Effect of Oocyte Vitrification Before and After in Vitro Maturation Towards Bcl-2, Bax and Bcl-2/Bax Ratio Expression

Objectives: to compare the expression of Bcl-2, Bax and Bcl-2/Bax ratio in cumulus cell and oocyte between vitrified oocyte pre and post in vitro maturation.Materials and Methods: Maturation was operated in medium TC 100 µl for 24 hours. Vitrification begins with washing oocyte in PBS basic medium supplemented of 20% serum for 1-2 minutes, followed by equilibration medium PBS + 20% serum + 10% ethylene glycol for 10-14 minutes, then transferred to 20% serum + PBS + 0.5 M sucrose + 15% ethylene glycol + PROH 15% for 25-30 seconds. Thawing is processed by submerging the oocytes in the media: 1). PBS + 20% serum + 0.5 M sucrose, 2). PBS + 20% serum + 0.25 M sucrose, and 3). PBS + 20% serum + 0.1 M sucrose. Imunocytochemistry observed the expression of Bcl-2, bax and Bcl-2/bax ratio.Results: Bcl-2 expression on oocyte in control group differed significantly with treatment group, Bcl-2 expression on cumulus in control group differed significantly with treatment 1 group. Bax expression on oocyte in control group differed significantly with treatment group. Bax expression on cumulus in control group differed significantly with treatment group. Bcl-2/Bax expression ratio on oocyte and cumulus did not differ significantly in all groupConclusion: No difference Bcl-2/Bax expression ratio on oocyte and cumulus between vitrified oocyte pre and post in vitro maturation

A Novel Mechanism for CTCF in the Epigenetic Regulation of Bax in Breast Cancer Cells

Wepreviously reported the association of elevated levels of themultifunctional transcription factor, CCCTC binding factor (CTCF), in breast cancer cells with the specific anti-apoptotic function of CTCF. To understand the molecularmechanisms of this phenomenon, we investigated regulation of the human Bax gene by CTCF in breast and non-breast cells. Two CTCF binding sites (CTSs) within the Bax promoter were identified. In all cells, breast and non-breast, active histone modifications were present at these CTSs, DNA harboring this region was unmethylated, and levels of Bax mRNA and protein were similar. Nevertheless, up-regulation of Bax mRNA and protein and apoptotic cell deathwere observed only in breast cancer cells depleted of CTCF.We proposed that increased CTCF binding to the Bax promoter in breast cancer cells, by comparison with non-breast cells, may be mechanistically linked to the specific apoptotic phenotype in CTCF-depleted breast cancer cells. In this study, we show that CTCF binding was enriched at the Bax CTSs in breast cancer cells and tumors; in contrast, binding of other transcription factors (SP1,WT1, EGR1, and c-Myc) was generally increased in non- breast cells and normal breast tissues. Our findings suggest a novel mechanism for CTCF in the epigenetic regulation of Bax in breast cancer cells, whereby elevated levels of CTCF support preferential binding of CTCF to the Bax CTSs. In this context, CTCF functions as a transcriptional repressor counteracting influences of positive regulatory factors; depletion of breast cancer cells from CTCF therefore results in the activation of Bax and apoptosis. © 2013 Neoplasia Press, Inc

Distinct promoter elements mediate the co-operative effect of Brn-3a and p53 on the p21 promoter and their antagonism on the Bax promoter

Although the promoters of both the Bax and p21 genes are activated by p53, they differ in the effect on this activation of the POU family transcription factor Brn-3a. Thus, Brn-3a inhibits activation of the Bax promoter by p53 but enhances the ability of p53 to activate the p21 promoter. We demonstrate that repression of p53-mediated activation of the Bax promoter involves a complex upstream sequence in which two Brn-3a response elements flank the p53 response element. In contrast, a minimal p21 promoter is activated by Brn-3a and such activation cannot be abolished without abolishing basal promoter activity. Moreover, synergistic activation by Brn-3a and p53 continues to be observed when the p53-binding sites in the p21 promoter are substituted by the Bax p53 site or by the region of the Bax promoter essential for Brn-3a-mediated repression, indicating that the p21 core promoter plays a central role in this response. The significance of these effects is discussed in terms of the different responses of the Bax and p21 promoters and the overlapping but distinct roles of Brn-3a and p53 in neuronal growth arrest and apoptosis

RelA/NF-kappaB recruitment on the bax gene promoter antagonizes p73-dependent apoptosis in costimulated T cells

The balance between antiapoptotic and proapoptotic proteins of the Bcl-2 family is critical in determining the fate of T cells in response to death stimuli. Proapoptotic genes, such as bax, are generally regulated by the p53 family of transcription factors, whereas NF-kappaB subunits can activate the transcription of antiapoptotic Bcl-2 members. Here, we show that CD28 activation protects memory T cells from irradiation-induced apoptosis by both upregulating bcl-xL and inhibiting bax gene expression. We found that p73, but not p53, binds to and trans-activates the bax gene promoter in irradiated T cells. The activation of RelA/NF-kappaB subunit in CD28 costimulated T cells and its binding onto the bax gene promoter results in suppression of bax transcription and decrease in both p73 and RNA polymerase II recruitment in vivo. RelA recruitment on the bax gene promoter is also accompanied by the lost of p300 binding and the parallel appearance of histone deacetylase-1-containing complexes. These findings identify RelA/NF-kappaB as a critical regulator of T-cell survival by affecting the balance of Bcl-2 family members

Introducing BAX: a database for X-ray clusters and groups of galaxies

We present BAX, Base de Donnees Amas de Galaxies X (http://webast.ast.obs-mip.fr/bax), a multi-wavelength database dedicated to X-ray clusters and groups of galaxies allowing detailed information retrieval. BAX is designed to support astronomical research by providing access to published measurements of the main physical quantities and to the related bibliographic references: basic data stored in the database are cluster/group identifiers, equatorial coordinates, redshift, flux, X-ray luminosity (in the ROSAT band) and temperature, and links to additional linked parameters (in X-rays, such as spatial profile parameters, as well as SZ parameters of the hot gas, lensing measurements,and data at other wavelengths, such as optical and radio). The clusters and groups in BAX can be queried by the basic parameters as well as the linked parameters or combinations of these. We expect BAX to become an important tool for the astronomical community. BAX will optimize various aspects of the scientific analysis of X-ray clusters and groups of galaxies, from proposal planning to data collection, interpretation and publication, from both ground based facilities like MEGACAM (CFHT), VIRMOS (VLT) and space missions like XMM-Newton, Chandra and Planck.Comment: Accepted for publication in Astronomy and Astrophysics Journal. Contains 4 pages and 1 figur

Readers’ cognitive processes during IELTS reading tests: evidence from eye tracking

The research described in this report investigates readers' mental processes as they complete onscreen IELTS (International English Language Testing System) reading test items. It employs up-to-date eye tracking technology to research readers' eye movements and aims, among other things, to contribute to an understanding of the cognitive validity of reading test items (Glaser. 1991; Field forthcoming). Participants were a group of Malaysian undergraduates (n=71) taking an onscreen test consisting of two IELTS reading passages with a total of 11 test items. The eye movements of a random sample of these participants (n=38) were tracked. Questionnaire and stimulated recall interview data were also collected, and were important in order to interpret and explain the eye tracking data. Findings demonstrated significant differences between successful and unsuccessful test-takers on a number of dimensions, including their ability to read expeditiously (Khalifa and Weir. 2009). and their focus on particular aspects of the test items and the reading texts. This demonstrates the potential of eye tracking, in combination with post- hoc interview and questionnaire data, to offer new insights into the cognitive processes of successful and unsuccessful candidates in a reading test. It also gives unprecedented insights into the cognitive processing of successful and unsuccessful readers doing language tests. As a consequence, the findings should be of value to teachers and learners, and also to examination boards seeking to validate and prepare reading tests, as well as psycholinguists and others interested in the cognitive processes of readers

Impact of L-carnitine and selenium treatment on testicular apoptosis in rats exposed to 2.45 GHz microwave energy

Objective: It has been suggested that electromagnetic radiation (EMR) by wireless devices (2.45 GHz) induces testicular apoptosis. We investigated if supplemental selenium (Se) and L-carnitine may reduce this adverse effect. Material: Twelve-week old maleWistar albino rats were used in this study. Twenty-four rats were equally divided into four groups which were named as: sham group, EMR-only, EMR+L-carnitine (1.5 mg L-carnitine/ kg/day) and EMR+Se (1.5 mg Se/kg/ every other day). Results: The level of Bcl-2, Bax, caspase-3 and -8 were compared and a significant difference was found between the sham and EMR-only groups (p < 0.05), and Bcl-2, Bax, caspase-3 and -8 expressions increased in the EMR-only group. The level of Bcl-2, Bax, tumour necrosis factor-alpha (TNF-α), caspase- 3 and -8 were compared and a significant difference was found between the sham and EMR+L-carnitine groups (p < 0.05) and Bcl-2, Bax, TNF-α, caspase-3 and -8 expressions increased in the EMR+L-carnitine group. The level of Bcl-2, Bax, TNF-α, caspase-3 and -8 were compared and a significant difference was found between the sham and EMR+Se groups (p < 0.05) and Bcl-2, Bax, TNF-α, caspase-3 and -8 expressions increased in the EMR+Se group. When the expression of caspase-8 was compared, a significant difference was found between the EMR-only and EMR+Se groups (p < 0.05). Caspase-8 expression decreased in EMR+Se group compared with EMR-only group. Conclusion: Electromagnetic radiation exposure resulted in testicular apoptosis in rats, mainly by the intrinsic pathways by down-regulated expression of caspase-8. Reduction in the activation of the intrinsic pathway of apoptosis was found higher with selenium administration compared with L-carnitine administration

Accumulation of tissue factor in endothelial cells induces cell apoptosis, mediated through p38 and p53 activation

We previously reported that high levels of tissue factor (TF) can induce cellular apoptosis in endothelial. In this study, TF-mediated mechanisms of induction of apoptosis were explored. Endothelial cells were transfected to express wild-type TF. Additionally, cells were transfected to express Asp253-substituted, or Ala253-substitued TF to enhance or prevent TF release respectively. Alternatively, cells were pre-incubated with TF-rich and TF-poor microvesicles. Cell proliferation, apoptosis and the expression of cyclin D1, p53, bax and p21 were measured following activation of cells with PAR2-agonist peptide. Greatest levels of cell proliferation and cyclin D1 expression were observed in cells expressing wild-type or Asp253-substituted TF. In contrast, increased cellular apoptosis was observed in cells expressing Ala253-substituted TF, or cells pre-incubated with TF-rich microvesicles. The level of p53 protein, p53-phosphorylation at ser33, p53 nuclear localisation and transcriptional activity, but not p53 mRNA, were increased in cells expressing wild-type and Ala253-substituted TF, or in cells pre-incubated with TF-rich microvesicles. However, the expression of bax and p21 mRNA, and Bax protein were only increased in cells pre-incubated with TF-rich microvesicle and in cells expressing Ala253-substituted TF. Inhibition of the transcriptional activity of p53 using pifithrin-α suppressed the expression of Bax. Finally, siRNA–mediated suppression of p38α, or inhibition using SB202190 significantly reduced the p53 protein levels, p53 nuclear localisation and transcriptional activity, suppressed Bax expression and prevented cellular apoptosis. In conclusion, accumulation of TF within endothelial cell, or sequestered from the surrounding can induce cellular apoptosis through mechanisms mediated by p38, and involves the stabilisation of p53