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06 mai 19221922/05/06 (A51).Appartient à l’ensemble documentaire : PoitouCh

Quantification of KAP transcripts for KAP4, aKAP23 and stKAPy by RT-qPCR.

<p>The relative amount of KAPs was estimated by comparison with the GAPDH expression of WT trypanosomatids. (A) <i>A</i>. <i>deanei</i> KAPs, (B) <i>S</i>. <i>culicis</i> KAPs. APO, aposymbiotic strain; WT, wild type strain. AdKAP4—p > 0.05, aKAP23—p < 0.05 (*), <u>stKAPy</u>—p > 0.05, <u>ScKAP4</u>—p < 0.01 (**).</p

Kinetoplast ultrastructure of WT and APO strains of <i>A</i>. <i>deanei</i> and <i>S</i>. <i>culicis</i>.

<p>A-D: Kinetoplast ultrastructure observed by transmission electron microscopy (TEM). A-B: Both strains of <i>A</i>. <i>deanei</i> present a similar kinetoplast ultrastructure that presents a trapezoid shape and a looser arrangement of the central area in relation to the region that faces the basal body (bb) where kDNA is more condensed (black arrows). C-D: In WT and APO strains of <i>S</i>. <i>culicis</i>, the kinetoplast displays an arch shape and the more condensed layer close to the basal body (black arrows). E-H: Kinetoplast ultrastructure observed by electron tomography (ET). E-F: This high-resolution technique confirmed that both <i>A</i>. <i>deanei</i> strains have a similar kDNA network topology presenting the densely packed kDNA region facing the basal body (brackets). (E, F) top, (E’, F’) middle and (E”, F”) bottom regions of the kinetoplast as visualized by electron tomography. G-H: In both strains of <i>S</i>. <i>culicis</i>, the kinetoplast is flatter when compared to that found in <i>A</i>. <i>deanei</i>, and the compact DNA fibers occupy approximately half of the kDNA network (dashed rectangle). (G, H) top, (G’, H’) middle and (G”, H”) bottom regions of the kinetoplast as visualized by electron tomography. In G’ and H’ arrows indicate the condensed kDNA fibers.</p

Phylogenetic relationships among KAPs as determined by Bayesian inference.

<p>Branches are colored according to the putative KAP type identified. Numbers on nodes represent the Bayesian posterior probability for the corresponding clade (clades without numbers are those presenting 100% posterior probability). Black circles next to taxon names indicate sequences used by Cavalcanti et al. 2009 [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0187516#pone.0187516.ref014" target="_blank">14</a>]; red circles indicate sequences used by Motta et al. 2013 [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0187516#pone.0187516.ref030" target="_blank">30</a>].</p

Immunolocalization of KAPs on WT and APO strains of <i>S</i>. <i>culicis</i> and <i>A</i>. <i>deanei</i>.

<p>DAPI was used to label the nucleus and the kinetoplast, and the anti-porin antibody to identify the symbiotic bacterium in WT cells. (A-B): The anti-KAP4 antibody labeled the kinetoplast region that faces the anterior end of <i>A</i>. <i>deanei</i> in WT (A) and APO (B) cells. Note that this labeling in part coincides with DAPI staining. (C-D): For <i>S</i>. <i>culicis</i>, KAP4 labeling is dispersed through the kinetoplast of both strains and overlaps those for DAPI. (E-F): The anti-aKAP23 antibody generated a similar and specific labeling for WT (E) and APO (F) cells of <i>A</i>. <i>deanei</i>. In this case, an overlap with DAPI was not observed indicating that aKAP23 does not co-localize with the kDNA and is probably located at the KFZ. (G-H) Regarding stKAPy, this protein also faces the anterior part of the cell body in both strains, however the labeling is more disperse when compared to that obtained for aKAP23. Arrowheads indicate the kinetoplast. Scale bars equal to 1 μm.</p

AFM analysis of isolated kDNA networks of <i>A</i>. <i>deanei</i> (A-B) and <i>S</i>. <i>culicis</i> (C-D).

<p>Both species present an arrangement composed of circles uniformly distributed throughout a massive network of kDNA molecules. Clusters of DNA forming rosette-like structures (B and D, arrows) were seen along the network and correspond to regions where the DNA molecules crossover themselves.</p

image_1_Vitamin D Receptor TaqI Polymorphism Is Associated With Reduced Follicle Number in Women Utilizing Assisted Reproductive Technologies.tiff

Purpose<p>Calcitriol, or 1,25-hydroxycholecalciferol, is the active form of vitamin D. It binds and activates vitamin D receptor (VDR). Infertility and defective folliculogenesis have been observed in female vdr-knockout mice; however, whether VDR polymorphisms affect human ovarian responses to controlled ovarian stimulation (COS) remains unclear. We hypothesized that VDR polymorphisms are associated with infertility and COS responses. Thus, we evaluated the association between the TaqI, BsmI, and FokI VDR polymorphisms and ovarian responses in women undergoing COS.</p>Methods<p>In this study, we recruited a control group (n = 121) comprising volunteers with a history of natural conception and a second group of women undergoing COS (n = 70). TaqI, BsmI, and FokI genotyping was performed via restriction fragment length polymorphism analysis or TaqMan qPCR and Sanger sequencing. Intrafollicular 25(OH)D contents were measured in follicular fluid collected from COS patients during oocyte retrieval. Ovarian response parameters were obtained from patient medical records.</p>Results<p>There were no significant differences in the genotype frequencies of VDR polymorphisms (TaqI, BsmI and FokI) between the control and COS groups. However, the allele frequency of TaqI (C allele) was significantly lower in the COS group than in the control group (p = 0.02). Follicle number but not oocyte number was lower in patients with TaqI polymorphic (TC/CC) genotypes (p = 0.03). Importantly, the ratio between the number of follicles retrieved and intrafollicular estradiol concentrations was higher in patients with the TC/CC TaqI genotypes (p < 0.02).</p>Conclusion<p>We identified an association between the VDR TaqI polymorphism and reduced follicle number in women undergoing COS, suggesting that VDR signaling affects the ovarian response to stimulation via unknown mechanisms.</p

image_3_Vitamin D Receptor TaqI Polymorphism Is Associated With Reduced Follicle Number in Women Utilizing Assisted Reproductive Technologies.tiff

Purpose<p>Calcitriol, or 1,25-hydroxycholecalciferol, is the active form of vitamin D. It binds and activates vitamin D receptor (VDR). Infertility and defective folliculogenesis have been observed in female vdr-knockout mice; however, whether VDR polymorphisms affect human ovarian responses to controlled ovarian stimulation (COS) remains unclear. We hypothesized that VDR polymorphisms are associated with infertility and COS responses. Thus, we evaluated the association between the TaqI, BsmI, and FokI VDR polymorphisms and ovarian responses in women undergoing COS.</p>Methods<p>In this study, we recruited a control group (n = 121) comprising volunteers with a history of natural conception and a second group of women undergoing COS (n = 70). TaqI, BsmI, and FokI genotyping was performed via restriction fragment length polymorphism analysis or TaqMan qPCR and Sanger sequencing. Intrafollicular 25(OH)D contents were measured in follicular fluid collected from COS patients during oocyte retrieval. Ovarian response parameters were obtained from patient medical records.</p>Results<p>There were no significant differences in the genotype frequencies of VDR polymorphisms (TaqI, BsmI and FokI) between the control and COS groups. However, the allele frequency of TaqI (C allele) was significantly lower in the COS group than in the control group (p = 0.02). Follicle number but not oocyte number was lower in patients with TaqI polymorphic (TC/CC) genotypes (p = 0.03). Importantly, the ratio between the number of follicles retrieved and intrafollicular estradiol concentrations was higher in patients with the TC/CC TaqI genotypes (p < 0.02).</p>Conclusion<p>We identified an association between the VDR TaqI polymorphism and reduced follicle number in women undergoing COS, suggesting that VDR signaling affects the ovarian response to stimulation via unknown mechanisms.</p

General characteristics of the <i>T. rangeli</i> genome.

<p>* Excluding proteins of unknown function.</p><p>General characteristics of the <i>T. rangeli</i> genome.</p

Representation of the telomeric and subtelomeric regions of <i>Trypanosoma rangeli</i>, <i>T. cruzi</i> and <i>T. brucei</i>.

<p>The two types of telomeres identified in <i>T. rangeli</i> and two others representing the heterogeneity of <i>T. cruzi</i> chromosome ends are shown. The size of the subtelomeric region, which extends between the telomeric hexamer repeats and the first internal core genes of the trypanosomes, is indicated below each map. Boxes indicate genes and/or gene arrays. The maps are not to scale. The <i>T. brucei</i> and <i>T. cruzi</i> maps were adapted from <a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0003176#pntd.0003176-ElSayed1" target="_blank">[55]</a>, <a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0003176#pntd.0003176-MoraesBarros1" target="_blank">[98]</a>.</p