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Tiana Tasca - One of the best experts on this subject based on the ideXlab platform.

  • Expression stability values (M) and ranking of the candidate reference genes based on geNorm calculation.
    2015
    Co-Authors: Odelta Dos Santos, Graziela De Vargas Rigo, Amanda Piccoli Frasson, Alexandre José Macedo, Tiana Tasca
    Abstract:

    (a) total samples; (b) samples under all nutrient restriction; (c) samples with 1% HIBS; (d) maltose restriction; (e) HIBS and maltose restriction; (f) samples supplemented with 200 μM Ferrous Ammonium Sulfate. The M value and ranking were calculated through a pairwise comparison and stepwise exclusion of the lowest stable gene. Low M values correspond to high expression stability.

  • The quantification of pyruvate-ferredoxin oxidoreductase (PFOR) gene expression in T. vaginalis TV-LACM6 isolate.
    2015
    Co-Authors: Odelta Dos Santos, Graziela De Vargas Rigo, Amanda Piccoli Frasson, Alexandre José Macedo, Tiana Tasca
    Abstract:

    The relative expression of PFOR gene in T. vaginalis under Ferrous Ammonium Sulfate (high-iron condition 100, 200, and 300 μM) using GAPDH, and DNATopII as internal controls, after 1, 6, 12, and 24h of cultivation. (a) The relative expression of PFOR gene in T. vaginalis under high-iron after 1 hour of cultivation; (b) The relative expression of PFOR gene in T. vaginalis under high-iron condition after 6 hours of cultivation; (c) The relative expression of PFOR gene in T. vaginalis under high-iron condition after 12 hours of cultivation; (d) The relative expression of PFOR gene in T. vaginalis under high-iron condition after 24 hours of cultivation. The relative expression levels are depicted as the mean ± SD, calculated from three biological replicate. The relative change in gene expression was analysed using the 2-ΔΔCt method. Statistically significant expression changes were calculated using one-way ANOVA and the level of significance was also determined by the Bonferroni method comparing all groups versus the control. Statistically significance (P

  • The quantification of pyruvate-ferredoxin oxidoreductase (PFOR) gene expression in T. vaginalis ATCC 30238 isolate.
    2015
    Co-Authors: Odelta Dos Santos, Graziela De Vargas Rigo, Amanda Piccoli Frasson, Alexandre José Macedo, Tiana Tasca
    Abstract:

    The relative expression of PFOR gene in T. vaginalis under Ferrous Ammonium Sulfate (high-iron condition 100, 200, and 300 μM) using GAPDH, and DNATopII as internal controls, after 1, 6, 12, and 24h of cultivation. (a) The relative expression of PFOR gene in T. vaginalis under high-iron after 1 hour of cultivation; (b) The relative expression of PFOR gene in T. vaginalis under high-iron condition after 6 hours of cultivation; (c) The relative expression of PFOR gene in T. vaginalis under high-iron condition after 12 hours of cultivation; (d) The relative expression of PFOR gene in T. vaginalis under high-iron condition after 24 hours of cultivation. The relative expression levels are depicted as the mean ± SD, calculated from three biological replicate. The relative change in gene expression was analysed using the 2-ΔΔCt method. Statistically significant expression changes were calculated using one-way ANOVA and the level of significance was also determined by the Bonferroni method comparing all groups versus the control. Statistically significance (P

  • The quantification of pyruvate-ferredoxin oxidoreductase (PFOR) gene expression in T. vaginalis TV-LACH4 isolate.
    2015
    Co-Authors: Odelta Dos Santos, Graziela De Vargas Rigo, Amanda Piccoli Frasson, Alexandre José Macedo, Tiana Tasca
    Abstract:

    The relative expression of PFOR gene in T. vaginalis under Ferrous Ammonium Sulfate (high-iron condition 100, 200, and 300 μM) using GAPDH, and DNATopII as internal controls, after 1, 6, 12, and 24h of cultivation. (a) The relative expression of PFOR gene in T. vaginalis under high-iron after 1 hour of cultivation; (b) The relative expression of PFOR gene in T. vaginalis under high-iron condition after 6 hours of cultivation; (c) The relative expression of PFOR gene in T. vaginalis under high-iron condition after 12 hours of cultivation; (d) The relative expression of PFOR gene in T. vaginalis under high-iron condition after 24 hours of cultivation. The relative expression levels are depicted as the mean ± SD, calculated from three biological replicate. The relative change in gene expression was analyzed using the 2-ΔΔCt method. Statistically significant expression changes were calculated using one-way ANOVA and the level of significance was also determined by the Bonferroni method comparing all groups versus the control. Statistically significance (P

  • Optimal Reference Genes for Gene Expression Normalization in Trichomonas vaginalis
    2015
    Co-Authors: Odelta Dos Santos, Graziela De Vargas Rigo, Amanda Piccoli Frasson, Alexandre José Macedo, Tiana Tasca
    Abstract:

    Trichomonas vaginalis is the etiologic agent of trichomonosis, the most common non-viral sexually transmitted disease worldwide. This infection is associated with several health consequences, including cervical and prostate cancers and HIV acquisition. Gene expression analysis has been facilitated because of available genome sequences and large-scale transcriptomes in T. vaginalis, particularly using quantitative real-time polymerase chain reaction (qRT-PCR), one of the most used methods for molecular studies. Reference genes for normalization are crucial to ensure the accuracy of this method. However, to the best of our knowledge, a systematic validation of reference genes has not been performed for T. vaginalis. In this study, the transcripts of nine candidate reference genes were quantified using qRT-PCR under different cultivation conditions, and the stability of these genes was compared using the geNorm and NormFinder algorithms. The most stable reference genes were α-tubulin, actin and DNATopII, and, conversely, the widely used T. vaginalis reference genes GAPDH and β-tubulin were less stable. The PFOR gene was used to validate the reliability of the use of these candidate reference genes. As expected, the PFOR gene was upregulated when the trophozoites were cultivated with Ferrous Ammonium Sulfate when the DNATopII, α-tubulin and actin genes were used as normalizing gene. By contrast, the PFOR gene was downregulated when the GAPDH gene was used as an internal control, leading to misinterpretation of the data. These results provide an important starting point for reference gene selection and gene expression analysis with qRT-PCR studies of T. vaginalis.

Odelta Dos Santos - One of the best experts on this subject based on the ideXlab platform.

  • RESEARCH ARTICLE Optimal Reference Genes for Gene Expression Normalization in Trichomonas vaginalis
    2016
    Co-Authors: Odelta Dos Santos, Graziela De Vargas Rigo, Piccoli A Frasson
    Abstract:

    Trichomonas vaginalis is the etiologic agent of trichomonosis, the most common non-viral sexually transmitted disease worldwide. This infection is associated with several health con-sequences, including cervical and prostate cancers and HIV acquisition. Gene expression analysis has been facilitated because of available genome sequences and large-scale tran-scriptomes in T. vaginalis, particularly using quantitative real-time polymerase chain reaction (qRT-PCR), one of the most used methods for molecular studies. Reference genes for nor-malization are crucial to ensure the accuracy of this method. However, to the best of our knowledge, a systematic validation of reference genes has not been performed for T. vagina-lis. In this study, the transcripts of nine candidate reference genes were quantified using qRT-PCR under different cultivation conditions, and the stability of these genes was com-pared using the geNorm and NormFinder algorithms. The most stable reference genes were α-tubulin, actin andDNATopII, and, conversely, the widely used T. vaginalis reference genesGAPDH and β-tubulin were less stable. The PFOR gene was used to validate the reli-ability of the use of these candidate reference genes. As expected, the PFOR gene was upregulated when the trophozoites were cultivated with Ferrous Ammonium Sulfate when the DNATopII, α-tubulin and actin genes were used as normalizing gene. By contrast, the PFOR gene was downregulated when theGAPDH gene was used as an internal control, leading to misinterpretation of the data. These results provide an important starting point for reference gene selection and gene expression analysis with qRT-PCR studies of T. vaginalis

  • Expression stability values (M) and ranking of the candidate reference genes based on geNorm calculation.
    2015
    Co-Authors: Odelta Dos Santos, Graziela De Vargas Rigo, Amanda Piccoli Frasson, Alexandre José Macedo, Tiana Tasca
    Abstract:

    (a) total samples; (b) samples under all nutrient restriction; (c) samples with 1% HIBS; (d) maltose restriction; (e) HIBS and maltose restriction; (f) samples supplemented with 200 μM Ferrous Ammonium Sulfate. The M value and ranking were calculated through a pairwise comparison and stepwise exclusion of the lowest stable gene. Low M values correspond to high expression stability.

  • The quantification of pyruvate-ferredoxin oxidoreductase (PFOR) gene expression in T. vaginalis TV-LACM6 isolate.
    2015
    Co-Authors: Odelta Dos Santos, Graziela De Vargas Rigo, Amanda Piccoli Frasson, Alexandre José Macedo, Tiana Tasca
    Abstract:

    The relative expression of PFOR gene in T. vaginalis under Ferrous Ammonium Sulfate (high-iron condition 100, 200, and 300 μM) using GAPDH, and DNATopII as internal controls, after 1, 6, 12, and 24h of cultivation. (a) The relative expression of PFOR gene in T. vaginalis under high-iron after 1 hour of cultivation; (b) The relative expression of PFOR gene in T. vaginalis under high-iron condition after 6 hours of cultivation; (c) The relative expression of PFOR gene in T. vaginalis under high-iron condition after 12 hours of cultivation; (d) The relative expression of PFOR gene in T. vaginalis under high-iron condition after 24 hours of cultivation. The relative expression levels are depicted as the mean ± SD, calculated from three biological replicate. The relative change in gene expression was analysed using the 2-ΔΔCt method. Statistically significant expression changes were calculated using one-way ANOVA and the level of significance was also determined by the Bonferroni method comparing all groups versus the control. Statistically significance (P

  • The quantification of pyruvate-ferredoxin oxidoreductase (PFOR) gene expression in T. vaginalis ATCC 30238 isolate.
    2015
    Co-Authors: Odelta Dos Santos, Graziela De Vargas Rigo, Amanda Piccoli Frasson, Alexandre José Macedo, Tiana Tasca
    Abstract:

    The relative expression of PFOR gene in T. vaginalis under Ferrous Ammonium Sulfate (high-iron condition 100, 200, and 300 μM) using GAPDH, and DNATopII as internal controls, after 1, 6, 12, and 24h of cultivation. (a) The relative expression of PFOR gene in T. vaginalis under high-iron after 1 hour of cultivation; (b) The relative expression of PFOR gene in T. vaginalis under high-iron condition after 6 hours of cultivation; (c) The relative expression of PFOR gene in T. vaginalis under high-iron condition after 12 hours of cultivation; (d) The relative expression of PFOR gene in T. vaginalis under high-iron condition after 24 hours of cultivation. The relative expression levels are depicted as the mean ± SD, calculated from three biological replicate. The relative change in gene expression was analysed using the 2-ΔΔCt method. Statistically significant expression changes were calculated using one-way ANOVA and the level of significance was also determined by the Bonferroni method comparing all groups versus the control. Statistically significance (P

  • The quantification of pyruvate-ferredoxin oxidoreductase (PFOR) gene expression in T. vaginalis TV-LACH4 isolate.
    2015
    Co-Authors: Odelta Dos Santos, Graziela De Vargas Rigo, Amanda Piccoli Frasson, Alexandre José Macedo, Tiana Tasca
    Abstract:

    The relative expression of PFOR gene in T. vaginalis under Ferrous Ammonium Sulfate (high-iron condition 100, 200, and 300 μM) using GAPDH, and DNATopII as internal controls, after 1, 6, 12, and 24h of cultivation. (a) The relative expression of PFOR gene in T. vaginalis under high-iron after 1 hour of cultivation; (b) The relative expression of PFOR gene in T. vaginalis under high-iron condition after 6 hours of cultivation; (c) The relative expression of PFOR gene in T. vaginalis under high-iron condition after 12 hours of cultivation; (d) The relative expression of PFOR gene in T. vaginalis under high-iron condition after 24 hours of cultivation. The relative expression levels are depicted as the mean ± SD, calculated from three biological replicate. The relative change in gene expression was analyzed using the 2-ΔΔCt method. Statistically significant expression changes were calculated using one-way ANOVA and the level of significance was also determined by the Bonferroni method comparing all groups versus the control. Statistically significance (P

Kevin Jordan - One of the best experts on this subject based on the ideXlab platform.

  • an apparent threshold dose response in Ferrous xylenol orange gel dosimeters when scanned with a yellow light source
    2008
    Co-Authors: S Babic, J J Battista, Kevin Jordan
    Abstract:

    Freshly prepared radiochromic Ferrous xylenol-orange (FX) gels optically scanned with a light source exhibit a threshold dose response that is thermally and wavelength dependent. Correction for this threshold dose leads to accurate dose calibration and better reproducibility in multiple fraction radiation exposures. The objective of this study was to determine the cause of the threshold dose effect and to control it through improved dose calibration procedures. The results of a systematic investigation into the chemical cause revealed that impurities within the various FX gel constituents (i.e. xylenol-orange, gelatin, sulfuric acid and Ferrous Ammonium Sulfate) were not directly responsible for the threshold dose. Rather, it was determined that the threshold dose response stems from a spectral sensitivity to different chemical complexes that are formed at different dose levels in FX gels between ferric (Fe(III)) ions and xylenol-orange (XO), i.e. Fe(III)i:XOj. A double Fe(III)2:XO1 complex preferentially absorbs at longer wavelengths (i.e. yellow), while at shorter wavelengths (i.e. green) the sensitivity is biased toward the single Fe(III)1:XO1 complex. As a result, when scanning with yellow light, freshly prepared FX gels require a minimum concentration of Fe(III) ions to shift the equilibrium concentration to favor the predominant production of the double Fe(III)2:XO1 complex at low doses. This can be accomplished via pre-irradiation of freshly prepared gels to a priming dose of ~0.5 Gy or allowing auto-oxidation to generate the startup concentration of Fe(III) ions required to negate the apparent threshold dose response.

A Negronmendoza - One of the best experts on this subject based on the ideXlab platform.

  • study of the dose response of the system Ferrous Ammonium Sulfate sucrose xylenol orange in acid aqueous solution
    2014
    Co-Authors: J M Juarezcalderon, A Negronmendoza, S Ramosbernal
    Abstract:

    Abstract An aqueous solution of Ammonium Ferrous Sulfate–sucrose–xylenol orange in sulfuric acid (FSX) is proposed as a dosimetric system for the processes of gamma irradiation in a range between 0.3 and 6 Gy. This system is based on the indirect oxidation of Ferrous ion by an organic compound (sucrose) to ferric ion and on the formation of a color complex of Fe 3+ in an acidic medium with xylenol orange (a dye). After gamma radiation, an observable change occurs in the color of the system. Irradiation was executed at three different temperatures (13 °C, 22 °C, and 40 °C). A spectrometric readout method at 585 nm was employed to evaluate the system׳s dose response. In all of the cases analyzed, the responses had a linear behavior, and a slight effect of irradiation temperature was observed. Post-irradiation response was also evaluated and showed the stability of the solutions 24 h after the irradiation. The results obtained suggest that FSX might be used as a dosimeter for low doses of gamma irradiation because it provides a stable signal, good reproducibility, and an accessible technique for analysis.

  • evaluation of commercial grade Ferrous Ammonium Sulfate as potential dosimeter for technological irradiations
    2008
    Co-Authors: Manuel J Juarezcalderon, S Ramosbernal, A Negronmendoza
    Abstract:

    In this work, we have studied the behavior of crystalline Ferrous Ammonium Sulfate (FAS) under gamma irradiation. The doses studied ranged from 33.5 to 270 kGy. The purpose of this study is to explain the setup, measurement, and reporting procedures for using FAS as a dosimeter.The results obtained in the present study show that this salt very easily gave reproducible results, a linear response, as well as, simple sample preparation and reading. The irradiation temperature and dose rate in the response of the iron salt were found to have slight influence. Any storage time up to 18 months resulted in a 36% decrease in the response. The variation in the response obtained for short periods of storage is negligible for this application. Due to the properties of the FAS system also can be use as transfer dosismeter.

  • irradiation of Ferrous Ammonium Sulfate for its use as high absorbed dose and low temperature dosimeter
    2007
    Co-Authors: J M Juarezcalderon, A Negronmendoza, S Ramosbernal
    Abstract:

    Abstract In the present paper, we study the response of crystalline Ammonium Ferrous Sulfate as a function of the irradiation dose and temperature. The dose studied ranged from 33.5 to 546 kGy. The temperature regimen varied from 77 K (liquid nitrogen) to 311 K. The analysis of the samples was made by UV spectroscopy and EPR. The results show that the change in absorbance of the dosimeter was linear with respect to the absorbed dose in the range studied. There is a small influence of the irradiation temperature in the response of the iron salt. The dose rate and storage time after irradiation was of no importance in this application.

Graziela De Vargas Rigo - One of the best experts on this subject based on the ideXlab platform.

  • RESEARCH ARTICLE Optimal Reference Genes for Gene Expression Normalization in Trichomonas vaginalis
    2016
    Co-Authors: Odelta Dos Santos, Graziela De Vargas Rigo, Piccoli A Frasson
    Abstract:

    Trichomonas vaginalis is the etiologic agent of trichomonosis, the most common non-viral sexually transmitted disease worldwide. This infection is associated with several health con-sequences, including cervical and prostate cancers and HIV acquisition. Gene expression analysis has been facilitated because of available genome sequences and large-scale tran-scriptomes in T. vaginalis, particularly using quantitative real-time polymerase chain reaction (qRT-PCR), one of the most used methods for molecular studies. Reference genes for nor-malization are crucial to ensure the accuracy of this method. However, to the best of our knowledge, a systematic validation of reference genes has not been performed for T. vagina-lis. In this study, the transcripts of nine candidate reference genes were quantified using qRT-PCR under different cultivation conditions, and the stability of these genes was com-pared using the geNorm and NormFinder algorithms. The most stable reference genes were α-tubulin, actin andDNATopII, and, conversely, the widely used T. vaginalis reference genesGAPDH and β-tubulin were less stable. The PFOR gene was used to validate the reli-ability of the use of these candidate reference genes. As expected, the PFOR gene was upregulated when the trophozoites were cultivated with Ferrous Ammonium Sulfate when the DNATopII, α-tubulin and actin genes were used as normalizing gene. By contrast, the PFOR gene was downregulated when theGAPDH gene was used as an internal control, leading to misinterpretation of the data. These results provide an important starting point for reference gene selection and gene expression analysis with qRT-PCR studies of T. vaginalis

  • Expression stability values (M) and ranking of the candidate reference genes based on geNorm calculation.
    2015
    Co-Authors: Odelta Dos Santos, Graziela De Vargas Rigo, Amanda Piccoli Frasson, Alexandre José Macedo, Tiana Tasca
    Abstract:

    (a) total samples; (b) samples under all nutrient restriction; (c) samples with 1% HIBS; (d) maltose restriction; (e) HIBS and maltose restriction; (f) samples supplemented with 200 μM Ferrous Ammonium Sulfate. The M value and ranking were calculated through a pairwise comparison and stepwise exclusion of the lowest stable gene. Low M values correspond to high expression stability.

  • The quantification of pyruvate-ferredoxin oxidoreductase (PFOR) gene expression in T. vaginalis TV-LACM6 isolate.
    2015
    Co-Authors: Odelta Dos Santos, Graziela De Vargas Rigo, Amanda Piccoli Frasson, Alexandre José Macedo, Tiana Tasca
    Abstract:

    The relative expression of PFOR gene in T. vaginalis under Ferrous Ammonium Sulfate (high-iron condition 100, 200, and 300 μM) using GAPDH, and DNATopII as internal controls, after 1, 6, 12, and 24h of cultivation. (a) The relative expression of PFOR gene in T. vaginalis under high-iron after 1 hour of cultivation; (b) The relative expression of PFOR gene in T. vaginalis under high-iron condition after 6 hours of cultivation; (c) The relative expression of PFOR gene in T. vaginalis under high-iron condition after 12 hours of cultivation; (d) The relative expression of PFOR gene in T. vaginalis under high-iron condition after 24 hours of cultivation. The relative expression levels are depicted as the mean ± SD, calculated from three biological replicate. The relative change in gene expression was analysed using the 2-ΔΔCt method. Statistically significant expression changes were calculated using one-way ANOVA and the level of significance was also determined by the Bonferroni method comparing all groups versus the control. Statistically significance (P

  • The quantification of pyruvate-ferredoxin oxidoreductase (PFOR) gene expression in T. vaginalis ATCC 30238 isolate.
    2015
    Co-Authors: Odelta Dos Santos, Graziela De Vargas Rigo, Amanda Piccoli Frasson, Alexandre José Macedo, Tiana Tasca
    Abstract:

    The relative expression of PFOR gene in T. vaginalis under Ferrous Ammonium Sulfate (high-iron condition 100, 200, and 300 μM) using GAPDH, and DNATopII as internal controls, after 1, 6, 12, and 24h of cultivation. (a) The relative expression of PFOR gene in T. vaginalis under high-iron after 1 hour of cultivation; (b) The relative expression of PFOR gene in T. vaginalis under high-iron condition after 6 hours of cultivation; (c) The relative expression of PFOR gene in T. vaginalis under high-iron condition after 12 hours of cultivation; (d) The relative expression of PFOR gene in T. vaginalis under high-iron condition after 24 hours of cultivation. The relative expression levels are depicted as the mean ± SD, calculated from three biological replicate. The relative change in gene expression was analysed using the 2-ΔΔCt method. Statistically significant expression changes were calculated using one-way ANOVA and the level of significance was also determined by the Bonferroni method comparing all groups versus the control. Statistically significance (P

  • The quantification of pyruvate-ferredoxin oxidoreductase (PFOR) gene expression in T. vaginalis TV-LACH4 isolate.
    2015
    Co-Authors: Odelta Dos Santos, Graziela De Vargas Rigo, Amanda Piccoli Frasson, Alexandre José Macedo, Tiana Tasca
    Abstract:

    The relative expression of PFOR gene in T. vaginalis under Ferrous Ammonium Sulfate (high-iron condition 100, 200, and 300 μM) using GAPDH, and DNATopII as internal controls, after 1, 6, 12, and 24h of cultivation. (a) The relative expression of PFOR gene in T. vaginalis under high-iron after 1 hour of cultivation; (b) The relative expression of PFOR gene in T. vaginalis under high-iron condition after 6 hours of cultivation; (c) The relative expression of PFOR gene in T. vaginalis under high-iron condition after 12 hours of cultivation; (d) The relative expression of PFOR gene in T. vaginalis under high-iron condition after 24 hours of cultivation. The relative expression levels are depicted as the mean ± SD, calculated from three biological replicate. The relative change in gene expression was analyzed using the 2-ΔΔCt method. Statistically significant expression changes were calculated using one-way ANOVA and the level of significance was also determined by the Bonferroni method comparing all groups versus the control. Statistically significance (P