesion agent. Due to the high affinity of ZrO₂ to the electronegative phosphonate group, ZrO₂ coated stir bars could selectively extract the strongly polar AAPAs and MPA. After systematically optimizing the extraction conditions of ZrO₂-SBSE, the analytical performance of ZrO₂-SBSE-CE/indirect UV and ZrO₂-SBSE-LVSS-CE/indirect UV was assessed. The limits of detection (LODs, at a signal-to-noise ratio of 3) obtained by ZrO₂-SBSE-CE/indirect UV were 13.4-15.9 μg/L for PMPA, EMPA and MPA. The relative standard deviations (RSDs, n=7, c=200 μg/L) of the corrected peak area for the target analytes were in the range of 6.4-8.8%. Enhancement factors (EFs) in terms of LODs were found to be from 112- to 145-fold. By combining ZrO₂ coating SBSE with LVSS as a dual preconcentration strategy, the EFs were magnified up to 1583-fold, and the LODs of ZrO₂-SBSE-LVSS-CE/indirect UV were 1.4, 1.2 and 3.1 μg/L for PMPA, EMPA, and MPA, respectively. The RSDs (n=7, c=20 μg/L) were found to be in the range of 9.0-11.8%. The developed ZrO₂-SBSE-LVSS-CE/indirect UV method has been successfully applied to the analysis of PMPA, EMPA, and MPA in different environmental water samples, and the recoveries for the spiked water samples were found to be in the range of 93.8-105.3%. Copyright © 2012 Elsevier B.V. All rights reserved.

  10. Detection of viable and viable nonculturable Vibrio cholerae O1 through cultures and immunofluorescence in the Tucumán rivers, Argentina Detecção de Vibrio cholerae O1 viável e viável não cultivável, através de técnicas de cultivo e imunofluorescência nos rios de Tucumán, Argentina

    Directory of Open Access Journals (Sweden)

    Olga Aulet

    2007-08-01

    Full Text Available Vibrio cholerae has been sporadically isolated from rivers in Tucumán, Argentina, since the outbreak in 1991. The aim of this study was to determine the environmental reservoir of the bacterium in these rivers, assessing the presence of Vibrio cholerae non-O1 and O1 (the latter both in its viable culturable and non culturable state and its relationship to environmental physicochemical variables. 18 water samplings were collected in the Salí River (in Canal Norte and Banda and the Lules River between 2003 and 2005. Physical-chemical measurements (pH, water temperature, electrical conductivity and dissolved oxygen were examined. Vibrio cholerae was investigated with conventional culture methods and with Direct Immunofluorescence (DFA-VNC in order to detect viable non culturable organisms. All isolated microorganisms corresponded to Vibrio cholerae non-O1 and non-O139 (Lules 26%, Canal Norte 33% and Banda 41%. The majority was found during spring and summer and correlated with temperature and pH. Non culturable Vibrio cholerae O1 was detected year round in 38 of the 54 water samples analyzed. Application of the Pearson correlation coefficient revealed that there was no relationship between positive immunofluorescence results and environmental physicochemical parameters. Genes coding for somatic antigen O1 were confirmed in all DFA-VNC-positive samples, whereas the virulence-associated ctxA and tcpA genes were confirmed in 24 samples.Vibrio cholerae tem sido isolado esporadicamente nos rios da Província de Tucumán, Argentina, desde outubro de 1991. O objetivo deste estudo foi localizar os reservatórios nestes rios, identificar a presença de Vibrio cholerae O1 (em estado cultivável e não cultivável e relacionar a presença desta bactéria com as variações físico-químicos da água. Foram coletadas dezoito amostras de água do rio Salí (nas localidades de Canal Norte e Banda e do rio Lules, entre 2003 e 2005. Estas foram submetidas a an

  11. Immunofluorescence pattern of antinuclear antibody and its association with autoantibody profile in systemic lupus erythematosus

    Directory of Open Access Journals (Sweden)

    Sadia Sharmin

    2016-08-01

    Full Text Available Background: Antinuclear antibody (ANA is useful in the diagnosis of systemic lupus erythematosus (SLE. Association of specific autoantibodies with the immunofluorescence pattern of ANA in SLE as noted in Western literature has been taken as reference in all over the world. However, in Bangladesh such research work or data correlating the autoantibodies and their ANA patterns is inadequate. Objective: To identify an association between immunofluorescence patterns of antinuclear antibody on HEp-2 cell and more specific antinuclear reactivities (e.g. anti-dsDNA and anti-extractable nuclear antigen in the serum samples of SLE patients.Methods: Serum samples of 37 SLE patients who were diagnosed by ARA (American Rheumatism Association classification criteria and laboratory tests, attending at lupus clinic of Bangabandhu Sheikh Mujib Medical University (BSMMU during the study period of six months were subjected for ANA testing by Indirect Imrnunofluorescence (IIF on HEp-2 cell, anti-dsDNA by ELISA and anti- extractable nuclear antigen (anti-ENA by Dot Immunoblot. Dot blot strips were tested for anti-Sm, anti-RNP, anti-SSA/Ro, and anti-SSB/La. Results: Out of 37 SLE patients 32 (86.5% cases were ANA positive by IIF on HEp-2 cell. ANA positive sera exhibited three fluorescence patterns such as speckled (43.7%, peripheral (34.3% and homogenous pattern (21.8%. Peripheral pattern (100% was strongly associated with anti-dsDNA (p<0.05 and homogenous pattern (85.7% was also predominantly associated with anti-dsDNA (p<0.05. Speckled pattern (85.6% was significantly associated with anti-ENA (p<0.05. Anti-dsDNA was positive in 75% of SLE cases and majority (45.8% of which showed peripheral pattern whereas anti-ENA was positive in 48.6% cases and majority (70.5% of which showed speckled pattern. The most commonly identified antinuclear autoreactivity was directed towards anti-RNP (22.2% then anti-Sm (16.6%, anti-SSA (16.6% and anti-SSB (11.1 %. Multiple anti

  12. Development of an indirect ELISA to detect Corynebacterium pseudotuberculosis specific antibodies in sheep employing T1 strain culture supernatant as antigen

    Directory of Open Access Journals (Sweden)

    Miriam F. Rebouças

    2013-11-01

    Full Text Available Corynebacterium pseudotuberculosis is the etiologic agent of caseous lymphadenitis (CLA, a chronic disease that affects goats and sheep, characterized by granuloma formation in subcutaneous and internal lymph nodes. CLA causes significant economic losses to commercial goat herds. In this study, we aimed to test secreted antigens secreted from T1 strain bacteria grown in brain heart infusion (BHI broth in an indirect ELISA system to determine the presence of specific immunoglobulins against C. pseudotuberculosis. We analyzed the BHI antigen electrophoretic profile and the recognition pattern by infected sheep sera samples. The ELISA results were compared with multiplex PCR assay and IFN-gamma production. The ELISA was able to discriminate between negative and positive animals, with a sensitivity of 89% and a specificity of 99%, using microbiological isolation as gold standard. When this assay was compared with multiplex PCR and specific IFN-gamma quantification, six discrepant results were found among thirty-two samples. We concluded that the ELISA using antigens secreted from C. pseudotuberculosis T1 strain growth in BHI broth culture can be used for the serodiagnosis of CLA in sheep.

  13. Diagnostic performance of an indirect enzyme-linked immunosorbent assay (ELISA) to detect bovine leukemia virus antibodies in bulk-tank milk samples

    Science.gov (United States)

    Nekouei, Omid; Durocher, Jean; Keefe, Greg

    2016-01-01

    This study assessed the diagnostic performance of a commercial ELISA for detecting bovine leukemia virus antibodies in bulk-tank milk samples from eastern Canada. Sensitivity and specificity of the test were estimated at 97.2% and 100%, respectively. The test was recommended as a cost-efficient tool for large-scale screening programs. PMID:27429469

  14. Sensitivity of direct immunofluorescence in oral diseases. Study of 125 cases.

    Science.gov (United States)

    Sano, Susana Mariela; Quarracino, María Cecilia; Aguas, Silvia Cristina; González, Ernestina Jesús; Harada, Laura; Krupitzki, Hugo; Mordoh, Ana

    2008-05-01

    Direct immunofluorescence (DIF) is widely used for the diagnosis of bullous diseases and other autoimmune pathologies such as oral lichen planus. There is no evidence in the literature on how the following variants influence the detection rate of DIF: intraoral site chosen for the biopsy, perilesional locus or distant site from the clinical lesion, number of biopsies and instrument used. to determine if the following variants influenced the sensitivity (detection rate): intraoral site chosen for the biopsy, perilesional or distant site from the clinical lesion, number of biopsies and instrument used (punch or scalpel). A retrospective study was done at the Cátedra de Patología y Clínica Bucodental II at the Facultad de Odontología, Universidad de Buenos Aires; 136 clinical medical histories were revised for the period March 2000 - March 2005 corresponding to patients with clinical diagnosis of OLP and bullous diseases (vulgar pemphigus, bullous pemphigoid and cicatricial pemphigoid). DIF detection rate was 65.8% in patients with OLP, 66.7% in cicatricial pemphigoid patients, in bullous pemphigoid 55.6%, in pemphigus vulgaris 100%, and in those cases in which certain diagnosis could not be obtained, the DIF positivity rate was 45.5% (Pearson chi(2) (4)= 21.5398 Pr= 0.000). There was no statistically significant difference between the different sites of biopsy (Fisher exact test: 0.825). DIF detection rate in perilesional biopsies was 66.1% and in those distant from the site of clinical lesion was 64.7% (Pearson chi(2) v1)= 0.0073 Pr= 0.932. When the number of biopsies were incremented, DIF detection rate also incremented (Pearson chi(2) = 8.7247 Pr= 0.003). The biopsies taken with punch had a higher detection rate than those taken with scalpel (39.1% versus 71.7%) (Pearson chi(2) = 49.0522 Pr= 0.000). While not statistically significant, the tendency outlined in this study indicates there are intraoral regions in which the detection rate of the DIF technique is

  15. Predicting Outcome and Therapy Response in mCRC Patients Using an Indirect Method for CTCs Detection by a Multigene Expression Panel: A Multicentric Prospective Validation Study

    Directory of Open Access Journals (Sweden)

    Yolanda Vidal Insua

    2017-06-01

    Full Text Available Colorectal cancer (CRC is one of the major causes of cancer-related deaths. Early detection of tumor relapse is crucial for determining the most appropriate therapeutic management. In clinical practice, computed tomography (CT is routinely used, but small tumor changes are difficult to visualize, and reliable blood-based prognostic and monitoring biomarkers are urgently needed. The aim of this study was to prospectively validate a gene expression panel (composed of GAPDH, VIL1, CLU, TIMP1, TLN1, LOXL3 and ZEB2 for detecting circulating tumor cells (CTCs as prognostic and predictive tool in blood samples from 94 metastatic CRC (mCRC patients. Patients with higher gene panel expression before treatment had a reduced progression-free survival (PFS and overall-survival (OS rates compared with patients with low expression (p = 0.003 and p ≤ 0.001, respectively. Patients with increased expression of CTCs markers during treatment presented PFS and OS times of 8.95 and 11.74 months, respectively, compared with 14.41 and 24.7 for patients presenting decreased expression (PFS; p = 0.020; OS; p ≤ 0.001. Patients classified as non-responders by CTCs with treatment, but classified as responders by CT scan, showed significantly shorter survival times (PFS: 8.53 vs. 11.70; OS: 10.37 vs. 24.13; months. In conclusion, our CTCs detection panel demonstrated efficacy for early treatment response assessment in mCRC patients, and with increased reliability compared to CT scan.

  16. Validation of commercially available sphingosine kinase 2 antibodies for use in immunoblotting, immunoprecipitation and immunofluorescence [version 2; referees: 2 approved

    Directory of Open Access Journals (Sweden)

    Heidi A. Neubauer

    2017-03-01

    Full Text Available Sphingosine kinase 2 (SK2 is a ubiquitously expressed lipid kinase that has important, albeit complex and poorly understood, roles in regulating cell survival and cell death. In addition to being able to promote cell cycle arrest and apoptosis under certain conditions, it has recently been shown that SK2 can promote neoplastic transformation and tumorigenesis in vivo. Therefore, well validated and reliable tools are required to study and better understand the true functions of SK2. Here, we compare two commercially available SK2 antibodies: a rabbit polyclonal antibody from Proteintech that recognizes amino acids 266-618 of human SK2a, and a rabbit polyclonal antibody from ECM Biosciences that recognizes amino acids 36-52 of human SK2a. We examine the performance of these antibodies for use in immunoblotting, immunoprecipitation and immunofluorescence staining of endogenous SK2, using human HEK293 and HeLa cell lines, as well as mouse embryonic fibroblasts (MEFs. Furthermore, we assess the specificity of these antibodies to the target protein through the use of siRNA-mediated SK2 knockdown and SK2 knockout (Sphk2-/- MEFs. Our results demonstrate that the Proteintech anti-SK2 antibody reproducibly displayed superior sensitivity and selectivity towards SK2 in immunoblot analyses, while the ECM Biosciences anti-SK2 antibody was reproducibly superior for SK2 immunoprecipitation and detection by immunofluorescence staining. Notably, both antibodies produced non-specific bands and staining in the MEFs, which was not observed with the human cell lines. Therefore, we conclude that the Proteintech SK2 antibody is a valuable reagent for use in immunoblot analyses, and the ECM Biosciences SK2 antibody is a useful tool for SK2 immunoprecipitation and immunofluorescence staining, at least in the human cell lines employed in this study.

  17. Detection of anti-Toxoplasma gondii antibodies in experimentally and naturally infected non-human primates by Indirect Fluorescence Assay (IFA and indirect ELISA Detecção de anticorpos anti-Toxoplasma gondii por meio das técnicas de Imunofluorescência Indireta e ELISA Indireto em primatas experimentalmente e naturalmente infectados

    Directory of Open Access Journals (Sweden)

    Andréa Bouer

    2010-03-01

    Full Text Available The Indirect Fluorescence Assay (IFA and the indirect ELISA were comparatively used to detect IgG and IgM antibodies for Toxoplasma gondii in experimentally and naturally infected primates. In the experimentally infected group, antibodies of diagnostic value were detected at day 9 post-infection (PI with the IFA (IgG and IgM and with IgG-ELISA. IgM-ELISA detected antibodies for T. gondii starting at day 3 PI until the end of the experiment (102 days PI. Of the 209 naturally infected sera tested, from many zoos of State of Sao Paulo, 64.59 and 67.94% were positive in the IgG-IFA test and IgG-ELISA respectively. IgM-ELISA test detected seropositivity in 52.63% of the sera although IgM-IFA test detected it in only in 0.96% of the samples. The differential toxoplasmosis diagnosis was accomplished with Neospora caninum by IFA, observing 61 (29.2% seropositive animals for this parasite and 149 (70.8% negative. Sixty animals were positive for both T. gondii and N. caninum. Pneumonia, splenomegaly, and intestinal ulcers were macroscopically observed. Unremarkable interstitial pneumonia, enteritis, colitis, splenitis, and glomerulitis were microscopically observed. The immunohistochemical stain could not detect the presence of T. gondii in the tissues of the animals infected experimentally.Detectou-se anticorpos das classes IgG e IgM anti-Toxoplasma gondii em primatas experimentalmente e naturalmente infectados, utilizando-se como técnicas comparativas a RIFI e o ELISA-teste. No grupo dos primatas experimentalmente infectados, anticorpos de valor diagnóstico foram detectados a partir do 9º dia de infecção tanto na RIFI (IgG e IgM como no ELISA-IgG. O ELISA IgM detectou anticorpos a partir do 3º dia de infecção até o final do experimento (102 dias pós-infecção. Dos 209 soros dos primatas naturalmente infectados, de diversos zoológicos do Estado de São Paulo, 64,59 e 67,94% mostraram-se positivos na RIFI-IgG e no ELISA-IgG, respectivamente. O

  18. Preparation of anti-Sudan red monoclonal antibody and development of an indirect competitive enzyme-linked immunosorbent assay for detection of Sudan red in chilli jam and chilli oil.

    Science.gov (United States)

    Xu, Jing; Zhang, Yuanyang; Yi, Jian; Meng, Meng; Wan, Yuping; Feng, Caiwei; Wang, Shanliang; Lu, Xiao; Xi, Rimo

    2010-10-01

    Sudan dyes are banned to be used in food additives because of the carcinogenicity of their metabolites. A rapid and sensitive indirect competitive enzyme-linked immunosorbent assay (ELISA) was developed to detect the residues of Sudan dyes. Novel immunogen and coating antigen were synthesized via glutaraldehyde linking. The hapten-bovine serum albumin (BSA) was applied as immunogen and the hapten-ovalbumin (OVA) was served as coating antigen. The monoclonal antibody obtained showed high sensitivity to Sudan I with an IC(50) value of 1.7 μg L(-1) in buffer and was suitable to detect the residues of Sudan red in food products. The specificity of the assay was studied by measuring cross-reactivity of the antibody with the structurally related compounds of Sudan II (red (120%). Chilli jam and chilli oil samples spiked with Sudan dyes were analyzed by the method. The detection limit (LOD) of the ELISA method applied in chilli jam and chilli oil was 9.0 μg L(-1) and 19.6 μg L(-1), respectively. The recovery rates of Sudan-I in chilli oil and chilli jam were in the range of 80%-110% with coefficients of variation <25%. The intra-assay variation and inter-assay variation in buffer were both <9%.

  19. A molecular method for typing Herpes simplex virus isolates as an alternative to immunofluorescence methods

    Directory of Open Access Journals (Sweden)

    Abraham A

    2009-01-01

    Full Text Available Background: Typing of Herpes simplex virus (HSV isolates is required to identify the virus isolated in culture. The methods available for this include antigen detection by immunofluorescence (IF assays and polymerase chain reaction (PCR. This study was undertaken to standardize a molecular method for typing of HSV and compare it with a commercial IF reagent for typing. Objectives: To compare a molecular method for typing HSV isolates with a monoclonal antibody (MAb based IF test. Study design : This cross-sectional study utilized four reference strains and 42 HSV isolates obtained from patients between September 1998 and September 2004. These were subjected to testing using an MAb-based IF test and a PCR that detects the polymerase ( pol gene of HSV isolates. Results: The observed agreement of the MAb IF assay with the pol PCR was 95.7%. Fifty four point eight percent (23/42 of isolates tested by IF typing were found to be HSV-1, 40.5% (17/42 were HSV-2, and two (4.8% were untypable using the MAb IF assay. The two untypable isolates were found to be HSV-2 using the pol PCR. In addition, the cost per PCR test for typing is estimated to be around Rs 1,300 (USD 30, whereas the cost per MAb IF test is about Rs 1,500 (USD 35 including all overheads (reagents, instruments, personnel time, and consumables. Conclusion: The pol PCR is a cheaper and more easily reproducible method for typing HSV isolates as compared to the IF test. It could replace the IF-based method for routine typing of HSV isolates as availability of PCR machines (thermal cyclers is now more widespread than fluorescence microscopes in a country like India.

  20. Indirect field technology for detecting areas object of illegal spills harmful to human health: application of drones, photogrammetry and hydrological models.

    Science.gov (United States)

    Capolupo, Alessandra; Pindozzi, Stefania; Okello, Collins; Boccia, Lorenzo

    2014-12-01

    The accumulation of heavy metals in agricultural soils is a serious environmental problem. The Campania region in southern Italy has higher levels of cancer risk, presumably due to the accumulation of geogenic and anthropogenic soil pollutants, some of which have been incorporated into organic matter. The aim of this study was to introduce and test an innovative, field-applicable methodology to detect heavy metal accumulation using drone-based photogrammetry and microrill network modelling, specifically to generate wetlands wetlands prediction indices normally applied at large catchment scales, such as a large geographic basin. The processing of aerial photos taken using a hexacopter equipped with fifth-generation software for photogrammetry allowed the generation of a digital elevation model (DEM) with a resolution as high as 30 mm. Not only this provided a high potential for the study of micro-rill processes, but it was also useful for testing and comparing the capability of the topographic index (TI) and the clima-topographic index (CTI) to predict heavy metal sedimentation points at scales from 0.1 to 10 ha. Our results indicate that the TI and CTI indices can be used to predict points of heavy metal accumulation for small field catchments.

  1. Pedal indirect lymphangiography

    International Nuclear Information System (INIS)

    Lee, Kil Woo; Hong, Myung Sun; Kim, In Jae

    1994-01-01

    Recently, indirect lymphangiography has been developed as a relatively good and noninvasive imaging modality of the lymphatic system at extremities. But the disadvantage of the indirect lymphangiography is a low contrast ratio between the surrounding tissues and the contrast media in lymphatic vessels, because dimeric nonionic contrast media is water soluble and diluted in the proximal leg lymphatic vessels. We could have relatively better image than previously published images for the leg lymphatic system, when we injected contrast media with adequate high pressure in intradermal space of the interdigital areas at the foot dorsum. So, we would like to report the results. We could study all 9 lymphedemas(primary: 6, secondary: 3) from April 1990 to May 1993 on outpatient base. They were diagnosed as lymphedema clinically and radiologically. Ten ml of dimeric nonionic aget, iotrolan(Isovist 300) was injected into intradermal space with five 30-gauge needles. The injection speed was more than 0.2 ml/min. We have done one side pedal lymphangiogram in 30 minutes. The evaluation of the anterior superficial lymphatics was according to the criteria of the Weissleder. The results were as follows: 1. All lymphatic vessels from foot to inguinal area could be visualized. 2. Two or three inferior inguinal lymph nodes could be visualized about 42%. 3. The most common abnormal finding of the lymphedma was the neovascularization of the lymphatics on indirect pedal lymphangiogram. If we use adequate technique relatively high pressure injection, correct intradermal needle insertion, adequate soft tissue exposure technique indirect lymphangiography is considered to be a safe and noninvasive imaging modality for the evaluation of the lymphedema of lower extremity lymphatics including inferior inguinal lymph nodes

  2. Development of an indirect spectrophotometric method for ...

    African Journals Online (AJOL)

    A simple and rapid indirect spectrophotometric method for determination of ... of the colored product was measured at 405 nm and pH 3 against a reagent blank. ... The limit of detection and quantification were found to be 0.20±0.03 and ...

  3. CONTRIBUTION OF INDIRECT TAXES

    Directory of Open Access Journals (Sweden)

    CHIRCULESCU MARIA FELICIA

    2015-08-01

    Full Text Available The work is based on the fact that at any time and in any society, taxation is regarded as undesirable for all taxpayers. The existence and it's manifestation is justified, because the operation of any company involves costs that must be covered by sufficient resources. Since ancient times, each state has adopted its own tax system, more or less perfected, as the state has experienced a greater or lesser economic and military power At the base of this work stays the fact that tax systems are a key factor influencing the overall efficiency of the economy. They determine the size tendency to save, invest and work, influencing the increase in production and employment, which is essential sights integral economic strategy, making tax reform an important component of economic reform. This paper aims to analyze the indirect taxes and their contribution to the public revenues in Romania, the purpose paper contains an analysis based on statistical series as indirect taxation is where tax harmonization was possible. Through analyzes, the paper aims to provide answers to the problem of the contradiction between the growing need for budgetary revenues, which entails a continuous amplification and diversification of taxation, on the one hand, and the need to stimulate economic development, on the other hand. The harmonization of indirect taxation had been achieved since this touches the free movement of goods and the freedom to supply services, not being able to say the same thing about direct taxation, which is why the European Community Treaty does not specify expressly the alignment of direct taxation, considering that direct taxation is a matter of Internal Policies that, for a country free option.

  4. Detection of antinuclear antibodies by solid-phase immunoassays and immunofluorescence analysis

    DEFF Research Database (Denmark)

    Fenger, Mogens; Wiik, Allan; Høier-Madsen, Mimi

    2004-01-01

    -established rheumatic disorders, the newly developed EIA in which HEp-2 extracts were included had sensitivities and specificities comparable to or in some instances better than the IFA. The assays without HEp-2 extracts included had significantly lower sensitivities and specificities. In the outpatient population, up...... to 51% of patients had positive ANA tests that did not correspond to classic ANA-associated disease. However, in the assays in which the HEp-2 extracts were not included, the false-positive rate was ... and was mostly extracts included had a low sensitivity but a high specificity, particularly in nonselected populations. The choice of test is highly dependent on the clinical setting in which...

  5. Detection of Bim and Puma in mouse hair follicles using immunofluorescence and TUNEL assay double staining

    Czech Academy of Sciences Publication Activity Database

    Veselá, Barbora; Matalová, Eva

    2015-01-01

    Roč. 90, č. 8 (2015), s. 587-593 ISSN 1052-0295 Grant - others:GA ČR(CZ) GAP502/12/1285 Program:GA Institutional support: RVO:67985904 Keywords : apoptosis * Bim * development Subject RIV: ED - Physiology Impact factor: 1.078, year: 2015

  6. Detection of the thraustochytrid protist Ulkenia visurgensis in a hydroid, using immunofluorescence

    Digital Repository Service at National Institute of Oceanography (India)

    Raghukumar, S.

    to November 1986 By treating the samples with antiserum prepared against this organism and conjugated with FITC stain, the protist was regularly found to occur in association with a hydroid Several cells of the organism were observed in the coelenteron...

  7. Studies on direct and indirect electrochemical immunoassays

    OpenAIRE

    Buckley, Eileen

    1989-01-01

    Two approaches to electrochemical immunoassay are reported. The first approach was an indirect method, involving an electroactive, enzyme-catalysed, substrate to product reaction. Conditions were optimised for the amperometric detection of para-aminophenol, the electroactive product of the alkaline phosphatase catalysed hydrolysis of a new substrate, p-aminophenylphosphate, after separation by HPLC. The second approach involved the direct electrochemical detection of an immunoglo...

  8. Immunofluorescent histological studies of the role of fibronectin in the expression of the associative preferences of embryonic tissues.

    Science.gov (United States)

    Armstrong, P B; Armstrong, M T

    1981-08-01

    The identity of the chemical factors controlling the spreading behaviour of sheets of cells was examined in organ culture. When aggregates of two dissimilar tissues are apposed in organ culture, one tissue spreads reproducibly over the surface of the second. The present study employed indirect immunofluorescent localization techniques to evaluate the hypothesis that the spreading behaviour of chick embryonic heart tissue in culture is dominated by the presence or absence of the cell-surface and extracellular matrix protein fibronectin in the surface layers of the aggregates. Specifically, the hypothesis proposes that aggregates that display surface fibronectin earlier after culturing and/or in higher quantities segregate internally to aggregates that are slower to develop a surface layer of fibronectin or in which this layer contains reduced amounts of fibronectin. The hypothesis has been supported for 3 categories of behaviour of chick embryo heart tissue: (1) myocyte aggregates spread over myocyte aggregates containing a 20% admixture of heart fibroblasts, which in turn spread over heart fibroblast aggregates; (2) 5-day embryonic ventricle-tissue fragments maintained in culture for 0.5 days spread over ventricle fragments cultured for 2.5 days; and (3) 2-day embryonic ventricle spreads over 5-day ventricle. In all these situations, the aggregate type that segregates to an internal position displays more fibronectin at its surface than aggregate types that spread to occupy an external position. Evidence is presented that the fibronectin in heart tissue aggregates is elaborated by heart fibroblasts.

  9. GP50 as a promising early diagnostic antigen for Taenia multiceps infection in goats by indirect ELISA.

    Science.gov (United States)

    Huang, Xing; Xu, Jing; Wang, Yu; Guo, Cheng; Chen, Lin; Gu, Xiaobin; Lai, Weimin; Peng, Xuerong; Yang, Guangyou

    2016-12-01

    Coenurosis is caused by coenurus, the metacestode of Taenia multiceps, which mainly parasitizes the brain and spinal cord of cattle, sheep and goats. To date, no widely-approved methods are available to identify early coenurus infection. In this study, we identified a full-length cDNA that encodes GP50 (TmGP50) from the transcriptome of T. multiceps, and then cloned and expressed in E. coli. The native proteins in adult stage and coenurus were located via immunofluorescence assays, while the potential of recombinant TmGP50 protein (rTmGP50) for indirect ELISA-based serodiagnostics was assessed using native goat sera. In addition, we orally infected 20 goats with mature T. multiceps eggs. Praziquantel (10%) was given to 10 of the goats 45 days post-infection (p.i.). Blood samples were collected for 17 weeks p.i. from the 20 goats and anti-rTmGP50 antibodies were evaluated using the indirect ELISA established here. The TmGP50 contains an 897 bp open reading frame, in which signal sequence resides in 1 ~ 48 sites and mature polypeptide consists of 282 amino acid residues. Immunofluorescence staining showed that native TmGP50 was localized to the microthrix and parenchymatous zone of the adult parasite and coenurus, and the coenurus cystic wall. The indirect ELISA based on rTmGP50 exhibited a sensitivity of 95.0% and a specificity of 92.6% when detecting GP50 antibodies in sera of naturally infected goats and sheep. In goats experimentally infected with T. multiceps, anti-TmGP50 antibody was detectable from 2 to 17 weeks p.i. in the control group, while the antibody fell below the cut-off value about 3 weeks after praziquantel treatment. Our results indicate that recombinant TmGP50 is a suitable early diagnostic antigen for coenurus infection in goats.

  10. Indirect detection of heavy supersymmetric dark matter

    International Nuclear Information System (INIS)

    Kamionkowski, M.

    1991-02-01

    If neutralinos reside in the galactic halo they will be captured in the Sun and annihilate therein producing high-energy neutrinos. Present limits on the flux of such neutrinos from underground detectors such as IMB and Kamiokande 2 may be used to rule out certain supersymmetric dark-matter candidates, while in many other supersymmetric models the rates are large enough that if neutralinos do reside in the galactic halo, observation of a neutrino signal may be possible in the near future. 10 refs., 2 figs

  11. Comparison of immunofluorescence investigations and a new anti-DNA-antibody radioimmunoassay for the diagnosis of connective tissue diseases

    International Nuclear Information System (INIS)

    Neumeier, D.; Vogt, W.; Knedel, M.

    1976-01-01

    The procedure of determining the anti-DNA antibody activity is simplified by high-molecular double strand DNA labelled with 125 I. Cases of suspected connective tissue disease should first be examined by immunofluorescence microscopy, since this method can detect a wider spectrum of diseases with similar symptoms. For a differential diagnosis of SLE, the anti-DNA antibody activity is then investigated by a radioimmunoassay. When assessing the antibody activity, the following criteria should be kept in mind: - Findings of less than 10 units/ml serum do not indicate pathological changes, - Higher antibody activities up to 35 units/ml serum may occur in SLE patients but are also possible in other ANA positive diseases with similar symptoms, - Activities over 35 units/ml serum are nearly always a sign of SLE. (orig./GSE) [de

  12. Comparison of immunofluorescence investigations and a new anti-DNA-antibody radioimmunoassay for the diagnosis of connective tissue diseases

    Energy Technology Data Exchange (ETDEWEB)

    Neumeier, D; Vogt, W; Knedel, M

    1976-01-01

    The procedure of determining the anti-DNA antibody activity is simplified by high-molecular double strand DNA labelled with /sup 125/I. Cases of suspected connective tissue disease should first be examined by immunofluorescence microscopy, since this method can detect a wider spectrum of diseases with similar symptoms. For a differential diagnosis of SLE, the anti-DNA antibody activity is then investigated by a radioimmunoassay. When assessing the antibody activity, the following criteria should be kept in mind: - Findings of less than 10 units/ml serum do not indicate pathological changes, - Higher antibody activities up to 35 units/ml serum may occur in SLE patients but are also possible in other ANA positive diseases with similar symptoms, - Activities over 35 units/ml serum are nearly always a sign of SLE.

  13. BLIND TRIALS EVALUATING IN VITRO INFECTIVITY OF CRYPTOSPORIDIUM PARVUM OOCYSTS USING CELL CULTURE IMMUNOFLUORESCENCE

    Science.gov (United States)

    An optimized cell culture-immunofluorescence (IFA) procedure, using the HCT-8 cell line, was evaluated in 'blind' trials to determine the sensitivity and reproducibility for measuring infectivity of flow cytometry prepared inocula of C. parvum oocysts. In separate trials, suspens...

  14. WE-G-204-05: Relative Object Detectability Evaluation of a New High Resolution A-Se Direct Detection System Compared to Indirect Micro-Angiographic Fluoroscopic (MAF) Detectors

    International Nuclear Information System (INIS)

    Russ, M; Nagesh, S Setlur; Ionita, C; Bednarek, D; Rudin, S; Scott, C; Karim, K

    2015-01-01

    Purpose: To evaluate the task specific imaging performance of a new 25µm pixel pitch, 1000µm thick amorphous selenium direct detection system with CMOS readout for typical angiographic exposure parameters using the relative object detectability (ROD) metric. Methods: The ROD metric uses a simulated object function weighted at each spatial frequency by the detectors’ detective quantum efficiency (DQE), which is an intrinsic performance metric. For this study, the simulated objects were aluminum spheres of varying diameter (0.05–0.6mm). The weighted object function is then integrated over the full range of detectable frequencies inherent to each detector, and a ratio is taken of the resulting value for two detectors. The DQE for the 25µm detector was obtained from a simulation of a proposed a-Se detector using an exposure of 200µR for a 50keV x-ray beam. This a-Se detector was compared to two microangiographic fluoroscope (MAF) detectors [the MAF-CCD with pixel size of 35µm and Nyquist frequency of 14.2 cycles/mm and the MAF-CMOS with pixel size of 75µm and Nyquist frequency of 6.6 cycles/mm] and a standard flat-panel detector (FPD with pixel size of 194µm and Nyquist frequency of 2.5cycles/mm). Results: ROD calculations indicated vastly superior performance by the a-Se detector in imaging small aluminum spheres. For the 50µm diameter sphere, the ROD values for the a-Se detector compared to the MAF-CCD, the MAF-CMOS, and the FPD were 7.3, 9.3 and 58, respectively. Detector performance in the low frequency regime was dictated by each detector’s DQE(0) value. Conclusion: The a-Se with CMOS readout is unique and appears to have distinctive advantages of incomparable high resolution, low noise, no readout lag, and expandable design. The a-Se direct detection system will be a powerful imaging tool in angiography, with potential break-through applications in diagnosis and treatment of neuro-vascular disease. Supported by NIH Grant: 2R01EB002873 and an

  15. WE-G-204-05: Relative Object Detectability Evaluation of a New High Resolution A-Se Direct Detection System Compared to Indirect Micro-Angiographic Fluoroscopic (MAF) Detectors

    Energy Technology Data Exchange (ETDEWEB)

    Russ, M; Nagesh, S Setlur; Ionita, C; Bednarek, D; Rudin, S [Toshiba Stroke and Vascular Research Center, University at Buffalo (SUNY), Buffalo, NY (United States); Scott, C; Karim, K [University of Waterloo, Waterloo, ON (Canada)

    2015-06-15

    Purpose: To evaluate the task specific imaging performance of a new 25µm pixel pitch, 1000µm thick amorphous selenium direct detection system with CMOS readout for typical angiographic exposure parameters using the relative object detectability (ROD) metric. Methods: The ROD metric uses a simulated object function weighted at each spatial frequency by the detectors’ detective quantum efficiency (DQE), which is an intrinsic performance metric. For this study, the simulated objects were aluminum spheres of varying diameter (0.05–0.6mm). The weighted object function is then integrated over the full range of detectable frequencies inherent to each detector, and a ratio is taken of the resulting value for two detectors. The DQE for the 25µm detector was obtained from a simulation of a proposed a-Se detector using an exposure of 200µR for a 50keV x-ray beam. This a-Se detector was compared to two microangiographic fluoroscope (MAF) detectors [the MAF-CCD with pixel size of 35µm and Nyquist frequency of 14.2 cycles/mm and the MAF-CMOS with pixel size of 75µm and Nyquist frequency of 6.6 cycles/mm] and a standard flat-panel detector (FPD with pixel size of 194µm and Nyquist frequency of 2.5cycles/mm). Results: ROD calculations indicated vastly superior performance by the a-Se detector in imaging small aluminum spheres. For the 50µm diameter sphere, the ROD values for the a-Se detector compared to the MAF-CCD, the MAF-CMOS, and the FPD were 7.3, 9.3 and 58, respectively. Detector performance in the low frequency regime was dictated by each detector’s DQE(0) value. Conclusion: The a-Se with CMOS readout is unique and appears to have distinctive advantages of incomparable high resolution, low noise, no readout lag, and expandable design. The a-Se direct detection system will be a powerful imaging tool in angiography, with potential break-through applications in diagnosis and treatment of neuro-vascular disease. Supported by NIH Grant: 2R01EB002873 and an

  16. Determination of aminopolycarboxylic acids at ultra-trace levels by means of online coupling ion exchange chromatography and inductively coupled plasma-mass spectrometry with indirect detection via their Pd2+-complexes

    International Nuclear Information System (INIS)

    Nette, David; Seubert, Andreas

    2015-01-01

    Highlights: • 8 important APCA’s analyzed in one run instead of 3 in the previous method. • Pd 2+ extents the methods applicability to 3 and more dentate amino carboxylic acids. • Separation system optimized for the isocratic determination of important APCA’s. • Thermodynamic stability of APCA–Pd 2+ complexes is higher than for Fe 3+ and In 3+ . • Pd 2+ is kinetically much slower than Fe 3+ and In 3+ and makes the method more rugged. - Abstract: A new indirect IC-ICP-MS method for the determination of aminopolycarboxylic acids in water samples is described. It is based on the addition of an excess of Pd(II) to water samples. The analytes are forced into very strong and negatively charged palladium complexes, separated by ion exchange chromatography and detected by their palladium content, utilizing an on-line coupled ICP-MS. This method is suitable to determine the concentration of 8 aminopolycarboxylic acids (nitrilotriacetic acid (NTA), (2-carboxyethyl) iminodiacetic acid (β-ADA), methylglycinediacetic acid (MGDA), 2-hydroxyethyl) ethylenediamine triacetic acid (HEDTA), diethylene triamine pentaacetic acid (DTPA), ethylendiamine tetraacetic acid (EDTA), 1,3-diaminopropane tetraacetic acid (1,3-PDTA) and 1,2-diaminopropane tetraacetic acid (1,2-PDTA) at the ng kg −1 level. The method is faster and easier than the established gas chromatography (GC)-method ISO 16588:2002 [1] and up to two orders of magnitude more sensitive than the ion pair chromatography based method of DIN 38413-8. Analytic performance is superior to ISO 16588:2002 and the comparability is good

  17. Determination of aminopolycarboxylic acids at ultra-trace levels by means of online coupling ion exchange chromatography and inductively coupled plasma-mass spectrometry with indirect detection via their Pd{sup 2+}-complexes

    Energy Technology Data Exchange (ETDEWEB)

    Nette, David; Seubert, Andreas, E-mail: seubert@staff.uni-marburg.de

    2015-07-16

    Highlights: • 8 important APCA’s analyzed in one run instead of 3 in the previous method. • Pd{sup 2+} extents the methods applicability to 3 and more dentate amino carboxylic acids. • Separation system optimized for the isocratic determination of important APCA’s. • Thermodynamic stability of APCA–Pd{sup 2+} complexes is higher than for Fe{sup 3+} and In{sup 3+}. • Pd{sup 2+} is kinetically much slower than Fe{sup 3+} and In{sup 3+} and makes the method more rugged. - Abstract: A new indirect IC-ICP-MS method for the determination of aminopolycarboxylic acids in water samples is described. It is based on the addition of an excess of Pd(II) to water samples. The analytes are forced into very strong and negatively charged palladium complexes, separated by ion exchange chromatography and detected by their palladium content, utilizing an on-line coupled ICP-MS. This method is suitable to determine the concentration of 8 aminopolycarboxylic acids (nitrilotriacetic acid (NTA), (2-carboxyethyl) iminodiacetic acid (β-ADA), methylglycinediacetic acid (MGDA), 2-hydroxyethyl) ethylenediamine triacetic acid (HEDTA), diethylene triamine pentaacetic acid (DTPA), ethylendiamine tetraacetic acid (EDTA), 1,3-diaminopropane tetraacetic acid (1,3-PDTA) and 1,2-diaminopropane tetraacetic acid (1,2-PDTA) at the ng kg{sup −1} level. The method is faster and easier than the established gas chromatography (GC)-method ISO 16588:2002 [1] and up to two orders of magnitude more sensitive than the ion pair chromatography based method of DIN 38413-8. Analytic performance is superior to ISO 16588:2002 and the comparability is good.

  18. Fibrinogen Demonstration in Oral Lichen Planus: An Immunofluorescence Study on Archival Tissues.

    Science.gov (United States)

    Shirol, Pallavi D; Naik, Veena; Kale, Alka

    2015-10-01

    Lichen planus is a premalignant condition with minimal diagnostic aids. This study is an attempt to use paraffin embedded sections of lichen planus with immunofluorescein stain and to evaluate the immunofluorescent sections to establish pattern of fibrinogen deposition. Thirty-five paraffin embedded sections of old and new cases of oral lichen planus (study group) and five normal oral mucosa (control group) were chosen. Two sections of each (H & E) case were taken, one was stained with hematoxylin and eosin and another with fluorescein isothiocynate conjugate (FITC) polyclonal rabbit antibody against fibrinogen. Fluorescent findings were examined with a fluorescent microscope. A high statistical significant correlation was found in respect to fluorescence positivity, intensity of fluorescence and distribution of fluorescence each with p < 0.0001 and fluorescence at blood vessel walls (p = 0.0003). This study suggested that paraffin embedded sections can be successfully used in direct immunofluorescence staining in routine set up where only formalin fixed tissues are received. Paraffin embedded sections can be successfully used in direct immunofluorescence staining when only formalin fixed tissues are received.

  19. A novel assay for detecting antibodies to cytochrome P4502D6, the molecular target of liver kidney microsomal antibody type 1.

    Science.gov (United States)

    Kerkar, N; Ma, Y; Hussain, M; Muratori, L; Targett, C; Williams, R; Bianchi, F B; Mieli-Vergani, G; Vergani, D

    1999-03-04

    Liver Kidney Microsomal type 1 (LKM1) antibody, the diagnostic marker of autoimmune hepatitis type 2, is also found in a proportion of patients with hepatitis C virus infection (HCV). It is detected conventionally by the subjective immunofluorescence technique. Our aim was to establish a simple and objective enzyme-linked immunosorbent assay (ELISA) that measures antibodies to cytochrome P4502D6 (CYP2D6), the target of LKM1. An indirect ELISA using eukaryotically expressed CYP2D6 was designed. Absorbance values obtained against a reference microsomal preparation were subtracted from those obtained against a microsomal preparation over-expressing CYP2D6, thus removing the non-CYP2D6-specific reaction. Sera from 51 LKM1 positive patients (21 autoimmune hepatitis and 30 with HCV infection), 111 LKM1 negative patients with chronic liver disease (including 20 with HCV infection) and 43 healthy controls were tested. Of 51 patients positive by immunofluorescence, 48 were also positive by ELISA while all the 154 LKM1 negative subjects were also negative by ELISA. There was a high degree of association between IFL and ELISA as demonstrated by a kappa reliability value of 0.96. The absorbance values by ELISA correlated with immunofluorescence LKM1 titres both in autoimmune hepatitis (r = 0.74, p < 0.001) and HCV infection (r = 0.67, p < 0.001). The simple, objective ELISA described has the potential to replace the standard immunofluorescence technique.

  20. Direct immunofluorescence and enzyme-linked immunosorbent assays for evaluating chlorinated hydrocarbon degrading bacteria

    Energy Technology Data Exchange (ETDEWEB)

    Brigmon, R.L.; Franck, M.M.; Brey, J.; Fliermans, C.B. [Westinghouse Savannah River, Aiken, SC (United States). Environmental Biotechnology Section; Scott, D.; Lanclos, K. [Medical Coll. of Georgia, Augusta, GA (United States)

    1997-06-01

    Immunological procedures were developed to enumerate chlorinated hydrocarbon degrading bacteria. Polyclonal antibodies (Pabs) were produced by immunizing New Zealand white rabbits against 18 contaminant-degrading bacteria. These included methanotrophic and chlorobenzene (CB) degrading species. An enzyme-linked immunosorbent assay (ELISA) was used to test for specificity and sensitivity of the Pabs. Direct fluorescent antibodies (DFAs) were developed with these Pabs against select methanotrophic bacteria isolated from a trichloroethylene (TCE) contaminated landfill at the Savannah River Site (SRS) and cultures from the American Type Culture Collection (ATCC). Analysis of cross reactivity testing data showed some of the Pabs to be group specific while others were species specific. The threshold of sensitivity for the ELISA is 105 bacteria cells/ml. The DFA can detect as few as one bacterium per ml after concentration. Results from the DFA and ELISA techniques for enumeration of methanotrophic bacteria in groundwater were higher but not significantly different (P < 0.05) compared to indirect microbiological techniques such as MPN. These methods provide useful information on in situ community structure and function for bioremediation applications within 1--4 hours of sampling.

  1. Indirect search for neutralino dark matter with high energy neutrinos

    International Nuclear Information System (INIS)

    Barger, V.; Halzen, Francis; Hooper, Dan; Kao, Chung

    2002-01-01

    We investigate the prospects of indirect searches for supersymmetric neutralino dark matter. Relic neutralinos gravitationally accumulate in the Sun and their annihilations produce high energy neutrinos. Muon neutrinos of this origin can be seen in large detectors such as AMANDA, IceCube, and ANTARES. We evaluate the relic density and the detection rate in several models--the minimal supersymmetric model, minimal supergravity, and supergravity with nonuniversal Higgs boson masses at the grand unification scale. We make realistic estimates for the indirect detection rates including effects of the muon detection threshold, quark hadronization, and solar absorption. We find good prospects for detection of neutralinos with mass above 200 GeV

  2. The Complexity of Indirect Translation

    DEFF Research Database (Denmark)

    Wenjie, L. I.

    2017-01-01

    its complex nature, and thus determined that many facets of ITr remain to be studied. The present article will try to encompass the complexity of ITr by looking into the reasons for translating indirectly, the challenge of finding out mediating texts (MTs), indirectness in both translation...... of which have been translated and interpreted indirectly through major languages like English, will be employed as examples. Hopefully, this study will offer more insights into the nature of translation as a social activity and raise further interests in studying translation as a complex phenomenon....

  3. A simplified indirect bonding technique

    Directory of Open Access Journals (Sweden)

    Radha Katiyar

    2014-01-01

    Full Text Available With the advent of lingual orthodontics, indirect bonding technique has become an integral part of practice. It involves placement of brackets initially on the models and then their transfer to teeth with the help of transfer trays. Problems encountered with current indirect bonding techniques used are (1 the possibility of adhesive flash remaining around the base of the brackets which requires removal (2 longer time required for the adhesive to gain enough bond strength for secure tray removal. The new simplified indirect bonding technique presented here overcomes both these problems.

  4. ELISA indireto para detecção de IgG antivírus da doença de Newcastle em soro de codorna Indirect ELISA for the detection of IgG specific to Newcastle disease virus in quail serum

    Directory of Open Access Journals (Sweden)

    D.D. Oliveira

    2007-10-01

    Full Text Available An indirect ELISA for the detection of japanese quail IgG specific to Newcastle disease virus (NDV was developed. The secondary anti-quail IgG was produced in Balb/c mice, by inoculating Freund's complete adjuvant emulsified japanese quail-IgG extract. The purification of IgG was achieved using the caprilic acid method. The ELISA was compared to the haemagglutination-inhibition (HI test for antibodies to NDV. ELISA cut-off point was established through TG-ROC analysis. Total correlation was observed between the ELISA and the HI, being the ELISA efficient in the identification of positive and negative sera, with high sensitivity and specificity (100%. These results validate the use of the indirect ELISA as an alternative for the detection of NDV-specific IgG in japanese quail sera, with the advantage of high sensitivity and automation.

  5. The logic of indirect speech

    Science.gov (United States)

    Pinker, Steven; Nowak, Martin A.; Lee, James J.

    2008-01-01

    When people speak, they often insinuate their intent indirectly rather than stating it as a bald proposition. Examples include sexual come-ons, veiled threats, polite requests, and concealed bribes. We propose a three-part theory of indirect speech, based on the idea that human communication involves a mixture of cooperation and conflict. First, indirect requests allow for plausible deniability, in which a cooperative listener can accept the request, but an uncooperative one cannot react adversarially to it. This intuition is supported by a game-theoretic model that predicts the costs and benefits to a speaker of direct and indirect requests. Second, language has two functions: to convey information and to negotiate the type of relationship holding between speaker and hearer (in particular, dominance, communality, or reciprocity). The emotional costs of a mismatch in the assumed relationship type can create a need for plausible deniability and, thereby, select for indirectness even when there are no tangible costs. Third, people perceive language as a digital medium, which allows a sentence to generate common knowledge, to propagate a message with high fidelity, and to serve as a reference point in coordination games. This feature makes an indirect request qualitatively different from a direct one even when the speaker and listener can infer each other's intentions with high confidence. PMID:18199841

  6. Comparison between immunomagnetic separation, coupled with immunofluorescence, and the techniques of Faust et al. and of Lutz for the diagnosis of Giardia lamblia cysts in human feces

    Directory of Open Access Journals (Sweden)

    Souza Doris Sobral Marques

    2003-01-01

    Full Text Available In the present study, the performance of Immunomagnetic Separation technique, coupled with Immunofluorescence (IMS-IFA, was compared with the FAUST et al. and Lutz parasitological techniques for the detection of Giardia lamblia cysts in human feces. One hundred and twenty-seven samples were evaluated by the three techniques at the same time showing a rate of cyst detection of 27.5% by IMS-IFA and 15.7% by both Faust et al. and Lutz techniques. Data analysis showed a higher sensitivity of IMS-IFA for the detection of G. lamblia cysts in comparison with the techniques of FAUST et al. and Lutz. The use of this methodology as a routine procedure enables the processing of many samples simultaneously, in order to increase recovery rate of G. lamblia cysts and reduce the time of sample storage.

  7. Development and application of an indirect enzyme-linked immunosorbent assay using recombinant truncated Cap protein for the diagnosis of porcine circovirus-like virus P1.

    Science.gov (United States)

    Wen, Li-bin; Wen, Shi-fu; He, Kong-wang

    2016-01-19

    Porcine circovirus-like virus P1 is a newly discovered virus. To date, there has been no specific serological assay for use in the diagnosis of P1 infection. Because P1 has high homology to porcine circovirus type 2 (PCV2) at the nucleotide level, the C-terminal portion of the capsid protein (amino acids 73-114), a discriminative antigen, was expressed in a prokaryotic expression system. The recombinant product (rctCap), composed of three identical repeated domains, was shown to be strongly immunoreactive to P1-specific serum. This assay was validated by comparison with an indirect immunofluorescence assay (IFA). The diagnostic sensitivity and specificity of the rctCap enzyme-linked immunosorbent assay (ELISA) developed in this study are 93.6% and 98.3%, respectively, compared with the results from IFAs on 450 sera samples from pigs. The indirect ELISA that we developed with rctCap, the recombinant capsid fragment containing the 217-342 nt repeat domain, was sensitive, specific, and suitable for the large-scale detection of P1 infections in swine.

  8. Semi-automated relative quantification of cell culture contamination with mycoplasma by Photoshop-based image analysis on immunofluorescence preparations.

    Science.gov (United States)

    Kumar, Ashok; Yerneni, Lakshmana K

    2009-01-01

    Mycoplasma contamination in cell culture is a serious setback for the cell-culturist. The experiments undertaken using contaminated cell cultures are known to yield unreliable or false results due to various morphological, biochemical and genetic effects. Earlier surveys revealed incidences of mycoplasma contamination in cell cultures to range from 15 to 80%. Out of a vast array of methods for detecting mycoplasma in cell culture, the cytological methods directly demonstrate the contaminating organism present in association with the cultured cells. In this investigation, we report the adoption of a cytological immunofluorescence assay (IFA), in an attempt to obtain a semi-automated relative quantification of contamination by employing the user-friendly Photoshop-based image analysis. The study performed on 77 cell cultures randomly collected from various laboratories revealed mycoplasma contamination in 18 cell cultures simultaneously by IFA and Hoechst DNA fluorochrome staining methods. It was observed that the Photoshop-based image analysis on IFA stained slides was very valuable as a sensitive tool in providing quantitative assessment on the extent of contamination both per se and in comparison to cellularity of cell cultures. The technique could be useful in estimating the efficacy of anti-mycoplasma agents during decontaminating measures.

  9. A quantum dot-immunofluorescent labeling method to investigate the interactions between a crinivirus and its whitefly vector

    Directory of Open Access Journals (Sweden)

    James C. K. Ng

    2013-04-01

    Full Text Available Successful vector-mediated plant virus transmission entails an intricate but poorly understood interplay of interactions among virus, vector, and plant. The complexity of interactions requires continually improving/evaluating tools and methods for investigating the determinants that are central to mediating virus transmission. A recent study using an organic fluorophore (Alexa Fluor-based immunofluorescent localization assay demonstrated that specific retention of Lettuce infectious yellows virus (LIYV virions in the anterior foregut or cibarium of its whitefly vector is required for virus transmission. Continuous exposure of organic fluorophore to high excitation light intensity can result in diminished or loss of signals, potentially confounding the identification of important interactions associated with virus transmission. This limitation can be circumvented by incorporation of photostable fluorescent nanocrystals, such as quantum dots (QDs, into the assay. We have developed and evaluated a QD-immunofluorescent labeling method for the in vitro and in situ localization of LIYV virions based on the recognition specificity of streptavidin-conjugated QD605 (S-QD605 for biotin-conjugated anti-LIYV IgG (B-αIgG. IgG biotinylation was verified in a blot overlay assay by probing SDS-PAGE separated B-αIgG with S-QD605. Immunoblot analyses of LIYV using B-αIgG and S-QD605 resulted in a virus detection limit comparable to that of DAS-ELISA. In membrane feeding experiments, QD signals were observed in the anterior foregut or cibarium of virion-fed whitefly vectors but absent in those of virion-fed whitefly non-vectors. Specific virion retention in whitefly vectors corresponded with successful virus transmission. A fluorescence photobleaching assay of viruliferous whiteflies fed B-αIgG and S-QD605 vs. those fed anti-LIYV IgG and Alexa Fluor 488-conjugated IgG revealed that QD signal was stable and deteriorated ∼7 to 8 fold slower than that of Alexa

  10. Kidney lesions in Rocky Mountain spotted fever: a light-, immunofluorescence-, and electron-microscopic study.

    Science.gov (United States)

    Bradford, W. D.; Croker, B. P.; Tisher, C. C.

    1979-01-01

    The essential pathologic lesion in Rocky Mountain spotted fever (RMSF) is a vasculitis that may involve the kidneys as well as the heart, brain, skin, and subcutaneous tissues. Histopathologic information concerning the response of the kidneys in RMSF is rather limited, however. In this study renal tissue from 17 children who died of RMSF was examined by light, electron, and immunofluorescence microscopy. A lymphocytic or mixed inflammation, or both, involving vessels and interstitium of the kidney was found in all patients. In addition, 10 patients had histologic evidence of acute tubular necrosis, and another 3 had glomerular lesions consisting of focal segmental tuft necrosis or increased cellularity secondary to neutophilic infiltration, or both. Immunofluorescence- and electron-microscopic studies failed to demonstrate immune-complex deposition within glomeruli, a finding that suggests that immunoglobulin and classic immune complexes were not involved in the pathogenesis of the renal lesions at the time of death. These findings suggest the possibility that the pathogenesis of the renal lesion in RMSF may be due to a direct action of the organism (Rickettsia rickettsii) on the vessel wall. Images Figure 2 Figure 1 PMID:525676

  11. Choice of Illumination System & Fluorophore for Multiplex Immunofluorescence on FFPE Tissue Sections.

    Directory of Open Access Journals (Sweden)

    Sandrine Prost

    Full Text Available The recent availability of novel dyes and alternative light sources to facilitate complex tissue immunofluorescence studies such as multiplex labelling has not been matched by reports critically evaluating the considerations and relative benefits of these new tools, particularly in combination. Product information is often limited to wavelengths used for older fluorophores (FITC, TRITC & corresponding Alexa dyes family. Consequently, novel agents such as Quantum dots are not widely appreciated or used, despite highly favourable properties including extremely bright emission, stability and potentially reduced tissue autofluorescence at the excitation wavelength. Using spectral analysis, we report here a detailed critical appraisal and comparative evaluation of different light sources and fluorophores in multiplex immunofluorescence of clinical biopsy sections. The comparison includes mercury light, metal halide and 3 different LED-based systems, using 7 Qdots (525, 565, 585, 605, 625, 705, Cy3 and Cy5. We discuss the considerations relevant to achieving the best combination of light source and fluorophore for accurate multiplex fluorescence quantitation. We highlight practical limitations and confounders to quantitation with filter-based approaches.

  12. Indirect Reciprocity; A Field Experiment.

    Science.gov (United States)

    van Apeldoorn, Jacobien; Schram, Arthur

    2016-01-01

    Indirect reciprocity involves cooperative acts towards strangers, either in response to their kindness to third parties (downstream) or after receiving kindness from others oneself (upstream). It is considered to be important for the evolution of cooperative behavior amongst humans. Though it has been widely studied theoretically, the empirical evidence of indirect reciprocity has thus far been limited and based solely on behavior in laboratory experiments. We provide evidence from an online environment where members can repeatedly ask and offer services to each other, free of charge. For the purpose of this study we created several new member profiles, which differ only in terms of their serving history. We then sent out a large number of service requests to different members from all over the world. We observe that a service request is more likely to be rewarded for those with a profile history of offering the service (to third parties) in the past. This provides clear evidence of (downstream) indirect reciprocity. We find no support for upstream indirect reciprocity (in this case, rewarding the service request after having previously received the service from third parties), however. Our evidence of downstream indirect reciprocity cannot be attributed to reputational effects concerning one's trustworthiness as a service user.

  13. Indirect Reciprocity under Incomplete Observation

    Science.gov (United States)

    Nakamura, Mitsuhiro; Masuda, Naoki

    2011-01-01

    Indirect reciprocity, in which individuals help others with a good reputation but not those with a bad reputation, is a mechanism for cooperation in social dilemma situations when individuals do not repeatedly interact with the same partners. In a relatively large society where indirect reciprocity is relevant, individuals may not know each other's reputation even indirectly. Previous studies investigated the situations where individuals playing the game have to determine the action possibly without knowing others' reputations. Nevertheless, the possibility that observers of the game, who generate the reputation of the interacting players, assign reputations without complete information about them has been neglected. Because an individual acts as an interacting player and as an observer on different occasions if indirect reciprocity is endogenously sustained in a society, the incompleteness of information may affect either role. We examine the game of indirect reciprocity when the reputations of players are not necessarily known to observers and to interacting players. We find that the trustful discriminator, which cooperates with good and unknown players and defects against bad players, realizes cooperative societies under seven social norms. Among the seven social norms, three of the four suspicious norms under which cooperation (defection) to unknown players leads to a good (bad) reputation enable cooperation down to a relatively small observation probability. In contrast, the three trustful norms under which both cooperation and defection to unknown players lead to a good reputation are relatively efficient. PMID:21829335

  14. A Novel Tool for High-Throughput Screening of Granulocyte-Specific Antibodies Using the Automated Flow Cytometric Granulocyte Immunofluorescence Test (Flow-GIFT

    Directory of Open Access Journals (Sweden)

    Xuan Duc Nguyen

    2011-01-01

    Full Text Available Transfusion-related acute lung injury (TRALI is a severe complication related with blood transfusion. TRALI has usually been associated with antibodies against leukocytes. The flow cytometric granulocyte immunofluorescence test (Flow-GIFT has been introduced for routine use when investigating patients and healthy blood donors. Here we describe a novel tool in the automation of the Flow-GIFT that enables a rapid screening of blood donations. We analyzed 440 sera from healthy female blood donors for the presence of granulocyte antibodies. As positive controls, 12 sera with known antibodies against anti-HNA-1a, -b, -2a; and -3a were additionally investigated. Whole-blood samples from HNA-typed donors were collected and the test cells isolated using cell sedimentation in a Ficoll density gradient. Subsequently, leukocytes were incubated with the respective serum and binding of antibodies was detected using FITC-conjugated antihuman antibody. 7-AAD was used to exclude dead cells. Pipetting steps were automated using the Biomek NXp Multichannel Automation Workstation. All samples were prepared in the 96-deep well plates and analyzed by flow cytometry. The standard granulocyte immunofluorescence test (GIFT and granulocyte agglutination test (GAT were also performed as reference methods. Sixteen sera were positive in the automated Flow-GIFT, while five of these sera were negative in the standard GIFT (anti—HNA 3a, n = 3; anti—HNA-1b, n = 1 and GAT (anti—HNA-2a, n = 1. The automated Flow-GIFT was able to detect all granulocyte antibodies, which could be only detected in GIFT in combination with GAT. In serial dilution tests, the automated Flow-GIFT detected the antibodies at higher dilutions than the reference methods GIFT and GAT. The Flow-GIFT proved to be feasible for automation. This novel high-throughput system allows an effective antigranulocyte antibody detection in a large donor population in order to prevent TRALI due to transfusion of

  15. A novel tool for high-throughput screening of granulocyte-specific antibodies using the automated flow cytometric granulocyte immunofluorescence test (Flow-GIFT).

    Science.gov (United States)

    Nguyen, Xuan Duc; Dengler, Thomas; Schulz-Linkholt, Monika; Klüter, Harald

    2011-02-03

    Transfusion-related acute lung injury (TRALI) is a severe complication related with blood transfusion. TRALI has usually been associated with antibodies against leukocytes. The flow cytometric granulocyte immunofluorescence test (Flow-GIFT) has been introduced for routine use when investigating patients and healthy blood donors. Here we describe a novel tool in the automation of the Flow-GIFT that enables a rapid screening of blood donations. We analyzed 440 sera from healthy female blood donors for the presence of granulocyte antibodies. As positive controls, 12 sera with known antibodies against anti-HNA-1a, -b, -2a; and -3a were additionally investigated. Whole-blood samples from HNA-typed donors were collected and the test cells isolated using cell sedimentation in a Ficoll density gradient. Subsequently, leukocytes were incubated with the respective serum and binding of antibodies was detected using FITC-conjugated antihuman antibody. 7-AAD was used to exclude dead cells. Pipetting steps were automated using the Biomek NXp Multichannel Automation Workstation. All samples were prepared in the 96-deep well plates and analyzed by flow cytometry. The standard granulocyte immunofluorescence test (GIFT) and granulocyte agglutination test (GAT) were also performed as reference methods. Sixteen sera were positive in the automated Flow-GIFT, while five of these sera were negative in the standard GIFT (anti-HNA 3a, n = 3; anti-HNA-1b, n = 1) and GAT (anti-HNA-2a, n = 1). The automated Flow-GIFT was able to detect all granulocyte antibodies, which could be only detected in GIFT in combination with GAT. In serial dilution tests, the automated Flow-GIFT detected the antibodies at higher dilutions than the reference methods GIFT and GAT. The Flow-GIFT proved to be feasible for automation. This novel high-throughput system allows an effective antigranulocyte antibody detection in a large donor population in order to prevent TRALI due to transfusion of blood products.

  16. Indirect techniques in nuclear astrophysics

    International Nuclear Information System (INIS)

    Mukhamedzhanov, A.M.; Tribble, R.E.; Blokhintsev, L.D.; Cherubini, S.; Spitaleri, C.; Kroha, V.; Nunes, F.M.

    2005-01-01

    It is very difficult or often impossible to measure in the lab conditions nuclear cross sections at astrophysically relevant energies. That is why different indirect techniques are used to extract astrophysical information. In this talk different experimental possibilities to get astrophysical information using radioactive and stable beams will be addressed. 1. The asymptotic normalization coefficient (ANC) method. 2. Radiative neutron captures are determined by the spectroscopic factors (SP). A new experimental technique to determine the neutron SPs will be addressed. 3. 'Trojan Horse' is another unique indirect method, which allows one to extract the astrophysical factors for direct and resonant nuclear reactions at astrophysically relevant energies. (author)

  17. Comparative determination of the rheumatic factor by means of agglutination, immunofluorescence and radioimmunoassay

    Energy Technology Data Exchange (ETDEWEB)

    Jaeger, L.; Storz, H.; Hein, G.; Schlenvoigt, G. (Friedrich-Schiller-Universitaet, Jena (German Democratic Republic). Bereich Medizin)

    1982-01-01

    The rheumatic factor (RF) was determined by means of agglutination, immunofluorescence (IF) test and radioimmunoassay (RIPEGA) in random groups of 56 patients with rheumatoid arthritis (RA), 13 patients with seronegative RA and 39 patients with psoriasis arthropathica. All three methods are of equal value with regard to the number of positive results. Further classification of seronegative patients, i.e. patients with a negative agglutination reaction and the clinical symptoms of RA is possible with the IF method and, above all, by means of RIPEGA. But because of the comprehensive test devices the two methods are only an alternative. Titer differences are attributed to the different indication principles and the immunological heterogeneity of RF. An improvement of the diagnosis of activity was not possible.

  18. Comparative determination of the rheumatic factor by means of agglutination, immunofluorescence and radioimmunoassay

    International Nuclear Information System (INIS)

    Jaeger, L.; Storz, H.; Hein, G.; Schlenvoigt, G.

    1982-01-01

    The rheumatic factor (RF) was determined by means of agglutination, immunofluorescence (IF) test and radioimmunoassay (RIPEGA) in random groups of 56 patients with rheumatoid arthritis (RA), 13 patients with seronegative RA and 39 patients with psoriasis arthropathica. All three methods are of equal value with regard to the number of positive results. Further classification of seronegative patients, i.e. patients with a negative agglutination reaction and the clinical symptoms of RA is possible with the IF method and, above all, by means of RIPEGA. But because of the comprehensive test devices the two methods are only an alternative. Titer differences are attributed to the different indication principles and the immunological heterogeneity of RF. An improvement of the diagnosis of activity was not possible. (author)

  19. Environmental effects of indirect subsidies

    International Nuclear Information System (INIS)

    Van Beers, C.P.; De Moor, A.P.G.; Van den Bergh, J.C.J.M.; Oosterhuis, F.H.

    2003-01-01

    The aim of the study on the title subject is to develop a transparent integrated method to determine and analyze the environmental impacts of indirect subsidies, applied in the sectors agriculture, energy, mobility, and tourism. From the results it appears that the hazardous effects of subsidies are big. Examples are milk, the regulating energy levy, and kerosene [nl

  20. Indirect searches for dark matter

    Indian Academy of Sciences (India)

    The current status of indirect searches for dark matter has been reviewed in a schematic way here. The main relevant experimental results of the recent years have been listed and the excitements and disappointments that their phenomenological interpretations in terms of almost-standard annihilating dark matter have ...

  1. Indirect methods in nuclear astrophysics

    International Nuclear Information System (INIS)

    Bertulani, C.A.; Shubhchintak; Mukhamedzhanov, A.; Kadyrov, A. S.; Kruppa, A.; Pang, D. Y.

    2016-01-01

    We discuss recent developments in indirect methods used in nuclear astrophysics to determine the capture cross sections and subsequent rates of various stellar burning processes, when it is difficult to perform the corresponding direct measurements. We discuss in brief, the basic concepts of Asymptotic Normalization Coefficients, the Trojan Horse Method, the Coulomb Dissociation Method, (d,p), and charge-exchange reactions. (paper)

  2. Indirect Reciprocity : A Field Experiment

    NARCIS (Netherlands)

    van Apeldoorn, J.; Schram, A.

    2016-01-01

    Indirect reciprocity involves cooperative acts towards strangers, either in response to their kindness to third parties (downstream) or after receiving kindness from others oneself (upstream). It is considered to be important for the evolution of cooperative behavior amongst humans. Though it has

  3. Indirect reciprocity with optional interactions.

    Science.gov (United States)

    Ghang, Whan; Nowak, Martin A

    2015-01-21

    Indirect reciprocity is a mechanism for the evolution of cooperation that is relevant for prosocial behavior among humans. Indirect reciprocity means that my behavior towards you also depends on what you have done to others. Indirect reciprocity is associated with the evolution of social intelligence and human language. Most approaches to indirect reciprocity assume obligatory interactions, but here we explore optional interactions. In any one round a game between two players is offered. A cooperator accepts a game unless the reputation of the other player indicates a defector. For a game to take place, both players must accept. In a game between a cooperator and a defector, the reputation of the defector is revealed to all players with probability Q. After a sufficiently large number of rounds the identity of all defectors is known and cooperators are no longer exploited. The crucial condition for evolution of cooperation can be written as hQB>1, where h is the average number of rounds per person and B=(b/c)-1 specifies the benefit-to-cost ratio. We analyze both stochastic and deterministic evolutionary game dynamics. We study two extensions that deal with uncertainty: hesitation and malicious gossip. Copyright © 2014 Elsevier Ltd. All rights reserved.

  4. Antismooth muscle and antiactin antibodies are indirect markers of histological and biochemical activity of autoimmune hepatitis.

    Science.gov (United States)

    Couto, Claudia A; Bittencourt, Paulo L; Porta, Gilda; Abrantes-Lemos, Clarice P; Carrilho, Flair J; Guardia, Bianca D; Cançado, Eduardo L R

    2014-02-01

    Reactivity and titers of autoantibodies vary during the course of autoimmune hepatitis (AIH), and some autoantibodies have been associated with disease activity and adverse outcomes after treatment. The aim of this study was to assess the autoantibody behavior in AIH and its significance as predictors of biochemical and histological remission. A total of 117 patients with AIH (mean age 18.6 [4-69] years) were evaluated and tested for autoantibodies at disease onset and successively (mean 3.2 [2-6] times) after a mean follow-up evaluation of 70 [20-185] months. Antismooth muscle (ASMA), antiliver kidney microsome type 1 (anti-LKM1), antiliver cytosol type 1 (anti-LC1), antimitochondrial, antinuclear (ANA), and antiactin antibodies (AAA) were determined at disease onset and 379 other times during the follow-up evaluation through indirect immunofluorescence in rodent tissues, HEp-2 cells, and human fibroblasts. Anti-SLA/LP were assessed 45 times in the follow-up evaluation of 19 patients using enzyme-linked immunosorbent assay (ELISA). Upon admission, AIH types 1 and 2 were observed in 95 and 17 patients, respectively. Five subjects had AIH with anti-SLA/LP as the sole markers. Patients initially negative for AAA did not develop these antibodies thereafter. ANA were detected de novo in six and three subjects with AIH types 1 and 2, respectively. After treatment, only ASMA (>1:80) and AAA (>1:40) were significantly associated with biochemical (76.9% and 79.8%) and histological features (100% and 100%) of disease activity (P < 0.001). With the exception of ANA, the autoantibody profile does not markedly vary in the course of AIH. The persistence of high titers of ASMA and/or AAA in patients with AIH is associated with disease activity. © 2013 by the American Association for the Study of Liver Diseases.

  5. Multiplexed Immunofluorescence Reveals Potential PD-1/PD-L1 Pathway Vulnerabilities in Craniopharyngioma.

    Science.gov (United States)

    Coy, Shannon; Rashid, Rumana; Lin, Jia-Ren; Du, Ziming; Donson, Andrew M; Hankinson, Todd C; Foreman, Nicholas K; Manley, Peter E; Kieran, Mark W; Reardon, David A; Sorger, Peter K; Santagata, Sandro

    2018-03-02

    Craniopharyngiomas are neoplasms of the sellar/parasellar region that are classified into adamantinomatous (ACP) and papillary (PCP) subtypes. Surgical resection of craniopharyngiomas is challenging, and recurrence is common, frequently leading to profound morbidity. BRAF V600E mutations render PCP susceptible to BRAF/MEK inhibitors, but effective targeted therapies are needed for ACP. We explored the feasibility of targeting the PD-1/PD-L1 immune checkpoint pathway in ACP and PCP. We mapped and quantified PD-L1 and PD-1 expression in ACP and PCP resections using immunohistochemistry, immunofluorescence, and RNA in situ hybridization. We used tissue-based cyclic immunofluorescence (t-CyCIF) to map the spatial distribution of immune cells and characterize cell cycle and signaling pathways in ACP tumor cells which intrinsically express PD-1. All ACP (15±14% of cells, n=23, average±S.D.) and PCP (35±22% of cells, n=18) resections expressed PD-L1. In ACP, PD-L1 was predominantly expressed by tumor cells comprising the cyst-lining. In PCP, PD-L1 was highly-expressed by tumor cells surrounding the stromal fibrovascular cores. ACP also exhibited tumor cell-intrinsic PD-1 expression in whorled epithelial cells with nuclear-localized beta-catenin. These cells exhibited evidence of elevated mTOR and MAPK signaling. Profiling of immune populations in ACP and PCP showed a modest density of CD8+ T-cells. ACP exhibit PD-L1 expression in the tumor cyst-lining and intrinsic PD-1 expression in cells proposed to comprise an oncogenic stem-like population. In PCP, proliferative tumor cells express PD-L1 in a continuous band at the stromal-epithelial interface. Targeting PD-L1 and/or PD-1 in both subtypes of craniopharyngioma might therefore be an effective therapeutic strategy.

  6. Manifestation of Aerosol Indirect Effects in Arctic Clouds

    Science.gov (United States)

    Lubin, D.; Vogelmann, A. M.

    2009-12-01

    The first aerosol indirect effect has traditionally been conceived as an enhancement of shortwave cloud reflectance in response to decreased effective droplet size at fixed liquid water path, as cloud nucleating aerosol becomes entrained in the cloud. The high Arctic, with its pervasive low-level stratiform cloud cover and frequent episodes of anthropogenic aerosol (Artic "haze"), has in recent years served as a natural laboratory for research on actual manifestations of aerosol indirect effects. This paper will review the surprising set of developments: (1) the detection of the indirect effect as a source of surface warming, rather than cooling, throughout early spring, (2) a transition to a cooling effect in late spring, corresponding to the beginning of the sea ice melt season, and (3) detection of an indirect effect during summer, outside of the "Arctic haze" season. This paper will also discuss measurements of spectral shortwave irradiance (350-2200 nm) made at Barrow, Alaska, during the U.S. Department of Energy's Indirect and Semi-Direct Aerosol Campaign (ISDAC), which reveal complications in our conception of the indirect effect related to the ice phase in Arctic stratiform clouds.

  7. Indirect taxation in the European Union

    OpenAIRE

    Ene, Sebastian; Micuda, Dan

    2007-01-01

    Indirect taxes are levied on the production and consumption of goods and services. They influence the retail price, and hence affect patterns of trade and consumption. Indirect taxes are ultimately paid by the final consumer. Sales and turnover taxes, excise duties and tariffs are the basic indirect taxes. In contrast with direct taxes, indirect taxes are seldom progressive. The principles for the levying of these taxes will be considered before the analysis of indirect taxes.

  8. Detection of antineutrophil cytoplasmic antibodies (ANCAs)

    DEFF Research Database (Denmark)

    Damoiseaux, Jan; Csernok, Elena; Rasmussen, Niels

    2017-01-01

    of diagnosis) from 251 patients with ANCA-associated vasculitis (AAV), including granulomatosis with polyangiitis and microscopic polyangiitis, and from 924 disease controls were tested for the presence of cytoplasmic pattern/perinuclear pattern and atypical ANCA (A-ANCA) by indirect immunofluorescence (IIF...

  9. Quantum Dots-Based Immunofluorescent Imaging of Stromal Fibroblasts Caveolin-1 and Light Chain 3B Expression and Identification of Their Clinical Significance in Human Gastric Cancer

    Directory of Open Access Journals (Sweden)

    Honglei Chen

    2012-10-01

    Full Text Available Caveolin-1 (Cav-1 expression deficiency and autophagy in tumor stromal fibroblasts (hereafter fibroblasts are involved in tumor proliferation and progression, particularly in breast and prostate cancer. The aim of this study was to detect the expression of fibroblastic Cav-1 and LC3B, markers of autophagy, in gastric cancer (GC and to analyze their clinical significances. Furthermore, because Epstein-Barr virus (EBV-associated GC (EBVaGC is a unique subtype of GC; we compared the differential expression of fibroblastic Cav-1 and LC3B in EBVaGC and non-EBVaGC. Quantum dots (QDs-based immunofluorescence histochemistry was used to examine the expression of fibroblastic Cav-1 and LC3B in 118 cases of GC with adequate stroma. QDs-based double immunofluorescence labeling was performed to detect the coexpression of Cav-1 and LC3B proteins. EBV-encoded small RNA was detected by QDs-based fluorescence in situ hybridization to identify EBVaGC. Multivariate analysis indicated that low fibroblastic Cav-1 level was an independent prognosticator (p = 0.029 that predicted poorer survival of GC patients. Positive fibroblastic LC3B was correlated with lower invasion (p = 0.032 and was positively associated with Cav-1 expression (r = 0.432, p < 0.001. EBV infection did not affect fibroblastic Cav-1 and LC3B expression. In conclusion, positive fibroblastic LC3B correlates with lower invasion, and low expression of fibroblastic Cav-1 is a novel predictor of poor GC prognosis.

  10. Electronic Commerce and Indirect Taxation

    OpenAIRE

    Watanabe, Satoshi

    2001-01-01

    This paper considers the reason why indirect tax issues arise in the context of electronic commerce and discusses relevant issues. To do so, the paper presents a simple framework to understand and examine these issues in the international context. It also identifies common and different issues of the VAT system and the RST system. It demonstrates that an emerging solution proposed in the context of the VAT system can be affected with difficult problems that were regarded to be problems of the...

  11. Indirect measurements of X-ray spectra

    International Nuclear Information System (INIS)

    Mainardi, R.T.

    2006-01-01

    To the effects of measuring the spectral distribution of the radiation emitted by the x-ray tubes and electron accelerators, numerous procedures that are grouped in two big categories exist at the present time: direct and indirect methods. The first ones use high resolution detectors that should be positioned, together with the appropriate collimator, in the direction of the x ray beam. The user should be an expert in the use and correction of the obtained data by the different effects that affect the detector operation such as efficiency and resolution in terms of the energy of the detected radiation. The indirect procedures, although its are more simple to use, its also require a considerable space along the beam to position the ionization chamber and the necessary absorbents to construct by this way the denominated attenuation curve. We will analyze the operation principle of the indirect methods and a new proposal in which such important novelties are introduced as the beam dispersion to avoid to measure along the main beam and that of determination of the attenuation curve in simultaneous form. By this way, with a single shot of the tube, the attenuation curve is measured, being necessary at most a shot of additional calibration to know the relative response of the detectors used in the experimental array. The physical processes involved in the obtaining of an attenuation curve are very well well-known and this it finishes it can be theoretically calculated if the analytic form of the spectrum is supposed well-known. Finally, we will see a spectra reconstruction example with the Kramers parametric form and comparisons with numeric simulations carried out with broadly validated programs as well as the possibility of the use of solid state dosemeters in the obtention of the attenuation curve. (Author)

  12. Segment and fit thresholding: a new method for image analysis applied to microarray and immunofluorescence data.

    Science.gov (United States)

    Ensink, Elliot; Sinha, Jessica; Sinha, Arkadeep; Tang, Huiyuan; Calderone, Heather M; Hostetter, Galen; Winter, Jordan; Cherba, David; Brand, Randall E; Allen, Peter J; Sempere, Lorenzo F; Haab, Brian B

    2015-10-06

    Experiments involving the high-throughput quantification of image data require algorithms for automation. A challenge in the development of such algorithms is to properly interpret signals over a broad range of image characteristics, without the need for manual adjustment of parameters. Here we present a new approach for locating signals in image data, called Segment and Fit Thresholding (SFT). The method assesses statistical characteristics of small segments of the image and determines the best-fit trends between the statistics. Based on the relationships, SFT identifies segments belonging to background regions; analyzes the background to determine optimal thresholds; and analyzes all segments to identify signal pixels. We optimized the initial settings for locating background and signal in antibody microarray and immunofluorescence data and found that SFT performed well over multiple, diverse image characteristics without readjustment of settings. When used for the automated analysis of multicolor, tissue-microarray images, SFT correctly found the overlap of markers with known subcellular localization, and it performed better than a fixed threshold and Otsu's method for selected images. SFT promises to advance the goal of full automation in image analysis.

  13. AMP deaminase histochemical activity and immunofluorescent isozyme localization in rat skeletal muscle

    Science.gov (United States)

    Thompson, J. L.; Sabina, R. L.; Ogasawara, N.; Riley, D. A.

    1992-01-01

    The cellular distribution of AMP deaminase (AMPda) isozymes was documented for rat soleus and plantaris muscles, utilizing immunofluorescence microscopy and immunoprecipitation methods. AMPda is a ubiquitous enzyme existing as three distinct isozymes, A, B and C, which were initially purified from skeletal muscle, liver (and kidney), and heart, respectively. AMPda-A is primarily concentrated subsarcolemmally and intermyofibrillarly within muscle cells, while isozymes B and C are concentrated within non-myofiber elements of muscle tissue. AMPda-B is principally associated with connective tissues surrounding neural elements and the muscle spindle capsule, and AMPda-C is predominantly associated with circulatory elements, such as arterial and venous walls, capillary endothelium, and red blood cells. These specific localizations, combined with documented differences in kinetic properties, suggest multiple functional roles for the AMPda isozymes or temporal segregation of similar AMPda functions. Linkage of the AMPda substrate with adenosine production pathways at the AMP level and the localization of isozyme-C in vascular tissue suggest a regulatory role in the microcirculation.

  14. Pattern of glomerular diseases in oman: A study based on light microscopy and immunofluorescence

    Directory of Open Access Journals (Sweden)

    Nasar Yousuf Alwahaibi

    2013-01-01

    Full Text Available Light microscopy and immunofluorescence play an important part in the final diagnosis of renal biopsy. The aim of this study was to analyze the pattern of various glomerular diseases in Oman. A total of 424 renal biopsies were retrospectively analyzed at the Sultan Qaboos University Hospital between 1999 and 2010. Focal and segmental glomerulosclerosis (FSGS, minimal change disease (MCD, membranous glomerulopathy (MGN and IgA nephropathy were the most common primary glomerular diseases encountered, accounting for 21.2%, 17%, 12.3% and 8.3%, respectively, of all cases. Lupus nephritis was the most common secondary glomerular disease and was the most prevalent among all biopsies, accounting for 30.4% of all biopsies. Amyloidosis was seen in only two cases. The presence of fluorescein isothiocyanatefibrin in all renal cases was low when compared with IgG, IgA, IgM, C3 and C1q markers. In conclusion, based on the findings of this study, lupus nephritis was the most common of all glomerular diseases and FSGS was the most common primary glomerular disease. The importance of fluorescein isothiocyanate-fibrin in the diagnosis of renal biopsy needs to be further investigated.

  15. Detection of anti-Giardia lamblia serum antibody among children of day care centers

    Directory of Open Access Journals (Sweden)

    Guimarães Semíramis

    2002-01-01

    Full Text Available OBJETIVES: To detect anti-Giardia lamblia serum antibodies in healthy children attending public day care centers and to assess serological tests as tools for estimating the prevalence of G. lamblia in endemic areas. METHODS: Three separate stool specimens and filter paper blood samples were collected from 147 children ranging from 0 to 6 years old. Each stool sample was processed using spontaneous sedimentation and zinc sulfate flotation methods. Blood samples were tested by indirect immunofluorescence (IIF and enzyme-linked immunosorbent assay (ELISA for Giardia IgG. RESULTS AND CONCLUSIONS: Of 147 individuals tested, 93 (63.3% showed Giardia cysts in their feces. Using IIF and ELISA, serum antibodies were detected in 93 (63.3% and 100 (68% samples , respectively. Sensitivity of IIF and ELISA was 82% and 72%, respectively. However, ELISA revealed to be less specific (39% than IIF (70%. IIF also showed a higher concordance with microscopic examination than ELISA.

  16. Characterization of pars intermedia connections in amphibians by biocytin tract tracing and immunofluorescence aided by confocal microscopy

    NARCIS (Netherlands)

    Jansen, K; Fabro, C; Artero, C; Feuilloley, M; Vaudry, H; Fasolo, A; Franzoni, MF

    Biocytin, recently introduced in neuroanatomical studies, was used as a retrograde tract tracer in combination with immunofluorescence in order to analyse the neurochemical characters of some central neuronal projections to the pars intermedia in two amphibian species, the anuran Rana esculenta and

  17. Immunofluorescence-based screening identifies germ cell associated microRNA 302 as an antagonist to p63 expression

    DEFF Research Database (Denmark)

    Scheel, Andreas Hans Joachim; Beyer, Ulrike; Agami, Reuven

    2009-01-01

    The tumor suppressor homologue p63 is required for proper skin and limb development, but specific isoforms of it also act as a "guardian of the germline." To gain insight into the regulation of p63 expression, we performed immunofluorescence-based screening assays. Using a large collection of micro...

  18. Indirect Medical Education and Disproportionate Share Adj...

    Data.gov (United States)

    U.S. Department of Health & Human Services — Indirect Medical Education and Disproportionate Share Adjustments to Medicare Inpatient Payment Rates The indirect medical education (IME) and disproportionate share...

  19. Indirect costs of rheumatoid arthritis

    Directory of Open Access Journals (Sweden)

    Filip Raciborski

    2015-12-01

    Full Text Available It is estimated that in Poland about 400,000 persons in general suffer from inflammatory joint diseases, including rheumatoid arthritis (RA. Epidemiological surveys documenting the frequency and disturbance of musculoskeletal disorders in the Polish population are few in number. Most of the estimations are based on epidemiological data from other countries (prevalence of 0.5–1%. According to the data of the National Health Fund in Poland 135,000–157,000 persons in total are treated because of rheumatoid arthritis per year [ICD10 (International Statistical Classification of Diseases and Related Health Problems: M05, M06]. In the case of this group of diseases indirect costs significantly outweigh the direct costs. Indirect costs increase together with activity level of the disease. The cost analysis of productivity loss of RA patients indicates that sickness absenteeism and informal care are the most burdensome. At the national level it amounts in total from 1.2 billion to 2.8 billion PLN per year, depending on the method of analysis. These costs could be significantly reduced through early diagnosis and introduction of effective treatment.

  20. Indirect reciprocity with trinary reputations.

    Science.gov (United States)

    Tanabe, Shoma; Suzuki, Hideyuki; Masuda, Naoki

    2013-01-21

    Indirect reciprocity is a reputation-based mechanism for cooperation in social dilemma situations when individuals do not repeatedly meet. The conditions under which cooperation based on indirect reciprocity occurs have been examined in great details. Most previous theoretical analysis assumed for mathematical tractability that an individual possesses a binary reputation value, i.e., good or bad, which depends on their past actions and other factors. However, in real situations, reputations of individuals may be multiple valued. Another puzzling discrepancy between the theory and experiments is the status of the so-called image scoring, in which cooperation and defection are judged to be good and bad, respectively, independent of other factors. Such an assessment rule is found in behavioral experiments, whereas it is known to be unstable in theory. In the present study, we fill both gaps by analyzing a trinary reputation model. By an exhaustive search, we identify all the cooperative and stable equilibria composed of a homogeneous population or a heterogeneous population containing two types of players. Some results derived for the trinary reputation model are direct extensions of those for the binary model. However, we find that the trinary model allows cooperation under image scoring under some mild conditions. Copyright © 2012 Elsevier Ltd. All rights reserved.

  1. Indirect Cost Reimbursement: An Industrial View.

    Science.gov (United States)

    Bolton, Robert

    1987-01-01

    The meaning of indirect costs in an industrial environment is discussed. Other factors considered are corporate policies; nature of work being supported; the uniqueness of the work; who is doing the negotiating for industry; and indirect rates. Suggestions are offered for approaches to indirect cost reimbursement. (Author/MLW)

  2. Language shifts in free indirect discourse

    NARCIS (Netherlands)

    Maier, Emar

    Free indirect discourse is a way of reporting what a protagonist thinks or says that is distinct from both direct and indirect discourse. In particular, while pronouns and tenses are presented from the narrator's perspective, as in indirect discourse, other indexical and expressive elements reflect

  3. Directe en indirecte werknemersparticipatie in Europa

    NARCIS (Netherlands)

    van Houten, Gijs; Akkerman, Agnes; Sluiter, Roderick; Jansen, Giedo; Vermeylen, Greet

    2016-01-01

    This study looks at different forms of direct and indirect employee participation in the EU. The research questions are: (1) which forms of direct and indirect employee participation can we distinguish?; (2) to what extent do forms of direct and indirect employee participation coincide within

  4. Determination of cutoff of ELISA and immunofluorescence assay for scrub typhus

    Directory of Open Access Journals (Sweden)

    Nitin Gupta

    2016-01-01

    Full Text Available Background: The most common method employed for diagnosis of scrub typhus is serology. It is widely known that demonstration of ≥4-fold rise in titers of antibody in paired sera is required for diagnosis. However, for guidance of initial treatment, there is a need for rapid diagnosis at the time of admission. Therefore, there is a need for standardized region specific cutoff titers at the time of admission. Materials and Methods: A total of 258 patients of all age groups with clinically suspected scrub typhus over a period of 24 months (October 2013-October 2015 were enrolled. Serum samples of these patients were subjected to immunofluorescent antibody (IFA for immunoglobulin M (IgM (Fuller Labs, USA with dilutions of 1:64, 1:128, 1:256, and 1:512. Serum samples of all 258 patients were subjected to IgM ELISA (Inbios Inc., USA. Any patient with response to antibiotics within 48 h accompanied by either presence of an eschar or positivity by polymerase chain reaction was taken as positive. Receiver operating characteristic (ROC curve was drawn to generate cutoff for these tests. Results: A total of 20 patients were diagnosed as cases of scrub typhus. The ROC curve analysis revealed a cutoff optical density value of 0.87 with sensitivity and specificity of 100% and 94.12%, respectively. ROC curve analysis of IFA revealed sensitivity and specificity of 100% and 93.5%, respectively at 1:64 dilution. Conclusion: Considering cost constraints, centers in and around New Delhi region can use the cutoffs we determined for the diagnosis of scrub typhus.

  5. Indirect estimation of radioactivity in containerized cargo

    International Nuclear Information System (INIS)

    Jarman, K.D.; Scherrer, C.; Smith, L.E.; Chilton, L.K.; Anderson, K.K.; Ressler, J.J.; Trease, L.L.

    2011-01-01

    Naturally occurring radioactive material in containerized cargo challenges the state of the art in national and international efforts to detect illicit nuclear and radiological material in transported containers. Current systems are being evaluated and new systems envisioned to provide the high probability of detection necessary to thwart potential threats, combined with extremely low nuisance and false alarm rates necessary to maintain the flow of commerce impacted by the enormous volume of commodities imported in shipping containers. Maintaining flow of commerce also means that inspection must be rapid, requiring relatively non-intrusive, indirect measurements of cargo from outside containers to the extent possible. With increasing information content in such measurements, it is natural to ask how the information might be combined to improve detection. Toward this end, we present an approach to estimating isotopic activity of naturally occurring radioactive material in cargo grouped by commodity type, combining container manifest data with radiography and gamma-ray spectroscopy aligned to location along the container. The heart of this approach is our statistical model of gamma-ray counts within peak regions of interest, which captures the effects of background suppression, counting noise, convolution of neighboring cargo contributions, and down-scattered photons to provide estimates of counts due to decay of specific radioisotopes in cargo alone. Coupled to that model, we use a mechanistic model of self-attenuated radiation flux to estimate the isotopic activity within cargo, segmented by location within each container, that produces those counts. We test our approach by applying it to a set of measurements taken at the Port of Seattle in 2006. This approach to synthesizing disparate available data streams and extraction of cargo characteristics, while relying on several simplifying assumptions and approximations, holds the potential to support improvement of

  6. Detection of intracellular canine distemper virus antigen in mink inoculated with an attenuated or a virulent strain of canine distemper virus.

    Science.gov (United States)

    Blixenkrone-Møller, M

    1989-09-01

    Using an indirect immunofluorescence technique, the distribution of viral antigen in various tissues and blood mononuclear leukocytes was studied in wild mink, either vaccinated with an attenuated vaccine strain of canine distemper virus (CDV) or experimentally inoculated with the virulent Snyder-Hill strain of CDV. Viral antigen was detected in cells of the lymphoid system 6 to 12 days after vaccination. From 2 to 3 days after inoculation with the virulent strain, CDV antigen was demonstrated in cells of the lymphoid system and, during the incubation period, the antigen had spread to the epithelia and brain at days 6 and 12, respectively. In clinical cases of acute fatal canine distemper, the viral antigen was detected in a wide variety of tissues, including the cells of the lymphoid system, epithelial cells of skin, mucous membranes, lung, kidney, and cells of the CNS. The diagnostic importance of CDV antigen detection is discussed on the basis of these findings.

  7. Using immunoglobulin Y as an alternative antibody for the detection of hepatitis A virus in frozen liver sections

    Directory of Open Access Journals (Sweden)

    Gentil Arthur Bentes

    2015-06-01

    Full Text Available An increasing amount of research has been conducted on immunoglobulin Y (IgY because the use of IgY offers several advantages with respect to diagnostic testing, including its easy accessibility, low cost and translatability to large-scale production, in addition to the fact that it can be ethically produced. In a previous work, immunoglobulin was produced and purified from egg yolks (IgY reactive to hepatitis A virus (HAV antigens. In the present work, this anti-HAV-specific IgY was used in an indirect immunofluorescence assay to detect viral antigens in liver biopsies that were obtained from experimentally infected cynomolgus monkeys. Fields that were positive for HAV antigen were detected in liver sections using confocal microscopy. In conclusion, egg yolks from immunised hens may be a reliable source for antibody production, which can be employed for immunological studies.

  8. Indirect Voltammetric Sensing Platforms For Fluoride Detection on Boron-Doped Diamond Electrode Mediated via [FeF6]3− and [CeF6]2− Complexes Formation

    International Nuclear Information System (INIS)

    Culková, Eva; Tomčík, Peter; Švorc, Ľubomír; Cinková, Kristína; Chomisteková, Zuzana; Durdiak, Jaroslav; Rievaj, Miroslav; Bustin, Dušan

    2014-01-01

    Very simple and sensitive electroanalytical technique based on synergistic combination of reaction electrochemistry (specificity) and bare boron-doped diamond electrode (sensitivity) for the detection of fluorides in drinking water was developed as variant based on dynamic electrochemistry to ISE analysis. It is based on the formation of electroinactive fluoride complexes with Fe(III) and Ce(IV) ions decreasing such diffusion current of free metal on boron-doped diamond electrode. Due to low background signal of boron-doped diamond electrode reasonably low detection limits of the order of 10 −6 mol L −1 for linear sweep voltammetric method using formation of [FeF 6 ] 3− and 10 −7 mol L −1 in a square-wave variant of this technique have been achieved. This is approximately 1–2 orders lower than in the case of platinum comb-shaped interdigitated microelectrode array. Linear sweep voltammetric method based on [CeF 6 ] 2− complex formation has lower sensitivity and may be suitable for samples with higher content of fluoride and not to analysis of drinking water

  9. Anticuerpos antinucleares, imágenes y características obtenidas por inmunofluorescencia: Importancia de los isotipos IgA, IgM e IgG Antinuclear antibodies, patterns and characteristics obtained by immunofluorescence: The importance of the IgA, IgM and IgG isotypes

    Directory of Open Access Journals (Sweden)

    Miriam Arcavi

    2009-10-01

    Full Text Available La técnica de elección para el screening de anticuerpos antinucleares (ANA es la inmunofluorescencia indirecta que utiliza como sustrato una línea de células epiteliales de carcinoma de laringe humano (IFI-HEp2, y como antisuero, anti-IgG o anti-Ig totales. Los ANA-IgG son los más importantes para el diagnóstico y monitoreo de las enfermedades del tejido conectivo (ETC, mientras los ANA-IgM son de menor relevancia clínica en estos pacientes. Sin embargo, poco se sabe de los ANA-IgA ya que estos Ac han sido menos investigados. El objetivo de este trabajo fue estudiar la prevalencia de los diferentes isotipos de inmunoglobulinas de anticuerpos antinucleares en los pacientes con ETC y evaluar la conveniencia de utilizar conjugados monovalentes o polivalentes. Se procesaron 100 sueros de pacientes con diversas ETC empleando IFI-HEp2, en los cuales se detectó 38% de ANA-IgA (títulos ≥ 1:80 y 12% de ANA-IgM (títulos ≤ 1:160. En 29 casos se detectó IgA en ausencia de IgM, en 3 casos IgM en ausencia de IgA. En todos los casos los ANA-IgG estuvieron presentes. En 6 sueros se observó un cambio de imagen con conjugado anti-IgA y en 3 con conjugado anti-IgM. Debido a la alta prevalencia de ANA-IgA detectada por IFI-HEp2, se destaca la conveniencia de utilizar conjugado anti-Ig totales en lugar de anti-IgG, mientras se desconozca la relevancia de los ANA-IgA en el diagnóstico, pronóstico y seguimiento de las enfermedades reumáticas sistémicas.The indirect immunofluorescence with epitelial cell line from human laryngeal carcinoma as substrate (IIF-HEp2 and anti-IgG or anti-total Ig as antisera, is the technique currently used for the detection of antinuclear antibodies. The most important antibodies for the diagnosis and follow-up of connective tissue diseases (CTD are the IgG-ANA, while the IgM-ANA have no clinical relevance. However the IgA-ANA have not been thoroughly investigated so far. The aim of this work was to study the prevalence

  10. Indirect Self-Destructiveness and Emotional Intelligence.

    Science.gov (United States)

    Tsirigotis, Konstantinos

    2016-06-01

    While emotional intelligence may have a favourable influence on the life and psychological and social functioning of the individual, indirect self-destructiveness exerts a rather negative influence. The aim of this study has been to explore possible relations between indirect self-destructiveness and emotional intelligence. A population of 260 individuals (130 females and 130 males) aged 20-30 (mean age of 24.5) was studied by using the Polish version of the chronic self-destructiveness scale and INTE, i.e., the Polish version of the assessing emotions scale. Indirect self-destructiveness has significant correlations with all variables of INTE (overall score, factor I, factor II), and these correlations are negative. The intensity of indirect self-destructiveness differentiates significantly the height of the emotional intelligence and vice versa: the height of the emotional intelligence differentiates significantly the intensity of indirect self-destructiveness. Indirect self-destructiveness has negative correlations with emotional intelligence as well as its components: the ability to recognize emotions and the ability to utilize emotions. The height of emotional intelligence differentiates the intensity of indirect self-destructiveness, and vice versa: the intensity of indirect self-destructiveness differentiates the height of emotional intelligence. It seems advisable to use emotional intelligence in the prophylactic and therapeutic work with persons with various types of disorders, especially with the syndrome of indirect self-destructiveness.

  11. Serological and molecular detection of Theileria equi in sport horses of northeastern Brazil.

    Science.gov (United States)

    Ferreira, Edlainne P; Vidotto, Odilon; Almeida, Jonatas C; Ribeiro, Luana P S; Borges, Marcos V; Pequeno, Walter H C; Stipp, Danilo T; de Oliveira, Celso J B; Biondo, Alexander W; Vieira, Thállitha S W J; Vieira, Rafael F C

    2016-08-01

    Theileriosis is a worldwide protozoal tick-borne disease caused by Theileria equi, which may produce a variety of clinical signs and turn infected horses into lifetime carriers. This study has aimed to perform a serological and molecular detection of T. equi and associated factors in sports horses from six areas of northeastern Brazil. In overall, 59.6% horses were positive by indirect immunofluorescence assay and 50.4% by polymerase chain reaction. No significant association was found when presence of ticks, age, gender, anemia or total plasma proteins was analyzed with seropositivity and molecular techniques. Although a significant association of infection was found in two cities. Thus, local risk factors other than presence of ticks, horse age, gender, anemia and total plasmatic proteins may dictate prevalence of T. equi infection in sports horses, even in highly endemic areas with no control of infection prior to horse competitions. Copyright © 2016 Elsevier Ltd. All rights reserved.

  12. Detection of virulence factors and molecular typing of pathogenic Leptospira from capybara (Hydrochaeris hydrochaeris).

    Science.gov (United States)

    Jorge, Sérgio; Monte, Leonardo G; Coimbra, Marco Antonio; Albano, Ana Paula; Hartwig, Daiane D; Lucas, Caroline; Seixas, Fabiana K; Dellagostin, Odir A; Hartleben, Cláudia P

    2012-10-01

    Leptospirosis is a globally prevalent zoonosis caused by pathogenic Leptospira spp.; several serologic variants have reservoirs in synanthropic rodents. The capybara is the largest living rodent in the world, and it has a wide geographical distribution in Central and South America. This rodent is a significant source of Leptospira since the agent is shed via urine into the environment and is a potential public health threat. In this study, we isolated and identified by molecular techniques a pathogenic Leptospira from capybara in southern Brazil. The isolated strain was characterized by partial rpoB gene sequencing and variable-number tandem-repeats analysis as L. interrogans, serogroup Icterohaemorrhagiae. In addition, to confirm the expression of virulence factors, the bacterial immunoglobulin-like proteins A and B expression was detected by indirect immunofluorescence using leptospiral specific monoclonal antibodies. This report identifies capybaras as an important source of infection and provides insight into the epidemiology of leptospirosis.

  13. Elucidating the Burden of HIV in Tissues Using Multiplexed Immunofluorescence and In Situ Hybridization: Methods for the Single-Cell Phenotypic Characterization of Cells Harboring HIV In Situ.

    Science.gov (United States)

    Vasquez, Joshua J; Hussien, Rajaa; Aguilar-Rodriguez, Brandon; Junger, Henrik; Dobi, Dejan; Henrich, Timothy J; Thanh, Cassandra; Gibson, Erica; Hogan, Louise E; McCune, Joseph; Hunt, Peter W; Stoddart, Cheryl A; Laszik, Zoltan G

    2018-02-01

    Persistent tissue reservoirs of HIV present a major barrier to cure. Defining subsets of infected cells in tissues is a major focus of HIV cure research. Herein, we describe a novel multiplexed in situ hybridization (ISH) (RNAscope) protocol to detect HIV-DNA (vDNA) and HIV-RNA (vRNA) in formalin-fixed paraffin-embedded (FFPE) human tissues in combination with immunofluorescence (IF) phenotyping of the infected cells. We show that multiplexed IF and ISH (mIFISH) is suitable for quantitative assessment of HIV vRNA and vDNA and that multiparameter IF phenotyping allows precise identification of the cellular source of the ISH signal. We also provide semi-quantitative data on the impact of various tissue fixatives on the detectability of vDNA and vRNA with RNAscope technology. Finally, we describe methods to quantitate the ISH signal on whole-slide digital images and validation of the quantitative ISH data with quantitative real-time PCR for vRNA. It is our hope that this approach will provide insight into the biology of HIV tissue reservoirs and to inform strategies aimed at curing HIV.

  14. Synchronization of indirectly coupled Lorenz oscillators

    Indian Academy of Sciences (India)

    Synchronization of indirectly coupled Lorenz oscillators: An experimental study. Amit Sharma Manish Dev Shrimali. Synchronization, Coupled Systems and Networks Volume 77 Issue 5 November 2011 pp 881-889 ... The in-phase and anti-phase synchronization of indirectly coupled chaotic oscillators reported in Phys. Rev ...

  15. Indirect search for dark matter with AMS

    International Nuclear Information System (INIS)

    Goy, Corinne

    2006-01-01

    This document summarises the potential of AMS in the indirect search for Dark Matter. Observations and cosmology indicate that the Universe may include a large amount of Dark Matter of unknown nature. A good candidate is the Ligthest Supersymmetric Particle in R-Parity conserving models. AMS offers a unique opportunity to study Dark Matter indirect signature in three spectra: gamma, antiprotons and positrons

  16. Indirectness in Discourse: Ethnicity as Conversational Style.

    Science.gov (United States)

    Tannen, Deborah

    1981-01-01

    Examines social differences in expectations of indirectness in conversations between married couples, both Greek and Greek-American. Concludes that Greeks are more likely to expect indirectness in the context presented and that Greek-Americans have retained the Greek communicative patterns. (FL)

  17. JUSTIFICATION FOR INDIRECT DISCRIMINATION IN EU

    Directory of Open Access Journals (Sweden)

    Cătălina-Adriana Ivănuş

    2014-11-01

    Full Text Available The right to non-discrimination is very important for a civilized society. EU legislation establishes direct and indirect discrimination, harassment, sexual harassment, instruction to discriminate and any less favourable treatment of a woman related to pregnancy or maternity leave as forms of discrimination. The law and the Court of Justice permit the justification of indirect discrimination.

  18. 77 FR 41899 - Indirect Food Additives: Polymers

    Science.gov (United States)

    2012-07-17

    ... DEPARTMENT OF HEALTH AND HUMAN SERVICES Food and Drug Administration 21 CFR Part 177 [Docket No. FDA-2012-F-0031] Indirect Food Additives: Polymers AGENCY: Food and Drug Administration, HHS. ACTION... CFR part 177 is amended as follows: PART 177--INDIRECT FOOD ADDITIVES: POLYMERS 0 1. The authority...

  19. JUSTIFICATION FOR INDIRECT DISCRIMINATION IN EU

    OpenAIRE

    Cătălina-Adriana Ivănuş

    2014-01-01

    The right to non-discrimination is very important for a civilized society. EU legislation establishes direct and indirect discrimination, harassment, sexual harassment, instruction to discriminate and any less favourable treatment of a woman related to pregnancy or maternity leave as forms of discrimination. The law and the Court of Justice permit the justification of indirect discrimination.

  20. Indirect techniques in nuclear astrophysics: a review.

    Science.gov (United States)

    Tribble, R E; Bertulani, C A; Cognata, M La; Mukhamedzhanov, A M; Spitaleri, C

    2014-10-01

    In this review, we discuss the present status of three indirect techniques that are used to determine reaction rates for stellar burning processes, asymptotic normalization coefficients, the Trojan Horse method and Coulomb dissociation. A comprehensive review of the theory behind each of these techniques is presented. This is followed by an overview of the experiments that have been carried out using these indirect approaches.

  1. Indirect Calorimetry in Mechanically Ventilated Patients

    DEFF Research Database (Denmark)

    Allingstrup, Matilde Jo; Kondrup, Jens; Perner, Anders

    2017-01-01

    Background and Aims: The 2 currently available indirect calorimeters, CCM Express Indirect Calorimeter (MedGraphics, St Paul, MN) and Quark RMR ICU Indirect Calorimeter (COSMED, Rome, Italy), have not been validated against a gold standard in mechanically ventilated patients. Our aim was to do so...... using a gold-standard, modified Tissot bell-spirometer method in mechanically ventilated patients who were hemodynamically, respiratory, and metabolically stable. Methods: We studied 30 patients undergoing general anesthesia and major gynecological surgery. We measured oxygen consumption ((Formula...... of 77 (167) with limits of agreement −249 to 404 kcal/d. Conclusions: The QUARK RMR ICU Indirect Calorimeter compared better with the gold standard for values of (Formula presented.) O2 and REE than did the CCM Express Indirect Calorimeter in mechanically ventilated patients who were circulatory...

  2. Development of polyclonal antibodies for the detection of ...

    African Journals Online (AJOL)

    2013-09-11

    month-old male New Zealand rabbits were immunized using .... Immunofluorescence analysis of anti-rHuEPO pAb in CHO cells transfected with pTARGET/EPO. ... erythropoietin gene doping: detection strategies in the genomic era.

  3. Comparison of indirect ELISA based on recombinant protein NcSRS2 and IFAT for detection of Neospora caninum antibodies in sheep Comparação entre ELISA baseado no antígeno recombinante NcSRS2 e RIFI para detecção de anticorpos de Neospora caninum em ovinos

    Directory of Open Access Journals (Sweden)

    Renato Andreotti

    2009-06-01

    Full Text Available Neospora caninum, an Apicomplexan parasite that can causes abortion, is responsible for considerable economic and reproductive losses in livestock. The purpose of the present study was to determine whether recombinant NcSRS2 is a suitable indirect ELISA antigen for determining specific immune response to N. caninum in sheep. A total of 441 serum samples were subjected to IFAT and rNcSRS2 based-ELISA, with both tests performing similarly. The sensitivity and specificity of indirect ELISA were 98.6 and 98.3%, respectively. The kappa index shows 0.98 concordance between the two tests, which is considered excellent. Seroprevalences of 30.8 and 32.0% were detected by IFAT and indirect ELISA, respectively, showing these tests did not differ significantly on this measure (p > 0.05. Serological analysis showed that HisG tag was detected by Western Blotting recognizing rNcSRS2 protein. The potential value of rNcSRS2-based ELISA as a highly specific and sensitive tool for serological diagnosis is also supported by the strong agreement found between IFAT and ELISA. The results support the potential use of recombinant protein NcSRS2 as an antigen in indirect ELISA in sheep.Neospora caninum é um parasito Apicomplexa que pode causar abortos e é reconhecido como agente importante responsável por perdas econômicas e reprodutivas. Este estudo avaliou a proteína recombinante NcSRS2 como antígeno para ELISA indireto na determinação de resposta imune para N. caninum em ovinos. 441 amostras de soro foram analisadas por IFAT e ELISA indireto com rNcSRS2 e ambos os testes revelaram comportamento similar. A sensibilidade e especificidade de ELISA indireto foram 98,6 e 98,3%, respectivamente. O índice kappa mostrou uma concordância entre os dois testes com valor de 0,98, que é considerado excelente. Prevalências de 30,8 e 32,0% detectadas por IFAT e ELISA indireto, respectivamente, mostraram que os testes não diferiram significativamente nesse aspecto (P

  4. Indirect Tire Monitoring System - Machine Learning Approach

    Science.gov (United States)

    Svensson, O.; Thelin, S.; Byttner, S.; Fan, Y.

    2017-10-01

    The heavy vehicle industry has today no requirement to provide a tire pressure monitoring system by law. This has created issues surrounding unknown tire pressure and thread depth during active service. There is also no standardization for these kind of systems which means that different manufacturers and third party solutions work after their own principles and it can be hard to know what works for a given vehicle type. The objective is to create an indirect tire monitoring system that can generalize a method that detect both incorrect tire pressure and thread depth for different type of vehicles within a fleet without the need for additional physical sensors or vehicle specific parameters. The existing sensors that are connected communicate through CAN and are interpreted by the Drivec Bridge hardware that exist in the fleet. By using supervised machine learning a classifier was created for each axle where the main focus was the front axle which had the most issues. The classifier will classify the vehicles tires condition and will be implemented in Drivecs cloud service where it will receive its data. The resulting classifier is a random forest implemented in Python. The result from the front axle with a data set consisting of 9767 samples of buses with correct tire condition and 1909 samples of buses with incorrect tire condition it has an accuracy of 90.54% (0.96%). The data sets are created from 34 unique measurements from buses between January and May 2017. This classifier has been exported and is used inside a Node.js module created for Drivecs cloud service which is the result of the whole implementation. The developed solution is called Indirect Tire Monitoring System (ITMS) and is seen as a process. This process will predict bad classes in the cloud which will lead to warnings. The warnings are defined as incidents. They contain only the information needed and the bandwidth of the incidents are also controlled so incidents are created within an

  5. ICARE improves antinuclear antibody detection by overcoming the barriers preventing accreditation.

    Science.gov (United States)

    Bertin, Daniel; Mouhajir, Yassin; Bongrand, Pierre; Bardin, Nathalie

    2016-02-15

    Antinuclear antibodies (ANA) are useful biomarkers for the diagnosis and the monitoring of rheumatic diseases. The American College of Rheumatology has stated that indirect immunofluorescence (IIF) analysis remains the gold standard for ANA screening. However, IIF is time consuming, subjective, not fully standardized and presents several issues for accreditation which is the process leading to ISO 15189 certification for medical laboratories. We propose an innovative tool for accreditation by using the quantitative evaluation of the automated image capture and analysis "ICARE" (Immunofluorescence for Computed Antinuclear antibody Rational Evaluation). We established the optimal screening dilution (1:160) and a fluorescence index (FI) cutoff for ICARE on a cohort of 91 healthy blood donors. Then, we evaluated performance of ICARE on a routine cohort of 236 patients. Precision parameters of ANA detection by IIF were evaluated according to ISO 15189. ICARE showed an excellent concordance with visual evaluation (88%, Kappa=0.76) and significantly discriminated between weak to moderate (1:160-1:320 titers) and high (>1:320 titers) ANA levels. A significant correlation was found between FI and ANA titers (Spearman's ρ=0.67; Pprocess of continuous improvement of the quality of clinical laboratories. Copyright © 2015 Elsevier B.V. All rights reserved.

  6. Quantum-dot-based immunofluorescent imaging of HER2 and ER provides new insights into breast cancer heterogeneity

    International Nuclear Information System (INIS)

    Chen Chuang; Li Yan; Peng Jun; Xu Hao; Tang Hongwu; Zhang Zhiling; Pang Daiwen; Xia Heshun; Wu Qiongshui; Zeng Libo; Zhu Xiaobo

    2010-01-01

    Breast cancer (BC) is a heterogeneous tumor, and better understanding of its heterogeneity is essential to improving treatment effect. Quantum dot (QD)-based immunofluorescent nanotechnology (QD-IHC) for molecular pathology has potential advantages in delineating tumor heterogeneity. This potential is explored in this paper by QD-IHC imaging of HER2 and ER. BC heterogeneity can be displayed more clearly and sensitively by QD-IHC than conventional IHC in BC tissue microarrays. Furthermore, the simultaneous imaging of ER and HER2 might help understand their interactions during the process of evolution of heterogeneous BC.

  7. Exploitation of immunofluorescence for the quantification and characterization of small numbers of Pasteuria endospores.

    Science.gov (United States)

    Costa, Sofia R; Kerry, Brian R; Bardgett, Richard D; Davies, Keith G

    2006-12-01

    The Pasteuria group of endospore-forming bacteria has been studied as a biocontrol agent of plant-parasitic nematodes. Techniques have been developed for its detection and quantification in soil samples, and these mainly focus on observations of endospore attachment to nematodes. Characterization of Pasteuria populations has recently been performed with DNA-based techniques, which usually require the extraction of large numbers of spores. We describe a simple immunological method for the quantification and characterization of Pasteuria populations. Bayesian statistics were used to determine an extraction efficiency of 43% and a threshold of detection of 210 endospores g(-1) sand. This provided a robust means of estimating numbers of endospores in small-volume samples from a natural system. Based on visual assessment of endospore fluorescence, a quantitative method was developed to characterize endospore populations, which were shown to vary according to their host.

  8. Expression of future prospective in indirect speech

    Directory of Open Access Journals (Sweden)

    Bodnaruk Elena Vladimirovna

    2015-03-01

    Full Text Available The article analyzes the characteristics and use of grammatical semantics and lexical and grammatical means used to create future prospects in double indirect discourse. The material for the study were epic works by contemporary German writers. In the analysis of the empirical material it has been pointed out that indirect discourse has preterial basis and is the kind of most frequent inner speech of characters. The most widely used form with future semantics in preterial indirect speech is conditional I, formally having a conjunctive basis, but is mostly used with the indicative semantics. Competitive to conditional I in indirect speech is preterial indicative. A characteristic feature of the indirect speech is the use of modal verbs, which, thanks to its semantics is usually referred as an action at a later term, creating the prospect of future statements. The most frequent were modal verbs wollen and sollen in the form of the preterite, more rare verbs were m ssen and k nnen. German indirect speech distinguishes the ability to use forms on the basis of conjunctive: preterite and plusquamperfect of conjunctive. Both forms express values similar to those of the indicative. However, conjunctive forms the basis of the data shown in a slightly more pronounced seme of uncertainty that accompanies future uses of these forms in indirect speech. In addition, plusquamperfect conjunctive differs from others by the presence of the seme of completeness.

  9. IgA anti-Actin antibodies in children with celiac disease: comparison of immunofluorescence with Elisa assay in predicting severe intestinal damage

    Directory of Open Access Journals (Sweden)

    Mora Stefano

    2010-03-01

    Full Text Available Abstract Background Previous studies have demonstrated that the presence of serum IgA antibodies against actin filaments (AAA in patients with celiac disease (CD is strongly associated with mucosal damage and severe degrees of villous atrophy. The aims of the present study were (1 to verify the effectiveness of IgA-AAA in newly diagnosed CD patients in a clinical setting (2 to compare the immunofluorescence assay with ELISA assay; (3 to compare the correlation of our IgA anti-tissue transglutaminase antibodies (tTG-Ab class with mucosal intestinal lesions. Methods 90 patients underwent endoscopy and multiple biopsies for suspected CD on the basis of symptoms, in presence of positive tTG-Ab tests. Twenty biopsied and 25 not-biopsied subjects with negative tTG-Ab were tested as control groups. IgA-AAA assays were performed by indirect immunofluorescence using rat epithelial intestinal cells, and by ELISA with a commercial kit. tTG-Ab assay was a radio-binding assay. Intestinal specimens were collected by upper endoscopy and the histological study was done according to the Marsh's classification modified by Oberhuber (M/O. Auto-antibodies assays and histological evaluation have been performed blindly by skilled operators. Results CD diagnosis was confirmed in 82 patients (type I M/O in 2 patients, IIIA in 18 patients, IIIB in 29 patients and IIIC in 33 patients. Two patients with type 1 lesion in presence of positive tTG-Ab and abdominal complaints, started a gluten free diet. The rate of IgA-AAA positivity (sensitivity by IFI and ELISA in histologically proven celiac disease patients, were 5.5% and 25% patients in IIIA, 27.5% and 34.4% patients in IIIB, 78.8% and 75% in IIIC patients, respectively. Patients with normal or nearly normal mucosa, regardless of tTG-Ab status, presented negative IgA-AAA IFI assay. On the other hand, 1 patient with normal mucosa but positive tTG-Ab, also presented positive IgA-AAA ELISA. All healthy non biopsied

  10. Indirect estimators in US federal programs

    CERN Document Server

    1996-01-01

    In 1991, a subcommittee of the Federal Committee on Statistical Methodology met to document the use of indirect estimators - that is, estimators which use data drawn from a domain or time different from the domain or time for which an estimate is required. This volume comprises the eight reports which describe the use of indirect estimators and they are based on case studies from a variety of federal programs. As a result, many researchers will find this book provides a valuable survey of how indirect estimators are used in practice and which addresses some of the pitfalls of these methods.

  11. A Novel High-Content Immunofluorescence Assay as a Tool to Identify at the Single Cell Level γ-Globin Inducing Compounds.

    Directory of Open Access Journals (Sweden)

    Marta Durlak

    Full Text Available The identification of drugs capable of reactivating γ-globin to ameliorate β-thalassemia and Sickle Cell anemia is still a challenge, as available γ-globin inducers still have limited clinical indications. High-throughput screenings (HTS aimed to identify new potentially therapeutic drugs require suitable first-step-screening methods combining the possibility to detect variation in the γ/β globin ratio with the robustness of a cell line. We took advantage of a K562 cell line variant expressing β-globin (β-K562 to set up a new multiplexed high-content immunofluorescence assay for the quantification of γ- and β-globin content at single-cell level. The assay was validated by using the known globin inducers hemin, hydroxyurea and butyric acid and further tested in a pilot screening that confirmed HDACs as targets for γ-globin induction (as proved by siRNA-mediated HDAC3 knockdown and by treatment with HDACs inhibitors entinostat and dacinostat and identified Heme-oxygenases as novel candidate targets for γ-globin induction. Indeed, Heme-oxygenase2 siRNA knockdown as well as its inhibition by Tin protoporphyrin-IX (TinPPIX greatly increased γ-globin expression. This result is particularly interesting as several metalloporphyrins have already been developed for clinical uses and could be tested (alone or in combination with other drugs to improve pharmacological γ-globin reactivation for the treatment of β-hemoglobinopathies.

  12. DNA Methylation Changes in Valproic Acid-Treated HeLa Cells as Assessed by Image Analysis, Immunofluorescence and Vibrational Microspectroscopy.

    Directory of Open Access Journals (Sweden)

    Giovana M B Veronezi

    Full Text Available Valproic acid (VPA, a well-known histone deacetylase inhibitor, has been reported to affect the DNA methylation status in addition to inducing histone hyperacetylation in several cell types. In HeLa cells, VPA promotes histone acetylation and chromatin remodeling. However, DNA demethylation was not checked in this cell model for standing effects longer than those provided by histone acetylation, which is a rapid and transient phenomenon. Demonstration of VPA-induced DNA demethylation in HeLa cells would contribute to understanding the effect of VPA on an aggressive tumor cell line. In the present work, DNA demethylation in VPA-treated HeLa cells was assessed by image analysis of chromatin texture, the abundance of 5-methylcytosine (5mC immunofluorescence signals and Fourier transform-infrared (FT-IR microspectroscopy centered on spectral regions related to the vibration of-CH3 groups. Image analysis indicated that increased chromatin unpacking promoted by a 4-h-treatment with 1.0 mM VPA persisted for 24 h in the absence of the drug, suggesting the occurrence of DNA demethylation that was confirmed by decreased 5mC immunofluorescence signals. FT-IR spectra of DNA samples from 1 mM or 20 mM VPA-treated cells subjected to a peak fitting analysis of the spectral window for-CH3 stretching vibrations showed decreased vibrations and energy of these groups as a function of the decreased abundance of 5mC induced by increased VPA concentrations. Only the 20 mM-VPA treatment caused an increase in the ratio of -CH3 bending vibrations evaluated at 1375 cm-1 in relation to in-plane vibrations of overall cytosines evaluated at 1492 cm-1. CH3 stretching vibrations showed to be more sensitive than-CH3 bending vibrations, as detected with FT-IR microspectroscopy, for studies aiming to associate vibrational spectroscopy and changes in DNA 5mC abundance.

  13. and Indigenous Indirect Enzyme-Linked Immunosorbent Assay

    Directory of Open Access Journals (Sweden)

    Annapurna S. Agasthya

    2012-01-01

    Full Text Available Brucellosis is one of the most important reemerging zoonoses in many countries. Brucellosis is caused by Gram-negative coccobacillus belonging to genus Brucella. Human brucellosis often makes the diagnosis difficult. The symptoms and clinical signs most commonly reported are fever, fatigue, malaise, chills, sweats headaches, myalgia, arthralgia, and weight loss. Some cases have been presented with only joint pain, lower backache, and involuntary limb movement, burning feet, or ischemic heart attacks. The focus of this work was to develop a highly sensitive and specific indirect ELISA by using smooth lipopolysaccharide antigen of Brucella abortus 99 to detect anti-Brucella antibodies at Project Directorate on Animal Disease Monitoring and Surveillance. Serum samples collected from 652 individuals in whom fever was not the major symptom but the complaint was of joint pain, headache, lower backache, and so forth, were screened by Rose Bengal plate agglutination test (RBPT and standard tube agglutination test (STAT. Subsequent testing of sera by indigenous indirect ELISA detected 20 samples positive (3.6% seroprevalence, and indirect ELISA was found to be more sensitive than RBPT and STAT. The seroprevalence in South Karnataka was 2.14%, and in North Karnataka it was 0.92%.

  14. Indirect Competitive Enzyme-Linked Immunosorbent Assay (ELISA).

    Science.gov (United States)

    Kohl, Thomas O; Ascoli, Carl A

    2017-07-05

    The indirect competitive ELISA (indirect cELISA) pits plate-immobilized antigen against antigens in solution for binding to antigen-specific antibody. The antigens in solution are in the test sample and are first incubated with antigen-specific antibody. These antibody-antigen complexes are then added to microtiter plates whose wells have been coated with purified antigen. The wells are washed to remove unbound antigen-antibody complexes and free antigen. A reporter-labeled secondary antibody is then added followed by the addition of substrate. Substrate hydrolysis yields a signal that is inversely proportional to antigen concentration within the sample. This is because when antigen concentration is high in the test sample, most of the antibody is bound before adding the solution to the plate. Most of the antibody remains in solution (as complexes) and is thus washed away before the addition of the reporter-labeled secondary antibody and substrate. Thus, the higher the antigen concentration in the test sample, the weaker the resultant signal in the detection step. The indirect cELISA is often used for competitive detection and quantification of antibodies against viral diseases in biological samples. © 2017 Cold Spring Harbor Laboratory Press.

  15. Isolation, Culture, Functional Assays, and Immunofluorescence of Myofiber-Associated Satellite Cells.

    Science.gov (United States)

    Vogler, Thomas O; Gadek, Katherine E; Cadwallader, Adam B; Elston, Tiffany L; Olwin, Bradley B

    2016-01-01

    Adult skeletal muscle stem cells, termed satellite cells, regenerate and repair the functional contractile cells in adult skeletal muscle called myofibers. Satellite cells reside in a niche between the basal lamina and sarcolemma of myofibers. Isolating single myofibers and their associated satellite cells provides a culture system that partially mimics the in vivo environment. We describe methods for isolating and culturing intact individual myofibers and their associated satellite cells from the mouse extensor digitorum longus muscle. Following dissection and isolation of individual myofibers we provide protocols for myofiber transplantation, satellite cell transfection, immune detection of satellite cell antigens, and assays to examine satellite cell self-renewal and proliferation.

  16. A direct view by immunofluorescent comet assay (IFCA) of DNA damage induced by nicking and cutting enzymes, ionizing (137)Cs radiation, UV-A laser microbeam irradiation and the radiomimetic drug bleomycin.

    Science.gov (United States)

    Grigaravicius, Paulius; Rapp, Alexander; Greulich, Karl Otto

    2009-03-01

    In DNA repair research, DNA damage is induced by different agents, depending on the technical facilities of the investigating researchers. A quantitative comparison of different investigations is therefore often difficult. By using a modified variant of the neutral comet assay, where the histone H1 is detected by immunofluorescence [immunofluorescent comet assay (IFCA)], we achieve previously unprecedented resolution in the detection of fragmented chromatin and show that trillions of ultraviolet A photons (of a few eV), billions of bleomycin (BLM) molecules and thousands of gamma quanta (of 662 keV) generate, in first order, similar damage in the chromatin of HeLa cells. A somewhat more detailed inspection shows that the damage caused by 20 Gy ionizing radiation and by a single laser pulse of 10 microJ are comparable, while the damage caused by 12 microg/ml BLM depends highly on the individual cell. Taken together, this work provides a detailed view of DNA fragmentation induced by different treatments and allows comparing them to some extent, especially with respect to the neutral comet assay.

  17. Optimization of Single- and Dual-Color Immunofluorescence Protocols for Formalin-Fixed, Paraffin-Embedded Archival Tissues.

    Science.gov (United States)

    Kajimura, Junko; Ito, Reiko; Manley, Nancy R; Hale, Laura P

    2016-02-01

    Performance of immunofluorescence staining on archival formalin-fixed paraffin-embedded human tissues is generally not considered to be feasible, primarily due to problems with tissue quality and autofluorescence. We report the development and application of procedures that allowed for the study of a unique archive of thymus tissues derived from autopsies of individuals exposed to atomic bomb radiation in Hiroshima, Japan in 1945. Multiple independent treatments were used to minimize autofluorescence and maximize fluorescent antibody signals. Treatments with NH3/EtOH and Sudan Black B were particularly useful in decreasing autofluorescent moieties present in the tissue. Deconvolution microscopy was used to further enhance the signal-to-noise ratios. Together, these techniques provide high-quality single- and dual-color fluorescent images with low background and high contrast from paraffin blocks of thymus tissue that were prepared up to 60 years ago. The resulting high-quality images allow the application of a variety of image analyses to thymus tissues that previously were not accessible. Whereas the procedures presented remain to be tested for other tissue types and archival conditions, the approach described may facilitate greater utilization of older paraffin block archives for modern immunofluorescence studies. © 2016 The Histochemical Society.

  18. Bioelectrochemical Systems for Indirect Biohydrogen Production

    KAUST Repository

    Regan, John M.; Yan, Hengjing

    2014-01-01

    by exoelectrogens at the anode. As an indirect approach to biohydrogen production, these systems are not subject to the hydrogen yield constraints of fermentative processes and have been proven to work with virtually any biodegradable organic substrate

  19. The WIMP Forest: Indirect Detection of a Chiral Square

    Energy Technology Data Exchange (ETDEWEB)

    Bertone, Gianfranco; Jackson, C.B.; Shaughnessy, Gabe; Tait, Tim M.P.; Vallinotto, Alberto

    2009-04-01

    The spectrum of photons arising from WIMP annihilation carries a detailed imprint of the structure of the dark sector. In particular, loop-level annihilations into a photon and another boson can in principle lead to a series of lines (a WIMP forest) at energies up to the WIMP mass. A specific model which illustrates this feature nicely is a theory of two universal extra dimensions compactified on a chiral square. Aside from the continuum emission, which is a generic prediction of most dark matter candidates, we find a 'forest' of prominent annihilation lines that, after convolution with the angular resolution of current experiments, leads to a distinctive (2-bump plus continuum) spectrum, which may be visible in the near future with the Fermi Gamma-Ray Space Telescope (formerly known as GLAST).

  20. Indirect techniques in nuclear astrophysics: a review

    International Nuclear Information System (INIS)

    Tribble, R E; Mukhamedzhanov, A M; Bertulani, C A; Cognata, M La; Spitaleri, C

    2014-01-01

    In this review, we discuss the present status of three indirect techniques that are used to determine reaction rates for stellar burning processes, asymptotic normalization coefficients, the Trojan Horse method and Coulomb dissociation. A comprehensive review of the theory behind each of these techniques is presented. This is followed by an overview of the experiments that have been carried out using these indirect approaches. (review article)

  1. A Comparison of Anti-Nuclear Antibody Quantification Using Automated Enzyme Immunoassays and Immunofluorescence Assays

    DEFF Research Database (Denmark)

    Baronaite, Renata; Engelhart, Merete; Mørk Hansen, Troels

    2014-01-01

    using IFA and automated EIA techniques. The IFA results generated by two independent laboratories were compared with the EIA results from antibodies against double-stranded DNA (dsDNA), from ANA screening, and from tests of the seven included subantigens. The final IFA and EIA results for 386 unique......, with Cohen's kappa value of 0.30 (95% confidence interval (CI) = 0.14-0.46), which decreased to 0.23 (95% CI = 0.06-0.40) when the results for dsDNA were omitted. The EIA method was less reliable for assessing nuclear and speckled reactivity patterns, whereas the IFA method presented difficulties detecting...... dsDNA and Ro activity. The automated EIA method was performed in a similar way to the conventional IFA method using HEp-2 cells; thus, automated EIA may be used as a screening test....

  2. Anti-dog IgG secondary antibody successfully detects IgG in a variety of aquatic mammals

    Science.gov (United States)

    Roehl, Katherine; Jankowski, Mark D.; Hofmeister, Erik K.

    2016-01-01

    Serological tests play an important role in the detection of wildlife diseases. However, while there are many commercial assays and reagents available for domestic species, there is a need to develop efficient serological assays for wildlife. In recent years, marine mammals have represented a wildlife group with emerging infectious diseases, such as influenza, brucellosis, and leptospirosis. However, with the exception of disease-agent-specific assays or functional assays, few reports describe the use of antibody detection assays in marine mammals. In an indirect enzyme-linked immunoassay (EIA) or an immunofluorescence assay, antibody is detected using an antitarget species secondary conjugated antibody. The sensitivity of the assay depends on the avidity of the binding reaction between the bound antibody and the detection antibody. A commercial polyclonal antidog IgG conjugated antibody was tested in an EIA for its ability to sensitively detect the IgG of seven marine mammals including sea otter (Enhydra lutris), polar bear (Ursus maritimus), grey seal (Halichoerus grypus), harbor seal (Phoca vitulina), northern elephant seal (Mirounga angustirostris), California sea lion (Zalophus californianus), Pacific walrus (Odobenus rosmarus) and one freshwater mammal: Asian small-clawed otter (Aonyx cinerea). With the exception of Asian small-clawed sea otters, the detection of IgG in these marine mammals either exceeded or was nearly equal to detection of dog IgG. The use of the tested commercial antidog IgG antibody may be a valid approach to the detection of antibody response to disease in sea mammals.

  3. An Indirect Route for Ethanol Production

    Energy Technology Data Exchange (ETDEWEB)

    Eggeman, T.; Verser, D.; Weber, E.

    2005-04-29

    The ZeaChem indirect method is a radically new approach to producing fuel ethanol from renewable resources. Sugar and syngas processing platforms are combined in a novel way that allows all fractions of biomass feedstocks (e.g. carbohydrates, lignins, etc.) to contribute their energy directly into the ethanol product via fermentation and hydrogen based chemical process technologies. The goals of this project were: (1) Collect engineering data necessary for scale-up of the indirect route for ethanol production, and (2) Produce process and economic models to guide the development effort. Both goals were successfully accomplished. The projected economics of the Base Case developed in this work are comparable to today's corn based ethanol technology. Sensitivity analysis shows that significant improvements in economics for the indirect route would result if a biomass feedstock rather that starch hydrolyzate were used as the carbohydrate source. The energy ratio, defined as the ratio of green energy produced divided by the amount of fossil energy consumed, is projected to be 3.11 to 12.32 for the indirect route depending upon the details of implementation. Conventional technology has an energy ratio of 1.34, thus the indirect route will have a significant environmental advantage over today's technology. Energy savings of 7.48 trillion Btu/yr will result when 100 MMgal/yr (neat) of ethanol capacity via the indirect route is placed on-line by the year 2010.

  4. Flow Cytometry Detection of Infectious Rotaviruses in Environmental and Clinical Samples

    Science.gov (United States)

    Abad, F. Xavier; Pintó, Rosa M.; Bosch, Albert

    1998-01-01

    A method for the detection of infectious human rotaviruses based on infection of CaCo-2 cells and detection of infected cells by indirect immunofluorescence and flow cytometry (IIF-FC) has been developed. The technique was validated by performing a seminested reverse transcription-PCR assay with sorted cell populations. The efficiency of the procedure has been compared with that of the standard method of infection of MA104 cells and ulterior detection by IIF and optical microscopy (IIF-OM) and with that of infection of MA104 cells and detection by IIF-FC. The limit of sensitivity for the detection of the cell-adapted strain Itor P13, expressed as the most probable number of cytopathogenic units, was established as 200 and 2 for MA104 and CaCo-2 cells, respectively, by the IIF-FC method. The ratio of infectious virus particles to total virus particles for a wild-type rotavirus was determined to be 1/2 × 106 and 1/2 × 104 for IIF-OM with MA104 cells and IIF-FC with CaCo-2 cells, respectively. The use of IIF-FC with CaCo-2 cells was tested with fecal and water samples and proved to be more effective than the standard procedure for rotavirus detection. PMID:9647805

  5. Imunofluorescência utilizando isolados brasileiros no diagnóstico sorológico de infecção por lentivírus em caprinos Immunofluorescence using Brazilian isolates for serological diagnosis of lentivirus infection in goats

    Directory of Open Access Journals (Sweden)

    Dilmara Reischak

    2002-01-01

    immunofluorescent antibody (IFA test using Brazilian SRLV isolates for serological diagnosis of infection in goats. The IFA test was compared in its sensitivity and specificity to the AGID test in which Maedi-Visna WLC-1 strain was used as antigen. Ovine synovial secondary cell cultures infected with two goat SRLV isolates (CAEV UFRGS/Br-2 and CAEV UFRGS/Br-5 were used for the IFA. Goat serum samples (n = 239 were tested by the two tests. The AGID test detected antibodies in 129 (53.9% serum samples. A higher number of animals was considered positive for the IFA. However, different results were obtained depending on the isolate used for the antigen preparation. When CAEV UFRGS/Br-2 was used as antigen, 216 (90.3% serum samples tested positive, against 213 (89.1% with CAEV UFRGS/Br-5. There was no significant statistical difference between the antigens prepared with these two isolates. The IFA had sensitivity of 94.6% and 96.9%, and specificity of 14.5 % and 20%, when CAEV UFRGS/Br-2 and CAEV UFRGS/Br-5 were used as antigens, respectively. These results demonstrate that the IFA with antigens prepared with Brazilian SRLV isolates may play an important a role as a complementary serological test for the diagnosis of infections by these agents.

  6. Dual-color immunofluorescent labeling with quantum dots of the diabetes-associated proteins aldose reductase and Toll-like receptor 4 in the kidneys of diabetic rats

    Directory of Open Access Journals (Sweden)

    Liu XM

    2015-05-01

    sensitivity and specificity for the detection of biomarkers in cells and tissues, but also is a valuable supplement of immunohistochemistry. The QD-based multiplexed imaging technology provides a new insight into the mechanistic study of the correlation among biological factors as well as having potential applications in the diagnosis and treatment of diseases. Keywords: diabetic nephropathy, TLR4, immunofluorescence, immunohistochemistry

  7. Development and application of radioimmunoassays and enzyme immunoassays in microbiological and immunological diagnosis. 2. Comparative studies for the detection of toxoplasma antibodies with ELISA, RIA and other serological methods

    Energy Technology Data Exchange (ETDEWEB)

    Mueller, W A; Struy, H; Holzwarth, F [Medizinische Akademie, Magdeburg (German Democratic Republic)

    1982-06-01

    Comparative studies of indirect immunofluorescence test (IT), complement binding reaction (CBR), enzyme- and radioimmunoassay (ELISA, RIA) for the detection of toxoplasma antibodies in sera of 513 patients are reported. The precision dependent on time, showed coefficients of variation from 3% to 12% (IFT 3%, CBR 10%, ELISA 12%, RIA 7%). The correlation of IFT and ELISA as well as RIA was relatively unfavourable (coefficient of correlation IFT/ELISA r = 0.52, IFT/RIA r = 0.54, RIA/ELISA r = 0.60). The ELISA is the most sensitive method for the detection of antibodies. The specificity of the Toxo-ELISA has to be improved by application of suitable fractions of antigens.

  8. Analysis of indirect taxation in Ukraine

    Directory of Open Access Journals (Sweden)

    Khodyakova Olga V.

    2014-01-01

    Full Text Available The goal of the article is analysis of the structure and dynamics of indirect taxes for the previous five years and also the influence of indirect taxation upon formation of income of the State Budget of Ukraine. The article analyses the modern state of indirect taxation in Ukraine. Specific weight of the value added tax, excise tax and customs duty are considered in the structure of tax receipts of the consolidated budget of Ukraine as indirect taxes. The article shows that receipts of the State Budget of Ukraine are mostly provided by indirect taxes. The Ukrainian taxation system is mostly a factor of reduction of the level of economic growth and investment activity in the country and the existing system of administering is not completely capable of excluding the possibility of tax evasion. The prospect of further studies in this direction is improvement of organisation of tax control in Ukraine and differentiation of the value added tax rates depending on the level of consumption of goods and level of income of consumers.

  9. Comparison of viral RNA electrophoresis and indirect ELISA methods in the diagnosis of human rotavirus infection

    Energy Technology Data Exchange (ETDEWEB)

    Avendano, L F; Dubinovsky, S; James, Jr, H D

    1984-01-01

    A total of 177 stool samples from Chilean diarrhea patients under two years of age were tested for rotavirus by two methods - the indirect enzyme-linked immunosorbent assay (indirect ELISA) and viral RNA electrophoresis in agarose gels (v RNA EPH). Fifty of the specimens came from patients with acute diarrhea and 127 came from patients with protracted diarrhea. The indirect ELISA testing was performed at the National Institutes of Health in the United States: the electrophoretic testing was carried out in Santiago, Chile by the authors. The electrophoretic method detected rotavirus in 36% of the acute samples and 25% of the samples from protracted cases, while the indirect ELISA method detected rotavirus in higher percentages of samples - 46% and 38%, respectively. These results support the conclusion that v RNA EPH is a less sensitive method for detecting rotavirus than the indirect ELISA. Nevertheless, the former method's high specificity, ease of application, and low cost make it a worthwhile alternative to indirect ELISA. Thus, considering the important role played by rotavirus in infant diarrhea and the need for a diagnostic technique that can be incorporated into the routines of medical center laboratories in developing countries, there is good reason to conclude that v RNA EPH is a useful tool for studying rotavirus diarrhea. 18 refs, 3 tabs. Also published in the Bol. Oficina Sanit. Panam. (1984) v. 97(1), p. 1-7 (In Spanish).

  10. Comparison of viral RNA electrophoresis and indirect ELISA methods in the diagnosis of human rotavirus infection

    International Nuclear Information System (INIS)

    Avendano, L.F.; Dubinovsky, S.

    1984-01-01

    A total of 177 stool samples from Chilean diarrhea patients under two years of age were tested for rotavirus by two methods - the indirect enzyme-linked immunosorbent assay (indirect ELISA) and viral RNA electrophoresis in agarose gels (v RNA EPH). Fifty of the specimens came from patients with acute diarrhea and 127 came from patients with protracted diarrhea. The indirect ELISA testing was performed at the National Institutes of Health in the United States: the electrophoretic testing was carried out in Santiago, Chile by the authors. The electrophoretic method detected rotavirus in 36% of the acute samples and 25% of the samples from protracted cases, while the indirect ELISA method detected rotavirus in higher percentages of samples - 46% and 38%, respectively. These results support the conclusion that v RNA EPH is a less sensitive method for detecting rotavirus than the indirect ELISA. Nevertheless, the former method's high specificity, ease of application, and low cost make it a worthwhile alternative to indirect ELISA. Thus, considering the important role played by rotavirus in infant diarrhea and the need for a diagnostic technique that can be incorporated into the routines of medical center laboratories in developing countries, there is good reason to conclude that v RNA EPH is a useful tool for studying rotavirus diarrhea. (author)

  11. Relevance of indirect transmission for wildlife disease surveillance

    Directory of Open Access Journals (Sweden)

    Martin Lange

    2016-11-01

    Full Text Available Epidemiological models of infectious diseases are essential tools in support of risk assessment, surveillance design and contingency planning in public and animal health. Direct pathogen transmission from host to host is an essential process of each host-pathogen system and respective epidemiological modelling concepts. It is widely accepted that numerous diseases involve indirect transmission through pathogens shed by infectious hosts to their environment. However, epidemiological models largely do not represent pathogen persistence outside the host explicitly. We hypothesize that this simplification might bias management-related model predictions for disease agents that can persist outside their host for a certain time span. We adapted an individual-based, spatially explicit epidemiological model that can mimic both transmission processes. One version explicitly simulated indirect pathogen transmission through a contaminated environment. A second version simulated direct host-to-host transmission only. We aligned the model variants by the transmission potential per infectious host (i.e. basic reproductive number R0 and the spatial transmission kernel of the infection to allow unbiased comparison of predictions. The quantitative model results are provided for the example of surveillance plans for early detection of foot-and-mouth disease in wild boar, a social host.We applied systematic sampling strategies on the serological status of randomly selected host individuals in both models. We compared between the model variants the time to detection and the area affected prior to detection, measures that strongly influence mitigation costs. Moreover, the ideal sampling strategy to detect the infection in a given time frame was compared between both models.We found the simplified, direct transmission model to underestimate necessary sample size by up to one order of magnitude, but to overestimate the area put under control measures. Thus, the model

  12. A comparative investigation of seven indirect attitude measures.

    Science.gov (United States)

    Bar-Anan, Yoav; Nosek, Brian A

    2014-09-01

    We compared the psychometric qualities of seven indirect attitude measures across three attitude domains (race, politics, and self-esteem) with a large sample (N = 23,413). We compared the measures on internal consistency, sensitivity to known effects, relationships with indirect and direct measures of the same topic, the reliability and validity of single-category attitude measurement, their ability to detect meaningful variance among people with nonextreme attitudes, and their robustness to the exclusion of misbehaving or well-behaving participants. All seven indirect measures correlated with each other and with direct measures of the same topic. These relations were always weak for self-esteem, moderate for race, and strong for politics. This pattern suggests that some of the sources of variation in the reliability and predictive validity of the indirect measures is a function of the concepts rather than the methods. The Implicit Association Test (IAT) and Brief IAT (BIAT) showed the best overall psychometric quality, followed by the Go–No-Go association task, Single-Target IAT (ST-IAT), Affective Misattribution Procedure (AMP), Sorting Paired Features task, and Evaluative Priming. The AMP showed a steep decline in its psychometric qualities when people with extreme attitude scores were removed. Single-category attitude scores computed for the IAT and BIAT showed good relationships with other attitude measures but no evidence of discriminant validity between paired categories. The other measures, especially the AMP and ST-IAT, showed better evidence for discriminant validity. These results inform us on the validity of the measures as attitude assessments, but do not speak to the implicitness of the measured constructs.

  13. Accuracy of five intraoral scanners compared to indirect digitalization.

    Science.gov (United States)

    Güth, Jan-Frederik; Runkel, Cornelius; Beuer, Florian; Stimmelmayr, Michael; Edelhoff, Daniel; Keul, Christine

    2017-06-01

    Direct and indirect digitalization offer two options for computer-aided design (CAD)/ computer-aided manufacturing (CAM)-generated restorations. The aim of this study was to evaluate the accuracy of different intraoral scanners and compare them to the process of indirect digitalization. A titanium testing model was directly digitized 12 times with each intraoral scanner: (1) CS 3500 (CS), (2) Zfx Intrascan (ZFX), (3) CEREC AC Bluecam (BLU), (4) CEREC AC Omnicam (OC) and (5) True Definition (TD). As control, 12 polyether impressions were taken and the referring plaster casts were digitized indirectly with the D-810 laboratory scanner (CON). The accuracy (trueness/precision) of the datasets was evaluated by an analysing software (Geomagic Qualify 12.1) using a "best fit alignment" of the datasets with a highly accurate reference dataset of the testing model, received from industrial computed tomography. Direct digitalization using the TD showed the significant highest overall "trueness", followed by CS. Both performed better than CON. BLU, ZFX and OC showed higher differences from the reference dataset than CON. Regarding the overall "precision", the CS 3500 intraoral scanner and the True Definition showed the best performance. CON, BLU and OC resulted in significantly higher precision than ZFX did. Within the limitations of this in vitro study, the accuracy of the ascertained datasets was dependent on the scanning system. The direct digitalization was not superior to indirect digitalization for all tested systems. Regarding the accuracy, all tested intraoral scanning technologies seem to be able to reproduce a single quadrant within clinical acceptable accuracy. However, differences were detected between the tested systems.

  14. An experimental test for indirect benefits in Drosophila melanogaster

    Directory of Open Access Journals (Sweden)

    Ödeen Anders

    2007-03-01

    Full Text Available Abstract Background Despite much empirical attention, tests for indirect benefits of mate choice have rarely considered the major components of sexual and nonsexual offspring fitness relevant to a population. Here we use a novel experimental design to test for the existence of any indirect benefits in a laboratory adapted population of D. melanogaster. Our experiment compared the fitness (mating success, longevity, and productivity of individuals possessing genomes that derived two generations previously from males that were either entirely successful (studs or wholly unsuccessful (duds at achieving mates in three subsequent rounds of mating trials. Results Males from the stud treatment were 30% more successful on average at securing mates than males from the dud treatment. In contrast, we found no difference between treatments in measures of productivity or of longevity when measured in a mixed-sex environment. In the absence of females, however, males in the stud treatment outlived males in the dud treatment. Conclusion Our results suggest that mating with successful males in this population provides an indirect benefit to females and that, at least in this environment, the benefit arises primarily through the production of more attractive male offspring. However, it is unclear whether this represents solely a traditional sexy sons benefit or whether there is an additional good genes component (with male offspring simply allocating their surplus condition to traits that enhance their mating success. The lack of any detectable differences in female fitness between the two treatments suggests the former, although the longevity advantage of males in the stud treatment when females were absent is consistent with the latter. Determining the effect of this indirect benefit on the evolution of female mate preferences (or resistance will require comparable data on the direct costs of mating with various males, and an understanding of how these costs

  15. A review of Indirect Matrix Converter Topologies

    Directory of Open Access Journals (Sweden)

    Salem Rahmani

    2015-08-01

    Full Text Available Abstract—Matrix Converter (MC is a modern direct AC/AC electrical power converter without dc-link capacitor. MC is operated in four quadrant, assuring a control of the output voltage, amplitude and frequency. The matrix converter has recently attracted significant attention among researchers and it has become increasing attractive for applications of wind energy conversion, military power supplies, induction motor drives, etc. Recently, different MC topologies have been proposed and developed which have their own advantages and disadvantages. Matrix converter can be classified as direct and indirect structures. The direct one has been elaborated in previous work. In this paper the indirect MCs are reviewed. Different characteristics of the indirect MC topologies are mentioned to show the strengths and weaknesses of such converter topologies.

  16. Indirect taxes on food in Southern Brazil

    Directory of Open Access Journals (Sweden)

    Denize Mirian da Silva

    2013-12-01

    Full Text Available The objective of this paper is to estimate the indirect tax burden on food for ten income classes, based on income and household total expenditure in southern Brazil. Thus it can be seen as indirect taxes on foods affect the monetary income and consumption pattern of households. To reach the objectives proposed, will be used the Pintos-Payeras (2008 model. The database iscomposed by microdata from the Household Budgeting Survey (POF 2008-2009 and the tax regulations of the country and the southern states of Brazil. The results show that indirect taxes on food in Southern Brazil is regressive when based on income and expenditure of household , ie , the poorest people pay proportionately more taxes and have their consumption pattern highest taxed ICMS (Brazilian value added tax is the tax that contributes most to the regressivity.

  17. Ballistic fractures: indirect fracture to bone.

    Science.gov (United States)

    Dougherty, Paul J; Sherman, Don; Dau, Nathan; Bir, Cynthia

    2011-11-01

    Two mechanisms of injury, the temporary cavity and the sonic wave, have been proposed to produce indirect fractures as a projectile passes nearby in tissue. The purpose of this study is to evaluate the temporal relationship of pressure waves using strain gauge technology and high-speed video to elucidate whether the sonic wave, the temporary cavity, or both are responsible for the formation of indirect fractures. Twenty-eight fresh frozen cadaveric diaphyseal tibia (2) and femurs (26) were implanted into ordnance gelatin blocks. Shots were fired using 9- and 5.56-mm bullets traversing through the gelatin only, passing close to the edge of the bone, but not touching, to produce an indirect fracture. High-speed video of the impact event was collected at 20,000 frames/s. Acquisition of the strain data were synchronized with the video at 20,000 Hz. The exact time of fracture was determined by analyzing and comparing the strain gauge output and video. Twenty-eight shots were fired, 2 with 9-mm bullets and 26 with 5.56-mm bullets. Eight indirect fractures that occurred were of a simple (oblique or wedge) pattern. Comparison of the average distance of the projectile from the bone was 9.68 mm (range, 3-20 mm) for fractured specimens and 15.15 mm (range, 7-28 mm) for nonfractured specimens (Student's t test, p = 0.036). In this study, indirect fractures were produced after passage of the projectile. Thus, the temporary cavity, not the sonic wave, was responsible for the indirect fractures.

  18. Indirect Comorbidity in Childhood and Adolescence

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    William eCopeland

    2013-11-01

    Full Text Available Objective: Comorbidity between psychiatric disorders is common, but pairwise associations between two disorders may be explained by the presence of other diagnoses that are associated with both disorders or indirect comorbidity. Method: Comorbidities of common childhood psychiatric disorders were tested in three community samples of children ages 6 to 17 (8931 observations of 2965 subjects. Psychiatric disorder status in all three samples was assessed with the Child and Adolescent Psychiatric Assessment. Indirect comorbidity was defined as A-B associations that decreased from significance to nonsignificance after adjusting for other disorders. Results: All tested childhood psychiatric disorders were positively associated in bivariate analyses. After adjusting for comorbidities, many ssociations involving a behavioral disorder and an emotional disorder were attenuated suggesting indirect comorbidity. Generalized anxiety and depressive disorders displayed a very high level of overlap (adjusted OR=37.9. All analyses were rerun with depressive disorders grouped with generalized anxiety disorder in a single distress disorders category. In these revised models, all associations between and emotional disorder and a behavior disorder met our criteria for indirect comorbidity except for the association of oppositional defiant disorder with distress disorders (OR=11.3. Follow-up analyses suggested that the indirect associations were primarily accounted for by oppositional defiant disorder and the distress disorder category. There was little evidence of either sex differences or differences by developmental period Conclusions: After accounting for the overlap between depressive disorders with generalized anxiety disorder, direct comorbidity between emotional and behavioral disorders was uncommon. When there was evidence of indirect comorbidity, ODD and distress disorders were the key intermediary diagnoses accounting for the apparent associations.

  19. Indirect effects in dual radiation action

    International Nuclear Information System (INIS)

    Zaider, M.; Rossi, H.H.

    1988-01-01

    The basic aim in this paper is to establish the link between indirect effects of radiation action and the spatial distribution of radicals at the time of energy deposition as well as throughout subsequent diffusion and interaction. The fact that radicals diffuse for a finite distance before damaging a biomolecule has dramatic effects on their subsequent probability to result in lesions. Thus at very low DMSO concentrations, where p = 0.5, one expects - all other things being equal - some 75% of the lesions to result from indirect or semidirect lesions. The number calculated here is lower (15%), a direct result of the fact that such lesions involve proximity functions modulated by diffusion. At higher DMSO concentrations this percentage becomes progressively smaller, as expected. It appears thus that for low-LET radiation, the relative amount of indirect damage in single tracks (also termed intratrack or single events) action is very small. By contrast, intertrack (or two-event) contributions will have the ratio between direct and (indirect + semidirect) contributions given by p 2 /(1-p 2 ). The reason for this is that sublesions from different tracks are uniformly distributed throughout the cell nucleus; their probability of interaction should not depend on any previous diffusional processes. For the example given above (p = 0.5) they do expect 65% of intertrack (two-hit) lesions to have resulted from indirect or semidirect mechanisms. This contrast between the almost exclusively direct character of intratrack lesions and the dominant role of indirect action in intertrack lesions produced by low-LET radiation is an important conclusion of this study

  20. The indirect radioiodination of vasoactive intestinal peptide

    International Nuclear Information System (INIS)

    Wang Lihua; Li Junling; Yin Duanzhi; Zhang Lei; Zhang Xiuli; Wang Yongxian

    2002-01-01

    Objective: To seek for an effective way to acquire radiolabeled vasoactive intestinal peptide (VIP) with excellent in vivo stability. N-succinimidyl-3-iodo-125-benzoate (S 125 IB) came from radioiodination of N-succinimidyl-3-(tri-n-butylstannyl) benzoate (ATE) precursor and then conjugated with VIP to form 125 IBA-VIP. The labelling procedure was optimized; the in vitro stability and biological activity were evaluated. Methods: 1) Radiolabeling of ATE precursor was achieved with iodogen oxidant and the influential factors were considered in this procedure. The labeling efficiency was determined by thin layer chromatography (TLC) and the purification was carried out by Sep-pak silica gel cartridge. The stability was detected by TLC after 2 h storage in dark at 4 degree C. 2) Conjugation of S 125 IB and VIP. The labelling efficiency was determined with RP TLC and the purification was carried out with high performance liquid chromatography (HPLC, RP C18 column). Trichloroacetic acid (TCA) precipitation method was applied to evaluate the in vitro stability while the biological activity was determined by cell binding experiments with SGC7901 cell lines. Results: 1) S 125 IB experiments. The radioiodination of ATE was performed well for 5 min at 25 degree C with 10 micrograms of iodogen at suitable mole ratio (3-8:1) of ATE/Na 125 I, the labelling efficiency was about 96%. The stability was kept well at 4 degree C in dark, no significant decrease of S 125 IB was observed. 2) The conjugation efficiency of S 125 IB and VIP was above 75% with TLC. HPLC showed the different retention time (t R ) as follows, 125 IBA-VIP: 13.3 min, S 125 IB: 19.6 min, VIP: 8.32 min. The stability of 125 IBA-VIP was better than 125 I-VIP from direct radioiodination of VIP with iodogen oxidant, only 2.85% decrease was found after 7 d at 4 degree C. The biological activity of 125 IBA-VIP was kept as well as 125 I-VIP under the condition of 37 degree C 60 min. Conclusions: The indirect

  1. Global indirect aerosol effects: a review

    Directory of Open Access Journals (Sweden)

    U. Lohmann

    2005-01-01

    Full Text Available Aerosols affect the climate system by changing cloud characteristics in many ways. They act as cloud condensation and ice nuclei, they may inhibit freezing and they could have an influence on the hydrological cycle. While the cloud albedo enhancement (Twomey effect of warm clouds received most attention so far and traditionally is the only indirect aerosol forcing considered in transient climate simulations, here we discuss the multitude of effects. Different approaches how the climatic implications of these aerosol effects can be estimated globally as well as improvements that are needed in global climate models in order to better represent indirect aerosol effects are discussed in this paper.

  2. Biomass Indirect Liquefaction Strategy Workshop Summary Report

    Energy Technology Data Exchange (ETDEWEB)

    none,

    2014-07-01

    This report is based on the proceedings of the U.S. Department of Energy Bioenergy Technologies Office Biomass Indirect Liquefaction Strategy Workshop. The workshop, held March 20–21, 2014, in Golden, Colorado, discussed and detailed the research and development needs for biomass indirect liquefaction. Discussions focused on pathways that convert biomass-based syngas (or any carbon monoxide, hydrogen gaseous stream) to liquid intermediates (alcohols or acids) and further synthesize those intermediates to liquid hydrocarbons that are compatible as either a refinery feed or neat fuel.

  3. The power of indirect social ties

    OpenAIRE

    Zuo, Xiang; Blackburn, Jeremy; Kourtellis, Nicolas; Skvoretz, John; Iamnitchi, Adriana

    2014-01-01

    While direct social ties have been intensely studied in the context of computer-mediated social networks, indirect ties (e.g., friends of friends) have seen little attention. Yet in real life, we often rely on friends of our friends for recommendations (of good doctors, good schools, or good babysitters), for introduction to a new job opportunity, and for many other occasional needs. In this work we attempt to 1) quantify the strength of indirect social ties, 2) validate it, and 3) empiricall...

  4. Indirect methods for wake potential integration

    International Nuclear Information System (INIS)

    Zagorodnov, I.

    2006-05-01

    The development of the modern accelerator and free-electron laser projects requires to consider wake fields of very short bunches in arbitrary three dimensional structures. To obtain the wake numerically by direct integration is difficult, since it takes a long time for the scattered fields to catch up to the bunch. On the other hand no general algorithm for indirect wake field integration is available in the literature so far. In this paper we review the know indirect methods to compute wake potentials in rotationally symmetric and cavity-like three dimensional structures. For arbitrary three dimensional geometries we introduce several new techniques and test them numerically. (Orig.)

  5. A sensitive, reproducible and objective immunofluorescence analysis method of dystrophin in individual fibers in samples from patients with duchenne muscular dystrophy.

    Directory of Open Access Journals (Sweden)

    Chantal Beekman

    Full Text Available Duchenne muscular dystrophy (DMD is characterized by the absence or reduced levels of dystrophin expression on the inner surface of the sarcolemmal membrane of muscle fibers. Clinical development of therapeutic approaches aiming to increase dystrophin levels requires sensitive and reproducible measurement of differences in dystrophin expression in muscle biopsies of treated patients with DMD. This, however, poses a technical challenge due to intra- and inter-donor variance in the occurrence of revertant fibers and low trace dystrophin expression throughout the biopsies. We have developed an immunofluorescence and semi-automated image analysis method that measures the sarcolemmal dystrophin intensity per individual fiber for the entire fiber population in a muscle biopsy. Cross-sections of muscle co-stained for dystrophin and spectrin have been imaged by confocal microscopy, and image analysis was performed using Definiens software. Dystrophin intensity has been measured in the sarcolemmal mask of spectrin for each individual muscle fiber and multiple membrane intensity parameters (mean, maximum, quantiles per fiber were calculated. A histogram can depict the distribution of dystrophin intensities for the fiber population in the biopsy. This method was tested by measuring dystrophin in DMD, Becker muscular dystrophy, and healthy muscle samples. Analysis of duplicate or quadruplicate sections of DMD biopsies on the same or multiple days, by different operators, or using different antibodies, was shown to be objective and reproducible (inter-assay precision, CV 2-17% and intra-assay precision, CV 2-10%. Moreover, the method was sufficiently sensitive to detect consistently small differences in dystrophin between two biopsies from a patient with DMD before and after treatment with an investigational compound.

  6. Anti-Trypanosoma cruzi antibody detection in eastern Andalusia (Spain).

    Science.gov (United States)

    Marín, Clotilde; Concha-Valdez, Fanny; Cañas, Rocío; Gutiérrez-Sánchez, Ramón; Sánchez-Moreno, Manuel

    2014-03-01

    Chagas disease caused by the protozoan haemoflagellate Trypanosoma cruzi is no longer found exclusively in Latin America; the disease is occurring in Europe, and Spain is the country with the highest prevalence. Our aim was to detect anti-T. cruzi antibodies in blood donors from southeast Spain, and we performed eight serological diagnostic assays on each of 550 blood samples collected in March-June 2010. Two in-house ELISA methods were used to test against a parasite lysate (ELISA-H) and the semi-purified superoxide dismutase excreted by T. cruzi (ELISA-SODe); we also used the Western blot technique against the same antigen (WB-SODe), indirect immunofluorescence (IFA) and four commercial tests. The serological test results showed a range of seroprevalence values, the lowest being 1.1%, determined by IFA and two commercial tests (Ab rapid and Chagascreen); other values were: 1.3% (commercial ELISA [Chagas ELISA IgG+IgM]); 2.1% (immunochromatographic test [Stick Chagas]); 2.7% (ELISA-H); 4.0% (WB-SODe); and 4.2%, the highest value (ELISA-SODe). The excellent specificity of SODe antigen for the detection of antibodies to T. cruzi in donors lead us to affirm that the serological test performed with this biomarker could provide a useful screening and confirmatory test method for cases of Chagas disease.

  7. Development of indirect sandwich ELISA for determination of excretory-secretory antigens of Fasciola hepatica

    Directory of Open Access Journals (Sweden)

    Libertad Alzamora-Gonzales

    2016-05-01

    Full Text Available Fasciolosis is a cosmopolitan parasitosis medical-veterinary importance caused by Fasciola hepatica, which affects sheep, goats and cattle; and it affects man accidentally causing an epidemic-endemic infection difficult to diagnose. The aim was to develop an indirect sandwich ELISA with 3 antibodies for detecting excretory-secretory antigens of Fasciola hepatica (ESFh. For the development of indirect sandwich ELISA were used, as capture antibody, mouse polyclonal antibodies anti ESFh and polyclonal antibodies rabbit anti-ESFh as detection antibody, at the concentrations of 10 and 5 µg/mL respectively. The conjugate used was mouse monoclonal anti- total immunoglobulins rabbit linked to peroxidase (1/1000. Were analized 31 sheep fecal samples, and the results were compared with those obtained by direct coproparasitological examination (DC and counterimmunoelectrophoresis (CIEP. The detection limit obtained for indirect sandwich ELISA was 100 ng/mL. The test had a 100% sensitivity, 96.6% specificity, positive and negative predictive values of 50% and 96.6% respectively, in relation to DC test. Comparing with CIEP the specificity obtained for indirect sandwich ELISA was 93.5% and a negative predictive value of 100%. We concluded that indirect sandwich ELISA designed is able to detect metabolic antigens in ovine feces samples and can be used for Fasciola hepatica diagnosis.

  8. Indirect processes in electron-ion scattering

    International Nuclear Information System (INIS)

    Bottcher, C.; Griffin, D.C.; Pindzola, M.S.; Phaneuf, R.A.

    1983-10-01

    A summary is given of an informal workshop held at Oak Ridge National Laboratory on June 22-23, 1983, in which the current status of theoretical calculations of indirect processes in electron-ion scattering was reviewed. Processes of particular interest in astrophysical and fusion plasmas were emphasized. Topics discussed include atomic structure effects, electron-impact ionization, and dielectronic recombination

  9. Indirect effects of recreation on wildlife

    Science.gov (United States)

    David N. Cole; Peter B. Landres

    1995-01-01

    Most of this book focuses on direct impacts to wildlife that result from contact with people. The purpose of our chapter is to provide a broad overview of the indirect influences that recreation has on wildlife. Recreational activities can change the habitat of an animal. This, in turn, affects the behavior, survival, reproduction, and distribution of individuals....

  10. 46 CFR 154.1720 - Indirect refrigeration.

    Science.gov (United States)

    2010-10-01

    ... Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CERTAIN BULK DANGEROUS CARGOES SAFETY STANDARDS FOR SELF-PROPELLED VESSELS CARRYING BULK LIQUEFIED GASES Special Design and Operating Requirements § 154.1720 Indirect refrigeration. A refrigeration system that is used to cool acetaldehyde, ethylene...

  11. Indirect processes in electron-ion scattering

    Energy Technology Data Exchange (ETDEWEB)

    Bottcher, C.; Griffin, D.C.; Pindzola, M.S.; Phaneuf, R.A.

    1983-10-01

    A summary is given of an informal workshop held at Oak Ridge National Laboratory on June 22-23, 1983, in which the current status of theoretical calculations of indirect processes in electron-ion scattering was reviewed. Processes of particular interest in astrophysical and fusion plasmas were emphasized. Topics discussed include atomic structure effects, electron-impact ionization, and dielectronic recombination.

  12. 27 CFR 6.32 - Indirect interest.

    Science.gov (United States)

    2010-04-01

    ... OF THE TREASURY LIQUORS âTIED-HOUSEâ Unlawful Inducements Interest in Retail Property § 6.32 Indirect interest. Industry member interest in retail property includes any interest acquired by corporate officials, partners, employees or other representatives of the industry member. Any interest in retail property...

  13. 27 CFR 6.26 - Indirect interest.

    Science.gov (United States)

    2010-04-01

    ... OF THE TREASURY LIQUORS âTIED-HOUSEâ Unlawful Inducements Interest in Retail License § 6.26 Indirect interest. Industry member interest in retail licenses includes any interest acquired by corporate officials, partners, employees or other representatives of the industry member. Any interest in a retail license...

  14. Indirect Costs: Daily Bread, Cake, or Cracker.

    Science.gov (United States)

    Lucas, Robert A.

    1988-01-01

    The tradition of negotiating indirect costs in grants should be abandoned, and research administrators should instead offer different levels of service depending on what the sponsor wants to spend. Three levels of overhead rate are suggested (super, regular, and economy) and their corresponding levels of service are defined. (MSE)

  15. Epidemic spreading through direct and indirect interactions.

    Science.gov (United States)

    Ganguly, Niloy; Krueger, Tyll; Mukherjee, Animesh; Saha, Sudipta

    2014-09-01

    In this paper we study the susceptible-infected-susceptible epidemic dynamics, considering a specialized setting where popular places (termed passive entities) are visited by agents (termed active entities). We consider two types of spreading dynamics: direct spreading, where the active entities infect each other while visiting the passive entities, and indirect spreading, where the passive entities act as carriers and the infection is spread via them. We investigate in particular the effect of selection strategy, i.e., the way passive entities are chosen, in the spread of epidemics. We introduce a mathematical framework to study the effect of an arbitrary selection strategy and derive formulas for prevalence, extinction probabilities, and epidemic thresholds for both indirect and direct spreading. We also obtain a very simple relationship between the extinction probability and the prevalence. We pay special attention to preferential selection and derive exact formulas. The analysis reveals that an increase in the diversity in the selection process lowers the epidemic thresholds. Comparing the direct and indirect spreading, we identify regions in the parameter space where the prevalence of the indirect spreading is higher than the direct one.

  16. Indirect Determination of the sigma - omega

    DEFF Research Database (Denmark)

    Ulfkjær, J. P.; Brincker, Rune

    . The beams where submitted to three-point bending in a servo-controlled materials testing system. The constitutive parameters in the fictitious crack model were determined by using three different indirect techniques, which are described and evaluated. The results show that the fracture energy is increasing...

  17. Comprehension Process of Second Language Indirect Requests.

    Science.gov (United States)

    Takahashi, Satomi; Roitblat, Herbert L.

    1994-01-01

    Examines the comprehension of English conventional indirect requests by native English speakers and Japanese learners of English. Subjects read stories inducing either a conventional or a literal interpretation of a priming sentence. Results suggest that both native and nonnative speakers process both meanings of an ambiguous conventional request.…

  18. 48 CFR 31.203 - Indirect costs.

    Science.gov (United States)

    2010-10-01

    ... fiscal year used for financial reporting purposes in accordance with generally accepted accounting... contract or other work, indirect costs are those remaining to be allocated to intermediate or two or more... basis of the benefits accruing to intermediate and final cost objectives. When substantially the same...

  19. Scene independent real-time indirect illumination

    DEFF Research Database (Denmark)

    Frisvad, Jeppe Revall; Christensen, Niels Jørgen; Falster, Peter

    2005-01-01

    A novel method for real-time simulation of indirect illumination is presented in this paper. The method, which we call Direct Radiance Mapping (DRM), is based on basal radiance calculations and does not impose any restrictions on scene geometry or dynamics. This makes the method tractable for rea...

  20. Comparison of immunofluorescence and enzyme-linked immunosorbent assay and immunoglobulin G avidity techniques for screening of anti: Toxoplasma antibodies among single serum sample pregnant women in Tabriz, Iran

    Directory of Open Access Journals (Sweden)

    Mehrangiz Rajaii

    2015-01-01

    Full Text Available Background: Congenital toxoplasmosis is that pregnant women acquire the infection during gestation; diagnosis of the acute infection during pregnancy is a complex subject of maternal toxoplasmosis. Thus, the presence of immunoglobulin G (IgG and/or IgM Toxoplasma antibodies in a single serum sample drawn during gestation cannot be used to define whether the infection was recently acquired or chronic. Materials and Methods: At this cross-sectional descriptive study, sera of 391 pregnant women examined and compared. They were in an age range of 21-35 years, referred by gynecologists and infectious disease specialists, during March 2012-April 2013. They have referred, 215 (54.98%, 102 (26%, 74 (18.92% in the first, second and third trimesters of gestation, respectively. For each of them, a questionnaire was completed and serum samples were prepared in an equal condition, examined according to the procedures of indirect immunofluorescence (IIF, enzyme-linked immunosorbent assay (ELISA and IgG Avidity techniques. Results: We have found 111 (28.38% seronegative and 280 (71.61% seropositive cases by IIF and 124 (31.70% seronegative, 267 (68.28% seropositive cases by ELISA. The IgG avidity test confirmed 45 (69.23% and 7 (10.76% doubtful cases of IgM test in IIF and ELISA techniques. Conclusions: This study highlights how to manage pregnant women with toxoplasmosis, especially in a single serum sample condition.

  1. Molecular detection of Rickettsia typhi in cats and fleas.

    Directory of Open Access Journals (Sweden)

    Maria Mercedes Nogueras

    Full Text Available BACKGROUND: Rickettsiatyphi is the etiological agent of murine typhus (MT, a disease transmitted by two cycles: rat-flea-rat, and peridomestic cycle. Murine typhus is often misdiagnosed and underreported. A correct diagnosis is important because MT can cause severe illness and death. Our previous seroprevalence results pointed to presence of human R. typhi infection in our region; however, no clinical case has been reported. Although cats have been related to MT, no naturally infected cat has been described. The aim of the study is to confirm the existence of R. typhi in our location analyzing its presence in cats and fleas. METHODOLOGY/PRINCIPAL FINDINGS: 221 cats and 80 fleas were collected from Veterinary clinics, shelters, and the street (2001-2009. Variables surveyed were: date of collection, age, sex, municipality, living place, outdoor activities, demographic area, healthy status, contact with animals, and ectoparasite infestation. IgG against R. typhi were evaluated by indirect immunofluorescence assay. Molecular detection in cats and fleas was performed by real-time PCR. Cultures were performed in those cats with positive molecular detection. Statistical analysis was carried out using SPSS. A p < 0.05 was considered significant. Thirty-five (15.8% cats were seropositive. There were no significant associations among seropositivity and any variables. R. typhi was detected in 5 blood and 2 cultures. High titres and molecular detection were observed in stray cats and pets, as well as in spring and winter. All fleas were Ctenocephalides felis. R. typhi was detected in 44 fleas (55%, from shelters and pets. Co-infection with R. felis was observed. CONCLUSIONS: Although no clinical case has been described in this area, the presence of R. typhi in cats and fleas is demonstrated. Moreover, a considerable percentage of those animals lived in households. To our knowledge, this is the first time R. typhi is detected in naturally infected cats.

  2. Two distinct neural mechanisms underlying indirect reciprocity.

    Science.gov (United States)

    Watanabe, Takamitsu; Takezawa, Masanori; Nakawake, Yo; Kunimatsu, Akira; Yamasue, Hidenori; Nakamura, Mitsuhiro; Miyashita, Yasushi; Masuda, Naoki

    2014-03-18

    Cooperation is a hallmark of human society. Humans often cooperate with strangers even if they will not meet each other again. This so-called indirect reciprocity enables large-scale cooperation among nonkin and can occur based on a reputation mechanism or as a succession of pay-it-forward behavior. Here, we provide the functional and anatomical neural evidence for two distinct mechanisms governing the two types of indirect reciprocity. Cooperation occurring as reputation-based reciprocity specifically recruited the precuneus, a region associated with self-centered cognition. During such cooperative behavior, the precuneus was functionally connected with the caudate, a region linking rewards to behavior. Furthermore, the precuneus of a cooperative subject had a strong resting-state functional connectivity (rsFC) with the caudate and a large gray matter volume. In contrast, pay-it-forward reciprocity recruited the anterior insula (AI), a brain region associated with affective empathy. The AI was functionally connected with the caudate during cooperation occurring as pay-it-forward reciprocity, and its gray matter volume and rsFC with the caudate predicted the tendency of such cooperation. The revealed difference is consistent with the existing results of evolutionary game theory: although reputation-based indirect reciprocity robustly evolves as a self-interested behavior in theory, pay-it-forward indirect reciprocity does not on its own. The present study provides neural mechanisms underlying indirect reciprocity and suggests that pay-it-forward reciprocity may not occur as myopic profit maximization but elicit emotional rewards.

  3. Comparative efficacy of antigen and antibody detection tests for human trichinellosis

    International Nuclear Information System (INIS)

    Ivanoska, D.; Cuperlovic, K.; Gamble, H.R.; Murrell, K.D.

    1989-01-01

    Sera collected from patients with suspected or confirmed exposure to Trichinella spiralis were tested for circulating parasite antigens and antiparasite antibodies. Using an immunoradiometric assay, excretory--secretory antigens from muscle-stage larvae of T. spiralis were detected in the sera of 47% of 62 patients with clinical trichinellosis and 13% of 39 patients without clinical signs but suspected of exposure to infected meat. In comparison, antibodies were detected using an indirect immunofluorescent test in the circulation of 100% of the 62 patients with clinical trichinellosis and 46% of the 39 patients with suspected exposure. The presence of antibodies specific to excretory-secretory products of T. spiralis muscle larvae was confirmed in the majority of the samples tested by a monoclonal antibody-based competitive inhibition assay. These results indicate that antibody detection is a more sensitive diagnostic method for human trichinellosis, but that antigen detection might be a useful confirmatory test because it is a direct demonstration of parasite products in the circulation

  4. INDIRECT LABOR COSTS AND IMPLICATIONS FOR OVERHEAD ALLOCATION

    OpenAIRE

    Bea Chiang

    2013-01-01

    Cost accounting typically allocates indirect labor cost to cost object based on direct labor hours. The allocation process implicitly assumes that indirect labor costs vary proportionally with direct labor hours. The assumption of a linear relationship between indirect and direct labor is particularly suspicious at low production volume levels because there tends to be a fixed component in indirect labor. The linearity assumption is also challenged by recent increasing complexity of indirect ...

  5. An immunofluorescence assay for extracellular matrix components highlights the role of epithelial cells in producing a stable, fibrillar extracellular matrix

    Directory of Open Access Journals (Sweden)

    Omar S. Qureshi

    2017-10-01

    Full Text Available Activated fibroblasts are considered major drivers of fibrotic disease progression through the production of excessive extracellular matrix (ECM in response to signals from damaged epithelial and inflammatory cells. Nevertheless, epithelial cells are capable of expressing components of the ECM, cross-linking enzymes that increase its stability and are sensitive to factors involved in the early stages of fibrosis. We therefore wanted to test the hypothesis that epithelial cells can deposit ECM in response to stimulation in a comparable manner to fibroblasts. We performed immunofluorescence analysis of components of stable, mature extracellular matrix produced by primary human renal proximal tubular epithelial cells and renal fibroblasts in response to cytokine stimulation. Whilst fibroblasts produced a higher basal level of extracellular matrix components, epithelial cells were able to deposit significant levels of fibronectin, collagen I, III and IV in response to cytokine stimulation. In response to hypoxia, epithelial cells showed an increase in collagen IV deposition but not in response to the acute stress stimuli aristolochic acid or hydrogen peroxide. When epithelial cells were in co-culture with fibroblasts we observed significant increases in the level of matrix deposition which could be reduced by transforming growth factor beta (TGF-β blockade. Our results highlight the role of epithelial cells acting as efficient producers of stable extracellular matrix which could contribute to renal tubule thickening in fibrosis.

  6. Characterisation of L-Type Amino Acid Transporter 1 (LAT1 Expression in Human Skeletal Muscle by Immunofluorescent Microscopy

    Directory of Open Access Journals (Sweden)

    Nathan Hodson

    2017-12-01

    Full Text Available The branch chain amino acid leucine is a potent stimulator of protein synthesis in skeletal muscle. Leucine rapidly enters the cell via the L-Type Amino Acid Transporter 1 (LAT1; however, little is known regarding the localisation and distribution of this transporter in human skeletal muscle. Therefore, we applied immunofluorescence staining approaches to visualise LAT1 in wild type (WT and LAT1 muscle-specific knockout (mKO mice, in addition to basal human skeletal muscle samples. LAT1 positive staining was visually greater in WT muscles compared to mKO muscle. In human skeletal muscle, positive LAT1 staining was noted close to the sarcolemmal membrane (dystrophin positive staining, with a greater staining intensity for LAT1 observed in the sarcoplasmic regions of type II fibres (those not stained positively for myosin heavy-chain 1, Type II—25.07 ± 5.93, Type I—13.71 ± 1.98, p < 0.01, suggesting a greater abundance of this protein in these fibres. Finally, we observed association with LAT1 and endothelial nitric oxide synthase (eNOS, suggesting LAT1 association close to the microvasculature. This is the first study to visualise the distribution and localisation of LAT1 in human skeletal muscle. As such, this approach provides a validated experimental platform to study the role and regulation of LAT1 in human skeletal muscle in response to various physiological and pathophysiological models.

  7. Direct immunofluorescence assay compared to cell culture for the diagnosis of mucocutaneous herpes simplex virus infections in children.

    Science.gov (United States)

    Caviness, A Chantal; Oelze, Lindsay L; Saz, Ulas E; Greer, Jewel M; Demmler-Harrison, Gail J

    2010-09-01

    Direct immunofluorescence assay (DFA) is commonly used for the rapid identification of herpes simplex virus (HSV) infection in mucocutaneous lesions, yet little is known about its diagnostic accuracy. To determine the diagnostic yield and accuracy of HSV DFA for the diagnosis of mucocutaneous HSV infection in pediatric patients. Retrospective cross-sectional study of all patients who underwent HSV DFA testing by the Texas Children's Hospital Diagnostic Virology between January 1, 1995 and December 31, 2005. HSV DFA sensitivity, specificity, positive likelihood ratio (LRs), and negative LRs were estimated using viral culture as the reference standard. 659 specimens were submitted for HSV DFA with concurrent viral cultures. Viral cultures were positive for HSV type 1 in 158 (24%) and HSV type 2 in 2 (0.3%). There were 433 different patients with a median age of 8.6 years. Types of lesions were as follows: 50% ulcerative, 26% vesicular, 8% erythema or purpura, 5% pustular, and 11% missing. Of the 659 specimens submitted for HSV DFA, 160 (24%) were inconclusive due to inadequate cells. Of the 499 adequate specimens, overall HSV DFA test accuracy was: sensitivity 61%, specificity 99%, LR positive 40, and LR negative 0.39. A quarter of specimens submitted for HSV DFA testing are not adequate for DFA testing. When HSV DFA can be performed, it is specific, but not sensitive, for the identification of mucocutaneous HSV infection in children. Copyright (c) 2010 Elsevier B.V. All rights reserved.

  8. The Indirect cost of illness in Africa

    Directory of Open Access Journals (Sweden)

    Joses Muthuri Kirigia

    2018-06-01

    Full Text Available Background: The current study estimated (a the indirect costs associated with non-fatal disability and premature death across a wide range of diseases and health conditions in Africa in 2015 and (b the potential savings that could be accrued if countries were to meet the 3 health targets of the substainable development goal (SDG compared to the costs under the status quo. Methods: This study used the lost output or human capital approach to quantify the gross domestic product (GDP losses associated with the disability-adjusted-life-years (DALYs lost due to all causes by age group as well as by country economic classification (Group 1: 10 high/upper-middle income countries; Group 2: 17 lower-middle income countries; and Group 3: 27 low income countries. Results: The expected indirect cost of the 704,765,879 DALYs lost in Africa in 2015 was Int$ 2,983,187,560,197. Of this amount, 25.17%, 57.84% and 16.99% were incurred by the economies of the countries comprising Group 1, Group 2 and Group 3, respectively. Of the total continental indirect cost, 36.9%, 10.5%, 13.7%, 17.0%, 7.6%, 6.8% and 7.5% were associated with people aged 0-4, 5-14, 15-29, 30-49, 50-59, 60-69 and 70 years or older, respectively. Most of the total indirect cost (56.61% was attributable to maternal conditions, AIDS, tuberculosis (TB, malaria, neglected tropicald diseases (NTDs, non-communicable diseases and traffic injuries. Approximately half (47% of this cost could be avoided (or saved every year if the 3 (health targets of the SDG were fully met. Conclusion: The study estimated the total indirect cost of illness due to all causes by age group and country economic classification. The annual indirect cost is substantial. The findings contained in this paper suggest that health system strengthening should focus on both rich and poor countries, people of all ages and specific disease categories.

  9. How good is technetium-99m mercaptoacetyltriglycine indirect cystography?

    International Nuclear Information System (INIS)

    Sadeleer, C. de; Boe, V. de; Keuppens, F.; Desprechins, B.; Verboven, M.; Piepsz, A.

    1994-01-01

    The aim of the present study was to estimate the sensitivity of indirect radionuclide cystography (IRC) performed with technetium-99m mercaptoacetyltriglycine (MAG3), in comparison with the micturition cystourethrography (MCUG) and direct radionuclide cystography (DRC), for the diagnosis of vesicoureteric reflux. Two groups of patients were selected: group I comprised 40 children who underwent IRC during the acute phase of urinary tract infection and an MCUG 6 weeks later; group II comprised 42 (other) children with known reflux, who underwent IRC and DRC during follow-up. Taking as the reference the total number of refluxing kidneys detected by means of any cystographic technique, 99m Tc-MAG3 ICR missed two-thirds of the refluxing kidneys. Most of the small refluxes were missed, but so too were 50% of the major refluxes. Taking as the reference 99m Tc dimercaptosuccinic acid (DMSA) scintigraphy, MCUG detected 91% of the patients with DMSA abnormalities on at least one kidney, DRC detected 95%, and IRC detected 46% and 43% respectively, in groups I and II. The use of 99m Tc-MAG3 IRC as the sole technique for the detection of vesicoureteric reflux gives rise to an unacceptable number of false-negative results. (orig.)

  10. Cruel intentions on television and in real life: can viewing indirect aggression increase viewers' subsequent indirect aggression?

    Science.gov (United States)

    Coyne, Sarah M; Archer, John; Eslea, Mike

    2004-07-01

    Numerous studies have shown that viewing violence in the media can influence an individual's subsequent aggression, but none have examined the effect of viewing indirect aggression. This study examines the immediate effect of viewing indirect and direct aggression on subsequent indirect aggression among 199 children ages 11 to 14 years. They were shown an indirect, direct, or no-aggression video and their subsequent indirect aggression was measured by negative evaluation of a confederate and responses to a vignette. Participants viewing indirect or direct aggression gave a more negative evaluation of and less money to a confederate than participants viewing no-aggression. Participants viewing indirect aggression gave less money to the confederate than those viewing direct aggression. Participants viewing indirect aggression gave more indirectly aggressive responses to an ambiguous situation and participants viewing direct aggression gave more directly aggressive responses. This study provides the first evidence that viewing indirect aggression in the media can have an immediate impact on subsequent aggression.

  11. Indirect genetic effects and kin recognition

    DEFF Research Database (Denmark)

    Alemu, Setegn Worku; Berg, Peer; Janss, Luc

    2014-01-01

    Social interactions among individuals are widespread, both in natural and domestic populations. As a result, trait values of individuals may be affected by genes in other individuals, a phenomenon known as indirect genetic effects (IGEs). IGEs can be estimated using linear mixed models. The tradi......Social interactions among individuals are widespread, both in natural and domestic populations. As a result, trait values of individuals may be affected by genes in other individuals, a phenomenon known as indirect genetic effects (IGEs). IGEs can be estimated using linear mixed models...... present a reduced model that yields estimates of the total heritable effects on kin, on non-kin and on all social partners of an individual, as well as the total heritable variance for response to selection. Finally we discuss the consequences of analysing data in which IGEs depend on relatedness using...

  12. Lack of indirect sensitization in vivo

    International Nuclear Information System (INIS)

    Hetzel, F.W.; Avery, K.; Mensinger, M.; Frinak, S.; Tidwell, C.

    1985-01-01

    The possible utility of respiratory inhibiting drugs as indirect radiation sensitizers has been investigated in both in vitro and in vivo systems. In vitro studies were conducted in V79 monolayer and spheroid cultures examining both respiratory inhibition and radiation survival as end points. These drugs (BCNU, Mustargen and Chlorambucil) were found to be potent respiratory inhibitors and, in the spheroid system, to be effective indirect radiation sensitizers with enhancement ratios of approximately 2.0. In vivo studies with these drugs have also been conducted in a C/sub 3/H mouse MCa tumor system to determine if the same reoxygenation effect observed in spheroids could be demonstrated in the solid tumor system. Detailed microelectrode studies, employing each drug at its LD/sub 10/ level, have been unable to demonstrate any drug induced reoxygenation for any of the drugs tested. Complete details are presented

  13. Direct or indirect - that is the question!

    DEFF Research Database (Denmark)

    Pallesen, Ulla

    2016-01-01

    Time has passed since the prevention of caries was performed by extending cavity preparations wide in the tooth and below the gingiva. In the same period adhesive techniques have improved and nowadays made it possible to extend the indications for direct composite resin restorations, also when...... rather destroyed teeth need to be restored. Although much can be solved by direct fillings and thereby make treatments less invasive for the tooth and less expensive for the patient, there still will be teeth, where the prognosis is improved, if an indirect restoration in terms of an inlay, onlay...... or crown is made. Factors as remaining tooth structure, endodontic treatment, number of teeth, caries risk, para-functional habits, material properties, economy etc. will all have an influence, when the dentist guides the patient to choose the best solution. What is best for the tooth – direct or indirect...

  14. Social intervention and risk reduction - indirect countermeasures

    International Nuclear Information System (INIS)

    Higgins, N.A.; Morrey, M.

    1996-01-01

    An indirect countermeasure (IC) is an action which is intended to mitigate detrimental effects experienced by individuals or the community after an accident. Indirect countermeasures (ICs) achieve this, both by averting radiation risks arising from the accident, but by removing or reducing other risks and sources of stress or harm to which the community may be subject. ICs naturally fall into two categories: social action ICs, which range from introducing compensation payments to providing information centres; and risk reducing ICs which mitigate risks to which the population might be exposed, such as radon. By including a consideration of ICs in an assessment of the optimal response, it is likely that a decision maker will become aware of a greater range of harms and benefits that might result from the application of a countermeasure. The decision maker will then be in a better position to judge the appropriateness of any action. (author)

  15. Color indirect effects on melatonin regulation

    Science.gov (United States)

    Mian, Tian; Liu, Timon C.; Li, Yan

    2002-04-01

    Color indirect effect (CIE) is referred to as the physiological and psychological effects of color resulting from color vision. In previous papers, we have studied CIE from the viewpoints of the integrated western and Chinese traditional medicine, put forward the color-autonomic- nervous-subsystem model (CAM), and provided its time-theory foundation. In this paper, we applied it to study light effects on melatonin regulation in humans, and suggested that it is CIE that mediates light effects on melatonin suppression.

  16. Software Prefetching for Indirect Memory Accesses

    OpenAIRE

    Ainsworth, Sam; Jones, Timothy

    2017-01-01

    Many modern data processing and HPC workloads are heavily memory-latency bound. A tempting proposition to solve this is software prefetching, where special non-blocking loads are used to bring data into the cache hierarchy just before being required. However, these are difficult to insert to effectively improve performance, and techniques for automatic insertion are currently limited. This paper develops a novel compiler pass to automatically generate software prefetches for indirect mem...

  17. Draft of textbook focused on indirect taxes

    OpenAIRE

    Nováková, Hana

    2012-01-01

    The diploma thesis is about draft of textbook which is focused on indirect taxes for business school. The thesis is divided into theoretical and practical part. The theoretical part is focused on teaching tax issues at business school. Readers are introduced with basic studying documents, for example framework educational program and school educational program. In the following part I have conducted didactic analysis of curriculum and textbook of economics for business schools. The draft of t...

  18. Comment traduire en japonais les styles indirect et indirect libre de Madame Bovary ?

    Directory of Open Access Journals (Sweden)

    Hisaki Sawasaki

    2012-03-01

    Full Text Available Parmi les difficultés rencontrées lors de la traduction des textes littéraires occidentaux, en japonais, nous examinons le problème des styles indirect et indirect libre. Pour cela, en effectuant une petite mise au point grammaticale, nous comparons sept traductions de Madame Bovary de Gustave Flaubert, dont les dates de parution s’étendent sur une cinquantaine d’années. Cette période s’apparente, de notre point de vue, à un long itinéraire pour assimiler la notion occidentale des styles direct et indirect, tout en la conciliant avec les particularités du japonais. D’un autre côté, ce travail acharné des traducteurs a influencé quelque peu la langue japonaise. On trouve dans l’annexe tous les textes en japonais examinés.We will examine the difficulties met when translating Western literary texts in Japanese, in particular the problem of indirect and free indirect styles. We will define the grammatical issue and compare seven translations of Madame Bovary, published over a fifty year period. This time can be likened to a long path taken to digest the Western notion of direct and indirect styles, while reconciling it with Japanese language characteristics. On the other hand the translators’ relentless work has somewhat influenced the Japanese language. The annex will present all the Japanese texts examined.

  19. Indirect carotid cavernous fistula mimicking ocular myasthenia.

    Science.gov (United States)

    Leishangthem, Lakshmi; Satti, Sudhakar Reddy

    2017-10-19

    71-year-old woman with progressive left-sided, monocular diplopia and ptosis. Her symptoms mimicked ocular myasthenia, but she had an indirect carotid cavernous fistula (CCF). She was diagnosed with monocular myasthenia gravis (negative acetylcholinesterase antibody) after a positive ice test and started on Mestinon and underwent a thymectomy complicated by a brachial plexus injury. Months later, she developed left-sided proptosis and ocular bruit. She was urgently referred to neuro-interventional surgery and was diagnosed with an indirect high-flow left CCF, which was treated with Onyx liquid and platinum coil embolisation. Mestinon was discontinued. Her ophthalmic symptoms resolved. However, she was left with a residual left arm and hand hemiparesis and dysmetria secondary to a brachial plexus injury. Indirect CCF usually can present with subtle and progressive symptoms leading to delayed diagnosis or misdiagnosis. It is important for ophthalmologists to consider this differential in a patient with progressive ocular symptoms. © BMJ Publishing Group Ltd (unless otherwise stated in the text of the article) 2017. All rights reserved. No commercial use is permitted unless otherwise expressly granted.

  20. Indirect Comprehensive Review Board (ICRB). Final Report

    International Nuclear Information System (INIS)

    1996-12-01

    Lockheed Martin Idaho Technologies Company (LMITCO) used a systems engineering approach to take the first step toward defining a requirements baseline for all indirect work at the Idaho National Engineering Laboratory. The intent of this effort was to define the requirements for indirect work, identify the activities necessary to meet the requirements, and to produce defensible cost estimates for the work. The result of this effort is a scrubbed-down, defensible budget for all indirect work in FY 1997. Buying power for each dollar of direct work was increased by $.02. Recommendations are identified for improvements to this process in FY 1998. The purpose of this report is twofold. First is to report the final results of the 1996 ICRB process, and second is to document the process used such that incremental improvements may be made in future years. Objectives, processes, and approaches are described to provide a trail for future boards. Appendices contain copies of board composition, documentation of the process, as well as the actual training materials

  1. Interactive indirect illumination using adaptive multiresolution splatting.

    Science.gov (United States)

    Nichols, Greg; Wyman, Chris

    2010-01-01

    Global illumination provides a visual richness not achievable with the direct illumination models used by most interactive applications. To generate global effects, numerous approximations attempt to reduce global illumination costs to levels feasible in interactive contexts. One such approximation, reflective shadow maps, samples a shadow map to identify secondary light sources whose contributions are splatted into eye space. This splatting introduces significant overdraw that is usually reduced by artificially shrinking each splat's radius of influence. This paper introduces a new multiresolution approach for interactively splatting indirect illumination. Instead of reducing GPU fill rate by reducing splat size, we reduce fill rate by rendering splats into a multiresolution buffer. This takes advantage of the low-frequency nature of diffuse and glossy indirect lighting, allowing rendering of indirect contributions at low resolution where lighting changes slowly and at high-resolution near discontinuities. Because this multiresolution rendering occurs on a per-splat basis, we can significantly reduce fill rate without arbitrarily clipping splat contributions below a given threshold-those regions simply are rendered at a coarse resolution.

  2. Bose-Einstein condensation and indirect excitons: a review.

    Science.gov (United States)

    Combescot, Monique; Combescot, Roland; Dubin, François

    2017-06-01

    We review recent progress on Bose-Einstein condensation (BEC) of semiconductor excitons. The first part deals with theory, the second part with experiments. This Review is written at a time where the problem of exciton Bose-Einstein condensation has just been revived by the understanding that the exciton condensate must be dark because the exciton ground state is not coupled to light. Here, we theoretically discuss this missed understanding before providing its experimental support through experiments that scrutinize indirect excitons made of spatially separated electrons and holes. The theoretical part first discusses condensation of elementary bosons. In particular, the necessary inhibition of condensate fragmentation by exchange interaction is stressed, before extending the discussion to interacting bosons with spin degrees of freedom. The theoretical part then considers composite bosons made of two fermions like semiconductor excitons. The spin structure of the excitons is detailed, with emphasis on the crucial fact that ground-state excitons are dark: indeed, this imposes the exciton Bose-Einstein condensate to be not coupled to light in the dilute regime. Condensate fragmentations are then reconsidered. In particular, it is shown that while at low density, the exciton condensate is fully dark, it acquires a bright component, coherent with the dark one, beyond a density threshold: in this regime, the exciton condensate is 'gray'. The experimental part first discusses optical creation of indirect excitons in quantum wells, and the detection of their photoluminescence. Exciton thermalisation is also addressed, as well as available approaches to estimate the exciton density. We then switch to specific experiments where indirect excitons form a macroscopic fragmented ring. We show that such ring provides efficient electrostatic trapping in the region of the fragments where an essentially-dark exciton Bose-Einstein condensate is formed at sub-Kelvin bath

  3. Spatially indirect excitons in coupled quantum wells

    Energy Technology Data Exchange (ETDEWEB)

    Lai, Chih-Wei Eddy [Univ. of California, Berkeley, CA (United States)

    2004-03-01

    Microscopic quantum phenomena such as interference or phase coherence between different quantum states are rarely manifest in macroscopic systems due to a lack of significant correlation between different states. An exciton system is one candidate for observation of possible quantum collective effects. In the dilute limit, excitons in semiconductors behave as bosons and are expected to undergo Bose-Einstein condensation (BEC) at a temperature several orders of magnitude higher than for atomic BEC because of their light mass. Furthermore, well-developed modern semiconductor technologies offer flexible manipulations of an exciton system. Realization of BEC in solid-state systems can thus provide new opportunities for macroscopic quantum coherence research. In semiconductor coupled quantum wells (CQW) under across-well static electric field, excitons exist as separately confined electron-hole pairs. These spatially indirect excitons exhibit a radiative recombination time much longer than their thermal relaxation time a unique feature in direct band gap semiconductor based structures. Their mutual repulsive dipole interaction further stabilizes the exciton system at low temperature and screens in-plane disorder more effectively. All these features make indirect excitons in CQW a promising system to search for quantum collective effects. Properties of indirect excitons in CQW have been analyzed and investigated extensively. The experimental results based on time-integrated or time-resolved spatially-resolved photoluminescence (PL) spectroscopy and imaging are reported in two categories. (i) Generic indirect exciton systems: general properties of indirect excitons such as the dependence of exciton energy and lifetime on electric fields and densities were examined. (ii) Quasi-two-dimensional confined exciton systems: highly statistically degenerate exciton systems containing more than tens of thousands of excitons within areas as small as (10 micrometer)2 were

  4. Green Approach To Synthesize Crystalline Nanoscale ZnII-Coordination Polymers: Cell Growth Inhibition and Immunofluorescence Study.

    Science.gov (United States)

    Mukherjee, Somali; Ganguly, Sumi; Manna, Krishnendu; Mondal, Sanchaita; Mahapatra, Supratim; Das, Debasis

    2018-04-02

    Five new coordination polymers (CPs) namely, [{Zn(μ 2 -H 2 O) 0.5 (5N 3 -IPA)(2,2'-bpe)}] ∞ (1), [{Zn(μ 2 -H 2 O) 0.5 (5N 3 -IPA)(1,10-phen)}] ∞ (2), [{Zn(5N 3 -IPA)(1,2-bpe)}] ∞ (3), [{Zn(5N 3 -IPA)(1,2-bpey)}] ∞ (4), and [{Zn(H 2 O)(5N 3 -IPA)(4,4'-tme)}(H 2 O) 0.5 ] ∞ (5) (5N 3 -H 2 IPA = 5-azidoisophthalic acid, 2,2'-bpe= 2,2'-bipyridine, 1,10-phen = 1,10-phenanthroline, 1,2-bpe = 1,2-bis(4-pyridyl)ethane, 1,2-bpey = 1,2-bis(4-pyridyl)ethylene, 4,4'-tme = 4,4'-trimethylenedipyridine), have been synthesized based on a mixed ligand approach adopting a solvothermal technique. Depending upon the intrinsic structural flexibility of the bis-pyridyl coligands, interesting structural topologies have also been observed in the resulting CPs: Sra SrAl2 type topology for 3 and a 3-fold interpenetrated dmp topology for 4. A green hand grinding technique has been implemented to reduce the particle size of the CPs to generate nanoscale CPs (NCPs). SEM studies of NCPs reveal the formation of square and spherical particles for NCP 1 and 2, respectively, and nano rod for NCP 3, 4, and 5. Remarkably, when scaled down to nano range all the NCPs retain their crystalline nature. The cytotoxic activity of the NCPs (1-5) has been studied using human colorectal carcinoma cells (HCT 116). Significant cell death is observed for NCP 2, which is further corroborated by cell growth inhibition study. The observed cell death is likely to be due to mitochondrial-assisted apoptosis as is evident from immunofluorescence study.

  5. The extrinsic affective Simon task as an instrument for indirect assessment of prejudice

    NARCIS (Netherlands)

    Degner, J.; Wentura, D.

    2008-01-01

    We report one study that explored the applicability of the Extrinsic Affective Simon Task (EAST) as an indirect measure of prejudice. The EAST detected known differences in reactions revealing that a Turkish outgroup was spontaneously evaluated more negatively than the German ingroup. More

  6. Living in a noisy world: indirect effects of noise on animal communication

    NARCIS (Netherlands)

    Naguib, M.

    2013-01-01

    Noise is a ubiquitous feature in natural as well as in urban habitats. The presence of noise can have multiple direct and indirect effects on communication. Noise can directly mask signals leading to reduced detection and recognition. Noise also affects internal physiological processes which can

  7. Bioconjugated fluorescent silica nanoparticles for the rapid detection of Entamoeba histolytica.

    Science.gov (United States)

    Hemadi, Ahmad; Ekrami, Alireza; Oormazdi, Hormozd; Meamar, Ahmad Reza; Akhlaghi, Lame; Samarbaf-Zadeh, Ali Reza; Razmjou, Elham

    2015-05-01

    Rapid detection of Entamoeba histolytica based on fluorescent silica nanoparticle (FSNP) indirect immunofluorescence microscopy was evaluated. Silica nanoparticles were synthesized using Stöber's method, with their surface activated to covalently bind to, and immobilize, protein A. For biolabeling, FSNP was added to conjugated E. histolytica trophozoites with monoclonal anti-E. histolytica IgG1 for microscopic observation of fluorescence. Fluorescent silica nanoparticle sensitivity was determined with axenically cultured E. histolytica serially diluted to seven concentrations. Specificity was evaluated using other intestinal protozoa. Fluorescent silica nanoparticles detected E. histolytica at the lowest tested concentration with no cross-reaction with Entamoeba dispar, Entamoeba moshkovskii, Blastocystis sp., or Giardia lamblia. Visualization of E. histolytica trophozoites with anti-E. histolytica antibody labeled with fluorescein isothiocyanate (FITC) was compared with that using anti-E. histolytica antibody bioconjugated FSNP. Although FITC and FSNP produced similar results, the amount of specific antibody required for FITC to induce fluorescence of similar intensity was fivefold that for FSNP. Fluorescent silica nanoparticles delivered a rapid, simple, cost-effective, and highly sensitive and specific method of detecting E. histolytica. Further study is needed before introducing FSNP for laboratory diagnosis of amoebiasis. Copyright © 2015 Elsevier B.V. All rights reserved.

  8. Ma2 antibodies: an evaluation of commercially available detection methods.

    Science.gov (United States)

    Johannis, Wibke; Renno, Joerg H; Wielckens, Klaus; Voltz, Raymond

    2011-01-01

    Ma2 antibodies belong to the onconeuronal antibodies which define a "definite" paraneoplastic neurological syndrome (PNS). Because of the clinical relevance, use of two separate methods (indirect immunofluorescence technique--IFT--and immunoblot) is advocated; however, with an increasing number of commercially available assay systems, usually only one assay is performed. We compared IFT and three commercially available immunoblots (ravo Diagnostika, Euroimmun, Milenia Biotec) on sera from 35 patients with clinically suspected PNS. 17 were Ma2 antibody associated as defined by consensus result (showing positive reactivity in 2 assays), 18 were Ma2 antibody negative controls. Sensitivity/specificity for single assays were for IFT 94%/94%, for ravo Diagnostika PNS blot 88%/100%, for Euroimmun Neuronal Antigens Profile blot 100%/89%, and for Milenia Biotec MTR blot 94%/100%. Our data confirm, although all tests performed well, a combination of 2 independent assays is still advisable for Ma2 antibody detection in order to achieve higher sensitivity and specificity rates.

  9. The diagnostic value of indirect lower limb CT venography in the lower extremity deep venous thrombosis

    International Nuclear Information System (INIS)

    Zhu Li; Guo Youmin; Wang Jianguo; Guo Xiaojuan; Liu Min; Guo Yulin

    2009-01-01

    Objective: To evaluate indirect CT venography (CTV) in the diagnosis of deep venous thrombosis (DVT). Methods: A total of 516 acute pulmonary embolism patients proved by objective test were analyzed retrospectively. Using the results of lower limb compression sonography as reference standard, the sensitivity, specificity, positive predictive value and negative predictive value of indirect CTV were calculated. The agreement between lower extremity venous sonography and indirect CTV were assessed by Kappa analysis. Results: Among 516 patients with acute pulmonary thromboembolism (PTE), 110 patients underwent both CTV and CTPA and lower extremity sonography. DVT were detected by CTV and sonography simultaneously in 48 patients, while no DVT was detected in 39 patients by both examinations. Thirteen patients were diagnosed to have DVT by CTV without sonography detection, and DVT was detected in other 10 patients only by sonography. The sensitivity, specificity, positive predictive value and negative predictive value of indirect CTV was 82.8% (48/58), 75.0% (39/52), 78.7% (48/61) and 79.6% (39/49). The agreement between lower extremity venous sonography and indirect CTV was good for femoropopliteal vein (Kappa value range from 0.874 to 0.914, P=0.000). Thrombi detected by CTV were 62 and 52 respectively, and for sonography were 67 and 51 respectively. For calf veins, the agreement decreased (Kappa value range from 0.464 to 0.584, P=0.000). Thrombi detected by CTV were 6, 25 and 13 respectively, and for sonography were 13, 38 and 19 respectively. The agreement for external iliac vein was poor (Kappa value range from 0.230 to 0.262, P=0.067 and 0.004, respectively). Thrombi detected by CTV were 33, and for sonography were 17. Conclusions: The indirect CTV has high accuracy in the diagnosis of DVT. Combined CTPA and CTV can image pulmonary arteries and lower extremity veins in one examination. Indirect CTV can reveal thrombus in large pelvic veins, which has an advantage

  10. Indirect quantum tomography of quadratic Hamiltonians

    Energy Technology Data Exchange (ETDEWEB)

    Burgarth, Daniel [Institute for Mathematical Sciences, Imperial College London, London SW7 2PG (United Kingdom); Maruyama, Koji; Nori, Franco, E-mail: daniel@burgarth.de, E-mail: kmaruyama@riken.jp [Advanced Science Institute, RIKEN, Wako-shi, Saitama 351-0198 (Japan)

    2011-01-15

    A number of many-body problems can be formulated using Hamiltonians that are quadratic in the creation and annihilation operators. Here, we show how such quadratic Hamiltonians can be efficiently estimated indirectly, employing very few resources. We found that almost all the properties of the Hamiltonian are determined by its surface and that these properties can be measured even if the system can only be initialized to a mixed state. Therefore, our method can be applied to various physical models, with important examples including coupled nano-mechanical oscillators, hopping fermions in optical lattices and transverse Ising chains.

  11. Indirect evidence of calcitonin secretion in man.

    Science.gov (United States)

    Caniggia, A; Gennari, C; Vattimo, A; Nardi, P; Nuti, R

    1976-09-01

    1. The effect of calcitonin, a large amount of calcium given orally, pentagastrin and glucagon on plasma 47Ca radioactivity curves in subjects pretreated with 47Ca was examined. 2. A sudden increase of plasma radioactivity after intravenous administration of calcitonin and pentagastrin and after the oral calcium load was observed in normal subjects; the intravenous infusion of glucagon was less effective. 3. Two thyroparathyroidectomized patients who responded to the calcitonin infusion did not respond to the oral calcium load. 4. These data may be considered to offer indirect evidence of endogenous calcitonin secretion in man.

  12. Indirect Spectrophotometric Determination of Trace Quantities of Hydrazine

    Energy Technology Data Exchange (ETDEWEB)

    Shabani, A. M. Haji; Dadfarnia, S.; Dehghan, K. [Yazd University, Yazd (Iran, Islamic Republic of)

    2004-02-15

    An indirect, sensitive and accurate method for the determination of trace amounts of hydrazine is described. The method is based on the oxidation of hydrazine by a known excess of iodate in the presence of hydrochloric acid. The unreacted iodate is used in the oxidation of hydroxylamine to nitrite. Sulfanilic acid is diazotized by the nitrite formed. The resulting diazonium ion is coupled with N-(1-naphthyl)ethylenediamine to form a stable azo dye, which shows an absorption maximum at 540 nm. Hydrazine can be determined in the range of 20-400 ng mL{sup -1} with a detection limit of 3.1 ng mL{sup -1}. The relative standard deviation for 50, 200 and 400 ng mL{sup -1} of hydrazine is 2, 1.5 and 1.3%, respectively (n = 10). The method was applied to the determination of hydrazine in water samples

  13. Third European Company Survey – Direct and indirect employee participation

    NARCIS (Netherlands)

    Akkerman, Agnes; Sluiter, Roderick; Jansen, Giedo

    2015-01-01

    This report studies practices in EU establishments for direct and indirect employee participation in decision-making. Indirect employee participation is the involvement of employee representatives in decision-making processes, while direct employee participation describes direct interaction between

  14. Indirect Estimation of Selected Measures of Fertility and Marital ...

    African Journals Online (AJOL)

    DLHS6

    2018-01-09

    Jan 9, 2018 ... marital status distribution data of India especially of the 2011 census in deriving indirectly the fertility measures .... 2011 Census, Economic and Political weekly, EPW Vol. ... Indirect Estimates of Total Fertility Rate Using Child.

  15. THE INDIRECT OBJECT (IO) – ALBANIAN AND ENGLISH

    OpenAIRE

    Shkelqim Millaku; Xhevahire Topanica

    2016-01-01

    The aim of this study is too corporate the function of the indirect object between Albanian and English language. The function and the Albanian typical case for indirect object are dative and ablative. This grammatical phenomena is the full contrast between two languages because in English language doesn’t exist dative and ablative us in Albanian. In Albanian and English language, the indirect object is more heterogenic than the direct object. The indirect (direct) object in both of languages...

  16. On the Relative Size of Direct and Indirect Taxation

    OpenAIRE

    Soldatos, Gerasimos T.

    2014-01-01

    Modifying the standard analytical apparatuses for direct and indirect tax evasion to incorporate forward indirect tax shift in a monopolistically competitive environment, this paper maintains that indirect tax evasion would exceed for sure direct tax evasion only under consumer risk neutrality and a tax policy zeroing the tax shift. Also, in the presence of tax evasion, there cannot be optimal direct-indirect tax mix, because tax evasion is accompanied by uncertainty and hence, nonlinearities...

  17. [Detection of fps tumor antigen with mono-specific anti-fps serum in tumors induced by acute transforming ALV].

    Science.gov (United States)

    Wang, Yixin; Chen, Hao; Zhao, Peng; Li, Jianliang; Cui, Zhizhong

    2013-03-04

    To prepare anti-fps mono-specific serum, and detect the fps antigen in tumors induced by acute transforming avian leukosis/sarcoma virus containing v-fps oncogene. Two part of v-fps gene was amplified by RT-PCR using the Fu-J viral RNA as the template. Mono-specific serum was prepared by immuning Kunming white mouse with both two recombinant infusion proteins expressed by the prokaryotic expression system. Indirect immunofluorescent assay was used to detect fps antigen in tumor tissue suspension cells and CEF infected by sarcoma supernatant. Immunohistochemical method was used to detect fps antigen in tumor tissue. The mouse mono-specific serum was specific as it had no cross reaction with classical ALV-J strains. The result reveals that the tumor tissue suspension cells, the CEF infected by sarcoma supernatant, and the slice immunohistochemistry of the sarcoma showed positive results. The anti-fps mono-specific serum was prepared, and the detection method was established, which laid the foundation for the study of viral biological characteristics and mechanism of tumourgenesis of acute transforming avian leukosis/sarcoma virus containing v-fps oncogene.

  18. Discrepancies between feline coronavirus antibody and nucleic acid detection in effusions of cats with suspected feline infectious peritonitis.

    Science.gov (United States)

    Lorusso, Eleonora; Mari, Viviana; Losurdo, Michele; Lanave, Gianvito; Trotta, Adriana; Dowgier, Giulia; Colaianni, Maria Loredana; Zatelli, Andrea; Elia, Gabriella; Buonavoglia, Domenico; Decaro, Nicola

    2017-10-31

    Intra-vitam diagnosis of feline infectious peritonitis (FIP) is a challenge for veterinary diagnosticians, since there are no highly specific and sensitive assays currently available. With the aim to contribute to fill this diagnostic gap, a total of 61 effusions from cats with suspected effusive FIP were collected intra-vitam for detection of feline coronavirus (FCoV) antibodies and RNA by means of indirect immunofluorescence (IIF) assay and real-time RT-PCR (qRT-PCR), respectively. In 5 effusions there was no evidence for either FCoV RNA or antibodies, 51 and 52 specimens tested positive by IIF and qRT-PCR, respectively, although antibody titres≥1:1600, which are considered highly suggestive of FIP, were detected only in 37 effusions. Three samples with high antibody levels tested negative by qRT-PCR, whereas 18 qRT-PCR positive effusions contained no or low-titre antibodies. qRT-PCR positive samples with low antibody titres mostly contained low FCoV RNA loads, although the highest antibody titres were detected in effusions with C T values>30. In conclusion, combining the two methods, i.e., antibody and RNA detection would help improving the intra-vitam diagnosis of effusive FIP. Copyright © 2017 Elsevier Ltd. All rights reserved.

  19. Microdosimetric concepts for indirect radiation reactions

    International Nuclear Information System (INIS)

    Pohlit, W.

    1976-01-01

    It is the aim of microdosimetric models to get information about the influence of microscopic energy deposition by radiation on radiation reactions. Two parameters are always of interest: the energy necessary to produce a certain reaction and the volume in which this energy has to be desposited. In the simple case of an aqueous solution with point targets a complete description of radiation reactions in targets with various concentrations can be given. This model takes into consideration the distinct energy deposition on the particle track, diffusion of radicals and a mean energy necessary for a certain reaction at the target. Typical dose effect curves are obtained which allow the determination of all these quantities from experiments. The model has been tested with the oxydation of iron atoms in aqueous solutions and has been applied for more complex targets such as different alcohols and amino acids. The oxygen consumption in these reactions is used as an easy method for continuous measurement of these indirect reactions. It is the aim of such experiments to get enough quantitative microscopic data on indirect radiation reactions that these reactions can also be followed in living cells

  20. The indirect strategies of internationalization of companies

    Directory of Open Access Journals (Sweden)

    Đorđević Momčilo

    2012-01-01

    Full Text Available The subject of this work are, as mentioned in the title, the indirect strategies of internationalization of companies. The first part talks about the new business environment, the factors that lead to changes and their implications. It is at the same time the business context, which must be taken into consideration that determines the success of companies in the new business environment. In this paper, the emphasis is on company business in international markets, and the selection of appropriate strategies of internationalization, which will be responsible for this new business environment. There are many reasons that are discussed in this paper, and would rather indicate that partners from different countries to accept different strategies, rather than opting for a strategy of export, or a strategy of direct investments abroad. The dynamic technical and technological development, and its universal impact, among other relevant factors, put in the first place the importance of different strategies which is achieved through technology transfer, knowledge and experience whereby the exports, as a classic strategy of involvement in international trade, becomes significantly more difficult. Also, the internationalization of enterprises through direct investment abroad, involves many difficulties related to fitting in a foreign enterprise environment, and a high risk strategy. In addition, there is still a large number of countries do not allow wholly owned foreign enterprises. The paper discusses the value of various indirect internationalization strategy, which may be useful for those domestic companies that are in front of problem the choice of acceptable strategies of internationalization.

  1. Indirect costs of teaching in Canadian hospitals.

    Science.gov (United States)

    MacKenzie, T A; Willan, A R; Cox, M A; Green, A

    1991-01-01

    We sought to determine whether there are indirect costs of teaching in Canadian hospitals. To examine cost differences between teaching and nonteaching hospitals we estimated two cost functions: cost per case and cost per patient-day (dependent variables). The independent variables were number of beds, occupancy rate, teaching ratio (number of residents and interns per 100 beds), province, urbanicity (the population density of the county in which the hospital was situated) and wage index. Within each hospital we categorized a random sample of patient discharges according to case mix and severity of illness using age and standard diagnosis and procedure codes. Teaching ratio and case severity were each highly correlated positively with the dependent variables. The other variables that led to higher costs in teaching hospitals were wage rates and number of beds. Our regression model could serve as the basis of a reimbursement system, adjusted for severity and teaching status, particularly in provinces moving toward introducing case-weighting mechanisms into their payment model. Even if teaching hospitals were paid more than nonteaching hospitals because of the difference in the severity of illness there should be an additional allowance to cover the indirect costs of teaching. PMID:1898870

  2. Cooperation under indirect reciprocity and imitative trust.

    Science.gov (United States)

    Saavedra, Serguei; Smith, David; Reed-Tsochas, Felix

    2010-10-27

    Indirect reciprocity, a key concept in behavioral experiments and evolutionary game theory, provides a mechanism that allows reciprocal altruism to emerge in a population of self-regarding individuals even when repeated interactions between pairs of actors are unlikely. Recent empirical evidence show that humans typically follow complex assessment strategies involving both reciprocity and social imitation when making cooperative decisions. However, currently, we have no systematic understanding of how imitation, a mechanism that may also generate negative effects via a process of cumulative advantage, affects cooperation when repeated interactions are unlikely or information about a recipient's reputation is unavailable. Here we extend existing evolutionary models, which use an image score for reputation to track how individuals cooperate by contributing resources, by introducing a new imitative-trust score, which tracks whether actors have been the recipients of cooperation in the past. We show that imitative trust can co-exist with indirect reciprocity mechanisms up to a threshold and then cooperation reverses -revealing the elusive nature of cooperation. Moreover, we find that when information about a recipient's reputation is limited, trusting the action of third parties towards her (i.e. imitating) does favor a higher collective cooperation compared to random-trusting and share-alike mechanisms. We believe these results shed new light on the factors favoring social imitation as an adaptive mechanism in populations of cooperating social actors.

  3. Cooperation under indirect reciprocity and imitative trust.

    Directory of Open Access Journals (Sweden)

    Serguei Saavedra

    Full Text Available Indirect reciprocity, a key concept in behavioral experiments and evolutionary game theory, provides a mechanism that allows reciprocal altruism to emerge in a population of self-regarding individuals even when repeated interactions between pairs of actors are unlikely. Recent empirical evidence show that humans typically follow complex assessment strategies involving both reciprocity and social imitation when making cooperative decisions. However, currently, we have no systematic understanding of how imitation, a mechanism that may also generate negative effects via a process of cumulative advantage, affects cooperation when repeated interactions are unlikely or information about a recipient's reputation is unavailable. Here we extend existing evolutionary models, which use an image score for reputation to track how individuals cooperate by contributing resources, by introducing a new imitative-trust score, which tracks whether actors have been the recipients of cooperation in the past. We show that imitative trust can co-exist with indirect reciprocity mechanisms up to a threshold and then cooperation reverses -revealing the elusive nature of cooperation. Moreover, we find that when information about a recipient's reputation is limited, trusting the action of third parties towards her (i.e. imitating does favor a higher collective cooperation compared to random-trusting and share-alike mechanisms. We believe these results shed new light on the factors favoring social imitation as an adaptive mechanism in populations of cooperating social actors.

  4. 21 CFR 870.3640 - Indirect pacemaker generator function analyzer.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Indirect pacemaker generator function analyzer... Indirect pacemaker generator function analyzer. (a) Identification. An indirect pacemaker generator function analyzer is an electrically powered device that is used to determine pacemaker function or...

  5. System markets: Indirect network effects in action, or inaction?

    NARCIS (Netherlands)

    J.L.G. Binken (Jeroen)

    2010-01-01

    textabstractIn this dissertation, I empirically examine system markets up close. More specifically I examine indirect network effects, both demand-side and supply-side indirect network effects. Indirect network effects are the source of positive feedback in system markets, or so network effect

  6. Indirect Climatic Effects of Major Volcanic Eruptions

    Science.gov (United States)

    Hofmann, D. J.

    2007-05-01

    The direct effects on climate, related to atmospheric emissions to the atmosphere following major volcanic eruptions, are well-known although the sparseness of such eruptions make detailed study on the range of such variations difficult. In general terms, infrared absorption by volcanic emissions to the stratosphere result in local heating early in the event when gaseous sulfur compounds exist. This early period is followed by gas to particle conversion, on a time scale of 1-2 months, promoting the formation of sulfuric acid-water droplets. Coagulation and droplet growth result in the "volcanic stratospheric aerosol layer" which is related to the predominant direct climatic effect of large eruptions, the cooling of the troposphere by backscattering of solar visible radiation to space with a recovery time scale of 1-2 years. In this paper we will discuss some of the less-known "indirect" effects of the volcanic stratospheric aerosol on climate. We label them indirect as they act on climate through intermediary atmospheric constituents. The intermediaries in the volcanic indirect climatic effect are generally atmospheric greenhouse gases or other atmospheric gases and conditions which affect greenhouse gases. For example, cooling of the troposphere following major eruptions reduces the growth rate of atmospheric carbon dioxide related to respiration by the terrestrial biosphere. In addition, redirection of part of the direct solar beam into diffuse radiation by the volcanic stratospheric aerosol stimulates plant photosynthesis, further reducing the carbon dioxide growth rate. The growth rate of the second-most important atmospheric greenhouse gas, methane, is also affected by volcanic emissions. Volcanic stratospheric aerosol particles provide surface area which catalyzes heterogeneous chemical reactions thus stimulating removal of stratospheric ozone, also a greenhouse gas. Although major droughts usually related to ENSO events have opposite effects on carbon

  7. Detección de anticuerpos anti-Brucella spp. en cerdos mediante técnicas de aglutinación y ELISA indirecto en las provincias de Buenos Aires y La Pampa: Argentina Detection of anti-Brucella spp. antibodies in swine by agglutination techniques and indirect ELISA in the Buenos Aires and La Pampa provinces: Argentina

    Directory of Open Access Journals (Sweden)

    H.A. Castro

    2006-04-01

    Full Text Available En nuestro país no existe un programa de control sobre brucelosis porcina y su verdadera situación epidemiológica es desconocida. El objetivo de nuestro trabajo fue detectar la presencia de anticuerpos anti-Brucella spp. en porcinos provenientes de criaderos del sudoeste de la provincia de Buenos Aires y del este de la provincia de La Pampa. La toma de muestras de sangre se realizó en el momento del faenado de los animales. La detección de anticuerpos se efectuó mediante las técnicas de aglutinación con antígeno tamponado en placa (BPA, seroaglutinación en tubo (SAT, aglutinación con 2-ME (2-ME y ELISA indirecto, con dos antígenos diferentes: el antígeno CYT (fracción citoplasmática de B. abortus S19 y el antígeno CP (extracto citoplasmático libre de lipopolisacárido. Del total de las muestras analizadas (n=325, el 17,8% fue positivo para BPA, el 13,8% fue positivo para SAT y sólo el 8,0% fue positivo para 2-ME. Mediante ELISA-CYT, este porcentaje se elevó a 21,0%, mientras que a través del ELISA-CP sólo se halló un 10,0% de muestras reactivas. Estos resultados son compatibles con los informados en los escasos reportes previos para todo el país y sugieren la necesidad de extender los estudios a otras zonas, donde sea habitual la cría de cerdos.Porcine brucellosis is one of the most important zoonoses in this country. Currently, there is no control program for porcine brucellosis in Argentina and the epidemiological situation is still unknown. The purpose of our study was to detect anti-Brucella spp. antibodies in swine in the southwest of the Buenos Aires province and the east of the La Pampa province. Blood samples were obtained when animals were slaughtered. The presence of anti-brucella antibodies was studied by the buffered plate agglutination test (BPA, the tube agglutination test (SAT, the 2-mercaptoethanol (2-ME agglutination test and indirect ELISA tests, using the cytosolic fraction from Brucella abortus S19

  8. Comparison of the indirect fluorescent antibody test and modified agglutination test for detection of anti-Toxoplasma gondii antibodies in rats / Comparação da reação de imunofluorescência indireta e do teste de aglutinação modificado na detecção de anticorpos anti-Toxoplasma gondii em ratos

    Directory of Open Access Journals (Sweden)

    Roberta Lemos Freire

    2010-09-01

    Full Text Available The toxoplasmosis is a zoonosis caused by Toxoplasma gondii and affects a lot of species of carnivores and omnivores, including the human. The rodents are important in the transmition cycle because they act as an infection font to felines, the definitive host of this protozoan. The objective of this work was to evaluate the Modified Agglutination Test (MAT for the serologic diagnosis of toxoplasmosis in rats, comparing with the Indirect Fluorescent Antibody Test (IFAT, which has been considered the golden standard in animal toxoplasmosis diagnosis. Kappa test was used for comparing the serologic tests (IFAT and MAT and for determination of cutoff appropriate to MAT in this animal species. 182 rats were caught on local recycling of solid waste and solid residue storage in Londrina city, Paraná. Out of the 182 rats, nine (4.94% were positive to IFAT at a dilution of 1:16, and 17 (9.34% and five (2.75% were reactive to MAT in dilutions 1:25 and 1:50, respectively. The comparison of results between the techniques presented kappa coefficients of 0.26 and 0.55, respectively at 1:25 and 1:50 dilutions of MAT. It can be concluded that the dilution 1:50 is the most suitable to be used as cutoff for detecting T. gondii antibodies in rats using MAT, because agreed with IFAT.A toxoplasmose é uma zoonose causada pelo Toxoplasma gondii que acomete várias espécies carnívoras e onívoras, incluindo o ser humano. Os roedores são importantes na cadeia epidemiológica da doença por servirem de fonte de infecção aos felídeos, os hospedeiros definitivos deste protozoário. O objetivo deste trabalho foi avaliar o Teste de Aglutinação Modificada (MAT na detecção de anticorpos contra T. gondii em ratos, comparando-o à Reação de Imunofluorescência Indireta (RIFI, considerada padrão ouro para o diagnóstico da toxoplasmose animal. Empregou-se o teste kappa para a comparação dos testes sorológicos (RIFI e MAT e para a determinação do ponto de corte

  9. Indirect effects in community ecology: Their definition, study and importance.

    Science.gov (United States)

    Strauss, S Y

    1991-07-01

    The diversity of indirect interactions that can occur within communities is large. Recent research on indirect interactions is scattered in the literature under numerous labels. The definition of indirect effects is an important aspect of their study, and clarifies some of the subtle differences among indirect effects found in natural communities. Choosing which species to study, how to manipulate species and for what duration, which attributes to measure and, finally, which analytical techniques to use are all problems facing the community ecologist. Ultimately, we are striving for the best means of determining the relative importance of direct and indirect effects in structuring communities. Copyright © 1991. Published by Elsevier Ltd.

  10. Indirect evaporative coolers with enhanced heat transfer

    Science.gov (United States)

    Kozubal, Eric; Woods, Jason; Judkoff, Ron

    2015-09-22

    A separator plate assembly for use in an indirect evaporative cooler (IEC) with an air-to-air heat exchanger. The assembly includes a separator plate with a first surface defining a dry channel and a second surface defining a wet channel. The assembly includes heat transfer enhancements provided on the first surface for increasing heat transfer rates. The heat transfer enhancements may include slit fins with bodies extending outward from the first surface of separator plate or may take other forms including vortex generators, offset strip fins, and wavy fins. In slit fin implementations, the separator plate has holes proximate to each of the slit fins, and the separator plate assembly may include a sealing layer applied to the second surface of the separator plate to block air flow through the holes. The sealing layer can be a thickness of adhesive, and a layer of wicking material is applied to the adhesive.

  11. Haloperidol Selectively Remodels Striatal Indirect Pathway Circuits

    Science.gov (United States)

    Sebel, Luke E; Graves, Steven M; Chan, C Savio; Surmeier, D James

    2017-01-01

    Typical antipsychotic drugs are widely thought to alleviate the positive symptoms of schizophrenia by antagonizing dopamine D2 receptors expressed by striatal spiny projection neurons (SPNs). What is less clear is why antipsychotics have a therapeutic latency of weeks. Using a combination of physiological and anatomical approaches in ex vivo brain slices from transgenic mice, it was found that 2 weeks of haloperidol treatment induced both intrinsic and synaptic adaptations specifically within indirect pathway SPNs (iSPNs). Perphenazine treatment had similar effects. Some of these adaptations were homeostatic, including a drop in intrinsic excitability and pruning of excitatory corticostriatal glutamatergic synapses. However, haloperidol treatment also led to strengthening of a subset of excitatory corticostriatal synapses. This slow remodeling of corticostriatal iSPN circuitry is likely to play a role in mediating the delayed therapeutic action of neuroleptics. PMID:27577602

  12. Indirect Encoding in Neuroevolutionary Ship Handling

    Directory of Open Access Journals (Sweden)

    Miroslaw Lacki

    2018-03-01

    Full Text Available In this paper the author compares the efficiency of two encoding schemes for artificial intelligence methods used in the neuroevolutionary ship maneuvering system. This may be also be seen as the ship handling system that simulates a learning process of a group of artificial helmsmen - autonomous control units, created with an artificial neural network. The helmsman observes input signals derived form an enfironment and calculates the values of required parameters of the vessel maneuvering in confined waters. In neuroevolution such units are treated as individuals in population of artificial neural networks, which through environmental sensing and evolutionary algorithms learn to perform given task efficiently. The main task of this project is to evolve a population of helmsmen with indirect encoding and compare results of simulation with direct encoding method.

  13. Indirect adaptive control of discrete chaotic systems

    International Nuclear Information System (INIS)

    Salarieh, Hassan; Shahrokhi, Mohammad

    2007-01-01

    In this paper an indirect adaptive control algorithm is proposed to stabilize the fixed points of discrete chaotic systems. It is assumed that the functionality of the chaotic dynamics is known but the system parameters are unknown. This assumption is usually applicable to many chaotic systems, such as the Henon map, logistic and many other nonlinear maps. Using the recursive-least squares technique, the system parameters are identified and based on the feedback linearization method an adaptive controller is designed for stabilizing the fixed points, or unstable periodic orbits of the chaotic maps. The stability of the proposed scheme has been shown and the effectiveness of the control algorithm has been demonstrated through computer simulations

  14. Comparison of a direct and indirect ELISA for quantitating antisperm antibody in semen.

    Science.gov (United States)

    Lynch, D M; Howe, S E

    1987-01-01

    A direct and an indirect quantitative ELISA for antisperm antibody were compared using the spermatozoa and cell-free seminal fluid of 66 infertile males. The normal concentration of sperm binding immunoglobulin was less than or equal to 1.5 fg Ig per spermatozoon for the indirect seminal plasma assay and less than or equal to 1.5 fg Ig per spermatozoon by the direct assay. Of the 66 infertile males, 21% (14/66) had elevated levels of antisperm antibody in their seminal plasma and 26% (17/66) had elevated levels bound directly to their spermatozoa. The direct correlation between the results of these assays was 94%. A simple linear regression analysis between the indirect and direct measurements of antisperm antibody resulted in a correlation coefficient of r = 0.907. There was no statistically significant difference between results from the direct and indirect methods of the patients as a group. However, there was evidence of autospecificity in a small percentage of males who had elevated levels of antisperm antibody by the direct assay that was not detected by the indirect assay using pooled donor spermatozoa.

  15. Rapid antigen detection test for respiratory syncytial virus diagnosis as a diagnostic tool

    Directory of Open Access Journals (Sweden)

    Flávio da Silva Mesquita

    2017-05-01

    Full Text Available Objective: The aim of this study was to evaluate the QuickVue® RSV Test Kit (QUIDEL Corp, CA, USA as a screening tool for respiratory syncytial virus in children with acute respiratory disease in comparison with the indirect immunofluorescence assay as gold standard. In Brazil, rapid antigen detection tests for respiratory syncytial virus are not routinely utilized as a diagnostic tool, except for the diagnosis of dengue and influenza. Methods: The authors retrospectively analyzed 486 nasopharyngeal aspirate samples from children under age 5 with acute respiratory infection, between December 2013 and August 2014, the samples were analyzed by indirect immunofluorescence assay and QuickVue® RSV Test kit. Samples with discordant results were analyzed by real time PCR and nucleotide sequencing. Results: From 313 positive samples by immunofluorescence assays, 282 (90% were also positive by the rapid antigen detection test, two were positive only by rapid antigen detection test, 33 were positive only by immunofluorescence assays, and 171 were positive by both methods. The 35 samples with discordant results were analyzed by real time PCR; the two samples positive only by rapid antigen detection test and the five positive only by immunofluorescence assays were also positive by real time PCR. There was no relation between the negativity by QuickVue® RSV Test and viral load or specific strain. The QuickVue® RSV Test showed sensitivity of 90%, specificity of 98.8%, predictive positive value of 99.3%, and negative predictive value of 94.6%, with accuracy of 93.2% and agreement κ index of 0.85 in comparison to immunofluorescence assay. Conclusions: This study demonstrated that the QuickVue® RSV Test Kit can be effective in early detection of Respiratory syncytial virus in nasopharyngeal aspirate and is reliable for use as a diagnostic tool in pediatrics. Resumo: Objetivo: Avaliar o teste QuickVue® RSV Test Kit (QUIDEL Corp, CA, EUA para o diagn

  16. Fowl adenovirus serotype 4: Epidemiology, pathogenesis, diagnostic detection, and vaccine strategies.

    Science.gov (United States)

    Li, P H; Zheng, P P; Zhang, T F; Wen, G Y; Shao, H B; Luo, Q P

    2017-08-01

    Fowl adenovirus (FAdV) serotype-4 is highly pathogenic for chickens, especially for broilers aged 3 to 5 wk, and it has emerged as one of the foremost causes of economic losses to the poultry industry in the last 30 years. The liver is a major target organ of FAdV-4 infections, and virus-infected chickens usually show symptoms of hydropericardium syndrome. The virus is very contagious, and it is spread both vertically and horizontally. It can be isolated from infected liver homogenates and detected by several laboratory diagnostic methods (including an agar gel immunodiffusion test, indirect immunofluorescence assays, counterimmunoelectrophoresis, enzyme-linked immunosorbent assays, restriction endonuclease analyses, polymerase chain reaction (PCR), real-time PCR, and high-resolution melting-curve analyses). Although inactivated vaccines have been deployed widely to control the disease, attenuated live vaccines and subunit vaccines also have been developed, and they are more attractive vaccine candidates. This article provides a comprehensive review of FAdV-4, including its epidemiology, pathogenesis, diagnostic detection, and vaccine strategies. © 2017 Poultry Science Association Inc.

  17. Biological, serological and molecular techniques to xanthomonas axonopodis pv. Ci tri Asymptomatic detection

    International Nuclear Information System (INIS)

    Peyrou, M.; Del Campo, R.; Russi, P.; Mara, H.; Rigamonti, N.; Larrechart, L.; Perez, E.

    2012-01-01

    Xanthomonas axonopodis pv. ci tri (X ac) produces citrus canker disease in all citrus commercial species. The bacteria can be disseminated through vegetative propagation material in asymptomatic form. To optimize bacteria detection techniques applicable to asymptomatic citrus plant tissue routine analysis, ELISA, Immunofluorescence, Pcr, qRT P CR and host plant inoculation (bioassay) diagnostic techniques were compared. Tests were made from decimal dilutions between 108 uf6.ml-1 and 102 ufc.mL-1 using a pure culture of 49b strain.The detection level obtained was 1.8 x 102 ufc.mL-1 using Inmunofluorescence; 1.8x104 ufc.mL-1 with indirect ELISA, 1.8 x 103 ufc.mL-1 by means of PCR; 10 ufc.mL-1 through of qRT P CR and 230 ufc.mL-1 in sour orange inoculated plants. The experiment was repeated at least three times for each technique. Considering this result, and taking into account that sensitivity, practicity and cost, are important when a great number of plants need to be tested, the PCR and inoculation in host plants were those that met the best characteristics to be evaluated in asymptomatic plant material

  18. [Detection of serum anti-salivary duct antibody and its clinical significance].

    Science.gov (United States)

    Zhang, H; Shi, G Y; Cai, X H

    1990-11-01

    The authors developed an indirect immunofluorescence technique for the detection of Anti-salivary duct antibody (ASDA) and screened 34 patients with rheumatoid arthritis, 15 patients with Sjögren's syndrome-rheumatoid arthritis and 15 patients with primary Sjögren's syndrome, 63 cases with other connective tissue diseases, 9 cases with other diseases and 40 normal controls. The incidence of ASDA in patients with Sjögren's syndrome rheumatoid arthritis (66.67%) or rheumatoid arthritis (32.35%) was significantly higher than that in normal controls (P less than 0.001). In patients with primary Sjögren's syndrome and other CTDs, non-CTDs, no ASDA was found. However, in patients with Sjögren's syndrome-rheumatoid arthritis or rheumatoid arthritis alone, ASDA was not correlated with age, sex, disease duration or serological findings. The result suggests that the detection of serum ASDA might be useful in the differentiation of Sjögren's syndrome with rheumatoid arthritis from primary Sjögren's syndrome with arthralgia and/or arthritis.

  19. Novel genotype of Ehrlichia canis detected in samples of human blood bank donors in Costa Rica.

    Science.gov (United States)

    Bouza-Mora, Laura; Dolz, Gaby; Solórzano-Morales, Antony; Romero-Zuñiga, Juan José; Salazar-Sánchez, Lizbeth; Labruna, Marcelo B; Aguiar, Daniel M

    2017-01-01

    This study focuses on the detection and identification of DNA and antibodies to Ehrlichia spp. in samples of blood bank donors in Costa Rica using molecular and serological techniques. Presence of Ehrlichia canis was determined in 10 (3.6%) out of 280 blood samples using polymerase chain reaction (PCR) targeting the ehrlichial dsb conserved gene. Analysis of the ehrlichial trp36 polymorphic gene in these 10 samples revealed substantial polymorphism among the E. canis genotypes, including divergent tandem repeat sequences. Nucleotide sequences of dsb and trp36 amplicons revealed a novel genotype of E. canis in blood bank donors from Costa Rica. Indirect immunofluorescence assay (IFA) detected antibodies in 35 (35%) of 100 serum samples evaluated. Thirty samples showed low endpoint titers (64-256) to E. canis, whereas five sera yielded high endpoint titers (1024-8192); these five samples were also E. canis-PCR positive. These findings represent the first report of the presence of E. canis in humans in Central America. Copyright © 2016 Elsevier GmbH. All rights reserved.

  20. PM1-Alpha ELISA: the assay of choice for the detection of anti-PM/Scl autoantibodies?

    Science.gov (United States)

    Mahler, Michael; Fritzler, Marvin J

    2009-03-01

    A characteristic serological feature of patients suffering from the overlap polymyositis and scleroderma (PM/Scl) syndrome are antibodies to the human counterpart of the yeast exosome referred to as the PM/Scl complex. Historically, the detection of anti-PM/Scl antibodies was laborious and relied largely on indirect immunofluorescence and immunodiffusion techniques. In 1992 the major autoantigen PM/Scl-100 was identified and cloned. Subsequently, the major epitopes were mapped and one of these, termed PM1-Alpha, became the antigen for a novel ELISA exhibiting high sensitivity and specificity for the detection of anti-PM/Scl antibodies. Comparative studies with other methods using other PM/Scl autoantigens have shown that the PM1-Alpha ELISA has higher sensitivity and specificity than assays that employed recombinant PM/Scl-75c and PM/Scl-100. Anti-PM1-Alpha antibodies were identified in 55.0% of sera from PM/Scl overlap syndrome patients, but were also seen in 7.9% of SSc and in 7.5% of PM patients. The frequency in other systemic autoimmune diseases and in infectious diseases was significant lower. In summary, the data derived from individual studies suggest that PM1-Alpha may become the "gold standard" for the detection of anti-PM/Scl antibodies.

  1. Evaluation of the Chagas Stat-Paktm Assay for Detection of Trypanosoma cruzi Antibodies in Wildlife Reservoirs

    Science.gov (United States)

    Yabsley, Michael J.; Brown, Emily L.; Roellig, Dawn M.

    2010-01-01

    An immunochromatographic assay (Chagas Stat-Pak™) was evaluated for the detection of Trypanosoma cruzi antibodies in 4 species of wildlife reservoirs. Antibodies to T. cruzi were detected in raccoons (Procyon lotor) (naturally and experimentally infected) and degus (Octodon degu) (experimentally-infected) using the Chagas Stat-Pak. In naturally exposed wild raccoons, the Chagas Stat-Pak had a sensitivity and specificity of 66.7–80.0% and 96.3%, respectively. Compared with indirect immunofluorescent antibody assay results, serocon-version as determined by Chagas Stat-Pak was delayed for experimentally infected raccoons, but occurred sooner in experimentally infected degus. The Chagas Stat-Pak did not detect antibodies in naturally or experimentally infected Virginia opossums (Didelphis virginiana) or in experimentally infected short-tailed opossums (Monodelphis domestica). These data suggest that the Chagas Stat-Pak might be useful in field studies of raccoons and degus when samples would not be available for more-conventional serologic assays. Because this assay did not work on either species of marsupial, the applicability of the assay should be examined before it is used in other wild species. PMID:19016578

  2. Detection of anti-lactoferrin antibodies and anti-myeloperoxidase antibodies in autoimmune hepatitis: a retrospective study.

    Science.gov (United States)

    Tan, Liming; Zhang, Yuhong; Peng, Weihua; Chen, Juanjuan; Li, Hua; Ming, Feng

    2014-01-01

    Anti-lactoferrin antibodies (ALA) and anti-myeloperoxidase antibodies (AMPA) are specific serological markers for autoimmune hepatitis (AIH). The project aimed to detect ALA and AMPA and explore their clinical significances in AIH patients. 59 AIH patients, 217 non AIH patients, and 50 healthy controls were enrolled in this study. ALA and AMPA were detected by ELISA. Antineutropil cytoplasmic antibodies (ANCA) and anti-smooth muscle antibodies (ASMA) were examined by indirect immunofluorescence. Antimitochondrial antibody M2 subtype (AMA-M2), anti-liver kidney microsomal antibody Type 1 (LKM1), anti-liver cytosol antibody Type 1 (LC1), and anti-soluble liver antigen/liver-pancreas antibodies (SLA/LP) were tested by immunoblot. The positivity for ALA was 18.6% in AIH group, only one patient in non-AIH group was positive for ALA; the positivity for AMPA was 59.3% in AIH group, with significant differences (P < 0.01) compared with other groups. The specificities for ALA and AMPA were 99.63% and 97.75%; the sensitivities were 18.64% and 59.32%; and the accuracy rates were 84.97% and 90.80%, respectively. A certain correlation was observed between ALA and SLA/LP, AMPA and ANCA, ASMA in AIH group. ALA and AMPA were associated with AIH, and had high clinical diagnostic value. Co-detection with other relative autoantibodies could play an important role in differential diagnosis of AIH.

  3. Performance comparison of direct and indirect lighting systems

    International Nuclear Information System (INIS)

    Rubinstein, F.; Morse, O.; Clark, T.

    1993-01-01

    The performance of a retrofitted indirect lighting system was compared to the performance of a typical de-lamped direct lighting system in a partitioned office space. Power, illuminance and luminance measurements were made for the de-lamped direct lighting system and after installation of the indirect lighting system with various lamp and ballast combinations. Using the same lamps and ballasts, average workplace illuminance was slightly higher with indirect lighting than with direct lighting. With indirect lighting, workplace lumen efficacy was 4.5% lower due to the higher power draw of the lamps in the more open and cooler indirect fixtures. Indirect lighting with 36 watt T-8 lamps and electronic ballasts achieved an initial average workplace illuminance of 45 foot-candles in the partitioned office space at only 1.1 w/ft 2

  4. Direct and indirect punishment among strangers in the field.

    Science.gov (United States)

    Balafoutas, Loukas; Nikiforakis, Nikos; Rockenbach, Bettina

    2014-11-11

    Many interactions in modern human societies are among strangers. Explaining cooperation in such interactions is challenging. The two most prominent explanations critically depend on individuals' willingness to punish defectors: In models of direct punishment, individuals punish antisocial behavior at a personal cost, whereas in models of indirect reciprocity, they punish indirectly by withholding rewards. We investigate these competing explanations in a field experiment with real-life interactions among strangers. We find clear evidence of both direct and indirect punishment. Direct punishment is not rewarded by strangers and, in line with models of indirect reciprocity, is crowded out by indirect punishment opportunities. The existence of direct and indirect punishment in daily life indicates the importance of both means for understanding the evolution of cooperation.

  5. Indirect land use change and biofuel policy

    International Nuclear Information System (INIS)

    Kocoloski, Matthew; Griffin, W Michael; Matthews, H Scott

    2009-01-01

    Biofuel debates often focus heavily on carbon emissions, with parties arguing for (or against) biofuels solely on the basis of whether the greenhouse gas emissions of biofuels are less than (or greater than) those of gasoline. Recent studies argue that land use change leads to significant greenhouse gas emissions, making some biofuels more carbon intensive than gasoline. We argue that evaluating the suitability and utility of biofuels or any alternative energy source within the limited framework of plus and minus carbon emissions is too narrow an approach. Biofuels have numerous impacts, and policy makers should seek compromises rather than relying solely on carbon emissions to determine policy. Here, we estimate that cellulosic ethanol, despite having potentially higher life cycle CO 2 emissions (including from land use) than gasoline, would still be cost-effective at a CO 2 price of $80 per ton or less, well above estimated CO 2 mitigation costs for many alternatives. As an example of the broader approach to biofuel policy, we suggest the possibility of using the potential cost reductions of cellulosic ethanol relative to gasoline to balance out additional carbon emissions resulting from indirect land use change as an example of ways in which policies could be used to arrive at workable solutions.

  6. Indirectly cooled large thin superconducting CDF solenoid

    International Nuclear Information System (INIS)

    Kondo, Kunitaka; Mori, Shigeki; Yoshizaki, Ryozo; Saito, Ryusei; Asano, Katsuhiko.

    1985-01-01

    The manufacturing technique of the indirectly cooled large thin superconducting solenoid for the collider detector facility (CDF solenoid) has been studied through cooperation of University of Tsukuba and the National Laboratory for High Energy Physics of the Ministry of Education of Japan, and the Fermi National Accelerator Laboratory in the U.S. Fabrication and testing of the solenoid has recently been completed by Hitachi. The CDF solenoid has a large-sized thin structure for meeting the requirement by experiments to be applied. Hitachi has thus developed a variety of new technologies including the design standard, coil cooling method, material selection, and manufacturing technique in accordance with experimental data, which were confirmed in a series of analyses and tests made on various prototypes. The CDF solenoid, built using Hitachi's new technologies, is of the world's top class among equipment of this type. This paper outlines the design criteria for the major components employed in the CDF solenoid and the test results of the solenoid. (author)

  7. INDIRECT WRITTEN CORRECTIVE FEEDBACK, REVISION, AND LEARNING

    Directory of Open Access Journals (Sweden)

    Fatemeh Poorebrahim

    2017-01-01

    Full Text Available Corrective feedback, the necessity of providing it, and how it should be provided has been one of the hot topics in the area of ELT. Amid continuing controversies over whether providing feedback helps L2 learners improve their writing accuracy, many research studies have been undertaken to compare the relative effectiveness of different types of feedback. However, the difference between two types of indirect corrective feedback, namely indication and indication plus location, have not been properly examined yet. Motivated to narrow this gap, this study is designed to compare two groups of Iranian learners, each revising their papers based on one of the aforementioned options. For data analysis, a series of independent samples t tests were employed. The results revealed that the difference between the two groups in their reduction of errors from the original draft to the revision of each task followed a growing trend and became significant. Nonetheless, the difference in accuracy of new pieces of writing fell short of significance. Finally, it was found that error reduction in revision stage cannot be considered as learning. The results of the study, discussed in relation to that of others, implicate that the purpose for which feedback is provided is essential in determining the type of feedback; more explicit feedback is better for revising purposes while more implicit feedback is good for learning purposes.

  8. Indirect Sensitivities to the Scale of Supersymmetry

    CERN Document Server

    Ellis, Jonathan Richard; Olive, Keith A; Weiglein, Georg

    2005-01-01

    Precision measurements, now and at a future linear electron-positron collider (ILC), can provide indirect information about the possible scale of supersymmetry. We illustrate the present-day and possible future ILC sensitivities within the constrained minimal supersymmetric extension of the Standard Model (CMSSM), in which there are three independent soft supersymmetry-breaking parameters m_{1/2}, m_0 and A_0. We analyze the present and future sensitivities separately for M_W, sin^2(theta_eff), (g-2)_mu, BR(b -> s gamma), BR(B_s -> mu+ mu-), M_h and Higgs branching ratios. We display the observables as functions of m_{1/2}, fixing m_0 so as to obtain the cold dark matter density allowed by WMAP and other cosmological data for specific values of A_0, tan beta and mu > 0. In a second step, we investigate the combined sensitivity of the currently available precision observables, M_W, sin^2(theta_eff), (g-2)_mu and BR(b -> s gamma), by performing a chi^2 analysis. The current data are in very good agreement with ...

  9. Theoretical research on color indirect effects

    Science.gov (United States)

    Liu, T. C.; Liao, Changjun; Liu, Songhao

    1995-05-01

    Color indirect effects (CIE) means the physiological and psychological effects of color resulting from color vision. In this paper, we study CIE from the viewpoints of the integrated western and Chinese traditional medicine and the time quantum theory established by C. Y. Liu et al., respectively, and then put forward the color-automatic-nervous-subsystem model that could color excites parasympathetic subsystem and hot color excites sympathetic subsystem. Our theory is in agreement with modern color vision theory, and moreover, it leads to the resolution of the conflict between the color code theory and the time code theory oncolor vision. For the latitude phenomena on athlete stars number and the average lifespan, we also discuss the possibility of UV vision. The applications of our theory lead to our succeeding in explaining a number of physiological and psychological effects of color, in explaining the effects of age on color vision, and in explaining the Chinese chromophototherapy. We also discuss its application to neuroimmunology. This research provides the foundation of the clinical applications of chromophototherapy.

  10. Decentralized indirect methods for learning automata games.

    Science.gov (United States)

    Tilak, Omkar; Martin, Ryan; Mukhopadhyay, Snehasis

    2011-10-01

    We discuss the application of indirect learning methods in zero-sum and identical payoff learning automata games. We propose a novel decentralized version of the well-known pursuit learning algorithm. Such a decentralized algorithm has significant computational advantages over its centralized counterpart. The theoretical study of such a decentralized algorithm requires the analysis to be carried out in a nonstationary environment. We use a novel bootstrapping argument to prove the convergence of the algorithm. To our knowledge, this is the first time that such analysis has been carried out for zero-sum and identical payoff games. Extensive simulation studies are reported, which demonstrate the proposed algorithm's fast and accurate convergence in a variety of game scenarios. We also introduce the framework of partial communication in the context of identical payoff games of learning automata. In such games, the automata may not communicate with each other or may communicate selectively. This comprehensive framework has the capability to model both centralized and decentralized games discussed in this paper.

  11. Indirect land use change and biofuel policy

    Science.gov (United States)

    Kocoloski, Matthew; Griffin, W. Michael; Matthews, H. Scott

    2009-09-01

    Biofuel debates often focus heavily on carbon emissions, with parties arguing for (or against) biofuels solely on the basis of whether the greenhouse gas emissions of biofuels are less than (or greater than) those of gasoline. Recent studies argue that land use change leads to significant greenhouse gas emissions, making some biofuels more carbon intensive than gasoline. We argue that evaluating the suitability and utility of biofuels or any alternative energy source within the limited framework of plus and minus carbon emissions is too narrow an approach. Biofuels have numerous impacts, and policy makers should seek compromises rather than relying solely on carbon emissions to determine policy. Here, we estimate that cellulosic ethanol, despite having potentially higher life cycle CO2 emissions (including from land use) than gasoline, would still be cost-effective at a CO2 price of 80 per ton or less, well above estimated CO2 mitigation costs for many alternatives. As an example of the broader approach to biofuel policy, we suggest the possibility of using the potential cost reductions of cellulosic ethanol relative to gasoline to balance out additional carbon emissions resulting from indirect land use change as an example of ways in which policies could be used to arrive at workable solutions.

  12. Noninvasive wearable sensor for indirect glucometry.

    Science.gov (United States)

    Zilberstein, Gleb; Zilberstein, Roman; Maor, Uriel; Righetti, Pier Giorgio

    2018-04-02

    A noninvasive mini-sensor for blood glucose concentration assessment has been developed. The monitoring is performed by gently pressing a wrist or fingertip onto the chemochromic mixture coating a thin glass or polymer film positioned on the back panel of a smart watch with PPG/HRM (photoplethysmographic/heart rate monitoring sensor). The various chemochromic components measure the absolute values of the following metabolites present in the sweat: acetone, acetone beta-hydroxybutirate, aceto acetate, water, carbon dioxide, lactate anion, pyruvic acid, Na and K salts. Taken together, all these parameters give information about blood glucose concentration, calculated via multivariate analysis based on neural network algorithms built into the sensor. The Clarke Error Grid shows an excellent correlation between data measured by the standard invasive glucose analyser and the present noninvasive sensor, with all points aligned along a 45-degree diagonal and contained almost exclusively in sector A. Graphs measuring glucose levels five times a day (prior, during and after breakfast and prior, during and after lunch), for different individuals (males and females) show a good correlation between the two curves of conventional, invasive meters vs. the noninvasive sensor, with an error of ±15%. This novel, noninvasive sensor for indirect glucometry is fully miniaturized, easy to use and operate and could represent a valid alternative in clinical settings and for individual, personal users, to current, invasive tools. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. Bioelectrochemical Systems for Indirect Biohydrogen Production

    KAUST Repository

    Regan, John M.

    2014-01-01

    Bioelectrochemical systems involve the use of exoelectrogenic (i.e., anode-reducing) microbes to produce current in conjunction with the oxidation of reduced compounds. This current can be used directly for power in a microbial fuel cell, but there are alternate uses of this current. One such alternative is the production of hydrogen in a microbial electrolysis cell (MEC), which accomplishes cathodic proton reduction with a slight applied potential by exploiting the low redox potential produced by exoelectrogens at the anode. As an indirect approach to biohydrogen production, these systems are not subject to the hydrogen yield constraints of fermentative processes and have been proven to work with virtually any biodegradable organic substrate. With continued advancements in reactor design to reduce the system internal resistance, increase the specific surface area for anode biofilm development, and decrease the material costs, MECs may emerge as a viable alternative technology for biohydrogen production. Moreover, these systems can also incorporate other value-added functionalities for applications in waste treatment, desalination, and bioremediation.

  14. System markets: Indirect network effects in action, or inaction?

    OpenAIRE

    Binken, Jeroen

    2010-01-01

    textabstractIn this dissertation, I empirically examine system markets up close. More specifically I examine indirect network effects, both demand-side and supply-side indirect network effects. Indirect network effects are the source of positive feedback in system markets, or so network effect theory tells us. Systems are composed of complementary and interdependent products, such as hardware and software. For instance, a video game system is composed of the video game console, on the one han...

  15. Indirect Catastrophic Injuries in Olympic Styles of Wrestling in Iran

    OpenAIRE

    Kordi, Ramin; Ziaee, Vahid; Rostami, Mohsen; Wallace, W. Angus

    2011-01-01

    Background: Data on indirect catastrophic injuries in wrestling are scarce. Objectives: To develop a profile of indirect catastrophic injuries in international styles of wrestling and to describe possible risk factors. Study Design: Retrospective case series; Level of evidence, 3. Methods: Indirect catastrophic injuries that occurred in wrestling clubs in Iran from July 1998 to June 2005 were identified by contacting several sources. The cases were retrospectively reviewed. Results: The injur...

  16. Tipping and Concentration in Markets with Indirect Network Effects

    OpenAIRE

    Jean-Pierre H. Dubé; Günter J. Hitsch; Pradeep K. Chintagunta

    2010-01-01

    This paper develops a framework for measuring “tipping”—the increase in a firm's market share dominance caused by indirect network effects. Our measure compares the expected concentration in a market to the hypothetical expected concentration that would arise in the absence of indirect network effects. In practice, this measure requires a model that can predict the counterfactual market concentration under different parameter values capturing the strength of indirect network effects. We build...

  17. Cell-based quantification of biomarkers from an ultra-fast microfluidic immunofluorescent staining: application to human breast cancer cell lines

    Science.gov (United States)

    Migliozzi, D.; Nguyen, H. T.; Gijs, M. A. M.

    2018-02-01

    Immunohistochemistry (IHC) is one of the main techniques currently used in the clinics for biomarker characterization. It consists in colorimetric labeling with specific antibodies followed by microscopy analysis. The results are then used for diagnosis and therapeutic targeting. Well-known drawbacks of such protocols are their limited accuracy and precision, which prevent the clinicians from having quantitative and robust IHC results. With our work, we combined rapid microfluidic immunofluorescent staining with efficient image-based cell segmentation and signal quantification to increase the robustness of both experimental and analytical protocols. The experimental protocol is very simple and based on fast-fluidic-exchange in a microfluidic chamber created on top of the formalin-fixed-paraffin-embedded (FFPE) slide by clamping it a silicon chip with a polydimethyl siloxane (PDMS) sealing ring. The image-processing protocol is based on enhancement and subsequent thresholding of the local contrast of the obtained fluorescence image. As a case study, given that the human epidermal growth factor receptor 2 (HER2) protein is often used as a biomarker for breast cancer, we applied our method to HER2+ and HER2- cell lines. We report very fast (5 minutes) immunofluorescence staining of both HER2 and cytokeratin (a marker used to define the tumor region) on FFPE slides. The image-processing program can segment cells correctly and give a cell-based quantitative immunofluorescent signal. With this method, we found a reproducible well-defined separation for the HER2-to-cytokeratin ratio for positive and negative control samples.

  18. Immunofluorescence Microscopy and mRNA Analysis of Human Embryonic Stem Cells (hESCs) Including Primary Cilia Associated Signaling Pathways

    DEFF Research Database (Denmark)

    Vestergaard, Maj Linea; Awan, Aashir; Warzecha, Caroline Becker

    2016-01-01

    onto 16-well glass chambers, and continuing with the general IFM and qPCR anlysis. The techniques are illustrated with results on cellular localization of transcriptional factors and components of the Hedgehog, Wnt, PDGF, and TGFβ signaling pathways to primary cilia in stem cell maintenance......This chapter describes the procedures for immunofluorescence microscopy (IFM) and quantitative PCR (qPCR) analyses of human embryonic stem cells (hESCs) grown specifically under feeder-free conditions. A detailed protocol is provided outlining the steps from initially growing the cells, passaging...

  19. Dew drops on spider web appearance: a newly named pattern of IgG4 deposition in pemphigus with direct immunofluorescence

    Directory of Open Access Journals (Sweden)

    Marian Dmochowski

    2017-08-01

    Full Text Available Novel appearances in cutaneous pathology as well as mucocutaneous clinical signs are being described which indicate that this is still an attractive area for exploration. The H + E histology terms of “decorated tomb stoning” and “undecorated tomb stoning”, advocated by some pathologists, are misleading and as such should be avoided. Here, an appearance of IgG4 pemphigus deposits examined cost-effectively with direct immunofluorescence and suggested to be called “dew drops on spider web” is depicted in depth.

  20. Comparison of the marginal adaptation of direct and indirect composite inlay restorations with optical coherence tomography.

    Science.gov (United States)

    Türk, Ayşe Gözde; Sabuncu, Metin; Ünal, Sena; Önal, Banu; Ulusoy, Mübin

    2016-01-01

    The purpose of the study was to use the photonic imaging modality of optical coherence tomography (OCT) to compare the marginal adaptation of composite inlays fabricated by direct and indirect techniques. Class II cavities were prepared on 34 extracted human molar teeth. The cavities were randomly divided into two groups according to the inlay fabrication technique. The first group was directly restored on cavities with a composite (Esthet X HD, Dentsply, Germany) after isolating. The second group was indirectly restored with the same composite material. Marginal adaptations were scanned before cementation with an invisible infrared light beam of OCT (Thorlabs), allowing measurement in 200 µm intervals. Restorations were cemented with a self-adhesive cement resin (SmartCem2, Dentsply), and then marginal adaptations were again measured with OCT. Mean values were statistically compared by using independent-samples t-test and paired samples t-test (pmarginal discrepancy values than indirect inlays, before (p=0.00001442) and after (p=0.00001466) cementation. Marginal discrepancy values were increased for all restorations after cementation (p=0.00008839, p=0.000000952 for direct and indirect inlays, respectively). The mean marginal discrepancy value of the direct group increased from 56.88±20.04 µm to 91.88±31.7 µm, whereas the indirect group increased from 107.54±35.63 µm to 170.29±54.83 µm. Different techniques are available to detect marginal adaptation of restorations, but the OCT system can give quantitative information about resin cement thickness and its interaction between tooth and restoration in a nondestructive manner. Direct inlays presented smaller marginal discrepancy than indirect inlays. The marginal discrepancy values were increased for all restorations that refer to cement thickness after cementation.

  1. An Information-Theoretic Approach for Indirect Train Traffic Monitoring Using Building Vibration

    OpenAIRE

    Xu, Susu; Zhang, Lin; Zhang, Pei; Noh, Hae Young

    2017-01-01

    This paper introduces an indirect train traffic monitoring method to detect and infer real-time train events based on the vibration response of a nearby building. Monitoring and characterizing traffic events are important for cities to improve the efficiency of transportation systems (e.g., train passing, heavy trucks, and traffic). Most prior work falls into two categories: (1) methods that require intensive labor to manually record events or (2) systems that require deployment of dedicated ...

  2. Detection of adrenocortical autoantibodies in Addison's disease with a peroxidase-labelled protein A technique

    Directory of Open Access Journals (Sweden)

    R.C. Silva

    1998-09-01

    Full Text Available Adrenocortical autoantibodies (ACA, present in 60-80% of patients with idiopathic Addison's disease, are conventionally detected by indirect immunofluorescence (IIF on frozen sections of adrenal glands. The large-scale use of IIF is limited in part by the need for a fluorescence microscope and the fact that histological sections cannot be stored for long periods of time. To circumvent these restrictions we developed a novel peroxidase-labelled protein A (PLPA technique for the detection of ACA in patients with Addison's disease and compared the results with those obtained with the classical IIF assay. We studied serum samples from 90 healthy control subjects and 22 patients with Addison's disease, who had been clinically classified into two groups: idiopathic (N = 13 and granulomatous (N = 9. ACA-PLPA were detected in 10/22 (45% patients: 9/13 (69% with the idiopathic form and 1/9 (11% with the granulomatous form, whereas ACA-IIF were detected in 11/22 patients (50%: 10/13 (77% with the idiopathic form and 1/9 (11% with the granulomatous form. Twelve of the 13 idiopathic addisonians (92% were positive for either ACA-PLPA or ACA-IIF, but only 7 were positive by both methods. In contrast, none of 90 healthy subjects was found to be positive for ACA. Thus, our study shows that the PLPA-based technique is useful, has technical advantages over the IIF method (by not requiring the use of a fluorescence microscope and by permitting section storage for long periods of time. However, since it is only 60% concordant with the ACA-IIF method, it should be considered complementary instead of an alternative method to IIF for the detection of ACA in human sera.

  3. Detection of viruses and atypical bacteria associated with acute respiratory infection of children in Hubei, China.

    Science.gov (United States)

    Wu, Zegang; Li, Yan; Gu, Jian; Zheng, Hongyun; Tong, Yongqing; Wu, Qing

    2014-02-01

    Acute respiratory infection is the major cause of disease and death in children, particularly in developing countries. However, the spectrum of pathogenic viruses and atypical bacteria that exist in many of these countries remains incompletely characterized. The aim of this study was to examine the spectrum of pathogenic viruses and atypical bacteria associated with acute respiratory infection in children under the age of 16. A total of 10 435 serum sera specimens were collected from hospitalized children presenting with acute respiratory infection symptoms. Indirect immunofluorescence assays were performed to detect immunoglobulin M antibodies against nine common pathogens: mycoplasma pneumonia, influenza virus B, respiratory syncytial virus, parainfluenza virus, adenovirus, influenza virus A, legionella pneumophila, coxiella burnetii and chamydophila pneumonia. Of the 10 435 specimens examined, 7046 tested positive for at least one pathogen. Among all of the tested pathogens, mycoplasma pneumonia had the highest detection rate (56.9%). Influenza virus A and influenza virus B epidemics occurred during both winter and summer. The detection rate of respiratory syncytial virus and adenovirus was higher in spring. Cases of mixed infection were more complex: 4136 specimens (39.6%) tested positive for ≥2 pathogens. There were statistically significant difference in detection rates of mycoplasma pneumonia, influenza virus B, respiratory syncytial virus, parainfluenza virus, adenovirus, influenza virus A, legionella pneumophila and chamydophila pneumonia among different age groups (P acute respiratory infection among children in Hubei of China were mycoplasma pneumonia, influenza virus B and respiratory syncytial virus. The detection rates for each pathogen displayed specific seasonal and age group variations. © 2013 The Authors. Respirology © 2013 Asian Pacific Society of Respirology.

  4. Detection of liver kidney microsomal type 1 antibody using molecularly based immunoassays.

    Science.gov (United States)

    Kerkar, N; Ma, Y; Davies, E T; Cheeseman, P; Mieli-Vergani, G; Vergani, D

    2002-12-01

    To assess the diagnostic value of two commercial molecularly based immunoassays detecting liver kidney microsomal type 1 antibody (LKM1). The performance of Varelisa and LKM1 enzyme linked immunosorbent assay (ELISA) was compared with immunofluorescence, and two validated research techniques-an in house ELISA and a radioligand assay measuring antibodies to P4502D6. Thirty serum samples from three patients with autoimmune hepatitis type 2 covering immunofluorescence titres of 1/10 to 1/10 240 and 55 LKM1 negative controls were tested. All 30 sera that were LKM1 positive by immunofluorescence were positive by the in house ELISA, the radioligand assay, and LKM1-ELISA, and 29 were also positive by Varelisa. None of the 55 sera negative for LKM1 by immunofluorescence was positive by the in house ELISA and radioligand assay, but one was positive by Varelisa and 14 were positive using the LKM1-ELISA. Agreement between immunofluorescence, the in house ELISA, the radioligand assay, and Varelisa was high (kappa > 0.8), and agreement between immunofluorescence and LKM1-ELISA was moderate (kappa = 0.63). The assay kit marketed as Varelisa allows accurate detection of LKM1.

  5. 42 CFR 412.322 - Indirect medical education adjustment factor.

    Science.gov (United States)

    2010-10-01

    ... Payment System for Inpatient Hospital Capital Costs Basic Methodology for Determining the Federal Rate for Capital-Related Costs § 412.322 Indirect medical education adjustment factor. (a) Basic data. CMS.... The indirect teaching adjustment factor equals [e (raised to the power of .2822×the ratio of residents...

  6. A Computational Approach to the Interpretation of Indirect Speech Acts

    NARCIS (Netherlands)

    Beun, R.J.; Eijk, R.M. van; Meyer, J-J.Ch.; Vergunst, N.L.

    2006-01-01

    An Indirect Speech Act (ISA) is an utterance that conveys a message that is different from its literal meaning, often for reasons of politeness or subtlety. The DenK-system provides us with a non-compositional way to look at Indirect Speech Acts that contain modal verbs. We can extract the

  7. Indirect Positive Evidence in the Acquisition of a Subset Grammar

    Science.gov (United States)

    Schwartz, Misha; Goad, Heather

    2017-01-01

    This article proposes that second language learners can use indirect positive evidence (IPE) to acquire a phonological grammar that is a subset of their L1 grammar. IPE is evidence from errors in the learner's L1 made by native speakers of the learner's L2. It has been assumed that subset grammars may be acquired using direct or indirect negative…

  8. The Adoption of Indirect Instruments of Monetary Policy

    OpenAIRE

    Tomás J. T. Baliño; Charles Enoch; William E. Alexander

    1995-01-01

    This paper examines the experience of implementing indirect instruments of monetary policy. The experiences of country studies illustrate the variety of circumstances under which indirect instruments of monetary policy have been introduced. Case Studies are presented for Chile, Egypt, Ghana, Indonesia, Mexico, New Zealand, and Poland.

  9. The Influence of Direct and Indirect Speech on Source Memory

    Directory of Open Access Journals (Sweden)

    Anita Eerland

    2018-02-01

    Full Text Available People perceive the same situation described in direct speech (e.g., John said, “I like the food at this restaurant” as more vivid and perceptually engaging than described in indirect speech (e.g., John said that he likes the food at the restaurant. So, if direct speech enhances the perception of vividness relative to indirect speech, what are the effects of using indirect speech? In four experiments, we examined whether the use of direct and indirect speech influences the comprehender’s memory for the identity of the speaker. Participants read a direct or an indirect speech version of a story and then addressed statements to one of the four protagonists of the story in a memory task. We found better source memory at the level of protagonist gender after indirect than direct speech (Exp. 1–3. When the story was rewritten to make the protagonists more distinctive, we also found an effect of speech type on source memory at the level of the individual, with better memory after indirect than direct speech (Exp. 3–4. Memory for the content of the story, however, was not influenced by speech type (Exp. 4. While previous research showed that direct speech may enhance memory for how something was said, we conclude that indirect speech enhances memory for who said what.

  10. 10 CFR 602.15 - Indirect cost limitations.

    Science.gov (United States)

    2010-01-01

    ... 10 Energy 4 2010-01-01 2010-01-01 false Indirect cost limitations. 602.15 Section 602.15 Energy DEPARTMENT OF ENERGY (CONTINUED) ASSISTANCE REGULATIONS EPIDEMIOLOGY AND OTHER HEALTH STUDIES FINANCIAL ASSISTANCE PROGRAM § 602.15 Indirect cost limitations. Awards issued under this part for conferences and...

  11. 10 CFR 605.16 - Indirect cost limitations.

    Science.gov (United States)

    2010-01-01

    ... 10 Energy 4 2010-01-01 2010-01-01 false Indirect cost limitations. 605.16 Section 605.16 Energy DEPARTMENT OF ENERGY (CONTINUED) ASSISTANCE REGULATIONS THE OFFICE OF ENERGY RESEARCH FINANCIAL ASSISTANCE PROGRAM § 605.16 Indirect cost limitations. Awards issued under this part for conferences and scientific...

  12. Indirect Control for Demand Side Management – A Conceptual Introduction

    DEFF Research Database (Denmark)

    Heussen, Kai; You, Shi; Biegel, Benjamin

    2012-01-01

    The concept of “indirect control” has become a relevant discussion term in relation to activation distributed and small-scale demand and generation units to provide resources for power system balancing. The term and its association with price signals has, however caused some confusion as to its...... correct definition, either as a control or a market concept. This paper aims to provide a conceptual introduction to "indirect control" for management of small and distributed demand side resources. A review of control concepts and an analysis of "indirectness" features are provided to create a framework...... for systematic classification of indirect control strategies. The concepts developed then enable a discussion of control performance and valuation of direct- and indirect control strategies....

  13. Smoking Cessation Is Associated With Lower Indirect Costs.

    Science.gov (United States)

    Baker, Christine L; Bruno, Marianna; Emir, Birol; Li, Vicky W; Goren, Amir

    2018-06-01

    This study quantified differences in indirect costs due to decreased work productivity between current and former smokers. Former smokers were further categorized by number of years since quitting to assess corresponding differences. Data on employed individuals were obtained from the 2013 US National Health and Wellness Survey (NHWS; N = 75,000). Indirect costs were calculated for current smokers and former smokers from weekly wages based on age and sex. The annual total indirect costs for current smokers were $1327.53, $1560.18, and $1839.87 higher than for those who quit 0 to 4 years, 5 to 10 years, and more than or equal to 11 years prior, respectively. There were no significant differences in mean total indirect costs between the former smoker groups. Current smokers showed significantly higher total annual indirect costs compared with former smokers, independently of the number of years since quitting smoking.

  14. Forecasting of indirect consumables for a Job Shop

    Science.gov (United States)

    Shakeel, M.; Khan, S.; Khan, W. A.

    2016-08-01

    A job shop has an arrangement where similar machines (Direct consumables) are grouped together and use indirect consumables to produce a product. The indirect consumables include hack saw blades, emery paper, painting brush etc. The job shop is serving various orders at a particular time for the optimal operation of job shop. Forecasting is required to predict the demand of direct and indirect consumables in a job shop. Forecasting is also needed to manage lead time, optimize inventory cost and stock outs. The objective of this research is to obtain the forecast for indirect consumables. The paper shows how job shop can manage their indirect consumables more accurately by establishing a new technique of forecasting. This results in profitable use of job shop by multiple users.

  15. An indecent proposal: the dual functions of indirect speech.

    Science.gov (United States)

    Chakroff, Aleksandr; Thomas, Kyle A; Haque, Omar S; Young, Liane

    2015-01-01

    People often use indirect speech, for example, when trying to bribe a police officer by asking whether there might be "a way to take care of things without all the paperwork." Recent game theoretic accounts suggest that a speaker uses indirect speech to reduce public accountability for socially risky behaviors. The present studies examine a secondary function of indirect speech use: increasing the perceived moral permissibility of an action. Participants report that indirect speech is associated with reduced accountability for unethical behavior, as well as increased moral permissibility and increased likelihood of unethical behavior. Importantly, moral permissibility was a stronger mediator of the effect of indirect speech on likelihood of action, for judgments of one's own versus others' unethical action. In sum, the motorist who bribes the police officer with winks and nudges may not only avoid public punishment but also maintain the sense that his actions are morally permissible. Copyright © 2014 Cognitive Science Society, Inc.

  16. Development of indirect indicators for CO2 intrusion into freshwater

    International Nuclear Information System (INIS)

    Humez, P.

    2012-01-01

    This study deals with the impact of CO 2 leakages out of geological storage into overlying freshwater aquifers. Compared to other existing studies, the major added value of this study lies, on the one hand, in the research of new monitoring tools and isotopic approach in the context of CCS aiming at early and sensitive detection of CO 2 leakage and, on the other hand, in the application of these tools at the (limited) laboratory scale as well as at field scale. In order to test these tools, solid and liquid materials were sampled out of the major strategic drinking water Albian aquifer in the Paris Basin (France). We have then precisely characterized and used them within a batch experiment. This experiment yields interesting results which help understanding and constraining precisely the water-rock-CO 2 interactions as well as the isotopic responses. A real scale application of the method was then performed in Norway. It was an opportunity to develop this isotopic program and to track the isotopic evolution composition, while differentiating the natural processes and the system response to the CO 2 injection. When applied to the detection of CO 2 leakage context, the two case studies open the way for choosing the 'best' isotopic tools as indirect indicators of CO 2 presence in these specific systems. The efficiency of these isotopic tools comes from the recording of the CO 2 footprint all along the water-rock-CO 2 interactions. Using such tools imposes a rigorous methodology, which is tackled in this manuscript. Furthermore, future application will require adapting to the specifics of a proposed site. (author)

  17. A novel indirect ELISA for diagnosis of dengue fever

    Directory of Open Access Journals (Sweden)

    Rohan Narayan

    2016-01-01

    Full Text Available Background & objectives: Dengue fever (DF is associated with significant morbidity and mortality in the tropical and sub-tropical regions of the world. Since there are no effective antiviral drugs for treatment, clinicians often rely on the accurate diagnosis of dengue fever to begin supportive therapy at early stages of the illness. The objective of this study was to develop an in-house dengue virus serotype 2 (DENV-2 non-structural protein- 5 (NS5 based indirect ELISA. Methods: DENV-2 was raised in Vero cells and the viral proteins were separated and subsequently the NS5 protein was eluted. Serum samples from primary and secondary dengue fever patients; and acute and convalescent samples from Japanese encephalitis (JE and West Nile virus (WNV cases were used to validate the ELISA. Results: The assay was found to be 100 per cent specific in detecting DENV-2 specific antibodies from patient′s serum. However, in terms of sensitivity, the assay could detect IgM antibodies only from 90 per cent of the primary dengue samples. The IgM/IgG ratio of the primary and secondary samples was 7.24 and 0.64, respectively. Interpretation & conclusions: The results indicate that the DENV-2 NS5 ELISA is dengue group specific and can be used to differentiate dengue infection from other circulating Flavivirus infections. This NS5 ELISA can also be used to distinguish between primary and secondary dengue fever on the basis of IgM/IgG ratios. Further studies with larger sample sizes and different DENV serotypes are required to validate the ELISA.

  18. Differing results of direct and indirect solid phase radioimmunoassay for HBsAg in acute hepatitis

    International Nuclear Information System (INIS)

    Strohm, W.D.; Legler, K.; Gerlich, W.; Stamm, B.; Zimmer, S.; Biotest-Serum-Institut G.m.b.H., Frankfurt am Main; Goettingen Univ.

    1978-01-01

    In 54 patients suffering from active viral hepatitis the indirect solid phase radioimmunoassay (ind-SPRIA) for HBsAg was positive in 9 cases the direct solid phase radioimmunoassay (d-SPRIA) being negative. In 2 further cases ind-SPRIA was positive during several weeks but d-SPRIA only once. AntiHBc could be detected in 9 of these patients. In 7 patients the usual decrease of the transaminase activity was followed by a second elavation with prolongation of the disease. The unknown factor detected by ind-SPRIA suggests a special of acute hepatitis. (orig.) [de

  19. Differing results of direct and indirect solid phase radioimmunoassay for HBsAg in acute hepatitis

    Energy Technology Data Exchange (ETDEWEB)

    Strohm, W D; Legler, K; Gerlich, W; Stamm, B; Zimmer, S [Frankfurt Univ. (Germany, F.R.). Abt. fuer Gastroenterologie; Biotest-Serum-Institut G.m.b.H., Frankfurt am Main (Germany, F.R.); Goettingen Univ. (Germany, F.R.). Hygiene-Institut)

    1978-09-01

    In 54 patients suffering from active viral hepatitis the indirect solid phase radioimmunoassay (ind-SPRIA) for HBsAg was positive in 9 cases the direct solid phase radioimmunoassay (d-SPRIA) being negative. In 2 further cases ind-SPRIA was positive during several weeks but d-SPRIA only once. AntiHBc could be detected in 9 of these patients. In 7 patients the usual decrease of the transaminase activity was followed by a second elavation with prolongation of the disease. The unknown factor detected by ind-SPRIA suggests a special of acute hepatitis.

  20. Detection of serum antibodies against Bartonella species in cats with sporotrichosis from Rio de Janeiro, Brazil.

    Science.gov (United States)

    Kitada, Amanda A B; Favacho, Alexsandra R M; Oliveira, Raquel V C; Pessoa, Adonai A; Gomes, Raphael; Honse, Carla O; Gremião, Isabella D F; Lemos, Elba R S; Pereira, Sandro A

    2014-04-01

    Cat scratch disease is a zoonosis caused by Bartonella species, transmitted to humans through scratches or bites from infected cats and via direct contact with infected feces. Sporotrichosis, caused by the fungal complex Sporothrix, is transmitted by traumatic inoculation of the fungus. Cats are important in zoonotic transmission. Serum samples from 112 domestic cats with sporotrichosis and 77 samples from healthy cats were analyzed by indirect immunofluorescence assay (IFA), using the commercial kit Bartonella henselae IFA IgG (Bion). The presence of antibodies against feline leukemia virus (FeLV) and of feline immunodeficiency virus (FIV) core antigens was detected using the commercial kit Snap Combo FIV-FeLV (Idexx). The group of animals with sporotrichosis contained 93 males with a median age of 22 months, eight (7.1%) of which were positive for FIV and 15 (13.4%) for FeLV. The group of animals without sporotrichosis contained 36 males with a median age 48 months, 10 (13.0%) of which were positive for FIV and eight (10.4%) for FeLV. Of the 112 cats with sporotrichosis and 77 cats without mycosis, 72 (64.3%) and 35 (45.5%), respectively, were IFA reactive. No association was found between age, sex, FIV/FeLV and the presence of antibodies to Bartonella species. The results suggest that the study population can be considered a potential source of zoonotic infection for both diseases.

  1. Standardization of serological tests for detecting anti-Trypanosoma cruzi antibodies in dogs

    Directory of Open Access Journals (Sweden)

    M. A. Lauricella

    1993-09-01

    Full Text Available This paper reports on the standardization of four serological reactions currently used in human serodiagnosis for the detection of anti-Trypanosoma cruzi antibodies in naturally and experimentally infected dogs. Indirect immunofluorescence test (IFAT and hemagglutination test (IHAT were standardized, and complement fixation test (CFT and direct agglutination test (DAT were used for diagnostic confirmation. Four hundred and eighty one mongrel dogs that were studied by xenodiagnosis were used: (1 parasitemic dogs of two localities of endemic area (EA of Santiago del Estero province in Argentina (n = 134; (2 non-parasitemic dogs of the same area (n = 285; (3 dogs experimentally infected with T. cruzi in the patent period (n = 6; (4 non-infected dogs (n = 56 which were born in the city of Buenos Aires (BA, one non-EA for Chagas' disease. For IFAT, parasitemic dogs EA showed 95% of reactive sera. Non parasitemic dogs EA showed 77% of non reactive sera. None sera from BA were reactive for dilutions higher than four. For IHAT, 84% of sera of parasitemic dogs EA showed serological reactivity and among non parasitemic dogs BA, 61% were non reactive, while the remainder showed at most titres of 1/16. The cut-off titres for IFAT and IHAT were 1/16 and 1/32 respectively, and for CFT and DAT 1/1 and 1/128 respectively. Sensitivity for IFAT, IHAT, CF and DAT were 95%, 84%, 97% and 95% respectively.

  2. Sensitivity and specificity of parallel or serial serological testing for detection of canine Leishmania infection

    Directory of Open Access Journals (Sweden)

    Mauro Maciel de Arruda

    2016-01-01

    Full Text Available In Brazil, human and canine visceral leishmaniasis (CVL caused byLeishmania infantum has undergone urbanisation since 1980, constituting a public health problem, and serological tests are tools of choice for identifying infected dogs. Until recently, the Brazilian zoonoses control program recommended enzyme-linked immunosorbent assays (ELISA and indirect immunofluorescence assays (IFA as the screening and confirmatory methods, respectively, for the detection of canine infection. The purpose of this study was to estimate the accuracy of ELISA and IFA in parallel or serial combinations. The reference standard comprised the results of direct visualisation of parasites in histological sections, immunohistochemical test, or isolation of the parasite in culture. Samples from 98 cases and 1,327 noncases were included. Individually, both tests presented sensitivity of 91.8% and 90.8%, and specificity of 83.4 and 53.4%, for the ELISA and IFA, respectively. When tests were used in parallel combination, sensitivity attained 99.2%, while specificity dropped to 44.8%. When used in serial combination (ELISA followed by IFA, decreased sensitivity (83.3% and increased specificity (92.5% were observed. Serial testing approach improved specificity with moderate loss in sensitivity. This strategy could partially fulfill the needs of public health and dog owners for a more accurate diagnosis of CVL.

  3. Antigen detection systems

    Science.gov (United States)

    Infectious agents or their constituent parts (antigens or nucleic acids) can be detected in fresh, frozen, or fixed tissues or other specimens, using a variety of direct or indirect assays. The assays can be modified to yield the greatest sensitivity and specificity but in most cases a particular m...

  4. An improved recommendation algorithm via weakening indirect linkage effect

    International Nuclear Information System (INIS)

    Chen Guang; Qiu Tian; Shen Xiao-Quan

    2015-01-01

    We propose an indirect-link-weakened mass diffusion method (IMD), by considering the indirect linkage and the source object heterogeneity effect in the mass diffusion (MD) recommendation method. Experimental results on the MovieLens, Netflix, and RYM datasets show that, the IMD method greatly improves both the recommendation accuracy and diversity, compared with a heterogeneity-weakened MD method (HMD), which only considers the source object heterogeneity. Moreover, the recommendation accuracy of the cold objects is also better elevated in the IMD than the HMD method. It suggests that eliminating the redundancy induced by the indirect linkages could have a prominent effect on the recommendation efficiency in the MD method. (paper)

  5. Types of suggestibility: Relationships among compliance, indirect, and direct suggestibility.

    Science.gov (United States)

    Polczyk, Romuald; Pasek, Tomasz

    2006-10-01

    It is commonly believed that direct suggestibility, referring to overt influence, and indirect suggestibility, in which the intention to influence is hidden, correlate poorly. This study demonstrates that they are substantially related, provided that they tap similar areas of influence. Test results from 103 students, 55 women and 48 men, were entered into regression analyses. Indirect suggestibility, as measured by the Sensory Suggestibility Scale for Groups, and compliance, measured by the Gudjonsson Compliance Scale, were predictors of direct suggestibility, assessed with the Barber Suggestibility Scale. Spectral analyses showed that indirect suggestibility is more related to difficult tasks on the BSS, but compliance is more related to easy tasks on this scale.

  6. Indirect Taxes in Romania – an Econometric Analysis

    Directory of Open Access Journals (Sweden)

    Daniela Penu

    2016-03-01

    Full Text Available Indirect taxes have a significant weight in the GDP. Statistical data reveals that the new states that joined the EU in its funding sources to the greatest extent through indirect taxes. They reveal, on the one hand, reducing the tax burden on the producer, and on the other hand, the difficulty faced by the country in direct tax collection, but also some problems of social nature. This article proposes an econometric analysis of the relationship between indirect taxes and household final consumption expenditure.

  7. Indirect Genetic Effects for group-housed animals

    DEFF Research Database (Denmark)

    Alemu, Setegn Worku

    This thesis investigated social interactions in group-housed animals. The main findings of this thesis: 1) Statistical methods to estimate indirect genetic effects when interactions differ between kin vs. non-kin were developed. 2) Indirect genetic effects contribute a substantial amount...... of heritable variation for bite mark traits in group-housed min. 3) Indirect genetic effects estimation needs to take into account systematic interactions due to sex or kin for bite mark trait in group-housed min. 4) Genomic selection can be used to increase the response to selection for survival time in Brown...

  8. Mediation analysis to estimate direct and indirect milk losses due to clinical mastitis in dairy cattle.

    Science.gov (United States)

    Detilleux, J; Kastelic, J P; Barkema, H W

    2015-03-01

    Milk losses associated with mastitis can be attributed to either effects of pathogens per se (i.e., direct losses) or effects of the immune response triggered by intramammary infection (indirect losses). The distinction is important in terms of mastitis prevention and treatment. Regardless, the number of pathogens is often unknown (particularly in field studies), making it difficult to estimate direct losses, whereas indirect losses can be approximated by measuring the association between increased somatic cell count (SCC) and milk production. An alternative is to perform a mediation analysis in which changes in milk yield are allocated into their direct and indirect components. We applied this method on data for clinical mastitis, milk and SCC test-day recordings, results of bacteriological cultures (Escherichia coli, Staphylococcus aureus, Streptococcus uberis, coagulase-negative staphylococci, Streptococcus dysgalactiae, and streptococci other than Strep. dysgalactiae and Strep. uberis), and cow characteristics. Following a diagnosis of clinical mastitis, the cow was treated and changes (increase or decrease) in milk production before and after a diagnosis were interpreted counterfactually. On a daily basis, indirect changes, mediated by SCC increase, were significantly different from zero for all bacterial species, with a milk yield decrease (ranging among species from 4 to 33g and mediated by an increase of 1000 SCC/mL/day) before and a daily milk increase (ranging among species from 2 to 12g and mediated by a decrease of 1000 SCC/mL/day) after detection. Direct changes, not mediated by SCC, were only different from zero for coagulase-negative staphylococci before diagnosis (72g per day). We concluded that mixed structural equation models were useful to estimate direct and indirect effects of the presence of clinical mastitis on milk yield. Copyright © 2015 Elsevier B.V. All rights reserved.

  9. Indirect serarches for very heavy quarks

    International Nuclear Information System (INIS)

    Bigi, I.I.

    1987-03-01

    Detailed studies of weak decays can reveal the presence of very massive quanta like heavy top quarks or fourth family quarks. The decay K + → π + ν anti ν and B/sub d/ - anti B/sub d/ mixing are particularly promising fields for such searches. We infer a rather conservative lower limit of 70 GeV on the top mass form recent ARGUS data on B/sub d/ - anti B/sub d/ mixing, near-maximal B 8 - anti B 8 mixing is another consequence. If on the other hand top were detected in Z 0 decays, then the presence of New Physics would be established in B 0 decays. The ratio between tau(B 0 ) and tau(B +- ) is of considerable phenomenological relevance here

  10. Indirect monitoring of internal exposure for actinides

    Energy Technology Data Exchange (ETDEWEB)

    Carneiro G, C.J.; Barreto F, J.; Todo A, A. [Instituto de Pesquisas Energeticas e Nucleares, IPEN/CNEN, Av. Professor Lineu Prestes No. 2242, Zip code 05508-000, Sao Paulo (Brazil)

    2006-07-01

    The procedure used to the assessment of internal exposure of workers involved with dismantling lightning rods and radioactive smoke detectors is described. Due to the presence of the sources of {sup 241} Am in these devices, a monitoring program to the workers have been implemented. This paper presents an analytical method for the separation and analysis of plutonium (Pu) and americium (Am) in urine samples using solid-phase extraction chromatography and alpha spectrometry. The mean recovery obtained with this technique is about 80% and the detection limit for 24 h urine sample range between 0.6 mBqL{sup -1} and 1.0 mBqL{sup -1}. The assessment of intakes and internal doses are performed following ICRP Publication 78 recommendations and appropriated biokinetic models (ICRP, 1997). Assumptions have been made for routine monitoring of these workers and it is also discussed the establishment of the internal monitoring program using the results of alpha measurements. (Author)

  11. Indirect monitoring of internal exposure for actinides

    International Nuclear Information System (INIS)

    Carneiro G, C.J.; Barreto F, J.; Todo A, A.

    2006-01-01

    The procedure used to the assessment of internal exposure of workers involved with dismantling lightning rods and radioactive smoke detectors is described. Due to the presence of the sources of 241 Am in these devices, a monitoring program to the workers have been implemented. This paper presents an analytical method for the separation and analysis of plutonium (Pu) and americium (Am) in urine samples using solid-phase extraction chromatography and alpha spectrometry. The mean recovery obtained with this technique is about 80% and the detection limit for 24 h urine sample range between 0.6 mBqL -1 and 1.0 mBqL -1 . The assessment of intakes and internal doses are performed following ICRP Publication 78 recommendations and appropriated biokinetic models (ICRP, 1997). Assumptions have been made for routine monitoring of these workers and it is also discussed the establishment of the internal monitoring program using the results of alpha measurements. (Author)

  12. Evaluation of a direct immunofluorescent antibody (difma test using Leishmania genus - specific monoclonal antibody in the routine diagnosis of cutaneous leishmaniasis

    Directory of Open Access Journals (Sweden)

    Martha E. Chico

    1995-06-01

    Full Text Available A direct immunofluorescent antibody (DIFMA test using a Leishmania genus- specific monoclonal antibody was evaluated in the routine diagnosis of cutaneous leishmaniasis (CL in Ecuador. This test was compared with the standard diagnostic techniques of scrapings, culture and histology. Diagnostic samples were taken from a total of 90 active dermal ulcers from patients from areas of Ecuador known to be endemic for cutaneous leishmaniasis. DIFMA was positive in all lesions. It was shown to be significantly superior to standard diagnostic methods either alone or in combination. The sensitivity of DIFMA did not diminish with chronicity of lesions. This test proved to be extremely useful in the routine diagnosis of CL because it is highly sensitive, is easy to use and produces rapid results.

  13. Modification of a whole room indirect calorimeter for measurement of rapid changes in energy expenditure.

    Science.gov (United States)

    Sun, M; Reed, G W; Hill, J O

    1994-06-01

    Whole room indirect calorimeters are among the most accurate devices for measurement of human energy expenditure and have provided useful data about determinants of total daily energy expenditure. However, a limitation of whole room indirect calorimeters has been the inability to detect acute (usually calorimeter (respiratory chamber) to allow accurate measurement of energy expenditure over time periods as short as 1 min. The modifications involve changes in the system design and use of signal processing techniques. With these modifications, we can measure energy expenditure in 1-min intervals throughout the day. This allows accurate study of the acute effects of food, exercise, or drugs on energy expenditure in subjects moving freely inside the respiratory chamber. The ability to use respiratory chambers for these types of studies should improve our understanding of how body weight is regulated.

  14. Utilizing optical coherence tomography for CAD/CAM of indirect dental restorations

    Science.gov (United States)

    Chityala, Ravishankar; Vidal, Carola; Jones, Robert

    Optical Coherence Tomography (OCT) has seen broad application in dentistry including early carious lesion detection and imaging defects in resin composite restorations. This study investigates expanding the clinical usefulness by investigating methods to use OCT for obtaining three-dimensional (3D) digital impressions, which can be integrated to CAD/CAM manufacturing of indirect restorations. 3D surface topography `before' and `after' a cavity preparation was acquired by an intraoral cross polarization swept source OCT (CP-OCT) system with a Micro-Electro-Mechanical System (MEMS) scanning mirror. Image registration and segmentation methods were used to digitally construct a replacement restoration that modeled the original surface morphology of a hydroxyapatite sample. After high resolution additive manufacturing (e.g. polymer 3D printing) of the replacement restoration, micro-CT imaging was performed to examine the marginal adaptation. This study establishes the protocol for further investigation of integrating OCT with CAD/CAM of indirect dental restorations.

  15. Indirect Inference for Stochastic Differential Equations Based on Moment Expansions

    KAUST Repository

    Ballesio, Marco; Tempone, Raul; Vilanova, Pedro

    2016-01-01

    We provide an indirect inference method to estimate the parameters of timehomogeneous scalar diffusion and jump diffusion processes. We obtain a system of ODEs that approximate the time evolution of the first two moments of the process

  16. An Indirect Assessment of the War Against Iraq

    National Research Council Canada - National Science Library

    Ferguson Jr, C. M

    1992-01-01

    ...? General Schwartzkopf's statement of intent reveals that his strategy for the conduct of operations reflected an appreciation for the "indirect approach," of Sun Tzu and his erstwhile protege, B. H. Liddell-Hart...

  17. Separating astrophysical sources from indirect dark matter signals

    Science.gov (United States)

    Siegal-Gaskins, Jennifer M.

    2015-01-01

    Indirect searches for products of dark matter annihilation and decay face the challenge of identifying an uncertain and subdominant signal in the presence of uncertain backgrounds. Two valuable approaches to this problem are (i) using analysis methods which take advantage of different features in the energy spectrum and angular distribution of the signal and backgrounds and (ii) more accurately characterizing backgrounds, which allows for more robust identification of possible signals. These two approaches are complementary and can be significantly strengthened when used together. I review the status of indirect searches with gamma rays using two promising targets, the Inner Galaxy and the isotropic gamma-ray background. For both targets, uncertainties in the properties of backgrounds are a major limitation to the sensitivity of indirect searches. I then highlight approaches which can enhance the sensitivity of indirect searches using these targets. PMID:25304638

  18. Indirect Monetary Policy Reforms and Output Growth in Nigeria: An ...

    African Journals Online (AJOL)

    BRRE). The efforts in repositioning these banks through the current banking reforms (recapitalization and consolidation) the paper notes are a right step in the right direction. Keywords: Indirect monetary policy, Granger – causality, output growth

  19. Indirect inference with time series observed with error

    DEFF Research Database (Denmark)

    Rossi, Eduardo; Santucci de Magistris, Paolo

    estimation. We propose to solve this inconsistency by jointly estimating the nuisance and the structural parameters. Under standard assumptions, this estimator is consistent and asymptotically normal. A condition for the identification of ARMA plus noise is obtained. The proposed methodology is used......We analyze the properties of the indirect inference estimator when the observed series are contaminated by measurement error. We show that the indirect inference estimates are asymptotically biased when the nuisance parameters of the measurement error distribution are neglected in the indirect...... to estimate the parameters of continuous-time stochastic volatility models with auxiliary specifications based on realized volatility measures. Monte Carlo simulations shows the bias reduction of the indirect estimates obtained when the microstructure noise is explicitly modeled. Finally, an empirical...

  20. RESEMBLANCE OF INDIRECTNESS IN POLITENESS OF EFL LEARNERS’ REQUEST REALIZATIONS

    Directory of Open Access Journals (Sweden)

    Indawan Syahri

    2013-07-01

    Full Text Available Abstract: Politeness principles are universally utilized by the speakers of any language when realizing various speech acts. However, the speakers of particular languages relatively apply politeness due to the cultural norms embedded. The present study attempts to delineate how the Indonesian learners of English (ILE apply the politeness principles in request realizations. Specifically it devotes to the types of politeness strategies applied and resemblance of the indirectness in politeness strategies in requesting acts. The FTAs and indirectness are the theoretical bases used to trace the typologies of both politeness and request strategies. The data werere collected by means of certain elicitation techniques, i.e. DCTs and Role-plays. The analyses werere done through three stages; determining request strategies, politeness strategies, and resemblance of indirectness in politeness. The results show that the indirectness generally is parallel to politeness. Besides, some pragmatic transfers are found in terms of applying native-culture norms in realizing target speech acts.