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Sample records for specific compound immunofluorescence

  1. Yellow fever monoclonal antibodies: type-specific and cross-reactive determinants identified by immunofluorescence.

    Science.gov (United States)

    Monath, T P; Schlesinger, J J; Brandriss, M W; Cropp, C B; Prange, W C

    1984-07-01

    Monoclonal antibodies directed against the envelope glycoprotein and the NV3 non-structural viral protein of yellow fever (YF) were tested by the indirect fluorescent antibody technique against a variety of YF virus strains and heterologous flaviviruses. Monoclonal antibodies directed against the envelope glycoprotein exhibited YF strain-specificity, YF type-specificity, broad group cross-reactivity, or limited subgroup reactivity (YF + Banzi or YF + Koutango + Zika + Usutu + Uganda S). Monoclonal antibodies directed against NV3 reacted either with YF + Koutango or with YF + Banzi. These findings generally correlated with the results of biological tests reported previously. Monoclonal antibodies that were type-specific to YF will be useful for the rapid specific identification of YF virus isolates and are available from the Centers for Disease Control on request.

  2. Immuno-fluorescence based Vi capsular polysaccharide detection for specific recognition of Salmonella enterica serovar Typhi in clinical samples.

    Science.gov (United States)

    Pandey, Satish K; Vinayaka, Aaydha C; Rishi, Dharam B; Rishi, Praveen; Suri, C Raman

    2014-09-02

    Typhoid fever is a life threatening bacterial infection that remains a major global health concern. This continued high burden associated with significant morbidity and mortality rate demands specific and rapid detection technique. This work reports a new sandwich type fluorescence immunoassay format using polymyxin B, a cationic receptor molecule, as a binder agent while anti-Vi antibody served as the capturing agent for specifically detecting Salmonella enterica serovar Typhi. Anti-Vi IgG antibody raised against Vi-BSA conjugate revealed affinity of 7.779nM(-1) signifying immunodominancy of O-acetyls groups in Vi polysaccharide. The detection limit of the developed assay was around 10(1) cellsmL(-1) of Vi expressing Salmonella enterica serovar Typhi with a correlation coefficient (R(2)) equal to 0.97. Positive response obtained for all the tested serovar Typhi clinical isolates as well as the pathogen spiked blood samples recommended specificity and accuracy of Vi antigen as a biomarker during typhoid fever. The intra- and inter-assay precision with Vi spiked samples were satisfactory revealing coefficient of variance (CV%) with a mean of 4.05% and 5.97% respectively. This may be the novel attempt and constructive report on the fluorescence based detection of Vi antigen of serovar Typhi in the epidemic as well as pandemic outbreaks. Copyright © 2014 Elsevier B.V. All rights reserved.

  3. 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.

  4. Stable expression of human muscle-specific kinase in HEp-2 M4 cells for automatic immunofluorescence diagnostics of myasthenia gravis.

    Science.gov (United States)

    George, Sandra; Paulick, Silvia; Knütter, Ilka; Röber, Nadja; Hiemann, Rico; Roggenbuck, Dirk; Conrad, Karsten; Küpper, Jan-Heiner

    2014-01-01

    Muscle-specific kinase (MuSK) belongs to the nicotinic acetylcholine receptor complex which is targeted by pathogenic autoantibodies causing Myasthenia gravis. While up to 95% of patients with generalized Myasthenia gravis were shown to be positive for acetylcholine receptor-specific autoantibodies, up to 70% of the remaining patients develop autoantibodies against MuSK. Discrimination of the autoantibody specificity is important for therapy of Myasthenia gravis. Recently, the new automatic fluorescence assessment platform AKLIDES has been developed for immunofluorescence-based diagnostics of autoimmune diseases. In order to establish an AKLIDES procedure for the detection of MuSK-specific autoantibodies (anti-MuSK), we developed a recombinant HEp-2 cell clone expressing the human MuSK cDNA. Here we show at the mRNA and protein level that the cell clone HEp-2 M4 stably expresses human MuSK. We provide evidence for a localization of MuSK at the cell membrane. Using cell clone HEp-2 M4 on the AKLIDES system, we investigated 34 patient sera that were previously tested anti-MuSK positive by radioimmunoassay as positive controls. As negative controls, we tested 29 acetylcholine receptor-positive but MuSK-negative patient sera, 30 amytrophic lateral sclerosis (ALS) patient sera and 45 blood donors. HEp-2 M4 cells revealed a high specificity for the detection of MuSK autoantibodies from 25 patient sera assessed by a specific pattern on HEp-2 M4 cells. By using appropriate cell culture additives, the fraction of cells stained positive with anti-MuSK containing sera can be increased from 2-16% to 10-48%, depending on the serum. In conclusion, we provide data showing that the novel recombinant cell line HEp-2 M4 can be used to screen for anti-MuSK with the automatic AKLIDES system.

  5. Stable expression of human muscle-specific kinase in HEp-2 M4 cells for automatic immunofluorescence diagnostics of myasthenia gravis.

    Directory of Open Access Journals (Sweden)

    Sandra George

    Full Text Available Muscle-specific kinase (MuSK belongs to the nicotinic acetylcholine receptor complex which is targeted by pathogenic autoantibodies causing Myasthenia gravis. While up to 95% of patients with generalized Myasthenia gravis were shown to be positive for acetylcholine receptor-specific autoantibodies, up to 70% of the remaining patients develop autoantibodies against MuSK. Discrimination of the autoantibody specificity is important for therapy of Myasthenia gravis. Recently, the new automatic fluorescence assessment platform AKLIDES has been developed for immunofluorescence-based diagnostics of autoimmune diseases. In order to establish an AKLIDES procedure for the detection of MuSK-specific autoantibodies (anti-MuSK, we developed a recombinant HEp-2 cell clone expressing the human MuSK cDNA. Here we show at the mRNA and protein level that the cell clone HEp-2 M4 stably expresses human MuSK. We provide evidence for a localization of MuSK at the cell membrane. Using cell clone HEp-2 M4 on the AKLIDES system, we investigated 34 patient sera that were previously tested anti-MuSK positive by radioimmunoassay as positive controls. As negative controls, we tested 29 acetylcholine receptor-positive but MuSK-negative patient sera, 30 amytrophic lateral sclerosis (ALS patient sera and 45 blood donors. HEp-2 M4 cells revealed a high specificity for the detection of MuSK autoantibodies from 25 patient sera assessed by a specific pattern on HEp-2 M4 cells. By using appropriate cell culture additives, the fraction of cells stained positive with anti-MuSK containing sera can be increased from 2-16% to 10-48%, depending on the serum. In conclusion, we provide data showing that the novel recombinant cell line HEp-2 M4 can be used to screen for anti-MuSK with the automatic AKLIDES system.

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

    Science.gov (United States)

    Qu, Yan-Gang; Zhang, Qian; Pan, Qi; Zhao, Xian-Da; Huang, Yan-Hua; Chen, Fu-Chun; Chen, Hong-Lei

    2014-01-01

    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). 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. 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; Pmutations (κ  =0.826; Pmutations with its high sensitivity and specificity, as compared with real-time polymerase chain reaction. In addition, the development of specific antibodies against EGFR mutation proteins might be useful for the diagnosis and treatment of lung cancer.

  7. 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.O método de imunofluorescência direta (DIFMA, com anticorpos monoclonais gênero- específicos para Leishmania, foi avaliado na rotina diagnostica da leishmaniose cutânea no Equador. O método foi comparado com técnicas diagnosticas de rotina: o esfregaço, a cultura e o exame histopatolôgico. As amostras para o diagnóstico foram obtidas de um total de 90 lesões cutâneas ativas, de doentes das ãreas do Equador, endêmicas para leishmaniose cutânea. O DIFMA foi positivo em todas as lesões, com resultados significativamente superior aos métodos diagnósticos de rotina, isolado ou em combinação. A sensibilidade do DFIMA não diminui em lesões crônicas. O método mostra-se muito útil no diagnóstico de leishmaniose cutânea, pela sua sensibilidade, rapidez e facilidade de execução.

  8. Immunofluorescence in dermatology

    Directory of Open Access Journals (Sweden)

    Seema Chhabra

    2012-01-01

    Full Text Available Direct immunofluorescence (DIF and indirect immunofluorescence (IIF tests on skin biopsy are being done mostly in academic teaching hospitals. These tests provide a useful diagnostic aid to dermatologists. Immunohistology and serology can, in conjunction with histology, provide considerable help in delineation and diagnosis of various skin disorders as well as systemic diseases with skin involvement, e.g. systemic lupus erythematosus. Immunofluorescence (IF studies have now become an invaluable supplement to clinical and histological examination in a variety of dermatological diseases. These skin diseases now include not only bullous and connective tissue disorders, vasculitides, and conditions such as lichen planus, but also the scaling dermatoses, notably psoriasis. In this review article, we share our experience of providing such a diagnostic facility for more than 30 years in a large tertiary care health center in North India and also help to outline the conditions, which can be diagnosed confidently, and others where IF can help in confirming a diagnosis or the immune component of the disease. The article also deals with handling of skin biopsy specimens and interpretation of biopsy findings on DIF and IIF examination.

  9. Specificity of granular IgM deposition in folliculosebaceous units and sweat gland apparatus in direct immunofluorescence (DIF) of lupus erythematosus.

    Science.gov (United States)

    Elbendary, Amira; Zhou, Cheng; Valdebran, Manuel; Yu, Yan; Gad, AbdAllah; Kwon, Eun Ji; Elston, Dirk M

    2016-08-01

    Diagnosis of lupus erythematosus (LE) in direct immunofluorescence testing is based on the finding of positive immunofluorescence at the dermoepidermal junction (DEJ). We sought to evaluate the sensitivity of IgM deposition at the DEJ and adnexal structures in the diagnosis of lupus erythematosus. We conducted a retrospective study of 100 previously diagnosed cases of lupus erythematosus and 158 cases of other immune-mediated dermatosis. Deposition of IgG, IgM, IgA, and C3 at the DEJ, follicular units, and sweat glands were recorded. Presence or absence of adnexal structures was documented. The immunoreactant deposition was documented as linear, coarse granular, or stippled. The most frequently deposited immunoreactant in lupus erythematosus cases was IgM along the DEJ and stromal-epithelial junction of hair follicles and sweat glands. IgM deposition along the stromal-epithelial junction of hair follicles and sweat glands was strongly associated with a diagnosis of lupus erythematosus compared with other immune-mediated diseases collectively (P value < .001). The pattern of IgM in lupus and dermatomyositis is granular, in contrast to the linear deposition in the other disorders evaluated. This was a retrospective study of archived material. Granular IgM deposition at the stromal-epithelial junction of cutaneous adnexal structures suggests a diagnosis of lupus erythematosus or dermatomyositis. Copyright © 2016 American Academy of Dermatology, Inc. All rights reserved.

  10. Immunofluorescence of Autoimmune Bullous Diseases

    NARCIS (Netherlands)

    Diercks, Gilles F; Pas, Hendri H; Jonkman, Marcel F

    Autoimmmune bullous diseases of skin and mucosa are uncommon, disabling, and potentially lethal diseases. For a quick and reliable diagnosis immunofluorescence is essential. This article describes two variants of immunofluorescence. The direct method uses a skin or mucosal biopsy of the patient to

  11. Structuring of bacterioplankton communities by specific dissolved organic carbon compounds.

    Science.gov (United States)

    Gómez-Consarnau, Laura; Lindh, Markus V; Gasol, Josep M; Pinhassi, Jarone

    2012-09-01

    The main role of microorganisms in the cycling of the bulk dissolved organic carbon pool in the ocean is well established. Nevertheless, it remains unclear if particular bacteria preferentially utilize specific carbon compounds and whether such compounds have the potential to shape bacterial community composition. Enrichment experiments in the Mediterranean Sea, Baltic Sea and the North Sea (Skagerrak) showed that different low-molecular-weight organic compounds, with a proven importance for the growth of marine bacteria (e.g. amino acids, glucose, dimethylsulphoniopropionate, acetate or pyruvate), in most cases differentially stimulated bacterial growth. Denaturing gradient gel electrophoresis 'fingerprints' and 16S rRNA gene sequencing revealed that some bacterial phylotypes that became abundant were highly specific to enrichment with specific carbon compounds (e.g. Acinetobacter sp. B1-A3 with acetate or Psychromonas sp. B3-U1 with glucose). In contrast, other phylotypes increased in relative abundance in response to enrichment with several, or all, of the investigated carbon compounds (e.g. Neptuniibacter sp. M2-A4 with acetate, pyruvate and dimethylsulphoniopropionate, and Thalassobacter sp. M3-A3 with pyruvate and amino acids). Furthermore, different carbon compounds triggered the development of unique combinations of dominant phylotypes in several of the experiments. These results suggest that bacteria differ substantially in their abilities to utilize specific carbon compounds, with some bacteria being specialists and others having a more generalist strategy. Thus, changes in the supply or composition of the dissolved organic carbon pool can act as selective forces structuring bacterioplankton communities. © 2012 Society for Applied Microbiology and Blackwell Publishing Ltd.

  12. Production of N-13 labeled compounds with high specific activity

    Energy Technology Data Exchange (ETDEWEB)

    Suzuki, Kazutoshi; Sasaki, Motoji; Yoshida, Yuichiro; Haradahira, Terushi; Inoue, Osamu [National Inst. of Radiological Sciences, Chiba (Japan)

    1997-03-01

    Nitrogen-13 was produced by irradiating ultra pure water saturated with a pure gas (N2, O2, He, H2) with 18 MeV protons. Ion species generated by irradiation were analyzed with radio ion chromatography systems. An automated equipment was developed to synthesize anhydrous (13N)NH3 as a synthetic precursor and (13N)p-nitrophenyl carbamate ((13N)NPC) as a model compound, using the (13N)NH3. The radiochemical yield and specific activity of (13N)NPC was high enough to carry out the receptor study with PET. (author)

  13. Time-trends in method-specific suicide rates compared with the availability of specific compounds

    DEFF Research Database (Denmark)

    Nordentoft, Merete; Qin, Ping; Helweg-Larsen, Karin

    2006-01-01

    Restriction of means for suicide is an important part of suicide preventive strategies in different countries. All suicides in Denmark between 1970 and 2000 were examined with regard to method used for suicide. Overall suicide mortality and method-specific suicide mortality was compared...... in the number of suicides by self-poisoning with these compounds. Restricted access occurred concomittantly with a 55% decrease in suicide rate....

  14. Direct immunofluorescence for the diagnosis of legionellosis

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    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.

  15. Pharmacological promiscuity: dependence on compound properties and target specificity in a set of recent Roche compounds.

    Science.gov (United States)

    Peters, Jens-Uwe; Schnider, Patrick; Mattei, Patrizio; Kansy, Manfred

    2009-04-01

    The term "pharmacological promiscuity" describes a compound's pharmacological activity at multiple targets. Pharmacological promiscuity is undesired in typical drug discovery projects, which focus on the "one drug-one target" paradigm. Off-target activity can lead to adverse drug reactions, or can obscure pharmacodynamic effects in animal models. Therefore, advanced lead compounds, pharmacological tool compounds, and drug candidates are usually screened against panels of safety-relevant targets to detect unwanted pharmacological activities. To identify determinants of pharmacological promiscuity, we compared the panel screening outcomes of 213 recent Roche compounds with their molecular properties. Pronounced promiscuity was not observed below a threshold Clog P value of 2. For basic compounds, the propensity for weak off-target activity was found to increase with calculated basicities, whereas the potential for strong off-target activity depends more qualitatively on the presence of a positive charge at physiological pH. Compounds originating from projects with an aminergic receptor or transporter as a therapeutic target are particularly prone to promiscuity; the promiscuity of such compounds is mainly caused by their activity at other aminergic targets in the screening panel.

  16. Tracing carbon fixation in phytoplankton—compound specific and total

    NARCIS (Netherlands)

    Grosse, J.; Van Breugel, P.; Boschker, H.T.S.

    2015-01-01

    Measurement of total primary production using 13C incorporation is a widely established tool. However, these bulk measurements lack information about the fate of fixed carbon: the production of major cellular compounds (carbohydrates, amino acids, fatty acids, and DNA/RNA) is affected by for

  17. Nitrite and nitroso compounds can serve as specific catalase inhibitors.

    Science.gov (United States)

    Titov, Vladimir Yu; Osipov, Anatoly N

    2017-03-01

    We present evidence that nitrite and nitrosothiols, nitrosoamines and non-heme dinitrosyl iron complexes can reversibly inhibit catalase with equal effectiveness. Catalase activity was evaluated by the permanganatometric and calorimetric assays. This inhibition is not the result of chemical transformations of these compounds to a single inhibitor, as well as it is not the result of NO release from these substances (as NO traps have no effect on the extent of inhibition). It was found that chloride and bromide in concentration above 80 mM and thiocyanate in concentration above 20 μM enhance catalase inhibition by nitrite and the nitroso compounds more than 100 times. The inhibition degree in this case is comparable with that induced by azide. We propose that the direct catalase inhibitor is a positively charged NO-group. This group acquires a positive charge in the active center of enzyme by interaction of nitrite or nitroso compounds with some enzyme groups. Halides and thiocyanate protect the NO+ group from hydration and thus increase its inhibition effect. It is probable that a comparatively low chloride concentration in many cells is the main factor to protect catalase from inhibition by nitrite and nitroso compounds.

  18. Immunofluorescence antigen mapping for hereditary epidermolysis bullosa

    Directory of Open Access Journals (Sweden)

    Raghavendra Rao

    2012-01-01

    Full Text Available Epidermolysis bullosa (EB is a group of inherited, mechanobullous disorders that are caused by mutations in the structural proteins in the epidermis or dermoepidermal junction. Characteristic clinical picture is the presence of blisters at trauma prone areas of the body, which develops at or soon after birth. Availability of specific monoclonal antibodies against the target proteins together with advances in the molecular genetics have led to the revision in the classification of EB. Now four major types of EB are recognized depending upon the level of blister and the location of target protein: EB simplex (epidermolytic, junctional EB (lucidolytic, dystrophic EB (dermolytic and Kindler′s syndrome (mixed cleavage plane. The laboratory tests not only help to confirm the diagnosis of EB but are also an important tool to classify (and subtype EB. These include immunofluorescence antigen mapping (IFM, transmission electron microscopy (TEM and mutation analysis. IFM is the most preferred method for final diagnosis of EB worldwide. It is relatively easy to perform and results can be obtained rapidly. This article describes the technicalities and significance of IFM in various types of EB.

  19. [Immunofluorescence study of the extracellular matrix of the human placenta].

    Science.gov (United States)

    Rukosuev, V S; Fokin, E I; Milovanov, A P

    1989-01-01

    Distribution of collagen types I, III, IV, V and fibronectin in human placental villi has been studied by indirect immunofluorescence. During 9-12 weeks of pregnancy the extracellular matrix of villi represents a network of filaments organized in bundles and aggregates that contain collagen types I and III and finer filaments of collagen types IV and V. Collagen type IV is regularly detected in basal membrane of capillaries and particularly in villous epithelium, collagen type V and fibronectin are occasionally detected there. Marked immunofluorescent reaction on collagen types IV and V and fibronectin, and weak reaction on collagen type III is observed in cellular islets around cytotrophoblasts. In the fetus born in term placental villi have uniform immunofluorescence in thick basal membranes of fetal capillaries and of chorionic epithelium. The immunofluorescent reaction specific for all collagen types is uniform in villous stroma. Distribution of different collagen types and fibronectin, including the unusual localization of membrane collagen type IV, in villous stroma and cellular islets of early and mature placenta is discussed.

  20. Role of direct immunofluorescence in the diagnosis of glomerulonephritis

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    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.

  1. Compound-Specific Sulfur Isotope Analysis of Petroleum Gases.

    Science.gov (United States)

    Said-Ahmad, Ward; Wong, Kenneth; Mcnall, Monaca; Shawar, Lubna; Jacksier, Tracey; Turich, Courtney; Stankiewicz, Artur; Amrani, Alon

    2017-03-07

    We describe a simple, sensitive, and robust method for sulfur isotope ratio (34S/32S) analysis of ppm-level organic sulfur compounds (OSCs) in the presence of percent-level H2S. The method uses a gas chromatograph (GC) coupled with a multicollector inductively coupled plasma mass spectrometer (MC-ICPMS). The GC, equipped with a gas inlet and a valve that transfers the H2S to a thermal conductivity detector (TCD), enables a precise heart cut and prevents the saturation of the MC-ICPMS. The sensitivity and accuracy of the method are better than 0.3‰ for OSCs at a concentration of 25 pmol or 1.4 ppm, and better than 0.5‰ for concentrations ≥0.7 ppm of OSCs. An order of magnitude increase in sensitivity, with no effect on accuracy, can be achieved if the loop volume (0.5 mL) is changed to 5 mL. High concentrations of methane (95% v/v) and/or H2S (20% v/v) had no effect (within 0.5‰) on the precision and accuracy of the gas sample containing 2 ppm of OSCs after heart cut. The applicability and robustness of this method are demonstrated on a gas sample (10% v/v H2S) that was produced by pyrolysis of sulfur-rich kerogen. The results show good precision and reveal sulfur isotope variability between individual OSCs that may represent key processes during formation and degradation of OSCs.

  2. Compound

    Indian Academy of Sciences (India)

    UV-vis spectra showing solvent effects on compounds (6). Figure S4. UV-vis spectra showing solvent effects on compounds (9). Figure S5. UV-vis spectra showing solvent ___, acidic--- and basic -□- effects on compound (8) in CH2Cl2 solution. Table S1. 1H and 13C NMR spectral data of salicylaldimine Schiff bases (5-8).

  3. Time-trends in method-specific suicide rates compared with the availability of specific compounds. The Danish experience

    DEFF Research Database (Denmark)

    Nordentoft, Merete; Qin, Ping; Helweg-Larsen, Karin

    2006-01-01

    Restriction of means for suicide is an important part of suicide preventive strategies in different countries. All suicides in Denmark between 1970 and 2000 were examined with regard to method used for suicide. Overall suicide mortality and method-specific suicide mortality was compared...... in the number of suicides by self-poisoning with these compounds. Restricted access occurred concomittantly with a 55% decrease in suicide rate...

  4. Screening in larval zebrafish reveals tissue-specific distribution of fifteen fluorescent compounds

    Directory of Open Access Journals (Sweden)

    Yuxiao Yao

    2017-09-01

    Full Text Available The zebrafish is a prominent vertebrate model for low-cost in vivo whole organism screening. In our recent screening of the distribution patterns of fluorescent compounds in live zebrafish larvae, fifteen compounds with tissue-specific distributions were identified. Several compounds were observed to accumulate in tissues where they were reported to induce side-effects, and compounds with similar structures tended to be enriched in the same tissues, with minor differences. In particular, we found three novel red fluorescent bone-staining dyes: purpurin, lucidin and 3-hydroxy-morindone; purpurin can effectively label bones in both larval and adult zebrafish, as well as in postnatal mice, without significantly affecting bone mass and density. Moreover, two structurally similar chemotherapeutic compounds, doxorubicin and epirubicin, were observed to have distinct distribution preferences in zebrafish. Epirubicin maintained a relatively higher concentration in the liver, and performed better in inhibiting hepatic hyperplasia caused by the over-expression of krasG12V. In total, our study suggests that the transparent zebrafish larvae serve as valuable tools for identifying tissue-specific distributions of fluorescent compounds.

  5. Degradation of specific aromatic compounds migrating from PEX pipes into drinking water.

    Science.gov (United States)

    Ryssel, Sune Thyge; Arvin, Erik; Lützhøft, Hans-Christian Holten; Olsson, Mikael Emil; Procházková, Zuzana; Albrechtsen, Hans-Jørgen

    2015-09-15

    Nine specific compounds identified to migrate from polyethylene (PE) and cross-linked polyethylene (PEX) to drinking water were investigated for their degradation in drinking water. Three sample types were studied: field samples (collected at consumer taps), PEX pipe water extractions, and water samples spiked with target compounds. Four compounds were quantified in field samples at concentrations of 0.15-8.0 μg/L. During PEX pipe water extraction 0.42 ± 0.20 mg NVOC/L was released and five compounds quantified (0.5-6.1 μg/L). The degradation of these compounds was evaluated in PEX-pipe water extractions and spiked samples. 4-ethylphenol was degraded within 22 days. Eight compounds were, however, only partially degradable under abiotic and biotic conditions within the timeframe of the experiments (2-4 weeks). Neither inhibition nor co-metabolism was observed in the presence of acetate or PEX pipe derived NVOC. Furthermore, the degradation in drinking water from four different locations with three different water works was similar. In conclusion, eight out of the nine compounds studied would - if being released from the pipes - reach consumers with only minor concentration decrease during water distribution. Copyright © 2015 Elsevier Ltd. All rights reserved.

  6. Role of direct immunofluorescence in dermatological disorders

    Directory of Open Access Journals (Sweden)

    Vijaya V Mysorekar

    2015-01-01

    Full Text Available Background: Direct immunofluorescence (DIF test for tissue-bound autoantibodies, has been found to be of value in the diagnosis of several dermatological disorders. The location and pattern of deposition of immunoreactants helps in classifying various immune-mediated diseases. Aims and Objectives: The aim of this study was to analyze the concordance between the clinical, histopathological and DIF diagnosis in bullous and nonbullous lesions of the skin, and thus determine the impact of immunofluorescence on diagnosis. Materials and Methods: A total of 215 skin biopsies performed in suspected immune-mediated vesiculobullous disease, vasculitis or dermatosis, were studied. Histopathological examination was done along with DIF study for deposits of immunoglobulin G(IgG, IgA, IgM, and C3. Results: Direct immunofluorescence was positive in 103/215 cases. There was very good concordance between the clinical, histological and DIF results (observed agreement = 93.4%, κ =0.90, with 95% confidence interval = 0.86-0.94. The overall sensitivity of DIF in immune-mediated skin disorders was 98.0%. DIF was positive in 52/53 cases (98.1% in the pemphigus group and 24/25 (96.0% bullous pemphigoid cases. None of the clinically suspected cases of dermatitis herpetiformis showed DIF positivity. A positive lupus band test was seen in 9/9 (100% cases of lupus erythematosus. DIF was positive in 10/10 (100% clinically suspected cases of Henoch-Schönlein purpura. In 110 cases, negative DIF results helped to rule out immune-mediated vesiculobullous disorders, lupus erythematosus and vasculitis, and the final diagnosis was made on the basis of the clinical features and/or histopathology. Conclusion: Direct immunofluorescence is a useful supplement for the accurate diagnosis of immune-mediated dermatological disorders, and helps to classify various autoimmune bullous disorders. When the clinical features/histopathology are inconclusive, the diagnosis often can be made on

  7. 21 CFR 866.3370 - Mycobacterium tuberculosis immunofluorescent reagents.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Mycobacterium tuberculosis immunofluorescent... § 866.3370 Mycobacterium tuberculosis immunofluorescent reagents. (a) Identification. Mycobacterium... used to identify Mycobacterium tuberculosis directly from clinical specimens. The identification aids...

  8. Low temperature specific heat of the spin-density-wave compound (TMTSF)2PF6

    DEFF Research Database (Denmark)

    Odin, J.; Lasjaunias, J.C.; Biljakovic, K.

    1994-01-01

    We report on specific heat measurements of the SDW compound (TMTSF)2PF6 between 2 and 25 K, performed by two different techniques. We discuss the two successive transitions which occur in this T-range : the SDW ordering transition at T = 12.1 K, and a glass transition around-3-3.5 K. The latter...

  9. 21 CFR 866.3460 - Rabiesvirus immuno-fluorescent reagents.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Rabiesvirus immuno-fluorescent reagents. 866.3460... immuno-fluorescent reagents. (a) Identification. Rabiesvirus immunofluorescent reagents are devices that consist of rabiesvirus antisera conjugated with a fluorescent dye used to identify rabiesvirus in...

  10. 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

  11. Proof of the quantitative potential of immunofluorescence by mass spectrometry.

    Science.gov (United States)

    Toki, Maria I; Cecchi, Fabiola; Hembrough, Todd; Syrigos, Konstantinos N; Rimm, David L

    2017-03-01

    Protein expression in formalin-fixed, paraffin-embedded patient tissue is routinely measured by Immunohistochemistry (IHC). However, IHC has been shown to be subject to variability in sensitivity, specificity and reproducibility, and is generally, at best, considered semi-quantitative. Mass spectrometry (MS) is considered by many to be the criterion standard for protein measurement, offering high sensitivity, specificity, and objective molecular quantification. Here, we seek to show that quantitative immunofluorescence (QIF) with standardization can achieve quantitative results comparable to MS. Epidermal growth factor receptor (EGFR) was measured by quantitative immunofluorescence in 15 cell lines with a wide range of EGFR expression, using different primary antibody concentrations, including the optimal signal-to-noise concentration after quantitative titration. QIF target measurement was then compared to the absolute EGFR concentration measured by Liquid Tissue-selected reaction monitoring mass spectrometry. The best agreement between the two assays was found when the EGFR primary antibody was used at the optimal signal-to-noise concentration, revealing a strong linear regression (R 2 =0.88). This demonstrates that quantitative optimization of titration by calculation of signal-to-noise ratio allows QIF to be standardized to MS and can therefore be used to assess absolute protein concentration in a linear and reproducible manner.

  12. Direct compound-specific stable chlorine isotope analysis of organic compounds with quadrupole GC/MS using standard isotope bracketing.

    Science.gov (United States)

    Aeppli, Christoph; Holmstrand, Henry; Andersson, Per; Gustafsson, Orjan

    2010-01-01

    A method has been developed for the direct determination of the stable chlorine isotope composition (delta(37)Cl) of organochlorines that eliminates sample preparation, achieves precision comparable to earlier techniques while improving the sensitivity, and makes use of benchtop gas chromatography-quadrupole mass spectrometry instruments (GCqMS). The method is based on the use of multiple injections (n = 8-10) of the sample, bracketed by a molecularly identical isotopic standard with known delta(37)Cl, determined using off-line thermal ionization mass spectrometry (TIMS). Mass traces of two isotopologues differing by one chlorine isotope were used to calculate delta(37)Cl values. Optimization of mass spectrometry and peak integration parameters as well as method validation was achieved using tetrachloroethene (PCE), p,p'-dichlorodiphenyltrichloroethane (DDT), and pentachlorophenol (PCP), spanning a delta(37)Cl range of -5.5 to +3.2 per thousand vs SMOC. Injecting 1.6-1100 pmol resulted in standard deviations (1sigma) of 0.6-1.3 per thousand, and the delta(37)Cl results agreed with values independently measured with TIMS. The method was tested by determining the Rayleigh fractionation during evaporation of pure liquid PCE, resulting in a chlorine isotopic enrichment factor of epsilon(Cl) = -1.1 +/- 0.4 per thousand. Furthermore, position-specific delta(37)Cl analysis based on analysis of DDT mass fragments was evaluated. The GCqMS-delta(37)Cl method offers a simplified yet sensitive approach for compound-specific chlorine isotope analysis.

  13. Multi-element compound specific stable isotope analysis of volatile organic compounds at trace levels in groundwater samples

    Science.gov (United States)

    Herrero-Martín, Sara; Nijenhuis, Ivonne; Schmidt, Marie; Wolfram, Diana; Richnow, Hans. H.; Gehre, Matthias

    2013-04-01

    Groundwater pollution remains one of the major environmental and health concerns. A thorough understanding of sources, sinks and transformation processes of groundwater contaminants is needed to improve risk management evaluation, and to design efficient remediation and water treatment strategies. Isotopic tools provide unique information for an in-depth understanding of the fate of organic chemicals in the environment. During the last decades compound specific isotope analysis (CSIA) of complex mixtures, using gas chromatography-isotope ratio mass spectrometry (GC-IRMS), has gained popularity for the characterization and risk assessment of hazardous waste sites and for isotope forensics of organic contaminants. Multi-element isotope fingerprinting of organic substances provides a more robust framework for interpretation than the isotope analysis of only one element. One major challenge for application of CSIA is the analysis of trace levels of organic compounds in environmental matrices. It is necessary to inject 1 nmol carbon or 8 nmol hydrogen on column, to obtain an accurate and precise measurement of the isotope ratios, which is between two and three orders of magnitude larger than the amount of compound needed for conventional analysis of compound concentrations. Therefore, efficient extraction and pre-concentration techniques have to be integrated with GC-IRMS. Further research is urgently needed in this field, to evaluate the potential of novel and environmental-friendly sample pre-treatment techniques for CSIA to lower the detection limits and extending environmental applications. In this study, the novel coupling of a headspace autosampler (HS) with a programmed temperature vaporizer (PTV), allowing large volume injection of headspace samples, is proposed to improve the sensitivity of CSIA. This automatic, fast and solvent free strategy provides a significant increase on the sensitivity of GC-based methods maintaining the simple headspace instrumentation

  14. Optimization of the solvent-based dissolution method to sample volatile organic compound vapors for compound-specific isotope analysis.

    Science.gov (United States)

    Bouchard, Daniel; Wanner, Philipp; Luo, Hong; McLoughlin, Patrick W; Henderson, James K; Pirkle, Robert J; Hunkeler, Daniel

    2017-10-20

    The methodology of the solvent-based dissolution method used to sample gas phase volatile organic compounds (VOC) for compound-specific isotope analysis (CSIA) was optimized to lower the method detection limits for TCE and benzene. The sampling methodology previously evaluated by [1] consists in pulling the air through a solvent to dissolve and accumulate the gaseous VOC. After the sampling process, the solvent can then be treated similarly as groundwater samples to perform routine CSIA by diluting an aliquot of the solvent into water to reach the required concentration of the targeted contaminant. Among solvents tested, tetraethylene glycol dimethyl ether (TGDE) showed the best aptitude for the method. TGDE has a great affinity with TCE and benzene, hence efficiently dissolving the compounds during their transition through the solvent. The method detection limit for TCE (5±1μg/m3) and benzene (1.7±0.5μg/m3) is lower when using TGDE compared to methanol, which was previously used (385μg/m3 for TCE and 130μg/m3 for benzene) [2]. The method detection limit refers to the minimal gas phase concentration in ambient air required to load sufficient VOC mass into TGDE to perform δ13C analysis. Due to a different analytical procedure, the method detection limit associated with δ37Cl analysis was found to be 156±6μg/m3 for TCE. Furthermore, the experimental results validated the relationship between the gas phase TCE and the progressive accumulation of dissolved TCE in the solvent during the sampling process. Accordingly, based on the air-solvent partitioning coefficient, the sampling methodology (e.g. sampling rate, sampling duration, amount of solvent) and the final TCE concentration in the solvent, the concentration of TCE in the gas phase prevailing during the sampling event can be determined. Moreover, the possibility to analyse for TCE concentration in the solvent after sampling (or other targeted VOCs) allows the field deployment of the sampling method

  15. Identification of fluorescent compounds with non-specific binding property via high throughput live cell microscopy.

    Directory of Open Access Journals (Sweden)

    Sangeeta Nath

    Full Text Available INTRODUCTION: Compounds exhibiting low non-specific intracellular binding or non-stickiness are concomitant with rapid clearing and in high demand for live-cell imaging assays because they allow for intracellular receptor localization with a high signal/noise ratio. The non-stickiness property is particularly important for imaging intracellular receptors due to the equilibria involved. METHOD: Three mammalian cell lines with diverse genetic backgrounds were used to screen a combinatorial fluorescence library via high throughput live cell microscopy for potential ligands with high in- and out-flux properties. The binding properties of ligands identified from the first screen were subsequently validated on plant root hair. A correlative analysis was then performed between each ligand and its corresponding physiochemical and structural properties. RESULTS: The non-stickiness property of each ligand was quantified as a function of the temporal uptake and retention on a cell-by-cell basis. Our data shows that (i mammalian systems can serve as a pre-screening tool for complex plant species that are not amenable to high-throughput imaging; (ii retention and spatial localization of chemical compounds vary within and between each cell line; and (iii the structural similarities of compounds can infer their non-specific binding properties. CONCLUSION: We have validated a protocol for identifying chemical compounds with non-specific binding properties that is testable across diverse species. Further analysis reveals an overlap between the non-stickiness property and the structural similarity of compounds. The net result is a more robust screening assay for identifying desirable ligands that can be used to monitor intracellular localization. Several new applications of the screening protocol and results are also presented.

  16. Evaluation of functional groups responsible for chloroform formation during water chlorination using compound specific isotope analysis.

    Science.gov (United States)

    Arnold, William A; Bolotin, Jakov; Von Gunten, Urs; Hofstetter, Thomas B

    2008-11-01

    Compound-specific isotope analysis was used to monitor the delta(13)C signature of chloroform produced upon the chlorination of model compounds representing natural organic matter functional groups (resorcinol, acetylacetone, acetophenone, phenol, and 2,4,6-trichlorophenol) and a natural water sample. For each model compound, a different apparent kinetic isotope effect was found for chloroform formation. Normal isotope effects were found for resorcinol, acetylacetone, and acetophenone, and ranged from 1.009 +/- 0.002 to 1.024 +/- 0.004. For the two phenols, an inverse effect was found (0.980 +/- 0.004). Lake Zürich water also had a inverse effect (0.997 +/- NOM, but that other functional groups may also participate. The apparent 13C kinetic isotope effect for the addition/ elimination reaction of 1,1,1-trichloropropanone mediated by OH- to yield chloroform is 1.014 +/- 0.002. A comparison of this value to those found for the chlorination of the model precursors and an evaluation of the differences in chloroform production kinetics for the different model precursors argue against a mechanism in which all NOM precursors react via a common intermediate. Compound specific isotope analysis may give additional insights into chloroform formation mechanisms beyond those allowed by current techniques.

  17. Grass material as process standard for compound-specific radiocarbon analysis

    Science.gov (United States)

    Cisneros-Dozal, Malu; Xu, Xiaomei; Bryant, Charlotte; Pearson, Emma; Dungait, Jennifer

    2015-04-01

    Compound-specific radiocarbon analysis (CSRA) is a powerful tool to study the carbon cycle and/or as a dating technique in paleoclimate reconstructions. The radiocarbon value of individual compounds can provide insight into turnover times, organic matter sources and in specific cases can be used to establish chronologies when traditional dating materials (e.g. macrofossils, pollen, charcoal) are not available. The isolation of compounds (or group of compounds) from parent material (e.g. soil, plant) for radiocarbon analysis can, however, introduce carbon contamination through chemical separation steps and preparative capillary gas chromatography (PCGC). In addition, the compounds of interest are often in low abundance which amplifies the contamination effect. The extraneous carbon can be of modern 14C age and/or 14C -free and its amount and 14C value must be determined for a given system/isolation procedure in order to report accurate 14C values. This can be achieved by using adequate standard materials but, by contrast with traditional radiocarbon dating, there are not established reference standards for CSRA work, in part because the type of standard material depends on the compounds of interest and the isolation procedure. Here we evaluate the use of n-alkanes extracted from single-year growth grass as modern process standard material for CSRA using PCGC isolation. The grass material has a known 14C value of 1.224 ± 0.006 fraction modern (FM) and the individual n-alkanes are expected to have a similar 14C value. In order to correct for the addition of extraneous carbon during PCGC isolation of the n-alkanes, we used commercially available compounds of modern 14C content and 14C -free (adipic acid, FM= 0.0015 ± 0.0001 and docosane, FM=1.059 ± 0.003) to evaluate our PCGC procedure. The corrected 14C values of the isolated n-alkanes extracted from the modern grass are within one sigma of the grass bulk 14C value for n-C29 and within two sigma for n-C23-C27, C31

  18. Carbon sources in the Beaufort Sea revealed by molecular lipid biomarkers and compound specific isotope analysis

    OpenAIRE

    Tolosa, Imma; Fiorini, S.; Gasser, Beat; Martín, Jacobo; Miquel, J.C.

    2012-01-01

    Molecular lipid biomarkers (hydrocarbons, alcohols, sterols and fatty acids) and compound specific isotope analysis of suspended particulate organic matter (SPM) and surface sediments of the Mackenzie Shelf and slope (Southeast Beaufort Sea, Arctic Ocean), were studied in summer 2009. The concentrations of the molecular lipid markers, characteristic of known organic matter sources, were grouped and used as proxies to evaluate the relative importance of fresh algal, detrital algal, foss...

  19. Performance specifications for technology development: Application for characterization of volatile organic compounds in the environment

    Energy Technology Data Exchange (ETDEWEB)

    Carpenter, S.E.; Doskey, P.V.; Erickson, M.D.; Lindahl, P.C.

    1994-07-01

    This report contains information about technology development for the monitoring and remediation of environmental pollution caused by the release of volatile organic compounds. Topics discussed include: performance specification processes, gas chromatography, mass spectrometer, fiber-optic chemical sensors, infrared spectroscopy, Raman spectroscopy, piezoelectric sensors and electrochemical sensors. These methods are analyzed for their cost efficiency, accuracy, and the ability to meet the needs of the customer.

  20. In Situ and Laboratory Studies on the Fate of Specific Organic Compounds in an Anerobic Landfill Leachate Plume, 2. Fate of Aromatic and Chlorinated Aliphatic Compounds

    DEFF Research Database (Denmark)

    Nielsen, Per Henning; Bjarnadóttir, Helga; Winter, Pia L.

    1995-01-01

    and laboratory batch microcosm experiments performed focusing on redox conditions, microbiology and the fate of 7 phenolic compounds. In this paper we present the results on the fate of 8 aromatic compounds and 4 chlorinated aliphatic compounds. Nitrobenzene was transformed at all distances from the landfill......The transformation of specific organic compounds was investigated by in situ and laboratory experiments in an anaerobic landfill leachate pollution plume at four different distances from the landfill. In a previous paper (Part 1, also published in this issue) we described the in situ microcosm...... the landfill. In general, good accordance with respect to compound transformation was observed between in situ and laboratory experiments, but in a few cases more compounds were transformed in in situ experiments than in the corresponding laboratory experiments....

  1. Novel HTS strategy identifies TRAIL-sensitizing compounds acting specifically through the caspase-8 apoptotic axis.

    Directory of Open Access Journals (Sweden)

    Darren Finlay

    Full Text Available Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand (TRAIL is potentially a very important therapeutic as it shows selectivity for inducing apoptosis in cancer cells whilst normal cells are refractory. TRAIL binding to its cognate receptors, Death Receptors-4 and -5, leads to recruitment of caspase-8 and classical activation of downstream effector caspases, leading to apoptosis. As with many drugs however, TRAIL's usefulness is limited by resistance, either innate or acquired. We describe here the development of a novel 384-well high-throughput screening (HTS strategy for identifying potential TRAIL-sensitizing agents that act solely in a caspase-8 dependent manner. By utilizing a TRAIL resistant cell line lacking caspase-8 (NB7 compared to the same cells reconstituted with the wild-type protein, or with a catalytically inactive point mutant of caspase-8, we are able to identify compounds that act specifically through the caspase-8 axis, rather than through general toxicity. In addition, false positive hits can easily be "weeded out" in this assay due to their activity in cells lacking caspase-8-inducible activity. Screening of the library of pharmacologically active compounds (LOPAC was performed as both proof-of-concept and to discover potential unknown TRAIL sensitizers whose mechanism is caspase-8 mediated. We identified known TRAIL sensitizers from the library and identified new compounds that appear to sensitize specifically through caspase-8. In sum, we demonstrate proof-of-concept and discovery of novel compounds with a screening strategy optimized for the detection of caspase-8 pathway-specific TRAIL sensitizers. This screen was performed in the 384-well format, but could easily be further miniaturized, allows easy identification of artifactual false positives, and is highly scalable to accommodate diverse libraries.

  2. An Automatic Indirect Immunofluorescence Cell Segmentation System

    Directory of Open Access Journals (Sweden)

    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.

  3. Dissipation kinetics of asparagine in soil measured by compound-specific analysis with metabolite tracking

    DEFF Research Database (Denmark)

    Czaban, Weronika; Rasmussen, Jim; Nicolaisen, Mogens

    2016-01-01

    formation of compounds closely related to 13C15N-asparagine metabolism over 3 h in agricultural soil. 13C15N-asparagine was rapidly dissipated with half-lives of 55 and 231 min for the low and high 13C15N-asparagine concentrations used. Universally labeled aspartic acid and interestingly also universally......Estimating the potential for direct plant acquisition of organic N, in particular amino acids, requires assessment of their turnover times in soil. It is well known from 14C studies that mineralization of amino acids occurs within hours, but mineralization to 14CO2 does not indicate the rate...... of disappearance of the intact amino acid or the possible formation of metabolites during amino acid dissipation. We here used compound-specific isotope analysis with metabolite tracking to investigate the dissipation rate of universally labeled intact 13C15N-asparagine at two concentrations and the subsequent...

  4. Direct Immunofluorescence in Oral Lichen Planus

    Science.gov (United States)

    Okuma, Nis; Thanakun, Supanee; Laothumthut, Titikarn

    2015-01-01

    Introduction Oral lichen planus (OLP) is a common immune-mediated oral mucosal disease. Diagnosis of OLP depends mainly on both clinical and histopathological features. Direct immunofluorescence (DIF) is a useful investigation method to distinguish between similar lesions and to confirm diagnosis in cases of uncharacterized features. Aim The purpose of this study was to evaluate the prevalence and pattern of DIF in a group of Thai patients with OLP. Materials and Methods Records of clinically and histologically diagnosed OLP patients attending the Oral Medicine Clinic, Faculty of Dentistry, Mahidol University, Bangkok, Thailand were consecutively reviewed for DIF results. The DIF patterns in these patients were analysed. Results There were 82 atrophic and/or erosive OLP patients with a mean age of 51.6 years. Male to female ratio was 1:5. Of these, 82.9% showed positive DIF. Buccal mucosa was superior to the gingiva and palate in terms of sensitivity for DIF. All specimens except one (98.5%) demonstrated deposition of fibrinogen at the basement membrane zone (BMZ) in a shaggy pattern. The most common DIF pattern was shaggy fibrinogen at BMZ with IgM deposition on the colloid bodies (CB) (35.3%) followed by shaggy fibrinogen along BMZ (27.9%). Conclusion The prevalence of positive DIF in Thai OLP patients was 82.9%. The most common finding was shaggy fibrinogen at BMZ. The typical pattern was shaggy fibrinogen along BMZ with or without positive IgM at CB. DIF pattern could be evaluated for the diagnosis of OLP lacking clinical and/or histopathological characteristic features. PMID:26436043

  5. Identification of proteins in painting cross-sections by immunofluorescence microscopy.

    Science.gov (United States)

    Vagnini, M; Pitzurra, L; Cartechini, L; Miliani, C; Brunetti, B G; Sgamellotti, A

    2008-09-01

    Immunofluorescence microscopy offers a highly specific analytical tool for unambiguous recognition and mapping of proteins in complex matrices. In the present work, the analytical potentials of immunofluorescence microscopy have been exploited to provide recognition of proteinaceous binders in painting cross-sections. An optimised analytical protocol is proposed for the identification of ovalbumin and of bovine serum albumin as markers of egg white and casein, respectively. The study has been carried out on laboratory model samples simulating both easel and mural paintings. The obtained results demonstrated the effectiveness of the method, suggesting the potential future use of immunofluorescence microscopy as a routine diagnostic tool in conservation science. Possible developments of the proposed methodology in order to improve the specificity of the method and its detection sensitivity are presented and discussed.

  6. In Situ and Laboratory Studies on the Fate of Specific Organic Compounds in an Anerobic Landfill Leachate Plume, 1. Experimental Conditions and Fate of Phenolic Compounds

    DEFF Research Database (Denmark)

    Nielsen, Per Henning; Albrechtsen, Hans-Jørgen; Heron, Gorm

    1995-01-01

    microcosm experiments performed and the results on the fate of 7 phenolic compounds. Part 2 of this series of papers, also published in this issue, presents the results on the fate of 8 aromatic compounds and 4 chlorinated aliphatic compounds. The redox conditions in the plume were characterized...... by in situ and laboratory experiments, both concerning redox conditions and the fate of the phenolic compounds. However, for phenol and 2,4-dichlorophenol, transformation was observed in some in situ experiments but not in the corresponding laboratory experiments. In some experiments, this could be explained......The transformation of specific organic compounds was investigated by in situ and laboratory experiments in an anaerobic landfill leachate pollution plume at four different distances from the landfill. This paper presents the experimental conditions in the in situ microcosm and laboratory batch...

  7. Potential impurities in drug substances: Compound-specific toxicology limits for 20 synthetic reagents and by-products, and a class-specific toxicology limit for alkyl bromides.

    Science.gov (United States)

    Bercu, J P; Galloway, S M; Parris, P; Teasdale, A; Masuda-Herrera, M; Dobo, K; Heard, P; Kenyon, M; Nicolette, J; Vock, E; Ku, W; Harvey, J; White, A; Glowienke, S; Martin, E A; Custer, L; Jolly, R A; Thybaud, V

    2018-02-09

    This paper provides compound-specific toxicology limits for 20 widely used synthetic reagents and common by-products that are potential impurities in drug substances. In addition, a 15 μg/day class-specific limit was developed for monofunctional alkyl bromides, aligning this with the class-specific limit previously defined for monofunctional alkyl chlorides. Both the compound- and class-specific toxicology limits assume a lifetime chronic exposure for the general population (including sensitive subpopulations) by all routes of exposure for pharmaceuticals. Inhalation-specific toxicology limits were also derived for acrolein, formaldehyde, and methyl bromide because of their localized toxicity via that route. Mode of action was an important consideration for a compound-specific toxicology limit. Acceptable intake (AI) calculations for certain mutagenic carcinogens assumed a linear dose-response for tumor induction, and permissible daily exposure (PDE) determination assumed a non-linear dose-response. Several compounds evaluated have been previously incorrectly assumed to be mutagenic, or to be mutagenic carcinogens, but the evidence reported here for such compounds indicates a lack of mutagenicity, and a non-mutagenic mode of action for tumor induction. For non-mutagens with insufficient data to develop a toxicology limit, the ICH Q3A qualification thresholds are recommended. The compound- and class-specific toxicology limits described here may be adjusted for an individual drug substance based on treatment duration, dosing schedule, severity of the disease and therapeutic indication. Copyright © 2018. Published by Elsevier Inc.

  8. Lichen specific thallus mass and secondary compounds change across a retrogressive fire-driven chronosequence.

    Directory of Open Access Journals (Sweden)

    Johan Asplund

    Full Text Available In the long-term absence of major disturbances ecosystems enter a state of retrogression, which involves declining soil fertility and consequently a reduction in decomposition rates. Recent studies have looked at how plant traits such as specific leaf mass and amounts of secondary compounds respond to declining soil fertility during retrogression, but there are no comparable studies for lichen traits despite increasing recognition of the role that lichens can play in ecosystem processes. We studied a group of 30 forested islands in northern Sweden differing greatly in fire history, and collectively representing a retrogressive chronosequence, spanning 5000 years. We used this system to explore how specific thallus mass (STM and carbon based secondary compounds (CBSCs change in three common epiphytic lichen species (Hypogymnia phsyodes, Melanohalea olivacea and Parmelia sulcata as soil fertility declines during this retrogression. We found that STMs of lichens increased sharply during retrogression, and for all species soil N to P ratio (which increased during retrogression was a strong predictor of STM. When expressed per unit area, medullary CBSCs in all species and cortical CBSCs in P. sulcata increased during retrogression. Meanwhile, when expressed per unit mass, only cortical CBSCs in H. physodes responded to retrogression, and in the opposite direction. Given that lichen functional traits are likely to be important in driving ecological processes that drive nutrient and carbon cycling in the way that plant functional traits are, the changes that they undergo during retrogression could potentially be significant for the functioning of the ecosystem.

  9. Lichen Specific Thallus Mass and Secondary Compounds Change across a Retrogressive Fire-Driven Chronosequence

    Science.gov (United States)

    Asplund, Johan; Sandling, Aron; Wardle, David A.

    2012-01-01

    In the long-term absence of major disturbances ecosystems enter a state of retrogression, which involves declining soil fertility and consequently a reduction in decomposition rates. Recent studies have looked at how plant traits such as specific leaf mass and amounts of secondary compounds respond to declining soil fertility during retrogression, but there are no comparable studies for lichen traits despite increasing recognition of the role that lichens can play in ecosystem processes. We studied a group of 30 forested islands in northern Sweden differing greatly in fire history, and collectively representing a retrogressive chronosequence, spanning 5000 years. We used this system to explore how specific thallus mass (STM) and carbon based secondary compounds (CBSCs) change in three common epiphytic lichen species (Hypogymnia phsyodes, Melanohalea olivacea and Parmelia sulcata) as soil fertility declines during this retrogression. We found that STMs of lichens increased sharply during retrogression, and for all species soil N to P ratio (which increased during retrogression) was a strong predictor of STM. When expressed per unit area, medullary CBSCs in all species and cortical CBSCs in P. sulcata increased during retrogression. Meanwhile, when expressed per unit mass, only cortical CBSCs in H. physodes responded to retrogression, and in the opposite direction. Given that lichen functional traits are likely to be important in driving ecological processes that drive nutrient and carbon cycling in the way that plant functional traits are, the changes that they undergo during retrogression could potentially be significant for the functioning of the ecosystem. PMID:23145078

  10. Increase in the specific radioactivity of tritium-labeled compounds obtained by tritium thermal activation method

    Energy Technology Data Exchange (ETDEWEB)

    Badun, G.A.; Chernysheva, M.G. [Lomonosov Moscow State Univ., Moscow (Russian Federation). Radiochemistry Div.; Ksenofontov, A.L. [Lomonosov Moscow State Univ., Moscow (Russian Federation). N. Belozersky Institute of Physico-Chemical Biology

    2012-07-01

    A method of tritium introduction into different types of organic molecules that is based on the interaction of atomic tritium with solid organic target is described. Tritium atoms are formed on the hot W-wire, which is heated by the electric current. Such an approach is called 'tritium thermal activation method'. Here we summarize the results of labeling globular proteins (lysozyme, human and bovine serum albumins); derivatives of pantothenic acid and amino acids; ionic surfactants (sodium dodecylsulfate and alkyltrimethylammonium bromides) and nonionic high-molecular weight surfactants - pluronics. For the first time it is observed that if the target-compound is fixed and its radicals are stable the specific radioactivity of the labeled product can be drastically increased (up to 400 times) when the target temperature is ca. 295 K compared with the results obtained at 77 K. The influence of labeling parameters as tritium gas pressure, exposure time and W-wire temperature was tested for each target temperature that results in the optimum labeling conditions with high specific radioactivity and chemical yield of the resulting compound. (orig.)

  11. Halocarbon emissions by selected tropical seaweeds: species-specific and compound-specific responses under changing pH

    Directory of Open Access Journals (Sweden)

    Paramjeet Kaur Mithoo-Singh

    2017-01-01

    Full Text Available Five tropical seaweeds, Kappaphycus alvarezii (Doty Doty ex P.C. Silva, Padina australis Hauck, Sargassum binderi Sonder ex J. Agardh (syn. S. aquifolium (Turner C. Agardh, Sargassum siliquosum J. Agardh and Turbinaria conoides (J. Agardh Kützing, were incubated in seawater of pH 8.0, 7.8 (ambient, 7.6, 7.4 and 7.2, to study the effects of changing seawater pH on halocarbon emissions. Eight halocarbon species known to be emitted by seaweeds were investigated: bromoform (CHBr3, dibro­momethane (CH2Br2, iodomethane (CH3I, diiodomethane (CH2I2, bromoiodomethane (CH2BrI, bromochlorometh­ane (CH2BrCl, bromodichloromethane (CHBrCl2, and dibro­mochloromethane (CHBr2Cl. These very short-lived halocarbon gases are believed to contribute to stratospheric halogen concentrations if released in the tropics. It was observed that the seaweeds emit all eight halocarbons assayed, with the exception of K. alvarezii and S. binderi for CH2I2 and CH3I respectively, which were not measurable at the achievable limit of detection. The effect of pH on halocarbon emission by the seaweeds was shown to be species-specific and compound specific. The highest percentage changes in emissions for the halocarbons of interest were observed at the lower pH levels of 7.2 and 7.4 especially in Padina australis and Sargassum spp., showing that lower seawater pH causes elevated emissions of some halocarbon compounds. In general the seaweed least affected by pH change in terms of types of halocarbon emission, was P. australis. The commercially farmed seaweed K. alvarezii was very sensitive to pH change as shown by the high increases in most of the compounds in all pH levels relative to ambient. In terms of percentage decrease in maximum quantum yield of photosynthesis (Fv∕Fm prior to and after incubation, there were no significant correlations with the various pH levels tested for all seaweeds. The correlation between percentage decrease in the maximum quantum yield of

  12. Compound-Specific Isotope Analysis of Diesel Fuels in a Forensic Investigation

    Directory of Open Access Journals (Sweden)

    Syahidah Akmal Muhammad

    2015-02-01

    Full Text Available Compound-specific isotope analysis (CSIA offers great potential as a tool to provide chemical evidence in a forensic investigation. Many attempts to trace environmental oil spills were successful where isotopic values were particularly distinct. However, difficulties arise when a large data set is analyzed and the isotopic differences between samples are subtle. In the present study, discrimination of diesel oils involved in a diesel theft case was carried out to infer the relatedness of the samples to potential source samples. This discriminatory analysis used a suite of hydrocarbon diagnostic indices, alkanes, to generate carbon and hydrogen isotopic data of the compositions of the compounds which were then processed using multivariate statistical analyses to infer the relatedness of the data set. The results from this analysis were put into context by comparing the data with the δ13C and δ2H of alkanes in commercial diesel samples obtained from various locations in the South Island of New Zealand. Based on the isotopic character of the alkanes, it is suggested that diesel fuels involved in the diesel theft case were distinguishable. This manuscript shows that CSIA when used in tandem with multivariate statistical analysis provide a defensible means to differentiate and source-apportion qualitatively similar oils at the molecular level. This approach was able to overcome confounding challenges posed by the near single-point source of origin i.e. the very subtle differences in isotopic values between the samples.

  13. Demonstration of compound-specific isotope analysis of hydrogen isotope ratios in chlorinated ethenes.

    Science.gov (United States)

    Kuder, Tomasz; Philp, Paul

    2013-02-05

    High-temperature pyrolysis conversion of organic analytes to H(2) in hydrogen isotope ratio compound-specific isotope analysis (CSIA) is unsuitable for chlorinated compounds such as trichloroethene (TCE) and cis-1,2-dichloroethene (DCE), due to competition from HCl formation. For this reason, the information potential of hydrogen isotope ratios of chlorinated ethenes remains untapped. We present a demonstration of an alternative approach where chlorinated analytes reacted with chromium metal to form H(2) and minor amounts of HCl. The values of δ(2)H were obtained at satisfactory precision (± 10 to 15 per thousand), however the raw data required daily calibration by TCE and/or DCE standards to correct for analytical bias that varies over time. The chromium reactor has been incorporated into a purge and trap-CSIA method that is suitable for CSIA of aqueous environmental samples. A sample data set was obtained for six specimens of commercial product TCE. The resulting values of δ(2)H were between -184 and +682 ‰, which significantly widened the range of manufactured TCE δ(2)H signatures identified by past work. The implications of this finding to the assessment of TCE contamination are discussed.

  14. Breast cancer chemoprevention by dietary natural phenolic compounds: specific epigenetic related molecular targets.

    Science.gov (United States)

    Pan, Min-Hsiung; Chiou, Yi-Siou; Chen, Li-Hua; Ho, Chi-Tang

    2015-01-01

    Breast cancer is a systemic malignant disease that is a major cause of cancer-related death among women worldwide. Recently, multiple lines of evidence from epidemiologic studies have suggested that epigenetic and genetic changes are involved in breast cancer development. In breast cancer patients, hormone receptor status, breast cancer stem-like cell population, and tumor microenvironment are reflective of breast cancer progression, drug resistance, and tumor recurrence. Strong relationships between a phytochemical-rich diet and a reversal of epigenetic alterations and/or modulated signaling pathways of carcinogenesis (initiation, promotion, and progression) suggest a potential approach for preventing breast cancer. Additionally, dietary consumption of natural phenolic compounds containing phytoestrogen properties exerts beneficial effects in breast cancer chemoprevention. In this review, we summarize the specific chemopreventive targets of representative phenolic compounds with an emphasis on their efficacy at interfering with epigenetic event related hormonal and nonhormonal signaling cascades that are responsible for multistage breast carcinogenesis. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  15. Source apportionment of elevated wintertime PAHs by compound-specific radiocarbon analysis

    Directory of Open Access Journals (Sweden)

    R. J. Sheesley

    2009-05-01

    Full Text Available Natural abundance radiocarbon analysis facilitates distinct source apportionment between contemporary biomass/biofuel (14C "alive" versus fossil fuel (14C "dead" combustion. Here, the first compound-specific radiocarbon analysis (CSRA of atmospheric polycyclic aromatic hydrocarbons (PAHs was demonstrated for a set of samples collected in Lycksele, Sweden a small town with frequent episodes of severe atmospheric pollution in the winter. Renewed interest in using residential wood combustion (RWC means that this type of seasonal pollution is of increasing concern in many areas. Five individual/paired PAH isolates from three pooled fortnight-long filter collections were analyzed by CSRA: phenanthrene, fluoranthene, pyrene, benzo[b+k]fluoranthene and indeno[cd]pyrene plus benzo[ghi]perylene; phenanthrene was the only compound also analyzed in the gas phase. The measured Δ14C for PAHs spanned from −138.3‰ to 58.0‰. A simple isotopic mass balance model was applied to estimate the fraction biomass (fbiomass contribution, which was constrained to 71–87% for the individual PAHs. Indeno[cd]pyrene plus benzo[ghi]perylene had an fbiomass of 71%, while fluoranthene and phenanthrene (gas phase had the highest biomass contribution at 87%. The total organic carbon (TOC, defined as carbon remaining after removal of inorganic carbon fbiomass was estimated to be 77%, which falls within the range for PAHs. This CSRA data of atmospheric PAHs established that RWC is the dominating source of atmospheric PAHs to this region of the boreal zone with some variations among RWC contributions to specific PAHs.

  16. A Novel Benzodiazepine Compound Inhibits Yellow Fever Virus Infection by Specifically Targeting NS4B Protein.

    Science.gov (United States)

    Guo, Fang; Wu, Shuo; Julander, Justin; Ma, Julia; Zhang, Xuexiang; Kulp, John; Cuconati, Andrea; Block, Timothy M; Du, Yanming; Guo, Ju-Tao; Chang, Jinhong

    2016-09-21

    -risk regions. It has been estimated that up to 1.7 million YFV infections occur in Africa each year, resulting in 29,000 to 60,000 death. Thus far, there is no specific antiviral treatment for yellow fever. To cope with this medical challenge, we identified a benzodiazepine compound that selectively inhibits YFV by targeting the viral NS4B protein. To our knowledge, this is the first report demonstrating in vivo safety and antiviral efficacy of an YFV NS4B inhibitor in an animal model. We have thus reached a critical milestone toward the development of specific antiviral therapeutics for clinical management of yellow fever. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  17. Bioaccumulation of photoprotective compounds in copepods: environmental triggers and sources of intra-specific variability

    Science.gov (United States)

    Zagarese, H. E.; García, P.; Diéguez, M. D.; Ferraro, M. A.

    2012-12-01

    Ultraviolet radiation (UVR) and temperature are two globally important abiotic factors affecting freshwater ecosystems. Planktonic organisms have developed a battery of counteracting mechanisms to minimize the risk of being damaged by UVR, which respond to three basic principles: avoid, protect, repair. Copepods are among the most successful zooplankton groups. They are highly adaptable animals, capable of displaying flexible behaviors, physiologies, and life strategies. In particular, they are well equipped to cope with harmful UVR. Their arsenal includes vertical migration, accumulation of photoprotective compounds, and photorepair. The preference for a particular strategy is affected by a plethora of environmental (extrinsic) parameters, such as the existence of a depth refuge, the risk of visual predation, and temperature. Temperature modifies the environment (e.g. the lake thermal structure), and animal metabolism (e.g., swimming speed, bioaccumulation of photoprotective compounds). In addition, the relative weight of UVR-coping strategies is also influenced by the organism (intrinsic) characteristics (e.g., inter- and intra-specific variability). The UV absorbing compounds, mycosporine-like amino acids (MAAs), are widely distributed among freshwater copepods. Animals are unable to synthesize MAAs, and therefore depend on external sources for accumulating these compounds. Although copepods may acquire MAAs from their food, for the few centropagic species investigated so far, the main source of MAAs are microbial (most likely prokaryotic) organisms living in close association with the copepods. Boeckella gracilipes is a common centropagic copepod in Patagonian lakes. We suspected that its occurrence in different types of lakes, hydrologically unconnected, but within close geographical proximity, could have resulted in different microbial-copepod associations (i.e., different MAAs sources) that could translate into intra-specific differences in the accumulation

  18. Halenaquinone, a chemical compound that specifically inhibits the secondary DNA binding of RAD51.

    Science.gov (United States)

    Takaku, Motoki; Kainuma, Takashi; Ishida-Takaku, Takako; Ishigami, Shintaro; Suzuki, Hidekazu; Tashiro, Satoshi; van Soest, Rob W M; Nakao, Yoichi; Kurumizaka, Hitoshi

    2011-04-01

    Mutations and single-nucleotide polymorphisms affecting RAD51 gene function have been identified in several tumors, suggesting that the inappropriate expression of RAD51 activity may cause tumorigenesis. RAD51 is an essential enzyme for the homologous recombinational repair (HRR) of DNA double-strand breaks. In the HRR pathway, RAD51 catalyzes the homologous pairing between single-stranded DNA (ssDNA) and double-stranded DNA (dsDNA), which is the central step of the HRR pathway. To identify a chemical compound that regulates the homologous-pairing activity of RAD51, in the present study, we screened crude extract fractions from marine sponges by the RAD51-mediated homologous-pairing assay. Halenaquinone was identified as an inhibitor of the RAD51 homologous-pairing activity. A surface plasmon resonance analysis indicated that halenaquinone directly bound to RAD51. Intriguingly, halenaquinone specifically inhibited dsDNA binding by RAD51 alone or the RAD51-ssDNA complex, but only weakly affected the RAD51-ssDNA binding. In vivo, halenaquinone significantly inhibited the retention of RAD51 at double-strand break sites. Therefore, halenaquinone is a novel type of RAD51 inhibitor that specifically inhibits the RAD51-dsDNA binding. © 2011 The Authors. Journal compilation © 2011 by the Molecular Biology Society of Japan/Blackwell Publishing Ltd.

  19. Flow cytometric immunofluorescence of rat anterior pituitary cells

    Science.gov (United States)

    Hatfield, J. Michael; Hymer, W. C.

    1985-01-01

    A flow cytometric immunofluorescence technique was developed for the quantification of growth hormone, prolactin, and luteinizing hormone producing cells. The procedure is based on indirect-immunofluorescence of intracellular hormone using an EPICS V cell sorter and can objectively count 50,000 cells in about 3 minutes. It can be used to study the dynamics of pituitary cell populations under various physiological and pharmacological conditions.

  20. Changes in floral bouquets from compound-specific responses to increasing temperatures.

    Science.gov (United States)

    Farré-Armengol, Gerard; Filella, Iolanda; Llusià, Joan; Niinemets, Ulo; Peñuelas, Josep

    2014-12-01

    We addressed the potential effects of changes in ambient temperature on the profiles of volatile emissions from flowers and tested whether warming could induce significant quantitative and qualitative changes in floral emissions, which would potentially interfere with plant-pollinator chemical communication. We measured the temperature responses of floral emissions of various common species of Mediterranean plants using dynamic headspace sampling and used GC-MS to identify and quantify the emitted terpenes. Floral emissions increased with temperature to an optimum and thereafter decreased. The responses to temperature modeled here predicted increases in the rates of floral terpene emission of 0.03-1.4-fold, depending on the species, in response to an increase of 1 °C in the mean global ambient temperature. Under the warmest projections that predict a maximum increase of 5 °C in the mean temperature of Mediterranean climates in the Northern Hemisphere by the end of the century, our models predicted increases in the rates of floral terpene emissions of 0.34-9.1-fold, depending on the species. The species with the lowest emission rates had the highest relative increases in floral terpene emissions with temperature increases of 1-5 °C. The response of floral emissions to temperature differed among species and among different compounds within the species. Warming not only increased the rates of total emissions, but also changed the ratios among compounds that constituted the floral scents, i.e. increased the signal for pollinators, but also importantly altered the signal fidelity and probability of identification by pollinators, especially for specialists with a strong reliance on species-specific floral blends. © 2014 John Wiley & Sons Ltd.

  1. Compound-Specific Isotope Analysis of Nitroaromatic Contaminant Transformations by Nitroarene Dioxygenases

    Science.gov (United States)

    Pati, Sarah G.; Kohler, Hans-Peter E.; Hofstetter, Thomas B.

    2014-05-01

    Dioxygenation is an important biochemical reaction that often initiates the mineralization of recalcitrant organic contaminants such as nitroaromatic explosives, chlorinated benzenes, and polycyclic aromatic hydrocarbons. However, to assess the extent of dioxygenation in contaminated environments is difficult because of competing transformation processes and further reactions of the dioxygenation products. Compound-specific isotope analysis (CSIA) offers a new approach to reliably quantify biodegradation initiated by dioxygenation based on changes in stable isotope ratios of the pollutant. For CSIA it is essential to know the kinetic isotope effects (KIEs) pertinent to the dioxygenation mechanism of organic contaminants. Unfortunately, the range of KIEs of such reactions is poorly constrained although many dioxygenase enzymes with a broad substrate specificity have been reported. Dioxygenase enzymes usually exhibit complex reaction kinetics involving multiple substrates and substrate-specific binding modes which makes the determination of KIEs challenging. The goal of this study was to explore the magnitude and variability of 13C-, 2H-, and 15N-KIEs for the dioxygenation of one contaminant class, that is nitroaromatic contaminants (NACs). To this end, we investigated the C, H, and N isotope fractionation during the dioxygenation of nitrobenzene (NB), 2-nitrotoluene (2-NT), and 3-nitrotoluene (3-NT) by pure cultures, E. coli clones, cell extracts, and purified enzymes. From isotope fractionations measured in the substrates and reaction products, we determined dioxygenation KIEs for different combinations of the three substrates with nitrobenzene dioxygenase (NBDO) and 2-nitrotoluene dioxygenase (2NTDO). The 13C-, 2H-, and 15N-KIEs for the dioxygenation of NB by NBDO were consistent for all experimental systems considered (i.e., Comamonas sp. Strain JS765, E. coli clones, cell extracts of E. coli clones, and purified NBDO). This observation suggests that the isotope

  2. Intracellular distribution of Tankyrases as detected by multicolor immunofluorescence techniques

    Directory of Open Access Journals (Sweden)

    M.G. Bottone

    2012-01-01

    Full Text Available Poly(ADP-ribose polymerases are a family of enzymes that catalyze the conversion of NAD+ into ADP-ribose. Among them, Tankyrases have been found to bind to centrosome, mitotic spindle and microsome proteins, in the cytoplasm, and to telomeres in the nucleus, where they play a relevant role in telomere metabolism. However, their precise intracellular localization during interphase has not been so far fully elucidated. We investigated this aspect in situ by double immunofluorescence experiments using antibodies recognizing Tankyrases 1-2 or other proteins residing in specific organelles (Golgi apparatus, mitochondria, lysosomes, endoplasmic reticulum. We used HeLa cells as a model system in vitro, before and after treatment with either actinomycin D or etoposide, to also investigate the possible relocation of Tankyrases during apoptosis. We observed that Tankyrases are distributed both in the nucleus and in the cytoplasm; in this latter compartment, they were found to colocate with the Golgi apparatus but never with the mitochondria; a pool of Tankyrases also colocates with the endoplasmic reticulum and lysosomes. Interestingly, in cells with clear signs of apoptosis, Tankyrases were detectable in the cytoplasmic blebs: this suggests that they are not massively cleaved during apoptosis and persist in the largely heterogeneous apoptotic remnants which are known to contain components of cytoplasmic and nuclear origin.

  3. 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.

  4. Detection of Tilletia controversa using immunofluorescent monoclonal antibodies.

    Science.gov (United States)

    Gao, L; Feng, C; Li, B; Liu, T; Liu, B; Chen, W

    2015-02-01

    Tilletia controversa is an internationally quarantined pathogenic fungus that causes dwarf bunt of wheat and is similar to Tilletia caries in both teliospore morphology and genetic structure. This study developed a rapid and sensitive immunofluorescence method for differentiating the teliospores of T. controversa from T. caries. The method utilizes monoclonal antibody D-1 against teliospores of T. controversa as well as a PE-Cy3-conjugated goat anti-mouse antibody (overlapping light excitation of 495 and 555 nm). The orange cycle fluorescent signal was stronger against T. controversa teliospores in the outer spore wall and net ridge, whereas only the green signal was observed for the protoplasm of T. caries teliospores. The detection limit of this method was 2.0 μg ml(-1) of the D-1 monoclonal antibody. This study describes the production and diagnostic application of a novel mouse monoclonal antibody specific to T. controversa teliospores. This method could be used for the on-site identification of T. controversa teliospores in the near future and will help in selecting fungicides to control dwarf bunt of wheat as further technical developments are achieved. © 2014 The Society for Applied Microbiology.

  5. Compound-specific hydrogen isotope analysis of heteroatom-bearing compounds via gas chromatography-chromium-based high-temperature conversion (Cr/HTC)-isotope ratio mass spectrometry.

    Science.gov (United States)

    Renpenning, Julian; Kümmel, Steffen; Hitzfeld, Kristina L; Schimmelmann, Arndt; Gehre, Matthias

    2015-09-15

    The traditional high-temperature conversion (HTC) approach toward compound-specific stable isotope analysis (CSIA) of hydrogen for heteroatom-bearing (i.e., N, Cl, S) compounds has been afflicted by fractionation bias due to formation of byproducts HCN, HCl, and H2S. This study presents a chromium-based high-temperature conversion (Cr/HTC) approach for organic compounds containing nitrogen, chlorine, and sulfur. Following peak separation along a gas chromatographic (GC) column, the use of thermally stable ceramic Cr/HTC reactors at 1100-1500 °C and chemical sequestration of N, Cl, and S by chromium result in quantitative conversion of compound-specific organic hydrogen to H2 analyte gas. The overall hydrogen isotope analysis via GC-Cr/HTC-isotope ratio mass spectrometry (IRMS) achieved a precision of better than ± 5 mUr along the VSMOW-SLAP scale. The accuracy of GC-Cr/HTC-IRMS was validated with organic reference materials (RM) in comparison with online EA-Cr/HTC-IRMS and offline dual-inlet IRMS. The utility and reliability of the GC-Cr/HTC-IRMS system were documented during the routine measurement of more than 500 heteroatom-bearing organic samples spanning a δ(2)H range of -181 mUr to 629 mUr.

  6. Compound-specific chlorine isotope analysis of polychlorinated biphenyls isolated from Aroclor and Clophen technical mixtures.

    Science.gov (United States)

    Mandalakis, Manolis; Holmstrand, Henry; Andersson, Per; Gustafsson, Orjan

    2008-03-01

    Compound-specific chlorine isotope analysis (CSIA-Cl) is promising as a novel and powerful method for monitoring in situ degradation of organochlorines in the environment and for source fingerprinting purposes. In order to apply CSIA-Cl in field studies of polychlorinated biphenyls (PCBs), the chlorine isotopic composition (delta(37)Cl) of individual PCB congeners in source materials must be known. In the present study, we determined delta(37)Cl of 18 congeners isolated from three widely produced technical mixtures. All congeners provided delta(37)Cl ranging between -1.9 per thousand and -3.5 per thousand. Although the comparable products Aroclor 1242 (-2.0 per thousand to -2.5 per thousand) and Clophen A30 (-1.9 per thousand to -3.0 per thousand) were synthesized by different industries, they provided similar delta(37)Cl for the same type of congeners. On the contrary, the more chlorinated congeners present in Aroclor 1254 (-2.1 to -3.5 per thousand) were more (37)Cl depleted compared to Aroclor 1242 manufactured by the same company. Overall, delta(37)Cl of PCB congeners decreased by -0.26 per thousand for each additional chlorine atom.

  7. Biotransformation of benzotriazoles: insights from transformation product identification and compound-specific isotope analysis.

    Science.gov (United States)

    Huntscha, Sebastian; Hofstetter, Thomas B; Schymanski, Emma L; Spahr, Stephanie; Hollender, Juliane

    2014-04-15

    Benzotriazoles are widely used domestic and industrial corrosion inhibitors and have become omnipresent organic micropollutants in the aquatic environment. Here, the range of aerobic biological degradation mechanisms of benzotriazoles in activated sludge was investigated. Degradation pathways were elucidated by identifying transient and persistent transformation products in batch experiments using liquid chromatography-high-resolution tandem mass spectrometry (LC-HR-MS/MS). In addition, initial reactions were studied using compound-specific isotope analysis (CSIA). Biodegradation half-lives of 1.0 days for 1H-benzotriazole, 8.5 days for 4-methyl-1H-benzotriazole, and 0.9 days for 5-methyl-1H-benzotriazole with activated sludge confirmed their known partial persistence in conventional wastewater treatment. Major transformation products were identified as 4- and 5-hydroxy-1H-benzotriazole for the degradation of 1H-benzotriazole, and 1H-benzotriazole-5-carboxylic acid for the degradation of 5-methyl-1H-benzotriazole. These transformation products were found in wastewater effluents, showing their environmental relevance. Many other candidate transformation products, tentatively identified by interpretation of HR-MS/MS spectra, showed the broad range of possible reaction pathways including oxidation, alkylation, hydroxylation and indicate the significance of cometabolic processes for micropollutant degradation in biological wastewater treatment in general. The combination of evidence from product analysis with the significant carbon and nitrogen isotope fractionation suggests that aromatic monohydroxylation is the predominant step during the biotransformation of 1H-benzotriazole.

  8. Site-specific acid-base properties of pholcodine and related compounds.

    Science.gov (United States)

    Kovács, Z; Hosztafi, S; Noszál, B

    2006-11-01

    The acid-base properties of pholcodine, a cough-depressant agent, and related compounds including metabolites were studied by 1H NMR-pH titrations, and are characterised in terms of macroscopic and microscopic protonation constants. New N-methylated derivatives were also synthesized in order to quantitate site- and nucleus-specific protonation shifts and to unravel microscopic acid-base equilibria. The piperidine nitrogen was found to be 38 and 400 times more basic than its morpholine counterpart in pholcodine and norpholcodine, respectively. The protonation data show that the molecule of pholcodine bears an average of positive charge of 1.07 at physiological pH, preventing it from entering the central nervous system, a plausible reason for its lack of analgesic or addictive properties. The protonation constants of pholcodine and its derivatives are interpreted by comparing with related molecules of pharmaceutical interest. The pH-dependent relative concentrations of the variously protonated forms of pholcodine and morphine are depicted in distribution diagrams.

  9. Microbial degradation of alpha-cypermethrin in soil by compound-specific stable isotope analysis.

    Science.gov (United States)

    Xu, Zemin; Shen, Xiaoli; Zhang, Xi-Chang; Liu, Weiping; Yang, Fangxing

    2015-09-15

    To assess microbial degradation of alpha-cypermethrin in soil, attenuation of alpha-cypermethrin was investigated by compound-specific stable isotope analysis. The variations of the residual concentrations and stable carbon isotope ratios of alpha-cypermethrin were detected in unsterilized and sterilized soils spiked with alpha-cypermethrin. After an 80 days' incubation, the concentrations of alpha-cypermethrin decreased to 0.47 and 3.41 mg/kg in the unsterilized soils spiked with 2 and 10 mg/kg, while those decreased to 1.43 and 6.61 mg/kg in the sterilized soils. Meanwhile, the carbon isotope ratios shifted to -29.14 ± 0.22‰ and -29.86 ± 0.33‰ in the unsterilized soils spiked with 2 and 10 mg/kg, respectively. The results revealed that microbial degradation contributed to the attenuation of alpha-cypermethrin and induced the carbon isotope fractionation. In order to quantitatively assess microbial degradation, a relationship between carbon isotope ratios and residual concentrations of alpha-cypermethrin was established according to Rayleigh equation. An enrichment factor, ϵ = -1.87‰ was obtained, which can be employed to assess microbial degradation of alpha-cypermethrin. The significant carbon isotope fractionation during microbial degradation suggests that CSIA is a proper approach to qualitatively detect and quantitatively assess the biodegradation during attenuation process of alpha-cypermethrin in the field. Copyright © 2015 Elsevier B.V. All rights reserved.

  10. Compound-Specific Stable Isotope Fractionation of Pesticides and Pharmaceuticals in a Mesoscale Aquifer Model.

    Science.gov (United States)

    Schürner, Heide K V; Maier, Michael P; Eckert, Dominik; Brejcha, Ramona; Neumann, Claudia-Constanze; Stumpp, Christine; Cirpka, Olaf A; Elsner, Martin

    2016-06-07

    Compound-specific isotope analysis (CSIA) receives increasing interest for its ability to detect natural degradation of pesticides and pharmaceuticals. Despite recent laboratory studies, CSIA investigations of such micropollutants in the environment are still rare. To explore the certainty of information obtainable by CSIA in a near-environmental setting, a pulse of the pesticide bentazone, the pesticide metabolite 2,6-dichlorobenzamide (BAM), and the pharmaceuticals diclofenac and ibuprofen was released into a mesoscale aquifer with quasi-two-dimensional flow. Concentration breakthrough curves (BTC) of BAM and ibuprofen demonstrated neither degradation nor sorption. Bentazone was transformed but did not sorb significantly, whereas diclofenac showed both degradation and sorption. Carbon and nitrogen CSIA could be accomplished in similar concentrations as for "traditional" priority pollutants (low μg/L range), however, at the cost of uncertainties (0.4-0.5‰ (carbon), 1‰ (nitrogen)). Nonetheless, invariant carbon and nitrogen isotope values confirmed that BAM was neither degraded nor sorbed, while significant enrichment of (13)C and in particular (15)N corroborated transformation of diclofenac and bentazone. Retardation of diclofenac was reflected in additional (15)N sorption isotope effects, whereas isotope fractionation of transverse dispersion could not be identified. These results provide a benchmark on the performance of CSIA to monitor the reactivity of micropollutants in aquifers and may guide future efforts to accomplish CSIA at even lower concentrations (ng/L range).

  11. Effects of decalcification on bulk and compound-specific nitrogen and carbon isotope analyses of dentin.

    Science.gov (United States)

    Brault, Emily K; Koch, Paul L; Gier, Elizabeth; Ruiz-Cooley, R I; Zupcic, Jessica; Gilbert, Kwasi N; McCarthy, Matthew D

    2014-12-30

    For bulk carbon and nitrogen isotope analysis of dentin, samples are typically decalcified. Since the non-protein carbon in dentin is low, whole sample analysis may produce reliable data. Compound-specific isotope analysis (CSIA) of bone and tooth dentin protein is a powerful tool for reconstructing the flow of carbon and nitrogen in modern and past food webs. Decalcification has also been used to prepare bone and dentin samples for CSIA, but the effects of this process on bulk dentin, amino acid composition, and their specific isotope values are not known. The bulk isotope values of raw and decalcified dentin from a sperm whale tooth were measured to determine the effects of decalcification and the accuracy of untreated dentin results. CSIA was also performed on decalcified and raw dentin to examine differences in the amino acid isotope values and molar composition between these two approaches. Analysis of raw dentin yields precise and accurate bulk isotope measurements for this animal. The isotopic values of decalcified samples and raw dentin for individual amino acids were similar, but the average of the isotope value offsets between the two sample types was significant. The presence of inorganic material complicated raw sample processing for individual amino acid isotope values, and may have contributed to the isotopic differences between decalcified and raw samples. Decalcification is not needed to measure bulk isotope values in dentin from this modern odontocete, probably because the lipid and carbonate concentrations are low and the carbon isotope values of dentin protein and carbonate are similar. This method should not be applied in some cases (e.g., with fossil dentin and modern bone). Decalcification should still be used prior to CSIA since significant matrix issues occur with raw dentin processing and decalcification does not alter the amino acid molar composition or isotopic values of dentin. Copyright © 2014 John Wiley & Sons, Ltd.

  12. Immunofluorescence in cytogenetic analysis: method and applications

    Directory of Open Access Journals (Sweden)

    Jeppesen Peter

    2000-01-01

    Full Text Available Control of the genetic information encoded by DNA in mammalian chromosomes is mediated by proteins, some of which are only transiently attached, although others are intrinsically associated with nucleic acid in the complex mixture known as chromatin. Chromatin-associated proteins range from the ubiquitous and abundant histones down to the most specific and rare of transcription factors. Although many chromatin proteins are probably excluded from highly condensed mitotic chromosomes, a number are retained throughout the cell cycle and can be detected on chromosomes in metaphase spreads. Comparing the distribution of a chromosomal protein with known cytogenetic markers on metaphase chromosomes can provide an important and potentially highly informative first source of data on the function of the protein under consideration. The aim of the present study is to summarize some of the principles involved in obtaining suitable chromosome preparations for subsequent immunolocalization of protein antigens. Some applications of the method will be included to illustrate how this approach has increased our understanding of chromosome structure and genetic regulation.

  13. Compound Specific Stable Chlorine Isotopic Analysis of Volatile Aliphatic Compounds Using Gas Chromatography Hyphenated with Multiple Collector Inductively Coupled Plasma Mass Spectrometry.

    Science.gov (United States)

    Horst, Axel; Renpenning, Julian; Richnow, Hans-Hermann; Gehre, Matthias

    2017-09-05

    Stable chlorine isotope analysis is increasingly used to characterize sources, transformation pathways, and sinks of organic aliphatic compounds, many of them being priority pollutants in groundwater and the atmosphere. A wider use of chlorine isotopes in environmental studies is still inhibited by limitations of the different analytical techniques such as high sample needs, offline preparation, confinement to few compounds and mediocre precision, respectively. Here we present a method for the δ37Cl determination in volatile aliphatic compounds using gas chromatography coupled with multiple-collector inductively coupled plasma mass spectrometry (GC-MC-ICPMS), which overcomes these limitations. The method was evaluated by using a suite of five previously offline characterized in-house standards and eight chlorinated methanes, ethanes, and ethenes. Other than in previous approaches using ICP methods for chlorine isotopes, isobaric interference of the 36ArH dimer with 37Cl was minimized by employing dry plasma conditions. Samples containing 2-3 nmol Cl injected on-column were sufficient to achieve a precision (σ) of 0.1 mUr (1 milliurey = 0.001 = 1‰) or better. Long-term reproducibility and accuracy was always better than 0.3 mUr if organics were analyzed in compound mixtures. Standardization is carried out by using a two-point calibration approach. Drift, even though very small in this study, is corrected by referencing versus an internal standard. The presented method offers a direct, universal, and compound-specific procedure to measure the δ37Cl of a wide array of organic compounds overcoming limitations of previous techniques with the benefits of high sensitivity and accuracy comparable to the best existing approaches.

  14. Pore-scale analysis on the effects of compound-specific dilution on transient transport and solute breakthrough

    DEFF Research Database (Denmark)

    Rolle, Massimo; Kitanidis, Peter

    breakthrough curves. Such dilution breakthrough curves allow capturing the compound-specific mixing of the different solutes and provide useful insights on the interplay between advective and diffusive processes, mass transfer limitations, and incomplete mixing in the heterogeneous pore-scale domains......Compound-specific diffusivities significantly impact solute transport and mixing at different scales. Although diffusive processes occur at the small pore scale, their effects propagate and remain important at larger macroscopic scales [1]. In this pore-scale modeling study in saturated porous...... media we show that compound-specific effects are important not only at steady-state and for the lateral displacement of solutes with different diffusivities but also for transient transport and solute breakthrough [2]. We performed flow and transport simulations in two-dimensional pore-scale domains...

  15. Immunofluorescence findings in granuloma faciale: report of two cases.

    Science.gov (United States)

    Zirwas, Matthew J; Abell, Edward; Ruben, Alan; Silverman, Alan R; Wolff, Jeffrey; Deng, Jau-Shyong

    2003-05-01

    Immunofluorescence findings in granuloma faciale have been infrequently described. Reported findings include granular IgA, IgG, IgM, and C3 deposits in the dermoepidermal junction, in blood vessel walls, and on connective tissue fibers; IgG in the basement membrane zone, and IgG around blood vessels. We report two cases of granuloma faciale in which biopsy specimens were sent for routine pathology as well as immunofluorescence. Additional work-up included serologic testing for markers of lupus erythematosus (LE). Immunofluorescence in both cases revealed granular IgA, IgG, IgM, C3, and C5-9 deposits along the dermoepidermal junction only. Blood vessel walls and connective tissue fibers were spared. Serologic studies for markers of LE were negative. Immunofluorescence findings were essentially identical to those which would be expected in cutaneous LE; however, the combination of negative serologic studies for LE, histopathologic findings typical of granuloma faciale, and clinical findings most consistent with granuloma faciale allowed the diagnosis of granuloma faciale to be rendered with certainty. Interpretation of results of direct immunofluorescence of skin should be correlated with clinical presentation, histopathological findings and other laboratory results in order to render final diagnosis of a given patient.

  16. Assessing the functional diversity of herbivorous reef fishes using a compound-specific stable isotope approach

    KAUST Repository

    Tietbohl, Matthew

    2016-12-01

    Herbivorous coral reef fishes play an important role in helping to structure their environment directly by consuming algae and indirectly by promoting coral health and growth. These fishes are generally separated into three broad groups: browsers, grazers, and excavators/scrapers, with these groupings often thought to have a fixed general function and all fishes within a group thought to have similar ecological roles. This categorization assumes a high level of functional redundancy within herbivorous fishes. However, recent evidence questions the use of this broad classification scheme, and posits that there may actually be more resource partitioning within these functional groupings. Here, I use a compound-specific stable isotope approach (CSIA) to show there appears to be a greater diversity of functional roles than previously assumed within broad functional groups. The δ13C signatures from essential amino acids of reef end-members (coral, macroalgae, detritus, and phytoplankton) and fish muscle were analyzed to investigate differences in resource use between fishes. Most end-members displayed clear isotopic differences, and most fishes within functional groups were dissimilar in their isotopic signature, implying differences in the resources they target. No grazers closely resembled each other isotopically, implying a much lower level of functional redundancy within this group; scraping parrotfish were also distinct from excavating parrotfish and to a lesser degree distinct between scrapers. This study highlights the potential of CSIA to help distinguish fine-scale ecological differences within other groups of reef organisms as well. These results question the utility of lumping nominally herbivorous fishes into broad groups with assumed similar roles. Given the apparent functional differences between nominally herbivorous reef fishes, it is important for managers to incorporate the diversity of functional roles these fish play.

  17. Fluorophilia: Fluorophore-containing compounds adhere non-specifically to injured neurons

    Science.gov (United States)

    Hawkins, Bridget E.; Frederickson, Christopher J.; DeWitt, Douglas S.; Prough, Donald S.

    2012-01-01

    Ionic (free) zinc (Zn2+) is implicated in apoptotic neuronal degeneration and death. In our attempt to examine the effects of Zn2+ in neurodegeneration following brain injury, we serendipitously discovered that injured neurons bind fluorescein moieties, either alone or as part of an indicator dye, in histologic sections. This phenomenon, that we have termed “fluorophilia”, is analogous to the ability of degenerating neuronal somata and axons to bind silver ions (argyrophilia — the basis of silver degeneration stains). To provide evidence that fluorophilia occurs in sections of brain tissue, we used a wide variety of indicators such as Fluoro-Jade (FJ), a slightly modified fluorescein sold as a marker for degenerating neurons; Newport Green, a fluorescein-containing Zn2+ probe; Rhod-5N, a rhodamine-containing Ca2+ probe; and plain fluorescein. All yielded remarkably similar staining of degenerating neurons in the traumatic brain-injured tissue with the absence of staining in our sham-injured brains. Staining of presumptive injured neurons by these agents was not modified when Zn2+ in the brain section was removed by prior chelation with EDTA or TPEN, whereas staining by a non-fluorescein containing Zn2+ probe, N-(6-methoxy-8-quinolyl)-p-toluenesulfonamide (TSQ), was suppressed by prior chelation. Thus, certain fluorophore-containing compounds nonspecifically stain degenerating neuronal tissue in histologic sections and may not reflect the presence of Zn2+. This may be of concern to researchers using indicator dyes to detect metals in brain tissue sections. Further experiments may be advised to clarify whether Zn2+-binding dyes bind more specifically in intact neurons in culture or organotypic slices. PMID:22137653

  18. Tracing carbon flow through coral reef food webs using a compound-specific stable isotope approach

    KAUST Repository

    McMahon, Kelton

    2015-11-21

    Coral reefs support spectacularly productive and diverse communities in tropical and sub-tropical waters throughout the world’s oceans. Debate continues, however, on the degree to which reef biomass is supported by new water column production, benthic primary production, and recycled detrital carbon (C). We coupled compound-specific stable C isotope ratio (δ13C) analyses with Bayesian mixing models to quantify C flow from primary producers to coral reef fishes across multiple feeding guilds and trophic positions in the Red Sea. Analyses of reef fishes with putative diets composed primarily of zooplankton (Amblyglyphidodon indicus), benthic macroalgae (Stegastes nigricans), reef-associated detritus (Ctenochaetus striatus), and coral tissue (Chaetodon trifascialis) confirmed that δ13C values of essential amino acids from all baseline C sources were both isotopically diagnostic and accurately recorded in consumer tissues. While all four source end-members contributed to the production of coral reef fishes in our study, a single-source end-member often dominated dietary C assimilation of a given species, even for highly mobile, generalist top predators. Microbially reworked detritus was an important secondary C source for most species. Seascape configuration played an important role in structuring resource utilization patterns. For instance, Lutjanus ehrenbergii showed a significant shift from a benthic macroalgal food web on shelf reefs (71 ± 13 % of dietary C) to a phytoplankton-based food web (72 ± 11 %) on oceanic reefs. Our work provides insights into the roles that diverse C sources play in the structure and function of coral reef ecosystems and illustrates a powerful fingerprinting method to develop and test nutritional frameworks for understanding resource utilization.

  19. Compound-specific stable isotopes of organic compounds from lake sediments track recent environmental changes in an alpine ecosystem, Rocky Mountain National Park, Colorado

    Science.gov (United States)

    Enders, S.K.; Pagani, M.; Pantoja, S.; Baron, J.S.; Wolfe, A.P.; Pedentchouk, N.; Nunez, L.

    2008-01-01

    Compound-specific nitrogen, carbon, and hydrogen isotope records from sediments of Sky Pond, an alpine lake in Rocky Mountain National Park (Colorado, United States of America), were used to evaluate factors contributing to changes in diatom assemblages and bulk organic nitrogen isotope records identified in lake sediments across Colorado, Wyoming, and southern Montana. Nitrogen isotopic records of purified algal chlorins indicate a substantial shift in nitrogen cycling in the region over the past ???60 yr. Temporal changes in the growth characteristics of algae, captured in carbon isotope records in and around Sky Pond, as well as a -60??? excursion in the hydrogen isotope composition of algal-derived palmitic acid, are coincident with changes in nitrogen cycling. The confluence of these trends is attributed to an increase in biologically available nitrogenous compounds caused by an expansion of anthropogenic influences and temporal changes in catchment hydrology and nutrient delivery associated with meltwater dynamics. ?? 2008, by the American Society of Limnology and Oceanography, Inc.

  20. Culture confirmation of Escherichia coli serotype O157:H7 by direct immunofluorescence.

    OpenAIRE

    Tison, D L

    1990-01-01

    An evaluation of a fluorescein-labeled, polyclonal, affinity-purified goat antibody to Escherichia coli serotype O157:H7 (Kirkegaard & Perry Laboratories Inc., Gaithersburg, Md.) was conducted to determine the efficacy of this research reagent for the rapid direct immunofluorescence identification of E. coli O157:H7 isolated from fecal specimens cultured on sorbitol-MacConkey (SMAC) agar. The E. coli O157:H7 fluorescent-antibody conjugate proved to be 100% sensitive and specific for the rapid...

  1. Structure elucidation and DNA binding specificity of natural compounds from Cassia siamea leaves: A biophysical approach.

    Science.gov (United States)

    Parveen, Mehtab; Ahmad, Faheem; Malla, Ali Mohammed; Khan, Mohd Sohrab; Rehman, Sayeed Ur; Tabish, Mohammad; Silva, Manuela Ramos; Silva, P S Pereira

    2016-06-01

    A novel isoflavone, 5,6,7-trimethoxy-3-(3',4',5'-trimethoxyphenyl)-4H-chromen-4-one (1) along with a known pyranocoumarin, Seselin (2) have been isolated from the ethanolic extract of the leaves of Cassia siamea (Family: Fabaceae). Compound 1 has been reported for the first time from any natural source and has not been synthesized so far. Their structures were elucidated on the basis of chemical and physical evidences viz. elemental analysis, UV, FT-IR, (1)H-NMR, (13)C-NMR and mass spectral analysis. Structure of compound (1) was further authenticated by single-crystal X-ray analysis and density functional theory (DFT) calculations. A multi-technique approach employing UV-Visible spectroscopy, fluorescence, KI quenching studies, competitive displacement assay, circular dichroism and viscosity studies have been utilized to probe the extent of interaction and possible binding modes of isolated compounds (1-2) with calf thymus DNA (CT-DNA). Both the compounds were found to interact with DNA via non-intercalative binding mode with moderate proficiencies. Groove binding was the major interaction mode in the case of compound 2 while compound 1 probably interacts with DNA through electrostatic interactions. These studies provide deeper insight in understanding of DNA-drug (natural products) interaction which could be helpful to improve their bioavailability for therapeutic purposes. Copyright © 2016 Elsevier B.V. All rights reserved.

  2. [Markers for Immunofluorescence Analysis Based on Europium Complex with Fluorinated β-Diketones Carbazole Series].

    Science.gov (United States)

    Kostryukova, T S; Ivanovskaya, N P; Zatonsky, G V; Osin, N S; Vasilev, N V

    2015-01-01

    In order to develop new markers for immunofluorescence time-resolved luminescence analysis tetraketodiester carbazole series was obtained. Compound contains methoxycarbonyl reactive groups, separated from the 1,3-dicarbonyl chelating fragment by difluoromethylene spacers (CF2)4. Complexes of this compound with the ions Eu3+ are stable in aqueous solution and possess a long-living intensive luminescence with the main peaks: emission--in the area of 615 nm and excitation--in the 380-390 nm, that distinguishes them from most used analogues. Conjugate of streptavidin to compound was obtained and its fluorescent spectral characteristics allow to use it as a universal reagent for various schemes of biological microanalysis.

  3. Comparing compound-specific and bulk stable nitrogen isotope trophic discrimination factors across multiple freshwater fish species and diets

    Science.gov (United States)

    The use of nitrogen stable isotopes for estimation of animal trophic position has become an indispensable approach in food web ecology. Compound-specific isotope analysis of amino acids is a new approach for estimating trophic position that may overcome key issues associated with nitrogen stable iso...

  4. Ataluren and similar compounds (specific therapies for premature termination codon class I mutations) for cystic fibrosis.

    Science.gov (United States)

    Aslam, Aisha A; Higgins, Colin; Sinha, Ian P; Southern, Kevin W

    2017-01-19

    Cystic fibrosis is a common life-shortening genetic disorder in the Caucasian population (less common in other ethnic groups) caused by the mutation of a single gene that codes for the production of the cystic fibrosis transmembrane conductance regulator protein. This protein coordinates the transport of salt (and bicarbonate) across cell surfaces and the mutation most notably affects the airways. In the lungs of people with cystic fibrosis, defective protein results in a dehydrated surface liquid and compromised mucociliary clearance. The resulting thick mucus makes the airway prone to chronic infection and inflammation, which consequently damages the structure of the airways, eventually leading to respiratory failure. Additionally, abnormalities in the cystic fibrosis transmembrane conductance regulator protein lead to other systemic complications including malnutrition, diabetes and subfertility.Five classes of mutation have been described, depending on the impact of the mutation on the processing of the cystic fibrosis transmembrane conductance regulator protein in the cell. In class I mutations, the presence of premature termination codons prevents the production of any functional protein resulting in a severe cystic fibrosis phenotype. Advances in the understanding of the molecular genetics of cystic fibrosis has led to the development of novel mutation-specific therapies. Therapies targeting class I mutations (premature termination codons) aim to mask the abnormal gene sequence and enable the normal cellular mechanism to read through the mutation, potentially restoring the production of the cystic fibrosis transmembrane conductance regulator protein. This could in turn make salt transport in the cells function more normally and may decrease the chronic infection and inflammation that characterises lung disease in people with cystic fibrosis. To evaluate the benefits and harms of ataluren and similar compounds on clinically important outcomes in people with

  5. Distinguishing between Old and Modern Permafrost Sources with Compound-Specific Deuterium Analysis

    Science.gov (United States)

    Tesi, T.; Vonk, J.; Hugelius, G.; Kuhry, P.; Gustafsson, O.

    2015-12-01

    Permafrost underlies about a quarter of the northern hemispheric landmass and stores about 1450 Pg organic carbon (OC). When permafrost thaws, its OC is remobilized into the environment where it is available for degradation, transport or burial. The fate of this OC is partly determined by the type of thaw: (i) abrupt thaw of ice-rich Pleistocene permafrost causing collapse and deep exposure, and (ii) gradual thaw of the surface active layer causing release of topsoil, modern permafrost. The relative magnitude of each thaw pathway, along with its potential impacts on the permafrost carbon feedback, are however not known. We here present compound-specific deuterium analysis as a permafrost tracing tool to distinguish between old permafrost OC release, primarily released through abrupt thaw, and modern permafrost OC release, mostly released through gradual active layer thaw. We analyzed δ2H values of leaf waxes extracted from both Pleistocene-aged permafrost samples (n=9) and modern vegetation/O-horizon (topsoil) samples (n=9) from across northeast coastal Siberia. We analyzed both long-chain n-alkanes (chain lengths C25, C27, C29 and C31) as well as long-chain n-alkanoic acids (C22, C24, C26, C28). Results show more depleted values for the Pleistocene samples (-264±33 and -280±6.5‰ for average n-alkanes and n-alkanoic acids, respectively) than for the topsoil samples (-227±23 and -234±13‰). The variability in distribution and δ2H signals for n-alkanoic acids and n-alkanes of topsoils is higher than the Pleistocene samples, that show more constant values, yet the isotopic distinction between the two sources is consisten. For n-alkanes, there is a clear decrease in δ2H signal with chain length (from -201±32 to -247±23‰ for topsoil, and -221±26 to -297±15‰ for Pleistocene samples). These data suggest that δ2H analysis on leaf wax lipids in environments receiving OC from permafrost thaw can be used to assess the relative input of Pleistocene versus

  6. Authenticity of aroma components Enantiomeric separation and compound specific stable isotope analysis

    DEFF Research Database (Denmark)

    Hansen, Anne-Mette Sølvbjerg

    The word “authenticity” is increasingly used in the marketing of food products. A product can be marketed claiming its authenticity such as containing only natural ingredients or originating from a special location produced using local traditional production methods. Within the area of food...... ingredients a problem with authenticity of aroma compounds has occurred, because natural aromas are wholly or partly replaced with synthetic ones. This is a large economic problem, since natural aromas are often more expensive than artificial ones. Furthermore, the European Union has legal requirements...... necessary to develop new methods able to verify whether the authenticity of the food products is in agreement with the labelling. Some aroma compounds are chiral, meaning that they can exist as two enantiomers with different spatial orientation. Synthetic aroma compounds will consist of almost equal amounts...

  7. Degradation of specific aromatic compounds migrating from PEXpipes into drinking water

    DEFF Research Database (Denmark)

    Ryssel, Sune Thyge; Arvin, Erik; Lützhøft, Hans-Christian Holten

    2015-01-01

    extractions and spiked samples. 4-ethylphenol was degraded within 22 days. Eight compounds were, however, only partially degradable under abiotic and biotic conditions within the timeframe of the experiments (2e4 weeks). Neither inhibition nor co-metabolism was observed in the presence of acetate or PEX pipe...

  8. Evaluation of a liquid chromatography method for compound-specific δ13C analysis of plant carbohydrates in alkaline media.

    Science.gov (United States)

    Rinne, Katja T; Saurer, Matthias; Streit, Kathrin; Siegwolf, Rolf T W

    2012-09-30

    Isotope analysis of carbohydrates is important for improved understanding of plant carbon metabolism and plant physiological response to the environment. High-performance liquid chromatography/isotope ratio mass spectrometry (HPLC/IRMS) for direct compound-specific δ(13)C measurements of soluble carbohydrates has recently been developed, but the still challenging sample preparation and the fact that no single method is capable of separating all compounds of interest hinder its wide-spread application. Here we tested in detail a chromatography method in alkaline media. We examined the most suitable chromatographic conditions for HPLC/IRMS analysis of carbohydrates in aqueous conifer needle extracts using a CarboPac PA20 anion-exchange column with NaOH eluent, paying specific attention to compound yields, carbon isotope fractionation processes and the reproducibility of the method. Furthermore, we adapted and calibrated sample preparation methods for HPLC/IRMS analysis. OnGuard II cartridges were used for sample purification. Good peak separation and highly linear and reproducible concentration and δ(13)C measurements were obtained. The alkaline eluent was observed to induce isomerization of hexoses, detected as reduced yields and (13)C fractionation of the affected compounds. A reproducible pre-purification method providing ~100% yield for the carbohydrate compounds of interest was calibrated. The good level of peak separation obtained in this study is reflected in the good precision and linearity of concentration and δ(13)C results. The data provided crucial information on the behaviour of sugars in LC analysis with alkaline media. The observations highlight the importance for the application of compound-matched standard solution for the detection and correction of instrumental biases in concentration and δ(13)C analysis performed under identical chromatographic conditions. The calibrated pre-purification method is well suited for studies with complex matrices

  9. Tissue-Specific, Development-Dependent Phenolic Compounds Accumulation Profile and Gene Expression Pattern in Tea Plant [Camellia sinensis

    Science.gov (United States)

    Li, Weiwei; Zhao, Lei; Meng, Fei; Wang, Yunsheng; Tan, Huarong; Yang, Hua; Wei, Chaoling; Wan, Xiaochun; Gao, Liping; Xia, Tao

    2013-01-01

    Phenolic compounds in tea plant [Camellia sinensis (L.)] play a crucial role in dominating tea flavor and possess a number of key pharmacological benefits on human health. The present research aimed to study the profile of tissue-specific, development-dependent accumulation pattern of phenolic compounds in tea plant. A total of 50 phenolic compounds were identified qualitatively using liquid chromatography in tandem mass spectrometry technology. Of which 29 phenolic compounds were quantified based on their fragmentation behaviors. Most of the phenolic compounds were higher in the younger leaves than that in the stem and root, whereas the total amount of proanthocyanidins were unexpectedly higher in the root. The expression patterns of 63 structural and regulator genes involved in the shikimic acid, phenylpropanoid, and flavonoid pathways were analyzed by quantitative real-time polymerase chain reaction and cluster analysis. Based on the similarity of their expression patterns, the genes were classified into two main groups: C1 and C2; and the genes in group C1 had high relative expression level in the root or low in the bud and leaves. The expression patterns of genes in C2-2-1 and C2-2-2-1 groups were probably responsible for the development-dependent accumulation of phenolic compounds in the leaves. Enzymatic analysis suggested that the accumulation of catechins was influenced simultaneously by catabolism and anabolism. Further research is recommended to know the expression patterns of various genes and the reason for the variation in contents of different compounds in different growth stages and also in different organs. PMID:23646127

  10. Tissue-specific, development-dependent phenolic compounds accumulation profile and gene expression pattern in tea plant [Camellia sinensis].

    Science.gov (United States)

    Jiang, Xiaolan; Liu, Yajun; Li, Weiwei; Zhao, Lei; Meng, Fei; Wang, Yunsheng; Tan, Huarong; Yang, Hua; Wei, Chaoling; Wan, Xiaochun; Gao, Liping; Xia, Tao

    2013-01-01

    Phenolic compounds in tea plant [Camellia sinensis (L.)] play a crucial role in dominating tea flavor and possess a number of key pharmacological benefits on human health. The present research aimed to study the profile of tissue-specific, development-dependent accumulation pattern of phenolic compounds in tea plant. A total of 50 phenolic compounds were identified qualitatively using liquid chromatography in tandem mass spectrometry technology. Of which 29 phenolic compounds were quantified based on their fragmentation behaviors. Most of the phenolic compounds were higher in the younger leaves than that in the stem and root, whereas the total amount of proanthocyanidins were unexpectedly higher in the root. The expression patterns of 63 structural and regulator genes involved in the shikimic acid, phenylpropanoid, and flavonoid pathways were analyzed by quantitative real-time polymerase chain reaction and cluster analysis. Based on the similarity of their expression patterns, the genes were classified into two main groups: C1 and C2; and the genes in group C1 had high relative expression level in the root or low in the bud and leaves. The expression patterns of genes in C2-2-1 and C2-2-2-1 groups were probably responsible for the development-dependent accumulation of phenolic compounds in the leaves. Enzymatic analysis suggested that the accumulation of catechins was influenced simultaneously by catabolism and anabolism. Further research is recommended to know the expression patterns of various genes and the reason for the variation in contents of different compounds in different growth stages and also in different organs.

  11. Construction of target-specific virus-like particles for the delivery of algicidal compounds to harmful algae.

    Science.gov (United States)

    Kang, Beom Sik; Eom, Chi-Yong; Kim, Wonduck; Kim, Pyoung Il; Ju, Sun Yi; Ryu, Jaewon; Han, Gui Hwan; Oh, Jeong-Il; Cho, Hoon; Baek, Seung Ho; Kim, Gueeda; Kim, Minju; Hyun, Jaekyung; Jin, EonSeon; Kim, Si Wouk

    2015-04-01

    Harmful algal blooms (HABs) can lead to substantial socio-economic losses and extensive damage to aquatic ecosystems, drinking water sources and human health. Common algicidal techniques, including ozonation, ultrasonic treatment and dispersion of algae-killing chemicals, are unsatisfactory both economically and ecologically. This study therefore presents a novel alternative strategy for the efficient control of deleterious algae via the use of host-specific virus-like particles (VLPs) combined with chemically synthesized algicidal compounds. The capsid protein of HcRNAV34, a single-stranded RNA virus that infects the toxic dinoflagellate, Heterocapsa circularisquama, was expressed in and purified from Escherichia coli and then self-assembled into VLPs in vitro. Next, the algicidal compound, thiazolidinedione 49 (TD49), was encapsidated into HcRNAV34 VLPs for specific delivery to H. circularisquama. Consequently, HcRNAV34 VLPs demonstrated the same host selectivity as naturally occurring HcRNAV34 virions, while TD49-encapsidated VLPs showed a more potent target-specific algicidal effect than TD49 alone. These results indicate that target-specific VLPs for the delivery of cytotoxic compounds to nuisance algae might provide a safe, environmentally friendly approach for the management of HABs in aquatic ecosystems. © 2014 Society for Applied Microbiology and John Wiley & Sons Ltd.

  12. Elicitation of Induced Resistance against Pectobacterium carotovorum and Pseudomonas syringae by Specific Individual Compounds Derived from Native Korean Plant Species

    Directory of Open Access Journals (Sweden)

    Choong-Min Ryu

    2013-10-01

    Full Text Available Plants have developed general and specific defense mechanisms for protection against various enemies. Among the general defenses, induced resistance has distinct characteristics, such as broad-spectrum resistance and long-lasting effectiveness. This study evaluated over 500 specific chemical compounds derived from native Korean plant species to determine whether they triggered induced resistance against Pectobacterium carotovorum supsp. carotovorum (Pcc in tobacco (Nicotiana tabacum and Pseudomonas syringae pv. tomato (Pst in Arabidopsis thaliana. To select target compound(s with direct and indirect (volatile effects, a new Petri-dish-based in vitro disease assay system with four compartments was developed. The screening assay showed that capsaicin, fisetin hydrate, jaceosidin, and farnesiferol A reduced the disease severity significantly in tobacco. Of these four compounds, capsaicin and jaceosidin induced resistance against Pcc and Pst, which depended on both salicylic acid (SA and jasmonic acid (JA signaling, using Arabidopsis transgenic and mutant lines, including npr1 and NahG for SA signaling and jar1 for JA signaling. The upregulation of the PR2 and PDF1.2 genes after Pst challenge with capsaicin pre-treatment indicated that SA and JA signaling were primed. These results demonstrate that capsaicin and jaceosidin can be effective triggers of strong induced resistance against both necrotrophic and biotrophic plant pathogens.

  13. Experimental investigation of the impact of compound-specific dispersion and electrostatic interactions on transient transport and solute breakthrough

    DEFF Research Database (Denmark)

    Muniruzzaman, Muhammad; Rolle, Massimo

    2017-01-01

    This study investigates the effects of compound-specific diffusion/dispersion and electrochemical migration on transient solute transport in saturated porous media. We conducted laboratory bench-scale experiments, under advection-dominated regimes (seepage velocity: 0.5, 5, 25 m/d), in a quasi two....... The experimental results show that compound-specific effects and charge-induced Coulombic interactions are important not only at low velocities and/or for steady state plumes but also for transient transport under high flow velocities. Such effects can lead to a remarkably different behavior of measured...... breakthrough curves also at very high Péclet numbers. To quantitatively interpret the experimental results, we used four modeling approaches: classical advection-dispersion equation (ADE), continuous time random walk (CTRW), dual-domain mass transfer model (DDMT), and a multicomponent ionic dispersion model...

  14. Compound-specific stable carbon isotope ratios of phenols and nitrophenols derivatized with N,O-bis(trimethylsilyl)trifluoroacetamide

    Energy Technology Data Exchange (ETDEWEB)

    Irei, Satoshi, E-mail: irei.satoshi@nies.go.jp [Centre for Atmospheric Chemistry, Department of Chemistry, York University, 4700 Keele Street, Toronto, Ontario M3J 1P3 (Canada); Climate Research Division, Atmospheric Science and Technology Directorate, Science and Technology Branch, Environment Canada, 4905 Dufferin Street, Toronto, Ontario M3H 5T4 (Canada); Rudolph, Jochen [Centre for Atmospheric Chemistry, Department of Chemistry, York University, 4700 Keele Street, Toronto, Ontario M3J 1P3 (Canada); Huang, Lin [Climate Research Division, Atmospheric Science and Technology Directorate, Science and Technology Branch, Environment Canada, 4905 Dufferin Street, Toronto, Ontario M3H 5T4 (Canada)

    2013-07-05

    Graphical abstract: -- Highlights: •For GCC–IRMS analysis, an approach avoiding impact of NO{sub 2} on δ{sup 13}C was demonstrated. •Carbon isotope fractionations during derivatizing reactions here were negligible. •Except some labile compounds, the overall bias of the method here was −0.21‰. •Even for the labile compounds, measurement biases ranged +1.2‰ to −1.4‰. •Real sample analysis demonstrates usefulness of the method for fractionation study. -- Abstract: We developed an analytical method for measuring compound-specific stable carbon isotope ratios (δ{sup 13}C) of phenols and nitrophenols in filter samples of particulate organic matter. The method was tested on 13 phenols derivatized with N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA), together with four nonphenolic compounds. The data obtained by our method required two specific corrections for the determination of valid δ{sup 13}C values: (1) for nitro compounds, the routine correction with use of m/z 46 for the contribution of {sup 12}C{sup 17}O{sup 16}O molecules) to m/z 45 was modified due to impact of NO{sub 2} on the m/z 46 trace, and (2) for the derivatized phenols, measured δ{sup 13}C values were corrected for the shift in δ{sup 13}C due to the addition of carbon atoms from the BSTFA moiety. Analysis of standard-spiked filters showed that overall there was a small compound-dependent bias in the δ{sup 13}C values: the average bias ± the standard error of the mean of −0.21 ± 0.1‰ for the standard compounds tested, except 3-methylcatechol, methylhydroquinone, 4-methyl-2-nitrophenol, and 2,6-dimethyl-4-nitrophenol, whereas the average biases ± the standard errors of the mean for those were +1.2 ± 0.3‰, +1.2 ± 0.2‰, −1.2 ± 0.2‰, and −1.4 ± 0.5‰, respectively, when the injected mass of a derivatized compound exceeded 15 ngC. In situations where such small biases and uncertainties are acceptable, the method described here could be used to obtain valuable

  15. Specific heat of REBaSrCu{sub 3}O{sub 7} compounds (R=Nd, Sm, Dy, Er)

    Energy Technology Data Exchange (ETDEWEB)

    Tarnawski, Z. [Academy of Min. and Metall., Cracow (Poland). Solid State Phys. Dept.]|[Amsterdam Univ. (Netherlands). Van der Waals-Zeeman Inst.; Wang, X.Z. [Johannes Keppler Univ., Linz (Austria). Angewandte Physik; Kim-Ngan, N.-T.H. [Pedagogical Univ., Krakow (Poland). Center for Solid State Physics]|[Pedagogical Univ., Krakow (Poland). Inst. of Physics; Radwanski, R.J. [Pedagogical Univ., Krakow (Poland). Center for Solid State Physics]|[Pedagogical Univ., Krakow (Poland). Inst. of Physics; Franse, J.J.M. [Amsterdam Univ. (Netherlands). Van der Waals-Zeeman Inst.

    1996-05-01

    Specific heat measurements have been performed on polycrystalline REBaSrCu{sub 3}O{sub 7} (R=Nd, Sm, Dy, Er) in the temperature range from 0.3 to 3 K. Antiferromagnetic order is indicated by the {lambda}-type peak in the specific heat at 1.2 and 0.6 K for Dy and Er compounds, respectively, whereas only a broad anomaly around 0.6 K is observed for Nd and Sm compounds. The specific heat data for Nd and Sm compounds are well fit with a Schottky-type anomaly with an energy splitting of 1.4 K, indicating the existence of an internal field of 2.42 T at the RE site. The magnetic singlet ground state of ErBaSrCu{sub 3}O{sub 7} and DyBaSrCu{sub 3}O{sub 7} is produ ced by exchange interactions of -0.05 and -0.035 T f.u./{mu}{sub B}, respectively. (orig.).

  16. Analysis of common and specific mechanisms of liver function affected by nitrotoluene compounds.

    Directory of Open Access Journals (Sweden)

    Youping Deng

    Full Text Available BACKGROUND: Nitrotoluenes are widely used chemical manufacturing and munitions applications. This group of chemicals has been shown to cause a range of effects from anemia and hypercholesterolemia to testicular atrophy. We have examined the molecular and functional effects of five different, but structurally related, nitrotoluenes on using an integrative systems biology approach to gain insight into common and disparate mechanisms underlying effects caused by these chemicals. METHODOLOGY/PRINCIPAL FINDINGS: Sprague-Dawley female rats were exposed via gavage to one of five concentrations of one of five nitrotoluenes [2,4,6-trinitrotoluene (TNT, 2-amino-4,6-dinitrotoluene (2ADNT 4-amino-2,6-dinitrotoulene (4ADNT, 2,4-dinitrotoluene (2,4DNT and 2,6-dinitrotoluene (2,6DNT] with necropsy and tissue collection at 24 or 48 h. Gene expression profile results correlated well with clinical data and liver histopathology that lead to the concept that hematotoxicity was followed by hepatotoxicity. Overall, 2,4DNT, 2,6DNT and TNT had stronger effects than 2ADNT and 4ADNT. Common functional terms, gene expression patterns, pathways and networks were regulated across all nitrotoluenes. These pathways included NRF2-mediated oxidative stress response, aryl hydrocarbon receptor signaling, LPS/IL-1 mediated inhibition of RXR function, xenobiotic metabolism signaling and metabolism of xenobiotics by cytochrome P450. One biological process common to all compounds, lipid metabolism, was found to be impacted both at the transcriptional and lipid production level. CONCLUSIONS/SIGNIFICANCE: A systems biology strategy was used to identify biochemical pathways affected by five nitroaromatic compounds and to integrate data that tie biochemical alterations to pathological changes. An integrative graphical network model was constructed by combining genomic, gene pathway, lipidomic, and physiological endpoint results to better understand mechanisms of liver toxicity and

  17. Compound-specific effects of diverse neurodevelopmental toxicants on global gene expression in the neural embryonic stem cell test (ESTn)

    Energy Technology Data Exchange (ETDEWEB)

    Theunissen, P.T., E-mail: Peter.Theunissen@rivm.nl [Laboratory for Health Protection Research, National Institute for Public Health and the Environment (RIVM), Bilthoven (Netherlands); Department of Toxicogenomics, Maastricht University, Maastricht (Netherlands); Robinson, J.F. [Laboratory for Health Protection Research, National Institute for Public Health and the Environment (RIVM), Bilthoven (Netherlands); Department of Toxicogenomics, Maastricht University, Maastricht (Netherlands); Netherlands Toxicogenomics Centre, Maastricht (Netherlands); Pennings, J.L.A. [Laboratory for Health Protection Research, National Institute for Public Health and the Environment (RIVM), Bilthoven (Netherlands); Netherlands Toxicogenomics Centre, Maastricht (Netherlands); Herwijnen, M.H. van [Department of Toxicogenomics, Maastricht University, Maastricht (Netherlands); Kleinjans, J.C.S. [Department of Toxicogenomics, Maastricht University, Maastricht (Netherlands); Netherlands Toxicogenomics Centre, Maastricht (Netherlands); Piersma, A.H. [Laboratory for Health Protection Research, National Institute for Public Health and the Environment (RIVM), Bilthoven (Netherlands); Netherlands Toxicogenomics Centre, Maastricht (Netherlands); Institute for Risk Assessment Sciences, Faculty of Veterinary Sciences, Utrecht University, Utrecht (Netherlands)

    2012-08-01

    Alternative assays for developmental toxicity testing are needed to reduce animal use in regulatory toxicology. The in vitro murine neural embryonic stem cell test (ESTn) was designed as an alternative for neurodevelopmental toxicity testing. The integration of toxicogenomic-based approaches may further increase predictivity as well as provide insight into underlying mechanisms of developmental toxicity. In the present study, we investigated concentration-dependent effects of six mechanistically diverse compounds, acetaldehyde (ACE), carbamazepine (CBZ), flusilazole (FLU), monoethylhexyl phthalate (MEHP), penicillin G (PENG) and phenytoin (PHE), on the transcriptome and neural differentiation in the ESTn. All compounds with the exception of PENG altered ESTn morphology (cytotoxicity and neural differentiation) in a concentration-dependent manner. Compound induced gene expression changes and corresponding enriched gene ontology biological processes (GO–BP) were identified after 24 h exposure at equipotent differentiation-inhibiting concentrations of the compounds. Both compound-specific and common gene expression changes were observed between subsets of tested compounds, in terms of significance, magnitude of regulation and functionality. For example, ACE, CBZ and FLU induced robust changes in number of significantly altered genes (≥ 687 genes) as well as a variety of GO–BP, as compared to MEHP, PHE and PENG (≤ 55 genes with no significant changes in GO–BP observed). Genes associated with developmentally related processes (embryonic morphogenesis, neuron differentiation, and Wnt signaling) showed diverse regulation after exposure to ACE, CBZ and FLU. In addition, gene expression and GO–BP enrichment showed concentration dependence, allowing discrimination of non-toxic versus toxic concentrations on the basis of transcriptomics. This information may be used to define adaptive versus toxic responses at the transcriptome level.

  18. Effects of thermal maturation and thermochemical sulfate reduction on compound-specific sulfur isotopic compositions of organosulfur compounds in Phosphoria oils from the Bighorn Basin, USA

    Science.gov (United States)

    Ellis, Geoffrey S.; Said-Ahamed, Ward; Lillis, Paul G.; Shawar, Lubna; Amrani, Alon

    2017-01-01

    Compound-specific sulfur isotope analysis was applied to a suite of 18 crude oils generated from the Permian Phosphoria Formation in the Bighorn Basin, western USA. These oils were generated at various levels of thermal maturity and some experienced thermochemical sulfate reduction (TSR). This is the first study to examine the effects of thermal maturation on stable sulfur isotopic compositions of individual organosulfur compounds (OSCs) in crude oil. A general trend of 34S enrichment in all of the studied compounds with increasing thermal maturity was observed, with the δ34S values of alkyl-benzothiophenes (BTs) tending to be enriched in 34S relative to those of the alkyl-dibenzothiophenes (DBTs) in lower-maturity oils. As thermal maturity increases, δ34S values of both BTs and DBTs become progressively heavier, but the difference in the average δ34S value of the BTs and DBTs (Δ34S BT-DBT) decreases. Differences in the isotopic response to thermal stress exhibited by these two compound classes are considered to be the result of relative differences in their thermal stabilities. TSR-altered Bighorn Basin oils have OSCs that are generally enriched in 34S relative to non-TSR-altered oils, with the BTs being enriched in 34S relative to the DBTs, similar to the findings of previous studies. However, several oils that were previously interpreted to have been exposed to minor TSR have Δ34S BT-DBT values that do not support this interpretation. The δ34S values of the BTs and DBTs in some of these oils suggest that they did not experience TSR, but were derived from a more thermally mature source. The heaviest δ34S values observed in the OSCs are enriched in 34S by up to 10‰ relative to that of Permian anhydrite in the Bighorn Basin, suggesting that there may be an alternate or additional source of sulfate in some parts of the basin. These results indicate that the sulfur isotopic composition of OSCs in oil provides a sensitive indicator for the extent of TSR

  19. Study of thermochemical sulfate reduction mechanism using compound specific sulfur isotope analysis

    Science.gov (United States)

    Meshoulam, Alexander; Ellis, Geoffrey S.; Ahmad, Ward Said; Deev, Andrei; Sessions, Alex L.; Tang, Yongchun; Adkins, Jess F.; Liu, Jinzhong; Gilhooly, William P.; Aizenshtat, Zeev; Amrani, Alon

    2016-01-01

    The sulfur isotopic fractionation associated with the formation of organic sulfur compounds (OSCs) during thermochemical sulfate reduction (TSR) was studied using gold-tube pyrolysis experiments to simulate TSR. The reactants used included n-hexadecane (n-C16) as a model organic compound with sulfate, sulfite, or elemental sulfur as the sulfur source. At the end of each experiment, the S-isotopic composition and concentration of remaining sulfate, H2S, benzothiophene, dibenzothiophene, and 2-phenylthiophene (PT) were measured. The observed S-isotopic fractionations between sulfate and BT, DBT, and H2S in experimental simulations of TSR correlate well with a multi-stage model of the overall TSR process. Large kinetic isotope fractionations occur during the first, uncatalyzed stage of TSR, 12.4‰ for H2S and as much as 22.2‰ for BT. The fractionations decrease as the H2S concentration increases and the reaction enters the second, catalyzed stage. Once all of the oxidizable hydrocarbons have been consumed, sulfate reduction ceases and equilibrium partitioning then dictates the fractionation between H2S and sulfate (∼17‰).

  20. Species-specific accumulation of dioxin related compounds in cetaceans collected from Japanese coastal waters

    Energy Technology Data Exchange (ETDEWEB)

    Kajiwara, N.; Watanabe, M.; Tanabe, S. [Center for Marine Environmental Studies (CMES), Ehime Univ. (Japan); Amano, M. [Ocean Research Inst., Univ. of Tokyo, Iwate (Japan); Yamada, T. [National Science Museum, Tokyo (Japan)

    2004-09-15

    Polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) are extremely hazardous and persistent chemicals identified as contaminants in chlorophenols, herbicides, fly ash and other incineration products. Dioxin-like PCBs including non- and mono-ortho coplanar PCBs are referred to as dioxin related compounds and are evaluated on par with PCDD/Fs in environmental risks since they have a high toxicity, similar to that of PCDD/Fs. These congeners have a range of physicochemical characteristics, which profoundly affect their persistence, environmental distribution, and bioaccumulation in aquatic food chains. Fish-eating wildlife such as marine mammals are particularly vulnerable to such contamination given their long lives, high trophic level, relative inability to metabolize many persistent organic pollutants (POPs), and the biomagnification of these contaminants in aquatic food chains. However, most studies dealing with PCDDs and PCDFs in marine mammals have been carried out on pinnipeds, and data on PCDD/Fs levels in cetaceans are scarce. The present study is aimed at understanding the recent pattern of contamination by dioxin related compounds including non- and mono-ortho coplanar PCBs and PCDD/Fs in three cetacean species collected from Japanese coastal waters during 1998-2001, and also to discuss the factors determining the accumulation.

  1. Non-targeted metabolite profiling highlights the potential of strawberry leaves as a resource for specific bioactive compounds.

    Science.gov (United States)

    Kårlund, Anna; Hanhineva, Kati; Lehtonen, Marko; McDougall, Gordon J; Stewart, Derek; Karjalainen, Reijo O

    2017-05-01

    The non-edible parts of horticultural crops, such as leaves, contain substantial amounts of valuable bioactive compounds which are currently only little exploited. For example, strawberry (Fragaria × ananassa) leaves may be a promising bioresource for diverse health-related applications. However, product standardization sets a real challenge, especially when the leaf material comes from varying cultivars. The first step towards better quality control of berry fruit leaf-based ingredients and supplements is to understand metabolites present and their stability in different plant cultivars, so this study surveyed the distribution of potentially bioactive strawberry leaf metabolites in six different strawberry cultivars. Non-targeted metabolite profiling analysis using LC/qTOF-ESI-MS with data processing via principal component analysis and k-means clustering analysis was utilized to examine differences and commonalities between the leaf metabolite profiles. Quercetin and kaempferol derivatives were the dominant flavonol groups in strawberry leaves. Previously described and novel caffeic and chlorogenic acid derivatives were among the major phenolic acids. In addition, ellagitannins were one of the distinguishing compound classes in strawberry leaves. In general, strawberry leaves also contained high levels of octadecatrienoic acid derivatives, precursors of valuable odour compounds. The specific bioactive compounds found in the leaves of different strawberry cultivars offer the potential for the selection of optimized leaf materials for added-value food and non-food applications. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  2. Interpretation of compound nouns by adolescents with specific language impairment and autism spectrum disorders: an investigation of phenotypic overlap.

    Science.gov (United States)

    Riches, Nick G; Loucas, Tom; Baird, Gillian; Charman, Tony; Simonoff, Emily

    2012-08-01

    The study aimed to investigate (i) whether adolescents with Specific Language Impairment (SLI) and Autism plus Language Impairment (ALI) experience word-formation difficulties, and (ii) whether these two groups present with a similar language phenotype. The study investigated four groups using a 2 (language status) ×2 (autism status) design; adolescents with SLI (n = 14), ALI (n = 16), Autism Language-Typical (ALT; n = 14), and language matched controls (n = 17), with all groups presenting with typical non-verbal skills. Mean age was 14;10. Comprehension of conventional Noun-Noun lexical compounds (e.g., snowman), synthetic compounds (SCs, e.g., cat chaser), and novel root compounds (RCs, e.g., sheep socks), was assessed using a forced-choice picture selection task. The SLI and ALI participants frequently mis-parsed the SCs, interpreting the first noun as the agent. Those with poorer vocabularies and non-word repetition had greater difficulties. Reaction time (RT) profiles were flatter in the ASD groups, with similar RTs across different compounds. Language difficulties in the SLI and ALI groups extend to word-formation processes; for example, comprehension of SCs. This may reflect difficulties making analogies with stored lexical items. Overall the results support the hypothesis of a phenotypic overlap between SLI and ALI.

  3. Environmental Forensics : Compound Specific Isotope Analysis Of PAHs. Study Of A Former Coal Tar Plant.

    Science.gov (United States)

    Assal, A.; Doherty, R.; Dickson, K.; Kalin, R. M.

    2008-12-01

    Stable carbon isotopic fingerprints of PAHs obtained by GC-IRMS have often been used in source apportionment studies. The use of PAHs in environmental forensics relies on the assumption that carbon isotopic fractionation caused by microbial degradation is less significant for these heavy molecular weight compounds than for lighter molecules such as chlorinated solvents or BTEX. Carbon isotopic fractionation of PAHs during degradation is still not well understood. The aim of this study was to assess the potential of CSIA of PAHs for environmental forensics applications at a complex (hydrogeology affected by tidal fluxes) former coal tar plant. In this work, soil samples from a tar works site were analyzed. The tar works operated on the site over a period of sixty years. A source apportionment study was first carried out based on 90 target PAHs quantified by GC-MS. These results were then compared to carbon isotope fingerprints. The separation of compounds of interest from co-extracted interfering peaks is a crucial prerequisite of CSIA by GC-IRMS. Hence, a sample preparation method which allowed the determination of precise carbon isotope signatures for up to 35 compounds per soil extract was developed, validated and applied to the samples previously analyzed by GC- MS. Although most soil samples were shown to be related to the point source tar contamination, PAHs ratios and principal component analysis of abundances highlighted some samples with unusual patterns, suggesting the input of a second source of contaminants. However, no statistically significant variation of the isotopic fingerprints of heavy molecular weight PAHs of these samples was observed. This was inconsistent with the first diagnosis. Since evidence was provided that most samples were only affected by a single source of contaminants, carbon isotopic fractionation was investigated in-situ. Importantly, naphthalene and 2- and 1- methylnaphthalenes isotopic fractionation was observed in a vertical

  4. Microbial utilization of amino acids in soil assessed by position-specific labeling and compound-specific 13C-PLFA-analysis

    Science.gov (United States)

    Dippold, M.; Apostel, C.; Glaser, B.; Kuzyakov, Y.

    2012-04-01

    Transformation of low molecular weight organic substances (LMWOS) organic substances in soil is one of the most important processes in the turnover of organic matter, as all high molecular substances pass this stage during their decomposition. Microbial utilization is the most important process for the transformation of LMWOS in soil and thus is an important process in the turnover of organic matter. Position-specific labeling combined with compound-specific 13C analysis of microbial biomass allows a closer look on the mechanisms of LMWOS transformation in soil. We assessed short-term (3 and 10 days) transformations of the amino acids by adding position-specifically labeled 13C and 15N alanine and glutamic acid to soil in a field experiment. We quantified the microbial utilization of the different functional groups by 13C- and 15N-analysis of microbial biomass with the chloroform-fumigation-extraction method. A more specific look on the utilization of individual C positions by distinct microbial groups was gained by 13C-PLFA analysis. Microbial degradation was fastest with the highly oxidized carboxylic groups of the amino acids, whereas more reduced C positions showed a higher utilization by microbial anabolism. Microbial groups revealed different incorporation of specific C positions into their PLFA. The highest incorporation was reached by the prokaryotic groups, especially the gram-negatives. Whereas alanine was metabolized similar by different microbial groups glutamic acid C showed different utilization of the individual C positions for distinct microbial groups. This may arise from the use of glutamic acid as a general N transport molecule in bacterial metabolism. Hence, position-specifically labeled glutamic acid may act as a tracer of bacterial N metabolism. The application of position-specifically labeled substances opens a new way to investigate the microbial transformations of amino acids in soil. Observing single C atoms and their utilization by

  5. Compound Specific Extraction of Camptothecin from Nothapodytes nimmoniana and Piperine from Piper nigrum Using Accelerated Solvent Extractor.

    Science.gov (United States)

    Upadhya, Vinayak; Pai, Sandeep R; Sharma, Ajay K; Hegde, Harsha V; Kholkute, Sanjiva D; Joshi, Rajesh K

    2014-01-01

    Effects of varying temperatures with constant pressure of solvent on extraction efficiency of two chemically different alkaloids were studied. Camptothecin (CPT) from stem of Nothapodytes nimmoniana (Grah.) Mabb. and piperine from the fruits of Piper nigrum L. were extracted using Accelerated Solvent Extractor (ASE). Three cycles of extraction for a particular sample cell at a given temperature assured complete extraction. CPT and piperine were determined and quantified by using a simple and efficient UFLC-PDA (245 and 343 nm) method. Temperature increased efficiency of extraction to yield higher amount of CPT, whereas temperature had diminutive effect on yield of piperine. Maximum yield for CPT was achieved at 80°C and for piperine at 40°C. Thus, the study determines compound specific extraction of CPT from N. nimmoniana and piperine from P. nigrum using ASE method. The present study indicates the use of this method for simple, fast, and accurate extraction of the compound of interest.

  6. Compound-specific stable carbon isotopic signature of carbohydrate pyrolysis products from C3 and C4 plants.

    Science.gov (United States)

    González-Pérez, José A; Jiménez-Morillo, Nicasio T; de la Rosa, José M; Almendros, Gonzalo; González-Vila, Francisco J

    2016-02-01

    Pyrolysis-compound specific isotopic analysis (Py-CSIA: Py-GC-(FID)-C-IRMS) is a relatively novel technique that allows on-line quantification of stable isotope proportions in chromatographically separated products released by pyrolysis. Validation of the Py-CSIA technique is compulsory for molecular traceability in basic and applied research. In this work, commercial sucrose from C4 (sugarcane) and C3 (sugarbeet) photosystem plants and admixtures were studied using analytical pyrolysis (Py-GC/MS), bulk δ(13)C IRMS and δ(13)C Py-CSIA. Major pyrolysis compounds were furfural (F), furfural-5-hydroxymethyl (HMF) and levoglucosan (LV). Bulk and main pyrolysis compound δ(13)C (‰) values were dependent on plant origin: C3 (F, -24.65 ± 0.89; HMF, -22.07 ± 0.41‰; LV, -21.74 ± 0.17‰) and C4 (F, -14.35 ± 0.89‰; HMF, -11.22 ± 0.54‰; LV, -11.44 ± 1.26‰). Significant regressions were obtained for δ(13)C of bulk and pyrolysis compounds in C3 and C4 admixtures. Furfural (F) was found (13)C depleted with respect to bulk and HMF and LV, indicating the incorporation of the light carbon atom in position 6 of carbohydrates in the furan ring after pyrolysis. This is the first detailed report on the δ(13)C signature of major pyrolytically generated carbohydrate-derived molecules. The information provided by Py-CSIA is valuable for identifying source marker compounds of use in food science/fraud detection or in environmental research. © 2015 Society of Chemical Industry.

  7. Study of thermochemical sulfate reduction mechanism using compound specific sulfur isotope analysis

    Science.gov (United States)

    Meshoulam, Alexander; Ellis, Geoffrey S.; Said Ahmad, Ward; Deev, Andrei; Sessions, Alex L.; Tang, Yongchun; Adkins, Jess F.; Liu, Jinzhong; Gilhooly, William P.; Aizenshtat, Zeev; Amrani, Alon

    2016-09-01

    The sulfur isotopic fractionation associated with the formation of organic sulfur compounds (OSCs) during thermochemical sulfate reduction (TSR) was studied using gold-tube pyrolysis experiments to simulate TSR. The reactants used included n-hexadecane (n-C16) as a model organic compound with sulfate, sulfite, or elemental sulfur as the sulfur source. At the end of each experiment, the S-isotopic composition and concentration of remaining sulfate, H2S, benzothiophene, dibenzothiophene, and 2-phenylthiophene (PT) were measured. The observed S-isotopic fractionations between sulfate and BT, DBT, and H2S in experimental simulations of TSR correlate well with a multi-stage model of the overall TSR process. Large kinetic isotope fractionations occur during the first, uncatalyzed stage of TSR, 12.4‰ for H2S and as much as 22.2‰ for BT. The fractionations decrease as the H2S concentration increases and the reaction enters the second, catalyzed stage. Once all of the oxidizable hydrocarbons have been consumed, sulfate reduction ceases and equilibrium partitioning then dictates the fractionation between H2S and sulfate (∼17‰). Experiments involving sparingly soluble CaSO4 show that during the second catalytic phase of TSR the rate of sulfate reduction exceeds that of sulfate dissolution. In this case, there is no apparent isotopic fractionation between source sulfate and generated H2S, as all of the available sulfate is effectively reduced at all reaction times. When CaSO4 is replaced with fully soluble Na2SO4, sulfate dissolution is no longer rate limiting and significant S-isotopic fractionation is observed. This supports the notion that CaSO4 dissolution can lead to the apparent lack of fractionation between H2S and sulfate produced by TSR in nature. The S-isotopic composition of individual OSCs record information related to geochemical reactions that cannot be discerned from the δ34S values obtained from bulk phases such as H2S, oil, and sulfate minerals, and

  8. Chlorogenic acid was specifically induced among phenolic compounds in centipedegrass by gamma irradiation

    Energy Technology Data Exchange (ETDEWEB)

    An, Byung Chul; Barampuram, Shyamkumar; Lee, Seung Sik; Lee, Eun Mi; Chung, Byung Yeoup [Korea Atomic Energy Research Institute, Jeongeup (Korea, Republic of)

    2010-03-15

    Centipedegrass is a warm season turfgrass in the world. Chlorogenic acid (CA) is one of the important compounds present in the leaf of centipedegrass and already known as an antioxidant, CA has become a key resistance against insect pests and bacteria pathogens of agricultural and horticultural plants during seedlling stage. Furthermore, CA is accumulated by abiotic stress such as an UV irradiation. In present study, we investigated enhancement of the level of CA upon gamma irradiation in centipedegrass. The high performance liquid chromatography (HPLC) data analysis showed an approximately increasing of the CA levels from among the irradiated samples. However, plants irradiated at 50 Gy showed a constant increase in the CA level (0.0066 to 0.114 mg ml{sup -1} and 0.0258 to 0.2211 mg ml{sup -1}, respectively) from 3{sup rd} to 15{sup th} day among one and three month irradiated plants compared to control. The present study, indicates an increase in the CA level upon gamma irradiation, suggests strategy for conferment of strong resistance on seedling stage plants by gamma irradiation as simplicity and cheaply method.

  9. [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

  10. Compound Specific Hydrogen Isotope Composition of Type II and III Kerogen Extracted by Pyrolysis-GC-MS-IRMS

    Science.gov (United States)

    Socki, Richard A.; Pernia, Denet; Evans, Michael; Fu, Qi; Bissada, Kadry K.; Curiale, Joseph A.; Niles, Paul B.

    2013-01-01

    The use of Hydrogen (H) isotopes in understanding oil and gas resource plays is in its infancy. Described here is a technique for H isotope analysis of organic compounds pyrolyzed from oil and gas shale-derived kerogen. Application of this technique will progress our understanding. This work complements that of Pernia et al. (2013, this meeting) by providing a novel method for the H isotope analysis of specific compounds in the characterization of kerogen extracted by analytically diverse techniques. Hydrogen isotope analyses were carried out entirely "on-line" utilizing a CDS 5000 Pyroprobe connected to a Thermo Trace GC Ultra interfaced with a Thermo MAT 253 IRMS. Also, a split of GC-separated products was sent to a DSQ II quadrupole MS to make semi-quantitative compositional measurements of the extracted compounds. Kerogen samples from five different basins (type II and III) were dehydrated (heated to 80 C overnight in vacuum) and analyzed for their H isotope compositions by Pyrolysis-GC-MS-TC-IRMS. This technique takes pyrolysis products separated via GC and reacts them in a high temperature conversion furnace (1450 C) which quantitatively forms H2, following a modified method of Burgoyne and Hayes, (1998, Anal. Chem., 70, 5136-5141). Samples ranging from approximately 0.5 to 1.0mg in size, were pyrolyzed at 800 C for 30s. Compounds were separated on a Poraplot Q GC column. Hydrogen isotope data from all kerogen samples typically show enrichment in D from low to high molecular weight compounds. Water (H2O) average deltaD = -215.2 (V-SMOW), ranging from -271.8 for the Marcellus Shale to -51.9 for the Polish Shale. Higher molecular weight compounds like toluene (C7H8) have an average deltaD of -89.7 0/00, ranging from -156.0 for the Barnett Shale to -50.0 for the Monterey Shale. We interpret these data as representative of potential H isotope exchange between hydrocarbons and sediment pore water during formation within each basin. Since hydrocarbon H isotopes

  11. Oral Mucocutaneous Lesions – A Comparative Clinicopathological and Immunofluorescence Study

    Science.gov (United States)

    Rameshkumar, Annasamy; Varghese, Alex Kumaranthara; Dineshkumar, Thayalan; Ahmed, Shaheen; Venkatramani, Janaki; Sugirtharaj, G

    2015-01-01

    Background: Oral mucosa is often affected by many diseases including mucocutaneous disorders. The diagnoses of these disorders are primarily based on history, clinical features, and histopathology. For the past few years’ immunofluorescence techniques emerged as an important tool to study the pathogenesis and in the diagnosis of oral mucocutaneous and vesiculobullous disorders. The present study was designed to carry out retrospective and prospective analysis of oral mucocutaneous lesions to elucidate the clinicopathologic features and its immunofluorescence findings. Materials and Methods: A total of 70 subjects with oral mucocutaneous lesions were retrieved from the oral pathology files of Tamil Nadu Govt. Dental College and their clinical features were evaluated, and the histopathology was also evaluated with the help of hematoxylin and eosin stained sections. For the prospective study, biopsy from 12 subjects with oral mucocutaneous lesions was subjected to routine histopathological examination and DIF to evaluate the consistency of the methods. Results: In the retrospective analysis of 70 subjects with oral mucocutaneous lesions in relation to clinical features and histopathology, most of the findings were similar to the previous studies except for few criteria like male predilection in lichen planus and mucous membrane pemphigoid (MMP) and more prevalence of pemphigus vulgaris than MMP (2:1). In the prospective analysis of 12 subjects, the histopathological diagnosis was consistent with DIF study in 66% of cases. Conclusion: The diagnostic efficiency of oral mucocutaneous lesions can be improved by modern tools like DIF studies in addition to traditional methods like clinical and histopathology. PMID:25878481

  12. 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.

  13. Durability of direct immunofluorescence (DIF) slides stored at room temperature.

    Science.gov (United States)

    Elbendary, Amira; Zhou, Cheng; Truong, Jonathan; Elston, Dirk M

    2015-12-01

    Prior studies suggested that direct immunofluorescence (DIF) slides can be stored at room temperature. We sought to determine the durability of DIF slides stored at room temperature for 5 years. This was a retrospective study of 83 DIF slides archived at room temperature during 2010. The pattern of immunoreactants was compared with those noted in the original report. Loss of reactivity was limited to cases with weak fluorescence at original diagnosis. Loss of IgG was noted in 12.5% of cases, IgA in 12%, C3 in 10%, and IgM in 9.75%. Fibrin showed no loss of reactivity. Preservation of immunofluorescence was not related to site of deposition. Overall, a reliable diagnosis could be made in 75 of 79 archived cases (94.9%). Cases had been archived for periods varying from 4.5 to 5 years. Variations in processing and fluorochromes could affect durability. We have no way of knowing how long slides had been exposed to ultraviolet light at the time of initial examination. DIF showed excellent durability in slides kept at room temperature for 5 years. Copyright © 2015 American Academy of Dermatology, Inc. Published by Elsevier Inc. All rights reserved.

  14. Evaluation of two immunofluorescence assays with monoclonal antibodies for typing of herpes simplex virus.

    OpenAIRE

    Swierkosz, E M; Arens, M Q; Schmidt, R R; Armstrong, T

    1985-01-01

    An indirect immunofluorescence assay and a direct immunofluorescence assay were evaluated for typing clinical isolates of herpes simplex virus (HSV). The indirect immunofluorescence assay (Electro-Nucleonics, Inc.) correctly identified 16 HSV type 2 (HSV-2) isolates, but failed to identify 4 of 14 HSV-1 isolates because of background fluorescence and instability of reagents. Forty-nine HSV-1 isolates were correctly typed by direct immunofluorescence assay (Kallestad Laboratories, Inc.), but 1...

  15. Country-specific chemical signatures of persistent environmental compounds in breast milk

    DEFF Research Database (Denmark)

    Krysiak-Baltyn, Konrad; Toppari, J.; Skakkebaek, N.E.

    2010-01-01

    Recent reports have confirmed a worldwide increasing trend of testicular cancer incidence, and a conspicuously high prevalence of this disease and other male reproductive disorders, including cryptorchidism and hypospadias, in Denmark. In contrast, Finland, a similarly industrialized Nordic country......, exhibits much lower incidences of these disorders. The reasons behind the observed trends are unexplained, but environmental endocrine disrupting chemicals (EDCs) that affect foetal testis development are probably involved. Levels of persistent chemicals in breast milk can be considered a proxy...... to compare exposure of mothers to this environmental mixture of EDCs. Using sophisticated, bioinformatic tools in our analysis, we reveal, for the first time, distinct country-specific chemical signatures of EDCs with Danes having generally higher exposure than Finns to persistent bioaccumulative chemicals...

  16. Ammonium improves elution of fixed dried blood spots without affecting immunofluorescence assay quality.

    Science.gov (United States)

    Borremans, Benny

    2014-04-01

    To solve the problem of fixed dried blood spot elution without damaging IgG antibodies. The minimum effective concentration of liquid ammonium (NH3 ) in a PBS solution, which was found to elute fixed blood, was determined. By using a dilution series, the effects of NH3 on IgG antibody quality were assessed using immunofluorescence assays. The minimum effective concentration of 0.2% NH3 has no detectable effects on IgG quality. Ammonium greatly improves blood elution from fixed DBS while maintaining IgG antibody quality. These results are encouraging and provide a basis for further testing of the efficacy of ammonium in different situations as well as its effect on other compounds. © 2014 John Wiley & Sons Ltd.

  17. V. Detection of the Virus in Infected Materials by Immunofluorescence

    Science.gov (United States)

    Ruckerbauer, G. M.; Gray, D. P.; Girard, A.; Bannister, G. L.; Boulanger, P.

    1967-01-01

    The fluorescent-antibody technique was employed for the detection of bluetongue virus in bovine foetal kidney cell cultures inoculated with tissues and blood from experimentally infected animals. In the first series, a total number of 79 inoculated suckling-mouse brains were examined, 36 as frozen sections alone and 43 as impression slides in conjunction with tissue culture inoculation of the same material. With the combined tissue culture immunofluorescent methods, 36 suspicious were detected by impression smears and 37 positives by the tissue culture out of 43 brains examined. Twenty-two were suspicious out of the 36 examined as frozen sections. Results obtained with the second series, using sheep tissues, showed that the combined tissue culture-fluorescent antibody method was satisfactory for demonstrating the virus in blood of infected animals 1 to 9 days postinfection, and in some organs after death. No false positive reactions were obtained. ImagesFig. 1. PMID:4227047

  18. Compound-specific delta18O analyses of neutral sugars in soils using GC-Py-IRMS

    Science.gov (United States)

    Zech, Michael; Glaser, Bruno; Werner, Roland; Juchelka, Dieter

    2010-05-01

    Stable oxygen isotopes (delta18O) are a valuable (paleo-)climate proxy applied to ice cores, deep sea sediments, speleothems and lake sediments. For instance, there has been made much effort to assess delta18O of cellulose from lake sediments. However, purity grade of the extracted cellulose and hygroscopy are analytical problems. In order to overcome these problems, we tested compound-specific delta18O analyses of neutral sugars using gas chromatography - pyrolysis - isotope ratio mass spectrometry (GC-Py-IRMS). Although this technique is available for about the last ten years, it is hardly applied so far. We ensured quantitative conversion of organically bound oxygen to CO in the pyrolysis reactor by monitoring not only mass 28 but also mass 44 (CO2) and excluded oxygen contamination in our system by co-analysing oxygen-free compounds (alkanes). Neutral sugars from litter and topsoils were extracted hydrolytically and measured after derivatization with methylboronic acid (MBA) and N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA). Corrections were made using sugar standards measured in alternation with the samples to ensure the ‘principle of identical treatment' and for the hydrolytically introduced and exchangeable carbonyl-oxygen bound to C1 in the alditose sugar molecules. First results are promising for the hemicellulose-derived sugars arabinose and xylose as well as for the microbial-derived sugars fucose and rhamnose. Ongoing work focuses on assessing also cellulose-derived glucose and on the analytical elimination of the carbonyl-oxygen of the sugars.

  19. Compound Specific Extraction of Camptothecin from Nothapodytes nimmoniana and Piperine from Piper nigrum Using Accelerated Solvent Extractor

    Directory of Open Access Journals (Sweden)

    Vinayak Upadhya

    2014-01-01

    Full Text Available Effects of varying temperatures with constant pressure of solvent on extraction efficiency of two chemically different alkaloids were studied. Camptothecin (CPT from stem of Nothapodytes nimmoniana (Grah. Mabb. and piperine from the fruits of Piper nigrum L. were extracted using Accelerated Solvent Extractor (ASE. Three cycles of extraction for a particular sample cell at a given temperature assured complete extraction. CPT and piperine were determined and quantified by using a simple and efficient UFLC-PDA (245 and 343 nm method. Temperature increased efficiency of extraction to yield higher amount of CPT, whereas temperature had diminutive effect on yield of piperine. Maximum yield for CPT was achieved at 80∘C and for piperine at 40∘C. Thus, the study determines compound specific extraction of CPT from N. nimmoniana and piperine from P. nigrum using ASE method. The present study indicates the use of this method for simple, fast, and accurate extraction of the compound of interest.

  20. Experimental investigation of the impact of compound-specific dispersion and electrostatic interactions on transient transport and solute breakthrough

    Science.gov (United States)

    Muniruzzaman, Muhammad; Rolle, Massimo

    2017-02-01

    This study investigates the effects of compound-specific diffusion/dispersion and electrochemical migration on transient solute transport in saturated porous media. We conducted laboratory bench-scale experiments, under advection-dominated regimes (seepage velocity: 0.5, 5, 25 m/d), in a quasi two-dimensional flow-through setup using pulse injection of multiple tracers (both uncharged and ionic species). Extensive sampling and measurement of solutes' concentrations (˜1500 samples; >3000 measurements) were performed at the outlet of the flow-through setup, at high spatial and temporal resolution. The experimental results show that compound-specific effects and charge-induced Coulombic interactions are important not only at low velocities and/or for steady state plumes but also for transient transport under high flow velocities. Such effects can lead to a remarkably different behavior of measured breakthrough curves also at very high Péclet numbers. To quantitatively interpret the experimental results, we used four modeling approaches: classical advection-dispersion equation (ADE), continuous time random walk (CTRW), dual-domain mass transfer model (DDMT), and a multicomponent ionic dispersion model. The latter is based on the multicomponent formulation of coupled diffusive/dispersive fluxes and was used to describe and explain the electrostatic effects of charged species. Furthermore, we determined experimentally the temporal profiles of the flux-related dilution index. This metric of mixing, used in connection with the traditional solute breakthrough curves, proved to be useful to correctly distinguish between plume spreading and mixing, particularly for the cases in which the sole analysis of integrated concentration breakthrough curves may lead to erroneous interpretation of plume dilution.

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

    Directory of Open Access Journals (Sweden)

    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.

  3. Use of immunofluorescence technique in cultured fibroblasts from Mediterranean cetaceans as new "in vitro" tool to investigate effects of environmental contaminants.

    Science.gov (United States)

    Marsili, Letizia; Casini, Silvia; Bucalossi, Daniela; Porcelloni, Serena; Maltese, Silvia; Fossi, Maria Cristina

    2008-07-01

    The aim of the present study was to propose the immunofluorescence technique in cultured fibroblasts from Mediterranean cetaceans as a new "in vitro" tool to explore the susceptibility of these marine mammals to different xenobiotic compounds. The cell lines were cultured from integument biopsies of free-ranging and stranded cetaceans (dead within 12h). Using the indirect immunofluorescence assay, we detected endogenous proteins induced by different contaminants. Here we present the method used for qualitative and quantitative evaluation of cytochromes P450 (CYP1A1 and CYP2B) induced by some POPs (DDTs and PCBs) and emerging contaminants (PBDEs) in fibroblast cell cultures of striped dolphin (Stenella coeruleoalba) and bottlenose dolphin (Tursiops truncatus). Immunofluorescence was quantified with a specially designed Olympus macro, DetectIntZ. A major result was the possibility of using this "in vitro" assay to quantify induction of endogenous proteins.

  4. [Designing and clinical testing of immune-enzyme and immunofluorescence test systems for serodiagnosis of ixodes borreliosis].

    Science.gov (United States)

    Pomelova, V G; Kharitonenkov, I G; Sadykbekova, R K; Bychenkova, T A; Anan'eva, L P; Sokolova, M V; Osin, N S

    2004-01-01

    The methods of immune enzyme assay (MIEA) and of lanthanide immunofluorescence analysis (LIFA) were used to work out the test systems for the detection (in blood serum of patients) of specific IgM IgG antibodies to the B. burgdorferi spirochete--a causative agent of ixodic borrelioses. The test system was clinically tested versus the indirect immunofluorescence reaction (IIFR) and commercial immune enzyme test system (CIET). The results of antibodies' detection were shown to correlate with the analysis data for the same sera in IIFR and to be in line with a real presence or absence of the disease. Test systems based on LIFA were proven to be most sensitive and specific.

  5. A new European plant-specific emission inventory of biogenic volatile organic compounds for use in atmospheric transport models

    Directory of Open Access Journals (Sweden)

    M. Karl

    2009-06-01

    Full Text Available We present a new European plant-specific emission inventory for isoprene, monoterpenes, sesquiterpenes and oxygenated VOC (OVOC, on a spatial resolution of 0.089×0.089 degrees, for implementation in atmospheric transport models. The inventory incorporates more accurate data on foliar biomass densities from several litterfall databases that became available in the last years for the main tree species in Europe. A bioclimatic correction factor was introduced to correct the foliar biomass densities of trees and crops for the different plant growth conditions that can be found in Pan-Europe. Long-term seasonal variability of agriculture and forest emissions was taken into account by implementing a new growing season concept. The 2004–2005 averaged annual total biogenic volatile organic compound (BVOC emissions for the Pan-European domain are estimated to be about 12 Tg with a large contribution from the OVOC class of about 4.5 Tg and from monoterpenes of about 4 Tg. Annual isoprene emissions are found to be about 3.5 Tg, insensitive to the chosen emission algorithm. Emissions of OVOC were found to originate to a large extent from agriculture. Further experiments on crop emissions should be carried out to check the validity of the applied standard emission factors. The new inventory aims at a fully transparent and verifiable aggregation of detailed land use information and at the inclusion of plant-specific emission data. Though plant-specific land use data is available with relatively high accuracy, a lack of experimental biomass densities and emission data on terpenes, sesquiterpenes and oxygenated VOC, in particular for agricultural plants, currently limits the setup of a highly accurate plant-specific emission inventory.

  6. 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

  7. Degradation of sulfamethoxazole using ozone and chlorine dioxide - Compound-specific stable isotope analysis, transformation product analysis and mechanistic aspects.

    Science.gov (United States)

    Willach, Sarah; Lutze, Holger V; Eckey, Kevin; Löppenberg, Katja; Lüling, Michelle; Terhalle, Jens; Wolbert, Jens-Benjamin; Jochmann, Maik A; Karst, Uwe; Schmidt, Torsten C

    2017-10-01

    The sulfonamide antibiotic sulfamethoxazole (SMX) is a widely detected micropollutant in surface and groundwaters. Oxidative treatment with e.g. ozone or chlorine dioxide is regularly applied for disinfection purposes at the same time exhibiting a high potential for removal of micropollutants. Especially for nitrogen containing compounds such as SMX, the related reaction mechanisms are largely unknown. In this study, we systematically investigated reaction stoichiometry, product formation and reaction mechanisms in reactions of SMX with ozone and chlorine dioxide. To this end, the neutral and anionic SMX species, which may occur at typical pH-values of water treatment were studied. Two moles of chlorine dioxide and approximately three moles of ozone were consumed per mole SMX degraded. Oxidation of SMX with ozone and chlorine dioxide leads in both cases to six major transformation products (TPs) as revealed by high-resolution mass spectrometry (HRMS). Tentatively formulated TP structures from other studies could partly be confirmed by compound-specific stable isotope analysis (CSIA). However, for one TP, a hydroxylated SMX, it was not possible by HRMS alone to identify whether hydroxylation occurred at the aromatic ring, as suggested in literature before, or at the anilinic nitrogen. By means of CSIA and an analytical standard it was possible to identify sulfamethoxazole hydroxylamine unequivocally as one of the TPs of the reaction of SMX with ozone as well as with chlorine dioxide. H-abstraction and electron transfer at the anilinic nitrogen are suggested as likely initial reactions of ozone and chlorine dioxide, respectively, leading to its formation. Oxidation of anionic SMX with ozone did not show any significant isotopic fractionation whereas the other reactions studied resulted in a significant carbon isotope fractionation. Copyright © 2017 Elsevier Ltd. All rights reserved.

  8. Validation of adsorbents for sample preconcentration in compound-specific isotope analysis of common vapor intrusion pollutants.

    Science.gov (United States)

    Klisch, Monika; Kuder, Tomasz; Philp, R Paul; McHugh, Thomas E

    2012-12-28

    Isotope ratios of volatile organic compounds (VOCs) in the environment are often of interest in contaminant fate studies. Adsorbent preconcentration-thermal desorption of VOCs can be used to collect environmental vapor samples for compound-specific isotope analysis (CSIA). While active adsorbent samplers offer logistic benefits in handling large volumes of air, their performance in preserving VOCs isotope ratios was not previously tested under sampling conditions corresponding to typical indoor air sampling conditions. In this study, the performance of selected adsorbents was tested for preconcentration of TCE (for determination of C and Cl isotope ratios), PCE (C and Cl) and benzene (C and H). The key objective of the study was to identify the adsorbent(s) permitting preconcentration of the target VOCs present in air at low μg/m(3) concentrations, without significant alteration of their isotope ratios. Carboxen 1016 was found to perform well for the full range of tested parameters. Carboxen 1016 can be recommended for sampling of TCE, PCE and benzene, for CSIA, from air volumes up to 100 L. Variable extent of isotope ratio alteration was observed in the preconcentration of the target VOCs on Carbopack B and Carbopack X, resulting from partial analyte loss via adsorbent bed breakthrough and (possibly) via incomplete desorption. The results from testing the Carbopack B and Carbopack X highlight the need of adsorbent performance validation at conditions fully representative of actual sample collection conditions, and caution against extrapolation of performance data toward more challenging sampling conditions. Copyright © 2012 Elsevier B.V. All rights reserved.

  9. Image based quantitative reader for Lateral flow immunofluorescence assay.

    Science.gov (United States)

    Chowdhury, Kaushik Basak; Joseph, Jayaraj; Sivaprakasam, Mohanasankar

    2015-08-01

    Fluorescence Lateral flow immunoassays (LFIA) have wide range of applications in point-of-care testing (POCT). An integrated, motion-free, accurate, reliable reader that performs automated quantitative analysis of LFIA is essential for POCT diagnosis. We demonstrate an image based quantitative method to read the lateral flow immunofluorescence test strips. The developed reader uses line laser diode module to illuminate the LFIA test strip having fluorescent dye. Fluorescence light coming from the region of interest (ROI) of the LFIA test strip was filtered using an emission filter and imaged using a camera following which images were processed in computer. A dedicated control program was developed that automated the entire process including illumination of the test strip using laser diode, capturing the ROI of the test strip, processing and analyzing the images and displaying of results. Reproducibility of the reader has been evaluated using few reference cartridges and HbA1c (Glycated haemoglobin) test cartridges. The proposed system can be upgraded to a compact reader for widespread testing of LFIA test strips.

  10. [Analysis of direct immunofluorescence tests for trachoma diagnosis].

    Science.gov (United States)

    Medina, N H; Gentil, R M; Caraça, M; Suzuki, C K; Melles, H H

    1996-04-01

    For the confirmation of trachoma foci in places where no previous cases had been reported, the State Health Secretariat of S. Paulo makes provision for the realization of laboratory exams, particularly because the disease was considered to have been eradicated from the State in the seventies. During the epidemiological investigations, conjunctival scrapings were collected from the subjects with inflammatory trachoma (TF/TI), clinically diagnosed. The results of the immunofluorescence (DFA) exams were analysed in the light of the frequency of the appropriate exams and their positive results, by the quantity of elementary bodies (EB) found. A total of 385 slides were studied, the criteria for positivity being 5 or more EBs. The test's sensitivity was 19.9%. The DFA test is considered to be the best laboratorial exam to be used in field work, though it does not show a sufficient sensitivity to confirm all clinically diagnosed cases of trachoma; it can only confirm the circulation of the aetiological agent within a community. In endemic areas the clinical diagnosis continues to be the criterion for case confirmation.

  11. Unequivocal identification of the active and inactive centromere of a dicentric chromosome by simultaneous CENP-C immunofluorescence and FISH

    Energy Technology Data Exchange (ETDEWEB)

    Page, S.L.; Shaffer, L.G. [Baylor College of Medicine, Houston, TX (United States); Earnshaw, W.C. [Johns Hopkins Univ. School of Medicine, Baltimore, MD (United States)] [and others

    1994-09-01

    There have been many reports of stable dicentric chromosomes resulting from translocations in humans. The stability of these chromosomes has been attributed to the inactivation of one centromere, yielding a functionally monocentric chromosome. We ascertained a patient with the karyotype 45,X/45,X,der(X)t(X;15)(Xpter{yields}Xq26::15p11{yields}15qter). The derivative chromosome was apparently dicentric by G-band analysis. A primary constriction at the X centromere and lack of a constriction corresponding to the 15 centromere suggested that the 15 centromere was inactive, while the X centromere remained active. Centromere Protein C(CENP-C) is normally present at all human centromeres and is localized to the inner kinetochore plate. Previous immunofluorescence studies of a stable isodicentric chromosome showed CENP-C to be present at the active centromere, while no staining was observed at the inactive centromere. By combining immunofluorescence detection of CENP-C and multi-color fluorescence in situ hybridization (FISH) with chromosome-specific {alpha} satellite DNA probes, we have unequivocally identified the active centromere in the (X;15) translocation. Metaphases were prepared by cytocentrifugation of hypotonically swelled lymphoblasts. Indirect immunofluorescence, performed using polyclonal rabbit antiserum raised against CENP-C, and FISH with {alpha}-satellite probes specific for the centromeres of chromosomes 15 (pTRA-20,pTRA-25) and X (DXZ1) revealed CENP-C immunofluorescence signals at the X {alpha}-satellite probe, with no CENP-C detected at the 15 centromere on the translocation in this cell line. This study is the first to demonstrate the specificity of CENP-C to the active centromere in a nonhomologous dicentric rearrangement and further establishes CENP-C as an essential component of a functional human centromere.

  12. Understanding the complexity of antigen retrieval of DNA methylation for immunofluorescence-based measurement and an approach to challenge.

    Science.gov (United States)

    Çelik, Selcen

    2015-01-01

    Cytosine methylation (5-methylcytosine, 5meC) in the CpG-rich regions of the mammalian genome is an important epigenetic mechanism playing roles in transcription regulation and genomic stability. The abnormalities in DNA methylation can occur in various types of cancer and some genetic diseases. The measurement of DNA methylation is therefore important and there is a range of methodologies used to detect DNA methylation. Many methods based on bisulfite treatment appeared with a lack of specificity after recent discoveries of various modifications of methylated cytosine, however there are new treatments developed to overcome this limitation. Immunofluorescence is currently known to be able to specifically detect DNA methylation as it uses different antibodies against 5meC and its derivatives, but it is a semi-quantitative method. Immunofluorescence protocols commonly include fixation of cells followed by permeabilisation, antigen retrieval, and treatments with antibodies. Establishing the strategy for antigen retrieval of immunofluorescence is important to unmask epitopes (i.e. 5meC) from other proteins, and therefore to access the antigen of interest. There are many approaches used for antigen retrieval induced by acid, enzyme and/or heat. The selection of antigen retrieval method can depend on a variety of such antigen-based or cell-based conditions, since the dynamic structure of DNA and chromatin accounts for the complexity of involved proteins to mask the epitope. This review aims to specifically focus on the complexity of in situ detection of DNA methylation by immunofluorescence-based methods using antigen retrieval with the current understanding of DNA methylation mechanism, and suggests conditions for antigenic retrieval of 5meC epitope. Copyright © 2014 Elsevier B.V. All rights reserved.

  13. Cell Cycle Dynamics of Proteins and Post-translational Modifications Using Quantitative Immunofluorescence.

    Science.gov (United States)

    Akopyan, Karen; Lindqvist, Arne; Müllers, Erik

    2016-01-01

    Immunofluorescence can be a powerful tool to detect protein levels, intracellular localization, and post-translational modifications. However, standard immunofluorescence provides only a still picture and thus lacks temporal information. Here, we describe a method to extract temporal information from immunofluorescence images of fixed cells. In addition, we provide an optional protocol that uses micropatterns, which increases the accuracy of the method. These methods allow assessing how protein levels, intracellular localization, and post-translational modifications change through the cell cycle.

  14. Correlation of bulk sedimentary and compound-specific δ13C values indicates minimal pre-aging of n-alkanes in a small tropical watershed

    Science.gov (United States)

    Lane, Chad S.; Horn, Sally P.; Taylor, Zachary P.; Kerr, Matthew T.

    2016-08-01

    Inherent to sedimentary compound-specific isotopic proxies of paleoecological and paleoclimate change is the assumption that biomarkers are coeval with the surrounding sedimentary matrix. Compound-specific radiocarbon analyses of lake and nearshore marine sediments have indicated a potential offset between the ages of terrestrial biomarkers and their surrounding sediments that could confound efforts to establish strong chronological control for compound-specific isotopic data. We conducted high-resolution compound-specific δ13C analyses of n-alkanes (δ13Calkane) in a well-studied sediment core from Laguna Castilla, Dominican Republic, and compared the results to bulk sedimentary δ13C (δ13Cbulk), fossil pollen, and sediment geochemistry to assess potential 'pre-aging' of alkanes in the terrestrial environment prior to deposition. We found significant positive correlations between δ13Cbulk values and δ13Calkane values, indicating little or no temporal lag in the response of δ13Calkane values to vegetation change and thus little or no offset in the age of terrestrially-derived compounds and the organic fraction of the sedimentary matrix. While this study highlights the need for further research into the variables affecting age offsets between proxy data, we propose the comparison of δ13Cbulk and δ13Calkane values as a method to assess potential age offsets between compound-specific and other proxy datasets, particularly in small watersheds with sediment records containing a high proportion of allochthonous organic matter. This method is more available to researchers than obtaining numerous compound-specific radiocarbon analyses, which are costly and not a routine service offered by radiocarbon facilities.

  15. Microtubules in Plant Cells: Strategies and Methods for Immunofluorescence, Transmission Electron Microscopy, and Live Cell Imaging.

    Science.gov (United States)

    Celler, Katherine; Fujita, Miki; Kawamura, Eiko; Ambrose, Chris; Herburger, Klaus; Holzinger, Andreas; Wasteneys, Geoffrey O

    2016-01-01

    Microtubules (MTs) are required throughout plant development for a wide variety of processes, and different strategies have evolved to visualize and analyze them. This chapter provides specific methods that can be used to analyze microtubule organization and dynamic properties in plant systems and summarizes the advantages and limitations for each technique. We outline basic methods for preparing samples for immunofluorescence labeling, including an enzyme-based permeabilization method, and a freeze-shattering method, which generates microfractures in the cell wall to provide antibodies access to cells in cuticle-laden aerial organs such as leaves. We discuss current options for live cell imaging of MTs with fluorescently tagged proteins (FPs), and provide chemical fixation, high-pressure freezing/freeze substitution, and post-fixation staining protocols for preserving MTs for transmission electron microscopy and tomography.

  16. [Comparative value of ELISA technique and immunofluorescence for the detection in the foci of human African trypanosomiasis].

    Science.gov (United States)

    Mangenot, M; Chaize, J; Desfontaine, M; Duvallet, G; Moreau, J P

    1979-01-01

    As the parasitological diagnosis method consisting in the detection of the parasite in people suffering from sleeping sickness did not prove entirely reliable, the authors have tested a new immunoenzymatic diagnosis method called ELISA. 41 sera from clinical suspects have been tested (parasite having been found in 16 of them). Positive and negative levels have been tested by comparison with indirect immunofluorescence. The following results, given in optical density, have been obtained: negative cases less than 36 less than or equal to uncertain cases less than or equal to 40 less than positive cases. When using these date, correlation with immunofluorescence proves to be excellent with a sensibility between 95 and 97,5 p. 100 and a specificity between 97,5 and 99 p. 100.

  17. Compound-specific isotopes of fatty acids as indicators of trophic interactions in the East China Sea ecosystem

    Science.gov (United States)

    Wu, Ying; Wang, Na; Zhang, Jing; Wan, Ruijing; Dai, Fangqun; Jin, Xianshi

    2016-09-01

    The composition and compound-specific isotopes of fatty acids were studied within food webs in the East China Sea. Lipid-normalized stable carbon isotopes of total organic carbon had a good correlation with trophic level. Variations in fatty acid compositions among diff erent species were observed but were unclear. Diff erent dietary structures could be traced from molecular isotopes of selected fatty acids in the Shiba shrimp ( Matapenaeus joyneri), the coastal mud shrimp ( Solenocera crassicornis) and the northern Maoxia shrimp ( Acetes chinensis). Both M. joyneri and S. crassicornis are mainly benthos feeders, while A. chinensis is a pelagic species, although they have a similar fatty acid composition. There was a good correlation for isotopes of arachidonic acid (C20:4n6; ARA) and docosahexaenoic acid (C22:6n3; DHA) among pelagic species from higher trophic levels. The isotopic compositions of DHA in benthic species were more negative than those of pelagic species at the same trophic level. The fact that the diet of benthic species contains more degraded items, the carbon isotopes of which are derived from a large biochemical fraction, may be the reason for this variation. A comparative study of benthic and pelagic species demonstrated the diff erent carbon sources in potential food items and the presence of a more complex system at the water-sediment interface.

  18. Rhizoctonia solani and Bacterial Inoculants Stimulate Root Exudation of Antifungal Compounds in Lettuce in a Soil-Type Specific Manner

    Directory of Open Access Journals (Sweden)

    Saskia Windisch

    2017-06-01

    Full Text Available Previous studies conducted on a unique field site comprising three contrasting soils (diluvial sand DS, alluvial loam AL, loess loam LL under identical cropping history, demonstrated soil type-dependent differences in biocontrol efficiency against Rhizoctonia solani-induced bottom rot disease in lettuce by two bacterial inoculants (Pseudomonas jessenii RU47 and Serratia plymuthica 3Re-4-18. Disease severity declined in the order DS > AL > LL. These differences were confirmed under controlled conditions, using the same soils in minirhizotron experiments. Gas chromatography-mass spectrometry (GC-MS profiling of rhizosphere soil solutions revealed benzoic and lauric acids as antifungal compounds; previously identified in root exudates of lettuce. Pathogen inoculation and pre-inoculation with bacterial inoculants significantly increased the release of antifungal root exudates in a soil type-specific manner; with the highest absolute levels detected on the least-affected LL soil. Soil type-dependent differences were also recorded for the biocontrol effects of the two bacterial inoculants; showing the highest efficiency after double-inoculation on the AL soil. However, this was associated with a reduction of shoot growth and root hair development and a limited micronutrient status of the host plants. Obviously, disease severity and the expression of biocontrol effects are influenced by soil properties with potential impact on reproducibility of practical applications.

  19. Evaluation of the performance of high temperature conversion reactors for compound-specific oxygen stable isotope analysis.

    Science.gov (United States)

    Hitzfeld, Kristina L; Gehre, Matthias; Richnow, Hans-Hermann

    2017-05-01

    In this study conversion conditions for oxygen gas chromatography high temperature conversion (HTC) isotope ratio mass spectrometry (IRMS) are characterised using qualitative mass spectrometry (IonTrap). It is shown that physical and chemical properties of a given reactor design impact HTC and thus the ability to accurately measure oxygen isotope ratios. Commercially available and custom-built tube-in-tube reactors were used to elucidate (i) by-product formation (carbon dioxide, water, small organic molecules), (ii) 2nd sources of oxygen (leakage, metal oxides, ceramic material), and (iii) required reactor conditions (conditioning, reduction, stability). The suitability of the available HTC approach for compound-specific isotope analysis of oxygen in volatile organic molecules like methyl tert-butyl ether is assessed. Main problems impeding accurate analysis are non-quantitative HTC and significant carbon dioxide by-product formation. An evaluation strategy combining mass spectrometric analysis of HTC products and IRMS (18)O/(16)O monitoring for future method development is proposed.

  20. The Response of Creatine Kinase Specific Activity in Rat Pituitary to Estrogenic Compounds and Vitamin D Less-Calcemic Analogs

    Directory of Open Access Journals (Sweden)

    D. Somjen

    2009-01-01

    Full Text Available We examined the response of rat female pituitary at different metabolic stages to treatments with estrogenic compounds and vitamin D analogs. Immature or ovariectomized (Ovx female rats responded by increased creatine kinase specific activity (CK to estradiol-17β (E2, genistein (G, daidzein (D, biochainin A (BA, quecertin (Qu, carboxy- G (cG, carboxy- BA (cBA, and raloxifene (Ral. The response was inhibited when Ral was injected together with the estrogens. CK was increased when hormones were injected daily into Ovx rats for 4 different time periods. Pretreatment with the less-calcemic vitamin D analogs JK 1624 F2-2 (JKF or QW 1624 F2-2 (QW followed by estrogenic injection resulted in increased response and sensitivity to E2 and loss of inhibition of E2 by Ral. CK was also increased by feeding with E2 or licorice or its components dose- and time- dependent in immature or Ovxrats. Diabetic female rats did not respond to increased doses of E2. In conclusion, rat female pituitary is estrogens-responsive organ, suggesting to consider its response for HRT in postmenopausal women for both beneficial and hazardous aspects.

  1. Compound-specific amino acid isotopic analyses of invertebrates in the Chukchi Sea: New insights on food web dynamics

    Science.gov (United States)

    Zhang, M.; Cooper, L. W.; Biasatti, D. M.; Kedra, M.; Grebmeier, J. M.

    2016-02-01

    Food web dynamics in the Chukchi Sea have been previously evaluated using bulk analysis of stable carbon and nitrogen isotopes of organisms. However, recent advances in compound-specific stable isotope analysis of amino acids indicate the potential to better identify the contributions of different dietary sources (e.g., pelagic vs. benthic, ice algae vs. phytoplankton) and to resolve complexities of food web structure that are difficult to address with bulk isotope analysis. Here we combine amino acid δ13C and δ15N data measured from primary producers and tissues of bivalves, polychaetes and other benthic invertebrates collected during two cruises in the summer of 2013 and 2015 in the Pacific Arctic. The results showed spatial variation of carbon isotope values in amino acids with difference up to 6 per mil for each individual species or taxa studied, indicating a shift in the food-web baseline geographically. Furthermore, the spatial variation in isotopic values was related to environmental factors, specifically sea ice extent, and total organic carbon, total organic nitrogen and the carbon/nitrogen ratio of the organic fractions of surface sediments. Results also indicated that trophic levels, as estimated by differences in the nitrogen isotope composition of glutamic acid and phenylalanine [Δ15Nglu-phe (δ15Nglu - δ15Nphe)], varied spatially by 0.5 to 1.5 trophic levels for certain species or taxa such as Macoma calcarea, Maldanidae and Ampelisca, indicating trophic level shifts that were associated with the food quality of organic matter in the organic fraction of the sediments. These results can be potentially used to predict future food web change in this high latitude marine system that is known for its ecological importance and on-going environmental changes, including warming and sea ice decline.

  2. The specific influence of carbon dioxide and carbamate compounds on the buffer power and Bohr effects in human haemoglobin solutions

    Science.gov (United States)

    Rossi-Bernardi, L.; Roughton, F. J. W.

    1967-01-01

    1. The difference of pH (ΔpH) between human deoxygenated haemoglobin (Hb) and oxygenated haemoglobin (O2Hb) solutions when equilibrated with physiological pressures of carbon dioxide is (experimentally) much less than previously supposed. 2. This smaller ΔpH is in contradiction to Wyman's (1948) theoretical calculations, wherein no allowance was made for the specific effect of carbamino-compounds on the amount of base neutralized by haemoglobin. Other previous authorities have also neglected this factor, which when properly allowed for restores the role of carbamino-compounds in CO2 transport practically to that previously estimated by Ferguson & Roughton (1934a, b). 3. At a given pH and PCO2, more CO2 is bound by Hb solutions than by O2Hb, the difference increasing with pH. This result provides further, and seemingly decisive, evidence that the bound-CO2 in the blood other than HCO3— (i.e. x-bound CO2) is oxygen-linked. 4. According to a modified form of the Henderson—Hasselbalch equation for haemoglobin solutions [Formula: see text]. The value of pK1′ is the same in Hb as in O2Hb solution and from the data in 3 is found to have the value 6·15 at 37° C. 5. The difference between the titration curves of O2Hb and Hb (ΔX̄), at a given pH had been hitherto supposed to be the same in presence of CO2 as in its absence. Our experiments show, however, that ΔX̄ is less in presence of CO2 and at pH > 7·5 may even change sign. This paradoxical effect is also explicable, at any rate semi-quantitatively, by the effect of carbamino compounds on the buffer power, according to the theory put forward in the paper. 6. The results show that the buffer power (dB/dpH) of haemoglobin solution under physiological conditions is 20-30% greater than previously estimated, and this also is in line with the new theory. 7. In graphs of total CO2 versus PCO2 in haemoglobin solutions (or blood) it has been customary to suppose that points on straight lines radiating from the

  3. Monitoring biodegradation of ethene and bioremediation of chlorinated ethenes at a contaminated site using compound-specific isotope analysis (CSIA)

    Science.gov (United States)

    Mundle, S.O.C.; Johnson, T.; Lacrampe-Couloume, G.; Perez-De-Mora, A.; Duhamel, M.; Edwards, E.A.; McMaster, M.L.; Cox, E.; Revesz, K.; Lollar, B. Sherwood

    2012-01-01

    Chlorinated ethenes are commonly found in contaminated groundwater. Remediation strategies focus on transformation processes that will ultimately lead to nontoxic products. A major concern with these strategies is the possibility of incomplete dechlorination and accumulation of toxic daughter products (cis-1,2-dichloroethene (cDCE), vinyl chloride (VC)). Ethene mass balance can be used as a direct indicator to assess the effectiveness of dechlorination. However, the microbial processes that affect ethene are not well characterized and poor mass balance may reflect biotransformation of ethene rather than incomplete dechlorination. Microbial degradation of ethene is commonly observed in aerobic systems but fewer cases have been reported in anaerobic systems. Limited information is available on the isotope enrichment factors associated with these processes. Using compound-specific isotope analysis (CSIA) we determined the enrichment factors associated with microbial degradation of ethene in anaerobic microcosms (ε = −6.7‰ ± 0.4‰, and −4.0‰ ± 0.8‰) from cultures collected from the Twin Lakes wetland area at the Savannah River site in Georgia (United States), and in aerobic microcosms (ε = −3.0‰ ± 0.3‰) from Mycobacterium sp. strain JS60. Under anaerobic and aerobic conditions, CSIA can be used to determine whether biotransformation of ethene is occurring in addition to biodegradation of the chlorinated ethenes. Using δ13C values determined for ethene and for chlorinated ethenes at a contaminated field site undergoing bioremediation, this study demonstrates how CSIA of ethene can be used to reduce uncertainty and risk at a site by distinguishing between actual mass balance deficits during reductive dechlorination and apparent lack of mass balance that is related to biotransformation of ethene.

  4. Compound-Specific Isotope Analysis (CSIA Application for Source Apportionment and Natural Attenuation Assessment of Chlorinated Benzenes

    Directory of Open Access Journals (Sweden)

    Luca Alberti

    2017-11-01

    Full Text Available In light of the complex management of chlorobenzene (CB contaminated sites, at which a hydraulic barrier (HB for plumes containment is emplaced, compound-specific stable isotope analysis (CSIA has been applied for source apportionment, for investigating the relation between the upgradient and downgradient of the HB, and to target potential CB biodegradation processes. The isotope signature of all the components potentially involved in the degradation processes has been expressed using the concentration-weighted average δ13C of CBs + benzene (δ13Csum. Upgradient of the HB, the average δ13Csum of −25.6‰ and −29.4‰ were measured for plumes within the eastern and western sectors, respectively. Similar values were observed for the potential sources, with δ13Csum values of −26.5‰ for contaminated soils and −29.8‰ for the processing water pipeline in the eastern and western sectors, respectively, allowing for apportioning of these potential sources to the respective contaminant plumes. For the downgradient of the HB, similar CB concentrations but enriched δ13Csum values between −24.5‰ and −25.9‰ were measured. Moreover, contaminated soils showed a similar δ13Csum signature of −24.5‰, thus suggesting that the plumes likely originate from past activities located in the downgradient of the HB. Within the industrial property, significant δ13C enrichments were measured for 1,2,4-trichlorobenzene (TCB, 1,2-dichlorobenzene (DCB, 1,3-DCB, and 1,4-DCBs, thus suggesting an important role for anaerobic biodegradation. Further degradation of monochlorobenzene (MCB and benzene was also demonstrated. CSIA was confirmed to be an effective approach for site characterization, revealing the proper functioning of the HB and demonstrating the important role of natural attenuation processes in reducing the contamination upgradient of the HB.

  5. Specific skeletal muscle sphingolipid compounds in energy expenditure regulation and weight gain in Native Americans of Southwestern heritage.

    Science.gov (United States)

    Heinitz, S; Piaggi, P; Vinales, K L; Basolo, A; Spranger, J; Piomelli, D; Krakoff, J; Jumpertz von Schwartzenberg, R

    2017-10-01

    In animal models, a role in the regulation of energy expenditure (EE) has been ascribed to sphingolipids, active components of cell membranes participating in cellular signaling. In humans, it is unknown whether sphingolipids have a role in the modulation of EE and, consequently, influence weight gain. The present study investigated the putative association of EE and weight gain with sphingolipid levels in the human skeletal muscle, a component of fat-free mass (the strongest determinant of EE), in adipose tissue and plasma. Twenty-four-hour EE, sleeping metabolic rate (SMR) and resting metabolic rate (RMR) were assessed in 35 healthy Native Americans of Southwestern heritage (24 male; 30.2±7.73 years). Sphingolipid (ceramide, C; sphingomyelin, SM) concentrations were measured in skeletal muscle tissue, subcutaneous adipose tissue and plasma samples. After 6.68 years (0.26-12.4 years), follow-up weights were determined in 16 participants (4 females). Concentrations of C24:0, SM18:1/26:1 and SM18:0/24:1 in muscle were associated with 24-h EE (r=-0.47, P=0.01), SMR (r=-0.59, P=0.0008) and RMR (r=-0.44, P=0.01), respectively. Certain muscle sphingomyelins also predicted weight gain (for example, SM18:1/23:1, r=0.74, P=0.004). For specific muscle sphingomyelins that correlated with weight gain and EE (SM18:1/23:0, SM18:1/23:1 and SMR, r=-0.51, r=-0.41, respectively, all Pmuscle and adipose tissue sphingolipid compounds are associated with EE and weight gain in Native Americans of Southwestern heritage. Further studies are warranted to investigate whether sphingolipids of different body compartments act in concert to modulate energy balance in humans.

  6. Autoradiography and immunofluorescence combined for autecological study of single cell activity with Nitrobacter as a model system.

    Science.gov (United States)

    Fliermans, C B; Schmidt, E L

    1975-10-01

    Specific detection of a particular bacterium by immunofluorescence was combined with estimation of its metabolic activity by autoradiography. The nitrifying bacteria Nitrobacter agilis and N. winogradskyi were used as a model system. Nitrobacter were incubated with NaH14CO3 and 14CO2 prior to study. The same preparations made for autoradiograms were stained with fluorescent antibodies specific for the Nitrobacter species. Examination by epifluorescence and transmitted dark-field microscopy revealed Nitrobacter cells with and without associated silver grains. Direct detection and simultaneous evaluation of metabolic activity of Nitrobacter was demonstrated in pure cultures, in a simple mixed culture, and in a natural soil.

  7. Autoradiography and Immunofluorescence Combined for Autecological Study of Single Cell Activity with Nitrobacter as a Model System1

    Science.gov (United States)

    Fliermans, C. B.; Schmidt, E. L.

    1975-01-01

    Specific detection of a particular bacterium by immunofluorescence was combined with estimation of its metabolic activity by autoradiography. The nitrifying bacteria Nitrobacter agilis and N. winogradskyi were used as a model system. Nitrobacter were incubated with NaH14CO3 and 14CO2 prior to study. The same preparations made for autoradiograms were stained with fluorescent antibodies specific for the Nitrobacter species. Examination by epifluorescence and transmitted dark-field microscopy revealed Nitrobacter cells with and without associated silver grains. Direct detection and simultaneous evaluation of metabolic activity of Nitrobacter was demonstrated in pure cultures, in a simple mixed culture, and in a natural soil. Images PMID:1103733

  8. Acquisition of species-specific perfume blends: influence of habitat-dependent compound availability on odour choices of male orchid bees (Euglossa spp.).

    Science.gov (United States)

    Pokorny, T; Hannibal, M; Quezada-Euan, J J G; Hedenström, E; Sjöberg, N; Bång, J; Eltz, T

    2013-06-01

    Male orchid bees (Euglossini, Apidae, Hymenoptera) expose species-specific blends of volatile chemicals (perfume bouquets) during their courtship display. The perfumes are acquired by collecting fragrant substances from environmental sources, which are then accumulated in specialised hind leg pouches. To balance the perfume composition, the males need to find and collect the required substances in specific relative amounts while facing seasonal, local or habitat-dependent differences in compound availability. Experience-dependent choice of odours, i.e. 'learned avoidance' of recently collected components, has been proposed as the mechanism that mediates the accumulation of the stereotypical compound ratios. In the present study, we used the presence of certain compounds in male hind leg pouches as proxy for the respective local compound availability, and investigated whether differences in content are correlated with differences in chemical choice assays. Our results suggest that volatile availability differs between localities (n = 16) as well as habitats (n = 2; coastal vs. inland) across the Yucatán peninsula, Mexico, for both studied species. Male Euglossa dilemma showed a pronounced preference for benzyl benzoate and eugenol at locations where those compounds were rare in hind leg extracts, as predicted by the learned avoidance model. No equivalent correlations were found for Euglossa viridissima. This is the first study to combine chemical analyses of perfumes with bioassays of odour choice. It strengthens the view that negative feedback from collected odours modifies future chemical choice and helps males to acquire specific perfume blends.

  9. Enhancement of tight junctional barrier function by micronutrients: compound-specific effects on permeability and claudin composition.

    Directory of Open Access Journals (Sweden)

    Joanna Mercado

    Full Text Available Amid an increasing number of reports in the literature concerning epithelial barrier enhancement by various nutrient compounds, there has never been a study performing side-by-side comparisons of these agents in a single epithelial model. We compare five nutrient compounds (previously reported in various epithelial models to enhance barrier function regarding their ability to increase transepithelial electrical resistance (R(t and decrease transepithelial mannitol permeability (J(m across LLC-PK₁ renal epithelial cell layers. The effects of these nutrients on the abundance of various tight junctional proteins are also compared. In the overall group of nutrients tested--zinc, indole, quercetin, butyrate and nicotine--only nicotine failed to improve barrier function by either parameter. Nicotine also was without effect on tight junctional proteins. Quercetin simultaneously increased R(t and decreased J(m. Zinc, butyrate and indole only exhibited statistically significant enhancement of R(t. Each of these four effective nutrient compounds had unique patterns of effects on the panel of tight junctional proteins studied. No two compounds produced the same pattern of effects. This unique pattern of effects on tight junctional complex composition by each compound establishes the chance for additive or even synergistic improvement of barrier function by combinations of compounds. A synergistic effect of the combination of quercetin and zinc on R(t is shown.

  10. A comparison between immunofluorescence staining on smears from Membrana nictitans (M3 test), immunohistopathology and routine pathology in cats with suspected feline infectious peritonitis (FIP).

    Science.gov (United States)

    Hök, K

    1991-01-01

    An indirect immunofluorescence method using smears from membrana nictitans (M3 test) to diagnose feline corona virus (FCV) infection was compared with immunohistopathology (using indirect immunofluorescence assay (IFFA) performed on organs (IFO], and routine pathology (RP) in cats with suspected feline infectious peritonitis (FIP). A close correlation between the 2 immunofluorescence methods (IFO and M3) was observed. Although the M3 test requires samples from only 1 organ per animal, both the sensitivity and specificity were high (80%), when compared to IFO (using samples from an average of 5 organs per animal). In 21% of the cats with suspected FIP typical pathological lesions were found. As the M3 test is relatively easy to perform, it could reduce work-load of pathology laboratories and provide valuable data for clinical and epidemiological use.

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

    Science.gov (United States)

    Sebastian, Wendy; Roy, Atanu; Kini, Usha; Mullick, Shalini

    2010-01-01

    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. 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. 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. 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. The antinuclear antibody indirect immunofluorescence (ANA - IIF) patterns obtained were projectable to visualize a certain spectrum of specific antibodies such as

  12. Comparison of an enzyme-linked immunosorbent assay, an immunofluorescence assay and a hemagglutination inhibition assay for detection of antibodies to K-papovavirus in mice.

    NARCIS (Netherlands)

    J. Groen (Jan); A.D.M.E. Osterhaus (Albert); H.W.J. Broeders; H.E.M. Spijkers (Ine)

    1989-01-01

    textabstractThe sensitivity of a newly developed enzyme-linked immunosorbent assay (ELISA) for detection of antibody to K virus was compared with the sensitivities of an immunofluorescence assay (IFA) and a hemagglutination inhibition assay (HIA). Specific pathogen-free BALB/c RIVM mice, 5 weeks

  13. 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

    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...

  14. Functional connectivity of coral reef fishes in a tropical seascape assessed by compound-specific stable isotope analyses

    KAUST Repository

    McMahon, Kelton W.

    2011-01-01

    . ehrenbergii populations in the Red Sea. The compound-specific SIA approach presented in this thesis will be particularly valuable for tracking the movement of species and life-stages not amenable to conventional tagging techniques. This thesis provides quantitative scientific support for establishing realistic population connectivity models that can be used to design effective marine reserve networks.

  15. Site specific X-ray induced changes in organic and metal organic compounds and their influence on global radiation damage

    Energy Technology Data Exchange (ETDEWEB)

    Heintz, Desiree Ellen

    2012-07-15

    The aim of this work was to systematically investigate the effects of specific and global X-ray radiation damage to biological samples and obtain a conclusive model to describe the underlying principles. Based on the systematic studies performed in this work, it was possible to propose two conclusive mechanisms to describe X-ray induced photoreduction and global radiation damage. The influence of chemical composition, temperature and solvent on X-ray induced photoreduction was investigated by X-ray Absorption Near Edge Spectroscopy and single crystal X-ray diffraction of two B12 cofactors - cyano- and methylcobalamin - as well as iron(II) and iron(III) complexes. The obtained results revealed that X-ray induced photoreduction is a ligand dependent process, with a redox reaction taking place within the complex. It could further be shown that selective hydrogen abstraction plays an important role in the process of X-ray induced photoreduction. Based on the experimental results of this work, a model to describe X-ray induced photoreduction of metal organic complexes could be proposed. The process of X-ray induced hydrogen abstraction was further investigated in a combined X-ray and neutron diffraction study on the amino acids L-serine and L-alanine, which were used as model compounds for proteins, and the nucleoside deoxythymidine (thymidine) as a model for DNA. A damage mechanism for L-serine could be found. It involves the abstraction of two hydrogen atoms, one from the hydroxyl group and one from the adjacent methylene group. Such a hydrogen abstraction results in the formation of a carbonyl group. X-ray diffraction measurements on cyano- and methylcobalamin as well as on three metal amino acid complexes, containing nickel(II) and copper(II), respectively, were conducted to investigate the contribution of X-ray induced photoreduction to global radiation damage. Results from these measurements combined with the results from L-serine, L-alanine and thymidine allowed

  16. Evidence of Chlorobenzene Natural Attenuation in Contaminated Sediments Using Compound Specific Isotope Analysis and High Resolution Pore Water Sampling

    Science.gov (United States)

    Passeport, E.; Landis, R.; Lacrampe Couloume, G.; Lutz, E. J.; Mack, E. E.; West, K.; Sherwood Lollar, B.

    2014-12-01

    Contaminated sediments can represent a significant risk for ecosystems and hinder drinking water production if contaminants discharge to surface and ground water. Understanding of contaminant fate and the potential for natural attenuation can help protect aquatic resources. In this study, the fate of chlorobenzene (MCB) and benzene was investigated in a contaminated canal sediment field site located in New Jersey, USA. Compound Specific Isotope Analysis (CSIA) was applied to sediment pore water samples collected with a peeper at high spatial resolution (3 cm) across the sediment - surface water interface (SWI). Samples were collected at three locations in canal sediments, all of which exhibited reducing redox conditions. The largest concentrations were observed in the bottommost portions of the sediment profile, with concentrations ranging from 300 to 2000 µg/L for MCB, and 16 to 180 µg/L for benzene. Conversely, concentrations were below detection limit in the surface water and in the top 6 cm of the sediment. In the zones of highest MCB concentrations, the δ13C values were -26.4 (location C) and -21.9 ‰ (location F), and became progressively more enriched in 13C while concentrations decreased, reaching -23.9 (at 12 cm below the SWI, location C) and -18.4 ‰ (at 16.5 cm below SWI, location F). Benzene was only detected in the bottom 6 cm of the sediment profiles. Benzene δ13C values were -27 (bottommost, i.e., 24 cm deep) to -29.7 ‰ (18 cm deep), in location C. Such significant isotopic enrichments in 13C (2.5 to 3.5 ‰) correlated with MCB and benzene concentration decrease are suggestive of in situ biodegradation. In addition, benzene δ13C values were systematically more depleted in 13C than MCB, suggesting that benzene found in these zones was likely produced from MCB via reductive dechlorination. This study combined for the first time CSIA with high spatial sampling resolution in surface water sediments. This setup enabled not only detection of

  17. The effect of warming and enhanced ultraviolet radiation on gender-specific emissions of volatile organic compounds from European aspen

    Energy Technology Data Exchange (ETDEWEB)

    Maja, Mengistu M., E-mail: mengistu.maja@uef.fi [University of Eastern Finland, Department of Environmental Science, P.O.Box 1627, 70211 Kuopio (Finland); Kasurinen, Anne; Holopainen, Toini [University of Eastern Finland, Department of Environmental Science, P.O.Box 1627, 70211 Kuopio (Finland); Julkunen-Tiitto, Riitta [University of Eastern Finland, Department of Biology, P.O. Box 111, 80101 Joensuu (Finland); Holopainen, Jarmo K. [University of Eastern Finland, Department of Environmental Science, P.O.Box 1627, 70211 Kuopio (Finland)

    2016-03-15

    Different environmental stress factors often occur together but their combined effects on plant secondary metabolism are seldom considered. We studied the effect of enhanced ultraviolet (UV-B) (31% increase) radiation and temperature (ambient + 2 °C) singly and in combination on gender-specific emissions of volatile organic compounds (VOCs) from 2-year-old clones of European aspen (Populus tremula L.). Plants grew in 36 experimental plots (6 replicates for Control, UV-A, UV-B, T, UV-A + T and UV-B + T treatments), in an experimental field. VOCs emitted from shoots were sampled from two (1 male and 1 female) randomly selected saplings (total of 72 saplings), per plot on two sampling occasions (June and July) in 2014. There was a significant UV-B × temperature interaction effect on emission rates of different VOCs. Isoprene emission rate was increased due to warming, but warming also modified VOC responses to both UV-A and UV-B radiation. Thus, UV-A increased isoprene emissions without warming, whereas UV-B increased emissions only in combination with warming. Warming-modified UV-A and UV-B responses were also seen in monoterpenes (MTs), sesquiterpenes (SQTs) and green leaf volatiles (GLVs). MTs showed also a UV × gender interaction effect as females had higher emission rates under UV-A and UV-B than males. UV × gender and T × gender interactions caused significant differences in VOC blend as there was more variation (more GLVs and trans-β-caryophyllene) in VOCs from female saplings compared to male saplings. VOCs from the rhizosphere were also collected from each plot in two exposure seasons, but no significant treatment effects were observed. Our results suggest that simultaneous warming and elevated-UV-radiation increase the emission of VOCs from aspen. Thus the contribution of combined environmental factors on VOC emissions may have a greater impact to the photochemical reactions in the atmosphere compared to the impact of individual factors acting alone

  18. Rapid Diagnosis of Arbovirus and Arenavirus Infections by Immunofluorescence.

    Science.gov (United States)

    1984-12-31

    1.4% Table 9 SU,"A. ’ OF EOL-A ANTIBODY SURVEY IN SERA TAKEN DURING ETHIOPIAN EPIDEMIC A. NON-EPIDEMIC AREA TESIED 227 (AWASCH, ASSAB, BLUE NILE, SIDAMO...population had group B antibody, 13% group A, 26% specific for YF, 7% specific for Zika , and 11% of the population were non-diagnostic. Similar results

  19. New indirect immunofluorescence assay as a confirmatory test for ...

    African Journals Online (AJOL)

    The IFA technique used acetone-fixed HIV-1 infected MOLT-4 cells in one spot on a Teflon coated slide and uninfected MOLT-4 cells alone in a second spot to asses non-specific fluorescence. Western blot was performed according to the instructions of the manufacturer. Results: The sensitivity and specificity of IFA based ...

  20. Development of an Analytic Method for Sulfur Compounds in Aged Garlic Extract with the Use of a Postcolumn High Performance Liquid Chromatography Method with Sulfur-Specific Detection.

    Science.gov (United States)

    Matsutomo, Toshiaki; Kodera, Yukihiro

    2016-02-01

    Garlic and its processed preparations contain numerous sulfur compounds that are difficult to analyze in a single run using HPLC. The aim of this study was to develop a rapid and convenient sulfur-specific HPLC method to analyze sulfur compounds in aged garlic extract (AGE). We modified a conventional postcolumn HPLC method by employing a hexaiodoplatinate reagent. Identification and structural analysis of sulfur compounds were conducted by LC-mass spectrometry (LC-MS) and nuclear magnetic resonance. The production mechanisms of cis-S-1-propenylcysteine (cis-S1PC) and S-allylmercaptocysteine (SAMC) were examined by model reactions. Our method has the following advantages: less interference from nonsulfur compounds, high sensitivity, good correlation coefficients (r > 0.98), and high resolution that can separate >20 sulfur compounds, including several isomers, in garlic preparations in a single run. This method was adapted for LC-MS analysis. We identified cis-S1PC and γ-glutamyl-S-allyl-mercaptocysteine in AGE. The results of model reactions suggest that cis-S1PC is produced from trans-S1PC through an isomerization reaction and that SAMC is produced by a reaction involving S-allylcysteine/S1PC and diallyldisulfide during the aging period. We developed a rapid postcolumn HPLC method for both qualitative and quantitative analyses of sulfur compounds, and this method helped elucidate a potential mechanism of cis-S1PC and SAMC action in AGE. © 2016 American Society for Nutrition.

  1. Apportioning sources of organic matter in streambed sediments: An integrated molecular and compound-specific stable isotope approach

    Energy Technology Data Exchange (ETDEWEB)

    Cooper, Richard J., E-mail: Richard.J.Cooper@uea.ac.uk [School of Environmental Sciences, University of East Anglia, Norwich Research Park, Norwich NR4 7TJ (United Kingdom); Pedentchouk, Nikolai; Hiscock, Kevin M.; Disdle, Paul [School of Environmental Sciences, University of East Anglia, Norwich Research Park, Norwich NR4 7TJ (United Kingdom); Krueger, Tobias [IRI THESys, Humboldt University, 10099 Berlin (Germany); Rawlins, Barry G. [British Geological Survey, Keyworth, Nottingham NG12 5GG (United Kingdom)

    2015-07-01

    We present a novel application for quantitatively apportioning sources of organic matter in streambed sediments via a coupled molecular and compound-specific isotope analysis (CSIA) of long-chain leaf wax n-alkane biomarkers using a Bayesian mixing model. Leaf wax extracts of 13 plant species were collected from across two environments (aquatic and terrestrial) and four plant functional types (trees, herbaceous perennials, and C{sub 3} and C{sub 4} graminoids) from the agricultural River Wensum catchment, UK. Seven isotopic (δ{sup 13}C{sub 27}, δ{sup 13}C{sub 29}, δ{sup 13}C{sub 31}, δ{sup 13}C{sub 27–31}, δ{sup 2}H{sub 27}, δ{sup 2}H{sub 29}, and δ{sup 2}H{sub 27–29}) and two n-alkane ratio (average chain length (ACL), carbon preference index (CPI)) fingerprints were derived, which successfully differentiated 93% of individual plant specimens by plant functional type. The δ{sup 2}H values were the strongest discriminators of plants originating from different functional groups, with trees (δ{sup 2}H{sub 27–29} = − 208‰ to − 164‰) and C{sub 3} graminoids (δ{sup 2}H{sub 27–29} = − 259‰ to − 221‰) providing the largest contrasts. The δ{sup 13}C values provided strong discrimination between C{sub 3} (δ{sup 13}C{sub 27–31} = − 37.5‰ to − 33.8‰) and C{sub 4} (δ{sup 13}C{sub 27–31} = − 23.5‰ to − 23.1‰) plants, but neither δ{sup 13}C nor δ{sup 2}H values could uniquely differentiate aquatic and terrestrial species, emphasizing a stronger plant physiological/biochemical rather than environmental control over isotopic differences. ACL and CPI complemented isotopic discrimination, with significantly longer chain lengths recorded for trees and terrestrial plants compared with herbaceous perennials and aquatic species, respectively. Application of a comprehensive Bayesian mixing model for 18 streambed sediments collected between September 2013 and March 2014 revealed considerable temporal variability in the

  2. Linking flow cytometric cell sorting and compound-specific 13C-analysis to determine population-specific isotopic signatures and growth rates in cyanobacteria-dominated lake plankton

    NARCIS (Netherlands)

    Pel, R.; Floris, V.; Gons, H.J.; Hoogveld, H.L.

    2004-01-01

    A novel methodology was applied to determine the 13C signatures of natural cyanobacterial and algal populations by combined compound-specific isotope ratio mass spectrometry and pyrolytic methylation-gas chromatography (Py-GC-IRMS) of the fatty acids released from phytoplankton fractions collected

  3. Reduction of quinones and nitroaromatic compounds by Escherichia coli nitroreductase A (NfsA): Characterization of kinetics and substrate specificity.

    Science.gov (United States)

    Valiauga, Benjaminas; Williams, Elsie M; Ackerley, David F; Čėnas, Narimantas

    2017-01-15

    NfsA, a major FMN-associated nitroreductase of E. coli, reduces nitroaromatic compounds via consecutive two-electron transfers. NfsA has potential applications in the biodegradation of nitroaromatic environment pollutants, e.g. explosives, and is also of interest for the anticancer strategy gene-directed enzyme prodrug therapy. However, the catalytic mechanism of NfsA is poorly characterized. Here we examined the NADPH-dependent reduction of quinones (n = 16) and nitroaromatic compounds (n = 12) by NfsA. We confirmed a general "ping-pong" reaction scheme, and preliminary rapid reaction studies of the enzyme reduction by NADPH showed that this step is much faster than the steady-state turnover number, i.e., the enzyme turnover is limited by the oxidative half-reaction. The reactivity of nitroaromatic compounds (log kcat/Km) followed a linear dependence on their single-electron reduction potential (E17), indicating a limited role for compound structure or active site flexibility in their reactivity. The reactivity of quinones was lower than that of nitroaromatics having similar E17 values, except for the significantly enhanced reactivity of 2-OH-1,4-naphthoquinones, consistent with observations previously made for the group B nitroreductase of Enterobacter cloacae. We present evidence that the reduction of quinones by NfsA is most consistent with a single-step (H-) hydride transfer mechanism. Copyright © 2016 Elsevier Inc. All rights reserved.

  4. A touchdown nucleic acid amplification protocol as an alternative to culture backup for immunofluorescence in the routine diagnosis of acute viral respiratory tract infections

    Directory of Open Access Journals (Sweden)

    Feeney Susan A

    2004-10-01

    Full Text Available Abstract Background Immunofluorescence and virus culture are the main methods used to diagnose acute respiratory virus infections. Diagnosing these infections using nucleic acid amplification presents technical challenges, one of which is facilitating the different optimal annealing temperatures needed for each virus. To overcome this problem we developed a diagnostic molecular strip which combined a generic nested touchdown protocol with in-house primer master-mixes that could recognise 12 common respiratory viruses. Results Over an 18 month period a total of 222 specimens were tested by both immunofluorescence and the molecular strip. The specimens came from 103 males (median age 3.5 y, 80 females (median age 9 y and 5 quality assurance scheme specimens. Viruses were recovered from a number of specimen types including broncho-alveolar lavage, nasopharyngeal secretions, sputa, post-mortem lung tissue and combined throat and nasal swabs. Viral detection by IF was poor in sputa and respiratory swabs. A total of 99 viruses were detected in the study from 79 patients and 4 quality control specimens: 31 by immunofluorescence and 99 using the molecular strip. The strip consistently out-performed immunofluorescence with no loss of diagnostic specificity. Conclusions The touchdown protocol with pre-dispensed primer master-mixes was suitable for replacing virus culture for the diagnosis of respiratory viruses which were negative by immunofluorescence. Results by immunofluorescence were available after an average of 4–12 hours while molecular strip results were available within 24 hours, considerably faster than viral culture. The combined strip and touchdown protocol proved to be a convenient and reliable method of testing for multiple viruses in a routine setting.

  5. 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...... and 34 induced sputum specimens were examined. All specimens were stained by five techniques: immunofluorescence using a combination of three monoclonal antibodies (from the National Institutes of Health, USA), immunofluorescence using a single monoclonal antibody (from Dakopatts), Giemsa, methenamine...... 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...

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

    Digital Repository Service at National Institute of Oceanography (India)

    Raghukumar, S.

    An immunofluorescence technique was employed to detect the thraustochytrid protist Ulkenia visurgensis (Ulken) Gaertner, originally isolated from an algal bed, in natural samples in Goa, India Collections were made monthly from September 1985...

  7. 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.

  8. Variations of attractors and wavelet spectra of the immunofluorescence distributions for women in the pregnant period

    Science.gov (United States)

    Galich, Nikolay E.

    2008-07-01

    Communication contains the description of the immunology data treatment. New nonlinear methods of immunofluorescence statistical analysis of peripheral blood neutrophils have been developed. We used technology of respiratory burst reaction of DNA fluorescence in the neutrophils cells nuclei due to oxidative activity. The histograms of photon count statistics the radiant neutrophils populations' in flow cytometry experiments are considered. Distributions of the fluorescence flashes frequency as functions of the fluorescence intensity are analyzed. Statistic peculiarities of histograms set for women in the pregnant period allow dividing all histograms on the three classes. The classification is based on three different types of smoothing and long-range scale averaged immunofluorescence distributions, their bifurcation and wavelet spectra. Heterogeneity peculiarities of long-range scale immunofluorescence distributions and peculiarities of wavelet spectra allow dividing all histograms on three groups. First histograms group belongs to healthy donors. Two other groups belong to donors with autoimmune and inflammatory diseases. Some of the illnesses are not diagnosed by standards biochemical methods. Medical standards and statistical data of the immunofluorescence histograms for identifications of health and illnesses are interconnected. Peculiarities of immunofluorescence for women in pregnant period are classified. Health or illness criteria are connected with statistics features of immunofluorescence histograms. Neutrophils populations' fluorescence presents the sensitive clear indicator of health status.

  9. Compound-Specific δ¹⁵N and δ¹³C Analyses of Amino Acids for Potential Discrimination between Organically and Conventionally Grown Wheat.

    Science.gov (United States)

    Paolini, Mauro; Ziller, Luca; Laursen, Kristian Holst; Husted, Søren; Camin, Federica

    2015-07-01

    We present a study deploying compound-specific nitrogen and carbon isotope analysis of amino acids to discriminate between organically and conventionally grown plants. We focused on grain samples of common wheat and durum wheat grown using synthetic nitrogen fertilizers, animal manures, or green manures from nitrogen-fixing legumes. The measurement of amino acid δ(15)N and δ(13)C values, after protein hydrolysis and derivatization, was carried out using gas chromatography-combustion-isotope ratio mass spectrometry (GC-C-IRMS). Our results demonstrated that δ(13)C of glutamic acid and glutamine in particular, but also the combination of δ(15)N and δ(13)C of 10 amino acids, can improve the discrimination between conventional and organic wheat compared to stable isotope bulk tissue analysis. We concluded that compound-specific stable isotope analysis of amino acids represents a novel analytical tool with the potential to support and improve the certification and control procedures in the organic sector.

  10. Carbon sources in suspended particles and surface sediments from the Beaufort Sea revealed by molecular lipid biomarkers and compound-specific isotope analysis

    OpenAIRE

    I. Tolosa; S. Fiorini; B. Gasser; J. Martín; J. C. Miquel

    2013-01-01

    Molecular lipid biomarkers (hydrocarbons, alcohols, sterols and fatty acids) and compound-specific isotope analysis of suspended particulate organic matter (SPM) and surface sediments of the Mackenzie Shelf and slope (southeast Beaufort Sea, Arctic Ocean) were studied in summer 2009. The concentrations of the molecular lipid markers, characteristic of known organic matter sources, were grouped and used as proxies to evaluate the relative importance of fresh algal, detrital a...

  11. Comparison of indirect immunofluorescence with agar gel immunodiffusion for the diagnosis of bluetongue virus infection.

    Science.gov (United States)

    Arita, G M; Gatti, M S; Germano, P M; Pestana-de-Castro, A F

    1992-01-01

    1. Sera from 190 cows and from 72 sheep were examined to compare the results obtained with the agar gel immunodiffusion (AGID) and indirect immunofluorescence (IIF) tests for the diagnosis of bluetongue (BT) disease. 2. In the AGID test, 96 of 190 (50.5%) cattle serum samples and 38 of 72 (52.7%) sheep serum samples were positive, for a total of 134 out of 262 (51.1%) sera. In the IIF test, 98 of 190 (51.6%) cattle serum samples and 39 of 72 (54.2%) sheep serum samples were positive, for a total of 137 out of 262 (52.3%) sera. 3. The fluorescence of the IIF test presented a granular cytoplasmic aspect, which in some cells was observed only on the cell membranes. 4. Statistical analysis of the data showed close agreement between the two techniques, regardless of the kind of sera examined. The IIF test showed high sensitivity (93.8% and 92.1%), specificity (91.4% and 88.2%) and positive (91.8% and 89.7%) and negative (93.48% and 90.9%) predictive values for cattle serum and sheep serum, respectively. 5. The results obtained with IIF were comparable to those obtained with the AGID test, indicating that both techniques can be used routinely in epidemiologic studies of BT. However, the IIF offers the additional advantages that it can be used for antibody quantification and for the detection of viral antigens in BT-infected cell lines.

  12. Immunofluorescent localization of Tamm-Horsfall mucoprotein in human kidney

    Science.gov (United States)

    McKenzie, J. K.; McQueen, E. G.

    1969-01-01

    A fluorescein-labelled antiserum to human Tamm-Horsfall mucoprotein applied to frozen human kidney sections gave strong specific labelling, mainly of cells of tubules in the outer medulla. By comparison with adjacent serial sections stained for alkaline phosphatase and succinic dehydrogenase, it is suggested that material reacting immunologically as Tamm-Horsfall muco-protein is found particularly in the cells of the ascending limb of the loop of Henle and the macula densa segment of the distal tubule. Images PMID:4891482

  13. Melasma: a clinical, light microscopic, ultrastructural, and immunofluorescence study.

    Science.gov (United States)

    Sanchez, N P; Pathak, M A; Sato, S; Fitzpatrick, T B; Sanchez, J L; Mihm, M C

    1981-06-01

    Melasma is an acquired brown hypermelanosis of the face. Although it is thought that melasma is associated with multiple etiologic factors (pregnancy, gastric, racial, and endocrine), one of the primary causes of its exacerbation appears to be exposure to sunlight. Three patterns of melasma are recognized clinically: (1) a centrofacial pattern, (2) a malar pattern, and (3) a mandibular pattern. Examination of patients with Wood's light (320--400 nm) is useful in classifying the specific type of melasma in correlation with the localization of pigment granules (melanosomes) in the epidermis and dermis. Four types of melasma are described on the basis of Wood's light examination: (1) an epidermal type, (2) a dermal type, (3) a mixed type, and (4) a fourth type, described in patients of dark complexion, in which the lesions, for lack of contrast, are not discernible on Wood's light examination, perhaps due to the increased number of melanosomes in the normal skin of black individuals. Light, histochemical, and electron microscopic studies revealed an increase in number and activity of type-specific melanocytes which appeared to be engaged in increased formation, melanization, and transfer of pigment granules (melanosomes) to the epidermis as well as to the dermis. The melanocyte seems to undergo a functional alteration brought about by a combination of multiple factors, including persistent sun exposure, hormonal factors, and genetic predisposition.

  14. 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.

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

    Science.gov (United States)

    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.

  16. 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...... but only 71% sensitivity. The Giardia ELISA correlated well with IF (sensitivity 100%, specificity 96%) and was capable of detecting animal specific Giardia duodenalis genotypes. Visual interpretation appeared appropriate for assessment of ELISA results. The proportion of positive samples and possible...... 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...

  17. Titanium dioxide as chemo-affinity chromatographic sorbent of biomolecular compounds - Applications in acidic modification-specific proteomics

    DEFF Research Database (Denmark)

    Engholm-Keller, Kasper; Larsen, Martin R

    2011-01-01

    Characterization of the biomolecules involved in molecular processes occurring in biological systems such as the human cell remains central to biology, biotechnology, and medicine. One of the preferred methods of selectively purifying specific classes of biomolecules from complex biological matri...

  18. Organ-specific distribution of phenolic compounds in bilberry (Vaccinium myrtillus) and 'northblue' blueberry (Vaccinium corymbosum x V. angustifolium).

    Science.gov (United States)

    Riihinen, Kaisu; Jaakola, Laura; Kärenlampi, Sirpa; Hohtola, Anja

    2008-09-01

    Blueberries and bilberries are recognized as some of the best sources of flavonoids, especially anthocyanins. The contents of flavonoids (anthocyanins, proanthocyanidins, flavonols) and hydroxycinnamic acids in the flower, fruit skin and pulp, leaf and rhizome of bilberry and the blueberry cultivar 'Northblue' were analyzed using high-performance liquid chromatography combined with diode-array detection. The most striking difference in the fruits was the predominance of hydroxycinnamic acids in blueberry, whereas in bilberry the anthocyanin content was much higher, particularly in the pulp. Differences in flavonoid contents of fruits were already apparent at the flower stage. Bilberry and blueberry leaves both contained high amounts of proanthocyanidins, flavonols and hydroxycinnamic acids. Blueberry rhizomes accumulated high amounts of hydroxycinnamic acids. All plant parts of bilberry and blueberry are potential sources of phenolic compounds for use either as dietary botanicals or by the pharmaceutical industry. Copyright © 2008 Elsevier Ltd. All rights reserved.

  19. Bulk and Compound Specific Isotope Analysis Provide Insights into Habitat Use and Diet Variability Among Female Sperm Whales (Physeter macrocephalus) from the Eastern Tropical Pacific

    Science.gov (United States)

    Zupcic-Moore, J. R.; Ruiz-Cooley, R. I.; McCarthy, M. D.; Koch, P. L.

    2016-02-01

    Sperm whales (Physeter macrocephalus) have a strong social structure and females form sympatric clans that span thousands of kilometers and encompass thousands of whales. We investigated variability in foraging habitat and diet through the life history of ten female sperm whales from the Eastern Tropical Pacific (ETP), using coupled bulk and compound specific amino acid stable isotope measurements made on annual tooth growth layers. We identified three broad temporal patterns in annual records of bulk nitrogen (δ15N) and carbon (δ13C) isotope values in sub-groups of individuals: 1) constant, 2) increasing, and 3) decreasing isotopic values between ca.1930 and 1960. These different isotopic patterns suggest that whales sampled may belong to different clans. In addition, average bulk δ15N and δ13C values differed among individuals and appear to be group-specific. This suggests that whales from different sub-groups foraged in different habitats and/or had differing diets. To differentiate between these interpretations, we conducted compound specific amino acid isotope analysis on dentin samples from six whales. Amino acid proxies for baseline δ15N values indicate that bulk variability among whales is due to δ15N differences at the base of the food web, indicating different feeding areas. The δ13C values of essential amino acids, which are positively correlated to bulk δ13C values, support this conclusion. Together, the compound specific and bulk annual records suggest that whales with higher isotopic values likely foraged in regions closer to the coast throughout their lives, while whales with lower isotopic values likely foraged further offshore. Overall, our data suggest clear spatial segregation of clans within a similar ocean region. Since molecular analysis cannot yet distinguish genetic variability among whale clans, we suggest that stable isotope analysis can help to identify clans and their foraging ecology, based on habitat use.

  20. Analytical model for site-specific isotope fractionation in 13C during sorption: determination by isotopic 13C NMR spectrometry with vanillin as model compound.

    Science.gov (United States)

    Höhener, Patrick; Silvestre, Virginie; Lefrançois, Anaïs; Loquet, Denis; Botosoa, Eliot P; Robins, Richard J; Remaud, Gérald S

    2012-04-01

    The aim of this study was to conceive a reactive transport model capable of providing quantitative site-specific enrichment factors for fractionation in (13)C isotopic content during sorption. As test compound the model treats vanillin, for which the (13)C isotopic content at natural abundance at each of the 8 carbon positions can be measured by quantitative (13)C nuclear magnetic resonance spectrometry. This technique determines the isotope ratios with a resolution better than ±1‰ (0.1%) at each carbon position. Site-specific isotope fractionations were recorded in chromatography column experiments with silica RP-18 as stationary phase. The one dimensional reactive transport model accounted for the sorption/desorption behavior of 8 individual (13)C-isotopomers and one (12)C-isotopomer of vanillin and reproduced satisfactorily the bulk (average over the whole compound) fractionation observed during elution. After model calibration, the enrichment factors were fitted for each carbon site where a significant fractionation was recorded. To show the interest of such a transport model for environmental studies, the model, extended to three dimensions, was exploited to simulate reactive transport in an aquifer. These results show that significant (13)C isotope fractionation is expected for 4 out of 8 (13)C-isotopomers in vanillin, and illustrate that bulk isotope ratios measured by conventional compound specific isotope analysis and mass spectrometry would hardly document significant isotope fractionations in vanillin. It is concluded that modeling of site-specific isotope ratios in molecules is a priori feasible and may help to quantify unknown processes in the environment. Copyright © 2011 Elsevier Ltd. All rights reserved.

  1. Compound specific stable isotopes as probes for distinguishing the sources of biomolecules in terrestrial and extraterrestrial materials

    Science.gov (United States)

    Engel, M. H.; Macko, S. A.

    2003-04-01

    Life on Earth consists of orderly arrangements of several key types of organic compounds (amino acids, sugars, fatty acids, nucleic bases) that are the building blocks of proteins, carbohydrates, lipids and nucleotides. Subsequent to death, macromolecules are commonly broken down to their molecular constituents or other similar scale components. Thus, in ancient terrestrial and extraterrestrial materials, it is far more likely to expect the presence of simple compounds such as amino acids rather than the proteins from which they were possibly derived. Given that amino acids, for example, are common components of all extinct and extant organisms, the challenge has been to develop methods for distinguishing their sources. Stable isotopes are powerful probes for determining the origins of organic matter. Amino acid constituents of all organisms on Earth exhibit characteristic stable isotope compositions owing to fractionations associated with their biosynthesis. These fractionations are distinct from those observed for amino acids formed by abiotic processes. Thus it should be possible to use isotopes as probes for determining whether amino acids in ancient rocks on Earth are biotic or abiotic, based on their relative isotopic compositions. Also, owing to differences in the isotope compositions of precursors, amino acids in extraterrestrial materials such as carbonaceous meteorites are moderately to substantially enriched in the heavy isotopes of C, N and H relative to terrestrial amino acids. Assuming that the isotope compositions of the gaseous components of, for example, the Martian atmosphere were distinct from Earth at such time when organic molecules may have formed, it should be possible to distinguish these components from terrestrial contaminants by determining their isotope compositions and/or those of their respective enantiomers. Also, if life as we know it existed on another planet such as Mars, fractionations characteristic of biosynthesis should be

  2. Probing the specificity of polyurethane foam as a 'solid-phase extractant': Extractability-governing molecular attributes of lipophilic phenolic compounds.

    Science.gov (United States)

    Han, Jie; Cao, Zhi; Qiu, Wei; Gao, Wei; Hu, Jiangyong; Xing, Baoshan

    2017-09-01

    The long-proposed use of open-cell polyurethane foam (PUF) as a convenient 'solid-phase extractant' for aqueous organic compounds has been hindered by a critical lack of understanding on the underlying specificity of its extraction mechanism. In this work, we tasked ourselves to understand the hierarchy of molecular structure, properties, and partitioning characteristics of compounds in PUF and aqueous phase by targeting lipophilic phenolic compounds (LPCs) as a group of primary targets for PUF extraction. Using six structurally related bisphenol analogs as comparative probes, we identified molecular lipophilicity and H-bond acidity as key molecular attributes that governed their extractability by PUF. Molecular modeling study on H-bonding interactions between PUF surrogates, bisphenols, and water molecules elucidated the governing effect of H-bond acidity in the binding affinity of guest molecules onto PUF lone-pair donors. A holistic view must be adopted when assessing the extractability of LPCs with reactive lone-pair donors e.g. bisphenol S which forms multidentate H-bond adducts with water molecules. We validated our theory on two model groups of monofunctional LPCs, alkylphenols and chlorophenols, with the observation that the presence of a second proton-donating moiety dramatically enhanced the extractability of bisphenol molecules. The specificity of PUF rendered it selective towards compounds with correlating molecular attributes against other structural analogs and co-existing matrix organics. For LPCs, the PUF macromolecular structure can be conceptualized as a flexible 'molecular zipper network' that is most affinitive towards nonionic, permeable, and lipophilic guest molecules with multiple reactive proton donors. Copyright © 2017 Elsevier B.V. All rights reserved.

  3. Temporal Variability of Source-Specific Solvent-Extractable Organic Compounds in Coastal Aerosols over Xiamen, China

    Directory of Open Access Journals (Sweden)

    Shuqin Tao

    2017-02-01

    Full Text Available This study describes an analysis of ambient aerosols in a southeastern coastal city of China (Xiamen in order to assess the temporal variability in the concentrations and sources of organic aerosols (OA. Molecular-level measurements based on a series of solvent extractable lipid compounds reveal inherent heterogeneity in OA, in which the concentration and relative contribution of at least three distinct components (terrestrial plant wax derived, marine/microbial and fossil fuel derived organic matter (OM exhibited distinct and systematic temporal variability. Plant wax lipids and associated terrestrial OM are influenced by seasonal variability in plant growth; marine/microbial lipids and associated marine OM are modulated by sea spill and temperature change, whereas fossil fuel derived OM reflects the anthropogenic utilization of fossil fuels originated from petroleum-derived sources and its temporal variation is strongly controlled by meteorological conditions (e.g., the thermal inversion layer, which is analogous to other air organic pollutions. A comparative study among different coastal cities was applied to estimate the supply of different sources of OM to ambient aerosols in different regions, where it was found that biogenic OM in aerosols over Xiamen was much lower than that of other cities; however, petroleum-derived OM exhibited a high level of contribution with a higher concentration of unresolved complex matters (UCM and higher a ratio between UCM and resolved alkanes (UCM/R.

  4. Specific phenolic compounds and sensory properties of a new dealcoholized red wine with pomegranate (Punica granatum L.) extract.

    Science.gov (United States)

    Tárrega, Maria Amparo; Varela, Paula; Fromentin, Emilie; Feuillère, Nicolas; Issaly, Nicolas; Roller, Marc; Sanz-Buenhombre, Marisa; Villanueva, Sonia; Moro, Carlos; Guadarrama, Alberto; Fiszman, Susana

    2014-09-01

    The pomegranate (Punica granatum L.) fruit has a long history of human consumption and possesses notable antioxidant and cardiovascular properties. This work evaluated the feasibility to provide a new functional beverage based on a dealcoholized red wine matrix supplemented by a pomegranate extract. The potential bioactive compounds in the pomegranate extract, punicalagin A and B and ellagic acid, were analyzed during the downstream process in order to evaluate the functional dose in the final beverage. The addition of pomegranate extract to the dealcoholized red wine resulted in a product with more intense yeast odor, acidity, yeast flavor, and astringency and with a less intense berry flavor. Consumer acceptance of the product was also investigated and the results revealed the existence of a niche of consumers willing to consume dealcoholized wine enriched with pomegranate extract. After tasting, 50% and 40% of those consumers initially interested by this product concept declared to be interested to purchase the control sample and the functional beverage, respectively. The daily consumption of two servings of 250 mL of this new pomegranate-enriched dealcoholized wine provides 82 mg of total ellagitannins, corresponding to the sum of punicalagin A and B and ellagic acid. © The Author(s) 2013 Reprints and permissions: sagepub.co.uk/journalsPermissions.nav.

  5. Production of anti-Cryptosporidium polyclonal antibodies and standardization of direct immunofluorescence for detecting oocysts in water

    Directory of Open Access Journals (Sweden)

    Silvia Cristina Osaki

    2011-10-01

    Full Text Available INTRODUCTION: The production of anti-Cryptosporidium polyclonal antibodies and its use in direct immunofluorescence assays to determine the presence of Cryptosporidium in water are described in the present work. METHODS: Two rabbits were immunized with soluble and particulate antigens from purified Cryptosporidium oocysts. The sera produced were prepared for immunoglobulin G extraction, which were then purified and conjugated with fluorescein isothiocyanate (FITC. Slides containing known amounts of oocysts were prepared to determine the sensitivity of the technique. To test the specificity, slides containing Giardia duodenalis cysts were prepared. RESULTS: The conjugate was successfully used in water samples experimentally contaminated with Cryptosporidium oocysts, and it was possible to detect up to five oocysts/spot, corresponding to contamination of 250 oocysts/mL. CONCLUSIONS: The three immunizations performed in the rabbits were enough to produce antibodies against Cryptosporidium, the standard direct immunofluorescence assay permitted the detection of five oocysts in 20% of the samples, and no cross-reaction with Giardia duodenalis cysts occurred.

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

    Science.gov (United States)

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

    1997-02-01

    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 the urodele Triturus carnifex. After biocytin insertions in the pars intermedia, neurons became retrogradely labelled in the suprachiasmatic hypothalamus and the locus coeruleus of the brainstem in both species. Some scattered biocytin-labelled neurons were observed in the preoptic area. Moreover, working on the same sections, immunofluorescence revealed a number of codistributions and, in some cases, colocalization in the same neurons of biocytin labellings and immunopositivity for (1) tyrosine hydroxylase in the suprachiasmatic hypothalamus and the locus coeruleus of Rana and Triturus, (2) gamma-aminobutyric acid in the suprachiasmatic hypothalamus of Rana and Triturus and (3) neuropeptide Y in the suprachiasmatic hypothalamus of Rana. The specificity of such colocalizations was fully confirmed using dual-channel confocal laser scanning microscopy analysis.

  7. 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...

  8. Effectiveness of PCR and Immunofluorescence Techniques for Detecting Human Cytomegalovirus in Blood and Bronchoalveolar Lavage Fluid.

    Science.gov (United States)

    Roży, A; Duk, K; Szumna, B; Skrońska, P; Gawryluk, D; Chorostowska-Wynimko, J

    Current diagnostic methods allow a rapid and reliable detection of active human cytomegalovirus (hCMV) infection by identifying the presence of pp65 CMV antigen or CMV DNA in peripheral blood and affected organs. The goal of this study was to evaluate the effectiveness of CMV detection in blood and organ-specific biological material, such as bronchoalveolar lavage fluid (BALF), by comparing two standard diagnostic methods, immunofluorescence (IF) and the real-time polymerase chain reaction (PCR). We evaluated 25 patients with concomitant respiratory disease who were referred to our hospital for diagnosis due to suspected acute CMV infection. The presence of hCMV was concomitantly evaluated by IF and PCR in 16 peripheral blood samples. In two patients, we observed positive results for both IF and PCR, and in two other patients the results were discordant. Of 11 patients, CMV DNA was detected in six BALF samples, and in one blood plasma sample. Real-time PCR detected CMV DNA in 54.6 % of BALF samples and 12.0 % of blood samples, while indirect IF testing confirmed antigenemia in 12.5 % of blood samples. The results from our study suggest that the IF method is as effective as PCR for detecting an ongoing CMV infection in blood samples. However, real-time PCR was much more effective at detecting CMV DNA in BALF compared to blood samples. Our results suggest that the biological material being tested during CMV diagnosis should be derived directly from the virally infected organ(s).

  9. Immunofluorescence tests for HIV antibody and their value as confirmatory tests.

    Science.gov (United States)

    van der Groen, G; Vercauteren, G; Piot, P

    1987-08-01

    The enzyme-linked immunosorbent assay (ELISA) is currently being used as a sensitive screening test for HIV antibody. The immunoblot assay (IBA) and indirect immunofluorescence (IF) techniques are two recommended confirmation tests for EIA-positive sera. An indirect IF test has been developed by various laboratories using acetone fixed mixtures of uninfected and HIV-infected cells, which facilitated the reading, since nonspecific reactions were easily differentiated from specific staining. Similar results have been obtained with H9-, CEM-, and HUT78-HIV-infected and uninfected cells. Anti-nuclear antibodies and auto-antibodies resulting in false-positive EIA results, could easily be differentiated by the IF test. Aspecific fluorescence can be removed by absorption of the specimens with non-infected cells. However, IF is not suitable for the screening of large series of specimens. IF is especially well suited for quantitative analysis of serum antibody levels. Whereas serum antibody titers rise initially after infection, they decrease as AIDS develops. Heat inactivation of sera did not affect reactivity in IF, in contrast to a high rate of false-positive results obtained with heat inactivated sera in some ELISAs. A well characterized serum from an AIDS patient can be used to perform IF in order to monitor HIV infection of susceptible cells. It has been claimed that titers of neutralizing antibodies significantly correlate with the levels of IF anti-HIV antibodies. An overall correlation of 99% between IF and IBA was reported by different laboratories, when HIV ELISA-reactive European and North American sera were tested. The concordance with IBA was 97% when HIV ELISA-reactive African sera were tested.(ABSTRACT TRUNCATED AT 250 WORDS)

  10. Preparation of organic-silica hybrid boronate affinity monolithic column for the specific capture and separation of cis-diol containing compounds.

    Science.gov (United States)

    Li, Qianjin; Lü, Chenchen; Li, Hengye; Liu, Yunchun; Wang, Heye; Wang, Xin; Liu, Zhen

    2012-09-21

    A new boronate-silica hybrid monolithic column was prepared using a one-pot approach with 3-acrylamidophenylboronic acid (AAPBA) as the boronate affinity ligand. The AAPBA-silica monolith exhibited several attractive advantages. First, it is highly hydrophilic, providing excellent specificity and avoiding the presence of organic solvent in the mobile phase. Second, due to its large surface area, it exhibited a high binding capacity, 49.5 μmol/mL, the highest among the boronate affinity monolithic columns appeared in the literature. Third, the monolith can bind with cis-diol containing compounds at pH as low as 6.5, which not only avoids the use of basic pH conditions at which the silica monolith may hydrolysis but also facilitates the applications to wider sample range. Finally, the hybrid monolithic column exhibited apparent secondary separation capability, which allows for two-dimensional (2D) separation of cis-diol compounds in a single column. Due to these merits, the AAPBA-silica hybrid monolithic column can be a promising separation medium for the analysis of cis-diol containing compounds. Copyright © 2012 Elsevier B.V. All rights reserved.

  11. The Mechanism of Melanocytes-Specific Cytotoxicity Induced by Phenol Compounds Having a Prooxidant Effect, relating to the Appearance of Leukoderma

    Directory of Open Access Journals (Sweden)

    Takeshi Nagata

    2015-01-01

    Full Text Available Specific phenol compounds including rhododendrol (RD, a skin-brightening ingredient in cosmetics, are reported to induce leukoderma, inducing a social problem, and the elucidation of mechanism of leukoderma is strongly demanded. This study investigated the relationship among the cytotoxicities of six phenol compounds on B16F10 melanoma cells and HaCaT keratinocytes and generated reactive oxygen species (ROS. As a result, the cytotoxicity of RD on B16F10 cells was higher than that on HaCaT cells, and RD significantly increased intracellular ROS and hydrogen peroxide (H2O2 levels in B16F10 cells. Furthermore, although raspberry ketone (RK, RD derivative, also increased intracellular ROS in B16F10 cells, increase in ROS was suppressed by disodium dihydrogen ethylenediaminetetraacetate dehydrate (EDTA. The amounts of increased ROS with RK in HaCaT cells without melanocyte were further increased by tyrosinase. Therefore, tyrosinase, a metalloprotein having copper, was speculated to be one of causative agents allowing phenol compounds to work as a prooxidant. Hydroxyl radical was generated by adding a mixture of tyrosinase and H2O2 to RD, and the amount of the radical was further increased by UVB, indicating that RD cytotoxicity was caused by intracellularly increased ROS, which possibly related to phenol induced prooxidants.

  12. Biogenic volatile organic compound (BVOC) emissions from forested areas in Turkey: Determination of specific emission rates for thirty-one tree species

    Energy Technology Data Exchange (ETDEWEB)

    Aydin, Yagmur Meltem; Yaman, Baris; Koca, Husnu; Dasdemir, Okan; Kara, Melik; Altiok, Hasan; Dumanoglu, Yetkin; Bayram, Abdurrahman [Department of Environmental Engineering, Faculty of Engineering, Dokuz Eylul University, Tinaztepe Campus, Buca, Izmir (Turkey); Tolunay, Doganay [Department of Soil Science and Ecology, Faculty of Forestry, Istanbul University, Bahcekoy, Istanbul (Turkey); Odabasi, Mustafa [Department of Environmental Engineering, Faculty of Engineering, Dokuz Eylul University, Tinaztepe Campus, Buca, Izmir (Turkey); Elbir, Tolga, E-mail: tolga.elbir@deu.edu.tr [Department of Environmental Engineering, Faculty of Engineering, Dokuz Eylul University, Tinaztepe Campus, Buca, Izmir (Turkey)

    2014-08-15

    Normalized biogenic volatile organic compound (BVOC) emission rates for thirty one tree species that cover the 98% of national forested areas in Turkey were determined. Field samplings were performed at fourteen different forested areas in Turkey using a specific dynamic enclosure system. The selected branches of tree species were enclosed in a chamber consisted of a transparent Nalofan bag. The air-flows were sampled from both inlet and outlet of the chamber by Tenax-filled sorbent tubes during photosynthesis of trees under the presence of sunlight. Several environmental parameters (temperature, humidity, photosynthetically active radiation-PAR, and CO{sub 2}) were continuously monitored inside and outside the enclosure chamber during the samplings. Collected samples were analyzed using a gas chromatography mass spectrometry (GC/MS) system equipped with a thermal desorber (TD). Sixty five BVOCs classified in five major groups (isoprene, monoterpenes, sesquiterpenes, oxygenated sesquiterpenes, and other oxygenated compounds) were analyzed. Emission rates were determined by normalization to standard conditions (1000 μmol/m{sup 2} s PAR and 30 °C temperature for isoprene and 30 °C temperature for the remaining compounds). In agreement with the literature, isoprene was mostly emitted by broad-leaved trees while coniferous species mainly emitted monoterpenes. Several tree species such as Sweet Chestnut, Silver Lime, and European Alder had higher monoterpene emissions although they are broad-leaved species. High isoprene emissions were also observed for a few coniferous species such as Nordmann Fir and Oriental Spruce. The highest normalized total BVOC emission rate of 27.1 μg/g h was observed for Oriental Plane while South European Flowering Ash was the weakest BVOC emitter with a total normalized emission rate of 0.031 μg/g h. Monoterpene emissions of broad-leaved species mainly consisted of sabinene, limonene and trans-beta-ocimene, while alpha-pinene, beta

  13. 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.

  14. The CSSIAR v.1.00 Software: A new tool based on SIAR to assess soil redistribution using Compound Specific Stable Isotopes

    Science.gov (United States)

    Sergio, de los Santos-Villalobos; Claudio, Bravo-Linares; dos Anjos Roberto, Meigikos; Renan, Cardoso; Max, Gibbs; Andrew, Swales; Lionel, Mabit; Gerd, Dercon

    Soil erosion is one of the biggest challenges for food production around the world. Many techniques have been used to evaluate and mitigate soil degradation. Nowadays isotopic techniques are becoming a powerful tool to assess soil apportionment. One of the innovative techniques used is the Compound Specific Stable Isotopes (CSSI) analysis, which has been used to track down sediments and specify their sources by the isotopic signature of δ13 C in specific fatty acids. The application of this technique on soil apportionment has been recently developed, however there is a lack of user-friendly Software for data processing and interpretation. The aim of this article is to introduce a new open source tool for working with data sets generated by the use of the CSSI technique to assess soil apportionment, called the CSSIARv1.00 Software

  15. The CSSIAR v.1.00 Software: A new tool based on SIAR to assess soil redistribution using Compound Specific Stable Isotopes

    Directory of Open Access Journals (Sweden)

    de los Santos-Villalobos Sergio

    2017-01-01

    Full Text Available Soil erosion is one of the biggest challenges for food production around the world. Many techniques have been used to evaluate and mitigate soil degradation. Nowadays isotopic techniques are becoming a powerful tool to assess soil apportionment. One of the innovative techniques used is the Compound Specific Stable Isotopes (CSSI analysis, which has been used to track down sediments and specify their sources by the isotopic signature of δ13C in specific fatty acids. The application of this technique on soil apportionment has been recently developed, however there is a lack of user-friendly Software for data processing and interpretation. The aim of this article is to introduce a new open source tool for working with data sets generated by the use of the CSSI technique to assess soil apportionment, called the CSSIARv1.00 Software

  16. Characterization of volatile organic compounds (VOCs in Asian and north American pollution plumes during INTEX-B: identification of specific Chinese air mass tracers

    Directory of Open Access Journals (Sweden)

    A. J. Weinheimer

    2009-07-01

    Full Text Available We present results from the Intercontinental Chemical Transport Experiment – Phase B (INTEX-B aircraft mission conducted in spring 2006. By analyzing the mixing ratios of volatile organic compounds (VOCs measured during the second part of the field campaign, together with kinematic back trajectories, we were able to identify five plumes originating from China, four plumes from other Asian regions, and three plumes from the United States. To identify specific tracers for the different air masses we characterized their VOC composition and we compared their background levels with those obtained during the 2004 INTEX-A mission. The Chinese and other Asian air masses were significantly enhanced in carbonyl sulfide (OCS and methyl chloride (CH3Cl, while all CFC replacement compounds were elevated in US plumes, particularly HFC-134a.

    Although elevated mixing ratios of Halon-1211 were measured in some Chinese plume samples, several measurements at background levels were also observed. After analyzing the VOC distribution and correlations within the Chinese pollution plumes and applying principal component analysis (PCA, we suggest the use of a suite of species, rather than a single gas, as specific tracers of Chinese air masses (namely OCS, CH3Cl, 1,2-dichloroethane, ethyl chloride, and Halon-1211. In an era of constantly changing halocarbon usage patterns, this suite of gases best reflects new emission characteristics from China.

  17. The development of a highly specific radiochemical compound based on labeled 99mtc recombinant molecules for targeted imaging of cells with the overexpression of Her-2 / neu

    Directory of Open Access Journals (Sweden)

    Olga D. Bragina

    2017-01-01

    Full Text Available Currently, there is a urgent need to search for new diagnostic methods that allow us to reveal malignant tumors with the overexpression of Her-2/neu with high accuracy. In recent years radioisotope methods have been actively developing to identify specific tumor targets, with antibodies being the “targeting” module.The purpose of the study. Creation of a chemically stable radiochemical compound for the imaging of cells with the overexpression of Her-2/neu.Materials and methods. The study was conducted using two human adenocarcinoma cell lines with expression (BT-474 and without expression (MCF-7 Her-2/neu. The specificity of the binding of the test complex with Her-2/neu receptor was determined by direct radiometric and planar scintigraphy. To evaluate the differences in quantitative characteristics between the groups a non-parametric Mann – Whitney test was used.Results. The yield of the labeled complex was more than 91% and the radiochemical frequency was more than 94%. When performing a visual scintigraphic evaluation, a much higher accumulation rate of the studied radiopharmaceutical preparation (RFP was observed in the culture of cells with overexpression of the surface Her-2/neu receptor. Direct radiometric results also demonstrated a higher accumulation of RFPs in the human BT-474 mammary adenocarcinoma cell line with Her-2/neu overexpression in comparison with the control group.Conclusion. Preclinical studies demonstrated high stability of the test compound, as well as its accumulation in the group of cells with Her-2/neu overexpression

  18. Experimental and epidemiological evidence on non-organ specific cancer preventive effect of Korean ginseng and identification of active compounds

    Energy Technology Data Exchange (ETDEWEB)

    Yun, T.-K

    2003-03-01

    Panax ginseng C.A. Meyer has been the most highly recognized medicinal herb in the Orient. The prolonged administration of red ginseng extract significantly inhibits the incidence of hepatoma and also proliferation of pulmonary tumors induced by aflatoxin B{sub 1} and urethane. Statistically significant anticarcinogenic effects were in aged or heat treated extracts of ginseng and red ginseng made by steaming in a 9 weeks medium-term anticarcinogenicity test using benzo[a]pyrene. In case-control studies, odds ratios (OR) of the cancer of lip, oral cavity and pharynx, larynx, lung, esophagus, stomach, liver, pancreas, ovary, and colorectum were significantly reduced. As to the type of ginseng, the ORs for cancer were reduced in user of fresh ginseng extract intakers, white ginseng extract, white ginseng powder, and red ginseng. In a cohort study with 5 years follow-up conducted in a ginseng cultivation area, ginseng users had a decreased relative risk (RR) compared with non-users. The relative risks (RRs) of ginseng users were decreased in gastric cancer and lung cancer. These findings strongly suggest that Panax ginseng C.A. Meyer cultivated in Korea has non-organ specific cancer preventive effects against various cancers. To investigate the active components for cancer prevention, several fractions of fresh and red ginseng and four semi-synthetic ginsenoside Rh{sub 1}, Rh{sub 2}, Rg{sub 3} and Rg{sub 5}, the major saponin components in red ginseng, were prepared among the ginsenosides. By using Yun's model, Rg{sub 3} and Rg{sub 5} showed statistically significant reduction of lung tumor incidence and Rh{sub 2} had a tendency to decrease the incidence. In conclusion, these results strongly suggested that Panax ginseng C.A. Meyer cultivated in Korea is a non-organ specific cancer preventive against human cancers and also indicated that the anticarcinogenicity or human cancer preventive effect of Panax ginseng is due to ginsenoside Rg{sub 3}, Rg{sub 5} and Rh

  19. Comparison of immunofluorescence and enzyme-linked immunosorbent assays for the serology of hantavirus infections.

    NARCIS (Netherlands)

    J. Groen (Jan); G. van der Groen (Guido); G. Hoofd; A.D.M.E. Osterhaus (Albert)

    1989-01-01

    textabstractThree enzyme-linked immunosorbent assay (ELISA) systems based upon different principles were developed for the serology of Hantaan virus infections and compared with an indirect immunofluorescence assay (IFA). The indirect IFA was carried out with gamma-irradiated Hantaan virus-infected

  20. Immune diagnosis of pure ocular mucous membrane pemphigoid : indirect immunofluorescence versus immunoblot

    NARCIS (Netherlands)

    Jonkman, Marcell F.; De Groot, Anton C.; Slegers, Tanja P. A. M.; De Jong, Marcel C. J. M.; Pas, Hendri H.

    2009-01-01

    Diagnosis of pure ocular mucous membrane pemphigoid (OMMP) is based on the demonstration of linear depositions of IgG, IgA, C3 or combinations thereof alongside the epithelial basal membrane zone with direct immunofluorescence (DIF) or immunoperoxidase ( DIP) analysis of a freeze biopsy of the

  1. 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...

  2. Structure of microcin B-like compounds produced by Pseudomonas syringae and species specificity of their antibacterial action.

    Science.gov (United States)

    Metelev, Mikhail; Serebryakova, Marina; Ghilarov, Dmitry; Zhao, Youfu; Severinov, Konstantin

    2013-09-01

    Escherichia coli microcin B (Ec-McB) is a posttranslationally modified antibacterial peptide containing multiple oxazole and thiazole heterocycles and targeting the DNA gyrase. We have found operons homologous to the Ec-McB biosynthesis-immunity operon mcb in recently sequenced genomes of several pathovars of the plant pathogen Pseudomonas syringae, and we produced two variants of P. syringae microcin B (Ps-McB) in E. coli by heterologous expression. Like Ec-McB, both versions of Ps-McB target the DNA gyrase, but unlike Ec-McB, they are active against various species of the Pseudomonas genus, including human pathogen P. aeruginosa. Through analysis of Ec-McB/Ps-McB chimeras, we demonstrate that three centrally located unmodified amino acids of Ps-McB are sufficient to determine activity against Pseudomonas, likely by allowing specific recognition by a transport system that remains to be identified. The results open the way for construction of McB-based antibacterial molecules with extended spectra of biological activity.

  3. Molecular biology of deregulated gene expression in transformed C3H/10T1/2 mouse embryo cell lines induced by specific insoluble carcinogenic nickel compounds.

    Science.gov (United States)

    Landolph, Joseph R; Verma, Anuradha; Ramnath, Jamuna; Clemens, Farrah

    2002-10-01

    In the past, exposure of workers to mixtures of soluble and insoluble nickel compounds by inhalation during nickel refining correlated with increased incidences of lung and nasal cancers. Insoluble nickel subsulfide and nickel oxide (NiO) are carcinogenic in animals by inhalation; soluble nickel sulfate is not. Particles of insoluble nickel compounds were phagocytized by C3H/10T1/2 mouse embryo cells and induced morphological transformation in these cells with the following order of potency: NiO (black) > NiO (green) > nickel subsulfide. Foci induced by black/green NiO and nickel monosulfide developed into anchorage-independent transformed cell lines. Random arbitrarily primed-polymerase chain reaction mRNA differential display showed that nine c-DNA fragments are differentially expressed between nontransformed and nickel compound-transformed 10T1/2 cell lines in 6% of total mRNA; 130 genes would be differentially expressed in 100% of the mRNA. Fragment R3-2 was a sequence in the mouse calnexin gene, fragment R3-1 a portion of the Wdr1 gene, and fragment R2-4 a portion of the ect-2 protooncogene. These three genes were overexpressed in transformed cell lines. Fragment R1-2 was 90% homologous to a fragment of the DRIP/TRAP-80 (vitamin D receptor interacting protein/thyroid hormone receptor-activating protein 80) genes and was expressed in nontransformed but not in nickel-transformed cell lines. Specific insoluble carcinogenic nickel compounds are phagocytized into 10T1/2 cells and likely generate oxygen radicals, which would cause mutations in protooncogenes, and chromosome breakage, and mutations in tumor suppressor genes, inactivating them. These compounds also induce methylation of promoters of tumor suppressor genes, inactivating them. This could lead to permanent overexpresssion of the ect-2, calnexin, and Wdr1 genes and suppression of expression of the DRIP/TRAP-80 gene that we observed, which likely contribute to induction and maintenance of transformed

  4. Cytokeratin patterns in corneal, limbal, and conjunctival epithelium. An immunofluorescence study with PKK-1, 8.12, 8.60, and 4.62 anticytokeratin antibodies.

    Science.gov (United States)

    Ryder, M I; Weinreb, R N

    1990-11-01

    The authors examined immunofluorescently the specific cytokeratin staining patterns of corneal, limbal, and conjunctival epithelium with PKK-1, 8.12, 4.62, and 8.60 monoclonal anticytokeratin antibodies. Observations were made on unfixed frozen postmortem human tissue. The PKK-1 antibody stain was observed in all layers of corneal epithelium but only in suprabasal layers of limbal and conjunctival epithelium. By contrast, the 8.12 antibody stain was observed only in the superficial layer of corneal epithelium but through all layers of limbal and conjunctival epithelium. The 4.60 antibody stain was seen in focal areas of corneal and limbal epithelium and through all layers of conjunctival epithelium. The 8.60 antibody stain was not present in the three epithelia. These immunofluorescence studies showed unique cytokeratin patterns among layers in corneal, limbal, and conjunctival epithelium.

  5. Rapid determination of myosin heavy chain expression in rat, mouse, and human skeletal muscle using multicolor immunofluorescence analysis.

    Directory of Open Access Journals (Sweden)

    Darin Bloemberg

    Full Text Available Skeletal muscle is a heterogeneous tissue comprised of fibers with different morphological, functional, and metabolic properties. Different muscles contain varying proportions of fiber types; therefore, accurate identification is important. A number of histochemical methods are used to determine muscle fiber type; however, these techniques have several disadvantages. Immunofluorescence analysis is a sensitive method that allows for simultaneous evaluation of multiple MHC isoforms on a large number of fibers on a single cross-section, and offers a more precise means of identifying fiber types. In this investigation we characterized pure and hybrid fiber type distribution in 10 rat and 10 mouse skeletal muscles, as well as human vastus lateralis (VL using multicolor immunofluorescence analysis. In addition, we determined fiber type-specific cross-sectional area (CSA, succinate dehydrogenase (SDH activity, and α-glycerophosphate dehydrogenase (GPD activity. Using this procedure we were able to easily identify pure and hybrid fiber populations in rat, mouse, and human muscle. Hybrid fibers were identified in all species and made up a significant portion of the total population in some rat and mouse muscles. For example, rat mixed gastrocnemius (MG contained 12.2% hybrid fibers whereas mouse white tibialis anterior (WTA contained 12.1% hybrid fibers. Collectively, we outline a simple and time-efficient method for determining MHC expression in skeletal muscle of multiple species. In addition, we provide a useful resource of the pure and hybrid fiber type distribution, fiber CSA, and relative fiber type-specific SDH and GPD activity in a number of rat and mouse muscles.

  6. Congener-specific metabolism and sequestration of dioxin-like compounds by cytochrome P450 1A induced in the liver of crows from Tokyo, Japan

    Energy Technology Data Exchange (ETDEWEB)

    Watanabe, M.; Iwata, H.; Tanabe, S. [Ehime Univ., Matsuyama (Japan); Yoneda, K.; Hashimoto, T. [Japan Wildlife Research Center, Tokyo (Japan)

    2004-09-15

    Jungle crow (JC; Corvus macrorhynchos) is a useful bioindicator for monitoring contaminants in urban areas, because this species is residential, occupies a same habitat as human, and feeds variety of foods including domestic waste and garbage. Therefore, JCs may accumulate environmental contaminants such as polychlorinated dibenzo-p-dioxins (PCDDs), dibenzofurans (PCDFs) and coplanar polychlorinated biphenyls (Co-PCBs), which are released by human activities. Induction of cytochrome P450 (CYP) 1A is a responsive mechanism elicited by exposure to dioxinlike compounds including PCDDs/DFs and Co-PCBs. Toxicokinetic behavior of dioxin-like compounds in organisms is controlled by excretion, metabolism and absorption. These processes are, at least partly, dependent on CYP1A expression in addition to chemical structure and number of chlorine substitution of each congener. Low chlorinated congeners such as 2378-T{sub 4}CDD, 2378- T{sub 4}CDF, 12378-P{sub 5}CDD and 33'44'-PCB were easily metabolized by CYP1A1/2 in rat liver microsomes. PCDDs/DFs accumulate in hepatic tissue to a greater extent than adipose tissue in rats and mice. Recent study using transgenic CYP1A2 knockout mice demonstrated that CYP1A2 is responsible for the sequestration of 2378-T{sub 4}CDD and 23478-P{sub 5}CDF in hepatic tissue. Therefore, CYP1A is considered as a key factor responsible for toxicokinetics of dioxin-like compounds. However, there's no comprehensive data on the contribution of CYP1A to the toxicokinetics of dioxin-like congeners in wild populations. In this study, we investigated contamination levels of PCDDs/DFs and Co-PCBs in liver and breast muscle of JCs from Tokyo, Japan, and interactions of dioxin-like congeners with hepatic CYP to elucidate congener-specific toxicokinetics related to CYP expression in JC.

  7. C, Cl and H compound-specific isotope analysis to assess natural versus Fe(0) barrier-induced degradation of chlorinated ethenes at a contaminated site

    Energy Technology Data Exchange (ETDEWEB)

    Audí-Miró, Carme, E-mail: carmeaudi@ub.edu [Grup de Mineralogia Aplicada i Medi Ambient, Departament de Cristal.lografia, Mineralogia i Dipòsits Minerals, Facultat de Geologia, Universitat de Barcelona (UB), Martí Franquès s/n, 08028, Barcelona (Spain); Cretnik, Stefan [Institute of Groundwater Ecology, Helmholtz Zentrum München-National Research Center for Environmental Health, Ingolstädter Landstrasse 1, D-85764 Neuherberg (Germany); Torrentó, Clara; Rosell, Mònica [Grup de Mineralogia Aplicada i Medi Ambient, Departament de Cristal.lografia, Mineralogia i Dipòsits Minerals, Facultat de Geologia, Universitat de Barcelona (UB), Martí Franquès s/n, 08028, Barcelona (Spain); Shouakar-Stash, Orfan [Department of Earth & Environmental Sciences, 200 University Ave. W, N2L 3G1 Waterloo, Ontario (Canada); Otero, Neus [Grup de Mineralogia Aplicada i Medi Ambient, Departament de Cristal.lografia, Mineralogia i Dipòsits Minerals, Facultat de Geologia, Universitat de Barcelona (UB), Martí Franquès s/n, 08028, Barcelona (Spain); Palau, Jordi [Université de Neuchâtel, CHYN - Centre d' Hydrogéologie, Rue Emile-Argand 11, CH-2000 Neuchâtel (Switzerland); and others

    2015-12-15

    Highlights: • {sup 13}C to evaluate natural chlorinated ethenes biodegradation. • {sup 13}C to evaluate the efficiency of a zero-valent iron-permeable reactive barrier. • {sup 13}C-{sup 37}Cl to discriminate biotic from abiotic degradation of cis-dichloroethene. • {sup 13}C-{sup 37}Cl-{sup 2}H of cis-DCE and TCE to elucidate different contaminant sources. - Abstract: Compound-specific isotopic analysis of multiple elements (C, Cl, H) was tested to better assess the effect of a zero-valent iron-permeable reactive barrier (ZVI-PRB) installation at a site contaminated with tetrachloroethene (PCE) and trichloroethene (TCE). The focus was on (1) using {sup 13}C to evaluate natural chlorinated ethene biodegradation and the ZVI-PRB efficiency; (2) using dual element {sup 13}C-{sup 37}Cl isotopic analysis to distinguish biotic from abiotic degradation of cis-dichloroethene (cis-DCE); and (3) using {sup 13}C-{sup 37}Cl-{sup 2}H isotopic analysis of cis-DCE and TCE to elucidate different contaminant sources. Both biodegradation and degradation by ZVI-PRB were indicated by the metabolites that were detected and the {sup 13}C data, with a quantitative estimate of the ZVI-PRB efficiency of less than 10% for PCE. Dual element {sup 13}C-{sup 37}Cl isotopic plots confirmed that biodegradation was the main process at the site including the ZVI-PRB area. Based on the carbon isotope data, approximately 45% and 71% of PCE and TCE, respectively, were estimated to be removed by biodegradation. {sup 2}H combined with {sup 13}C and {sup 37}Cl seems to have identified two discrete sources contributing to the contaminant plume, indicating the potential of δ{sup 2}H to discriminate whether a compound is of industrial origin, or whether a compound is formed as a daughter product during degradation.

  8. Nitrite curing of chicken, pork, and beef inhibits oxidation but does not affect N-nitroso compound (NOC)-specific DNA adduct formation during in vitro digestion.

    Science.gov (United States)

    Van Hecke, Thomas; Vanden Bussche, Julie; Vanhaecke, Lynn; Vossen, Els; Van Camp, John; De Smet, Stefaan

    2014-02-26

    Uncured and nitrite-cured chicken, pork, and beef were used as low, medium, and high sources of heme-Fe, respectively, and exposed to an in vitro digestion model simulating the mouth, stomach, duodenum, and colon. With increasing content of iron compounds, up to 25-fold higher concentrations of the toxic lipid oxidation products malondialdehyde, 4-hydroxy-2-nonenal, and other volatile aldehydes were formed during digestion, together with increased protein carbonyl compounds as measurement of protein oxidation. Nitrite curing of all meats lowered lipid and protein oxidation to the level of oxidation in uncured chicken. Strongly depending on the individual fecal inoculum, colonic digestion of beef resulted in significantly higher concentrations of the NOC-specific DNA adduct O(6)-carboxymethyl-guanine compared to chicken and pork, whereas nitrite curing had no significant effect. This study confirms previously reported evidence that heme-Fe is involved in the epidemiological association between red meat consumption and colorectal cancer, but questions the role of nitrite curing in this association.

  9. Tracing fresh assimilates through Larix decidua exposed to elevated CO₂ and soil warming at the alpine treeline using compound-specific stable isotope analysis.

    Science.gov (United States)

    Streit, Kathrin; Rinne, Katja T; Hagedorn, Frank; Dawes, Melissa A; Saurer, Matthias; Hoch, Günter; Werner, Roland A; Buchmann, Nina; Siegwolf, Rolf T W

    2013-02-01

    How will carbon source-sink relations of 35-yr-old larch trees (Larix decidua) at the alpine treeline respond to changes in atmospheric CO(2) and climate? We evaluated the effects of previously elevated CO(2) concentrations (9 yr, 580 ppm, ended the previous season) and ongoing soil warming (4 yr, + 4°C). Larch branches were pulse labeled (50 at% (13)CO(2)) in July 2010 to trace fresh assimilates through tissues (buds, needles, bark and wood) and non-structural carbon compounds (NCC; starch, lipids, individual sugars) using compound-specific isotope analysis. Nine years of elevated CO(2) did not lead to increased NCC concentrations, nor did soil warming increase NCC transfer velocities. By contrast, we found slower transfer velocities and higher NCC concentrations than reported in the literature for lowland larch. As a result of low dilution with older carbon, sucrose and glucose showed the highest maximum (13)C labels, whereas labels were lower for starch, lipids and pinitol. Label residence times in needles were shorter for sucrose and starch (c. 2 d) than for glucose (c. 6 d). Although our treatments showed no persistent effect on larch carbon relations, low temperature at high altitudes clearly induced a limitation of sink activities (growth, respiration, root exudation), expressed in slower carbon transfer and higher NCC concentrations. © 2012 The Authors. New Phytologist © 2012 New Phytologist Trust.

  10. Identification of animal fats via compound specific δ13C values of individual fatty acids: assessments of results for reference fats and lipid extracts of archaeological pottery vessels

    Directory of Open Access Journals (Sweden)

    Richard P. Evershed

    2002-12-01

    Full Text Available The possibility of obtaining molecular information from lipid residues associated with archaeological pottery has dramatically increased the potential for deriving new information on the use of ancient vessels and the commodities processed therein. Motivated by the high proportion of the archaeological potsherds that have been shown to contain animal fats, a new approach invol- ving compound specific stable isotope analysis of remnant fats has been developed to retrieve infor- mation which will allow new insights into animal exploitation, dietary preferences and vessel use amongst prehistoric peoples. The new approach uses the δ13C values of the major saturated fatty acid (C16:0 and C18:0 determined by gas chromatography-combustion-isotope ratio mass spectrometry (GC–C–IRMS to characterise the origins of animal fat recovered from archaeological pottery.

  11. Fingerprinting TCE Sources and Natural Attenuation Capacity of a Sedimentary Bedrock Aquifer using Compound Specific Isotope Analysis - A Case Study in the City of Guelph, Ontario.

    Science.gov (United States)

    Lima, P. L.; Aravena, R.; Parker, B. L.; Cherry, J. A.

    2009-05-01

    Chlorinated volatile organic compounds were detected in an industrial facility (site) in the upper part of the underlying dolomite bedrock aquifer. The primary chlorinated solvent detected at the site is trichloroethene (TCE), and associated daughter products, such as cis-1,2- dichloroethene (cis-1,2-DCE) and vinyl chloride (VC), are also present. Several conventional monitoring wells installed at the site were recently sampled for VOCs analysis, inorganic sensitive REDOX parameters and Compound Specific Isotope Analyses (CSIA) of 13C and 37Cl in order to evaluate the natural attenuation capacity of the local upper bedrock aquifer. A municipal well was also included in this study. The municipal well is located approximately 500 meters southwest from the investigated site and consists of anuncased bedrock well completed to 102 meters below ground surface (mbgs). Chlorinated volatile organic compounds were detected in this well during the evaluation of municipal groundwater resources performed in the early 1990s by the City of Guelph. Neither the City nor the Ministry of the Environment of the Province of Ontario (MOE) have been able to satisfactorily determine the origin of these compounds. Groundwater for VOCs analysis and CSIA were therefore collected in the municipal well in order to evaluate the relationship between the TCE detected in the municipal well and the TCE plume detected in the nearby industrial facility. The inorganic data indicated that the local upper aquifer is under sulfate-reducing conditions in some areas where typical VOC's daughter products and highly enriched δ13C and δ 37Cl values of TCE and cis-1,2-DCE were observed. These findings indicated that microbial reductive dechlorination of TCE is occurring in some areas of the local aquifer. The δ 13C values of TCE obtained in groundwater from the conventional wells were approximately 10 ‰ more depleted than δ 13C values obtained for the deep municipal production well. These data indicate

  12. Sources of variability in fatty acid (FA) biomarkers in the application of compound-specific stable isotopes (CSSIs) to soil and sediment fingerprinting and tracing: A review

    Energy Technology Data Exchange (ETDEWEB)

    Reiffarth, D.G., E-mail: Dominic.Reiffarth@unbc.ca [Natural Resources and Environmental Studies Program, University of Northern British Columbia, 3333 University Way, Prince George, BC V2N 4Z9 (Canada); Petticrew, E.L., E-mail: Ellen.Petticrew@unbc.ca [Geography Program and Quesnel River Research Centre, University of Northern British Columbia, 3333 University Way, Prince George, BC V2N 4Z9 (Canada); Owens, P.N., E-mail: Philip.Owens@unbc.ca [Environmental Science Program and Quesnel River Research Centre, University of Northern British Columbia, 3333 University Way, Prince George, BC, V2N 4Z9 (Canada); Lobb, D.A., E-mail: David.Lobb@umanitoba.ca [Watershed Systems Research Program, University of Manitoba, 13 Freedman Crescent, Winnipeg, MB R3T 2N2 (Canada)

    2016-09-15

    Determining soil redistribution and sediment budgets in watersheds is often challenging. One of the methods for making such determinations employs soil and sediment fingerprinting techniques, using sediment properties such as geochemistry, fallout radionuclides, and mineral magnetism. These methods greatly improve the estimation of erosion and deposition within a watershed, but are limited when determining land use-based soil and sediment movement. Recently, compound-specific stable isotopes (CSSIs), which employ fatty acids naturally occurring in the vegetative cover of soils, offer the possibility of refining fingerprinting techniques based on land use, complementing other methods that are currently in use. The CSSI method has been met with some success; however, challenges still remain with respect to scale and resolution due to a potentially large degree of biological, environmental and analytical uncertainty. By better understanding the source of tracers used in CSSI work and the inherent biochemical variability in those tracers, improvement in sample design and tracer selection is possible. Furthermore, an understanding of environmental and analytical factors affecting the CSSI signal will lead to refinement of the approach and the ability to generate more robust data. This review focuses on sources of biological, environmental and analytical variability in applying CSSI to soil and sediment fingerprinting, and presents recommendations based on past work and current research in this area for improving the CSSI technique. A recommendation, based on current information available in the literature, is to use very-long chain saturated fatty acids and to avoid the use of the ubiquitous saturated fatty acids, C{sub 16} and C{sub 18}. - Highlights: • Compound-specific stable isotopes (CSSIs) of carbon may be used as soil tracers. • The variables affecting CSSI data are: biological, environmental and analytical. • Understanding sources of variability will lead

  13. Reconstructing hydroclimatic variations using compound-specific hydrogen isotope analysis of biomarkers from a maar lake in the Central Highlands, Vietnam

    Science.gov (United States)

    Doiron, Kelsey; Stevens, Lora; Sauer, Peter

    2017-04-01

    Monsoonal variation in Southeast Asia affects a significant portion of the global population, but knowledge regarding response of the monsoon system to changing boundary conditions is limited. The paleoclimatic tool of compound-specific isotope analysis(CSIA) provides the ability to reconstruct past precipitation using a diverse set of biomarkers preserved in the sedimentary record. Limited proxies in tropical southeast Asia and difficult site access have led to a deficit in paleoclimate records. Ia M'He (14˚ 10'45" N, 107˚ 52' E) is a shallow volcanic crater (maar) lake, approximately 57 ha, located in the Central Highlands of Vietnam. Precipitation in the Central Highlands is sensitive to reorganizations of major climatic features, such as the migration of the ITCZ and the coupled Indo-Asian monsoon, ENSO and related shifts in the Pacific Walker Circulation and typhoon frequency. To examine this complex behavior, this pilot study aims to provide a 500-year record of effective moisture inferred from CSIA of hydrogen isotopes on biomarkers. Carbon/nitrogen ratios and carbon isotope ratios indicate that bulk organic matter is a combination of algae and C3 vegetation, offering the potential to use compound-specific hydrogen isotopes of aquatic and terrestrial organic matter in tandem. Preliminary analysis of the core shows dominant alkane chain lengths of C27 and C29, associated with terrestrial plant leaf waxes. The hydrogen isotope ratios of the plant wax components provide a proxy for paleo precipitation in a region where rainfall and droughts heavily influence population dynamics and create social discord. The CSIA record is expected to correlate with records from northern Vietnam, the South China Sea and Indonesia, with greater precipitation during the Little Ice Age. The degree to which evaporative modification of lake water (i.e., seasonal drying) occurs will be estimated by comparing the terrestrial CSIA values indicative of meteoric water with aquatic CSIA

  14. Compound-specific 15N stable isotope probing of N assimilation by the soil microbial biomass: a new methodological paradigm in soil N cycling

    Science.gov (United States)

    Charteris, A. F.; Knowles, T. D. J.; Michaelides, K.; Evershed, R. P.

    2015-10-01

    A compound-specific nitrogen-15 stable isotope probing (15N-SIP) technique is described which allows investigation of the fate of inorganic- or organic-N amendments to soils. The technique uses gas chromatography-combustion-isotope ratio mass spectrometry (GC-C-IRMS) to determine the δ15N values of individual amino acids (AAs; determined as N-acetyl, O-isopropyl derivatives) as proxies of biomass protein production. The δ15N values are used together with AA concentrations to quantify N assimilation of 15N-labelled substrates by the soil microbial biomass. The utility of the approach is demonstrated through incubation experiments using inorganic 15N-labelled substrates ammonium (15NH4+) and nitrate (15NO3-) and an organic 15N-labelled substrate, glutamic acid (15N-Glu). Assimilation of all the applied substrates was undetectable based on bulk soil properties, i.e. % total N (% TN), bulk soil N isotope composition and AA concentrations, all of which remained relatively constant throughout the incubation experiments. In contrast, compound-specific AA δ15N values were highly sensitive to N assimilation, providing qualitative and quantitative insights into the cycling and fate of the applied 15N-labelled substrates. The utility of this 15N-AA-SIP technique is considered in relation to other currently available methods for investigating the microbially-mediated assimilation of nitrogenous substrates into the soil organic N pool. This approach will be generally applicable to the study of N cycling in any soil, or indeed, in any complex ecosystem.

  15. Babesial antibody dynamics after cattle immunisation with live vaccines, measured with an indirect immunofluorescence test.

    Science.gov (United States)

    Guglielmone, A A; Lugaresi, C I; Volpogni, M M; Anziani, O S; Vanzini, V R

    1997-06-01

    The efficacy of vaccination of Argentinean cattle against babesiosis and anaplasmosis using live immunogens was tested to detect specific antibodies in samples obtained about 60 days after vaccination. Under these conditions a higher than expected proportion of cattle failed to show antibodies against Babesia bigemina. Therefore, a study was designed to evaluate if this failure was due to insensitivity of the routine test to detect antibodies to B. bigemina or to lack of infectivity of the live vaccine. Four groups (G) of cattle were each inoculated subcutaneously with 10 million Babesia bovis (vaccinal strain R1A), 10 million B. bigemina (vaccinal strain S1A) and 10 million Anaplasma centrale (strain M1). G1 and G2 consisted of ten Angus bulls 20-24 months old and ten Angus bulls 15-18 months old, respectively; G3 and G4 consisted of ten and 16 Holstein 1-month-old male calves, respectively. Blood samples were obtained on days 0, 10, 20, 30, 40, 50 and 60 after vaccination and the sera were analysed with an indirect immunofluorescent (IFA) test to detect antibodies to B. bovis (baseline dilution for a positive result 1:60) and B. bigemina (baseline dilution 1:120). Positive IFA titres were considered as evidence of the infectivity of the Babesia vaccinal strains contained in the vaccine. All Angus bulls were found positive to antibodies against both Babesia species, by day 20 (B. bovis) and day 30 (B. bigemina), whereas 10-25% of Holstein calves were negative throughout. The partial lack of vaccine infectivity in the calves was considered to be a consequence of innate resistance of young calves to Babesia. Antibody titres to B. bovis and B. bigemina declined by day 60 after vaccination. However, all cattle that were positive to B. bovis antibodies on day 50 were still positive to the IFA test 10 days later while 10%, 30% and 12% of cattle of G1, G2 and G3 that were positive to B. bigemina antibodies on day 50 after vaccination were found negative to the IFA test

  16. Targeting Prostate-Specific Membrane Antigen (PSMA) with F-18-Labeled Compounds: the Influence of Prosthetic Groups on Tumor Uptake and Clearance Profile.

    Science.gov (United States)

    Bouvet, Vincent; Wuest, Melinda; Bailey, Justin J; Bergman, Cody; Janzen, Nancy; Valliant, John F; Wuest, Frank

    2017-12-01

    Prostate-specific membrane antigen (PSMA) is an important biomarker expressed in the majority of prostate cancers. The favorable positron emission tomography (PET) imaging profile of the PSMA imaging agent 2-(3-(1-carboxy-5-[(6-[(18)F]fluoro-pyridine-3-carbonyl)-amino]-pentyl)-ureido)-pentane-dioic acid [(18)F]DCFPyL in preclinical prostate cancer models and in prostate cancer patients stimulated the development and validation of other fluorine-containing PSMA inhibitors to further enhance pharmacokinetics and simplify production methods. Here, we describe the synthesis and radiopharmacological evaluation of various F-18-labeled PSMA inhibitors which were prepared through different prosthetic group chemistry strategies. Prosthetic groups N-succinimidyl-4-[(18)F]fluorobenzoate ([(18)F]SFB), 4-[(18)F]fluorobenzaldehyde, and 2-deoxy-2-[(18)F]fluoro-D-glucose ([(18)F]FDG) were used for bioconjugation reactions to PSMA-binding lysine-urea-glutamate scaffold via acylation and oxime formation. All fluorine-containing PSMA inhibitors were tested for their PSMA inhibitory potency in an in vitro competitive binding assay in comparison to an established reference compound [(125)I]TAAG-PSMA. Tumor uptake and clearance profiles of three F-18-labeled PSMA inhibitors ([(18)F]4, [(18)F]7, and [(18)F]8) were studied with dynamic PET imaging using LNCaP tumor-bearing mice. F-18-labeled PSMA inhibitors were synthesized in 32-69 % radiochemical yields using (1) acylation reaction at the primary amino group of the lysine residue with [(18)F]SFB and (2) oxime formation with 4-[(18)F]fluorobenzaldehyde and [(18)F]FDG using the respective aminooxy-functionalized lysine residue. Compound 7 displayed an IC50 value of 6 nM reflecting very high affinity for PSMA. Compounds 4 and 8 showed IC50 values of 13 and 62 nM, respectively. The IC50 value of reference compound DCFPyL was 13 nM. Dynamic PET imaging revealed the following SUV60min for radiotracer uptake in PSMA(+) LNCaP tumors: 0

  17. Microwave oven-based technique for immunofluorescent staining of paraffin-embedded tissues.

    Science.gov (United States)

    Long, Delwin J; Buggs, Colleen

    2008-02-01

    Immunohistochemical analysis of formalin-fixed paraffin-embedded tissues can be challenging due to potential modifications of protein structure by exposure to formalin. Heat-induced antigen retrieval techniques can reverse reactions between formalin and proteins that block antibody recognition. Interactions between antibodies and antigens are further enhanced by microwave irradiation, which has simplified immunohistochemical staining protocols. In this report, we modify a technique for antigen retrieval and immunofluorescent staining of formalin-fixed paraffin-embedded tissues by showing that it works well with several antibodies and buffers. This microwave-assisted method for antigen retrieval and immunofluorescent staining eliminates the need for blocking reagents and extended washes, which greatly simplifies the protocol allowing one to complete the analysis in less than 3 h.

  18. Heterogeneity of antimitochondrial antibodies with the M2-M4 pattern by immunofluorescence as assessed by Western immunoblotting and enzyme linked immunosorbent assay.

    Science.gov (United States)

    Fusconi, M; Ghadiminejad, I; Bianchi, F B; Baum, H; Bottazzo, G F; Pisi, E

    1988-01-01

    Seventy seven sera with antimitochondrial antibody exhibiting the M2-M4 pattern in immunofluorescence (56 from primary biliary cirrhosis (PBC), 21 from non-primary biliary cirrhosis patients) were studied by the combined use of Western immunoblotting with beef heart mitochondria and an enzyme linked immunosorbent assay (ELISA) with beef heart submitochondrial particles. Forty seven sera (10 without autoantibodies and 37 with different auto-antibodies) were included as controls. By immunoblotting, seven mitochondrial peptides reacting with antimitochondrial antibody positive sera were detected. These were of molecular weight 74 kD, 58 kD, 55 kD, 52 kD, 51 kD, 46 kD, and 43 kD. All primary biliary cirrhosis sera and 71% of antimitochondrial antibody-positive non-primary biliary cirrhosis sera reacted with one or more of these peptides, while none of the 47 antimitochondrial antibody negative sera reacted in immunoblotting. The 74 kD band was the most frequently detected (84% of primary biliary cirrhosis and 57% of non-primary biliary cirrhosis cases). All the primary biliary cirrhosis sera which failed to react with this peptide, showed a positive reaction with that of molecular weight 52 kD. 67/77 (87%) immunofluorescence antimitochondrial antibody positive sera reacted in the ELISA test (93% of primary biliary cirrhosis and 71% of non-primary biliary cirrhosis cases). All the 47 immunofluorescence antimitochondrial antibody negative sera were confirmed negative by ELISA. The ELISA values correlated with the immunofluorescence titres (p less than 0.05). By comparison of the results obtained by these two techniques, it emerged that the ELISA test (using our preparation of submitochondrial particles) was not able to detect the antibody directed against the mitochondrial peptide of 52 kD, which thus seems to be different from the other specificities. Images Fig. 1 PMID:2453397

  19. Triple immunofluorescence labeling of atherosclerotic plaque components in apoE/LDLR -/- mice.

    OpenAIRE

    Stefan Chłopicki; Andrzej Radziszewski; Grzegorz J Lis; Lukasz Mateuszuk; Jacek Jawień; Mariusz Gajda; Jan A Litwin

    2008-01-01

    This paper presents a simple and reliable method of triple immunofluorescence staining that allows simultaneous detection of various cell types present in atherosclerotic plaque of apolipoprotein E and LDL receptor-double knockout (apoE/LDLR -/-) mice. We used combined direct and indirect procedures applying commercially available primary antibodies raised in different species to detect smooth muscle cells (Cy3-conjugated mouse anti-smooth muscle actin, SMA), macrophages (rat anti-CD68) and T...

  20. Evaluation of three immunofluorescence assays for culture confirmation and typing of herpes simplex virus.

    OpenAIRE

    Lipson, S M; Schutzbank, T E; Szabo, K

    1987-01-01

    Three pairs of monoclonal antibodies, supplied in kits by Electro-Nucleonics, Inc. (ENI), The Syva Co., and Kallestad Laboratories, Inc. (KL), were evaluated for the laboratory confirmation and typing of herpes simplex virus (HSV). Of 108 coded HSV slide preparation, run in parallel with each monoclonal-antibody set, 103 were equivalent by the immunofluorescence assays. Among the five discordant isolates, three (2.8%) did not type with the KL monoclonal antibodies and two (1.9%) false-positiv...

  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. In vitro evaluation of a specific radiochemical compound based on 99mTc-labeled DARPinG3 for radionuclide imaging of tumors overexpressing Her-2/neu

    Science.gov (United States)

    Bragina, O.; Larkina, M.; Stasyuk, E.; Chernov, V.; Zelchan, R.; Medvedeva, A.; Sinilkin, I.; Yusubov, M.; Skuridin, V.; Deyev, S.; Buldakov, M.

    2017-09-01

    It is still necessary to search for new informative diagnostic methods to detect malignant tumors with overexpression of Her-2/neu, which are characterized by the aggressive course of the disease, rapid rate of tumor growth and low rates of relapse-free and overall survival. In recent years, the radioisotope techniques for detection of specific tumor targets have been developing actively. Purpose: to develop a chemically stable radiochemical compound for the targeted imaging of cells overexpressing Her-2/neu. Material and methods: The study was performed using 2 cell lines. The human breast adenocarcinoma HER2-overexpressing cell line BT-474 was chosen to detect specific binding. As a control, HER2-negative human breast adenocarcinoma MCF-7 was used. The human breast adenocarcinoma BT-474 and MCF-7 cell lines were seeded in chamber-slides at the density of 35,000 cells/ml in trypsin-EDTA (PanEco) medium and grown overnight at 37°C. After that both cell lines were washed with Phosphate buffered saline (PBS) and distributed into test tubes to 1 ml (5 millions cells in each). After adding 100 µl (70 MBq) studied complex of 99mTc-DPAH- DARPinG3 was incubated for 40 min at +4°C. Washing was performed three times with buffer PBS and 5% Bovine Serum Albumin (BSA). The characteristics of the binding specificity of the test set with the HER-2/neu receptor were determined by direct radiometric and planar scintigraphy. Nonparametric Mann-Whitney test was used to assess the differences in the quantitative characteristics between groups. Results: The output of the labeled complex was more than 91%, with a radiochemical purity of more than 94%. When carrying out a visual scintigraphic assessment much greater intensity accumulation of radiotracer was observed in the studied cell culture surface receptor overexpressing Her-2/neu. The results of direct radiometric also showed higher accumulation of the radiopharmaceutical in the adenocarcinoma cell line BT-474 human breast

  3. Direct Tracking and Compound-Specific Stable Isotope Analysis Identifies Dispersal Routes and Cryptic Latitudinal Trends in the Winter Distribution of Two Antarctic Penguin Species

    Science.gov (United States)

    Polito, M. J.; Houghton, L. A.; Hinke, J.; Thorrold, S.

    2016-02-01

    Animal dispersal behaviors have both ecological and evolutionary significance. However, due to the logistical challenges of tracking animals in the Antarctic marine environment little is known about the winter dispersal and migration of Antarctic marine predators. We used a combination of direct tracking (Geolocation sensors; GLS) and compound-specific stable isotope analysis of carbon in essential amino acids (CSIA-AA) to describe the winter distribution of two wide-ranging marine predators, the Adélie penguin (Pygoscelis adeliae) and chinstrap penguin (P. antarctica) along the Antarctic Peninsula and Scotia Arc. GLS and CSIA-AA identified two general migration strategies, with animals overwintering to the east or west of the Antarctic Peninsula, with CSIA-AA also being able to discriminate between ice-free vs. pack-ice habitats. In addition, CSIA-AA provided the ability to assign non GLS-tracked chinstrap penguins from multiple breeding sites across the Scotia Arc to specific over-wintering areas. This provided a first ever, regional-scale analyses of the winter movements and distributions of the species and identified a cryptic latitudinal trend that would have been otherwise undetectable. Breeding colonies farther north were more likely to have individuals dispersing eastwards relative to colonies further south where most individual dispersed westwards, possibly due to a combination of proximity and competition avoidance. Finally we highlight how CSIA-AA approaches can be applied to other studies of the distribution and ecology of marine top predators.

  4. Highly Active and Specific Tyrosine Ammonia-Lyases from Diverse Origins Enable Enhanced Production of Aromatic Compounds in Bacteria and Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    Jendresen, Christian Bille; Stahlhut, Steen Gustav; Li, Mingji

    2015-01-01

    Phenylalanine and tyrosine ammonia-lyases form cinnamic acid and p-coumaric acid, which are precursors of a wide range of aromatic compounds of biotechnological interest. Lack of highly active and specific tyrosine ammonia-lyases has previously been a limitation in metabolic engineering approache...... were also efficient in Saccharomyces cerevisiae, where p-coumaric acid accumulation was improved 5-fold over that in strains expressing previously characterized tyrosine ammonia-lyases....... Dictyostelium discoideum, and a phenylalanine ammonia-lyase from the moss Physcomitrella patens (producing 230 μM cinnamic acid per unit of optical density at 600 nm [OD600]) in the medium using Escherichia coli as the heterologous host). Novel tyrosine ammonia-lyases having higher reported substrate...... specificity than previously characterized enzymes were also identified. Enzymes from Herpetosiphon aurantiacus and Flavobacterium johnsoniae resulted in high production of p-coumaric acid in Escherichia coli (producing 440 μM p-coumaric acid OD600 unit−1 in the medium) and in Lactococcus lactis. The enzymes...

  5. Compound-Specific Carbon, Nitrogen, and Hydrogen Isotopic Ratios for Amino Acids in CM and CR Chondrites and their use in Evaluating Potential Formation Pathways

    Science.gov (United States)

    Elsila, Jamie E.; Charnley, Steven B.; Burton, Aaron S.; Glavin, Daniel P.; Dworkin, Jason P.

    2012-01-01

    Stable hydrogen, carbon, and nitrogen isotopic ratios (oD, 013C, and olSN) of organic compounds can revcal information about their origin and formation pathways. Several formation mechanisms and environments have been postulated for the amino acids detected in carbonaceous chondrites. As each proposed mechanism utilizes different precursor molecules, the isotopic signatures of the resulting amino acids may indicate the most likely of these pathways. We have applied gas chromatography with mass spectrometry and combustion isotope ratio mass spectrometry to measure the compound-specific C, N, and H stable isotopic ratios of amino acids from seven CM and CR carbonaceous chondrites: CM1I2 Allan Hills (ALH) 83100, CM2 Murchison, CM2 Lewis Cliff (LEW) 90500, CM2 Lonewolf Nunataks (LON) 94101, CRZ Graves Nunataks (GRA) 95229, CRZ Elephant Moraine (EET) 92042, and CR3 Queen Alexandra Range (QUE) 99177. We compare the isotopic compositions of amino acids in these meteorites with predictions of expected isotopic enrichments from potential formation pathways. We observe trends of decreasing ODC and increasing oD with increasing carbon number in the aH, (l-NH2 amino acids that correspond to predictions made for formation via Streckercyanohydrin synthesis. We also observe light ODC signatures for -alanine, which may indicate either formation via Michael addition or via a pathway that forms primarily small, straight-chain, amine-terminal amino acids (n-ro-amino acids). Higher deuterium enrichments are observed in amethyl amino acids, indicating formation of these amino acids or their precursors in cold interstellar or nebular environments. Finally, individual amino acids are more enriched in deuterium in CR chondrites than CM chondrites, reflecting different parent-body chemistry.

  6. 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

  7. Symptomatology and diagnosis in connective tissue disease. Antibodies to extractable ribonucleoprotein in 123 patients reacting with cell nuclei in the immunofluorescence test.

    Science.gov (United States)

    Jonsson, J

    1976-01-01

    Sera with an antinuclear immunofluorescence titre of 1/000 were taken consecutively from the diagnostic routine flow and examined for agglutinating antibodies against desoxyribonucleic acid (DNA) and extractable nuclear antigens (ENA). Passive haemagglutination tests with antigen-coated tanned erythrocytes were used and the specificity of the reactions was corroborated by testing against enzyme-treated cells. After the exclusion of the DNA-reacting 15%, three major groups and one minor could be distinguished on a serological basis. The largest group (41%) contained cases with a speckled immunofluorescence pattern and a RNase-trypsin sensitive agglutination reaction with ENA coated cells (sRNP). Nearly all cases of mixed connective tissue disease and scleroderma fell into this group which also contained 44% of the SLE cases. Symptomatically the group was characterized by remarkably high incidences of Raynaud's syndrome and myositis. The major group next in size comprised cases with a homogeneous immunofluorescence pattern but no reaction against DNA or ENA. Half of the cases within this group had the diagnosis SLE; they also constituted 42% of all SLE cases. The only other diagnosis of significant frequency within the group was unspecified collagenosis (23%). The symptomatology of the group was rather uncharacteristic, with the exception of the low incidence of Raynaud's syndrome. The third major group comprised cases with a speckled immunofluorescence pattern but no agglutination reaction against ENA or DNA. This group had a very high incidence of rheumatoid factor and also the highest incidence of visceral lesions among the groups. Yet the group contained only a small proportion (14%) of the SLE cases and the rheumatoid arthritis cases were about equally shared between this and the first group. The most common diagnosis in the group was unspecified collagenosis (40%). A fourth, small but homogeneous group contained cases with a speckled immunofluorescence pattern

  8. Studies on colony specificity in the compound ascidian, Botryllus primigenus Oka. I. Initiation of "nonfusion" reaction with special reference to blood cells infiltration.

    Science.gov (United States)

    Taneda, Y; Watanabe, H

    1982-01-01

    When two incompatible colonies of the compound ascidian, Botryllus primigenus Oka come into contact, a specific rejection, called "nonfusion" reaction (NFR), always occurs in the contact area between them. At the beginning of the NFR, concentrations of cells were always observed near the ampullae tips. In the present study, it was revealed that these cells appear to be blood cells infiltrating through the ampullae and were identified as morula cells (vanadocytes). When India ink was injected into vessels prior to the occurrence of NFR, it leaked from the tips of the ampullae into the test matrix, suggesting an increase in the permeability of the ampullar epidermis. Experimental studies to elucidate the mechanism of NFR were carried out. Contact between a colony piece and a zooid-free ampullae fragment or overlapping of one colony to another, respectively, resulted in NFR only at one side of a pair. Furthermore, in stage 3 of NFR, when an opaque area was pushed with a pair of forceps and the blood in this area was transferred to healthy part of vessels, the stoppage of blood flow and subsequent contraction of vessels always took place in the latter. These observations suggest that ampullae penetration into the opposite colony is essential to NFR and that NFR is directly initiated in the ampullar lumen.

  9. Species-specific diversity of novel bacterial lineages and differential abundance of predicted pathways for toxic compound degradation in scorpion gut microbiota.

    Science.gov (United States)

    Bolaños, Luis M; Rosenblueth, Mónica; Castillo-Ramírez, Santiago; Figuier-Huttin, Gilles; Martínez-Romero, Esperanza

    2016-05-01

    Scorpions are considered 'living fossils' that have conserved ancestral anatomical features and have adapted to numerous habitats. However, their gut microbiota diversity has not been studied. Here, we characterized the gut microbiota of two scorpion species, Vaejovis smithi and Centruroides limpidus. Our results indicate that scorpion gut microbiota is species-specific and that food deprivation reduces bacterial diversity. 16S rRNA gene phylogenetic analysis revealed novel bacterial lineages showing a low level of sequence identity to any known bacteria. Furthermore, these novel bacterial lineages were each restricted to a different scorpion species. Additionally, our results of the predicted metagenomic profiles revealed a core set of pathways that were highly abundant in both species, and mostly related to amino acid, carbohydrate, vitamin and cofactor metabolism. Notably, the food-deprived V. smithi shotgun metagenome matched almost completely the metabolic features of the prediction. Finally, comparisons among predicted metagenomic profiles showed that toxic compound degradation pathways were more abundant in recently captured C. limpidus scorpions. This study gives a first insight into the scorpion gut microbiota and provides a reference for future studies on the gut microbiota from other arachnid species. © 2015 Society for Applied Microbiology and John Wiley & Sons Ltd.

  10. Characterization of toluene and ethylbenzene biodegradation under nitrate-, iron(III)- and manganese(IV)-reducing conditions by compound-specific isotope analysis.

    Science.gov (United States)

    Dorer, Conrad; Vogt, Carsten; Neu, Thomas R; Stryhanyuk, Hryhoriy; Richnow, Hans-Hermann

    2016-04-01

    Ethylbenzene and toluene degradation under nitrate-, Mn(IV)-, or Fe(III)-reducing conditions was investigated by compound specific stable isotope analysis (CSIA) using three model cultures (Aromatoleum aromaticum EbN1, Georgfuchsia toluolica G5G6, and a Azoarcus-dominated mixed culture). Systematically lower isotope enrichment factors for carbon and hydrogen were observed for particulate Mn(IV). The increasing diffusion distances of toluene or ethylbenzene to the solid Mn(IV) most likely caused limited bioavailability and hence resulted in the observed masking effect. The data suggests further ethylbenzene hydroxylation by ethylbenzene dehydrogenase (EBDH) and toluene activation by benzylsuccinate synthase (BSS) as initial activation steps. Notably, significantly different values in dual isotope analysis were detected for toluene degradation by G. toluolica under the three studied redox conditions, suggesting variations in the enzymatic transition state depending on the available TEA. The results indicate that two-dimensional CSIA has significant potential to assess anaerobic biodegradation of ethylbenzene and toluene at contaminated sites. Copyright © 2015 Elsevier Ltd. All rights reserved.

  11. 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.

  12. Isolation from soils ofNitrobacter and evidence for novel serotypes using immunofluorescence.

    Science.gov (United States)

    Josserand, A; Cleyet-Marel, J C

    1979-09-01

    To study the ecology of chemoautotrophic nitrifying bacteria (Nitrobacter), the immunofluorescence technique has been used. Fluorescent antibodies againstNitrobacter winogradskyi andNitrobacter agilis, the two known serotypes, have not labeled strains isolated from soils of the Lyon region (pH 8.1 and pH 4.7). The pure-culture isolates appeared to belong to the same genus, but to be serologically different from the reference strains. These results led us to question the diversity of strains ofNitrobacter in soils.

  13. 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

    BACKGROUND: Antinuclear antibodies (ANAs) are associated with several inflammatory rheumatic diseases. The aim of the present work was to evaluate enzyme immunoassays (EIAs) and compare them with classic immunofluorescent analysis (IFA) for the detection of ANA. METHODS: Seven enzyme immunoassays...... an IFA was performed (n = 164); and (c) a population of consecutive outpatients suspected to have a rheumatic disease (n = 101). The current clinical diagnoses of the patients served as the standard against which performance of the assays was evaluated. RESULTS: In patients with well...... and was mostly clinical setting in which...

  14. Localization of stationary pronuclei during conjugation of Paramecium as indicated by immunofluorescence staining.

    Science.gov (United States)

    Gao, Xin; Zhu, Jia-Jun; Yang, Xian-Yu; Yuan, Jin-Qiang; Wang, Yi-Wen; Song, Min-Guo

    2011-08-01

    After the third prezygotic division during conjugation of Paramecium caudatum, migratory and stationary pronuclei are produced. The migratory pronuclei remain in the paroral region tightly against the conjugating boundaries; while the stationary pronuclei are located beside the migratory pronuclei. To date, however, it is not clear what causes this close side-by-side localization between migratory and stationary pronuclei. In the current study, immunofluorescence staining with monoclonal antibody of anti-α tubulin indicated that ''U'' or ''V'' shaped spindles connected the migratory and stationary pronuclei during the third prezygotic division. This observation accounts for the close localization between these two types of pronuclei.

  15. Optimized Protocol of Methanol Treatment for Immunofluorescent Staining in Fixed Brain Slices.

    Science.gov (United States)

    Yuan, Feng; Xiong, Guoxiang; Cohen, Noam A; Cohen, Akiva S

    2017-03-01

    We optimized methanol treatment in paraformaldehyde-fixed slices for immunofluorescent staining of ependymal basal bodies in brain ventricles. As 100% methanol induced severe deformations to the slices (including rolling and folding over), we tried to decrease methanol concentration. We found that 33.3% to 75% methanol could result in ideal immunostaining of basal bodies without inducing obvious deformations. Instead of treating slices at -20°C (without proper cryoprotection measurements) as suggested in previous studies, we carried out methanol treatment at room temperature. Our modified protocol can not only raise immunostaining efficiency in tissue slices, it may also prevent potential freezing damages to the samples.

  16. Identification of cancer specific ligands from one-bead one compound combinatorial libraries to develop theranostics agents against oral squamous cell carcinoma

    Science.gov (United States)

    Yang, Frances Fan

    Background: Oral squamous cell carcinoma (OSCC) is one of the most prevalent disease worldwide. One-bead one-compound (OBOC) combinatorial technology is a powerful method to identify peptidomimetic ligands against a variety of receptors on cell surfaces. We therefore hypothesized that cancer specific ligands against OSCC might be identified and can be conjugated to optical dyes or nanocarriers to develop theranostic agents against OSCC. Material and methods: Different OSCC cell lines were incubated with OBOC libraries and beads with cell binding were sorted and then screened with normal human cells to identify peptide-beads binding to different OSCC cell lines but not binding to normal human cells. The molecular probes of OSCC were developed by biotinylating the carboxyl end of the ligands. OSCC theranostic agents were developed by decorating LLY13 with NPs and evaluated by using orthotopic bioluminescent oral cancer model. Results: Six OSCC specific ligands were discovered. Initial peptide-histochemistry study indicated that LLY12 and LLY13 were able to specifically detect OSCC cells grown on chamber slides at the concentration of 1 muM. In addition, LLY13 was found to penetrate into the OSCC cells and accumulate in the cytoplasm, and nucleus. After screened with a panel of integrin antibodies, only anti-alpha3 antibody was able to block most of OSCC cells binding to the LLY13 beads. OSCC theranostic agents developed using targeting LLY13 micelles (25+/- 4nm in diameter) were more efficient in binding to HSC-3 cancer cells compared to non-targeting micelles. Ex vivo images demonstrated that xenografts from the mice with targeting micelles appeared to have higher signals than the non-targeting groups. Conclusion: LLY13 has promising in vitro and in vivo targeting activity against OSCC. In addition, LLY13 is also able to penetrate into cancer cells via endocytosis. Initial study indicated that alpha3 integrin might partially be the corresponding receptor involved

  17. 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

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

    Science.gov (United States)

    Kishen, Ria E. B.; Kluth, David C.; Bellamy, Christopher O. C.

    2016-01-01

    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. PMID:27632367

  19. Histologic, ultrastructural, and immunofluorescent evaluation of human laser-assisted in situ keratomileusis corneal wounds.

    Science.gov (United States)

    Dawson, Daniel G; Kramer, Theresa R; Grossniklaus, Hans E; Waring, George O; Edelhauser, Henry F

    2005-06-01

    To evaluate human corneas after laser-assisted in situ keratomileusis at different postoperative intervals. Thirty-eight postmortem corneas from 20 patients with postoperative intervals from 2 months to 6.5 years after laser-assisted in situ keratomileusis surgery were collected from eye banks. The corneas were trisected and processed for conventional histologic analysis, transmission electron microscopy, and immunofluorescence. Light microscopy and transmission electron microscopy showed focal undulations in Bowman layer, focal epithelial hypertrophic modifications, and a variably thick (range, 0.4-16.4-mum) lamellar stromal interface scar in all specimens. The flap wound margin, which was adjacent to the epithelium, healed by producing an approximately 8-mum-thick hypercellular fibrotic stromal scar, whereas the central and paracentral wound regions healed differently because a thinner (approximately 5-mum) hypocellular primitive stromal scar was present in all the corneas examined. Immunofluorescence identified increased type 3 collagen and myofibroblasts in the hypercellular fibrotic scar regions and decreased or absent levels of all corneal stromal components other than type 1 collagen in the hypocellular primitive scar regions. After laser-assisted in situ keratomileusis surgery, the keratocyte-mediated production of a variably thick lamellar corneal stromal scar occurs, resulting in 2 regional types of scarring. The hypercellular fibrotic scar at the wound margin is usually visible clinically and functions to hold the flap in place, while the more central hypocellular primitive scar is not visible clinically and allows easy lifting of the flap postoperatively.

  20. The use of Biochip immunofluorescence microscopy for the serological diagnosis of epidermolysis bullosa acquisita.

    Science.gov (United States)

    Marzano, Angelo Valerio; Cozzani, Emanuele; Biasin, Matteo; Russo, Irene; Alaibac, Mauro

    2016-05-01

    Epidermolysis bullosa acquisita is a rare autoimmune bullous disease characterized by the presence of circulating antibodies directed against the collagen type VII. Diagnosis is generally based on clinical history, clinical features, histology, direct and indirect immunofluorescence, immunoblotting and ELISA. Our study aims to determine the validity of the Biochip immunofluorescence microscopy for the serological diagnosis of epidermolysis bullosa acquisita. Six patients with epidermolysis bullosa acquisita and presence of antibodies against type VII collagen confirmed by ELISA were included in the study. Subsequently, all sera of patients were analyzed using Biochip. Antibodies anti-collagen type VII were detected in all sera by means of the Biochip technology. Thus, Biochip shows a good correlation with ELISA and seems to be an appropriate method for the diagnosis of epidermolysis bullosa acquisita. It is an easy, fast and standardized method which could facilitate the diagnosis of this autoimmune bullous disease. We suggest that it could be used as an initial screening test to identify patients with epidermolysis bullosa acquisita.

  1. Direct and indirect immunofluorescence staining of fecal streptococci for rapid assessment of water quality

    Energy Technology Data Exchange (ETDEWEB)

    Pavlova, M.T.; Beauvais, E.; Brezenski, F.T.; Litsky, W.

    1975-01-01

    Immunofluorescence (IF) techniques were employed in an attempt to develop a rapid test for the identification of fecal streptococci. Fresh isolates were obtained from river waters and raw sewage. Identification to species were made by the conventional physiological, biochemical, and serological tests. Both whole and disrupted cells of representative strains of each species were used for the preparation of the fecal streptococcal vaccine. Globulin fractions of individual and pooled antisera were labeled with fluorescein isothiocyanate, and the resulting conjugates were tested with homologous and heterologous antigens. The present findings suggest that the immunofluorescence techniques can be employed in the determination of the presence and source of fecal pollution in water employing the fecal streptococci as indicator organisms. By using this method it was determined that fecal streptococci can be identified from water and sewage samples within 20 hours. Parenthetically it should be noted that the identification procedures using the routine biochemical and serological tests may take as long as 7 to 14 days. The procedure may be automated for continual monitoring.

  2. Clinical Implications of Direct Immunofluorescence Findings in Patients With Ocular Mucous Membrane Pemphigoid.

    Science.gov (United States)

    Labowsky, Mary T; Stinnett, Sandra S; Liss, Jason; Daluvoy, Melissa; Hall, Russell P; Shieh, Christine

    2017-11-01

    To examine the clinical implications of positive or negative direct immunofluorescence biopsies (DIF) in patients with clinically typical ocular mucous membrane pemphigoid (MMP). Retrospective cohort study. The study population was patients with clinically typical ocular MMP disease with documented DIF results who were followed for at least 1 year at the Duke University multidisciplinary ocular MMP clinic. Data were collected by chart review and included patient demographics, clinical examination findings, and history of autoimmune disease and/or malignancy, as well as topical, systemic, and surgical treatments received. Main outcome measures included MMP Disease Area Index, Foster stages, proportion legally blind, duration of follow-up, and use of systemic immunosuppression and ocular procedures in treatment. In multivariable analysis restricted to 55 patients, patients with negative and positive biopsies were similar in the outcome measures; however, positive-biopsy patients were more likely to be treated with systemic immunosuppression and were followed for longer at our clinic. Patients with isolated ocular disease were also more likely to have negative biopsies compared to those who also had extraocular disease. Patients who had conjunctival biopsies were more likely to have a negative direct immunofluorescence result than patients with biopsies from other sites. We encourage clinicians and patients to consider treatment with systemic immunosuppression even in the absence of diagnosis confirmation by DIF. Furthermore, this study supports current standard of care to pursue a nonocular biopsy of normal-appearing, perilesional skin or oral mucosa when possible. Published by Elsevier Inc.

  3. Delay, change and bifurcation of the immunofluorescence distribution attractors in health statuses diagnostics and in medical treatment

    Science.gov (United States)

    Galich, Nikolay E.; Filatov, Michael V.

    2008-07-01

    Communication contains the description of the immunology experiments and the experimental data treatment. New nonlinear methods of immunofluorescence statistical analysis of peripheral blood neutrophils have been developed. We used technology of respiratory burst reaction of DNA fluorescence in the neutrophils cells nuclei due to oxidative activity. The histograms of photon count statistics the radiant neutrophils populations' in flow cytometry experiments are considered. Distributions of the fluorescence flashes frequency as functions of the fluorescence intensity are analyzed. Statistic peculiarities of histograms set for healthy and unhealthy donors allow dividing all histograms on the three classes. The classification is based on three different types of smoothing and long-range scale averaged immunofluorescence distributions and their bifurcation. Heterogeneity peculiarities of long-range scale immunofluorescence distributions allow dividing all histograms on three groups. First histograms group belongs to healthy donors. Two other groups belong to donors with autoimmune and inflammatory diseases. Some of the illnesses are not diagnosed by standards biochemical methods. Medical standards and statistical data of the immunofluorescence histograms for identifications of health and illnesses are interconnected. Possibilities and alterations of immunofluorescence statistics in registration, diagnostics and monitoring of different diseases in various medical treatments have been demonstrated. Health or illness criteria are connected with statistics features of immunofluorescence histograms. Neutrophils populations' fluorescence presents the sensitive clear indicator of health status.

  4. Nutrient regime shift in the western North Atlantic indicated by compound-specific δ15N of deep-sea gorgonian corals.

    Science.gov (United States)

    Sherwood, Owen A; Lehmann, Moritz F; Schubert, Carsten J; Scott, David B; McCarthy, Matthew D

    2011-01-18

    Despite the importance of the nitrogen (N) cycle on marine productivity, little is known about variability in N sources and cycling in the ocean in relation to natural and anthropogenic climate change. Beyond the last few decades of scientific observation, knowledge depends largely on proxy records derived from nitrogen stable isotopes (δ(15)N) preserved in sediments and other bioarchives. Traditional bulk δ(15)N measurements, however, represent the combined influence of N source and subsequent trophic transfers, often confounding environmental interpretation. Recently, compound-specific analysis of individual amino acids (δ(15)N-AA) has been shown as a means to deconvolve trophic level versus N source effects on the δ(15)N variability of bulk organic matter. Here, we demonstrate the first use of δ(15)N-AA in a paleoceanographic study, through analysis of annually secreted growth rings preserved in the organic endoskeletons of deep-sea gorgonian corals. In the Northwest Atlantic off Nova Scotia, coral δ(15)N is correlated with increasing presence of subtropical versus subpolar slope waters over the twentieth century. By using the new δ(15)N-AA approach to control for variable trophic processing, we are able to interpret coral bulk δ(15)N values as a proxy for nitrate source and, hence, slope water source partitioning. We conclude that the persistence of the warm, nutrient-rich regime since the early 1970s is largely unique in the context of the last approximately 1,800 yr. This evidence suggests that nutrient variability in this region is coordinated with recent changes in global climate and underscores the broad potential of δ(15)N-AA for paleoceanographic studies of the marine N cycle.

  5. Involvement of the Leaf-Specific Multidrug and Toxic Compound Extrusion (MATE) Transporter Nt-JAT2 in Vacuolar Sequestration of Nicotine in Nicotiana tabacum

    Science.gov (United States)

    Shitan, Nobukazu; Minami, Shota; Morita, Masahiko; Hayashida, Minaho; Ito, Shingo; Takanashi, Kojiro; Omote, Hiroshi; Moriyama, Yoshinori; Sugiyama, Akifumi; Goossens, Alain; Moriyasu, Masataka; Yazaki, Kazufumi

    2014-01-01

    Alkaloids play a key role in higher plant defense against pathogens and herbivores. Following its biosynthesis in root tissues, nicotine, the major alkaloid of Nicotiana species, is translocated via xylem transport toward the accumulation sites, leaf vacuoles. Our transcriptome analysis of methyl jasmonate-treated tobacco BY-2 cells identified several multidrug and toxic compound extrusion (MATE) transporter genes. In this study, we characterized a MATE gene, Nicotiana tabacum jasmonate-inducible alkaloid transporter 2 (Nt-JAT2), which encodes a protein that has 32% amino acid identity with Nt-JAT1. Nt-JAT2 mRNA is expressed at a very low steady state level in whole plants, but is rapidly upregulated by methyl jasmonate treatment in a leaf-specific manner. To characterize the function of Nt-JAT2, yeast cells were used as the host organism in a cellular transport assay. Nt-JAT2 was localized at the plasma membrane in yeast cells. When incubated in nicotine-containing medium, the nicotine content in Nt-JAT2-expressing cells was significantly lower than in control yeast. Nt-JAT2-expressing cells also showed lower content of other alkaloids like anabasine and anatabine, but not of flavonoids, suggesting that Nt-JAT2 transports various alkaloids including nicotine. Fluorescence assays in BY-2 cells showed that Nt-JAT2-GFP was localized to the tonoplast. These findings indicate that Nt-JAT2 is involved in nicotine sequestration in leaf vacuoles following the translocation of nicotine from root tissues. PMID:25268729

  6. Direct photolysis of sulfamethoxazole using various irradiation sources and wavelength ranges - insights from degradation product analysis and compound-specific stable isotope analysis.

    Science.gov (United States)

    Willach, Sarah; Lutze, Holger Volker; Eckey, Kevin; Löppenberg, Katja; Lueling, Michelle; Wolbert, Jens-Benjamin; Kujawinski, Dorothea M; Jochmann, Maik A; Karst, Uwe; Schmidt, Torsten Claus

    2018-01-05

    The environmental micropollutant sulfamethoxazole (SMX) is susceptible to phototransformation by sunlight and UV-C light which is used for water disinfection. Depending on the environmental pH conditions SMX may be present as neutral or anionic species. This study systematically investigates the phototransformation of these two relevant SMX species using four different irradiation scenarios, i.e. a low, medium and high pressure Hg lamp and simulated sunlight. The observed phototransformation kinetics are complemented by data from compound-specific stable isotope and transformation product analysis using isotope-ratio and high-resolution mass spectrometry (HRMS). Observed phototransformation kinetics were faster for the neutral than for the anionic SMX species (from 3.4 (LP lamp) up to 6.6 (HP lamp) times). Furthermore, four phototransformation products (with m/z 189, 202, 242, and 260) were detected by HRMS that have not yet been described for direct photolysis of SMX. Isotopic fractionation occurred only if UV-B and UV-A wavelengths prevailed in the emitted irradiation and was most pronounced for the neutral species with simulated sunlight (εC = 4.8 ± 0.1 ‰). Phototransformation of SMX with UV-C light did not cause significant isotopic fractionation. Consequently, it was possible to differentiate sunlight and UV-C light induced phototransformation of SMX. Thus CSIA might be implemented to trace back waste water point sources or to assess natural attenuation of SMX by sunlight photolysis. In contrast to the wavelength range, pH-dependent speciation of SMX did hardly impact isotopic fractionation.

  7. All fats are not equal: Considerations when using fatty acid biomarkers in compound-specific stable isotope soil and sediment tracing

    Science.gov (United States)

    Reiffarth, Dominic; Petticrew, Ellen; Owens, Philip; Lobb, David

    2013-04-01

    The development of cost-effective, convenient and reliable methods for tracing sediment movement will help establish water security. The use of compound-specific stable isotopes (CSSIs) has seen limited, small-scale applications, often in watersheds exhibiting exotic and highly diverse vegetation types. The CSSI tracing technique relies on the detection and transport of naturally occurring organics of plant origin (biomarkers); the biomarkers of interest are produced by flora, deposited on the soil and potentially mobilized along with soil particles. In part, the uniqueness of a biomarker is dependent on its biological pathway. As a plant fixes CO2-its primary source of carbon for building larger organic molecules-discrimination against the heavier 13C isotope leads to an enrichment of 12C. The more complex the biological pathway the biomarker has been subjected to, the more discrimination and unique isotopic signature the biomarker exhibits. Successfully implementing CSSI tracing requires recognizing: (i) factors contributing to the natural variability of the isotopic signature (ii) the suitability of a biomarker and (iii) factors affecting sensitivity during analysis. Considering the relatively low input of suitable organic biomarkers into the soil and degree of signal dispersion, care must be taken to isolate and correctly identify biomarkers, particularly where vegetation types show low variability and where long-range transport occurs. Research is currently being conducted in the Horsefly River (British Columbia, Canada) and South Tobacco Creek (Manitoba, Canada) watersheds; the research seeks to address some of these concerns, particularly in a temperate climate where exotic vegetation types are not common and variability is expected to be low.

  8. Implications for chloro- and pheopigment synthesis and preservation from combined compound-specific δ13C, δ15N, and Δ14C analysis

    Directory of Open Access Journals (Sweden)

    G. Mollenhauer

    2010-12-01

    Full Text Available Chloropigments and their derivative pheopigments preserved in sediments can directly be linked to photosynthesis. Their carbon and nitrogen stable isotopic compositions have been shown to be a good recorder of recent and past surface ocean environmental conditions tracing the carbon and nitrogen sources and dominant assimilation processes of the phytoplanktonic community. In this study we report results from combined compound-specific radiocarbon and stable carbon and nitrogen isotope analysis to examine the time-scales of synthesis and fate of chlorophyll-a and its degradation products pheophytin-a, pyropheophytin-a, and 132,173-cyclopheophorbide-a-enol until burial in Black Sea core-top sediments. The pigments are mainly of marine phytoplanktonic origin as implied by their stable isotopic compositions. Pigment δ15N values indicate nitrate as the major uptake substrate but 15N-depletion towards the open marine setting indicates either contribution from N2-fixation or direct uptake of ammonium from deeper waters. Radiocarbon concentrations translate into minimum and maximum pigment ages of approximately 40 to 1200 years. This implies that protective mechanisms against decomposition such as association with minerals, storage in deltaic anoxic environments, or eutrophication-induced hypoxia and light limitation are much more efficient than previously thought. Moreover, seasonal variations of nutrient source, growth period, and habitat and their associated isotopic variability are likely at least as strong as long-term trends. Combined triple isotope analysis of sedimentary chlorophyll and its primary derivatives is a powerful tool to delineate biogeochemical and diagenetic processes in the surface water and sediments, and to assess their precise time-scales.

  9. Evaluation of ethyl tert-butyl ether biodegradation in a contaminated aquifer by compound-specific isotope analysis and in situ microcosms

    Energy Technology Data Exchange (ETDEWEB)

    Bombach, Petra, E-mail: petra.bombach@ufz.de [UFZ – Helmholtz Centre for Environmental Research, Department of Isotope Biogeochemistry, Permoserstrasse 15, D-04318 Leipzig (Germany); Isodetect GmbH Leipzig, Deutscher Platz 5b, D-04103 Leipzig (Germany); Nägele, Norbert [Kuvier the Biotech Company S.L., Ctra. N-I, p.k. 234–P.E. INBISA 23" a, E-09001 Burgos (Spain); Rosell, Mònica [UFZ – Helmholtz Centre for Environmental Research, Department of Isotope Biogeochemistry, Permoserstrasse 15, D-04318 Leipzig (Germany); Grup de Mineralogia Aplicada i Medi Ambient, Departament de Cristallografia, Mineralogia i Dipòsits Minerals, Facultat de Geologia, Universitat de Barcelona (UB), C/Martí i Franquès s/n, 08028 Barcelona (Spain); Richnow, Hans H. [UFZ – Helmholtz Centre for Environmental Research, Department of Isotope Biogeochemistry, Permoserstrasse 15, D-04318 Leipzig (Germany); Fischer, Anko [Isodetect GmbH Leipzig, Deutscher Platz 5b, D-04103 Leipzig (Germany)

    2015-04-09

    Highlights: • In situ biodegradation of ETBE was investigated in a fuel contaminated aquifer. • Degradation was studied by CSIA and in situ microcosms in combination with TLFA-SIP. • ETBE was degraded when ETBE was the main groundwater contaminant. • ETBE was also degraded in the presence of BTEX and MTBE. • Hydrochemical analysis indicated aerobic and anaerobic ETBE biodegradation. - Abstract: Ethyl tert-butyl ether (ETBE) is an upcoming groundwater pollutant in Europe whose environmental fate has been less investigated, thus far. In the present study, we investigated the in situ biodegradation of ETBE in a fuel-contaminated aquifer using compound-specific stable isotope analysis (CSIA), and in situ microcosms in combination with total lipid fatty acid (TLFA)-stable isotope probing (SIP). In a first field investigation, CSIA revealed insignificant carbon isotope fractionation, but low hydrogen isotope fractionation of up to +14‰ along the prevailing anoxic ETBE plume suggesting biodegradation of ETBE. Ten months later, oxygen injection was conducted to enhance the biodegradation of petroleum hydrocarbons (PH) at the field site. Within the framework of this remediation measure, in situ microcosms loaded with [{sup 13}C{sub 6}]-ETBE (BACTRAP{sup ®}s) were exposed for 119 days in selected groundwater wells to assess the biodegradation of ETBE by TLFA-SIP under the following conditions: (i) ETBE as main contaminant; (ii) ETBE as main contaminant subjected to oxygen injection; (iii) ETBE plus other PH; (iv) ETBE plus other PH subjected to oxygen injection. Under all conditions investigated, significant {sup 13}C-incorporation into microbial total lipid fatty acids extracted from the in situ microcosms was found, providing clear evidence of ETBE biodegradation.

  10. The application of compound-specific isotope analysis of fatty acids for traceability of sea cucumber (Apostichopus japonicus) in the coastal areas of China.

    Science.gov (United States)

    Liu, Yu; Zhang, Xufeng; Li, Ying; Wang, Haixia

    2017-11-01

    Geographical origin traceability is an important issue for controlling the quality of seafood and safeguarding the interest of consumers. In the present study, a new method of compound-specific isotope analysis (CSIA) of fatty acids was established to evaluate its applicability in establishing the origin traceability of Apostichopus japonicus in the coastal areas of China. Moreover, principal component analysis (PCA) and discriminant analysis (DA) were applied to distinguish between the origins of A. japonicus. The results show that the stable carbon isotope compositions of fatty acids of A. japonicus significantly differ in terms of both season and origin. They also indicate that the stable carbon isotope composition of fatty acids could effectively discriminate between the origins of A. japonicus, except for between Changhai Island and Zhangzi Island in the spring of 2016 because of geographical proximity or the similarity of food sources. The fatty acids that have the highest contribution to identifying the geographical origins of A. japonicus are C22:6n-3, C16:1n-7, C20:5n-3, C18:0 and C23:1n-9, when considering the fatty acid contents, the stable carbon isotope composition of fatty acids and the results of the PCA and DA. We conclude that CSIA of fatty acids, combined with multivariate statistical analysis such as PCA and DA, may be an effective tool for establishing the traceability of A. japonicus in the coastal areas of China. The relevant conclusions of the present study provide a new method for determining the traceability of seafood or other food products. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  11. Metabolism of Seriola lalandi during Starvation as Revealed by Fatty Acid Analysis and Compound-Specific Analysis of Stable Isotopes within Amino Acids.

    Directory of Open Access Journals (Sweden)

    Fernando Barreto-Curiel

    Full Text Available Fish starvation is defined as food deprivation for a long period of time, such that physiological processes become confined to basal metabolism. Starvation provides insights in physiological processes without interference from unknown factors in digestion and nutrient absorption occurring in fed state. Juveniles of amberjack Seriola lalandi were isotopically equilibrated to a formulated diet for 60 days. One treatment consisted of fish that continued to be fed and fish in the other treatment were not fed for 35 days. The isotopic signatures prior to the beginning of and after the starvation period, for fish in the starvation and control treatments, were analysed for lipid content, fatty acid composition and isotopic analysis of bulk (EA-IRMS and of amino acids (compound specific isotope analysis, CSIA. There were three replicates for the starvation group. Fatty acid content in muscle and liver tissue before and after starvation was determined to calculate percent change. Results showed that crude lipid was the most used source of energy in most cases; the PUFAs and LC-PUFAs were highly conserved. According to the protein signature in bulk (δ15N and per amino acid (δ13C and δ15N, in muscle tissue, protein synthesis did not appear to occur substantially during starvation, whereas in liver, increases in δ13C and δ15N indicate that protein turnover occurred, probably for metabolic routing to energy-yielding processes. As a result, isotopic values of δ15N in muscle tissue do not change, whereas CSIA net change occurred in the liver tissue. During the study period of 35 days, muscle protein was largely conserved, being neither replenished from amino acid pools in the plasma and liver nor catabolized.

  12. Characterization of sulfur compounds in whisky by full evaporation dynamic headspace and selectable one-dimensional/two-dimensional retention time locked gas chromatography-mass spectrometry with simultaneous element-specific detection.

    Science.gov (United States)

    Ochiai, Nobuo; Sasamoto, Kikuo; MacNamara, Kevin

    2012-12-28

    A method is described for characterization of sulfur compounds in unaged and aged whisky. The method is based on full evaporation dynamic headspace (FEDHS) of 100 μL of whisky samples followed by selectable one-dimensional ((1)D) or two-dimensional ((2)D) retention-time-locked (RTL) gas chromatography (GC)-mass spectrometry (MS) with simultaneous element-specific detection using a sulfur chemiluminescence detector (SCD) and a nitrogen chemiluminescence detector (NCD). Sequential heart-cuts of the 16 sulfur fractions were used to identify each individual sulfur compound in the unaged whisky. Twenty sulfur compounds were positively identified by a MS library search, linear retention indices (LRI), and formula identification using MS calibration software. Additionally eight formulas were also identified for unknown sulfur compounds. Simultaneous heart-cuts of the 16 sulfur fractions were used to produce the (2)D RTL GC-SCD chromatograms for principal component analysis. PCA of the (2)D RTL GC-SCD data clearly demonstrated the difference between unaged and aged whisky, as well as two different whisky samples. Fourteen sulfur compounds could be characterized as key sulfur compounds responsible for the changes in the aging step and/or the difference between two kinds of whisky samples. The determined values of the key sulfur compounds were in the range of 0.3-210 ng mL(-1) (RSD: 0.37-12%, n=3). Copyright © 2012 Elsevier B.V. All rights reserved.

  13. Sensitive and Quantitative Detection of C-Reaction Protein Based on Immunofluorescent Nanospheres Coupled with Lateral Flow Test Strip.

    Science.gov (United States)

    Hu, Jiao; Zhang, Zhi-Ling; Wen, Cong-Ying; Tang, Man; Wu, Ling-Ling; Liu, Cui; Zhu, Lian; Pang, Dai-Wen

    2016-06-21

    Sensitive and quantitative detection of protein biomarkers with a point-of-care (POC) assay is significant for early diagnosis, treatment, and prognosis of diseases. In this paper, a quantitative lateral flow assay with high sensitivity for protein biomarkers was established by utilizing fluorescent nanospheres (FNs) as reporters. Each fluorescent nanosphere (FN) contains 332 ± 8 CdSe/ZnS quantum dots (QDs), leading to its superstrong luminescence, 380-fold higher than that of one QD. Then a detection limit of 27.8 pM C-reaction protein (CRP) could be achieved with an immunofluorescent nanosphere (IFN)-based lateral flow test strip. The assay was 257-fold more sensitive than that with a conventional Au-based lateral flow test strip for CRP detection. Besides, the fluorescence intensity of FNs and bioactivity of IFNs were stable during 6 months of storage. Hence, the assay owns good reproducibility (intra-assay variability of 5.3% and interassay variability of 6.6%). Furthermore, other cancer biomarkers (PSA, CEA, AFP) showed negative results by this method, validating the excellent specificity of the method. Then the assay was successfully applied to quantitatively detect CRP in peripheral blood plasma samples from lung cancer and breast cancer patients, and healthy people, facilitating the diagnosis of lung cancer. It holds a good prospect of POC protein biomarker detection.

  14. Reversible optical control of cyanine fluorescence in fixed and living cells: optical lock-in detection immunofluorescence imaging microscopy.

    Science.gov (United States)

    Yan, Yuling; Petchprayoon, Chutima; Mao, Shu; Marriott, Gerard

    2013-02-05

    Optical switch probes undergo rapid and reversible transitions between two distinct states, one of which may fluoresce. This class of probe is used in various super-resolution imaging techniques and in the high-contrast imaging technique of optical lock-in detection (OLID) microscopy. Here, we introduce optimized optical switches for studies in living cells under standard conditions of cell culture. In particular, a highly fluorescent cyanine probe (Cy or Cy3) is directly or indirectly linked to naphthoxazine (NISO), a highly efficient optical switch that undergoes robust, 405/532 nm-driven transitions between a colourless spiro (SP) state and a colourful merocyanine (MC) state. The intensity of Cy fluorescence in these Cy/Cy3-NISO probes is reversibly modulated between a low and high value in SP and MC states, respectively, as a result of Förster resonance energy transfer. Cy/Cy3-NISO probes are targeted to specific proteins in living cells where defined waveforms of Cy3 fluorescence are generated by optical switching of the SP and MC states. Finally, we introduce a new imaging technique (called OLID-immunofluorescence microscopy) that combines optical modulation of Cy3 fluorescence from Cy3/NISO co-labelled antibodies within fixed cells and OLID analysis to significantly improve image contrast in samples having high background or rare antigens.

  15. 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.

  16. Integration of co-localized glandular morphometry and protein biomarker expression in immunofluorescent images for prostate cancer prognosis

    Science.gov (United States)

    Scott, Richard; Khan, Faisal M.; Zeineh, Jack; Donovan, Michael; Fernandez, Gerardo

    2015-03-01

    Immunofluorescent (IF) image analysis of tissue pathology has proven to be extremely valuable and robust in developing prognostic assessments of disease, particularly in prostate cancer. There have been significant advances in the literature in quantitative biomarker expression as well as characterization of glandular architectures in discrete gland rings. However, while biomarker and glandular morphometric features have been combined as separate predictors in multivariate models, there is a lack of integrative features for biomarkers co-localized within specific morphological sub-types; for example the evaluation of androgen receptor (AR) expression within Gleason 3 glands only. In this work we propose a novel framework employing multiple techniques to generate integrated metrics of morphology and biomarker expression. We demonstrate the utility of the approaches in predicting clinical disease progression in images from 326 prostate biopsies and 373 prostatectomies. Our proposed integrative approaches yield significant improvements over existing IF image feature metrics. This work presents some of the first algorithms for generating innovative characteristics in tissue diagnostics that integrate co-localized morphometry and protein biomarker expression.

  17. Inferring Phytoplankton, Terrestrial Plant and Bacteria Bulk δ¹³C Values from Compound Specific Analyses of Lipids and Fatty Acids

    Science.gov (United States)

    Taipale, Sami J.; Peltomaa, Elina; Hiltunen, Minna; Jones, Roger I.; Hahn, Martin W.; Biasi, Christina; Brett, Michael T.

    2015-01-01

    Stable isotope mixing models in aquatic ecology require δ13C values for food web end members such as phytoplankton and bacteria, however it is rarely possible to measure these directly. Hence there is a critical need for improved methods for estimating the δ13C ratios of phytoplankton, bacteria and terrestrial detritus from within mixed seston. We determined the δ13C values of lipids, phospholipids and biomarker fatty acids and used these to calculate isotopic differences compared to the whole-cell δ13C values for eight phytoplankton classes, five bacterial taxa, and three types of terrestrial organic matter (two trees and one grass). The lipid content was higher amongst the phytoplankton (9.5±4.0%) than bacteria (7.3±0.8%) or terrestrial matter (3.9±1.7%). Our measurements revealed that the δ13C values of lipids followed phylogenetic classification among phytoplankton (78.2% of variance was explained by class), bacteria and terrestrial matter, and there was a strong correlation between the δ13C values of total lipids, phospholipids and individual fatty acids. Amongst the phytoplankton, the isotopic difference between biomarker fatty acids and bulk biomass averaged -10.7±1.1‰ for Chlorophyceae and Cyanophyceae, and -6.1±1.7‰ for Cryptophyceae, Chrysophyceae and Diatomophyceae. For heterotrophic bacteria and for type I and type II methane-oxidizing bacteria our results showed a -1.3±1.3‰, -8.0±4.4‰, and -3.4±1.4‰ δ13C difference, respectively, between biomarker fatty acids and bulk biomass. For terrestrial matter the isotopic difference averaged -6.6±1.2‰. Based on these results, the δ13C values of total lipids and biomarker fatty acids can be used to determine the δ13C values of bulk phytoplankton, bacteria or terrestrial matter with ± 1.4‰ uncertainty (i.e., the pooled SD of the isotopic difference for all samples). We conclude that when compound-specific stable isotope analyses become more widely available, the determination of

  18. Specific features of chemical composition of neutral oxygen-, nitrogen-, and sulfur-containing compounds from tars of lignite semicoking in steps

    Energy Technology Data Exchange (ETDEWEB)

    Platonov, V.V.; Proskuryakov, V.A.; Pipchenko, I.A.; Novikova, I.L. [Tula State Pedagogical University, Tula (Russian Federation)

    1998-01-01

    Component and structural-group composition of neutral oxygen- nitrogen-, and sulfur-containing compounds isolated from tars of semicoking in steps of lignite from the Berezovo deposit of Kansk-Achinsk coal fields was studied.

  19. Immunofluorescent visualization of mouse interneuron subtypes [v2; ref status: indexed, http://f1000r.es/4qq

    Directory of Open Access Journals (Sweden)

    Simon Molgaard

    2014-11-01

    Full Text Available The activity of excitatory neurons is controlled by a highly diverse population of inhibitory interneurons. These cells show a high level of physiological, morphological and neurochemical heterogeneity, and play highly specific roles in neuronal circuits. In the mammalian hippocampus, these are divided into 21 different subtypes of GABAergic interneurons based on their expression of different markers, morphology and their electrophysiological properties. Ideally, all can be marked using an antibody directed against the inhibitory neurotransmitter GABA, but parvalbumin, calbindin, somatostatin, and calretinin are also commonly used as markers to narrow down the specific interneuron subtype. Here, we describe a journey to find the necessary immunological reagents for studying GABAergic interneurons of the mouse hippocampus. Based on web searches there are several hundreds of different antibodies on the market directed against these four markers. Searches in the literature databases allowed us to narrow it down to a subset of antibodies most commonly used in publications. However, in our hands the most cited ones did not work for immunofluorescence stainings of formaldehyde fixed tissue sections and cultured hippocampal neurons, and we had to immunostain our way through thirteen different commercial antibodies before finally finding a suitable antibody for each of the four markers. The antibodies were evaluated based on signal-to-noise ratios as well as if positive cells were found in layers of the hippocampus where they have previously been described. Additionally, the antibodies were also tested on sections from mouse spinal cord with similar criteria for specificity of the antibodies. Using the antibodies with a high rating on pAbmAbs, an antibody review database, stainings with high signal-to-noise ratios and location of the immunostained cells in accordance with the literature could be obtained, making these antibodies suitable choices for

  20. 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.

  1. Free radical scavenging capacity, anticandicidal effect of bioactive compounds from Sida cordifolia L., in combination with nystatin and clotrimazole and their effect on specific immune response in rats.

    Science.gov (United States)

    Ouédraogo, Maurice; Konaté, Kiessoun; Lepengué, Alexis Nicaise; Souza, Alain; M'Batchi, Bertrand; Sawadogo, Laya L

    2012-12-26

    Infectious diseases caused by fungi are still a major threat to public health, despite numerous efforts by researchers. Use of ethnopharmacological knowledge is one attractive way to reduce empiricism and enhance the probability of success in new drug-finding efforts. In this work, the total alkaloid compounds (AC) from Sida cordifolia L. (Malvaceae) have been investigated for their free radical scavenging capacity, antifungal and immunostimulatory properties. The antifungal activity was investigated against five candida strains using the microplate dilution method and the Fractional Inhibitory Concentration Index (FICI) of compounds was evaluated. The antioxidant activity of the samples was evaluate using three separate methods, at last, the immunostimulatory effect on immunosuppressed wistar rats was performed. As for the antifungal activity, result varied according to microorganism. The results obtained in this antifungal activity were interesting and indicated a synergistic effect between alkaloid compounds and the antifungal references such as Nystatin and Clotrimazole. Antioxidant capacity noticed that the reduction capacity of DPPH radicals obtained the best result comparatively to the others methods of free radical scavenging. Our results showed a low immunostimulatory effect and this result could be explained by the lack of biologically active antioxidants such as polyphenol compounds lowly contained in the alkaloid compounds. The results of this study showed that alkaloid compounds in combination with antifungal references (Nystatin and Clotrimazole) exhibited antimicrobial effects against candida strains tested. The results supported the utilization of these plants in infectious diseases particularly in treatment of candida infections.

  2. Pyrolysis compound specific isotopic analysis (δ13C and δD Py-CSIA) of soil organic matter size fractions under four vegetation covers.

    Science.gov (United States)

    Jiménez-Morillo, Nicasio T.; González-Vila, Francisco J.; Almendros, Gonzalo; De la Rosa, José M.; González-Pérez, José A.

    2015-04-01

    A chemical characterization of soil organic matter (SOM) under different ground cover from a Mediterranean climate (Doñana National Park, Andalusia, Spain) is approached using bulk δ15N, δ13C, δ18O and δD isotopic analysis (C/TC-IRMS) and δ13C and δD pyrolysis compound specific isotopic analysis (Py-CSIA: Py-GC-C/TC-IRMS). Soil samples were collected in sandy soils, Arenosols (WRB 2006) from the Doñana National Park (SW Spain) under different vegetation cover: cork oak (Quercus suber, QS), eagle fern (Pteridium aquilinum, PA), pine (Pinus pinea, PP) and rockrose (Halimium halimifolium, HH). Two size fractions; coarse (C: 1-2 mm) and fine (F: biological reworking with a higher microbiological activity fixing the lighter isotope in the soil fine organic fractions. It is known that lipid hydrogen is deuterium depleted relative to bulk organic hydrogen (Smith and Epstein, 1970). In line with this, in our study the lipid derived compounds had the largest deuterium depleted signature with a difference between bulk and lipid δD values was c. -35‰. This fractionation was highest in Pine (PP) and Rockrose (HH). The combination of traditional techniques for the study of SOM i.e. Py-GC/MS and IRMS, with new hyphenated analytical pyrolysis techniques i.e. Py-CSIA opens new possibilities and windows of information in SOM research. Our findings points to the occurrence of more or less complex processes that affects SOM chemical characteristics; whereas the coarse fraction resembles the chemical structure of the above vegetation, this SOM "memory" is less defined in the fine fractions, probably due to biologically mediated chemical conversions, additions and selective preservation of refractory chemical structures. SOM bulk IRMS and Py-CSIA are also informative about relevant plant physiological processes, OM sources and OM evolution status. Acknowledgments: N.T Jiménez-Morillo is funded by a FPI research grant (BES-2013-062573). Dr. J.M. de la Rosa is the recipient

  3. Triple immunofluorescence labeling of atherosclerotic plaque components in apoE/LDLR -/- mice.

    Science.gov (United States)

    Gajda, Mariusz; Jawień, Jacek; Mateuszuk, Lukasz; Lis, Grzegorz J; Radziszewski, Andrzej; Chłopicki, Stefan; Litwin, Jan A

    2008-01-01

    This paper presents a simple and reliable method of triple immunofluorescence staining that allows simultaneous detection of various cell types present in atherosclerotic plaque of apolipoprotein E and LDL receptor-double knockout (apoE/LDLR -/-) mice. We used combined direct and indirect procedures applying commercially available primary antibodies raised in different species to detect smooth muscle cells (Cy3-conjugated mouse anti-smooth muscle actin, SMA), macrophages (rat anti-CD68) and T lymphocytes (rabbit anti-CD3). Fixation of the material in acetone and modified incubation protocol employing nonfat dry milk in preincubation and incubation media significantly increased the intensity of labeling and effectively quenched the background. Our method offers an efficient way to detect qualitative as well as quantitative changes of macrophages, T lymphocytes and smooth muscle cells in atherosclerotic plaque of apoE/LDLR -/- mice during atherosclerosis development or in response to pharmacological treatment.

  4. Triple immunofluorescence labeling of atherosclerotic plaque components in apoE/LDLR -/- mice.

    Directory of Open Access Journals (Sweden)

    Stefan Chłopicki

    2008-06-01

    Full Text Available This paper presents a simple and reliable method of triple immunofluorescence staining that allows simultaneous detection of various cell types present in atherosclerotic plaque of apolipoprotein E and LDL receptor-double knockout (apoE/LDLR -/- mice. We used combined direct and indirect procedures applying commercially available primary antibodies raised in different species to detect smooth muscle cells (Cy3-conjugated mouse anti-smooth muscle actin, SMA, macrophages (rat anti-CD68 and T lymphocytes (rabbit anti-CD3. Fixation of the material in acetone and modified incubation protocol employing nonfat dry milk in preincubation and incubation media significantly increased the intensity of labeling and effectively quenched the background. Our method offers an efficient way to detect qualitative as well as quantitative changes of macrophages, T lymphocytes and smooth muscle cells in atherosclerotic plaque of apoE/LDLR -/- mice during atherosclerosis development or in response to pharmacological treatment.

  5. Direct immunofluorescence in granuloma faciale: a case report and review of literature.

    Science.gov (United States)

    Barnadas, M A; Curell, R; Alomar, A

    2006-07-01

    A 49-year-old male presented at our department with erythematous brownish plaques in the malar areas and left cheek of 9 years' evolution. Histopathological study revealed a dense, predominantly perivascular, inflammatory infiltrate in the reticular dermis. The infiltrate was composed of abundant neutrophils, lymphocytes, histiocytes, and eosinophils. Leukocytoclasia and fibrin in some vessel walls were also observed. The patient was diagnosed with granuloma faciale (GF). Direct immunofluorescence (DIF) study showed heavy immunoglobulin G (IgG) and less intense deposits of IgA, IgM, C(3) and C(1q) surrounding superficial and deep vessels of the reticular dermis. Perivascular and diffuse fibrinogen deposits were also present. No deposits were detected at the dermoepidermal junction. Our findings lend support to the hypothesis that classical pathway activation of complement may be involved in the development of vasculitis in GF.

  6. A rare case of signet ring cell lymphoma: Diagnosis aided by immunofluorescent staining

    Directory of Open Access Journals (Sweden)

    Charusheela R Gore

    2017-01-01

    Full Text Available Signet ring cell lymphomas are the proliferations of malignant lymphoid cells containing cytoplasmic vacuoles or globules which displace the nuclei, imparting it a signet ring appearance. This rare tumor is a variant of non-Hodgkin lymphoma. Signet ring appearance is due to cytoplasmic accumulation of immunoglobulin or vacuoles derived from multivesicular bodies. These cells, particularly with cytoplasmic vacuoles, may be mistaken for adenocarcinoma cells. We are presenting one such case where immunofluorescence helped us to demonstrate the immunoglobulins on fine needle aspiration smears. This is an innovative technique and has not been reported earlier. Our aim of presenting this case is to review the awareness of this rare lymphoma among pathologists to give due consideration for avoiding inappropriate investigations and treatment.

  7. Studies on bluetongue. V. Detection of the virus in infected materials by immunofluorescence.

    Science.gov (United States)

    Ruckerbauer, G M; Gray, D P; Girard, A; Bannister, G L; Boulanger, P

    1967-07-01

    The fluorescent-antibody technique was employed for the detection of bluetongue virus in bovine foetal kidney cell cultures inoculated with tissues and blood from experimentally infected animals. In the first series, a total number of 79 inoculated suckling-mouse brains were examined, 36 as frozen sections alone and 43 as impression slides in conjunction with tissue culture inoculation of the same material. With the combined tissue culture immunofluorescent methods, 36 suspicious were detected by impression smears and 37 positives by the tissue culture out of 43 brains examined. Twenty-two were suspicious out of the 36 examined as frozen sections. Results obtained with the second series, using sheep tissues, showed that the combined tissue culture-fluorescent antibody method was satisfactory for demonstrating the virus in blood of infected animals 1 to 9 days postinfection, and in some organs after death. No false positive reactions were obtained.

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

    Directory of Open Access Journals (Sweden)

    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%.

  9. Immunofluorescence microscopy of paraffin-embedded human kidney specimens obtained by fine-needle aspiration biopsy.

    Science.gov (United States)

    Rantala, I; Laasonen, A; Pasternack, A; Mustonen, J

    1983-04-01

    Recently, we have introduced an atraumatic fine-needle aspiration biopsy method to obtain human glomeruli for morphologic investigation. In the present study, immunofluorescence microscopy of paraffin-embedded, fine-needle specimens is described. The specimens were obtained by aspiration with a 10-mL syringe fitted to the fine-needle prepared from a lumbar puncture needle (Jintan Terumo). Embedding of the specimens into conventional paraffin blocks was carried out after pelleting them by centrifugation between processing steps in conical centrifuge tubes. Sections from the blocks were collected on small pieces of GelBond film (FMC Corporation) instead of objective slides, which prevented the detachment of small sections during enzyme treatment. Localization then was performed on deparaffinized trypsin-digested sections using fluorescein-labeled antibodies. The choice of fixative and digestive enzymes was found to have a marked effect on the localization; periodate-lysine-paraformaldehyde fixative and trypsin digestion gave the most reliable results.

  10. Optimized detection of circulating anti-nuclear envelope autoantibodies by immunofluorescence

    Directory of Open Access Journals (Sweden)

    Kouroumalis Elias

    2006-09-01

    Full Text Available Abstract Background Antinuclear antibodies are useful diagnostic tools in several autoimmune diseases. However, the routine detection of nuclear envelope autoantibodies using immunofluorescence (IF is not always easy to perform in patients' sera because of the presence of autoantibodies to other nuclear and cytoplasmic components which could mask the characteristic rim-like pattern of nuclear envelope autoantibodies. This is particularly common in sera from patients with primary biliary cirrhosis (PBC, which generaly have high titres of anti-mitochondrial antibodies. Therefore, we have assayed a number of commercial slides and alternative fixation conditions to optimize the detection of anti-nuclear envelope antibodies (ANEA in PBC sera. Methods We have explored the presence of ANEA in 33 sera from patients with established PBC using three different Hep2 commercial slides and home-made slides with HeLa and Hep2 cells fixed with methanol, ethanol, 1% or 4% formaldehyde. Results We observed that the IF pattern was related to the cell type used (Hep2 or HeLa, the manufacturer and the cell fixation scheme. When both cell lines were fixed with 1% formaldehyde, the intensity of the cytoplasmic staining was considerably decreased regardless to the serum sample, whereas the prevalence of cytoplasmic autoantibodies was significantly lowered, as compared to any of the Hep2 commercial slide and fixation used. In addition, the prevalence of ANEA was importantly increased in formaldehyde-fixed cells. Conclusion Immunofluorescence using appropriately fixed cells represent an easy, no time-consuming and low cost technique for the routine screening of sera for ANEA. Detection of ANEA is shown to be more efficient using formaldehyde-fixed cells instead of commercially available Hep2 cells.

  11. 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.

  12. 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 Cav1.2 in an immunofluorescent approach. We performed immunocytochemical analysis of Cav1.2 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 Cav1.2 protein and a correct Cav1.2 expression pattern in polygonal shaped cells already 12h after transfection. A sequence of elaborated protocol modifications allows subcellular localization analysis of Cav1.2 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.

  13. The Importance of Zostera marina to a Local Food Web Based on the Analysis of Compound Specific Isotopes in Maquoit Bay, Gulf of Maine

    Science.gov (United States)

    Doolittle, H. A.; Johnson, B. J.; Ambrose, W. G.; Locke, W.; Harris, C. M.

    2010-12-01

    Zostera marina (also known as eelgrass) is an important primary producer in near shore ecosystems in the Gulf of Maine, providing both habitat and nutrients for a variety of organisms (e.g., crustaceans, polychaetes, gastropods, and fish). The purpose of this study is to use compound specific δ13C analyses of essential amino acids to determine the degree to which organic matter derived from isotopically distinct primary producers (e.g., eelgrass, phytoplankton, and epiphytic algae) contribute to the diets of snails, shrimp, and fish in an eelgrass system in Casco Bay. Maquoit Bay, located in northwestern Casco Bay, in the Gulf of Maine, is a shallow estuarine system that is characterized by silt and clay sized sediments and the presence of extensive eelgrass beds. Amino acid concentrations and δ13C compositions were determined for a variety of sample-types collected in July-August, 2010, from three sites in the study area, including muscle tissue from Tautogolabrus adspersus (cunner), Gasterosteus aculeatus (3-spined stickleback), Nassarius obsoletus (snail), and Mysis spp. (shrimp), seston (i.e., phytoplankton), Z. marina, and epiphytic algae. TFAA amino acid derivatives of the total hydrolyzate were analyzed by GC-FID for amino acid concentration, and by GC-c-IRMS- for carbon isotope composition. Muscle tissue was dominated by glutamic and aspartic acids, and leucine, whereas Zostera marina was dominated by aspartic and glutamic acids, and proline. Phenylalanine and leucine in Z. marina are approximately 10 ‰ enriched in 13C relative to these same amino acids in the seston. The carbon isotope values of these essential amino acids are significantly more enriched in 13C for N. obsoletus than for T. adspersus, G. aculeatus, and Mysis spp. These data suggest that organic matter derived from Z. marina and/or epiphytic algae is more important in the diets of N. obsoletus, and organic matter derived from seston is more important for the diets of T. adspersus, G

  14. Carbon sources in suspended particles and surface sediments from the Beaufort Sea revealed by molecular lipid biomarkers and compound-specific isotope analysis

    Directory of Open Access Journals (Sweden)

    I. Tolosa

    2013-03-01

    Full Text Available Molecular lipid biomarkers (hydrocarbons, alcohols, sterols and fatty acids and compound-specific isotope analysis of suspended particulate organic matter (SPM and surface sediments of the Mackenzie Shelf and slope (southeast Beaufort Sea, Arctic Ocean were studied in summer 2009. The concentrations of the molecular lipid markers, characteristic of known organic matter sources, were grouped and used as proxies to evaluate the relative importance of fresh algal, detrital algal, fossil, C3 terrestrial plants, bacterial and zooplankton material in the organic matter (OM of this area. Fossil and detrital algal contributions were the major fractions of the freshwater SPM from the Mackenzie River with ~34% each of the total molecular biomarkers. Fresh algal, C3 terrestrial, bacterial and zooplanktonic components represented much lower percentages, 17, 10, 4 and 80%, with a minor contribution of fossil and C3 terrestrial biomarkers. Characterization of the sediments revealed a major sink of refractory algal material mixed with some fresh algal material, fossil hydrocarbons and a small input of C3 terrestrial sources. In particular, the sediments from the shelf and at the mouth of the Amundsen Gulf presented the highest contribution of detrital algal material (60–75%, whereas those from the slope contained the highest proportion of fossil (40% and C3 terrestrial plant material (10%. Overall, considering that the detrital algal material is marine derived, autochthonous sources contributed more than allochthonous sources to the OM lipid pool. Using the ratio of an allochthonous biomarker (normalized to total organic carbon, TOC found in the sediments to those measured at the river mouth water, we estimated that the fraction of terrestrial material preserved in the sediments accounted for 30–40% of the total carbon in the inner shelf sediments, 17% in the outer shelf and Amundsen Gulf and up to 25% in the slope sediments. These estimates are low

  15. Distinguishing between old and modern permafrost sources in the northeast Siberian land-shelf system with compound-specific δ2H analysis

    Science.gov (United States)

    Vonk, Jorien E.; Tesi, Tommaso; Bröder, Lisa; Holmstrand, Henry; Hugelius, Gustaf; Andersson, August; Dudarev, Oleg; Semiletov, Igor; Gustafsson, Örjan

    2017-08-01

    Pleistocene ice complex permafrost deposits contain roughly a quarter of the organic carbon (OC) stored in permafrost (PF) terrain. When permafrost thaws, its OC is remobilized into the (aquatic) environment where it is available for degradation, transport or burial. Aquatic or coastal environments contain sedimentary reservoirs that can serve as archives of past climatic change. As permafrost thaw is increasing throughout the Arctic, these reservoirs are important locations to assess the fate of remobilized permafrost OC.We here present compound-specific deuterium (δ2H) analysis on leaf waxes as a tool to distinguish between OC released from thawing Pleistocene permafrost (ice complex deposits; ICD) and from thawing Holocene permafrost (from near-surface soils). Bulk geochemistry (%OC; δ13C; %total nitrogen, TN) was analyzed as well as the concentrations and δ2H signatures of long-chain n-alkanes (C21 to C33) and mid- to long-chain n-alkanoic acids (C16 to C30) extracted from both ICD-PF samples (n = 9) and modern vegetation and O-horizon (topsoil-PF) samples (n = 9) from across the northeast Siberian Arctic. Results show that these topsoil-PF samples have higher %OC, higher OC / TN values and more depleted δ13C-OC values than ICD-PF samples, suggesting that these former samples trace a fresher soil and/or vegetation source. Whereas the two investigated sources differ on the bulk geochemical level, they are, however, virtually indistinguishable when using leaf wax concentrations and ratios. However, on the molecular isotope level, leaf wax biomarker δ2H values are statistically different between topsoil PF and ICD PF. For example, the mean δ2H value of C29 n-alkane was -246 ± 13 ‰ (mean ± SD) for topsoil PF and -280 ± 12 ‰ for ICD PF. With a dynamic isotopic range (difference between two sources) of 34 to 50 ‰; the isotopic fingerprints of individual, abundant, biomarker molecules from leaf waxes can thus serve as endmembers to distinguish between

  16. Sources and transformation of dissolved and particulate organic nitrogen in the North Pacific Subtropical Gyre indicated by compound-specific δ15N analysis of amino acids

    Science.gov (United States)

    Yamaguchi, Yasuhiko T.; McCarthy, Matthew D.

    2018-01-01

    This study explores the use of compound-specific nitrogen isotopes of amino acids (δ15NAA) of coupled dissolved and particulate organic nitrogen (DON, PON) samples as a new approach to examine relative sources, transformation processes, and the potential coupling of these two major forms of N cycle in the ocean water column. We measured δ15NAA distributions in high-molecular-weight dissolved organic nitrogen (HMW DON) and suspended PON in the North Pacific Subtropical Gyre (NPSG) from surface to mesopelagic depths. A new analytical approach achieved far greater δ15NAA measurement precision for DON than earlier work, allowing us to resolve previously obscured differences in δ15NAA signatures, both with depth and between ON pools. We propose that δ15N values of total hydrolysable amino acids (THAA) represents a proxy for proteinaceous ON δ15N values in DON and PON. Together with bulk δ15N values, this allows δ15N values and changes in bulk, proteinaceous, and ;other-N; to be directly evaluated. These novel measurements suggest three main conclusions. First, the δ15NAA signatures of both surface and mesopelagic HMW DON suggest mainly heterotrophic bacterial sources, with mesopelagic HMW DON bearing signatures of far more degraded material compared to surface material. These results contrast with a previous proposal that HMW DON δ15NAA patterns are essentially ;pre-formed; by cyanobacteria in the surface ocean, undergo little change with depth. Second, different δ15NAA values and patterns of HMW DON vs. suspended PON in the surface NPSG suggest that sources and cycling of these two N reservoirs are surpisingly decoupled. Based on molecular δ15N signatures, we propose a new hypothesis that production of surface HMW DON is ultimately derived from subsurface nitrate, while PON in the mixed layer is strongly linked to N2 fixation and N recycling. In contrast, the comparative δ15NAA signatures of HMW DON vs. suspended PON in the mesopelagic also suggest a

  17. 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.

  18. Two languages, two sets of interpretations: Language-specific influences of morphological form on Dutch and English speakers' interpretation of compounds

    NARCIS (Netherlands)

    Banga, A.; Hanssen, E.J.M.; Schreuder, R.; Neijt, A.H.

    2013-01-01

    The present study investigates linguistic relativity. Do form differences between Dutch and English influence the interpretations which speakers have? The Dutch element en in noun-noun compounds, for example in aardbeienjam 'strawberry jam' is homophonous and homographic with the regular plural

  19. Complementing approaches to demonstrate chlorinated solvent biodegradation in a complex pollution plume: Mass balance, PCR and compound-specific stable isotope analysis.

    Science.gov (United States)

    Courbet, Christelle; Rivière, Agnès; Jeannottat, Simon; Rinaldi, Sandro; Hunkeler, Daniel; Bendjoudi, Hocine; de Marsily, Ghislain

    2011-11-01

    This work describes the use of different complementing methods (mass balance, polymerase chain reaction assays and compound-specific stable isotope analysis) to demonstrate the existence and effectiveness of biodegradation of chlorinated solvents in an alluvial aquifer. The solvent-contaminated site is an old chemical factory located in an alluvial plain in France. As most of the chlorinated contaminants currently found in the groundwater at this site were produced by local industries at various times in the past, it is not enough to analyze chlorinated solvent concentrations along a flow path to convincingly demonstrate biodegradation. Moreover, only a few data were initially available to characterize the geochemical conditions at this site, which were apparently complex at the source zone due to (i) the presence of a steady oxygen supply to the groundwater by irrigation canal losses and river infiltration and (ii) an alkaline pH higher than 10 due to former underground lime disposal. A demonstration of the existence of biodegradation processes was however required by the regulatory authority within a timeframe that did not allow a full geochemical characterization of such a complex site. Thus a combination of different fast methods was used to obtain a proof of the biodegradation occurrence. First, a mass balance analysis was performed which revealed the existence of a strong natural attenuation process (biodegradation, volatilization or dilution), despite the huge uncertainty on these calculations. Second, a good agreement was found between carbon isotopic measurements and PCR assays (based on 16S RNA gene sequences and functional genes), which clearly indicated reductive dechlorination of different hydrocarbons (Tetrachloroethene--PCE-, Trichloroethene--TCE-, 1,2-cisDichloroethene--cis-1,2-DCE-, 1,2-transDichloroethene-trans--1,2-DCE-, 1,1-Dichloroethene--1,1-DCE-, and Vinyl Chloride--VC) to ethene. According to these carbon isotope measurements, although TCE

  20. Resolving the bulk δ 15N values of ancient human and animal bone collagen via compound-specific nitrogen isotope analysis of constituent amino acids

    Science.gov (United States)

    Styring, Amy K.; Sealy, Judith C.; Evershed, Richard P.

    2010-01-01

    the biosynthetic pathway of threonine and the other amino acids. The δ 15N value of phenylalanine does not change significantly with trophic level, reflecting its conservative nature as an essential amino acid, and thus represents the isotopic composition of the nitrogen at the base of the food web. Δ 15N Glu-Phe values in particular are shown to reflect trophic level nitrogen sources within a food web. In relation to the reconstruction of ancient human diet the contribution of marine and terrestrial protein are strongly reflected in Δ 15N Glu-Phe values. Differences in nitrogen metabolism are also shown to have an influence upon individual amino acid δ 15N values with Δ 15N Glu-Phe values emphasising differences between the different physiological adaptations. The latter is demonstrated in tortoises, which can excrete nitrogen in the form of uric acid and urea and display negative Δ 15N Glu-Phe values whereas those for marine and terrestrial mammals are positive. The findings amplify the potential advantages of compound-specific nitrogen isotope analysis in the study of nitrogen flow within food webs and in the reconstruction of past human diets.

  1. Distinguishing between old and modern permafrost sources in the northeast Siberian land–shelf system with compound-specific δ2H analysis

    Directory of Open Access Journals (Sweden)

    J. E. Vonk

    2017-08-01

    Full Text Available Pleistocene ice complex permafrost deposits contain roughly a quarter of the organic carbon (OC stored in permafrost (PF terrain. When permafrost thaws, its OC is remobilized into the (aquatic environment where it is available for degradation, transport or burial. Aquatic or coastal environments contain sedimentary reservoirs that can serve as archives of past climatic change. As permafrost thaw is increasing throughout the Arctic, these reservoirs are important locations to assess the fate of remobilized permafrost OC.We here present compound-specific deuterium (δ2H analysis on leaf waxes as a tool to distinguish between OC released from thawing Pleistocene permafrost (ice complex deposits; ICD and from thawing Holocene permafrost (from near-surface soils. Bulk geochemistry (%OC; δ13C; %total nitrogen, TN was analyzed as well as the concentrations and δ2H signatures of long-chain n-alkanes (C21 to C33 and mid- to long-chain n-alkanoic acids (C16 to C30 extracted from both ICD-PF samples (n =  9 and modern vegetation and O-horizon (topsoil-PF samples (n =  9 from across the northeast Siberian Arctic. Results show that these topsoil-PF samples have higher %OC, higher OC ∕ TN values and more depleted δ13C-OC values than ICD-PF samples, suggesting that these former samples trace a fresher soil and/or vegetation source. Whereas the two investigated sources differ on the bulk geochemical level, they are, however, virtually indistinguishable when using leaf wax concentrations and ratios. However, on the molecular isotope level, leaf wax biomarker δ2H values are statistically different between topsoil PF and ICD PF. For example, the mean δ2H value of C29 n-alkane was −246 ± 13 ‰ (mean ± SD for topsoil PF and −280 ± 12 ‰ for ICD PF. With a dynamic isotopic range (difference between two sources of 34 to 50 ‰; the isotopic fingerprints of individual, abundant, biomarker molecules from leaf waxes can

  2. Medical Devices; Immunology and Microbiology Devices; Classification of the Automated Indirect Immunofluorescence Microscope and Software-Assisted System. Final order.

    Science.gov (United States)

    2017-11-14

    The Food and Drug Administration (FDA or we) is classifying the automated indirect immunofluorescence microscope and software-assisted system into class II (special controls). The special controls that apply to the device type are identified in this order and will be part of the codified language for the automated indirect immunofluorescence microscope and software-assisted system's classification. We are taking this action because we have determined that classifying the device into class II (special controls) will provide a reasonable assurance of safety and effectiveness of the device. We believe this action will also enhance patients' access to beneficial innovative devices, in part by reducing regulatory burdens.

  3. Improved Risk Analysis by Dual Direct Detection of Total and Infectious Cryptosporidium Oocysts on Cell Culture in Combination with Immunofluorescence Assay▿

    Science.gov (United States)

    Lalancette, Cindy; Di Giovanni, George D.; Prévost, Michèle

    2010-01-01

    The inactivation of Cryptosporidium oocysts is a main driver in the selection of water treatment disinfection strategies, and microbial risk analysis provides a sound basis for optimizing water treatment processes. U.S. Environmental Protection Agency method 1622/23 provides an estimate of the total oocyst count; however, it cannot be used directly for risk assessment, as it does not determine the fraction of infectious oocysts. Improved assessment of the risk for designated sources or in treated water requires evaluation of the total number of oocysts and an estimate of their infectivity. We developed a dual direct detection method using differential immunofluorescent staining that allows detection of both oocysts and cell culture infection foci for each sample. Using Cryptosporidium parvum oocysts, various pH levels, proteases, and gastroenteric compounds and substrates were assessed to determine their abilities to enhance the number of infection foci. The results showed that the key trigger for oocyst stimulation was acidification. Addition of a low concentration of d-glucose (50 mM) to the infection media increased rates of infectivity, while a higher dose (300 mM) was inhibitory. The total number of oocysts in each sample was determined by counting the oocysts remaining on a cell monolayer and the oocysts recovered from cell monolayer washes during processing using a simple filtration technique. With the dual direct detection on cell culture with immunofluorescence assay method, it is now possible to determine the numbers of total and infectious oocysts for a given sample in a single analysis. Direct percentages of infectivity are then calculated, which allows more accurate assessments of risk. PMID:19933339

  4. 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.

  5. Comparison of a rapid immunochromatographic assay with an immunofluorescent antibody test for detection of Leishmania infantum antibodies in dogs.

    Science.gov (United States)

    Proverbio, Daniela; Spada, Eva; Perego, Roberta; Baggiani, Luciana; Bagnagatti De Giorgi, Giada; Migliazzo, Antonella; Vitale, Fabrizio

    2016-12-01

    Identification of Leishmania infantum-infected dogs is crucial for control of canine leishmaniosis. In particular, in areas where access to specialized laboratories is limited, the availability of reliable and rapid in-clinic serologic tests may support immediate diagnosis in suspected cases and permit detection of asymptomatic canine carriers of L infantum infection. The purpose of the study was to validate the immunochromatographic test (ICT) Anigen Rapid Leishmania Ab Test kit for detection of L infantum antibodies in naturally exposed dogs in comparison with the immunofluorescence antibody test (IFAT). Serum samples from 66 dogs, including 20 healthy control dogs and 46 dogs suspected or confirmed with canine leishmaniosis, were measured by both tests. Anti-Leishmania IgG titers ≥ 1:40 by IFAT were considered positive. Kappa statistic with a 95% CI was calculated to evaluate agreement between the 2 testing methods, and sensitivity, specificity, and positive and negative likelihood ratio were calculated. Anti-L infantum IgG antibodies were found in 35 of 66 samples using the IFAT test (titers 1:40-1:5120). Thirty-one out of 66 samples tested positive with the qualitative ICT. Four IFAT-positive (titers ICT-negative. The Kappa value of 0.853 demonstrated very good agreement between the 2 tests. The Anigen Rapid Leishmania Ab Test kit reliably identified canine sera with anti-L infantum IgG antibody titers ≥ 1:40. The ICT requires neither special preparation of the serum nor specialized equipment and can be stored at ambient temperature. The test is applicable as a field test because it is easy to use and provides rapid results. © 2016 American Society for Veterinary Clinical Pathology.

  6. 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

    2017-11-29

    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.

  7. The Novel Anticytomegalovirus Compound AIC246 (Letermovir) Inhibits Human Cytomegalovirus Replication through a Specific Antiviral Mechanism That Involves the Viral Terminase ▿

    OpenAIRE

    Goldner, Thomas; Hewlett, Guy; Ettischer, Nicole; Ruebsamen-Schaeff, Helga; Zimmermann, Holger; Lischka, Peter

    2011-01-01

    Human cytomegalovirus (HCMV) remains the leading viral cause of birth defects and life-threatening disease in transplant recipients. All approved antiviral drugs target the viral DNA polymerase and are associated with severe toxicity issues and the emergence of drug resistance. Attempts to discover improved anti-HCMV drugs led to the identification of the small-molecular-weight compound AIC246 (Letermovir). AIC246 exhibits outstanding anti-HCMV activity in vitro and in vivo and currently is u...

  8. 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.

  9. In-vivo immunofluorescence confocal microscopy of herpes simplex virus type 1 keratitis

    Science.gov (United States)

    Kaufman, Stephen C.; Laird, Jeffery A.; Beuerman, Roger W.

    1996-05-01

    The white-light confocal microscope offers an in vivo, cellular-level resolution view of the cornea. This instrument has proven to be a valuable research and diagnostic tool for the study of infectious keratitis. In this study, we investigate the direct visualization of herpes simplex virus type 1 (HSV-1)-infected corneal epithelium, with in vivo confocal microscopy, using HSV-1 immunofluorescent antibodies. New Zealand white rabbits were infected with McKrae strain of HSV-1 in one eye; the other eye of each rabbit was used as an uninfected control. Four days later, the rabbits were anesthetized and a cellulose sponge was applied to each cornea, and a drop of direct HSV fluorescein-tagged antibody was placed on each sponge every 3 to 5 minutes for 1 hour. Fluorescence confocal microscopy was then performed. The HSV-infected corneas showed broad regions of hyperfluorescent epithelial cells. The uninfected corneas revealed no background fluorescence. Thus, using the confocal microscope with a fluorescent cube, we were able to visualize HSV-infected corneal epithelial cells tagged with a direct fluorescent antibody. This process may prove to be a useful clinical tool for the in vivo diagnosis of HSV keratitis.

  10. 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.

  11. 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.

  12. Is psoriasis an autoimmune disease? Interpretations from an immunofluorescence-based study.

    Science.gov (United States)

    Anand, Shashi; Gupta, Parikshaa; Bhardwaj, Ranjeet; Narang, Tarun; Dogra, Sunil; Minz, Ranjana W; Saikia, Biman; Chhabra, Seema

    2017-04-01

    Psoriasis is a multisystem, immune-mediated inflammatory disease. Some authors have proposed an autoimmune basis for psoriasis; however, till date, it has not been definitely established. This study was conducted to explore the autoimmune nature of psoriasis. This was a prospective study in which 43 psoriasis patients were assessed for detailed clinical, histopathological and immunopathological features to explore the diagnostic utility of subtypes, intensity and number of immunoreactants in lesional and non-lesional skin in these patients. In addition, the sera of these patients were analyzed for the presence of various autoantibodies. The patients' age ranged from 14 to 75 years with a male-to-female ratio of 1.52:1. Nine patients (20.93%) were positive for antinuclear and 2 (4.65%) for antismooth muscle antibodies. Direct immunofluorescence (DIF) was positive in 31 (72%) biopsies from the lesional and 27 (63%) biopsies from non-lesional skin. In all these DIF positive cases, granular deposits of C5b-9 were detected at the dermoepidermal junction. No significant difference was observed on comparing the type and pattern of immunoreactant positivity, among lesional and non-lesional skin biopsies (P > .05). No significant association between psoriasis and immunoreactant deposition as well as autoantibody seroprevalence was observed, thereby refuting a definite autoimmune basis for psoriasis. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  13. Immunofluorescence study of corneal wound healing after excimer laser anterior keratectomy in the monkey eye.

    Science.gov (United States)

    Malley, D S; Steinert, R F; Puliafito, C A; Dobi, E T

    1990-09-01

    We performed anterior keratectomies on six monkey eyes, four by excimer laser large-area ablation at 193 nm and two by mechanical keratectomy. Immunofluorescence was used to study the wound healing response histopathologically. The distribution of fibrinogen, fibronectin, laminin, collagen types III, IV, and VI, and keratan sulfate was determined at postoperative intervals of 24 hours, 6 days, and 1 month. At 24 hours, fibrinogen and fibronectin coated the ablated surface, but corneal epithelial cells had not yet migrated over the wound. By 6 days and persisting at 1 month, an epithelial ingrowth of seven to 10 layers, mild stromal hypercellularity, and new collagen formation were present in the repair region. At 1 month, fibrinogen, fibronectin, laminin, and type III collagen were strongly detected in the repair region. Type VI collagen was present in both normal and healed corneal stroma at all intervals, and type IV collagen was present in Descemet's membrane only. Sulfated keratan sulfate was absent from the newly synthesized collagen stroma at all intervals. Slit-lamp photographs demonstrated corneal haze in the ablation zone in all cases at 24 hours, persisting for 1 month. The fluorescence patterns produced by excimer laser ablation and mechanical keratectomy were qualitatively identical.

  14. Quantitative validation of immunofluorescence and lectin staining using reduced CLARITY acrylamide formulations.

    Science.gov (United States)

    Krolewski, D M; Kumar, V; Martin, B; Tomer, R; Deisseroth, K; Myers, R M; Schatzberg, A F; Lee, F S; Barchas, J D; Bunney, W E; Akil, H; Watson, S J

    2017-12-14

    The CLARITY technique enables three-dimensional visualization of fluorescent-labeled biomolecules in clarified intact brain samples, affording a unique view of molecular neuroanatomy and neurocircuitry. It is therefore, essential to find the ideal combination for clearing tissue and detecting the fluorescent-labeled signal. This method requires the formation of a formaldehyde-acrylamide fixative-generated hydrogel mesh through which cellular lipid is removed with sodium dodecyl sulfate. Several laboratories have used differential acrylamide and detergent concentrations to achieve better tissue clearing and antibody penetration, but the potential effects upon fluorescent signal retention is largely unknown. In an effort to optimize CLARITY processing procedures we performed quantitative parvalbumin immunofluorescence and lectin-based vasculature staining using either 4 or 8% sodium dodecyl sulfate detergent in combination with different acrylamide formulas in mouse brain slices. Using both confocal and CLARITY-optimized lightsheet microscope-acquired images, we demonstrate that 2% acrylamide monomer combined with 0.0125% bis-acrylamide and cleared with 4% sodium dodecyl sulfate generally provides the most optimal signal visualization amongst various hydrogel monomer concentrations, lipid removal times, and detergent concentrations.

  15. The Reliability of a Novel Automated System for ANA Immunofluorescence Analysis in Daily Clinical Practice

    Directory of Open Access Journals (Sweden)

    Mohammed Alsuwaidi

    2016-01-01

    Full Text Available Automated interpretation (AI systems for antinuclear antibody (ANA analysis have been introduced based on assessment of indirect immunofluorescence (IIF patterns. The diagnostic performance of a novel automated IIF reading system was compared with visual interpretation (VI of IIF in daily clinical practice to evaluate the reduction of workload. ANA-IIF tests of consecutive serum samples from patients with suspected connective tissue disease were carried out using HEp-2 cells according to routine clinical care. AI was performed using a visual analyser (Zenit G-Sight, Menarini, Germany. Agreement rates between ANA results by AI and VI were calculated. Of the 336 samples investigated, VI yielded 205 (61% negative, 42 (13% ambiguous, and 89 (26% positive results, whereas 82 (24% were determined to be negative, 176 (52% ambiguous, and 78 (24% positive by AI. AI displayed a diagnostic accuracy of 175/336 samples (52% with a kappa coefficient of 0.34 compared to VI being the gold standard. Solely relying on AI, with VI only performed for all ambiguous samples by AI, would have missed 1 of 89 (1% positive results by VI and misclassified 2 of 205 (1% negative results by VI as positive. The use of AI in daily clinical practice resulted only in a moderate reduction of the VI workload (82 of 336 samples: 24%.

  16. Magnetic, specific heat and electrical transport properties of Frank-Kasper cage compounds RTM2Al20 [R  =  Eu, Gd and La; TM  =  V, Ti

    Science.gov (United States)

    Kumar, K. Ramesh; Nair, Harikrishnan S.; Christian, Reinke; Thamizhavel, A.; Strydom, André M.

    2016-11-01

    Single crystals of Frank-Kasper compounds RTM2Al20 (R  =  Eu, Gd and La; TM  =  V and Ti) were grown by self-flux method and their physical properties were investigated through magnetization (M), magnetic susceptibility (χ), specific heat (C P) and electrical resistivity (ρ) measurements. Powder x-ray diffraction studies and structural analysis showed that these compounds crystallize in the cubic crystal structure with the space group Fd\\bar{3}m . The magnetic susceptibility for the compounds EuTi2Al20 and GdTi2Al20 showed a sudden jump below the Néel temperature T N indicative of plausible double magnetic transition. Specific heat (C P) and electrical resistivity (ρ) measurements also confirm the first-order magnetic transition (FOMT) and possible double magnetic transitions. Temperature variation of heat capacity showed a sharp phase transition and huge C P value for the (Eu/Gd)Ti2Al20 compounds’ full width at half-maximum (FWHM) (RTM2Al20 compounds. In contrast, linear variation of C P is observed in the ordered state for (Eu/Gd)V2Al20 compounds suggesting a λ-type transition. We observed clear anomaly between heating and cooling cycle in temperature-time relaxation curve for the compounds GdTi2Al20 (2.38 K) and EuTi2Al20 (3.2 K) which is indicating a thermal arrest due to the latent heat. The temperature variation of S mag for GdTi2Al20 saturates to a value 0.95R\\ln 8 while the other magnetic systems exhibited still lower entropy saturation values in the high temperature limit. ≤ft({{C}\\text{P}}-γ T\\right)/{{T}3} versus T plot showed a maximum near 27 K for all the compounds indicating the presence of low frequency Einstein modes of vibrations. Resistivity measurements showed that all the samples behave as normal Fermi liquid type compounds and ρ (T) due to electron-phonon scattering follows Bloch-Grüneisen-Mott relation in the paramagnetic region.

  17. Statistical evaluation of an improved quantitative immunofluorescence method of measuring serum antibody levels directed against intestinal bacteria

    NARCIS (Netherlands)

    Jansen, G.J.; Wilkinson, M.H.F.; Deddens, B.; Waaij, D. van der

    The accuracy of an improved quantitative immunofluorescence method to measure the antibody binding capacity of intestinal bacteria was assessed. The improvements comprise: calibration on a non-fading fluorescence standard (uranyl glass), prolonged camera exposure time (over 4 s) and shading

  18. 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

  19. OBJECTIVE QUANTITATION OF SERUM ANTIBODY-TITERS AGAINST ENTEROBACTERIACEAE USING INDIRECT IMMUNOFLUORESCENCE, READ BY VIDEOCAMERA AND IMAGE-PROCESSING SYSTEM

    NARCIS (Netherlands)

    APPERLOORENKEMA, HZ; WILKINSON, MHF; OENEMA, DG; VANDERWAAIJ, D

    1991-01-01

    A new way of measuring indirect immunofluorescence (IIF) of microscopic bacterial slide preparations by videocamera and an image processing system is presented. This method is compared with the conventional method of reading the slides by eye. The advantages of this new approach are objective

  20. Objective quantitation of serum antibody titres against Enterobacteriaceae using indirect immunofluorescence, read by videocamera and image processing system

    NARCIS (Netherlands)

    Apperloo-Renkema, H.Z.; Wilkinson, M.H.F.; Oenema, D.G.; Waaij, D. van der

    1991-01-01

    A new way of measuring indirect immunofluorescence (IIF) of microscopic bacterial slide preparations by videocamera and an image processing system is presented. This method is compared with the conventional method of reading the slides by eye. The advantages of this new approach are objective

  1. NOUN COMPOUND IN ENGLISH

    Directory of Open Access Journals (Sweden)

    Ni Luh Ketut Mas Indrawati

    2013-11-01

    Full Text Available This paper aims at analyzing and describing the English compound specifically the English noun compound. Compound is a combination of two or more words of which meaning cannot always be predicted from the meaning of each part. In English, words, especially adjectives and nouns, are combined into compound structures in a variety of ways. This article attempts to discuss the formal characteristics and types of the English noun compound. The theory of compound was adopted for further analysis. The finding shows that the formal characteristics of the English noun compound are: the noun compounds have primary stress on the first constituent, the semantic unity of a noun compound is reflected in an orthographic, the meaning of the noun compound cannot be predicted from the meaning of the parts. The orthographic characteristics can be solid, hyphenated, and open. The types involved are Subject and Verb, Verb and Object, verb and adverbial, verb-less, subject and complement, combining-form and Bahuvrihi

  2. Assessing the Origins of Aliphatic Amines in the Murchison Meteorite from their Compound-Specific Carbon Isotopic Ratios and Enantiomeric Composition

    Science.gov (United States)

    Aponte, Jose; Dworkin, Jason; Elsila, Jamie E.

    2014-01-01

    The study of meteoritic organic compounds provides a unique window into the chemical inventory of the early Solar System and prebiotic chemistry that may have been important for the origin of life on Earth. Multiple families of organic compounds have been extracted from the Murchison meteorite, which is one of the most thoroughly studied carbonaceous chondrites. The amino acids extracted from Murchison have been extensively analyzed, including measurements of non-terrestrial stable isotopic ratios and discoveries of L-enantiomeric excesses for alpha-dialkyl amino acids, notably isovaline. However, although the isotopic signatures of bulk amine-containing fractions have been measured, the isotopic ratios and enantiomeric composition of individual aliphatic amines, compounds that are chemically related to amino acids, remain unknown. Here, we report a novel method for the extraction, separation, identification and quantitation of aliphatic monoamines extracted from the Murchison meteorite. Our results show a complete suite of structural isomers, with a larger concentration of methylamine and ethylamine and decreasing amine concentrations with increasing carbon number. The carbon isotopic compositions of fourteen meteoritic aliphatic monoamines were measured, with delta C-13 values ranging from +21% to +129%, showing a decrease in C-13 with increasing carbon number, a relationship that may be consistent with the chain elongation mechanism under kinetic control previously proposed for meteoritic amino acids. We also found the enantiomeric composition of sec-butylamine, a structural analog to isovaline, was racemic within error, while the isovaline extracted from the same Murchison piece showed an L-enantiomeric excess of 9.7; this result suggested that processes leading to enantiomeric excess in the amino acid did not affect the amine. We used these collective data to assess the primordial synthetic origins of these meteoritic aliphatic amines and their potential

  3. Comparison of Simplexa Flu A/B & RSV PCR with Cytospin-Immunofluorescence and Laboratory-Developed TaqMan PCR in Predominantly Adult Hospitalized Patients

    Science.gov (United States)

    Ferguson, David

    2014-01-01

    To compare Simplexa Flu A/B & RSV PCR with cytospin-immunofluorescence and laboratory-developed TaqMan PCR methods, 445 nasopharyngeal samples were tested. Of these, 199 were positive (46 for respiratory syncytial virus [RSV], 120 for influenza A, and 33 for influenza B) and 246 were negative. The direct fluorescent-antibody assay (DFA) detected 132 (66.3%) positive samples, Simplexa direct detected 162 (81.4%), Simplexa using extracts detected 177 (88.9%), and lab-developed TaqMan PCR reference methods detected 199 (100%). The specificities were 99.6%, 100%, 100%, and 100%, respectively. The two Simplexa methods were more sensitive than the DFA (P = 0.0001) but less sensitive than the TaqMan reverse transcriptase PCR (P = 0.0001). PMID:24850350

  4. Development and Validation of a Specific Stability Indicating High Performance Liquid Chromatographic Methods for Related Compounds and Assay of Solifenacin Succinate

    Directory of Open Access Journals (Sweden)

    B. V. Rami Reddy

    2013-01-01

    Full Text Available Gradient, reverse phase liquid chromatographic methods were developed separately for the related compounds and solifenacin succinate, an active pharmaceutical ingredient used for the treatment of overactive bladder. Gradient LC method was employed for related compounds. The mobile phase-A contains a 0.01 M phosphate buffer pH: 3.5±0.05 with orthophosphoric acid (88% and mobile phase-B contains a mixture of acetonitrile and water in the ratio of 90 : 10(v/v. The flow rate was 1.0 mL/minute, column temperature was kept at 35°C, and detection was monitored at 220 nm. In the developed HPLC method the resolution between solifenacin succinate and its closely eluting impurity, that is, solifenacin N-oxide was found to be greater than 3.0. The drug was subjected to stress conditions such as hydrolysis, oxidation, photolysis, and thermal degradation. Considerable degradation was found to occur in only oxidative stress condition. Degradation product formed during oxidative stress condition was found to be impurity-C and it can be identified by LC-MS. The stress samples were assayed against a qualified reference standard and the mass balance was found close to 99.5%. The developed RP-LC method was validated as per ICH guidelines. We also developed LC-MS/MS method for determination and identification of these impurities in solifenacin succinate.

  5. Experimental bluetongue virus infection of sheep; effect of vaccination: pathologic, immunofluorescent, and ultrastructural studies.

    Science.gov (United States)

    Mahrt, C R; Osburn, B I

    1986-06-01

    Ten sheep were inoculated with bluetongue virus (BTV) serotype 17. Six of the sheep had been vaccinated before challenge exposure, 4 sheep served as nonvaccinated challenge-exposed controls, and 2 additional sheep served as nonvaccinated, nonchallenge-exposed, contact controls. Biopsy specimens (oral labial mucosa and skin) were obtained periodically after challenge exposure. Sheep were killed 8 to 13 days after challenge exposure, and necropsy was done. Vaccination did not seem to affect the nature or severity of the lesions observed. The changes in the mucosa of the cranial portion of the digestive tract included hyperemia, edema, inflammation, petechiae, erosions, ulcers, and surface encrustations. Lesions of skeletal, cardiac, and smooth muscles included hemorrhage, edema, myofiber degeneration, and necrosis. Lesions in cardiac muscles were sometimes widespread, indicating that cardiac failure may have been the major contributor to pulmonary congestion, edema, and eventual death during acute BTV infection. Damage to esophageal musculature resulted in vomiting. Hemorrhage was observed within the base of the pulmonary artery of all challenge-exposed sheep. Using immunofluorescence, bluetongue viral antigens were detected in small blood vessels of the skin, oral labial mucosa, tongue, esophagus, rumen, reticulum, urinary bladder, and pulmonary artery and in skeletal and cardiac muscles. Viral antigens were present in tissues obtained 3 to 11 days after inoculation. Ultrastructurally, changes in small-caliber blood vessels included congestion, hemorrhage, swollen degenerated endothelial cells, and occasional fibrin-platelet thrombi. Tubular structures and virus-like particles were observed within some of these endothelial cells.(ABSTRACT TRUNCATED AT 250 WORDS)

  6. Simplified CLARITY for visualizing immunofluorescence labeling in the developing rat brain.

    Science.gov (United States)

    Zheng, Huiyuan; Rinaman, Linda

    2016-05-01

    CLARITY is an innovative technological advance in which intact biological tissue is transformed into a "nanoporous hydrogel-hybridized form" (Chung et al. 2013; Chung and Deisseroth 2013) with markedly improved chemical and optical accessibility, permitting fluorescent visualization and extraction of high-resolution structural data from mm-thick blocks of tissue. CLARITY affords an excellent but as yet unexploited opportunity to visualize the growth and maturation of phenotypically identified neurons and axonal processes in the developing brain. This brief report describes a moderately revised, simplified, and less expensive CLARITY protocol that effectively reveals the structure of chemically identified neurons in whole neonatal/juvenile rat brains and tissue slabs. Rats [postnatal day (P)0-24] were transcardially perfused with one of two fixative/hydrogel solutions, followed by hydrogel polymerization to generate brain hybrids. Whole brain hybrids or 2.0-mm-thick coronal slabs were passively cleared of lipid and then processed for dual immunofluorescence labeling, including labeling using tyramide signal amplification. After refractive index matching using 2,20-Thiodiethanol (60 % solution), a Leica confocal microscope equipped with a CLARITY objective was used to view the hypothalamus in whole brain hybrids or slabs. Collected image stacks revealed the distribution and three-dimensional structure of hypothalamic pro-oxyphysin (oxytocin)-, neuropeptide Y-, glucagon-like peptide-1-, and tyrosine hydroxylase-immunopositive neurons and processes within large tissue volumes. Outstanding structural preservation and immunolabeling quality demonstrates the efficacy of this approach for interrogating chemically defined neural circuits as they develop in postnatal rodent brain.

  7. Compound-specific hydrogen isotope analysis of fluorine-, chlorine-, bromine- and iodine-bearing organics using gas chromatography-chromium-based high-temperature conversion (Cr/HTC) isotope ratio mass spectrometry.

    Science.gov (United States)

    Renpenning, Julian; Schimmelmann, Arndt; Gehre, Matthias

    2017-07-15

    The conventional high-temperature conversion (HTC) approach towards hydrogen compound-specific isotope analysis (CSIA) of halogen-bearing (F, Cl, Br, I) organics suffers from incomplete H2 yields and associated hydrogen isotope fractionation due to generation of HF, HCl, HBr, and HI byproducts. Moreover, the traditional off-line combustion of highly halogenated compounds results in incomplete recovery of water as an intermediary compound for hydrogen isotope ratio mass spectrometry (IRMS), and hence also leads to isotope fractionation. This study presents an optimized chromium-based high-temperature conversion (Cr/HTC) approach for hydrogen CSIA of various fluorinated, chlorinated, brominated and iodinated organic compounds. The Cr/HTC approach is fast, economical, and not affected by low H2 yields and associated isotope fractionation. The performance of the modified gas chromatography/chromium-based high-temperature conversion (GC-Cr/HTC) system was monitored and optimized using an ion trap mass spectrometer. Quantitative conversion of organic hydrogen into H2 analyte gas was achieved for all halogen-bearing compounds. The corresponding accuracy of CSIA was validated using (i) manual dual-inlet (DI)-IRMS after off-line conversion into H2 , and (ii) elemental analyzer (EA)-Cr/HTC-IRMS (on-line conversion). The overall hydrogen isotope analysis of F-, Cl-, Br- and I-bearing organics via GC-Cr/HTC-IRMS achieved a precision σ ≤ 3 mUr and an accuracy within ±5 mUr along the VSMOW-SLAP scale compared with the measured isotope compositions resulting from both validation methods, off-line and on-line. The same analytical performance as for single-compound GC-Cr/HTC-IRMS was achieved compound-specifically for mixtures of halogenated organics following GC separation to baseline resolution. GC-Cr/HTC technology can be implemented in existing analytical equipment using commercially available materials to provide a versatile tool for hydrogen CSIA of halogenated and non

  8. The Novel Anticytomegalovirus Compound AIC246 (Letermovir) Inhibits Human Cytomegalovirus Replication through a Specific Antiviral Mechanism That Involves the Viral Terminase ▿

    Science.gov (United States)

    Goldner, Thomas; Hewlett, Guy; Ettischer, Nicole; Ruebsamen-Schaeff, Helga; Zimmermann, Holger; Lischka, Peter

    2011-01-01

    Human cytomegalovirus (HCMV) remains the leading viral cause of birth defects and life-threatening disease in transplant recipients. All approved antiviral drugs target the viral DNA polymerase and are associated with severe toxicity issues and the emergence of drug resistance. Attempts to discover improved anti-HCMV drugs led to the identification of the small-molecular-weight compound AIC246 (Letermovir). AIC246 exhibits outstanding anti-HCMV activity in vitro and in vivo and currently is undergoing a clinical phase IIb trial. The initial mode-of-action studies suggested that the drug acts late in the HCMV replication cycle via a mechanism distinct from that of polymerase inhibitors. Here, we extend our mode-of-action analyses and report that AIC246 blocks viral replication without inhibiting the synthesis of progeny HCMV DNA or viral proteins. The genotyping of mutant viruses that escaped AIC246 inhibition uncovered distinct point mutations in the UL56 subunit of the viral terminase complex. Marker transfer analyses confirmed that these mutations were sufficient to mediate AIC246 resistance. The mapping of drug resistance to open reading frame UL56 suggests that viral DNA processing and/or packaging is targeted by AIC246. In line with this, we demonstrate that AIC246 affects the formation of proper unit-length genomes from viral DNA concatemers and interferes with virion maturation. However, since AIC246-resistant viruses do not exhibit cross-resistance to previously published terminase inhibitors, our data suggest that AIC246 interferes with HCMV DNA cleavage/packaging via a molecular mechanism that is distinct from that of other compound classes known to target the viral terminase. PMID:21752907

  9. The novel anticytomegalovirus compound AIC246 (Letermovir) inhibits human cytomegalovirus replication through a specific antiviral mechanism that involves the viral terminase.

    Science.gov (United States)

    Goldner, Thomas; Hewlett, Guy; Ettischer, Nicole; Ruebsamen-Schaeff, Helga; Zimmermann, Holger; Lischka, Peter

    2011-10-01

    Human cytomegalovirus (HCMV) remains the leading viral cause of birth defects and life-threatening disease in transplant recipients. All approved antiviral drugs target the viral DNA polymerase and are associated with severe toxicity issues and the emergence of drug resistance. Attempts to discover improved anti-HCMV drugs led to the identification of the small-molecular-weight compound AIC246 (Letermovir). AIC246 exhibits outstanding anti-HCMV activity in vitro and in vivo and currently is undergoing a clinical phase IIb trial. The initial mode-of-action studies suggested that the drug acts late in the HCMV replication cycle via a mechanism distinct from that of polymerase inhibitors. Here, we extend our mode-of-action analyses and report that AIC246 blocks viral replication without inhibiting the synthesis of progeny HCMV DNA or viral proteins. The genotyping of mutant viruses that escaped AIC246 inhibition uncovered distinct point mutations in the UL56 subunit of the viral terminase complex. Marker transfer analyses confirmed that these mutations were sufficient to mediate AIC246 resistance. The mapping of drug resistance to open reading frame UL56 suggests that viral DNA processing and/or packaging is targeted by AIC246. In line with this, we demonstrate that AIC246 affects the formation of proper unit-length genomes from viral DNA concatemers and interferes with virion maturation. However, since AIC246-resistant viruses do not exhibit cross-resistance to previously published terminase inhibitors, our data suggest that AIC246 interferes with HCMV DNA cleavage/packaging via a molecular mechanism that is distinct from that of other compound classes known to target the viral terminase.

  10. 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

  11. Immunofluorescence serration pattern analysis as a diagnostic criterion in antilaminin-332 mucous membrane pemphigoid: immunopathological findings and clinical experience in 10 Dutch patients.

    Science.gov (United States)

    Terra, J B; Pas, H H; Hertl, M; Dikkers, F G; Kamminga, N; Jonkman, M F

    2011-10-01

    Antilaminin-332 mucous membrane pemphigoid (anti-LN-332 MMP) is a chronic subepidermal blistering disease characterized by IgG anti-epidermal basement membrane zone (BMZ) autoantibodies against laminin-332 (LN-332). with anti-LN-332 MMP have an increased relative risk of malignancy. Laboratory techniques that are difficult to obtain are needed for diagnosis of anti-LN-332 MMP. Objectives  To incorporate direct immunofluorescence (DIF) serration pattern analysis of IgG depositions in the diagnostic criteria of anti-LN-332 MMP. Patients who met our revised inclusion criteria for anti-LN-332 MMP were selected from our biobank over the period 1997-2009. Inclusion criteria were clinical symptoms, DIF serration pattern analysis, indirect immunofluorescence (IIF) on salt-split skin, and antigen-specificity analysis of the serum including immunoblotting and/or immunoprecipitation and/or enzyme-linked immunosorbent assay (ELISA) against native LN-332. Ten patients met the inclusion criteria. A malignancy was found in two patients (20%). In all patients in whom it was performed (n = 9), DIF showed linear IgG deposition along the BMZ in an n-serrated pattern. Nine sera reacted by salt-split skin analysis and bound to the dermal side of the split skin. ELISA against native LN-332 was positive in 78% of the tested sera. Anti-LN-332 MMP can clinically resemble other forms of pemphigoid. Although state-of-the-art laboratory diagnostics are necessary for definite diagnosis, the combination of simple DIF serration pattern and IIF salt-split skin analysis will exclude other forms of MMP and epidermolysis bullosa acquisita from the differential diagnosis. Because of the increased risk for malignancy patients should be thoroughly oncologically screened. © 2011 The Authors. BJD © 2011 British Association of Dermatologists 2011.

  12. Comprehensive analytical methodology to determine hydrocarbons in marine waters using extraction disks coupled to glass fiber filters and compound-specific isotope analyses.

    Science.gov (United States)

    Ternon, Eva; Tolosa, Imma

    2015-07-24

    Solid-phase extraction of both aliphatic (AHs) and aromatic polycyclic hydrocarbons (PAHs) from seawater samples was evaluated using a GFF filter stacked upon an octadecyl bonded silica (C18) disk. Stable-isotope measurements were developed on hydrocarbons extracted from both GFF and C18-disks in order to characterize the source of hydrocarbons. A clear partition of hydrocarbon compounds between the dissolved and the particulate phase was highlighted. PAHs showed a higher affinity with the dissolved phase (recoveries efficiency of 48-71%) whereas AHs presented strong affinity with the particulate phase (up to 76% of extraction efficiency). Medium volumes of seawater samples were tested and no breakthrough was observed for a 5L sample. Isotopic fractionation was investigated within all analytical steps but none was evidenced. This method has been applied to harbor seawater samples and very low AH and PAH concentrations were achieved. Due to the low concentration levels of hydrocarbons in the samples, the source of hydrocarbons was determined by molecular indices rather than isotopic measurements and a pyrolytic origin was evidenced. The aliphatic profile also revealed the presence of long-chain linear alkylbenzenes (LABs). The methodology presented here would better fit to polluted coastal environments affected by recent oil spills. Copyright © 2015 Elsevier B.V. All rights reserved.

  13. Regioisomerism in the synthesis of a chiral aminotetralin drug compound: unraveling mechanistic details and diastereomer-specific in-depth NMR investigations.

    Science.gov (United States)

    Schuisky, Peter; Federsel, Hans-Jürgen; Tian, Wei

    2012-07-06

    During chemical process development of a novel 2-aminotetralin derivative intended for use as an antidepressant, scrutiny of the byproduct present in the drug molecule revealed a set of regioisomers. Detailed studies showed that this impurity issue originated from an early synthetic step in which a brominated tetralone motif was generated in a ring-closing protocol. It was found that this reaction was accompanied by a migration of the aromatic bromo substituent via different bromonium species along two discrete pathways. This example of the halogen dance reaction resulted in the formation of a series of tetralone impurities with a bromine distributed across all available aromatic positions of the tetralin nucleus. Subsequently, when subjected to reductive amination conditions, each of these tetralones gave rise to pairs of aminotetralins in a diastereomeric relationship. NMR investigations revealed that the alicyclic portion of the compounds thus formed displayed very complex signal patterns, which required further in-depth studies using a variety of sophisticated techniques. As a result, a deep insight into the structural features of the current 2-aminotetralin family was obtained, which is emphasized by the definition of a novel "0.2 ppm rule" allowing the absolute configuration at tetralin C-2 to be determined.

  14. Confocal and conventional immunofluorescent and immunogold electron microscopic localization of collagen types III and IV in human placenta.

    Science.gov (United States)

    Nanaev, A K; Rukosuev, V S; Shirinsky, V P; Milovanov, A P; Domogatsky, S P; Duance, V C; Bradbury, F M; Yarrow, P; Gardiner, L; d'Lacey, C

    1991-01-01

    Confocal and conventional indirect immunofluorescence and immunogold electron microscopic methods were applied to examine the distribution of extracellular matrix constituents (collagens types III and IV) in the villi of immature and term human placentae. The immunofluorescence study revealed that collagen type III is more distinct in the villous stroma of term placenta as compared with that of the first trimester. Collagen type IV was detected mainly in endothelial and epithelial basement membranes and interestingly also to a certain extent in the stroma. Results obtained using immunoelectron microscopy support the proposal that collagen types III and IV are characteristic of stromal and basement membranes, respectively. Stromal collagen type IV is apparently localized in association with the interstitial types of collagen (I and III), in the villous stroma of term placenta.

  15. Comparison of an avidin-biotin immunoassay with three commercially available immunofluorescence kits for typing of herpes simplex virus.

    OpenAIRE

    Barnard, D L; Johnson, F B; Richards, D F

    1985-01-01

    An avidin-biotin complex system was compared with three commercially available immunofluorescence kits for serotyping herpes simplex virus isolates from clinical specimens. Sensitivity values showed that the Electro-Nucleonics and Immulok reagents were useful in detecting the presence of virus, whereas the predictive values showed that the Syva and Immulok reagents possessed adequate discrimination between the herpes simplex virus serotypes. The avidin-biotin complex system was equal or super...

  16. New finding of Giardia intestinalis (Eukaryote, Metamonad in Old World archaeological site using immunofluorescence and enzyme-linked immunosorbent assays

    Directory of Open Access Journals (Sweden)

    Matthieu Le Bailly

    2008-05-01

    Full Text Available In this study, nine organic sediment samples from a medieval archaeological site at Pineuilh, France, were examined for Giardia intestinalis using two commercially available immunological kits [enzyme-linked immuno sorbent and immunofluorescence (IFA assays]. Both techniques detected G. intestinalis in one sample, dated to 1,000 Anno Domini. This is the first time IFA was successfully used to detect protozoa in Old World archaeological samples. Such immunological techniques offer important perspectives concerning ancient protozoa detection and identification.

  17. Self-assembly and sensing-group graft of pre-modified CNTs on resonant micro-cantilevers for specific detection of volatile organic compound vapors

    Science.gov (United States)

    Xu, Pengcheng; Li, Xinxin; Yu, Haitao; Liu, Min; Li, Jungang

    2010-11-01

    This paper reports MWCNT (multi-wall carbon nano-tube)-modified resonant micro-cantilever chemical sensors for detection of trinitrotoluene (TNT) vapor. The MWCNTs are pre-modified and then area-selectively self-assembled at the free-end gold pad of a micro-cantilever, in which a resonance-exciting heater and a signal-readout piezoresistive Wheatstone bridge are integrated. Featuring a high specific surface area, the MWCNTs are further functionalized with TNT-sensitive groups by grafting onto the sidewalls of the MWCNTs. To lower the non-specific absorption of water and other small organic molecules, the SiO2 surface of the micro-cantilever was also pre-treated for hydrophobicity and oleophobicity by self-assembling a monolayer of heptadecafluorodecyltrimethoxysilane. The results of our sensing experiments have shown a capability to rapidly detect ppb-level TNT vapor, and a high specificity of the functionalized groups to TNT molecules. The experiment has also confirmed a good long-term stability in detecting sensitivity.

  18. Lineage-specific SoxR-mediated Regulation of an Endoribonuclease Protects Non-enteric Bacteria from Redox-active Compounds.

    Science.gov (United States)

    Kim, Jisun; Park, Chulwoo; Imlay, James A; Park, Woojun

    2017-01-06

    Bacteria use redox-sensitive transcription factors to coordinate responses to redox stress. The [2Fe-2S] cluster-containing transcription factor SoxR is particularly tuned to protect cells against redox-active compounds (RACs). In enteric bacteria, SoxR is paired with a second transcription factor, SoxS, that activates downstream effectors. However, SoxS is absent in non-enteric bacteria, raising questions as to how SoxR functions. Here, we first show that SoxR of Acinetobacter oleivorans displayed similar activation profiles in response to RACs as did its homolog from Escherichia coli but controlled a different set of target genes, including sinE, which encodes an endoribonuclease. Expression, gel mobility shift, and mutational analyses indicated that sinE is a direct target of SoxR. Redox potentials and permeability of RACs determined optimal sinE induction. Bioinformatics suggested that only a few γ- and β-proteobacteria might have SoxR-regulated sinE Purified SinE, in the presence of Mg(2+) ions, degrades rRNAs, thus inhibiting protein synthesis. Similarly, pretreatment of cells with RACs demonstrated a role for SinE in promoting persistence in the presence of antibiotics that inhibit protein synthesis. Our data improve our understanding of the physiology of soil microorganisms by suggesting that both non-enteric SoxR and its target SinE play protective roles in the presence of RACs and antibiotics. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  19. Calibration of a Plant Uptake Model with Plant- and Site-Specific. Data for Uptake of Chlorinated Organic Compounds into Radish

    DEFF Research Database (Denmark)

    Trapp, Stefan

    2015-01-01

    The uptake of organic pollutants by plants is an important process for the exposure of humans to toxic chemicals. The objective of this study was to calibrate the parameters of a common plant uptake model by comparison to experimental results from literature. Radish was grown in contaminated soil...... with default data and site-specific data were similar. Deposition from air was the major uptake mechanism into shoots. Transport from soil with resuspended particles was only relevant for the contaminated plot. The calculation results (in dry weight) were most sensitive to changes of the water content of plant...

  20. Specific combination of compound heterozygous mutations in 17β-hydroxysteroid dehydrogenase type 4 (HSD17B4 defines a new subtype of D-bifunctional protein deficiency

    Directory of Open Access Journals (Sweden)

    McMillan Hugh J

    2012-11-01

    Full Text Available Abstract Background D-bifunctional protein (DBP deficiency is typically apparent within the first month of life with most infants demonstrating hypotonia, psychomotor delay and seizures. Few children survive beyond two years of age. Among patients with prolonged survival all demonstrate severe gross motor delay, absent language development, and severe hearing and visual impairment. DBP contains three catalytically active domains; an N-terminal dehydrogenase, a central hydratase and a C-terminal sterol carrier protein-2-like domain. Three subtypes of the disease are identified based upon the domain affected; DBP type I results from a combined deficiency of dehydrogenase and hydratase activity; DBP type II from isolated hydratase deficiency and DBP type III from isolated dehydrogenase deficiency. Here we report two brothers (16½ and 14 years old with DBP deficiency characterized by normal early childhood followed by sensorineural hearing loss, progressive cerebellar and sensory ataxia and subclinical retinitis pigmentosa. Methods and results Biochemical analysis revealed normal levels of plasma VLCFA, phytanic acid and pristanic acid, and normal bile acids in urine; based on these results no diagnosis was made. Exome analysis was performed using the Agilent SureSelect 50Mb All Exon Kit and the Illumina HiSeq 2000 next-generation-sequencing (NGS platform. Compound heterozygous mutations were identified by exome sequencing and confirmed by Sanger sequencing within the dehydrogenase domain (c.101C>T; p.Ala34Val and hydratase domain (c.1547T>C; p.Ile516Thr of the 17β-hydroxysteroid dehydrogenase type 4 gene (HSD17B4. These mutations have been previously reported in patients with severe-forms of DBP deficiency, however each mutation was reported in combination with another mutation affecting the same domain. Subsequent studies in fibroblasts revealed normal VLCFA levels, normal C26:0 but reduced pristanic acid beta-oxidation activity. Both DBP

  1. Immunofluorescence characterization of spinal cord dorsal horn microglia and astrocytes in horses

    Directory of Open Access Journals (Sweden)

    Constanza Stefania Meneses

    2017-10-01

    Full Text Available The role of glial cells in pain modulation has recently gathered attention. The objective of this study was to determine healthy spinal microglia and astrocyte morphology and disposition in equine spinal cord dorsal horns using Iba-1 and GFAP/Cx-43 immunofluorescence labeling, respectively. Five adult horses without visible wounds or gait alterations were selected. Spinal cord segments were obtained post-mortem for immunohistochemical and immunocolocalization assays. Immunodetection of spinal cord dorsal horn astrocytes was done using a polyclonal goat antibody raised against Glial Fibrillary Acidic Protein (GFAP and a polyclonal rabbit antibody against Connexin 43 (Cx-43. For immunodetection of spinal cord dorsal horn microglia, a polyclonal rabbit antibody against a synthetic peptide corresponding to the C-terminus of ionized calcium-binding adaptor molecule 1 (Iba-1 was used. Epifluorescence and confocal images were obtained for the morphological and organizational analysis. Evaluation of shape, area, cell diameter, cell process length and thickness was performed on dorsal horn microglia and astrocyte. Morphologically, an amoeboid spherical shape with a mean cell area of 92.4 + 34 µm2 (in lamina I, II and III was found in horse microglial cells, located primarily in laminae I, II and III. Astrocyte primary stem branches (and cellular bodies to a much lesser extent are mainly detected using GFAP. Thus, double GFAP/Cx-43 immunolabeling was needed in order to accurately characterize the morphology, dimension and cell density of astrocytes in horses. Horse and rodent astrocytes seem to have similar dimensions and localization. Horse astrocyte cells have an average diameter of 56 + 14 µm, with a main process length of 28 + 8 µm, and thickness of 1.4 + 0.3 µm, mainly situated in laminae I, II and III. Additionally, a close association between end-point astrocyte processes and microglial cell bodies was found. These results are the first

  2. Indirect immunofluorescence test in new world Leishmaniasis: serological and clinical relation-ship

    Directory of Open Access Journals (Sweden)

    Sergio C. F. Mendonça

    1988-09-01

    Full Text Available The indirect immunofluorescence test (IF for anti-Leishmania antibodies (IgC and IgM was performed with sera form the following groups of individuals: 214 cutaneous leishmaniasis patients, 28 healthy subjects with positive Montenegro's skin test (MST, 29 healthy subjects with negativeMST and 16 visceral leishmaniasis patients. The first four groups came from a suburban area of Rio de Janeiro (Jacarepaguá where cutaneous leishmaniasis caused by Leishmania braziliensis braziliensis is endemic. It was observed that IF-IgM titers were significantly higher amongst the cutaneous leishmaniasis patients with less than four months of disease as compared to those with longer periods and that IF-IgG titers were significantly higher in patients with multiple lesions as compared to those with single lesions. The visceral leishmaniasis patients had IF-IgG titers significantly higher than those from cutaneous leishmaniasis patients. A group of 28 individuals selected amongst the 214 cutaneous leishmaniasis patients had their IF-titers (IgG and IgM compared to those of the two control groups of healthy subjects from the endemic area, respectively with positive and negative MST. Significantly higher titers of IF-IgG and IF-IgM were found in the group with active disease. The same group of patients showed IF-IgG titers significantly lower at the end of the antimonial therapy than those observed during this tratment.O teste de imunofluorescência indireta (IF para a detecção de anticorpos anti-Leismania nas classes IgG e IgM foi realizado em soros de indivíduos dos seguintes grupos: 214 pacientes com leismaniose cutânea, quatro pacientes com leismaniose mucocutânea, 28 indivíduos sadios com intradermorreação de Montenegro (IDRM positiva, 29 indivíduos sadios com IDRM negativa e 16 pacientes com leishmaniose visceral. Os indivíduos dos quatro primeiros grupos eram provenientes de uma área da periferia da cidade do Rio de Janeiro (Jacarepaguá onde a

  3. Immunofluorescence characterization of spinal cord dorsal horn microglia and astrocytes in horses.

    Science.gov (United States)

    Meneses, Constanza Stefania; Müller, Heine Yacob; Herzberg, Daniel Eduardo; Uberti, Benjamín; Bustamante, Hedie Almagro; Werner, Marianne Patricia

    2017-01-01

    The role of glial cells in pain modulation has recently gathered attention. The objective of this study was to determine healthy spinal microglia and astrocyte morphology and disposition in equine spinal cord dorsal horns using Iba-1 and GFAP/Cx-43 immunofluorescence labeling, respectively. Five adult horses without visible wounds or gait alterations were selected. Spinal cord segments were obtained post-mortem for immunohistochemical and immunocolocalization assays. Immunodetection of spinal cord dorsal horn astrocytes was done using a polyclonal goat antibody raised against Glial Fibrillary Acidic Protein (GFAP) and a polyclonal rabbit antibody against Connexin 43 (Cx-43). For immunodetection of spinal cord dorsal horn microglia, a polyclonal rabbit antibody against a synthetic peptide corresponding to the C-terminus of ionized calcium-binding adaptor molecule 1 (Iba-1) was used. Epifluorescence and confocal images were obtained for the morphological and organizational analysis. Evaluation of shape, area, cell diameter, cell process length and thickness was performed on dorsal horn microglia and astrocyte. Morphologically, an amoeboid spherical shape with a mean cell area of 92.4 + 34 µm2 (in lamina I, II and III) was found in horse microglial cells, located primarily in laminae I, II and III. Astrocyte primary stem branches (and cellular bodies to a much lesser extent) are mainly detected using GFAP. Thus, double GFAP/Cx-43 immunolabeling was needed in order to accurately characterize the morphology, dimension and cell density of astrocytes in horses. Horse and rodent astrocytes seem to have similar dimensions and localization. Horse astrocyte cells have an average diameter of 56 + 14 µm, with a main process length of 28 + 8 µm, and thickness of 1.4 + 0.3 µm, mainly situated in laminae I, II and III. Additionally, a close association between end-point astrocyte processes and microglial cell bodies was found. These results are the first

  4. Neuron-Specific Fluorescence Reporter-Based Live Cell Tracing for Transdifferentiation of Mesenchymal Stem Cells into Neurons by Chemical Compound.

    Science.gov (United States)

    Hwang, Do Won; Kwon, Hyun Woo; Jang, Jaeho; Jung, Hee Jung; Kim, Kwang Rok; Lee, Dong Soo

    2017-01-01

    Although transdifferentiation of mesenchymal stem cells (MSCs) into neurons increases the possibility of therapeutic use of MSCs for neurodevelopmental disorders, the use of MSCs has the limitation on differentiation efficiency to neuronal lineage and lack of an easy method to monitor the transdifferentiation. In this study, using time-lapse live cell imaging, we assessed the neuronal differentiation of MSCs induced by a small molecule "NHPDQC (N-hydroxy-2-oxo-3-(3-phenylprophyl)-1,2-dihydroquinoxaline-6-carboxamide, C18H17N3O3)." Plasmid vector containing red fluorescence reporter genes under the control of the tubulin α1 (Tα1) promoter (pTα1-DsRed2) traced the neuronal differentiation of MSCs. Two days after NHPDQC treatment, MSCs showed neuron-like phenotype with neurite outgrowth and high expression of neuron-specific markers in more than 95% cells. The fluorescence signals increased in the cytoplasm of pTα1-DsRed2-transfected MSCs after NHPDQC treatment. In vitro monitoring of MSCs along the time courses showed progressive increase of fluorescence till 30 h after treatment, corresponding with the increase in neurite length. We examined an efficient neuronal differentiation of MSCs by NHPDQC alone and monitored the temporal changes of neuronal differentiation by neuron-specific fluorescence reporter along time. This method would help further our understanding of the differentiation of MSCs to produce neurons by simple treatment of small molecule.

  5. Compound- and sex-specific effects on programming of energy and immune homeostasis in adult C57BL/6JxFVB mice after perinatal TCDD and PCB 153.

    Science.gov (United States)

    van Esterik, J C J; Verharen, H W; Hodemaekers, H M; Gremmer, E R; Nagarajah, B; Kamstra, J H; Dollé, M E T; Legler, J; van der Ven, L T M

    2015-12-01

    Early life exposure to endocrine disrupting compounds has been linked to chronic diseases later in life, like obesity and related metabolic disorders. We exposed C57BL/6JxFVB hybrid mice to the aryl hydrocarbon receptor agonist 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and the constitutive androstane receptor/pregnane X receptor agonist polychlorinated biphenyl 153 (PCB 153) in an experimental design relevant for human exposure. Exposure occurred during gestation and lactation via maternal feed to a wide dose range (TCDD: 10-10,000 pg/kg body weight/day; PCB 153: 0.09-1406 μg/kg body weight/d). Then exposure was ceased and offspring were followed up to 1 year of age. Metabolic parameters like body weight, fat pad weights, glucose tolerance, endocrine serum profile, and neurobehavioral and immunological parameters were determined. Body weight was transiently affected by both compounds throughout the follow-up. TCDD-exposed males showed decreased fat pad and spleen weights and an increase in IL-4 production of splenic immune cells. In contrast, females showed increased fat pad weights and production of IFNγ. PCB 153-exposed males showed an increase in glucose, whereas females showed an increase in glucagon, a decrease in pancreas weight, and an increase in thymus weight. In conclusion, early life exposure to TCDD appears to affect programming of energy and immune homeostasis in offspring, whereas the effects of perinatal PCB 153 were mainly on programming of glucose homeostasis. Both compounds act sex-specifically. Lowest derived BMDLs (lower bounds of the (two sided) 90%-confidence interval for the benchmark dose) for both compounds are not lower than current tolerable daily intakes.

  6. The coordinated p53 and estrogen receptor cis-regulation at an FLT1 promoter SNP is specific to genotoxic stress and estrogenic compound.

    Directory of Open Access Journals (Sweden)

    Yari Ciribilli

    Full Text Available BACKGROUND: Recently, we established that a C>T single nucleotide polymorphism (SNP in the promoter of the VEGF receptor FLT1 gene generates a (1/2 site p53 response element (RE-T that results in p53 responsiveness of the promoter. The transcriptional control required an estrogen receptor (ER (1/2 site response element (ERE1 225 nt upstream to the RE-T. METHODOLOGY/PRINCIPAL FINDINGS: Here we report the identification of a second ER (1/2 site (ERE2 located 145 bp downstream of the RE-T and establish that both EREs can impact p53-mediated transactivation of FLT1-T in a manner that is cell type and ER level dependent. Gene reporter assays and ChIP experiments conducted in the breast cancer-derived MCF7 cells revealed that the ERE2 site was sufficient for p53-mediated ERalpha recruitment and transactivation of the FLT1-T promoter/reporter construct. Surprisingly, unlike the case for other p53 target promoters, p53-mediated transactivation of FLT1-T constructs or expression of the endogenous FLT1 gene, as well as binding of p53 and ER at the promoter constructs, was inducible by doxorubicin but not by 5-fluorouracil. Furthermore, ER activity at FLT1-T was differentially affected by ER ligands, compared to a control TFF1/pS2 ER target promoter. The p53-related transcription factors (TFs p73 and p63 had no effect on FLT1 transactivation. CONCLUSIONS/SIGNIFICANCE: We establish a new dimension to the p53 master regulatory network where p53-mediated transcription from a (1/2 site RE can be determined by ER binding at one or more cis-acting EREs in manner that is dependent on level of ER protein, the type of ER ligand and the specific p53-inducing agent.

  7. Promiscuity progression of bioactive compounds over time

    OpenAIRE

    Hu, Ye; Jasial, Swarit; Bajorath, J?rgen

    2015-01-01

    In the context of polypharmacology, compound promiscuity is rationalized as the ability of small molecules to specifically interact with multiple targets. To study promiscuity progression of bioactive compounds in detail, nearly 1 million compounds and more than 5.2 million activity records were analyzed. Compound sets were assembled by applying different data confidence criteria and selecting compounds with activity histories over many years. On the basis of release dates, compounds and acti...

  8. Multipurpose Compound

    Science.gov (United States)

    1983-01-01

    Specially formulated derivatives of an unusual basic compound known as Alcide may be the answer to effective treatment and prevention of the disease bovine mastitis, a bacterial inflammation of a cow's mammary gland that results in loss of milk production and in extreme cases, death. Manufactured by Alcide Corporation the Alcide compound has killed all tested bacteria, virus and fungi, shortly after contact, with minimal toxic effects on humans or animals. Alcide Corporation credits the existence of the mastitis treatment/prevention products to assistance provided the company by NERAC, Inc.

  9. cGMP-Elevating Compounds and Ischemic Conditioning Provide Cardioprotection Against Ischemia and Reperfusion Injury via Cardiomyocyte-Specific BK Channels.

    Science.gov (United States)

    Frankenreiter, Sandra; Bednarczyk, Piotr; Kniess, Angelina; Bork, Nadja I; Straubinger, Julia; Koprowski, Piotr; Wrzosek, Antoni; Mohr, Eva; Logan, Angela; Murphy, Michael P; Gawaz, Meinrad; Krieg, Thomas; Szewczyk, Adam; Nikolaev, Viacheslav O; Ruth, Peter; Lukowski, Robert

    2017-12-12

    The nitric oxide-sensitive guanylyl cyclase/cGMP-dependent protein kinase type I signaling pathway can afford protection against the ischemia/reperfusion injury that occurs during myocardial infarction. Reportedly, voltage and Ca2+-activated K+ channels of the BK type are stimulated by cGMP/cGMP-dependent protein kinase type I, and recent ex vivo studies implicated that increased BK activity favors the survival of the myocardium at ischemia/reperfusion. It remains unclear, however, whether the molecular events downstream of cGMP involve BK channels present in cardiomyocytes or in other cardiac cell types. Gene-targeted mice with a cardiomyocyte- or smooth muscle cell-specific deletion of the BK (CMBK or SMBK knockouts) were subjected to the open-chest model of myocardial infarction. Infarct sizes of the conditional mutants were compared with litter-matched controls, global BK knockout, and wild-type mice. Cardiac damage was assessed after mechanical conditioning or pharmacological stimulation of the cGMP pathway and by using direct modulators of BK. Long-term outcome was studied with respect to heart functions and cardiac fibrosis in a chronic myocardial infarction model. Global BK knockouts and CMBK knockouts, in contrast with SMBK knockouts, exhibited significantly larger infarct sizes compared with their respective controls. Ablation of CMBK resulted in higher serum levels of cardiac troponin I and elevated amounts of reactive oxygen species, lower phosphorylated extracellular receptor kinase and phosphorylated AKT levels and an increase in myocardial apoptosis. Moreover, CMBK was required to allow beneficial effects of both nitric oxide-sensitive guanylyl cyclase activation and inhibition of the cGMP-degrading phosphodiesterase-5, ischemic preconditioning, and postconditioning regimens. To this end, after 4 weeks of reperfusion, fibrotic tissue increased and myocardial strain echocardiography was significantly compromised in CMBK-deficient mice. Lack of CMBK

  10. Comparison of antibody capture radio- and enzyme immunoassays with immunofluorescence for detecting IgM antibody in infants with congenital rubella

    Energy Technology Data Exchange (ETDEWEB)

    Chantler, S.; Evans, C.J. (Wellcome Research Lab., Beckenham (UK)); Mortimer, P.P. (Virus Reference Laboratory, Central Public Health Laboratory, Colindale Avenue, London, (UK)); Cradock-Watson, J.E.; Ridehalgh, M.K.S. (Withington Hospital, Manchester (UK))

    1982-08-01

    IgM antibody capture radioimmunoassay (MACRIA) and enzyme immunoassay (MACEIA) were compared with immunofluorescence (IF) for detecting specific IgM antibody in 99 sera from 76 infants with confirmed congenital rubella, and 61 sera from a comparative group of 59 infants who had miscellaneous abnormalities but in whom congenital rubella was not confirmed. All of 35 specimens collected from confirmed cases within 12 weeks of birth were positive by all three methods and all but one of 17 specimens collected after the age of 18 months were uniformly negative. At intermediate ages discrepancies occurred in 18 specimens, of which eight were positive and 10 negative by IF. Three of these 18 specimens were negative by both antibody capture procedures but showed weak fluorescence; the other 15 were negative by MACEIA, but positive by MACRIA which appears to be the more sensitive of the antibody capture methods. Sera from five infants in the comparative group were clearly positive by all three methods. These five infants were probably congenitally infected with rubella. Sera from the other 54 infants were negative, except for one that gave a weakly positive result by MACRIA alone. Antibody capture procedures offer several advantages over previous methods for detecting IgM antibody. Although MACRIA was found to be slightly more sensitive than MACEIA, the greater stability of the enzyme label, together with the possibility of both visual and quantitative assessment, could make MACEIA the method of choice for detecting rubella-specific IgM.

  11. Detection of herpes simplex virus-related antigens in the nuclei and cytoplasm of biochemically transformed cells with peroxidase/anti-peroxidase immunological staining and indirect immunofluorescence.

    Science.gov (United States)

    Kurchak, M; Dubbs, D R; Kit, S

    1977-09-15

    Herpes simplex virus (HSV)-related antigens have been demonstrated in the nuclei and cytoplasm of human and mouse cells biochemically transformed by ultraviolet light-irradiated HSV. This was accomplished by using peroxidase/anti-peroxidase immunological staining and indirect immunofluorescence with rabbit antisera that had high neutralizing titers against the HSV-specific thymidine kinase activity and virus infectivity. HSV-1 antisera reacted with antigens in cells biochemically transformed by type 1 HSV, but not with those of cells biochemically transformed by type 2 HSV. Similarly, HSV-2 antisera reacted with antigens in cells biochemically transformed by HSV-2, but not with those in cells biochemically transformed by HSV-1. In contrast, herpes virus-related antigens were detected in cells cytolytically infected with HSV-1 and with HSV-2 by either type 1 or type 2 HSV antisera. These observations suggest that the antigens detected in the biochemically transformed cells were a type-specific subset of the HSV-related antigens synthesized in cells undergoing productive infection by HSV-1 and HSV-2.

  12. Mesoionic Compounds

    Indian Academy of Sciences (India)

    Home; Journals; Resonance – Journal of Science Education; Volume 11; Issue 10. Mesoionic Compounds - An Unconventional Class of Aromatic Heterocycles. Bharati V Badami. General Article Volume 11 Issue 10 October 2006 pp 40-48. Fulltext. Click here to view fulltext PDF. Permanent link:

  13. Mesoionic Compounds

    Indian Academy of Sciences (India)

    Sydnone, the representative mesoionic compound has been extensively studied because of its unusual structure, chemi- cal properties and synthetic utility. Sydnone is used as a versatile synthon in heterocyclic synthesis. This article gives a brief account of the comparative studies of the structural features of mesoionic ...

  14. Immunofluorescent localization of collagen types I, III, IV, V, fibronectin, laminin, entactin, and heparan sulphate proteoglycan in human immature placenta.

    Science.gov (United States)

    Rukosuev, V S

    1992-03-15

    The distribution of eight components of the extracellular matrix in immature human placenta was studied by an indirect immunofluorescence method with monospecific antibodies. In the stroma of the term chorionic villi, collagen types I, III, IV, V, and fibronectin formed a mesh of fibers and conglomerates. Heparan sulphate proteoglycan formed multiple conglomerates, whereas laminin comprised small, scanty, discrete granules. Collagen type IV, laminin, entactin, and heparan sulphate proteoglycan were confined to the basement membrane of the trophoblast. Sometimes, only collagen type IV was identified in fetal vascular basement membrane.

  15. COMPARISON OF INDIRECT IMMUNOFLUORESCENCE WITH AGAR-GEL IMMUNODIFFUSION FOR THE DIAGNOSIS OF BLUETONGUE VIRUS-INFECTION

    OpenAIRE

    ARITA, GMM; GATTI, MSV; GERMANO, PML; PESTANADECASTRO, AF

    1992-01-01

    1. Sera from 190 cows and from 72 sheep were examined to compare the results obtained with the agar gel immunodiffusion (AGID) and indirect immunofluorescence (IIF) tests for the diagnosis of bluetongue (BT) disease. 2. In the AGID test, 96 of 190 (50.5%) cattle serum samples and 38 of 72 (52.7%) sheep serum samples were positive, for a total of 134 out of 262 (51.1%) sera. In the IIF test, 98 of 190 (51.6%) cattle serum samples and 39 of 72 (54.2%) sheep serum samples were positive, for a to...

  16. An assessment of the immunofluorescence technique as a method for demonstrating the histological localization of tetrahydrocannabinol in mammalian tissues

    Energy Technology Data Exchange (ETDEWEB)

    Morley, M.; Gee, D.J.

    1982-10-01

    The use of the indirect immunofluorescence technique as a method for demonstrating the histological localization of tetrahydrocannabinol (delta-THC) has been examined. The experimental protocol was designed in order that optimal staining conditions with respect to temperature, the length of time of incubations and washes, and the dilution of the antisera should be defined. No marked differences were detected between frozen sections of liver from normal and delta-THC-injected mice. Results from radiotracer experiments using human liver suggest that the success of the method is dependent upon the solubility characteristics of the antigen-antibody complex.

  17. 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 HEp-2 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...

  18. Indirect immunofluorescence colony staining method for detecting bacterial pathogens of tomato

    NARCIS (Netherlands)

    Veena, M.S.; Vuurde, van J.W.L.

    2002-01-01

    An indirect immunoflouorescence colony staining method was developed for the detection of important seed-borne bacterial pathogens of tomato. The method involves the use of specific antiserum for initial binding of target bacteria and visualization of positive colonies with a commercially available

  19. Longitudinal imaging of HIV-1 spread in humanized mice with parallel 3D immunofluorescence and electron tomography.

    Science.gov (United States)

    Kieffer, Collin; Ladinsky, Mark S; Ninh, Allen; Galimidi, Rachel P; Bjorkman, Pamela J

    2017-02-15

    Dissemination of HIV-1 throughout lymphoid tissues leads to systemic virus spread following infection. We combined tissue clearing, 3D-immunofluorescence, and electron tomography (ET) to longitudinally assess early HIV-1 spread in lymphoid tissues in humanized mice. Immunofluorescence revealed peak infection density in gut at 10-12 days post-infection when blood viral loads were low. Human CD4+ T-cells and HIV-1-infected cells localized predominantly to crypts and the lower third of intestinal villi. Free virions and infected cells were not readily detectable by ET at 5-days post-infection, whereas HIV-1-infected cells surrounded by pools of free virions were present in ~10% of intestinal crypts by 10-12 days. ET of spleen revealed thousands of virions released by individual cells and discreet cytoplasmic densities near sites of prolific virus production. These studies highlight the importance of multiscale imaging of HIV-1-infected tissues and are adaptable to other animal models and human patient samples.

  20. Temporal appearance of structural and nonstructural bluetongue viral proteins in infected cells, as determined by immunofluorescence staining and flow cytometry.

    Science.gov (United States)

    Whetter, L E; Gebhard, D H; MacLachlan, N J

    1990-08-01

    The temporal appearance of 4 viral proteins was detected in bluetongue virus (BTV)-infected Vero cells by indirect immunofluorescence staining with monoclonal antibodies (MAb) to BTV structural proteins VP2 and VP7 and nonstructural proteins NS1 and NS2. Bluetongue viral proteins were detected at distinct intervals after inoculation of Vero cells; VP7 was first detected 3 hours after inoculation, NS1 and NS2 at 5 hours after inoculation, whereas VP2 was not detected until 8 hours after inoculation. Patterns of fluorescence varied with the fixative used, but each MAb induced a distinct pattern of fluorescence in infected cells. Flow cytometry, which was used with each of the 4 MAb, proved to be an accurate and sensitive method of detecting BTV-infected P3 mouse myeloma cells. The temporal appearance of each viral protein in BTV-infected P3 cells was similar to that detected in BTV-infected Vero cells. Advantages of flow cytometry over conventional immunofluorescence staining to detect BTV-infected cells included: (1) enumeration of the proportion of infected cells in a population; (2) further characterization of infected cells, including estimates of their viability; and (3) computer-assisted storage and analysis of data obtained.

  1. Preparation of a bio-immunoreagent between ZZ affibody and enhanced green fluorescent protein for immunofluorescence applications.

    Science.gov (United States)

    Yang, Hong-Ming; Chen, Yong; Gao, Zhi-Qin; Tang, Jin-Bao

    2012-03-01

    In the present study, we constructed plasmid pUC-ZZ-EGFP to express Pro-ZZ-EGFP using ZZ peptide (a synthetic artificial IgG-Fc-fragment-binding protein derived from the B domain of staphylococcal protein A) and enhanced green fluorescent protein (EGFP). Without induction with isopropyl-β-D: -thiogalactopyranoside, the chimeric protein was effectively expressed in Escherichia coli HB101. Its affinity constant binding IgG was 2.6 × 10(8) M(-1) obtained by competitive enzyme-linked immunosorbent assay, indicating that the ZZ peptide retains the native structure in Pro-ZZ-EGFP. The application of immunofluorescence assay for detecting the Mycoplasma pneumoniae IgG antibody, Pro-ZZ-EGFP, exhibited a good signal comparable in brightness and fluorescence pattern with the signal generated using the fluorescein isothiocyanate-labeled anti-human IgG. The result indicates that Pro-ZZ-EGFP possesses great potential for clinical immunofluorescence IgG test as an alternative versatile fluorescent antibody.

  2. Compound odontoma

    Directory of Open Access Journals (Sweden)

    Monica Yadav

    2012-01-01

    Full Text Available Odontomas have been extensively reported in the dental literature, and the term refers to tumors of odontogenic origin. Though the exact etiology is still unknown, the postulated causes include: local trauma, infection, inheritance and genetic mutation. The majority of the lesions are asymptomatic; however, may be accompanied with pain and swelling as secondary complaints in some cases. Here, we report a case of a compound odontome in a 14 year old patient.

  3. Compound-specific δ15N amino acid measurements in littoral mussels in the California upwelling ecosystem: a new approach to generating baseline δ15N Isoscapes for coastal ecosystems.

    Directory of Open Access Journals (Sweden)

    Natasha L Vokhshoori

    Full Text Available We explored δ(15N compound-specific amino acid isotope data (CSI-AA in filter-feeding intertidal mussels (Mytilus californianus as a new approach to construct integrated isoscapes of coastal primary production. We examined spatial δ(15N gradients in the California Upwelling Ecosystem (CUE, determining bulk δ(15N values of mussel tissue from 28 sites between Port Orford, Oregon and La Jolla, California, and applying CSI-AA at selected sites to decouple trophic effects from isotopic values at the base of the food web. Bulk δ(15N values showed a strong linear trend with latitude, increasing from North to South (from ∼ 7‰ to ∼ 12‰, R(2 = 0.759. In contrast, CSI-AA trophic position estimates showed no correlation with latitude. The δ(15N trend is therefore most consistent with a baseline δ(15N gradient, likely due to the mixing of two source waters: low δ(15N nitrate from the southward flowing surface California Current, and the northward transport of the California Undercurrent (CUC, with (15N-enriched nitrate. This interpretation is strongly supported by a similar linear gradient in δ(15N values of phenylalanine (δ(15NPhe, the best AA proxy for baseline δ(15N values. We hypothesize δ(15N(Phe values in intertidal mussels can approximate annual integrated δ(15N values of coastal phytoplankton primary production. We therefore used δ(15N(Phe values to generate the first compound-specific nitrogen isoscape for the coastal Northeast Pacific, which indicates a remarkably linear gradient in coastal primary production δ(15N values. We propose that δ(15N(Phe isoscapes derived from filter feeders can directly characterize baseline δ(15N values across major biochemical provinces, with potential applications for understanding migratory and feeding patterns of top predators, monitoring effects of climate change, and study of paleo- archives.

  4. Evaluation of quantum dot conjugated antibodies for immunofluorescent labelling of cellular targets

    Directory of Open Access Journals (Sweden)

    Jennifer E. Francis

    2017-06-01

    Full Text Available Semiconductor quantum dots (Qdots have been utilised as probes in fluorescence microscopy and provide an alternative to fluorescent dyes and fluorescent proteins due to their brightness, photostability, and the possibility to excite different Qdots with a single wavelength. In spite of these attractive properties, their implemenation by biologists has been somewhat limited and only a few Qdot conjugates are commercially available for the labelling of cellular targets. Although many protocols have been reported for the specific labelling of proteins with Qdots, the majority of these relied on Qdot-conjugated antibodies synthesised specifically by the authors (and therefore not widely available, which limits the scope of applications and complicates replication. Here, the specificity of a commercially available, Qdot-conjugated secondary antibody (Qdot-Ab was tested against several primary IgG antibodies. The antigens were labelled simultaneously with a fluorescent dye coupled to a secondary antibody (Dye-Ab and the Qdot-Ab. Although, the Dye-Ab labelled all of the intended target proteins, the Qdot-Ab was found bound to only some of the protein targets in the cytosol and could not reach the nucleus, even after extensive cell permeabilisation.

  5. 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.

  6. Compound specific radiocarbon dating for environmental pollutants

    Energy Technology Data Exchange (ETDEWEB)

    Schindler, Matthias; Kretschmer, Wolfgang; Scharf, Andreas; Kritzler, Karin; Stuhl, Alexander [Universitaet Erlangen-Nuernberg, Physikalisches Institut Abt. IV, Erwin-Rommel-Str.1, 91058 Erlangen (Germany)

    2013-07-01

    The talk gives a review about the procedures for measuring individual substances in complex matrices. Special focus will be on the usage of HPLC in separating those mixtures. There are explicit examples for the measurements of Polycyclic Aromatic Hydrocarbons and for carbonyls.

  7. Development of a Recombinant Cell-Based Indirect Immunofluorescence Assay for the Determination of Autoantibodies against Soluble Liver Antigen in Autoimmune Hepatitis

    Directory of Open Access Journals (Sweden)

    Christiane Radzimski

    2013-01-01

    Full Text Available Autoantibodies against soluble liver antigen (SLA are specific markers for autoimmune hepatitis (AIH type 1. In contrast to the determination of other AIH-associated autoantibodies by indirect immunofluorescence assay (IFA, detection of anti-SLA relied up to now on ELISA or immunoblot based on bacterially expressed recombinant protein. In order to develop a complementary IFA substrate, SLA isoform 1 was recombinantly produced in the human cell line HEK293 and controlled by a rabbit hyperimmune serum against SLA. The recombinant cells were used in IFA (RC-IFA to analyze sera from 20 AIH patients with anti-SLA positivity predetermined by ELISA together with 80 controls (20 anti-SLA negative AIH, 15 primary biliary cirrhosis, 15 HCV, and 30 healthy blood donors. Using RC-IFA, anti-SLA was detected in all ELISA positive AIH sera but in none of the controls. Furthermore, a cytosolic fraction of HEK293 containing SLA was able to neutralize the autoantibodies in all positive sera in a dose-dependent manner. HEK293 cells expressing SLA are a valid substrate for the serodiagnosis of AIH relevant autoantibodies by IFA. In concert with cryosections of primate liver, rat kidney, rat liver, rat stomach, and HEp-2 cells, they enable the parallel determination of all autoantibodies associated with autoimmune liver diseases.

  8. Compound Droplets on Fibers.

    Science.gov (United States)

    Weyer, Floriane; Ben Said, Marouen; Hötzer, Johannes; Berghoff, Marco; Dreesen, Laurent; Nestler, Britta; Vandewalle, Nicolas

    2015-07-21

    Droplets on fibers have been extensively studied in the recent years. Although the equilibrium shapes of simple droplets on fibers are well established, the situation becomes more complex for compound fluidic systems. Through experimental and numerical investigations, we show herein that compound droplets can be formed on fibers and that they adopt specific geometries. We focus on the various contact lines formed at the meeting of the different phases and we study their equilibrium state. It appears that, depending on the surface tensions, the triple contact lines can remain separate or merge together and form quadruple lines. The nature of the contact lines influences the behavior of the compound droplets on fibers. Indeed, both experimental and numerical results show that, during the detachment process, depending on whether the contact lines are triple or quadruple, the characteristic length is the inner droplet radius or the fiber radius.

  9. Detection of classic and invasive E. coli and Shigella serotypes in stools by indirect immunofluorescence

    Directory of Open Access Journals (Sweden)

    Waldely de Oliveira Dias

    1984-12-01

    Full Text Available IIF in the detection of invasive and classic enteropathogenic E. coli and Shigella serotypes was compared with traditional coproculture methods. IIP results agreed with the coproculture findings in 128 out of 140 cases tested for enteropathogenic E. coli (91% and in 108 out of 112 for Shigella (96%. All cases with positive reactions by coproculture were confirmed by IIP. In the control group it were obtained by IIF 12 cases with positive reactions for enteropathogenic E. coli and 4 cases for Shigella, including two cases of mixed infection by E. coli 026/Sh. dysenteriae and E. coli 0124/Sh. dysenteriae. It was discussed the high sensitivity and specificity of the IIF when compared with the traditional methods, being suggested that IIF is a valuable tool in epidemiological studies involving these organisms and an important aid in the stablishment of an early presumptive diagnosis of the acute infantile diarrhea.

  10. Magnesium compounds

    Science.gov (United States)

    Kramer, D.A.

    2007-01-01

    Seawater and natural brines accounted for about 52 percent of U.S. magnesium compounds production in 2006. Dead-burned magnesia was produced by Martin Marietta Magnesia Specialties from well brines in Michigan. Caustic-calcined magnesia was recovered from sea-water by Premier Chemicals in Florida; from well brines in Michigan by Martin Marietta and Rohm and Haas; and from magnesite in Nevada by Premier Chemicals. Intrepid Potash-Wendover and Great Salt Lake Minerals recovered magnesium chloride brines from the Great Salt Lake in Utah. Magnesium hydroxide was produced from brucite by Applied Chemical Magnesias in Texas, from seawater by SPI Pharma in Delaware and Premier Chemicals in Florida, and by Martin Marietta and Rohm and Haas from their operations mentioned above. About 59 percent of the magnesium compounds consumed in the United States was used for refractories that are used mainly to line steelmaking furnaces. The remaining 41 percent was consumed in agricultural, chemical, construction, environmental and industrial applications.

  11. Localization of H.pylori within the vacuole of Candida yeast by direct immunofluorescence technique.

    Science.gov (United States)

    Saniee, Parastoo; Siavoshi, Farideh; Nikbakht Broujeni, Gholamreza; Khormali, Mahmood; Sarrafnejad, Abdolfatah; Malekzadeh, Reza

    2013-12-01

    Reports indicate that H.pylori is able to invade the eukaryotic cells and establish inside their vacuoles. In this study, FITC-conjugated IgY-Hp was used to localize H.pylori inside the vacuole of Candida yeast. Presence of intracellular H.pylori inside the new generations of yeast cells was also examined by light microscopy and Live/Dead BacLight staining method. A single colony of fresh yeast culture was cultivated in a 100-µl medium containing yeast extract and N-acetylglucoseamine supplemented with fetal bovine serum. After 12-hr incubation at 37℃, FITC-conjugated IgY-Hp was added. After 3 hours, 10 µL of yeast suspension was smeared on a glass slide, air-dried and examined by fluorescent microscopy. Wet mounts of yeast culture and Live/Dead BacLight stained preparations were examined by light and fluorescent microscopy, respectively. Photographs were taken from the fast-moving H.pylori inside the yeast vacuoles. Fluorescent microscopy showed that FITC-conjugated IgY-Hp could enter yeast cells and specifically react with H.pylori, localizing the bacterium inside the yeast vacuole. Photographs taken from wet mounts observed by  light and fluorescent microscopy showed fast-moving H.pylori cells in the vacuole of mother as well as daughter yeast cells. The intravacuolar H.pylori cells stained green, showing their viability. Intracellular life of prokaryotes inside eukaryotes has been described as an evolutionary phenomenon with a great impact on bacterial persistence despite environmental stresses. Results of this study demonstrated the specific interaction of FITC-conjugated IgY-Hp with H.pylori cells and the bacterial localization inside the Candida yeast vacuole. The intracellular bacteria were viable and existed in the vacuole of next generations of yeast cells. It appears that H.pylori is well-equipped to dwell within the vacuole of eukaryotic cells where it is protected from stressful conditions, including antibacterial therapy. Presence of H

  12. pH-specific hydrothermal assembly of binary and ternary Pb(II)-(O,N-carboxylic acid) metal organic framework compounds: correlation of aqueous solution speciation with variable dimensionality solid-state lattice architecture and spectroscopic signatures.

    Science.gov (United States)

    Gabriel, C; Perikli, M; Raptopoulou, C P; Terzis, A; Psycharis, V; Mateescu, C; Jakusch, T; Kiss, T; Bertmer, M; Salifoglou, A

    2012-09-03

    Hydrothermal pH-specific reactivity in the binary/ternary systems of Pb(II) with the carboxylic acids N-hydroxyethyl-iminodiacetic acid (Heida), 1,3-diamino-2-hydroxypropane-N,N,N',N'-tetraacetic acid (Dpot), and 1,10-phenanthroline (Phen) afforded the new well-defined crystalline compounds [Pb(Heida)](n)·nH(2)O(1), [Pb(Phen)(Heida)]·4H(2)O(2), and [Pb(3)(NO(3))(Dpot)](n)(3). All compounds were characterized by elemental analysis, FT-IR, solution or/and solid-state NMR, and single-crystal X-ray diffraction. The structures in 1-2 reveal the presence of a Pb(II) center coordinated to one Heida ligand, with 1 exhibiting a two-dimensional (2D) lattice extending to a three-dimensional (3D) one through H-bonding interactions. The concurrent aqueous speciation study of the binary Pb(II)-Heida system projects species complementing the synthetic efforts, thereby lending credence to a global structural speciation strategy in investigating binary/ternary Pb(II)-Heida/Phen systems. The involvement of Phen in 2 projects the significance of nature and reactivity potential of N-aromatic chelators, disrupting the binary lattice in 1 and influencing the nature of the ultimately arising ternary 3D lattice. 3 is a ternary coordination polymer, where Pb(II)-Dpot coordination leads to a 2D metal-organic-framework material with unique architecture. The collective physicochemical properties of 1-3 formulate the salient features of variable dimensionality metal-organic-framework lattices in binary/ternary Pb(II)-(hydroxy-carboxylate) structures, based on which new Pb(II) materials with distinct architecture and spectroscopic signature can be rationally designed and pursued synthetically.

  13. Genetic, immunofluorescence labeling, and in situ hybridization techniques in identification of stem cells in male and female germline niches.

    Science.gov (United States)

    Singh, Shree Ram; Liu, Ying; Kango-Singh, Madhuri; Nevo, Eviatar

    2013-01-01

    Stem cells have an enormous capacity of self-renewal, as well as the ability to differentiate into specialized cell types. Proper control of these two properties of stem cells is crucial for animal development, growth control, and reproduction. Germline stem cells (GSCs) are a self-renewing population of germ cells, which generate haploid gametes (sperms or oocyte) that transmit genetic information from generation to generation. In Drosophila testis and ovary, GSCs are anchored around the niche cells. The cap cells cluster in females and hub cells in males act as a niche to control GSC behavior. With highly sophisticated genetic techniques in Drosophila, tremendous progress has been made in understanding the interactions between stem cells and niches at cellular and molecular levels. Here, we provide details of genetic, immunofluorescence labeling, and in situ hybridization techniques in identification and characterization of stem cells in Drosophila male and female germline niches.

  14. Application of indirect immunofluorescence to detection of Dientamoeba fragilis trophozoites in fecal specimens.

    Science.gov (United States)

    Chan, F T; Guan, M X; Mackenzie, A M

    1993-01-01

    An indirect fluorescent-antibody (IFA) assay was carried out to examine for the presence of Dientamoeba fragilis trophozoites in preserved fecal specimens. Antiserum to D. fragilis trophozoites was raised in a rabbit with a dixenic culture of D. fragilis (ATCC 30948) from the American Type Culture Collection. After absorption with Klebsiella pneumoniae and Bacteroides vulgatus, the immune rabbit serum was used for examination by the IFA assay. A total of 155 clinical samples were tested; 42 with no parasites, 9 with D. fragilis, and 104 with other parasites. The IFA assay identified seven D. fragilis organisms. Two specimens with doubtful IFA assay readings showed very scanty amounts of D. fragilis trophozoites on stained smears. There were no false-positive IFA assay readings. The IFA assay appeared to be a promising method because of its speed in screening. The specificity of the IFA assay indicates that other diagnostic tests such as an enzyme-linked immunosorbent assay could be developed to identify D. fragilis antigens in fecal specimens. Images PMID:8349746

  15. Furcation Perforation: Periradicular Tissue Response to Biodentine as a Repair Material by Histopathologic and Indirect Immunofluorescence Analyses.

    Science.gov (United States)

    Silva, Lea Assed Bezerra; Pieroni, Karina Alessandra Michelão Grecca; Nelson-Filho, Paulo; Silva, Raquel Assed Bezerra; Hernandéz-Gatón, Patrícia; Lucisano, Marília Pacífico; Paula-Silva, Francisco Wanderley Garcia; de Queiroz, Alexandra Mussolino

    2017-07-01

    The purpose of this study was to evaluate the in vivo response of periradicular tissues after sealing of furcation perforations with Biodentine, mineral trioxide aggregate (MTA), and gutta-percha by means of histopathologic and indirect immunofluorescence analyses. Thirty teeth of 3 dogs were divided into 3 groups: Biodentine (n = 14 teeth), MTA (negative control, n = 10 teeth), and gutta-percha (positive control, n = 6 teeth). After endodontic treatment, perforations were made on the center of the pulp chamber floor and filled with the materials. After 120 days, the animals were killed, and blocks containing the teeth and periradicular tissues were processed histotechnically for histopathologic semiquantitative (new mineralized tissue formation and bone resorption at the perforation site) and quantitative (thickness and area of newly formed mineralized tissue and number of inflammatory cells) analyses and RUNX2 immunofluorescence assay. Data were analyzed by χ 2 , Fisher exact test, Mann-Whitney test, one-way analysis of variance, Kruskal-Wallis test, and Dunn posttest (α = 0.05). MTA and Biodentine induced the formation of significantly more new mineralized tissue (P Biodentine and MTA exhibited no bone resorption in the furcation region, fewer inflammatory cells, and greater RUNX2 immunostaining intensity than gutta-percha. Although MTA presented higher frequency of complete sealing and greater thickness and area of newly formed mineralized tissue, Biodentine also had good histopathologic results and can be considered as an adequate furcation perforation repair material. Copyright © 2017 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  16. Magnesium compounds

    Science.gov (United States)

    Kramer, D.A.

    2012-01-01

    Seawater and natural brines accounted for about 57 percent of magnesium compounds produced in the United States in 2011. Dead-burned magnesia was produced by Martin Marietta Magnesia Specialties LLC from well brines in Michigan. Caustic-calcined magnesia was recovered from seawater by Premier Magnesia LLC in Florida, from well brines in Michigan by Martin Marietta and from magnesite in Nevada by Premier Magnesia. Intrepid Potash Wendover LLC and Great Salt Lake Minerals Corp. recovered magnesium chloride brines from the Great Salt Lake in Utah. Magnesium hydroxide was produced from seawater by SPI Pharma Inc. in Delaware and Premier Magnesia in Florida, and by Martin Marietta from its brine operation in Michigan.

  17. Bismaleimide compounds

    Science.gov (United States)

    Adams, Johnnie E.; Jamieson, Donald R.

    1986-01-14

    Bismaleimides of the formula ##STR1## wherein R.sub.1 and R.sub.2 each independently is H, C.sub.1-4 -alkyl, C.sub.1-4 -alkoxy, C1 or Br, or R.sub.1 and R.sub.2 together form a fused 6-membered hydrocarbon aromatic ring, with the proviso that R.sub.1 and R.sub.2 are not t-butyl or t-butoxy; X is O, S or Se; n is 1-3; and the alkylene bridging group, optionally, is substituted by 1-3 methyl groups or by fluorine, form polybismaleimide resins which have valuable physical properties. Uniquely, these compounds permit extended cure times, i.e., they remain fluid for a time sufficient to permit the formation of a homogeneous melt prior to curing.

  18. The nature of compounds: a psychocentric perspective.

    Science.gov (United States)

    Libben, Gary

    2014-01-01

    Although compound words often seem to be words that themselves contain words, this paper argues that this is not the case for the vast majority of lexicalized compounds. Rather, it is claimed that as a result of acts of lexical processing, the constituents of compound words develop into new lexical representations. These representations are bound to specific morphological roles and positions (e.g., head, modifier) within a compound word. The development of these positionally bound compound constituents creates a rich network of lexical knowledge that facilitates compound processing and also creates some of the well-documented patterns in the psycholinguistic and neurolinguistic study of compounding.

  19. atmospheric volatile organic compounds

    Directory of Open Access Journals (Sweden)

    A. R. Koss

    2016-07-01

    organic compounds (VOCs that cannot be ionized with H3O+ ions (e.g., in a PTR-MS or H3O+ CIMS instrument. Here we describe the adaptation of a high-resolution time-of-flight H3O+ CIMS instrument to use NO+ primary ion chemistry. We evaluate the NO+ technique with respect to compound specificity, sensitivity, and VOC species measured compared to H3O+. The evaluation is established by a series of experiments including laboratory investigation using a gas-chromatography (GC interface, in situ measurement of urban air using a GC interface, and direct in situ measurement of urban air. The main findings are that (1 NO+ is useful for isomerically resolved measurements of carbonyl species; (2 NO+ can achieve sensitive detection of small (C4–C8 branched alkanes but is not unambiguous for most; and (3 compound-specific measurement of some alkanes, especially isopentane, methylpentane, and high-mass (C12–C15 n-alkanes, is possible with NO+. We also demonstrate fast in situ chemically specific measurements of C12 to C15 alkanes in ambient air.

  20. An estimation of Central Iberian Peninsula atmospheric δ13C and water δD in the Upper Cretaceous using pyrolysis compound specific isotopic analysis (Py-CSIA) of a fossil conifer.

    Science.gov (United States)

    González-Pérez, José A.; Jiménez-Morillo, Nicasio T.; De la Rosa, José M.; Almendros, Gonzalo; González-Vila, Francisco J.

    2015-04-01

    Frenelopsis is a frequently found genus of the Cretaceous floras adapted to dry, saline and in general to environmental conditions marked by severe water stress [1]. Stable isotope analysis of fossil organic materials can be used to infer palaeoenvironmental variables helpful to reconstruct plant paleohabitats [2]. In this study stable isotope analysis of organic fossil remains (FR) and humic fractions (FA, HA and humin) of Frenelopsis oligiostomata are studied in bulk (C, H, O, N IRMS) and in specific compounds released by pyrolysis (C, H, Py-CSIA). Well preserved F. oligiostomata fossils were handpicked from a limestone included in compacted marls from Upper Cretaceous (Senonian c. 72 Mya) in Guadalix de la Sierra (Madrid, Spain) [3]. The fossils were decarbonated with 6M HCl. Humic substances were extracted from finely ground fossil remains (FR) by successive treatments with 0.1M Na4P2O7 + NaOH [4]. The extract was acidified resulting into insoluble HA and soluble FA fractions. The HA and FA were purified as in [5] and [6] respectively. Bulk stable isotopic analysis (δ13C, δD, δ18O, δ15N IRMS) was done in an elemental micro-analyser coupled to a continuous flow Delta V Advantage isotope ratio mass spectrometer (IRMS). Pyrolysis compound specific isotopic analysis Py-CSIA (δ13C, δD): was done by coupling a double-shot pyrolyzer to a chromatograph connected to an IRMS. Structural features of specific peaks were inferred by comparing/matching mass spectra from conventional Py-GC/MS (data not shown) with Py-GC/IRMS chromatograms obtained using the same chromatographic conditions. Bulk C isotopic signature found for FR (-20.5±0.02 ‰) was in accordance with previous studies [2, 7-9]. This heavy isotopic δ13C signature indicates a depleted stomatal conductance and paleoenvironmental growth conditions of water and salt stress. This is in line with the morphological and depositional characteristics [3] confirming that F. oligostomata was adapted to highly xeric

  1. Development and validation of an universal interface for compound-specific stable isotope analysis of chlorine (37Cl/35Cl) by GC-high-temperature conversion (HTC)-MS/IRMS.

    Science.gov (United States)

    Renpenning, Julian; Hitzfeld, Kristina L; Gilevska, Tetyana; Nijenhuis, Ivonne; Gehre, Matthias; Richnow, Hans-Hermann

    2015-03-03

    A universal application of compound-specific isotope analysis of chlorine was thus far limited by the availability of suitable analysis techniques. In this study, gas chromatography in combination with a high-temperature conversion interface (GC-HTC), converting organic chlorine in the presence of H2 to gaseous HCl, was coupled to a dual-detection system, combining an ion trap mass spectrometer (MS) and isotope-ratio mass spectrometer (IRMS). The combination of the MS/IRMS detection enabled a detailed characterization, optimization, and online monitoring of the high-temperature conversion process via ion trap MS as well as a simultaneous chlorine isotope analysis by the IRMS. Using GC-HTC-MS/IRMS, chlorine isotope analysis at optimized conversion conditions resulted in very accurate isotope values (δ(37)Cl(SMOC)) for measured reference material with known isotope composition, including chlorinated ethylene, chloromethane, hexachlorocyclohexane, and trichloroacetic acids methyl ester. Respective detection limits were determined to be <15 nmol Cl on column with achieved precision of <0.3‰.

  2. Immunofluorescence assay in India for confirmation of HIV-1 infection using a T-cell line infected with defective HIV-1.

    Science.gov (United States)

    Bala, Manju; Arias, Juan F; Deb, Monorama; Ikuta, Kazuyoshi

    2010-12-01

    In India, the enzyme immunoassay (EIA)/rapid test is used for screening and confirmatory antibody testing of HIV infection, and all HIV reactive samples are further confirmed by two other rapid tests working on different principles; however, Western blotting (WB) and immunofluorescence (IF) assays are not routinely performed in this country. A total of 2104 sera from Indian subjects were tested for the presence of HIV-1 antibody using EIA/rapid tests, according to the guidelines of the National AIDS Control Organization of India, and were also subjected to IF test using L-2 cells persistently infected with defective HIV-1. WB and a nested reverse transcriptase polymerase chain reaction (RT-PCR) were performed on discrepant samples. IF results were 100% concordant with EIA/rapid tests for 212 HIV-1-positive samples and 1889 HIV-1-negative samples. Interestingly, three (0.14%) samples negative by EIA/rapid tests were weakly or moderately positive (1+/2+) by IF test. All three of these samples were confirmed to be negative by WB (reactive with Gag/Pol, but not with Env), but positive by RT-PCR with primers targeting the C2-V5 fragment of the env gene. These three samples were from individuals who voluntarily reported for HIV testing because of high-risk practices, and they may have been at an early stage of HIV infection. These results confirm that the IF test using L-2 cells is a sensitive and specific alternative method for confirmation of HIV-1 infection and could be included in the diagnostic algorithm in reference laboratories in developing countries. Copyright © 2010 International Society for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

  3. Evaluation of Immunofluorescence Antibody Test Used for the Diagnosis of Canine Leishmaniasis in the Mediterranean Basin: A Systematic Review and Meta-Analysis.

    Directory of Open Access Journals (Sweden)

    Amel Adel

    Full Text Available With an expected sensitivity (Se of 96% and specificity (Sp of 98%, the immunofluorescence antibody test (IFAT is frequently used as a reference test to validate new diagnostic methods and estimate the canine leihmaniasis (CanL true prevalence in the Mediterranean basin. To review the diagnostic accuracy of IFAT to diagnose CanL in this area with reference to its Se and Sp and elucidate the potential causes of their variations, a systematic review was conducted (31 studies for the 26-year period. Three IFAT validation methods stood out: the classical contingency table method, methods based on statistical models and those based on experimental studies. A variation in the IFAT Se and Sp values and cut-off values was observed. For the classical validation method based on a meta-analysis, the Se of IFAT was estimated in this area as 89.86% and 31.25% in symptomatic and asymptomatic dogs, respectively. The Sp of IFAT was estimated in non-endemic and endemic areas as 98.12% and 96.57%, respectively. IFAT can be considered as a good standard test in non-endemic areas for CanL, but its accuracy declines in endemic areas due to the complexity of the disease. Indeed, the accuracy of IFAT is due to the negative results obtained in non-infected dogs from non-endemic areas and to the positive results obtained in sera of symptomatic dogs living in endemic areas. But IFAT results are not unequivocal when it comes to determining CanL infection on asymptomatic dogs living in endemic areas. Statistical methods might be a solution to overcome the lack of gold standard, to better categorize groups of animals investigated, to assess optimal cut-off values and to allow a better estimate of the true prevalence aiming information on preventive/control measures for CanL.

  4. IMMUNOFLUORESCENCE IN CLINICAL BIOLOGY,

    Science.gov (United States)

    SERODIAGNOSIS), (*FLUORESCENT ANTIBODY TECHNIQUES , IDENTIFICATION, BACTERIA, VIRUSES, TREPONEMA PALLIDUM, NUCLEI( BIOLOGY ), ANTIGEN ANTIBODY REACTIONS, BLOOD GROUPS, ERYTHROCYTES, TOXOPLASMA, IMMUNOLOGY, BELGIUM.

  5. Hausa verbal compounds

    NARCIS (Netherlands)

    McIntyre, Joseph Anthony

    2006-01-01

    Verbal compounds abound in Hausa (a Chadic language). A very broad definition of Hausa verbal compounds (henceforth: VC) is “a compound with a verb”. Four types of verbal compound are analysed: V[erb]+X compounds, PAC+V compounds (a PAC is a pronoun complex indicating TAM), VCs with a ma prefix

  6. Demineralization of dentin with EDTA in organic solvent: immunofluorescence of collagen in osteogenesis imperfecta and normal teeth.

    Science.gov (United States)

    Lukinmaa, P L; Ranta, H; Ranta, K; Peltonen, L; Hietanen, J

    1985-12-01

    The immunofluorescence of dentin collagen(s) after demineralization with ethanolic trimethylammonium ethylenediaminetetra-acetic acid (EtOH-TMA-EDTA) was studied using normal deciduous teeth, and deciduous and permanent teeth from three patients with Osteogenesis imperfecta (OI). Sections of the demineralized teeth were immunostained with anti type I, III and IV collagen sera. Preservation of the antigenicity of the collagens after treatment with EtOH-TMA-EDTA was confirmed by staining of the soft tissue controls. Anti type I collagen sera stained the normal dentin matrix peritubularly only. In OI, a homogeneous fluorescence of the mantle dentin and the dentin zone surrounding the abnormal canal-like structures was observed. With anti type III collagen serum, the normal dentin matrix failed to stain. In OI, the staining pattern was a narrow halo surrounding the canal-like structures. Alteration in the collagen or the noncollagenous components of the dentin matrix may explain the staining reactions of the various collagens in OI.

  7. RRM1, ERCC1 and TS1 Immunofluorescence Expression in Leiomyosarcoma: A Tissue Microarray Study with Clinical Outcome Correlation Analysis.

    Science.gov (United States)

    Zheng, Sam D; Bui, Katherine; Chiappori, Alberto; Bepler, Gerold; Bui, Marilyn M

    2016-07-01

    ERCC1, RRM1 and TS1 are reportedly linked to chemotherapy resistance in lung and other cancers. However, there are currently no studies reporting the relationship between these genes and clinical parameters in leiomyosarcomas. This study investigated the expression pattern of ERCC1, RRM1 and TS1 in forty-four leiomyosarcoma samples by the use of tissue microarray (TMA), immunofluorescence and AQUA methods. The results were then analyzed for expression level and correlations were made with clinical outcome to determine their potential prognostic value in leiomyosarcoma. In the forty-four samples studied, the expression level of these three proteins can be well quantified in the AQUA system and reflected by the AQUA score. RRM1 and ERCC1 expression levels did not show any relationship with overall survival. However, a correlation was found between TS1 expression in the cytoplasm and overall survival. The high expression group had a shorter overall survival time (log-rank p = 0.0498). This trend was confirmed by the Cox proportional hazards model. The poor overall survival of leiomyosarcoma is linked to TS1 cytoplasm expression which may be useful in predicting prognoses of this tumor, methods targeting expression of TS1 may lead to improved overall survival in leiomyosarcoma, though more detailed information regarding treatment information and a larger sample size is needed to confirm this phenomenon.

  8. Detection of antibodies to Plasmodium vivax by indirect immunofluorescence: influence of the geographic origin of antigens and serum samples.

    Science.gov (United States)

    Benzerroug, E H; Demedts, P; Wéry, M

    1986-03-01

    The results of a double-blind serological study of 15 sera sampled in a residual focus of vivax malaria transmission in Algeria, and of 7 sera from patients with slide-proven P. vivax infections acquired in India, are analyzed. The reactivity of each of these serum samples was tested by indirect immunofluorescence using 6 different batches of antigen, including 3 batches of P. vivax antigen prepared with isolates from Zaire (Africa), India and the Solomon Islands, respectively. The geometric mean of reciprocal titers (GMRT) calculated on the 7 sera from proven vivax infections fell from 289.8 using the homologous antigen from the same geographic origin (India) to 48.7 using a homologous (vivax) antigen originating from a different continent (Africa). Among the 15 samples from Algeria, the percentage of seropositives decreased from 100% using the homologous P. vivax antigen originating from the same continent (Africa) to 53.3% using a homologous antigen from India. Two aspects are included in the discussion: in seroepidemiological studies, sensitivity could be improved by the use of a homologous antigen from the same geographic origin; in detection of clinical cases of malaria and species identification based on serology, our results stress the need for caution in interpreting serological titers and for taking into account the geographic origin of the isolates used as antigen.

  9. 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.

  10. An immunofluorescence assay for extracellular matrix components highlights the role of epithelial cells in producing a stable, fibrillar extracellular matrix.

    Science.gov (United States)

    Qureshi, Omar S; Bon, Hélène; Twomey, Breda; Holdsworth, Gill; Ford, Kirsty; Bergin, Marianne; Huang, Linghong; Muzylak, Mariusz; Healy, Louise J; Hurdowar, Vanessa; Johnson, Timothy S

    2017-10-15

    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. © 2017. Published by The Company of Biologists Ltd.

  11. Magnetochemistry: Compounds and Concepts

    Science.gov (United States)

    Lueken, Heiko; Schilder, Helmut; Eifert, Thomas; Handrick, Klaus; Huening, Felix

    New materials in today's magnetochemistry are reviewed, among them spin-crossover compounds, single-molecule magnets, supramolecular magnetic materials, ferrofluids, and ferromagnetic nanocrystal superlattices, each with its specific application potential. Advanced computer programs are available which consider all relevant perturbations on the 'magnetic electrons' and allow, on the basis of precise measurement results, the evaluation of magnetic parameters, above all the exchange parameter J. By means of molecular, ionic, and metallic compounds the J values are discussed with respect to sign and magnitude on the basis of concepts which take into consideration overlap of magnetic orbitals (direct exchange, superexchange) and the conduction electron concentration. Finally, further developments in the field of magnetochemistry are prospected.

  12. 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.

  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. RNA-Seq analysis and annotation of a draft blueberry genome assembly identifies candidate genes involved in fruit ripening, biosynthesis of bioactive compounds, and stage-specific alternative splicing

    National Research Council Canada - National Science Library

    Gupta, Vikas; Estrada, April D; Blakley, Ivory; Reid, Rob; Patel, Ketan; Meyer, Mason D; Andersen, Stig Uggerhøj; Brown, Allan F; Lila, Mary Ann; Loraine, Ann E

    2015-01-01

    ... from other species, typically the grape Vitis vinifera. Comparison of gene models to the PlantCyc database of metabolic pathway enzymes identified candidate genes involved in synthesis of bioactive compounds, including...

  15. Multiplex RT-PCR and indirect immunofluorescence assays for detection and subtyping of human influenza virus in Tunisia.

    Science.gov (United States)

    Ben M'hadheb, Manel; Harrabi, Myriam; Souii, Amira; Jrad-Battikh, Nadia; Gharbi, Jawhar

    2015-03-01

    Influenza viruses are negative stranded segmented RNA viruses belonging to Orthomyxoviridae family. They are classified into three types A, B, and C. Type A influenza viruses are classified into subtypes according to the antigenic characters of the surface glycoproteins: hemagglutinin (H) and neuraminidase (N). The aim of the present study is to develop a fast and reliable multiplex RT-PCR technique for detecting simultaneously the subtypes A/H1N1 and A/H3N2 of influenza virus. Our study included 398 patients (mean age 30.33 ± 19.92 years) with flu or flu-like syndromes, consulting physicians affiliated with collaborating teams. A multiplex RT-PCR detecting A/H1N1 and A/H3N2 influenza viruses and an examination by indirect immunofluorescence (IFI) were performed. In the optimized conditions, we diagnosed by IFI a viral infection in 90 patients (22.6 %): 85 cases of influenza type A, four cases of influenza type B, and only one case of coinfection with types A and B. An evaluation of the technique was performed on 19 clinical specimens positive in IFI, and we detected eight cases of A/H3N2, five cases of A/H1N1, one case of influenza virus type A which is not an H1N1 nor H3N2, and five negative cases. Multiplex RT-PCR is a sensitive technique allowing an effective and fast diagnosis of respiratory infections caused by influenza viruses in which the optimization often collides with problems of sensibility.

  16. Composite biomarkers defined by multiparametric immunofluorescence analysis identify ALK-positive adenocarcinoma as a potential target for immunotherapy.

    Science.gov (United States)

    Roussel, Hélène; De Guillebon, Eléonore; Biard, Lucie; Mandavit, Marion; Gibault, Laure; Fabre, Elisabeth; Antoine, Martine; Hofman, Paul; Beau-Faller, Michèle; Blons, Hélène; Danel, Claire; Barthes, Françoise Le Pimpec; Gey, Alain; Granier, Clémence; Wislez, Marie; Laurent-Puig, Pierre; Oudard, Stéphane; Bruneval, Patrick; Badoual, Cécile; Cadranel, Jacques; Tartour, Eric

    2017-01-01

    Anaplastic lymphoma kinase (ALK) inhibitors have been successfully developed for non-small cell lung carcinoma (NSCLC) displaying chromosomal rearrangements of the ALK gene, but unfortunately resistance invariably occurs. Blockade of the PD-1-PD-L1/2 inhibitory pathway constitutes a breakthrough for the treatment of NSCLC. Some predictive biomarkers of clinical response to this therapy are starting to emerge, such as PD-L1 expression by tumor/stromal cells and infiltration by CD8+ T cells expressing PD-1. To more effectively integrate all of these potential biomarkers of clinical response to immunotherapy, we have developed a multiparametric immunofluorescence technique with automated immune cell counting to comprehensively analyze the tumor microenvironment of ALK-positive adenocarcinoma (ADC). When analyzed as either a continuous or a dichotomous variable, the mean number of tumor cells expressing PD-L1 (p = 0.012) and the percentage of tumor cells expressing PD-L1 were higher in ALK-positive ADC than in EGFR-mutated ADC or WT (non-EGFR-mutated and non-KRAS-mutated) NSCLC. A very strong correlation between PD-L1 expression on tumor cells and intratumoral infiltration by CD8+ T cells was observed, suggesting that an adaptive mechanism may partly regulate this expression. A higher frequency of tumors combining positive PD-L1 expression and infiltration by intratumoral CD8+ T cells or PD-1+CD8+ T cells was also observed in ALK-positive lung cancer patients compared with EGFR-mutated (p = 0.03) or WT patients (p = 0.012). These results strongly suggest that a subgroup of ALK-positive lung cancer patients may constitute good candidates for anti-PD-1/-PD-L1 therapies.

  17. Quantitative immunofluorescence microscopy of subcellular GLUT4 distribution in human skeletal muscle: effects of endurance and sprint interval training

    Science.gov (United States)

    Bradley, Helen; Shaw, Christopher S.; Worthington, Philip L.; Shepherd, Sam O.; Cocks, Matthew; Wagenmakers, Anton J. M.

    2014-01-01

    Abstract Increases in insulin‐mediated glucose uptake following endurance training (ET) and sprint interval training (SIT) have in part been attributed to concomitant increases in glucose transporter 4 (GLUT4) protein content in skeletal muscle. This study used an immunofluorescence microscopy method to investigate changes in subcellular GLUT4 distribution and content following ET and SIT. Percutaneous muscle biopsy samples were taken from the m. vastus lateralis of 16 sedentary males in the overnight fasted state before and after 6 weeks of ET and SIT. An antibody was fully validated and used to show large (> 1 μm) and smaller (<1 μm) GLUT4‐containing clusters. The large clusters likely represent trans‐Golgi network stores and the smaller clusters endosomal stores and GLUT4 storage vesicles (GSVs). Density of GLUT4 clusters was higher at the fibre periphery especially in perinuclear regions. A less dense punctate distribution was seen in the rest of the muscle fibre. Total GLUT4 fluorescence intensity increased in type I and type II fibres following both ET and SIT. Large GLUT4 clusters increased in number and size in both type I and type II fibres, while the smaller clusters increased in size. The greatest increases in GLUT4 fluorescence intensity occurred within the 1 μm layer immediately adjacent to the PM. The increase in peripheral localisation and protein content of GLUT4 following ET and SIT is likely to contribute to the improvements in glucose homeostasis observed after both training modes. PMID:25052490

  18. Relationship of Serum Levels of Anti-Desmoglein Antibodies and Direct Immunofluorescence Findings with Clinical Activity of Pemphigus

    Directory of Open Access Journals (Sweden)

    Mediha Yılmaz

    Full Text Available Background and Design: Pemphigus is an autoimmune disease that results in blistering of the skin and mucous membranes. In this study, we investigated the relationship between disease activity and remission with ELISA scores and direct immunofluorescence (IF - two methods used for the detection of antibodies against desmoglein-1 (dsg-1 and desmoglein-3 (dsg-3 that are responsible for blister formation. Material and Method: Twenty-three pemphigus vulgaris patients and two pemphigus foliaceus patients were enrolled in the study. The serum levels of anti-dsg-1 and anti-dsg-3 antibodies were measured with ELISA before therapy and at 3, 6, and 12 month of clinical remission. Concurrently, direct IF was performed on perilesional skin during active disease and on normal buttock skin/lower lip mucosa in remission. The tests were repeated if relapse has occured. Results: Anti-dsg-1 was detected in 17 (73.9% pemphigus vulgaris patients and anti-dsg-3 in 23 (100% pemphigus vulgaris patients. In two pemphigus foliaceus patients, anti-dsg-1 values were positive, while anti-dsg-3 values were negative. A statistically significant correlation was seen between anti-dsg-1 antibody serum levels and skin severity scores (r: 0.577; p: 0.003, as well as between anti-dsg-3 antibody serum levels and oral mucosa severity scores (r: 0.539; p: 0.008. Direct IF results in 16 patients (84.2% who achieved complete remission were negative. In 9 patients who relapsed, elevated serum values of anti-dsg-1 and/or anti-dsg-3 were also found. Increase in serum antibody levels was detected 1-4 months before the relapse in three of them. Conclusion: In this study, we observed that serum desmoglein antibody levels correlated with disease severity and activity. In clinical remission, serial measurements of desmoglein antibodies can provide a guide for clinical follow-up and treatment modification.

  19. Rubber compounding and processing

    CSIR Research Space (South Africa)

    John, MJ

    2014-06-01

    Full Text Available This chapter presents an overview on the compounding and processing techniques of natural rubber compounds. The introductory portion deals with different types of rubbers and principles of rubber compounding. The primary and secondary fillers used...

  20. Comparison of Genital Chlamydia trachomatis Infection Incidence Between Women With Infertility and Healthy Women in Iran Using PCR and Immunofluorescence Methods.

    Science.gov (United States)

    Marashi, Seyed Mahmoud Amin; Moulana, Zahra; Imani Fooladi, Abbas Ali; Mashhadi Karim, Mohammad

    2014-04-01

    For a long time, infertility has been one of the most sequels in medical sciences with microbial agents as one group of its causes. The possible etiological role of Chlamydia trachomatis in infertility was suggested years ago, but it has not yet been proved completely. To decrease the severe involvements of C. trachomatis infections, screening by efficient diagnostic methods are necessary. In this study we attempted to determine the incidence of C. trachomatis in infertile women and compared this with healthy women. This case-control study was performed on 150 infertile women with unknown causes and without physiological deficiency for infertility. The control group consisted of 200 fertile safe and impregnated women. Presence of C. trachomatis in the two groups was examined by direct and indirect immunofluorescence tests and PCR. C. trachomatis was detected by direct immunofluorescence method in 23 (15.3%) infertile women compared and 7 (3.5%) healthy controls. Using indirect immunofluorescence tests, a positive test titer of 1:16 as well as the above results were detected in 34 (22.6%) of the infertile cases and 9 (4.5%) of the controls. C. trachomatis was detected by PCR method in 48 (32%) infertile women and 13 (8.7%) among the controls. The results of our study suggest that there is a significant association between C. trachomatis infection and female infertility.

  1. Antigen-based immunofluorescence analysis of B-cell targeting: advanced technology for the generation of novel monoclonal antibodies with high efficiency and selectivity.

    Science.gov (United States)

    Tomita, Masahiro; Fukuda, Takatomi; Ozu, Akira; Kimura, Kei-ichi; Tsong, Tian Yow; Yoshimura, Tetsuro

    2006-10-01

    On the basis of immunofluorescence analysis, an entire pathway of B-cell targeting was successfully identified, which can drive selective production of monoclonal antibodies with high efficiency and selectivity. The technique comprises three critical steps, antigen-based preselection of B lymphocytes, formation of antigenselected B lymphocyte and myeloma cell complexes, and selective fusion of B-cell-myeloma cell complexes with electrical pulses. Intriguingly, expression of surface immunoglobulin receptors on B lymphocytes was recognized even after immunization in vitro. The number of the antigen-selected B lymphocytes after in vitro immunization was in fact higher than that obtained after in vivo immunization, suggesting that such shortterm immunization is applicable for B-cell targeting. Immunofluorescence analysis revealed the targeting technique to demonstrate fivefold to tenfold higher efficiency for formation of hybridoma cells than a polyethylene glycol (PEG)-mediated method. This efficiency is in good agreement with production of hybridoma cells secreting desired monoclonal antibodies determined by an enzyme-linked immunosorbent assay (ELISA). The addition of a low concentration of PEG brought about enhanced fusion efficiency and reduced cell damage at even electric intensities as high as 4.0 kV/cm and 5.0 kV/cm. Here we demonstrate that immunofluorescence analysis can successfully clarify an entire pathway of B-cell targeting and provide advantages over the PEG-mediated method. This advanced technology may be applicable for rapid production of monoclonal antibodies based on in vitro immunization.

  2. Patterns of glomerular disease based on 4-year kidney biopsy material analyzed by light microscopy and immunofluorescence: a retrospective single-center analysis in Poland

    Directory of Open Access Journals (Sweden)

    Anna Olewicz-Gawlik

    2017-02-01

    Full Text Available The aim of this study was to assess the epidemiology of different patterns of chronic glomerular diseases based on clinical, histopathological and immunofluorescent findings of glomerulonephritis patients hospitalized in the Department of Nephrology, Transplantology and Internal Diseases in Poznan between January 2009 and December 2012. We retrospectively studied 418 patients who had been subjected to renal biopsies. Data on serum creatinine concentration, 24 h proteinuria, arterial hypertension, diabetes mellitus, and histological and immunofluorescent findings were collected. The patients’ mean age was 42 ±15. The male sex prevailed (53.1%. Immunoglobulin A nephropathy was the most common finding (18.9%, followed by focal segmental glomerulosclerosis (16.3%, membranous glomerulonephritis (10.1%, lupus nephritis (8.4%, extracapillary glomerulonephritis (3.3% and membranoproliferative glomerulonephritis (2.6%. In 69 (16.5% patients the biopsy was non-informative or non-diagnostic. Patients with membranous nephropathy presented the highest frequency of nephrotic syndrome (71.4%, followed by membranoproliferative glomerulonephritis and focal segmental glomerulosclerosis. Combined analysis of the clinical, histopathological and immunofluorescent findings in glomerulonephritis patients based on a single center’s data can provide important epidemiological findings.

  3. Compound Cuing in Free Recall

    Science.gov (United States)

    Lohnas, Lynn J.; Kahana, Michael J.

    2014-01-01

    According to the retrieved context theory of episodic memory, the cue for recall of an item is a weighted sum of recently activated cognitive states, including previously recalled and studied items as well as their associations. We show that this theory predicts there should be compound cuing in free recall. Specifically, the temporal contiguity…

  4. Comparison of a commercial qualitative real-time RT-PCR kit with direct immunofluorescence assay (DFA) and cell culture for detection of influenza A and B in children.

    Science.gov (United States)

    Gharabaghi, Farhad; Tellier, Raymond; Cheung, Rose; Collins, Carol; Broukhanski, George; Drews, Steven J; Richardson, Susan E

    2008-06-01

    Institutional pandemic planning prompted a study of the molecular detection of influenza virus from respiratory specimens in children, compared to conventional diagnostics. To evaluate the performance of a commercial qualitative real-time RT-PCR kit (rRT-PCR), the artus Influenza LC RT-PCR (Qiagen). STUDY DESIGN (METHODS): Specimens were pre-selected to include a high percentage of positives by direct immunofluorescence assay (DFA) or culture. The sensitivity and specificity of the kit for detection of influenza A and B in children were determined against the gold standard, DFA and culture. Specimens yielding discordant results between artus and the gold standard were tested against a reference rRT-PCR assay (Centers for Disease Control) to create an "expanded gold standard". When compared to DFA or cell culture, the sensitivity of the rRT-PCR artus kit was 96.2% and the specificity was 94%. It detected influenza RNA in 6.0% of clinical samples negative by DFA or culture. Using the expanded gold standard, the revised sensitivity was 98.7% (98.6% for influenza A and 97.6% for influenza B) and the specificity was 100%. The artus Influenza LC RT-PCR kit is an effective alternative to virus isolation and DFA for the detection of influenza A and B in pediatric clinical specimens.

  5. Antibacterial and Antifungal Compounds from Marine Fungi

    OpenAIRE

    Xu, Lijian; Meng, Wei; Cao, Cong; Wang, Jian; Shan, Wenjun; Wang, Qinggui

    2015-01-01

    This paper reviews 116 new compounds with antifungal or antibacterial activities as well as 169 other known antimicrobial compounds, with a specific focus on January 2010 through March 2015. Furthermore, the phylogeny of the fungi producing these antibacterial or antifungal compounds was analyzed. The new methods used to isolate marine fungi that possess antibacterial or antifungal activities as well as the relationship between structure and activity are shown in this review.

  6. Antibacterial and Antifungal Compounds from Marine Fungi

    Science.gov (United States)

    Xu, Lijian; Meng, Wei; Cao, Cong; Wang, Jian; Shan, Wenjun; Wang, Qinggui

    2015-01-01

    This paper reviews 116 new compounds with antifungal or antibacterial activities as well as 169 other known antimicrobial compounds, with a specific focus on January 2010 through March 2015. Furthermore, the phylogeny of the fungi producing these antibacterial or antifungal compounds was analyzed. The new methods used to isolate marine fungi that possess antibacterial or antifungal activities as well as the relationship between structure and activity are shown in this review. PMID:26042616

  7. RNA-Seq analysis and annotation of a draft blueberry genome assembly identifies candidate genes involved in fruit ripening, biosynthesis of bioactive compounds, and stage-specific alternative splicing.

    Science.gov (United States)

    Gupta, Vikas; Estrada, April D; Blakley, Ivory; Reid, Rob; Patel, Ketan; Meyer, Mason D; Andersen, Stig Uggerhøj; Brown, Allan F; Lila, Mary Ann; Loraine, Ann E

    2015-01-01

    Blueberries are a rich source of antioxidants and other beneficial compounds that can protect against disease. Identifying genes involved in synthesis of bioactive compounds could enable the breeding of berry varieties with enhanced health benefits. Toward this end, we annotated a previously sequenced draft blueberry genome assembly using RNA-Seq data from five stages of berry fruit development and ripening. Genome-guided assembly of RNA-Seq read alignments combined with output from ab initio gene finders produced around 60,000 gene models, of which more than half were similar to proteins from other species, typically the grape Vitis vinifera. Comparison of gene models to the PlantCyc database of metabolic pathway enzymes identified candidate genes involved in synthesis of bioactive compounds, including bixin, an apocarotenoid with potential disease-fighting properties, and defense-related cyanogenic glycosides, which are toxic. Cyanogenic glycoside (CG) biosynthetic enzymes were highly expressed in green fruit, and a candidate CG detoxification enzyme was up-regulated during fruit ripening. Candidate genes for ethylene, anthocyanin, and 400 other biosynthetic pathways were also identified. Homology-based annotation using Blast2GO and InterPro assigned Gene Ontology terms to around 15,000 genes. RNA-Seq expression profiling showed that blueberry growth, maturation, and ripening involve dynamic gene expression changes, including coordinated up- and down-regulation of metabolic pathway enzymes and transcriptional regulators. Analysis of RNA-seq alignments identified developmentally regulated alternative splicing, promoter use, and 3' end formation. We report genome sequence, gene models, functional annotations, and RNA-Seq expression data that provide an important new resource enabling high throughput studies in blueberry.

  8. Promiscuity progression of bioactive compounds over time.

    Science.gov (United States)

    Hu, Ye; Jasial, Swarit; Bajorath, Jürgen

    2015-01-01

    In the context of polypharmacology, compound promiscuity is rationalized as the ability of small molecules to specifically interact with multiple targets. To study promiscuity progression of bioactive compounds in detail, nearly 1 million compounds and more than 5.2 million activity records were analyzed. Compound sets were assembled by applying different data confidence criteria and selecting compounds with activity histories over many years. On the basis of release dates, compounds and activity records were organized on a time course, which ultimately enabled monitoring data growth and promiscuity progression over nearly 40 years, beginning in 1976. Surprisingly low degrees of promiscuity were consistently detected for all compound sets and there were only small increases in promiscuity over time. In fact, most compounds had a constant degree of promiscuity, including compounds with an activity history of 10 or 20 years. Moreover, during periods of massive data growth, beginning in 2007, promiscuity degrees also remained constant or displayed only minor increases, depending on the activity data confidence levels. Considering high-confidence data, bioactive compounds currently interact with 1.5 targets on average, regardless of their origins, and display essentially constant degrees of promiscuity over time. Taken together, our findings provide expectation values for promiscuity progression and magnitudes among bioactive compounds as activity data further grow.

  9. Bioactive compounds in berries relevant to human health

    NARCIS (Netherlands)

    Battino, M.; Beekwilder, M.J.; Denoyes-Rothan, B.; Laimer, M.

    2009-01-01

    Berries contain powerful antioxidants, potential allergens, and other bioactive compounds. Genetic and environmental factors affect production and storage of such compounds. For this reason breeding and biotechnological approaches are currently used to control or to increase the content of specific

  10. 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.

  11. FISH Detection of X and Y Chromosomes in Combination with Immunofluorescence to Study Contribution of Transplanted Cells to Skeletal Muscle Fibers.

    Science.gov (United States)

    Strömberg, Anna; Jansson, Monika

    2015-01-01

    During the past decades, several studies in animals have displayed the ability of cells from the bone marrow (BM) to participate in regeneration of various tissues including skeletal muscle tissue. Studies in mice have demonstrated that regular physical activity is sufficient to induce contribution of BM derived cells to the skeletal muscle tissue, suggesting that this is part of the physiological remodeling of skeletal muscle. To analyze whether BM-derived cells participate in skeletal muscle remodeling in human, we developed a protocol of immunofluorescence in combination with fluorescence in situ hybridization (FISH) that enables the detection of male donor bone marrow cell contribution to female skeletal muscle tissue.

  12. [Pregnancy-specific dermatoses].

    Science.gov (United States)

    Soutou, B; Aractingi, S

    2015-03-01

    Pregnancy-specific dermatoses include polymorphic eruption of pregnancy, atopic eczema of pregnancy, and pemphigoid gestationis. Intrahepatic cholestasis of pregnancy and impetigo herpetiformis are not real pregnancy-specific dermatoses but they are important to know considering the fetal and maternal risks. Polymorphic eruption of pregnancy is a pruritic disease that usually occurs in primiparous women during the last trimester of pregnancy. Atopic eczema of pregnancy is still controversial as an entity covering conditions with eczematous lesions, prurigo, or folliculitis, and inconstantly associated with a personal history of atopy. Skin biopsy with direct immunofluorescence or search for serum anti-BPAg1 (180kD) NC16a antibodies is mandatory in pruritic dermatoses of pregnancy in order to rule out pemphigoid gestationis. Serum bile salts levels should be tested whenever a generalized pruritus develops during pregnancy in order to rule out intrahepatic cholestasis. Copyright © 2014 Société nationale française de médecine interne (SNFMI). Published by Elsevier SAS. All rights reserved.

  13. Epidermolysis bullosa acquisita and anti-p200 pemphigoid as major subepidermal autoimmune bullous diseases diagnosed by floor binding on indirect immunofluorescence microscopy using human salt-split skin

    Directory of Open Access Journals (Sweden)

    Nupur Goyal

    2017-01-01

    Conclusion: In this study, we report three cases of anti-p200 pemphigoid from India. These cases, though indistinguishable clinically from bullous pemphigoid, revealed a floor-binding pattern on indirect immunofluorescence using salt-split skin.

  14. Incomplete nuclear transformation of human spermatozoa in oligo-astheno-teratospermia: characterization by indirect immunofluorescence of chromatin and thiol status.

    NARCIS (Netherlands)

    Ramos, L.; Heijden, GW van der; Derijck, A.; Berden, J.H.M.; Kremer, J.A.; Vlag, J. van der; Boer, P. de

    2008-01-01

    BACKGROUND Sperm heterogeneity in the human, as observed in oligo-astheno-teratozoospermia (OAT), is associated with hypospermatogenesis. METHODS The chromatin of sperm from OAT and normospermic males was characterized with antibodies specific for nucleosomes, the histone H3.1/H3.2 isoform, histone

  15. Comparison of slot blot nucleic acid hybridization, immunofluorescence, and virus isolation techniques to detect bluetongue virus in blood mononuclear cells from cattle with experimentally induced infection.

    Science.gov (United States)

    de la Concha-Bermejillo, A; Schore, C E; Dangler, C A; de Mattos, C C; de Mattos, C A; Osburn, B I

    1992-12-01

    A slot blot hybridization technique was applied for detection of bluetongue virus (BTV) in blood mononuclear cells (BMNC) obtained from cattle with experimentally induced infection. This technique lacked sensitivity to detect the viral nucleic acid directly in clinical specimens. When aliquots of mononuclear cells from these cattle were cultivated in vitro for 10 days to amplify virus titer, only 33.3% of the samples collected during viremia gave a positive signal in the slot blot hybridization format. By contrast, results for 34.3% of noncultured and 63.3% of cultured mononuclear cell samples collected during viremia were positive by immunofluorescence. The average number of infected cells, as detected by immunofluorescence in the noncultured mononuclear cell samples, was 1 to 5/300,000, and was usually > 10/300,000 in the cultured cell samples. Virus was isolated from all postinoculation blood samples obtained from 4 heifers that were seronegative at the time of inoculation, but was not isolated from any of the preinoculation samples, or from any of the postinoculation samples obtained from 2 heifers that were seropositive at the time of inoculation. When virus isolation was attempted from separated mononuclear cells in 2 heifers, 43.7% of the noncultured and 87.5% of the cultured samples had positive results.

  16. Eucaryotic expression of the nucleocapsid protein gene of porcine circovirus type 2 and use of the protein in an indirect immunofluorescence assay for serological diagnosis of postweaning multisystemic wasting syndrome in pigs.

    Science.gov (United States)

    Racine, Sébastien; Kheyar, Ali; Gagnon, Carl A; Charbonneau, Benoît; Dea, Serge

    2004-07-01

    The purpose of this study was to develop a sensitive, rapid, and inexpensive immunofluorescence assay (IFA) using a recombinant porcine circovirus type 2 (PCV2) nucleocapsid protein for the serological detection of PCV2-specific antibodies in pig sera. The viral nucleocapsid protein encoded by the PCV2 ORF2 gene has recently been identified as the most immunoreactive viral protein that carries type-specific antigenic determinants. The ORF2 sequence of the IAF-2897 strain of PCV2 has been cloned into a pCEP5 eucaryotic expression vector under the control of the cytomegalovirus promoter, downstream of a polyhistidine sequence tag. The recombinant plasmid was used in transfection experiments with human epithelial kidney 293 cells that were further tested, and positive expression of the viral nucleocapsid protein was confirmed by IFA and Western blotting. Strong, specific fluorescence was observed in the nuclei of transfected cells. Test specificity to PCV2 was verified with several related infectious agents. Sensitivity was compared to that of standard IFA using PCV2-infected cells by evaluating the reactivities of 44 field serum samples from pigs on farms with a porcine population suffering from postweaning multisystemic wasting syndrome. The recombinant nucleocapsid-based test was able to detect 15 more positive-testing pigs than the PCV2-based IFA. Therefore, the relative sensitivity of the latter test was estimated at only 57.1% compared to that of the recombinant nucleocapsid-based test. The recombinant fusion protein has been purified by affinity chromatography and is being used to develop further sensitive serological tests.

  17. Preparation of uranium compounds

    Science.gov (United States)

    Kiplinger, Jaqueline L; Montreal, Marisa J; Thomson, Robert K; Cantat, Thibault; Travia, Nicholas E

    2013-02-19

    UI.sub.3(1,4-dioxane).sub.1.5 and UI.sub.4(1,4-dioxane).sub.2, were synthesized in high yield by reacting turnings of elemental uranium with iodine dissolved in 1,4-dioxane under mild conditions. These molecular compounds of uranium are thermally stable and excellent precursor materials for synthesizing other molecular compounds of uranium including alkoxide, amide, organometallic, and halide compounds.

  18. Bulk and compound-specific isotope analysis of long-chain n-alkanes from a 85,000 year sediment core from Lake Peten Petén Itzá, Guatemala

    Science.gov (United States)

    Mays, J.; Brenner, M.; Curtis, J. H.; Curtis, K.; Hodell, D. A.; Correa-Metrio, A.; Escobar, J.; Dutton, A. L.; Zimmerman, A. R.; Guilderson, T. P.

    2013-12-01

    Sediment core PI-6 from Lake Petén Itzá, Guatemala possesses an 85-ka record of climate from lowland Central America. Variations in sediment lithology suggest large, abrupt changes in precipitation during the last glacial and deglacial periods, and into the early Holocene. Study of cores from nearby Lake Quexil demonstrated the utility of using the carbon isotopic composition of leaf wax n-alkanes to infer changes in terrestrial vegetation (Huang et al. 2001). Forty-nine samples were taken from composite Petén Itzá core PI-6 to measure carbon isotopes of bulk organic carbon and long-chain n alkanes. Changes in δ13C values indicate shifts in the relative proportion of C3 to C4 biomass. The record shows largest δ13C variations are associated with Heinrich Events. Carbon isotope values in sediments deposited during the Last Glacial Maximum (LGM) indicate moderate precipitation and little rainfall fluctuation. The deglacial was a period of pronounced climate variability, e.g. the Bölling-Allerod and Younger Dryas. Arid times of the deglacial were inferred from samples with the greatest δ13C values in organic matter, reflecting the largest proportion of C4 plants. Such inferences are supported by stable isotope measurements on ostracod shells and analysis of pollen from the same sample depths in core PI-6. Carbon stable isotope measures on bulk organic carbon and n alkane compounds show similar trends throughout the record and the C:N ratio of Petén Itzá sediments indicates a predominantly allochthonous source for bulk organic matter. Hence, isotope measures on bulk organic carbon (δ13CTOC) in sediments from this lake are sufficient to infer climate-driven shifts in vegetation, making n-alkane extraction and isotope analysis superfluous.

  19. Thiosemicarbazones and Phthalyl-Thiazoles compounds exert antiviral activity against yellow fever virus and Saint Louis encephalitis virus.

    Science.gov (United States)

    Pacca, Carolina Colombelli; Marques, Rafael Elias; Espindola, José Wanderlan P; Filho, Gevânio B O Oliveira; Leite, Ana Cristina Lima; Teixeira, Mauro Martins; Nogueira, Mauricio L

    2017-03-01

    Arboviruses, arthropod-borneviruses, are frequency associated to human outbreak and represent a serious health problem. The genus Flavivirus, such as Yellow Fever Virus (YFV) and Saint Louis Encephalitis Virus (SLEV), are important pathogens with high morbidity and mortality worldwide. In Brazil, YFV is maintained in sylvatic cycle, but many cases are notified annually, despite the efficiency of vaccine. SLEV causes an acute encephalitis and is widely distributed in the Americas. There is no specific antiviral drugs for these viruses, only supporting treatment that can alleviate symptoms and prevent complications. Here, we evaluated the potential anti-YFV and SLEV activity of a series of thiosemicarbazones and phthalyl-thiazoles. Plaque reduction assay, flow cytometry, immunofluorescence and cellular viability were used to test the compounds in vitro. Treated cells showed efficient inhibition of the viral replication at concentrations that presented minimal toxicity to cells. The assays showed that phthalyl-thiazole and phenoxymethyl-thiosemicarbazone reduced 60% of YFV replication and 75% of SLEV replication. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

  20. Simple sensitive rapid detection of Escherichia coli O157:H7 in food samples by label-free immunofluorescence strip sensor.

    Science.gov (United States)

    Song, Chunmei; Li, Jianwu; Liu, Jinxin; Liu, Qing

    2016-08-15

    A simple, one-step, rapid method to detect Escherichia coli O157: H7 (E. coli O157: H7) using a label-free immunofluorescence strip sensor is presented. Fluorescein isothiocyanate (FITC) was added to the sample culture medium to prepare the fluorescent probe for the label-free strip sensor. With the presence of E. coli O157: H7 in the samples, the bacteria could emit a yellow-green fluorescence after incubation and maintain good affinity to the monoclonal antibodies (McAb) against E. coli O157: H7. The direct-type immunofluorescence strip sensor was based on the binding between fluorescent bacteria and the unlabeled McAb immobilized at the test line in nitrocellulose membrane (NC membrane) reaction zone. The visual limit of detection (LOD) of the strip for qualitative detection was 10(6)cells/mL while the LOD for semi-quantitative detection could go down to 10(5)cells/mL by using scanning reader. The LOD was substantially improved to 1cells/mL of the original bacterial content after pre-incubation of the bread, milk and jelly samples in broth for 10, 10 and 8h respectively, which was competitive to some current rapid E. coli O157: H7 detection methods. Besides the obvious advantages, including reduced detection time and operation procedures, the results of this method meet the various detection requirements for E. coli O157: H7 and are comparable to the traditional enzyme-linked immunosorbent assay (ELISA) and double antibody sandwich gold-labeled strips. This is the first report of semi-quantitative immunofluorescence strip for directly detecting foodborne pathogen using only one unlabeled antibody. All detections could be achieved in less than 5min. In addition, this simple, low-cost and easy to be popularized method served as a significant step towards the development of monitoring foodborne pathogens in food-safety testing. Copyright © 2016 Elsevier B.V. All rights reserved.

  1. Clinical significance of positive immunoblotting but negative immunofluorescence for antimitochondrial antibodies in patients with liver diseases other than primary biliary cirrhosis.

    Science.gov (United States)

    Masuda, Jun-Ichi; Omagar, Katsuhisa; Miyakawa, Hiroshi; Hazama, Hiroaki; Ohba, Kazuo; Kinoshita, Hideki; Matsuo, Isao; Isomoto, Hajime; Murata, Ikuo; Kohno, Shigeru

    2002-03-01

    The serum reaction to anti-2-oxo-acid dehydrogenase complex (2-OADC) enzymes, the antigens recognized by antimitochondrial antibodies (AMA), can be detected by immunoblotting in patients with liver diseases other than primary biliary cirrhosis (PBC), who are negative for AMA by conventional indirect immunofluorescence. Whether the presence of anti-2-OADC is related to PBC or represents preclinical PBC in such patients is obscure at present. We examined the immunoreactivity of AMA by immunofluorescense, immunoblotting, and enzyme inhibition assay in serum samples from 59 patients with liver diseases other than PBC and 71 healthy subjects. We also examined the clinical course of the patients in whom a positive result was obtained to elucidate whether such reaction was a "true" or "false" phenomenon. None of the 130 sera was positive for AMA by indirect immunofluorescence or for anti-pyruvate dehydrogenase complex (PDC) by enzyme inhibition assay. However, seven of 71 (10%) sera from healthy subjects contained weak IgG class antibody to PDC-E2 (four sera) or E2 subunit of branched-chain oxo-acid dehydrogenase complex (BCOADC-E2) (three sera). Of the 59 sera from patients with liver diseases other than PBC, four (7%) reacted against 2-OADC by immunoblotting. Of these, three sera were from patients with chronic hepatitis C virus (HCV) infection, and contained IgG class autoantibody to BCOADC-E2. The serum reactivity to BCOADC-E2 detected by immunoblotting in these three patients diminished after absorption with recombinant BCOADC-E2 fusion protein. During the 3-5 year follow-up period, AMA by immunofluorescence and anti-PDC activity by enzyme inhibition assay were always negative in these three patients. The other one serum was from patient with alcoholic cirrhosis, and contained IgM class autoantibody to E3 binding protein (E3-BP). This patient did not develop PBC during the following 2 years. Our results showed that anti-2-OADC antibodies could be detected in some

  2. Coordination Compounds in Biology

    Indian Academy of Sciences (India)

    Home; Journals; Resonance – Journal of Science Education; Volume 4; Issue 6. Coordination Compounds in Biology - The Chemistry of Vitamin B12 and Model Compounds. K Hussian Reddy. General Article Volume 4 Issue 6 June 1999 pp 67-77 ...

  3. 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.

  4. Differentiating antiepiligrin cicatricial pemphigoid from epidermolysis bullosa acquisita by indirect immunofluorescence of skin substrates lacking Type VII collagen or laminin 332: a case report and review of literature

    Directory of Open Access Journals (Sweden)

    Chih-Pin Chen

    2011-06-01

    Full Text Available Antiepiligrin cicatricial pemphigoid (AECP is a chronic autoimmune subepidermal blistering disease characterized by clinical features of cicatricial pemphigoid and circulating IgG antibasement membrane autoantibodies directed against laminin 332. There is growing evidence of an increased relative risk for solid cancers and lymphomas in AECP patients, especially in the 1st year after the onset of blisters. However, it is difficult to distinguish patients with initially skin-predominant AECP from similar findings of epidermolysis bullosa acquisita merely based on clinical, histopathologic, and immuno-pathologic examinations. This is a report on a case of AECP confirmed by indirect immunofluorescence of type VII collagen- and laminin 332-deficient skin as substrates to differentiate it from epidermolysis bullosa acquisita.

  5. [Serologic cross-reactions to Leishmania infantum using indirect immunofluorescence in HIV+ and HIV- patients with active tuberculosis].

    Science.gov (United States)

    López-Vélez, R; Turientes, M C; Gómez-Mampaso, E

    1998-03-01

    Clinical presentation of disseminated tuberculosis and visceral leishmaniosis can be very similar, mainly in those infected with HIV, being serology a useful tool in making a differential diagnosis. Cross-reactions of IFAT serodiagnosis of visceral leishmaniosis with other diseases are well known, but few data is available with tuberculosis. Detection of serum antibodies against Leishmania, using a commercial IFAT kit, was attempted in sera of 51 patients with active pulmonar and/or extrapulmonar tuberculosis (25 HIV+ and 26 HIV-). Overall cross-reactions was found in 19.6% patients without significative differences in between 2 groups, but differences in positive serum titres was observed: one at 1/256, three at 1/160, and one at 1/80 dilution, in the HIV+ group, whereas all 5 patients in HIV- group cross-reacted at 1/80 dilution. Recognition of specific leishmanial antigenic bands by serum antibodies of patients with tuberculosis were not clearly defined by Western-blot. IFAT technique for leishmaniosis cross-react in 20% of patients with tuberculosis.

  6. Immunofluorescence reveals unusual patterns of labelling for connexin43 localized to calbindin-D28K-positive interstitial cells in the pineal gland.

    Science.gov (United States)

    Tsao, D D; Wang, S G; Lynn, B D; Nagy, J I

    2017-06-01

    Gap junctions between cells in the pineal gland have been described ultrastructurally, but their connexin constituents have not been fully characterized. We used immunofluorescence in combination with markers of pineal cells to document the cellular localization of connexin43 (Cx43). Immunofluorescence labelling of Cx43 with several different antibodies was widely distributed throughout the pineal, whereas another connexin examined, connexin26, was not found in pineal but only in surrounding leptomeninges. Labelling apparently associated with plasma membranes was visualized either as fine Cx43-puncta (1-2 μm) or as unusually large pools of Cx43 ranging up to 4-7 μm in diameter or length. These puncta and pools were highly concentrated in perivascular spaces, where they were associated with numerous cells devoid of labelling for markers of pinealocytes (e.g. tryptophan hydroxylase and serotonin), and where they were minimally associated with blood vessels and lacked association with resident macrophages. Astrocytes labelled for glial fibrillary acidic protein were largely restricted to the anterior pole of the pineal gland, where they displayed only fine and sparse Cx43-puncta along their processes. Labelling for Cx43 was localized largely though not exclusively to the somata and long processes of a subpopulation of perivascular interstitial cells that were immunopositive for calbindin-D28K. These cells were often located among dense bundles or termination areas of sympathetic fibres labelled for tyrosine hydroxylase or serotonin. The results indicate that interstitial cells form abundant gap junctions composed of Cx43, and suggest that gap junction-mediated intracellular communication by these cells supports the activities of pinealocytes. © 2017 Federation of European Neuroscience Societies and John Wiley & Sons Ltd.

  7. Production of Monoclonal Antibodies Against the Challenge Strain of Infectious Laryngotracheitis Virus of Chickens and Their Use in an Indirect Immunofluorescent Diagnostic Test

    Directory of Open Access Journals (Sweden)

    Ferhat Abbas*, James Andreasen1, Rockey Becker1, Masroor Ahmed, M Arif Awan, Abdul Wadood and Anita Sonn1

    2010-07-01

    Full Text Available The objective of the present research was to produce monoclonal antibodies (MCAs against the USDA challenge strain of infectious laryngotracheitis virus and to perform an initial investigation of their use in an indirect immunofluorescence diagnostic test. Fourteen-day old chicken embryo liver cells were grown in tissue culture plates. Confluent monolayers were obtained after 48 hours. Monolayers were infected with the USDA challenge strain of infectious laryngotracheitis virus (ILTV. Cytopethic effect of the virus in the form of syncytial formation and clumping of cells was observed after 24 hours. The virus from the tissue culture flasks was collected and purified using discontinuous sucrose gradient. A clear band of the virus from sucrose gradient was obtained. The refractory index and the density measured were 1.410 and 1.20 g/cm3, respectively. Spectrophotometry of the purified virus showed 68.117 ug/ml of protein and 9.8948 ug/ml of nucleic acid concentration. Spleen cells from immunized mice with pure virus were fused with myeloma cells and hybridomas were obtained after 10 days. Screening was performed using indirect immunofluorescence antibody test (IFAT using rabbit anti-mouse immunoglobulins as secondary antibodies. Three hybridomas, 2D1D8, 2E11G2 and 2C6C7 were found producing antibodies against ILTV. All monoclonal antibodies were of isotype IgM and reacted with different strains of ILTV (ILTV USDA, S 88 00224, 86-1169 in IFAT. None of the monoclonals reacted with Parrot herpesvirus and avian adenovirus 301 in IFAT.

  8. 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

  9. Specific Phobias

    Science.gov (United States)

    ... Health This information in Spanish ( en español ) Specific phobias Treatment More information on specific phobias A specific ... psychotherapy. Return to top More information on Specific phobias Explore other publications and websites Phobias (Copyright © Mayo ...

  10. High-resolution view of compound promiscuity.

    Science.gov (United States)

    Hu, Ye; Bajorath, Jürgen

    2013-01-01

    Compound promiscuity is defined as the ability of a small molecule to specifically interact with multiple biological targets. So-defined promiscuity is relevant for drug discovery because it provides the molecular basis of polypharmacology, which is increasingly implicated in the therapeutic efficacy of drugs. Recent studies have analyzed different aspects of compound promiscuity on the basis of currently available activity data. In this commentary, we present take-home messages from these studies augmented with new results to generate a detailed picture of compound promiscuity that might serve as a reference for further discussions and research activities.

  11. W3CoB3-type {Y, Gd - Ho}3Co4-xAlx (x=0.5-1) rare earth compounds: Specific features of crystal structure and magnetic ordering

    Science.gov (United States)

    Morozkin, A. V.; Garshev, A. V.; Knotko, A. V.; Yapaskurt, V. O.; Isnard, O.; Yao, Jinlei; Nirmala, R.; Quezado, S.; Malik, S. K.

    2017-07-01

    The crystal structure of new W3CoB3-type {Y, Gd - Ho}3Co3.25Al0.75, Gd3Co3.5Al0.5 and Tb3Co3Al compounds (Cmcm. N 63, oC28) has been established using powder X-ray diffraction studies. The magnetic properties of Gd3Co3.5Al0.5, Gd3Co3.25Al0.75 and Tb3Co3.25Al0.75 were determined by bulk magnetization measurements and neutron diffraction studies. Gd3Co0.5Al0.5, Gd3Co3.25Al0.75 and Tb3Co3.25Al0.75 exhibit ferrimagnetic ordering below 196 K, 161 K and 151 K, respectively. Tb3Co3.25Al0.75 shows a spin-reorientation transition at 42 K. Below the ferrimagnetic ordering temperature Gd3Co3.25Al0.75 and Tb3Co3.25Al0.75 are soft ferrimagnets, meanwhile Tb3Co3.25Al0.75 shows magnetic hardness below the spin-reorientation transition with remanent magnetization per formula unit of 9.7 μB and coercive field of 15 kOe at 2 K. The magnetocaloric effects of Gd3Co3.25Al0.75 and Tb3Co3.25Al0.75 were calculated in terms of isothermal magnetic entropy change and they reach maximum values of -4.9 J/kg·K at 135-145 K and -3.7 J/kg·K at 115-135 K, respectively, for a field change of 0-50 kOe. Low temperature magnetic ordering in Tb3Co3.25Al0.75 is accompanied by a positive magnetocaloric effect with isothermal magnetic entropy changes of +13.6 J/kg·K at 10 K for a field change of 0-50 kOe and +0.9 J/kg K at 45 K for a field change of 0-10 kOe. Neutron diffraction study in zero applied field shows mixed ferro-antiferromagnetic ordering of Tb3Co3.25Al0.75 with a wave vector K0=[0, 0, 0]. Below 137 K Tb3Co3.25Al0.75 exhibits non-collinear ferrimagnetic ordering of terbium and cobalt sublattices with resulting of b-axis ferromagnetic and c-axis antiferromagnetic components of Cmc‧m={1, mx}×{1, mz/[0, 0, 1/2]}×{1, i‧}×{1, 1/[1/2, 1/2, 0]} magnetic space group. The spin-reorientation transition in Tb3Co3.25Al0.75 below 38 K corresponds to appearance of additional a-axis ferromagnetic component and decreasing of symmetry of magnetic ordering down to C2‧/c={1, mz/[0, 0, 1/2]}×{1, i

  12. Heterogeneous photocatalytic reactions of sulfur aromatic compounds.

    Science.gov (United States)

    Samokhvalov, Alexander

    2011-11-18

    Sulfur aromatic compounds, such as mono-, di-, tri-, and tetraalkyl-substituted thiophene, benzothiophenes, dibenzothiophenes, are the molecular components of many fossils (petroleum, oil shale, tar sands, bitumen). Structural units of natural, cross-linked heteroaromatic polymers present in brown coals, turf, and soil are similar to those of sulfur aromatic compounds. Many sulfur aromatic compounds are found in the streams of petroleum refining and upgrading (naphthas, gas oils) and in the consumer products (gasoline, diesel, jet fuels, heating fuels). Besides fossils, the structural fragments of sulfur aromatic compounds are present in molecules of certain organic semiconductors, pesticides, small molecule drugs, and in certain biomolecules present in human body (pheomelanin pigments). Photocatalysis is the frontier area of physical chemistry that studies chemical reactions initiated by absorption of photons by photocatalysts, that is, upon electronic rather than thermal activation, under "green" ambient conditions. This review provides systematization and critical review of the fundamental chemical and physicochemical information on heterogeneous photocatalysis of sulfur aromatic compounds accumulated in the last 20-30 years. Specifically, the following topics are covered: physicochemical properties of sulfur aromatic compounds, major classes of heterogeneous photocatalysts, mechanisms and reactive intermediates of photocatalytic reactions of sulfur aromatic compounds, and the selectivity of these reactions. Quantum chemical calculations of properties and structures of sulfur aromatic compounds, their reactive intermediates, and the structure of adsorption complexes formed on the surface of the photocatalysts are also discussed. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. Compound Semiconductor Radiation Detectors

    CERN Document Server

    Owens, Alan

    2012-01-01

    Although elemental semiconductors such as silicon and germanium are standard for energy dispersive spectroscopy in the laboratory, their use for an increasing range of applications is becoming marginalized by their physical limitations, namely the need for ancillary cooling, their modest stopping powers, and radiation intolerance. Compound semiconductors, on the other hand, encompass such a wide range of physical and electronic properties that they have become viable competitors in a number of applications. Compound Semiconductor Radiation Detectors is a consolidated source of information on all aspects of the use of compound semiconductors for radiation detection and measurement. Serious Competitors to Germanium and Silicon Radiation Detectors Wide-gap compound semiconductors offer the ability to operate in a range of hostile thermal and radiation environments while still maintaining sub-keV spectral resolution at X-ray wavelengths. Narrow-gap materials offer the potential of exceeding the spectral resolutio...

  14. MEA 86 Compound data

    Data.gov (United States)

    U.S. Environmental Protection Agency — This data file contains the full raw parameter data for the 86 compounds tested in the developmental MEA assay, as well as Area Under the Curve (AUC) calculations...

  15. Volatile organic compound sensor system

    Science.gov (United States)

    Schabron, John F.; Rovani, Jr., Joseph F.; Bomstad, Theresa M.; Sorini-Wong, Susan S.; Wong, Gregory K.

    2011-03-01

    Generally, this invention relates to the development of field monitoring methodology for new substances and sensing chemical warfare agents (CWAs) and terrorist substances. It also relates to a portable test kit which may be utilized to measure concentrations of halogenated volatile organic compounds (VOCs) in the field. Specifically it relates to systems for reliably field sensing the potential presence of such items while also distinguishing them from other elements potentially present. It also relates to overall systems and processes for sensing, reacting, and responding to an indicated presence of such substance, including modifications of existing halogenated sensors and arrayed sensing systems and methods.

  16. Volatile organic compound sensor system

    Science.gov (United States)

    Schabron, John F [Laramie, WY; Rovani, Jr., Joseph F.; Bomstad, Theresa M [Laramie, WY; Sorini-Wong, Susan S [Laramie, WY

    2009-02-10

    Generally, this invention relates to the development of field monitoring methodology for new substances and sensing chemical warfare agents (CWAs) and terrorist substances. It also relates to a portable test kit which may be utilized to measure concentrations of halogenated volatile organic compounds (VOCs) in the field. Specifically it relates to systems for reliably field sensing the potential presence of such items while also distinguishing them from other elements potentially present. It also relates to overall systems and processes for sensing, reacting, and responding to an indicated presence of such substance, including modifications of existing halogenated sensors and arrayed sensing systems and methods.

  17. Chemistry of peroxide compounds

    Science.gov (United States)

    Volnov, I. I.

    1981-01-01

    The history of Soviet research from 1866 to 1967 on peroxide compounds is reviewed. This research dealt mainly with peroxide kinetics, reactivity and characteristics, peroxide production processes, and more recently with superoxides and ozonides and emphasis on the higher oxides of group 1 and 2 elements. Solid state fluidized bed synthesis and production of high purity products based on the relative solubilities of the initial, intermediate, and final compounds and elements in liquid ammonia are discussed.

  18. Phenolic compounds in flaxseed

    OpenAIRE

    Johnsson, Pernilla

    2004-01-01

    The dietary lignan secoisolariciresinol diglucoside (SDG), present in high concentrations in flaxseed, and its metabolites enterolactone and enterodiol are thought to decrease the risk of hormone dependent cancers, cardiovascular disease and other “welfare” diseases. Flaxseed also contains other biologically active phenolic compounds, such as phenolic acids. The understanding of the nature of these compounds is crucial for their possible exploitation in drugs and functional foods. Until the m...

  19. Biodegradation of Organofluorine Compounds

    Science.gov (United States)

    2016-02-01

    3.1 Enrichment Cultures One approach to the identification of degradative enzymes is to “feed” the compound of interest (COI) to a...Defluorination of Organofluorine Sulfur Compounds by Pseudomonas Sp. Strain D2. Environ. Sci. Technol. 1998, 32, 2283–2287. 2. Chan, P.W.Y.; Yakunin...document format ( pdf ) electronic version of this report: U.S. Army Edgewood Chemical Biological Center RDCB-DRB-C ATTN: Dixon, M

  20. Nomenclature of chemical compounds

    OpenAIRE

    Karolina Kaczmarek

    2009-01-01

    This article presents the mechanisms of the inorganic chemistry nomenclature formation in French language. It shows the structure and the way of presenting the names of chemical compounds either descriptively or by giving the structural formulas’ characteristics, their transcription and order of reading the letters. The text specifies the rules of naming a chemical compound, according to the criteria of IUPAC (Union of Pure and Applied Chemistry), the organisation responsible for digesting th...

  1. Phenolic Molding Compounds

    Science.gov (United States)

    Koizumi, Koji; Charles, Ted; de Keyser, Hendrik

    Phenolic Molding Compounds continue to exhibit well balanced properties such as heat resistance, chemical resistance, dimensional stability, and creep resistance. They are widely applied in electrical, appliance, small engine, commutator, and automotive applications. As the focus of the automotive industry is weight reduction for greater fuel efficiency, phenolic molding compounds become appealing alternatives to metals. Current market volumes and trends, formulation components and its impact on properties, and a review of common manufacturing methods are presented. Molding processes as well as unique advanced techniques such as high temperature molding, live sprue, and injection/compression technique provide additional benefits in improving the performance characterisitics of phenolic molding compounds. Of special interest are descriptions of some of the latest innovations in automotive components, such as the phenolic intake manifold and valve block for dual clutch transmissions. The chapter also characterizes the most recent developments in new materials, including long glass phenolic molding compounds and carbon fiber reinforced phenolic molding compounds exhibiting a 10-20-fold increase in Charpy impact strength when compared to short fiber filled materials. The role of fatigue testing and fatigue fracture behavior presents some insight into long-term reliability and durability of glass-filled phenolic molding compounds. A section on new technology outlines the important factors to consider in modeling phenolic parts by finite element analysis and flow simulation.

  2. Highly trifluoromethylated platinum compounds.

    Science.gov (United States)

    Martínez-Salvador, Sonia; Forniés, Juan; Martín, Antonio; Menjón, Babil

    2011-07-11

    The homoleptic, square-planar organoplatinum(II) compound [NBu(4)](2) [Pt(CF(3))(4)] (1) undergoes oxidative addition of CF(3) I under mild conditions to give rise to the octahedral organoplatinum(IV) complex [NBu(4)](2) [Pt(CF(3))(5)I] (2). This highly trifluoromethylated species reacts with Ag(+) salts of weakly coordinating anions in Me(2)CO under a wet-air stream to afford the aquo derivative [NBu(4)][Pt(CF(3))(5) (OH(2))] (4) in around 75% yield. When the reaction of 2 with the same Ag(+) salts is carried out in MeCN, the solvento compound [NBu(4) ][Pt(CF(3))(5)(NCMe)] (5) is obtained in around 80% yield. The aquo ligand in 4 as well as the MeCN ligand in 5 are labile and can be cleanly replaced by neutral and anionic ligands to furnish a series of pentakis(trifluoromethyl)platinate(IV) compounds with formulae [NBu(4)][Pt(CF(3))(5) (L)] (L=CO (6), pyridine (py; 7), tetrahydrothiophene (tht; 8)) and [NBu(4)](2) [Pt(CF(3))(5)X] (X=Cl (9), Br (10)). The unusual carbonyl-platinum(IV) derivative [NBu(4)][Pt(CF(3))(5) (CO)] (6) is thermally stable and has a ν(CO) of 2194 cm(-1). The crystal structures of 2⋅CH(2)Cl(2), 5, [PPh(4) ][Pt(CF(3))(5)(CO)] (6'), and 7 have been established by X-ray diffraction methods. Compound 2 has shown itself to be a convenient entry to the chemistry of highly trifluoromethylated platinum compounds. To the best of our knowledge, compounds 2 and 4-10 are the organoelement compounds with the highest CF(3) content to have been isolated and adequately characterized to date. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  3. 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.

  4. Nomenclature of chemical compounds

    Directory of Open Access Journals (Sweden)

    Karolina Kaczmarek

    2009-01-01

    Full Text Available This article presents the mechanisms of the inorganic chemistry nomenclature formation in French language. It shows the structure and the way of presenting the names of chemical compounds either descriptively or by giving the structural formulas’ characteristics, their transcription and order of reading the letters. The text specifies the rules of naming a chemical compound, according to the criteria of IUPAC (Union of Pure and Applied Chemistry, the organisation responsible for digesting the chemical nomenclature. The article contains the transcription chart and the manner of reading the structural formula, also called latero-numerical. Additionally, there is information conceming the usage of the common names given, still remaining in use next to the names compatible with those of IUPAC. Particular types of chemical compounds have served as models for description of other nomenclature formation rules from the simplest structures to the complicated compound ones. A short summary presents the relations and similarities between the names of particular types of chemical compounds.

  5. Neoclassical compounds and final combining forms in English

    Directory of Open Access Journals (Sweden)

    Ana Díaz-Negrillo

    2014-12-01

    Full Text Available English neoclassical compounds rely on a distinct vocabulary stock and present morphological features which raise a number of theoretical questions. Generalisations about neoclassical compounds are also problematic because the output is by no means homogeneous, that is, defining features of neoclassical compounds sometimes co-exist with features that are not prototypical of these formations. The paper looks at neoclassical compounds with a view to exploring patterns of morphological behaviour and development in this class of compounds. The approach is both synchronic and diachronic: it researches whether the morphological behaviour of recently formed compounds is different from that of earlier compounds and, if so, in which respects. This is assessed on data from the BNC with respect to some of the features that are cited in the literature as defining properties of neoclassical compounds, specifically, their internal configuration, the occurrence or not of a linking vowel, and their productivity.

  6. A retrospective study comparing agar plate culture, indirect immunofluorescence and real-time PCR for the diagnosis of Strongyloides stercoralis infection.

    Science.gov (United States)

    Buonfrate, Dora; Perandin, Francesca; Formenti, Fabio; Bisoffi, Zeno

    2017-05-01

    Strongyloides stercoralis is a parasite that can cause death in immunocompromised people. A proper diagnosis is hence essential. The real-time polymerase-chain reaction (RT-PCR) is a novel, promising diagnostic method, that detects the DNA of the parasite in stool samples. In this retrospective study, we compared the sensitivity of agar plate coproculture (APC), an in-house immunofluorescence test (IFAT) and an in-house RT-PCR for the diagnosis of S. stercoralis infection. The study sample was composed by 223 samples. Samples resulting positive to APC, IFAT and RT-PCR were 20, 140 and 25, respectively. When sensitivity was calculated against a composite reference standard, serology confirmed the best performance (sensitivity 95%), followed by RT-PCR (57%) and APC (45%). In conclusion, in a non-endemic setting, serology is the best screening method, while the combination of APC and RT-PCR does not seem a reasonable approach to increase sensitivity. Both methods can have a role as confirmatory tests for selected cases.

  7. Tyrosine phosphorylation on capacitated human sperm tail detected by immunofluorescence correlates strongly with sperm-zona pellucida (ZP) binding but not with the ZP-induced acrosome reaction.

    Science.gov (United States)

    Liu, D Y; Clarke, G N; Baker, H W G

    2006-04-01

    Protein tyrosine phosphorylation (TP) of human sperm is related to sperm capacitation and zona pellucida (ZP) binding. The aim of this study was to determine whether the TP of capacitated sperm is a useful marker for the ability of sperm to bind to the ZP and undergo the ZP-induced acrosome reaction (AR). Semen samples were obtained from 115 subfertile men with sperm count > or =20 x 10(6)/ml, motility > or =25% and variable morphology. Motile sperm (2 x 10(6)/ml) selected by swim-up were incubated with four oocytes for 2 h, and the number of sperm bound to the ZP and the ZP-induced AR was examined. TP of sperm tail was assessed by immunofluorescence (IF) with anti-phosphotyrosine monoclonal antibody. The time course and effects of dibutyryl cyclic adenosine monophosphate (dbcAMP) and phorbol myristate acetate (PMA) on TP were also studied. TP was stimulated more by dbcAMP (P ZP binding but not with the ZP-induced AR. Sperm TP detected by IF correlates strongly with sperm-ZP binding capacity but not with the ZP-induced AR. This simple IF assay of TP may be a clinically useful test of sperm function that is predictive of normal sperm ZP-binding capacity.

  8. 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.

  9. [Comparison of the indirect immunofluorescence assay performance of Bartonella henselae antigens obtained by co-cultivation in Vero and HeLa cells].

    Science.gov (United States)

    Ergin, Cağrı; Akkaya, Yüksel; Kiriş Satılmış, Ozgün; Yılmaz, Cansev

    2011-07-01

    The laboratory diagnosis of Bartonella henselae infection is mainly based on serological testing by indirect immunofluorescence assay (IFA). Cell line co-cultivation with B.henselae and agar derivated antigens are the two major procedures used for evaluation of anti-Bartonella antibodies. Vero and Hep-2 cell lines are the most commonly used media for co-cultivation both in-house and commercial diagnostic kits production. However, HeLa cells which are easily supplied and grown, also can easily be infected by B.henselae. The aim of this study was to compare the performances of antigens obtained by co-cultivation of B.henselae ATCC 49882 (Houston-1) in Vero and HeLa Cells in IFA serology. Out of 381 sera samples, 127 (33.3%) were found positive and 195 (51.2%) were found negative by IFA performed by both cell line co-cultivations. The total agreement between the methods were found as 84.5% (322/381), and Cohen kappa value was calculated as 0.68 (strong, coherent). As a result, He-La cells were found to be useful for the preparation of B.henselae antigens to be used in IFA for the serologic diagnosis of B.henselae infections. However different genotype strains and cross reactions with other infectious agents should be investigated by further studies before routine applications of HeLa cell co-cultivations procedure is established.

  10. Comparative immunofluorescence and ultrastructural analysis of microtubule organization in Uronema sp., Klebsormidium flaccidum, K. subtilissimum, Stichococcus bacillaris and S. chloranthus (Chlorophyta).

    Science.gov (United States)

    Katsaros, Christos I; Varvarigos, Vasilios; Gachon, Claire M M; Brand, Jerry; Motomura, Taizo; Nagasato, Chikako; Küpper, Frithjof C

    2011-04-01

    A detailed comparative examination of microtubule (MT) organization in interphase and dividing cells of Uronema sp., Klebsormidium flaccidum, K. subtilissimum, Stichococcus bacillaris and S. chloranthus was made using tubulin immunofluorescence and transmission electron microscopy (TEM). During interphase all the species bear a well-organized cortical MT system, consisting of parallel bundles with different orientations. In Uronema sp. the cortical MT bundles are longitudinally oriented, whereas in the other species they are in transverse orientation to the axis of the cells. Considerable differences in MT organization were also observed during stages of mitosis, mainly preprophase, as well as cytokinesis. In Uronema sp., a particular radial MT assembly is organized during preprophase-early prophase, which was not observed in the other species. In Stichococcus a fine MT ring surrounded the nucleus during preprophase and prophase. An MT ring, together with single cytoplasmic MTs, was also found associated with the developing diaphragm during cytokinesis in Stichococcus. A phycoplast participates in cytokinesis in Uronema sp., but not in the other species. In Uronema sp. the centrosome functions as a microtubule organizing center (MTOC) during mitosis, but not during interphase and cytokinesis. The phylogenetic significance of these differences is discussed in combination with SSU/ITS sequencing and other, existing molecular data. Copyright © 2010 Elsevier GmbH. All rights reserved.

  11. Comparison of the Seeplex reverse transcription PCR assay with the R-mix viral culture and immunofluorescence techniques for detection of eight respiratory viruses.

    Science.gov (United States)

    Roh, Kyoung Ho; Kim, Jeeyong; Nam, Myung-Hyun; Yoon, Sooyung; Lee, Chang Kyu; Lee, Kapno; Yoo, Young; Kim, Min Ja; Cho, Yunjung

    2008-01-01

    This study evaluated the clinical usefulness of a newly introduced multiplex reverse transcription PCR assay (Seeplex RV; Seegene, Inc., Seoul, Korea) in patients with respiratory symptoms. Fifty clinical respiratory specimens (45 from children, 5 from adults) were tested for 8 viruses (influenza virus type A and B, parainfluenza virus type 1, 2, 3, respiratory syncytial virus type A and B, and adenovirus) by Seeplex RV (S-RV) and R-mix viral culture with immunofluorescence (VC-IF). Forty (80%) of the 50 samples showed concordant results between S-RV and VC-IF; 24 of these showed the same positive and 16 showed the same negative results. Among the 10 discrepant samples, 9 were S-RV-positive and VC-IF-negative. Six were obtained in patients with lower respiratory tract infection. Only 1 sample was VC-IF-positive and S-RV-negative. This patient had pneumonia. In 3 cases, more than 1 virus was identified by S-RV. The total running time of S-RV was 6 hr, which shortens the detection time for the viral presence by 2 workdays compared to VC-IF. In conclusion, S-RV is reliable, rapid, relatively easy to perform, and able to detect more than 1 virus simultaneously. Therefore, implementation of the S-RV assay in clinical laboratories will aid rapid diagnosis and treatment of major viral infections of the respiratory tract.

  12. Compound semiconductor device physics

    CERN Document Server

    Tiwari, Sandip

    2013-01-01

    This book provides one of the most rigorous treatments of compound semiconductor device physics yet published. A complete understanding of modern devices requires a working knowledge of low-dimensional physics, the use of statistical methods, and the use of one-, two-, and three-dimensional analytical and numerical analysis techniques. With its systematic and detailed**discussion of these topics, this book is ideal for both the researcher and the student. Although the emphasis of this text is on compound semiconductor devices, many of the principles discussed will also be useful to those inter

  13. Synthetic Aperture Compound Imaging

    DEFF Research Database (Denmark)

    Hansen, Jens Munk

    Medical ultrasound imaging is used for many purposes, e.g. for localizing and classifying cysts, lesions, and other processes. Almost any mass is first observed using B-mode imaging and later classified using e.g. color flow, strain, or attenuation imaging. It is therefore important that the B...... region of parenchyma. A successful approach to reduce the speckle artifacts is spatial compounding, where images are acquired from a number of directions and combined after envelope-detection. Today, spatial compounding is implemented in all highend ultrasound systems and available when using a low pitch...

  14. Prediction of intermetallic compounds

    Energy Technology Data Exchange (ETDEWEB)

    Burkhanov, Gennady S; Kiselyova, N N [A A Baikov Institute of Metallurgy and Materials Science, Russian Academy of Sciences, Moscow (Russian Federation)

    2009-06-30

    The problems of predicting not yet synthesized intermetallic compounds are discussed. It is noted that the use of classical physicochemical analysis in the study of multicomponent metallic systems is faced with the complexity of presenting multidimensional phase diagrams. One way of predicting new intermetallics with specified properties is the use of modern processing technology with application of teaching of image recognition by the computer. The algorithms used most often in these methods are briefly considered and the efficiency of their use for predicting new compounds is demonstrated.

  15. Polymeric coordination compounds

    Indian Academy of Sciences (India)

    Administrator

    Metal coordination polymers with one- and two-dimensional structures are of current interest due to their possible relevance to material science 1. In continuation of our previous studies 2,3, several new polymeric compounds are reported here. Among the complexes of silver with aminomethyl pyridine (amp) ...

  16. NATURAL POLYACETYLENE COMPOUNDS

    Directory of Open Access Journals (Sweden)

    A. M. Nasukhova

    2014-01-01

    Full Text Available In article the review of the initial stage of researches of natural polyacetylene compounds is resulted. The high reactionary ability leading to fast oxidation and degradation of these compounds, especially at influence of Uf-light, oxygen of air, pH and other factors, has caused the serious difficulties connected with an establishment of structure and studying of their physical and chemical properties. Therefore the greatest quantity of works of this stage is connected with studying of essential oils of plants from families Apiaceae, Araliaceae, Asteraceae, Campanulaceae, Olacaceae, Pittosporaceae and Santalaceae where have been found out, basically, diacetylene compounds. About development of physical and chemical methods of the analysis of possibility of similar researches have considerably extended. More than 2000 polyacetylenes are known today, from them more than 1100 are found out in plants fam. Asteraceae. Revolution in the field of molecular biology has allowed to study processes of biosynthesis of these compounds intensively.

  17. Toxicology of perfluorinated compounds

    Energy Technology Data Exchange (ETDEWEB)

    Stahl, Thorsten [Hessian State Laboratory, Wiesbaden (Germany); Mattern, Daniela; Brunn, Hubertus [Hessian State Laboratory, Giessen (Germany)

    2011-12-15

    Perfluorinated compounds [PFCs] have found a wide use in industrial products and processes and in a vast array of consumer products. PFCs are molecules made up of carbon chains to which fluorine atoms are bound. Due to the strength of the carbon/fluorine bond, the molecules are chemically very stable and are highly resistant to biological degradation; therefore, they belong to a class of compounds that tend to persist in the environment. These compounds can bioaccumulate and also undergo biomagnification. Within the class of PFC chemicals, perfluorooctanoic acid and perfluorosulphonic acid are generally considered reference substances. Meanwhile, PFCs can be detected almost ubiquitously, e.g., in water, plants, different kinds of foodstuffs, in animals such as fish, birds, in mammals, as well as in human breast milk and blood. PFCs are proposed as a new class of 'persistent organic pollutants'. Numerous publications allude to the negative effects of PFCs on human health. The following review describes both external and internal exposures to PFCs, the toxicokinetics (uptake, distribution, metabolism, excretion), and the toxicodynamics (acute toxicity, subacute and subchronic toxicities, chronic toxicity including carcinogenesis, genotoxicity and epigenetic effects, reproductive and developmental toxicities, neurotoxicity, effects on the endocrine system, immunotoxicity and potential modes of action, combinational effects, and epidemiological studies on perfluorinated compounds). (orig.)

  18. Computing compound distributions faster

    OpenAIRE

    Iseger, P.; Smith, M.A.J.; Dekker, Rommert

    1997-01-01

    textabstractThe use of Panjer's algorithm has meanwhile become a widespread standard technique for actuaries (Kuon et al., 1955). Panjer's recursion formula is used for the evaluation of compound distributions and can be applied to life and general insurance problems. The discrete version of Panjer's recursion formula is often applied to continuous distributions by discretizing the

  19. Computing compound distributions faster

    NARCIS (Netherlands)

    P. den Iseger; M.A.J. Smith; R. Dekker (Rommert)

    1997-01-01

    textabstractThe use of Panjer's algorithm has meanwhile become a widespread standard technique for actuaries (Kuon et al., 1955). Panjer's recursion formula is used for the evaluation of compound distributions and can be applied to life and general insurance problems. The discrete version of

  20. Coordination Compounds in Biology

    Indian Academy of Sciences (India)

    inorganic chemistry has been uncovered while studying sys- tems pertinent to B. 12 ... inorganic chemistry. His research interests are in the chemistry of model compounds of biochemical relevance. He did post- doctoral research in the. University of ..... G N Schrauzer, in Advances in Chemistry series, No. 100, Ed. R F.

  1. Stimulation of Ideas through Compound-Based Bibliometrics: Counting and Mapping Chemical Compounds for Analyzing Research Topics in Chemistry, Physics, and Materials Science.

    Science.gov (United States)

    Barth, Andreas; Marx, Werner

    2012-12-01

    Counting compounds (rather than papers or citations) offers a new perspective for quantitative analyses of research activities. First of all, we can precisely define (compound-related) research topics and access the corresponding publications (scientific papers as well as patents) as a measure of research activity. We can also establish the time evolution of the publications dealing with specific compounds or compound classes. Moreover, the mapping of compounds by establishing compound-based landscapes has some potential to visualize the compound basis of research topics for further research activities. We have analyzed the rare earth compounds to give an example of a broad compound class. We present the number of the currently existing compounds and of the corresponding publications as well as the time evolution of the papers and patents. Furthermore, we have analyzed the rare earth cuprates (copper oxides) as an example of a narrower compound class to demonstrate the potential of mapping compounds by compound-based landscapes. We have quantified the various element combinations of the existing compounds and revealed all element combinations not yet realized in the synthesis within this compound class. Finally, we have analyzed the quasicrystal compound category as an example of a compound class that is not defined by a specific element combination or a molecular structure.

  2. Xenobiotic organic compounds in wastewater

    DEFF Research Database (Denmark)

    Eriksson, Eva; Baun, Anders; Henze, Mogens

    2002-01-01

    Information regarding the contents of xenobiotic organic compounds (XOCs) in wastewater is limited, but it has been shown that at least 900 different compounds / compound groups could potentially be present in grey wastewater. Analyses of Danish grey wastewater revealed the presence of several hu...... aquatic toxicity were present and that data for environmental fate could only be retrieved for about half of the compounds....

  3. (I) supramolecular compounds constructed from pyridinium or ...

    Indian Academy of Sciences (India)

    extensive attention, owing to the amazing accuracy and high efficiency in control and arrangement of specific building blocks.1–4 Supramolecular polymers are con- structed generally via .... Silver (I) complexes constructed from multications. 1519. Figure 6. K-M function versus energy (eV) curve compounds 1, 2 and 3.

  4. Medicinal Uses of Inorganic Compounds - 1

    Indian Academy of Sciences (India)

    Anticancer. Antidiabetic. Anti-HIV activity. Skin ointment. Antiviral. Supplement. Antiperspirant. Synthetic antioxidant. Anti-inflammatory neuroprotective agent. Antihypertensive ... a specific inhibitor of the GSK-3 family of protein kinases in vitro and in intact cells. .... ing, ruthenium compounds interact with proteins including.

  5. 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

  6. Prevalence estimation of tick-borne encephalitis virus (TBEV) antibodies in dogs from Finland using novel dog anti-TBEV IgG MAb-capture and IgG immunofluorescence assays based on recombinant TBEV subviral particles.

    Science.gov (United States)

    Levanov, Lev; Vera, Cristina Pérez; Vapalahti, Olli

    2016-07-01

    Tick-borne encephalitis (TBE) is one of the most dangerous human neurological infections occurring in Europe and Northern parts of Asia with thousands of cases and millions vaccinated against it. The risk of TBE might be assessed through analyses of the samples taken from wildlife or from animals which are in close contact with humans. Dogs have been shown to be a good sentinel species for these studies. Serological assays for diagnosis of TBE in dogs are mainly based on purified and inactivated TBEV antigens. Here we describe novel dog anti-TBEV IgG monoclonal antibody (MAb)-capture assay which is based on TBEV prME subviral particles expressed in mammalian cells from Semliki Forest virus (SFV) replicon as well as IgG immunofluorescence assay (IFA) which is based on Vero E6 cells transfected with the same SFV replicon. We further demonstrate their use in a small-scale TBEV seroprevalence study of dogs representing different regions of Finland. Altogether, 148 dog serum samples were tested by novel assays and results were compared to those obtained with a commercial IgG enzyme immunoassay (EIA), hemagglutination inhibition test and IgG IFA with TBEV infected cells. Compared to reference tests, the sensitivities of the developed assays were 90-100% and the specificities of the two assays were 100%. Analysis of the dog serum samples showed a seroprevalence of 40% on Åland Islands and 6% on Southwestern archipelago of Finland. In conclusion, a specific and sensitive EIA and IFA for the detection of IgG antibodies in canine sera were developed. Based on these assays the seroprevalence of IgG antibodies in dogs from different regions of Finland was assessed and was shown to parallel the known human disease burden as the Southwestern archipelago and Åland Islands in particular had considerable dog TBEV antibody prevalence and represent areas with high risk of TBE for humans. Copyright © 2016 Elsevier GmbH. All rights reserved.

  7. Cross-reactive antibodies in convalescent SARS patients' sera against the emerging novel human coronavirus EMC (2012) by both immunofluorescent and neutralizing antibody tests.

    Science.gov (United States)

    Chan, Kwok-Hung; Chan, Jasper Fuk-Woo; Tse, Herman; Chen, Honglin; Lau, Candy Choi-Yi; Cai, Jian-Piao; Tsang, Alan Ka-Lun; Xiao, Xincai; To, Kelvin Kai-Wang; Lau, Susanna Kar-Pui; Woo, Patrick Chiu-Yat; Zheng, Bo-Jiang; Wang, Ming; Yuen, Kwok-Yung

    2013-08-01

    A severe acute respiratory syndrome (SARS)-like disease due to a novel betacoronavirus, human coronavirus EMC (HCoV-EMC), has emerged recently. HCoV-EMC is phylogenetically closely related to Tylonycteris-bat-coronavirus-HKU4 and Pipistrellus-bat-coronavirus-HKU5 in Hong Kong. We conducted a seroprevalence study on archived sera from 94 game-food animal handlers at a wild life market, 28 SARS patients, and 152 healthy blood donors in Southern China to assess the zoonotic potential and evidence for intrusion of HCoV-EMC and related viruses into humans. Anti-HCoV-EMC and anti-SARS-CoV antibodies were detected using screening indirect immunofluorescence (IF) and confirmatory neutralizing antibody tests. Two (2.1%) animal handlers had IF antibody titer of ≥ 1:20 against both HCoV-EMC and SARS-CoV with neutralizing antibody titer of SARS patients had significant IF antibody titers with 7/28 (25%) having anti-HCoV-EMC neutralizing antibodies at low titers which significantly correlated with that of HCoV-OC43. Bioinformatics analysis demonstrated a significant B-cell epitope overlapping the heptad repeat-2 region of Spike protein. Virulence of SARS-CoV over other betacoronaviruses may boost cross-reactive neutralizing antibodies against other betacoronaviruses. Convalescent SARS sera may contain cross-reactive antibodies against other betacoronaviruses and confound seroprevalence study for HCoV-EMC. Copyright © 2013 The British Infection Association. Published by Elsevier Ltd. All rights reserved.

  8. The inter-observer reading variability in anti-nuclear antibodies indirect (ANA) immunofluorescence test: A multicenter evaluation and a review of the literature.

    Science.gov (United States)

    Rigon, A; Infantino, M; Merone, M; Iannello, G; Tincani, A; Cavazzana, I; Carabellese, N; Radice, A; Manfredi, M; Soda, P; Afeltra, A

    2017-12-01

    Recently there has been an increase demand for Computer-Aided Diagnosis (CAD) tools to support clinicians in the field of Indirect ImmunoFluorescence (IIF), as the novel digital imaging reading approach can help to overcome the reader subjectivity. Nevertheless, a large multicenter evaluation of the inter-observer reading variability in this field is still missing. This work fills this gap as we evaluated 556 consecutive samples, for a total of 1679 images, collected in three laboratories with IIF expertise using HEp-2 cell substrate (MBL) at 1:80 screening dilution according to conventional procedures. In each laboratory, the images were blindly classified by two experts into three intensity classes: positive, negative, and weak positive. Positive and weak positive ANA-IIF results were categorized by the predominant fluorescence pattern among six main classes. Data were pairwise analyzed and the inter-observer reading variability was measured by Cohen's kappa test, revealing a pairwise agreement little further away than substantial both for fluorescence intensity and for staining pattern recognition (k=0.602 and k=0.627, respectively). We also noticed that the inter-observer reading variability decreases when it is measured with respect to a gold standard classification computed on the basis of labels assigned by the three laboratories. These data show that laboratory agreement improves using digital images and comparing each single human evaluation to potential reference data, suggesting that a solid gold standard is essential to properly make use of CAD systems in routine work lab. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. Immunofluorescent analysis of testicular biopsies with germ cell and Sertoli cell markers shows significant MVH negative germ cell depletion with older age at orchiopexy.

    Science.gov (United States)

    Li, Ruili; Thorup, Jorgen; Sun, Cong; Cortes, Dina; Southwell, Bridget; Hutson, John

    2014-02-01

    Undescended testis is the most common defect in male newborns. This condition is associated with increased risks of infertility and testicular malignancy due to abnormal germ cell development in the testes. Early surgery may limit such risks. We analyzed germ cell development vs age at orchiopexy using a germ cell marker and a Sertoli cell marker on testicular biopsies. A total of 22 testicular biopsies at orchiopexy in 20 patients 5 to 24.5 months old were fixed and embedded in paraffin. Sections were processed and labeled with AMH antibody for Sertoli cells and MVH antibody for germ cells for immunofluorescent histochemical analysis. Confocal images were counted using ImageJ (National Institutes of Health, Bethesda, Maryland) for germ cells and testicular tubules. The data were analyzed using linear regression. Sertoli cells were clearly distinguished from MVH positive and negative germ cells located centrally or on basement membranes of tubules. Percentage of tubules with MVH negative germ cells significantly decreased with increasing age at orchiopexy (β = -0.03, p = 0.03). Total tubular numbers and "empty" tubules without germ cells significantly increased with age at orchiopexy (β = 1.15, p = 0.02 and β = 0.44, p = 0.04, respectively). AMH antibody distinguished Sertoli cells from germ cells, and MVH antibody distinguished 2 types of germ cells at different developmental stages. Biopsy at orchiopexy in older patients showed significant germ cell depletion. These results lend support to early surgery to optimize germ cell number. Copyright © 2014 American Urological Association Education and Research, Inc. Published by Elsevier Inc. All rights reserved.

  10. Toxic compounds of Curcuma aeruginosa causes necrosis of mice hepatocytes

    Directory of Open Access Journals (Sweden)

    Eka Pramyrtha Hestianah

    2014-08-01

    Full Text Available BACKGROUND People have been using Curcuma aeruginosa rhizome as a traditional herbal medicine as appetite stimulant, without realizing its side effects. Herbal plants contain tens to hundreds of compounds, some of which are toxic. The aim of this research was to determine which toxic compound of Curcuma aeruginosa rhizome has an impact on apoptosis and PARP-1 expression of hepatocytes in male mice. METHODS Eighty eight male Balb C mice were divided into 10 groups treated respectively with Curcuma aeruginosa rhizome cloroform extract, methanol extract, essential oil, infusion, and press juice, at dosages of 0.004g/kgBW and 0.06g/kgBW, and 1 control group. The treatment was given orally once a day for 10 days and on the 11th day, the research animals were sacrificed, and their liver taken for histopathologic slide preparation with Apopteq Detection Kit, and immunofluorescence. Compounds in Curcuma aeruginosa rhizome were analyzed with gas chromatography/mass spectrometry. The data obtained were analyzed by one-way ANOVA, and Partial Least Squares to determine which compounds had an impact on murine hepatocytes. RESULTS The result of one way ANOVA showed that the chloroform groups at dosages of 0.004g/kgBW and 0.06g/kgBW showed the highest apoptosis of mice’s hepatocytes (p<0.05. There were significant differences in PARP-1 expression between control and treatment groups. The highest PARP-1 expression was in the essential oil group at a dosage of 0.06g/kg BW (p<0.05. CONCLUSION Curcuma aeruginosa rhizome given to mice orally causes necrosis of mice’s hepatocytes.

  11. Toxic compounds of Curcuma aeruginosa causes necrosis of mice hepatocytes

    Directory of Open Access Journals (Sweden)

    Eka Pramyrtha Hestianah

    2015-12-01

    Full Text Available BACKGROUND People have been using Curcuma aeruginosa rhizome as a traditional herbal medicine as appetite stimulant, without realizing its side effects. Herbal plants contain tens to hundreds of compounds, some of which are toxic. The aim of this research was to determine which toxic compound of Curcuma aeruginosa rhizome has an impact on apoptosis and PARP-1 expression of hepatocytes in male mice. METHODS Eighty eight male Balb C mice were divided into 10 groups treated respectively with Curcuma aeruginosa rhizome cloroform extract, methanol extract, essential oil, infusion, and press juice, at dosages of 0.004g/kgBW and 0.06g/kgBW, and 1 control group. The treatment was given orally once a day for 10 days and on the 11th day, the research animals were sacrificed, and their liver taken for histopathologic slide preparation with Apopteq Detection Kit, and immunofluorescence. Compounds in Curcuma aeruginosa rhizome were analyzed with gas chromatography/mass spectrometry. The data obtained were analyzed by one-way ANOVA, and Partial Least Squares to determine which compounds had an impact on murine hepatocytes. RESULTS The result of one way ANOVA showed that the chloroform groups at dosages of 0.004g/kgBW and 0.06g/kgBW showed the highest apoptosis of mice’s hepatocytes (p<0.05. There were significant differences in PARP-1 expression between control and treatment groups. The highest PARP-1 expression was in the essential oil group at a dosage of 0.06g/kg BW (p<0.05. CONCLUSION Curcuma aeruginosa rhizome given to mice orally causes necrosis of mice’s hepatocytes.

  12. Compound semiconductor device modelling

    CERN Document Server

    Miles, Robert

    1993-01-01

    Compound semiconductor devices form the foundation of solid-state microwave and optoelectronic technologies used in many modern communication systems. In common with their low frequency counterparts, these devices are often represented using equivalent circuit models, but it is often necessary to resort to physical models in order to gain insight into the detailed operation of compound semiconductor devices. Many of the earliest physical models were indeed developed to understand the 'unusual' phenomena which occur at high frequencies. Such was the case with the Gunn and IMPATI diodes, which led to an increased interest in using numerical simulation methods. Contemporary devices often have feature sizes so small that they no longer operate within the familiar traditional framework, and hot electron or even quantum­ mechanical models are required. The need for accurate and efficient models suitable for computer aided design has increased with the demand for a wider range of integrated devices for operation at...

  13. Process for compound transformation

    KAUST Repository

    Basset, Jean-Marie

    2016-12-29

    Embodiments of the present disclosure provide for methods of using a catalytic system to chemically transform a compound (e.g., a hydrocarbon). In an embodiment, the method does not employ grafting the catalyst prior to catalysis. In particular, embodiments of the present disclosure provide for a process of hydrocarbon (e.g., C1 to C20 hydrocarbon) metathesis (e.g., alkane, olefin, or alkyne metathesis) transformation, where the process can be conducted without employing grafting prior to catalysis.

  14. Phenolic compounds in oats

    OpenAIRE

    Skoglund, Maria

    2008-01-01

    This research project examined how to treat raw oat material for oat-based food products in order to sustain or increase the levels of phenolic compounds. The focus was mainly on the avenanthramides, which are potentially health beneficial bioactive components found exclusively in oats. A proposed enzymatic decrease in avenanthramide levels when non heat-treated milled oats are steeped in water was investigated. The decrease was strongly suggested to be caused by a polyphenol oxidase. Althoug...

  15. A lubricating compound

    Energy Technology Data Exchange (ETDEWEB)

    Barchan, G.P.; Alekseyenko, V.A.; Bolotnikov, V.S.; Burlov, A.S.; Chigarenko, G.G.; Kogan, V.A.

    1982-01-01

    In a lubricating compound (SK), which contains petroleum or synthetic oil, a complex ether (SE) and an additive (Pr), in order to improve the loading, antifriction and antiwear properties, a complex ether of glycerin of the formula C/sub 3/H/sub 5/R/sub 3/, where R is C/sub 3/H/sub 7/C00, C/sub 17/H/sub 33/C00, is used and 2-(n-tololsulfamino)benzalaniline is used as the additive. The ratio of components in percent is: 2-(n-tololsulfamino)benzalanaline, 0.1 to 0.4; complex ether, 20 to 30 and petroleum or synthetic oil to 100. Oils of different chemical structure and physical and chemical properties are used for making the lubricating compound: vaseline, medicinal, industrial, instrumental MVP, vacuum VM-4 and polyethylsiloxane liquid 132-25. The oil is mixed with the complex ether and additive in the cited proportions with heating to 100 degrees C and intensive mixing. After cooling it is ready for use. The results of tests of the proposed lubricating compound in a facial friction machine for lubricating friction subasemblies of steel and a copper alloy showed significant improvements in properties.

  16. A lubricating compound

    Energy Technology Data Exchange (ETDEWEB)

    Barchan, G.P.; Boltnikov, V.S.; Bulgarevich, A.F.; Chigarenko, G.G.; Ponomarenko, A.G.

    1982-01-01

    In a known lubricating compound (SK) in order to improve the loading, antifriction and antiwear properties, a dicarbonic acid of a complex ether of azelaic acid of the formula (CH/sub 2/)/sub 7/(COOC/sub 2/H/sub 2//sub n+1/)/sub 2/, where n = 4 to 8, is additionally introduced as a complex ether (SE). 1-(2-oxy-1-naphthylazo)-2-naphthol-4-sulfo acid is introduced as an additive. The ratio of components in percent is: 1-(2-oxy-1-naphthylazo)-2-naphthol-4-sulfo acid 0.1 to 0.5 and complex ether, 20 to 30 and petroleum or synthetic oil (Ms) to 100 percent. Synthetic or petroleum oil of varying chemical structure and physical and chemical properties is used to prepare the lubricating compound: industrialnoye-20, vaseline, industrialnoye-50, instrumental MPV, vacuum MV-4 and polytehylsiloxanic liquid 32 to 25. The oil is mixed with the complex ether and the additive in the cited ratios with heating to 100 degrees and intensive mixing. After cooling, an oil ready for use is produced. The lubricating properties of the lubricating compound are studied in a facial friction (Tr) machine with a movable sample of St45.

  17. Antifungal compounds from cyanobacteria.

    Science.gov (United States)

    Shishido, Tânia K; Humisto, Anu; Jokela, Jouni; Liu, Liwei; Wahlsten, Matti; Tamrakar, Anisha; Fewer, David P; Permi, Perttu; Andreote, Ana P D; Fiore, Marli F; Sivonen, Kaarina

    2015-04-13

    Cyanobacteria are photosynthetic prokaryotes found in a range of environments. They are infamous for the production of toxins, as well as bioactive compounds, which exhibit anticancer, antimicrobial and protease inhibition activities. Cyanobacteria produce a broad range of antifungals belonging to structural classes, such as peptides, polyketides and alkaloids. Here, we tested cyanobacteria from a wide variety of environments for antifungal activity. The potent antifungal macrolide scytophycin was detected in Anabaena sp. HAN21/1, Anabaena cf. cylindrica PH133, Nostoc sp. HAN11/1 and Scytonema sp. HAN3/2. To our knowledge, this is the first description of Anabaena strains that produce scytophycins. We detected antifungal glycolipopeptide hassallidin production in Anabaena spp. BIR JV1 and HAN7/1 and in Nostoc spp. 6sf Calc and CENA 219. These strains were isolated from brackish and freshwater samples collected in Brazil, the Czech Republic and Finland. In addition, three cyanobacterial strains, Fischerella sp. CENA 298, Scytonema hofmanni PCC 7110 and Nostoc sp. N107.3, produced unidentified antifungal compounds that warrant further characterization. Interestingly, all of the strains shown to produce antifungal compounds in this study belong to Nostocales or Stigonematales cyanobacterial orders.

  18. Antifungal Compounds from Cyanobacteria

    Directory of Open Access Journals (Sweden)

    Tânia K. Shishido

    2015-04-01

    Full Text Available Cyanobacteria are photosynthetic prokaryotes found in a range of environments. They are infamous for the production of toxins, as well as bioactive compounds, which exhibit anticancer, antimicrobial and protease inhibition activities. Cyanobacteria produce a broad range of antifungals belonging to structural classes, such as peptides, polyketides and alkaloids. Here, we tested cyanobacteria from a wide variety of environments for antifungal activity. The potent antifungal macrolide scytophycin was detected in Anabaena sp. HAN21/1, Anabaena cf. cylindrica PH133, Nostoc sp. HAN11/1 and Scytonema sp. HAN3/2. To our knowledge, this is the first description of Anabaena strains that produce scytophycins. We detected antifungal glycolipopeptide hassallidin production in Anabaena spp. BIR JV1 and HAN7/1 and in Nostoc spp. 6sf Calc and CENA 219. These strains were isolated from brackish and freshwater samples collected in Brazil, the Czech Republic and Finland. In addition, three cyanobacterial strains, Fischerella sp. CENA 298, Scytonema hofmanni PCC 7110 and Nostoc sp. N107.3, produced unidentified antifungal compounds that warrant further characterization. Interestingly, all of the strains shown to produce antifungal compounds in this study belong to Nostocales or Stigonematales cyanobacterial orders.

  19. Compound Wiretap Channels

    Directory of Open Access Journals (Sweden)

    Kramer Gerhard

    2009-01-01

    Full Text Available Abstract This paper considers the compound wiretap channel, which generalizes Wyner's wiretap model to allow the channels to the (legitimate receiver and to the eavesdropper to take a number of possible states. No matter which states occur, the transmitter guarantees that the receiver decodes its message and that the eavesdropper is kept in full ignorance about the message. The compound wiretap channel can also be viewed as a multicast channel with multiple eavesdroppers, in which the transmitter sends information to all receivers and keeps the information secret from all eavesdroppers. For the discrete memoryless channel, lower and upper bounds on the secrecy capacity are derived. The secrecy capacity is established for the degraded channel and the semideterministic channel with one receiver. The parallel Gaussian channel is further studied. The secrecy capacity and the secrecy degree of freedom ( are derived for the degraded case with one receiver. Schemes to achieve the for the case with two receivers and two eavesdroppers are constructed to demonstrate the necessity of a prefix channel in encoder design. Finally, the multi-antenna (i.e., MIMO compound wiretap channel is studied. The secrecy capacity is established for the degraded case and an achievable is given for the general case.

  20. Specificity of specific language impairment

    NARCIS (Netherlands)

    GoorhuisBrouwer, SM; WijnbergWilliams, BJ

    1996-01-01

    In children with specific language impairment (SLI) their problems are supposed to be specifically restricted to language. However, both on a theoretical basis as well as on a practical basis it is often difficult to make a sharp distinction between specific and nonspecific language disorders. In a

  1. Determining the Degree of Promiscuity of Extensively Assayed Compounds

    OpenAIRE

    Jasial, Swarit; Hu, Ye; Bajorath, J?rgen

    2016-01-01

    In the context of polypharmacology, an emerging concept in drug discovery, promiscuity is rationalized as the ability of compounds to specifically interact with multiple targets. Promiscuity of drugs and bioactive compounds has thus far been analyzed computationally on the basis of activity annotations, without taking assay frequencies or inactivity records into account. Most recent estimates have indicated that bioactive compounds interact on average with only one to two targets, whereas dru...

  2. How specific are specific phobias?

    Science.gov (United States)

    Hofmann, S G; Lehman, C L; Barlow, D H

    1997-09-01

    To study the generality of fears among specific phobic individuals and controls, 31 individuals with a DSM-IV diagnosis of specific phobia (natural environmental type: n = 13; blood-injection-injury type: n = 10; and situational type: n = 8) and 33 never mentally ill control subjects participated in an interview and questionnaire study. Based on subjects' fear ratings on the Fear Survey Schedule, subjects were classified as either positive or negative with regard to fear categories that correspond to the five diagnostic subtypes of specific phobia. Phobics showed overall a more generalized form of fear than controls. Furthermore, situational fears were more common among specific phobics who did not meet criteria for specific phobia, situational type, than among controls. These results add to the literature on the functional relationship among different fears and suggest that specific phobias are not as "specific" as is implied by the current diagnostic system.

  3. Offset Compound Gear Drive

    Science.gov (United States)

    Stevens, Mark A.; Handschuh, Robert F.; Lewicki, David G.

    2010-01-01

    The Offset Compound Gear Drive is an in-line, discrete, two-speed device utilizing a special offset compound gear that has both an internal tooth configuration on the input end and external tooth configuration on the output end, thus allowing it to mesh in series, simultaneously, with both a smaller external tooth input gear and a larger internal tooth output gear. This unique geometry and offset axis permits the compound gear to mesh with the smaller diameter input gear and the larger diameter output gear, both of which are on the same central, or primary, centerline. This configuration results in a compact in-line reduction gear set consisting of fewer gears and bearings than a conventional planetary gear train. Switching between the two output ratios is accomplished through a main control clutch and sprag. Power flow to the above is transmitted through concentric power paths. Low-speed operation is accomplished in two meshes. For the purpose of illustrating the low-speed output operation, the following example pitch diameters are given. A 5.0 pitch diameter (PD) input gear to 7.50 PD (internal tooth) intermediate gear (0.667 reduction mesh), and a 7.50 PD (external tooth) intermediate gear to a 10.00 PD output gear (0.750 reduction mesh). Note that it is not required that the intermediate gears on the offset axis be of the same diameter. For this example, the resultant low-speed ratio is 2:1 (output speed = 0.500; product of stage one 0.667 reduction and stage two 0.750 stage reduction). The design is not restricted to the example pitch diameters, or output ratio. From the output gear, power is transmitted through a hollow drive shaft, which, in turn, drives a sprag during which time the main clutch is disengaged.

  4. Boronated porphyrin compounds

    Science.gov (United States)

    Kahl, Stephen B.; Koo, Myoung-Seo

    1992-01-01

    A compound is described having the structure ##STR1## where R preferably is ##STR2## and most preferably R.sup.3 is a closo-carborane and R.sup.2 is --H, an alkyl or aryl having 1 to about 7 carbon atoms, This invention was made with Government support under NIH Grant No. CA-37961 awarded by the Department of Health and Human Services and under the Associated Universities Inc. Contract No. De-AC02-76CH00016 with the U.S. Department of Energy. The Government has rights in this invention.

  5. Frequency effects in compound production

    NARCIS (Netherlands)

    Bien, H.; Levelt, W.J.M.; Baayen, R.H.

    2005-01-01

    Four experiments investigated the role of frequency information in compound production by independently varying the frequencies of the first and second constituent as well as the frequency of the compound itself. Pairs of Dutch noun-noun compounds were selected such that there was a maximal contrast

  6. Organometallic compounds in the environment

    National Research Council Canada - National Science Library

    Craig, P. J

    2003-01-01

    ... of Organometallic Species in the Environment 20 1.10 Stability of Organometallic Compounds in Biological Systems 1.11 G eneral Comments on the Toxicities of Organometallic Compounds 22 1.12 General Considerations on Environmental R eactivity of Organometallic Compounds 24 1.13 Microbial Biotransformation of Metals and M etalloids 25 1.13.1 Introduction 25 1...

  7. Molecular modeling of inorganic compounds

    National Research Council Canada - National Science Library

    Comba, Peter; Hambley, Trevor W; Martin, Bodo

    2009-01-01

    ... mechanics to inorganic and coordination compounds. Initially, simple metal complexes were modeled, but recently the field has been extended to include organometallic compounds, catalysis and the interaction of metal ions with biological macromolecules. The application of molecular mechanics to coordination compounds is complicated by the numbe...

  8. Silane-modified surfaces in specific antibody-mediated cell recognition.

    Science.gov (United States)

    Sterzynska, Karolina; Budna, Joanna; Frydrych-Tomczak, Emilia; Hreczycho, Grzegorz; Malinska, Agnieszka; Maciejewski, Hieronim; Zabel, Maciej

    2014-01-01

    The immobilization of antibodies on various surfaces has been the subject of advanced research in various immunoassay-based diagnostic devices. The physical and chemical stabilities of the immobilized antibodies on a solid surface still cause many problems because upon immobilizing antibody molecules, the antigen-binding ability usually decreases. The silanization of surfaces with organosilanes carrying chemically active groups such as (3-aminopropyl) triethoxysilane (APTES) can accommodate these antigen-binding molecules in an appropriate orientation so that their functionality and binding activity are essentially retained. In this study, n-butyltrimethoxysilane (BMS) and 3-(octafluoropentyloxy)-propyltriethoxysilane (OFPOS) were used as "blocking silanes". The aims of this study were to compare the effectiveness of specific antibody binding of APTES, APTES + BMS and APTES + OFPOS and to characterize the modified surfaces by contact angle measurements and immunofluorescence measurements prior to and after immobilizing proteins. Additionally, we have evaluated the functionality of the immobilized antibodies by their abilities to bind EpCAM-positive human colon adenocarcinoma cell line (LoVo) and EpCAM-negative mouse embryonic fibroblast cell line (3T3). Cell enumeration was conducted on the basis of DAPI-positive signals and recorded using a confocal laser scanning biological microscope. The results of our study showed that the immobilization capability and reactivity of APTES, APTES + BMS and APTES + OFPOS differ. The modification of APTES with unreactive silanes (BMS,OFPOS) is recommended to improve the antibody binding efficiency. However, using OFPOS resulted in more effective antibody and cell binding, and it appears to be the most useful compound in specific antibody-mediated cell recognition.

  9. Specific detection of Lawsonia intracellularis in porcine proliferative enteropathy inferred from fluorescent rRNA in situ hybridization

    DEFF Research Database (Denmark)

    Boye, Mette; Jensen, Tim Kåre; Møller, Kristian

    1998-01-01

    Fluorescent in situ hybridization targeting 16S ribosomal RNA was used for specific detection of the obligate intracellular bacterium Lawsonia intracellularis in enterocytes from pigs affected by proliferative enteropathy. A specific oligonucleotide probe was designed and the specificity...... of the probe was determined by simultaneous comparison with indirect immunofluorescence assay for detection of L. intracellularis in formalin-fixed tissue samples from 15 pigs affected by porcine proliferative enteropathy. We used 10 tissue samples from pigs without proliferative mucosal changes as negative...

  10. Semiconducting III-V compounds

    CERN Document Server

    Hilsum, C; Henisch, Heinz R

    1961-01-01

    Semiconducting III-V Compounds deals with the properties of III-V compounds as a family of semiconducting crystals and relates these compounds to the monatomic semiconductors silicon and germanium. Emphasis is placed on physical processes that are peculiar to III-V compounds, particularly those that combine boron, aluminum, gallium, and indium with phosphorus, arsenic, and antimony (for example, indium antimonide, indium arsenide, gallium antimonide, and gallium arsenide).Comprised of eight chapters, this book begins with an assessment of the crystal structure and binding of III-V compounds, f

  11. Mini-review: Molecular mechanisms of antifouling compounds

    KAUST Repository

    Qian, Pei-Yuan

    2013-04-01

    Various antifouling (AF) coatings have been developed to protect submerged surfaces by deterring the settlement of the colonizing stages of fouling organisms. A review of the literature shows that effective AF compounds with specific targets are ones often considered non-toxic. Such compounds act variously on ion channels, quorum sensing systems, neurotransmitters, production/release of adhesive, and specific enzymes that regulate energy production or primary metabolism. In contrast, AF compounds with general targets may or may not act through toxic mechanisms. These compounds affect a variety of biological activities including algal photosynthesis, energy production, stress responses, genotoxic damage, immunosuppressed protein expression, oxidation, neurotransmission, surface chemistry, the formation of biofilms, and adhesive production/release. Among all the targets, adhesive production/release is the most common, possibly due to a more extensive research effort in this area. Overall, the specific molecular targets and the molecular mechanisms of most AF compounds have not been identified. Thus, the information available is insufficient to draw firm conclusions about the types of molecular targets to be used as sensitive biomarkers for future design and screening of compounds with AF potential. In this review, the relevant advantages and disadvantages of the molecular tools available for studying the molecular targets of AF compounds are highlighted briefly and the molecular mechanisms of the AF compounds, which are largely a source of speculation in the literature, are discussed. © 2013 Copyright Taylor and Francis Group, LLC.

  12. Flavour Compounds in Fungi

    DEFF Research Database (Denmark)

    Ravasio, Davide Antonio

    from the catabolism of amino acids. These compounds are produced by the Ehrlich pathway. The conversion of amino acids into aroma alcohols is accomplished by three enzymatic steps: i) a transamination, ii) a decarboxylation and iii) a dehydration reaction. The transaminase and decarboxylase enzymes...... and 2-phenylacetate. The last part of this thesis presents the initial characterization of twenty non-conventional yeasts (NCY) and their potential application in fermentative processes. These strains have been selected as they have been previously isolated from various fermented food sources....... This selection of strains was used in fermentations with the aim of identifying new interesting flavour producers. Fermentation profiles, volatile analyses, off-flavour identification and resistance to osmotic/oxidative stress have been addressed to highlight new candidates to use for industrial applications...

  13. Flavour Compounds in Fungi

    DEFF Research Database (Denmark)

    Ravasio, Davide Antonio

    provides a tool for the prediction of volatile production in other Saccharomyces sensu stricto species. It can be used to screen a large number of strains for their flavour production within a short time and with low costs and effort. In Ashbya single mutations in the ARO genes led to a strong reduction...... region is directly regulated by the ScAro80 transcription factor. This interaction has been used to create a lacZ-reporter system to correlate the formation of two volatile compounds, 2- phenylethanol and 2-phenylethyl acetate in yeast with ARO9 expression levels. This indirect genetic assay also...... in volatile production, especially in the amount of isoamyl alcohol and 2-phenylethanol. In contrast, the overexpression of the transcriptional regulator AgARO80 did only increase the level of isoamyl alcohol but did not enhance the 2-phenylethanol yield. Promoter analyses of the ARO genes in A. gossypii...

  14. Specifying Specification.

    Science.gov (United States)

    Paulo, Norbert

    2016-03-01

    This paper tackles the accusation that applied ethics is no serious academic enterprise because it lacks theoretical bracing. It does so in two steps. In the first step I introduce and discuss a highly acclaimed method to guarantee stability in ethical theories: Henry Richardson's specification. The discussion shows how seriously ethicists take the stability of the connection between the foundational parts of their theories and their further development as well as their "application" to particular problems or cases. A detailed scrutiny of specification leads to the second step, where I use insights from legal theory to inform the debate around stability from that point of view. This view reveals some of specification's limitations. I suggest that, once specification is sufficiently specified, it appears astonishingly similar to deduction as used in legal theory. Legal theory also provides valuable insight into the functional range of deduction and its relation to other forms of reasoning. This leads to a richer understanding of stability in normative theories and to a smart division of labor between deduction and other forms of reasoning. The comparison to legal theory thereby provides a framework for how different methods such as specification, deduction, balancing, and analogy relate to one another.

  15. Análise de exames de imunofluorescência direta para o diagnóstico de tracoma Analysis of direct immunofluorescence exams for trachoma diagnosis

    Directory of Open Access Journals (Sweden)

    Norma H. Medina

    1996-04-01

    Full Text Available Para confirmação de foco de tracoma em locais onde não existiam casos anteriormente relatados, a Secretaria de Estado da Saúde preconiza a realização de exames laboratoriais principalmente pelo fato da doença ter sido considerada erradicada no Estado de São Paulo, Brasil, na década de 70. Foram colhidas, durante investigações epidemiológicas, lâminas de raspado conjuntival de indivíduos com diagnóstico clínico de tracoma inflamatório (TF/TI. Analisaram-se os resultados dos exames laboratoriais por imunofluorescência direta (IFD, quanto à freqüência de exames adequados para análise e sua positividade segundo a quantidade de corpúsculos elementares (EBs encontrados (sensibilidade do teste. Foram estudadas 385 lâminas, sendo que 241 (62,6% foram consideradas adequadas para análise. Considerando-se o critério de positividade de 5 ou + EBs, a sensibilidade do teste foi de 19,9%. O teste de IFD apesar de ser o melhor teste laboratorial para ser utilizado em trabalhos de campo, não apresenta sensibilidade suficiente para confirmar todos os casos clínicos diagnosticados de tracoma, mas pode confirmar a circulação do agente etiológico em uma comunidade. Portanto, em zonas endêmicas o diagnóstico clínico de tracoma continua sendo o critério para confirmação de casos.For the confirmation of trachoma foci in places where no previous cases had been reported, the State Health Secretariat of S. Paulo makes provision for the realization of laboratory exams, particularly because the disease was considered to have been eradicated from the State in the seventies. During the epidemiological investigations, conjuntival scrapings were collected from the subjects with inflamatory trachoma (TF/TI, clinically diagnosed. The results of the immunofluorescence (DFA exams were analysed in the light of the frequency of the appropriate exams and their positive results, by the quantity of elementary bodies (EB found. A total of 385 slides

  16. Antiviral Screening of Multiple Compounds against Ebola Virus

    Directory of Open Access Journals (Sweden)

    Stuart D. Dowall

    2016-10-01

    Full Text Available In light of the recent outbreak of Ebola virus (EBOV disease in West Africa, there have been renewed efforts to search for effective antiviral countermeasures. A range of compounds currently available with broad antimicrobial activity have been tested for activity against EBOV. Using live EBOV, eighteen candidate compounds were screened for antiviral activity in vitro. The compounds were selected on a rational basis because their mechanisms of action suggested that they had the potential to disrupt EBOV entry, replication or exit from cells or because they had displayed some antiviral activity against EBOV in previous tests. Nine compounds caused no reduction in viral replication despite cells remaining healthy, so they were excluded from further analysis (zidovudine; didanosine; stavudine; abacavir sulphate; entecavir; JB1a; Aimspro; celgosivir; and castanospermine. A second screen of the remaining compounds and the feasibility of appropriateness for in vivo testing removed six further compounds (ouabain; omeprazole; esomeprazole; Gleevec; D-LANA-14; and Tasigna. The three most promising compounds (17-DMAG; BGB324; and NCK-8 were further screened for in vivo activity in the guinea pig model of EBOV disease. Two of the compounds, BGB324 and NCK-8, showed some effect against lethal infection in vivo at the concentrations tested, which warrants further investigation. Further, these data add to the body of knowledge on the antiviral activities of multiple compounds against EBOV and indicate that the scientific community should invest more effort into the development of novel and specific antiviral compounds to treat Ebola virus disease.

  17. Specific Dyslexia.

    Science.gov (United States)

    Naidoo, Sandhya

    The investigation reported in this volume attempts to clarify some issues relating to the existence, nature, and causes of specific dyslexia. Based on an extended study of 98 boys of at least average intelligence with severe reading and spelling problems, the report provides detailed data relating to their developmental and perinatal histories,…

  18. Effect of intermediate compounds and products on wet oxidation and biodegradation rates of pharmaceutical compounds.

    Science.gov (United States)

    Collado, Sergio; Laca, Adriana; Diaz, Mario

    2013-06-01

    Kinetics of pure compounds in batch agitated reactors are useful data to clarify the characteristics of a given reaction, but they frequently do not provide the required information to design industrial mixed continuous processes because in this case the final and intermediate products interact with the reaction of interest, due to backmixing effects. Simultaneously, the presence and transformations of other compounds, frequent in industrial wastewater treatments, adds more complexity to these types of interactions, whose effect can be different, favorable or unfavorable, for chemical or biological reactions. In this work, batch laboratory reactor data were obtained for the wet oxidation and biodegradation of four phenolic compounds present in a pharmaceutical wastewater and then compared with those collected from industrial continuous stirred tank reactors. For wet oxidation, batch laboratory degradation rates were significantly lower than those found in industrial continuous stirred operation. This behavior was explained by a different distribution of intermediate compounds in lab and industrial treatments, caused by the degree of backmixing and the synergistic effects between phenolic compounds (matrix effects). On the other hand, the specific utilization rates during aerobic biodegradation in the continuous industrial operation were lower than those measured in the laboratory, due to the simultaneous presence of the four pollutants in the industrial process (matrix effects) increasing the inhibitory effects of these compounds and its intermediates. Copyright © 2013 Elsevier Ltd. All rights reserved.

  19. Antiviral effect of cationic compounds on bacteriophages

    Directory of Open Access Journals (Sweden)

    Mai Huong eChatain-Ly

    2013-03-01

    Full Text Available The antiviral activity of several cationic compounds - cetytrimethylammonium (CTAB, chitosan, nisin and lysozyme - was investigated on the bacteriophage c2 (DNA head and non-contractile tail infecting Lactococcus strains and the bacteriophage MS2 (F-specific RNA infecting E.coli. Firstly, these activities were evaluated in a phosphate buffer pH 7- 10 mM. The CTAB had a virucidal effect on the Lactococcus bacteriophages, but not on the MS2. After 1 min of contact with 0.125 mM CTAB, the c2 population was reduced from 6 log(pfu/mL to 1,5 log(pfu/mL and completely deactivated at 1 mM. On the contrary, chitosan inhibited the MS2 more than it did the bacteriophages c2. No antiviral effect was observed for the nisin or the lysozyme on bacteriophages after 1 min of treatment. A 1 and 2.5 log reduction was respectively observed for nisin and lysozyme when the treatment time increased (5 or 10 min. These results showed that the antiviral effect depended both on the virus and structure of the antimicrobial compounds. The antiviral activity of these compounds was also evaluated in different physico-chemical conditions and in complex matrices. The antiviral activity of CTAB was impaired in acid pH and with an increase of the ionic strength. These results might be explained by the electrostatic interactions between cationic compounds and negatively charged particles such as bacteriophages or other compounds in a matrix. Milk proved to be protective suggesting the components of food could interfere with antimicrobial compounds.

  20. An immunofluorescence test for diagnosis of ophthalmic herpes in a mouse corneal model Imunofluorescência para diagnóstico de herpes oftálmico usando como modelo córneas de camundongos infectados

    Directory of Open Access Journals (Sweden)

    Sílvia Regina Ferreira Gonçalves Pereira

    2007-04-01

    Full Text Available Herpes simplex virus type 1 (HSV-1 ophthalmic disease is the most common cause of corneal blindness in humans world-wide. Current culture techniques for HSV take several days and commercially available HSV laboratory based diagnostic techniques vary in sensitivity. Our study was conducted to evaluate the use of a quicker and simpler method to herpes ophthalmic diagnosis. Corneal smears were made by firm imprints of infected mouse eyes to glass slides, after smears were fixated with cold acetone, and an indirect immunofluorescence (IIF method was performed using monoclonal antibodies in a murine model of ophthalmic herpes. Eye swabs from infected mice were inoculated in Vero cells for virus isolation. Cytology and histology of the eye were also performed, using hematoxylin-eosin routine. Mouse eyes were examined by slit-lamp biomicroscopy for evidence of herpetic disease at various times postinoculation. We made a comparative evaluation of sensitivity, specificity and speed of methods for laboratory detection of HSV. Our results indicate that this IIF method is quick, sensitive, specific and can be useful in the diagnosis of ophthalmic herpes as demonstrated in an animal model.A doença oftálmica do vírus herpes simplex do tipo 1 (HSV-1 é a causa mais comum de cegueira córnea em humanos mundialmente. Técnicas de cultura atuais para HSV levam vários dias e laboratórios de HSV comercialmente disponíveis estabelecem que as técnicas diagnósticas variam em sensibilidade. Nosso estudo foi conduzido para avaliar a aplicação prática de um método mais rápido e simples para diagnosticar o herpes oftálmico. Decalques córneos foram feitos por impressões firmes de olhos de camundongos a lâminas de vidro, depois os decalques foram fixados com acetona fria, e um método de imunofluorescência indireta (IIF foi executado empregando anticorpos monoclonais no modelo murino de herpes oftálmico. Swabs de córnea foram inoculados em células Vero

  1. Compounding in synthetic aperture imaging

    DEFF Research Database (Denmark)

    Hansen, J. M.; Jensen, J. A.

    2012-01-01

    A method for obtaining compound images using synthetic aperture data is investigated using a convex array transducer. The new approach allows spatial compounding to be performed for any number of angles without reducing the frame rate or temporal resolution. This important feature is an intrinsic...... property of how the compound images are constructed using synthetic aperture data and an improvement compared with how spatial compounding is obtained using conventional methods. The synthetic aperture compound images are created by exploiting the linearity of delay-and-sum beamformation for data collected...... from multiple spherical emissions to synthesize multiple transmit and receive apertures, corresponding to imaging the tissue from multiple directions. The many images are added incoherently, to produce a single compound image. Using a 192-element, 3.5-MHz, λ-pitch transducer, it is demonstrated from...

  2. Optimizing Synthetic Aperture Compound Imaging

    DEFF Research Database (Denmark)

    Hansen, Jens Munk; Jensen, Jørgen Arendt

    2012-01-01

    Spatial compound images are constructed from synthetic aperture data acquired using a linear phased-array transducer. Compound images of wires, tissue, and cysts are created using a method, which allows both transmit and receive compounding without any loss in temporal resolution. Similarly...... to conventional imaging, the speckle reduction achieved by spatial compounding comes at the cost of a reduced detail resolution and a compromise must be made. Using a performance indicator, which can be measured from an image of a phantom without cysts, it is demonstrated how a compromise can be made, which...... is optimal for lesion detection. Synthetic aperture data are acquired from unfocused emissions and 154 compound images are constructed by synthesizing different aperture configurations with more or less compounding, all maintaining a constant resolution across depth corresponding to an f-number of 2...

  3. Pluto's Nonvolatile Chemical Compounds

    Science.gov (United States)

    Grundy, William M.; Binzel, Richard; Cook, Jason C.; Cruikshank, Dale P.; Dalle Ore, Cristina M.; Earle, Alissa M.; Ennico, Kimberly; Jennings, Donald; Howett, Carly; Kaiser, Ralf-Ingo; Linscott, Ivan; Lunsford, A. W.; Olkin, Catherine B.; Parker, Alex Harrison; Parker, Joel Wm.; Philippe, Sylvain; Protopapa, Silvia; Quirico, Eric; Reuter, D. C.; Schmitt, Bernard; Singer, Kelsi N.; Spencer, John R.; Stansberry, John A.; Stern, S. Alan; Tsang, Constantine; Verbiscer, Anne J.; Weaver, Harold A.; Weigle, G. E.; Young, Leslie

    2016-10-01

    Despite the migration of Pluto's volatile ices (N2, CO, and CH4) around the surface on seasonal timescales, the planet's non-volatile materials are not completely hidden from view. They occur in a variety of provinces formed over a wide range of timescales, including rugged mountains and chasms, the floors of mid-latitude craters, and an equatorial belt of especially dark and reddish material typified by the informally named Cthulhu Regio. NASA's New Horizons probe observed several of these regions at spatial resolutions as fine as 3 km/pixel with its LEISA imaging spectrometer, covering wavelengths from 1.25 to 2.5 microns. Various compounds that are much lighter than the tholin-like macromolecules responsible for the reddish coloration, but that are not volatile at Pluto surface temperatures such as methanol (CH3OH) and ethane (C2H6) have characteristic absorption bands within LEISA's wavelength range. This presentation will describe their geographic distributions and attempt to constrain their origins. Possibilities include an inheritance from Pluto's primordial composition (the likely source of H2O ice seen on Pluto's surface) or ongoing production from volatile precursors through photochemistry in Pluto's atmosphere or through radiolysis on Pluto's surface. New laboratory data inform the analysis.This work was supported by NASA's New Horizons project.

  4. Identifiability of Compound Poisson Distributions

    OpenAIRE

    Xekalaki, Evdokia; Panaretos, John

    1983-01-01

    Compound Poisson distributions (CPD's) are frequently used as alternatives in studying situations where a simple Poisson model is found inadequate to describe. In this paper an attempt is made to identify compound Poisson distributions when it is known that the conditional distribution of two random variables (r.v.'s) is compound binomial. Some interesting special cases and their application to accident theory are discussed.

  5. Phenolic compounds in Ecuadorian fruits

    OpenAIRE

    Vasco, Catalina

    2009-01-01

    A group of eighteen fruits cultivated in Ecuador were evaluated for their total soluble phenolic compounds and antioxidant activity and attempts were made to identify the group and content of phenolic compounds responsible for the antioxidant activity. In terms of total phenolic content, three groups (with 1000 mg gallic acid equivalents/100 g FW) were clearly distinguishable. RP-HPLC-DAD and/or LC-MS/MS were used to study the phenolic compounds in four Rosaceae fruits (Andean blackberry, str...

  6. Activity artifacts in drug discovery and different facets of compound promiscuity

    OpenAIRE

    Jürgen Bajorath

    2014-01-01

    Compounds with apparent activity in a variety of assays might disable target proteins or produce false assay signals in the absence of specific interactions. In some instances, such effects are easy to detect, in others they are not. Observed promiscuity of compounds might be due to such non-specific assay artifacts. By contrast, promiscuity might also result from specific interactions with multiple targets. In the latter case, promiscuous compounds can be attractive candidates for certain th...

  7. Natural compounds with herbicidal activity

    Directory of Open Access Journals (Sweden)

    Pasquale Montemurro

    2011-02-01

    Full Text Available Research about phytotoxic activity of natural compounds could lead both to