TAF15 is important for cellular proliferation and regulates the expression of a subset of cell cycle genes through miRNAs.

TAF15 (formerly TAFII68) is a member of the FET (FUS, EWS, TAF15) family of RNA- and DNA-binding proteins whose genes are frequently translocated in sarcomas. By performing global gene expression profiling, we found that TAF15 knockdown affects the expression of a large subset of genes, of which a significant percentage is involved in cell cycle and cell death. In agreement, TAF15 depletion had a growth-inhibitory effect and resulted in increased apoptosis. Among the TAF15-regulated genes, targets of microRNAs (miRNAs) generated from the onco-miR-17 locus were overrepresented, with CDKN1A/p21 being the top miRNAs-targeted gene. Interestingly, the levels of onco-miR-17 locus coded miRNAs (miR-17-5p and miR-20a) were decreased upon TAF15 depletion and shown to affect the post-transcriptional regulation of TAF15-dependent genes, such as CDKN1A/p21. Thus, our results demonstrate that TAF15 is required to regulate gene expression of cell cycle regulatory genes post-transcriptionally through a pathway involving miRNAs. The findings that high TAF15 levels are needed for rapid cellular proliferation and that endogenous TAF15 levels decrease during differentiation strongly suggest that TAF15 is a key regulator of maintaining a highly proliferative rate of cellular homeostasis.

Problematique de la prise en charge de l'infarctus du myocarde a L'Hopital Desegou (Mali)

INTRODUCTION: Myocardial infarction is a major emergency involving life-threatening in the absence of appropriate treatment. The aim of this study was to analyze the problem of management of myocardial infarction in a second reference hospital in Mali. PATIENTS AND METHODS : This was a prospective descriptive study over a period of six months from January to June 2010. It concerned all patients admitted for myocardial infarction in intensive care.The diagnosis was suspected in chest pain or the occurrence of complications (PAO, cardiogenic shock) and electrocardiogram signs on at least two precordial leads. The parameters studied were: age,reason for admission, risk factors, hemodynamic parameters, the deadline for completion of the ECG, the topography of lesions and electrical changes within 15 days. RESULTS: A male was found with a mean age of 54.62 years. Chest pain was the main reason for admission (6 cases) followed by cardiogenic shock (1 case) and acute pulmonary edema (1 case). The electrocardiogram was performed in 7 patients more than 24 hours after admission. The anterior territory was the most affected. On admission three patients had a systolic pressure below 90 mmHg.The evolution was marked by occurred heart failure (3 cases) and death (2 cases). CONCLUSION: The lack of diagnostic and therapeutic method in our heath facility helps to increase morbidity and mortality associated with myocardial infarction.

[Problems with the management of myocardial infarction at the Desegou Hospital]. / Problematique de la prise en charge de l'infarctus du myocarde a L'Hopital Desegou (Mali)

INTRODUCTION: Myocardial infarction is a major emergency involving life-threatening in the absence of appropriate treatment. The aim of this study was to analyze the problem of management of myocardial infarction in a second reference hospital in Mali. PATIENTS AND METHODS: This was a prospective descriptive study over a period of six months from January to June 2010. It concerned all patients admitted for myocardial infarction in intensive care.The diagnosis was suspected in chest pain or the occurrence of complications (PAO, cardiogenic shock) and electrocardiogram signs on at least two precordial leads. The parameters studied were: age,reason for admission, risk factors, hemodynamic parameters, the deadline for completion of the ECG, the topography of lesions and electrical changes within 15 days. RESULTS: A male was found with a mean age of 54.62 years. Chest pain was the main reason for admission (6 cases) followed by cardiogenic shock (1 case) and acute pulmonary edema (1 case). The electrocardiogram was performed in 7 patients more than 24 hours after admission. The anterior territory was the most affected. On admission three patients had a systolic pressure below 90 mmHg.The evolution was marked by occurred heart failure (3 cases) and death (2 cases). CONCLUSION: The lack of diagnostic and therapeutic method in our heath facility helps to increase morbidity and mortality associated with myocardial infarction.

Gene expression profile and response to trastuzumab-docetaxel-based treatment in breast carcinoma.

BACKGROUND: Resistance to trastuzumab is often observed in women with human epidermal growth factor receptor 2 (HER2)-positive breast cancer and has been shown to involve multiple potential mechanisms. We examined the ability of microarray analyses to determine the potential markers of pathological complete response (pCR). METHODS: We conducted an analysis of tumours from 38 patients with locally advanced HER2-positive breast cancer who had received trastuzumab combined with docetaxel. Quantitative reverse transcriptase (RT)-PCR was used to assess the expression of 30 key genes; microarray analyses were carried out on 25 tumours to identify a prognostic gene expression profile, with 13 blinded samples used to validate the identified profile. RESULTS: No gene was found to correlate with response by RT-PCR. The microarray analysis identified a gene expression profile of 28 genes, with 12 upregulated in the pCR group and 16 upregulated in non-pCR. The leave-one-out cross-validation test exhibited 72% accuracy, 86% specificity, and 55% sensitivity. The 28-gene expression profile classified the 13 validation samples with 92% accuracy, 89% specificity, and 100% sensitivity. CONCLUSION: Our results suggest that genes not involved in classical cancer pathways such as apoptosis or DNA repair could be involved in responses to a trastuzumab-docetaxel-based regimen. They also describe for the first time a gene expression signature that predicts trastuzumab response.

Transcriptome analysis identifies genes with enriched expression in the mouse central extended amygdala.

The central extended amygdala (EAc) is an ensemble of highly interconnected limbic structures of the anterior brain, and forms a cellular continuum including the bed nucleus of the stria terminalis (BNST), the central nucleus of the amygdala (CeA) and the nucleus accumbens shell (AcbSh). This neural network is a key site for interactions between brain reward and stress systems, and has been implicated in several aspects of drug abuse. In order to increase our understanding of EAc function at the molecular level, we undertook a genome-wide screen (Affymetrix) to identify genes whose expression is enriched in the mouse EAc. We focused on the less-well known BNST-CeA areas of the EAc, and identified 121 genes that exhibit more than twofold higher expression level in the EAc compared with whole brain. Among these, 43 genes have never been described to be expressed in the EAc. We mapped these genes throughout the brain, using non-radioactive in situ hybridization, and identified eight genes with a unique and distinct rostro-caudal expression pattern along AcbSh, BNST and CeA. Q-PCR analysis performed in brain and peripheral organ tissues indicated that, with the exception of one (Spata13), all these genes are predominantly expressed in brain. These genes encode signaling proteins (Adora2, GPR88, Arpp21 and Rem2), a transcription factor (Limh6) or proteins of unknown function (Rik130, Spata13 and Wfs1). The identification of genes with enriched expression expands our knowledge of EAc at a molecular level, and provides useful information to toward genetic manipulations within the EAc.

Mu-opioid receptor activation induces transcriptional plasticity in the central extended amygdala.

Addiction develops from the gradual adaptation of the brain to chronic drug exposure, and involves genetic reprogramming of neuronal function. The central extended amygdala (EAc) is a network formed by the central amygdala and the bed nucleus of the stria terminalis. This key site controls drug craving and seeking behaviors, and has not been investigated at the gene regulation level. We used Affymetrix microarrays to analyze transcriptional activity in the murine EAc, with a focus on mu-opioid receptor-associated events because these receptors mediate drug reward and dependence. We identified 132 genes whose expression is regulated by a chronic escalating morphine regimen in the EAc from wild-type but not mu-opioid receptor knockout mice. These modifications are mostly EAc-specific. Gene ontology analysis reveals an overrepresentation of neurogenesis, cell growth and signaling protein categories. A separate quantitative PCR analysis of genes in the last of these groups confirms the dysregulation of both orphan (Gpr88) and known (DrD1A, Adora2A, Cnr1, Grm5, Gpr6) G protein-coupled receptors, scaffolding (PSD95, Homer1) and signaling (Sgk, Cap1) proteins, and neuropeptides (CCK, galanin). These transcriptional modifications do not occur following a single morphine injection, and hence result from long-term adaptation to excessive mu receptor activation. Proteins encoded by these genes are classically associated with spine modules function in other brain areas, and therefore our data suggest a remodeling of EAc circuits at sites where glutamatergic and monoaminergic afferences interact. Together, mu receptor-dependent genes identified in this study potentially contribute to drug-induced neural plasticity, and provide a unique molecular repertoire towards understanding drug craving and relapse.

Gene expression is altered in the lateral hypothalamus upon activation of the mu opioid receptor.

The lateral hypothalamus (LH) is a brain structure that controls hedonic properties of both natural rewards and drugs of abuse. Mu opioid receptors are known to mediate drug reward, but whether overstimulation of these receptors impacts on LH function has not been studied. Here we have used a genome-wide microarray approach to identify LH responses to chronic mu opioid receptor activation at the transcriptional level. We have subjected wild-type and mu opioid receptor knockout mice to an escalating morphine regimen, which produces severe physical dependence in wild-type but not mutant animals. We have analyzed gene profiles in LH samples using the 430A.2 Affymetrix array and identified a set of 25 genes whose expression is altered by morphine in wild-type mice only. The regulation was confirmed for a subset of these genes using real-time quantitative PCR on samples from independent treatments. Altered expression of aquaporin 4, apolipoprotein D, and prostaglandin synthase is indicative of modified LH physiology. The regulation of two signaling genes (the serum glucocorticoid kinase and the regulator of G protein signaling 4) suggests that neurotransmission is altered in LH circuitry. Finally, the downregulation of apelin may indicate a potential role for this neuropeptide in opioid signaling and hedonic homeostasis. Altogether, our study shows that chronic mu opioid receptor stimulation induces gene expression plasticity in the LH and provides a unique collection of mu opioid receptor-dependent genes that potentially contribute to alter reward processes in addictive diseases.

Bcl2, a transcriptional target of p38alpha, is critical for neuronal commitment of mouse embryonic stem cells.

Mouse embryonic stem (ES) cells remain pluripotent in vitro when grown in the presence of leukemia inhibitory factor (LIF) cytokine. LIF starvation leads to cell commitment, and part of the ES-derived differentiated cells die by apoptosis together with caspase3-cleavage and p38alpha activation. Inhibition of p38 activity by chemical compounds (PD169316 and SB203580), along with LIF withdrawal, leads to different outcomes on cell apoptosis, giving the opportunity to study the influence of apoptosis on cell differentiation. By gene profiling studies on ES-derived differentiated cells treated or not with these inhibitors, we have characterized the common and specific set of genes modulated by each inhibitor. We have also identified key genes that might account for their different survival effects. In addition, we have demonstrated that some genes, similarly regulated by both inhibitors (upregulated as Bcl2, Id2, Cd24a or downregulated as Nodal), are bona fide p38alpha targets involved in neurogenesis and found a correlation with their expression profiles and the onset of neuronal differentiation triggered upon retinoic acid treatment. We also showed, in an embryoid body differentiation protocol, that overexpression of EGFP (enhanced green fluorescent protein)-BCL2 fusion protein and repression of p38alpha are essential to increase formation of TUJ1-positive neuronal cell networks along with an increase in Map2-expressing cells.

Retinoic acid induces TGFbeta-dependent autocrine fibroblast growth.

To evaluate the role of murine TFIID subunit TAF4 in activation of cellular genes by all-trans retinoic acid (T-RA), we have characterized the T-RA response of taf4(lox/-) and taf4(-/-) embryonic fibroblasts. T-RA regulates almost 1000 genes in taf4(lox/-) cells, but less than 300 in taf4(-/-) cells showing that TAF4 is required for T-RA regulation of most, but not all cellular genes. We further show that T-RA-treated taf4(lox/-) cells exhibit transforming growth factor (TGF)beta-dependent autocrine growth and identify a set of genes regulated by loss of TAF4 and by T-RA corresponding to key mediators of the TGFbeta signalling pathway. T-RA rapidly and potently induces expression of connective tissue growth factor (CTGF) via a conserved DR2 type response element in its proximal promoter leading to serum-free autocrine growth. These results highlight the role of TAF4 as a cofactor in the cellular response to T-RA and identify the genetic programme of a novel cross talk between the T-RA and TGFbeta pathways that leads to deregulated cell growth.

Apoptosis and differentiation commitment: novel insights revealed by gene profiling studies in mouse embryonic stem cells.

Mouse embryonic stem (ES) cells remain pluripotent in vitro when grown in the presence of leukemia inhibitory factor (LIF). LIF starvation leads to apoptosis of some of the ES-derived differentiated cells, together with p38alpha mitogen-activated protein kinase (MAPK) activation. Apoptosis, but not morphological cell differentiation, is blocked by a p38 inhibitor, PD169316. To further understand the mechanism of action of this compound, we have identified its specific targets by microarray studies. We report on the global expression profiles of genes expressed at 3 days upon LIF withdrawal (d3) compared to pluripotent cells and of genes whose expression is modulated at d3 under anti-apoptotic conditions. We showed that at d3 without LIF cells express, earlier than anticipated, specialized cell markers and that when the apoptotic process was impaired, expression of differentiation markers was altered. In addition, functional tests revealed properties of anti-apoptotic proteins not to alter cell pluripotency and a novel role for metallothionein 1 gene, which prevents apoptosis of early differentiated cells.

Head and neck squamous cell carcinoma transcriptome analysis by comprehensive validated differential display.

Head and neck squamous cell carcinoma (HNSCC) is common worldwide and is associated with a poor rate of survival. Identification of new markers and therapeutic targets, and understanding the complex transformation process, will require a comprehensive description of genome expression, that can only be achieved by combining different methodologies. We report here the HNSCC transcriptome that was determined by exhaustive differential display (DD) analysis coupled with validation by different methods on the same patient samples. The resulting 820 nonredundant sequences were analysed by high throughput bioinformatics analysis. Human proteins were identified for 73% (596) of the DD sequences. A large proportion (>50%) of the remaining unassigned sequences match ESTs (expressed sequence tags) from human tumours. For the functionally annotated proteins, there is significant enrichment for relevant biological processes, including cell motility, protein biosynthesis, stress and immune responses, cell death, cell cycle, cell proliferation and/or maintenance and transport. Three of the novel proteins (TMEM16A, PHLDB2 and ARHGAP21) were analysed further to show that they have the potential to be developed as therapeutic targets.

[Sacral root neuromodulation for the treatment of urinary incontinence reported to detrusor hyperactivity]. / Traitement de l'incontinence urinaire secondaire à une hyperactivité vésicale par neuromodulation sacrée.

BACKGROUND AND PURPOSE: This paper reviews therapeutic sacral neuromodulation for treating urinary urge incontinence related to detrusor hyperactivity. METHODS: We reported data from our department and from the international literature on topics such as the physiological basis of neuromodulation, techniques of testing, chronic implantation and clinical results. RESULTS: In an intention to treat analysis , neuromodulation results varied from 21.5 to 25% globally. On implanted patients, the response rate varied from 40 to 88% and was stable. Sub-chronic test morbidity was very rare. Surgical revision rate was reported from 6.25 to 37.7%. CONCLUSIONS: Neuromodulation strongly ameliorates approximately a third of all the patients with urge urinary incontinence due to detrusor hyperactivity. New technical improvements should lead to better results in the future.

[Tolerance of ivermectin treatment of rural communities infected by savannah onchocerciasis in Mali]. / Tolérance d'un traitement de collectivités rurales atteintes d'onchocercose de savane par l'ivermectine au Mali.

In an open clinical trial in phase IV, 856 onchocerciasis infected subjects received 150 micrograms/kg of ivermectin in May 1987. While 607 were included as witness. This cohort was revisited 7 and 12 months after. In June 1988, the same treatment was administrated to the previously treated subjects, and the witnesses received their first ivermectin' dose. The clinical tolerance of the treatment appears good and, even improved during the second dose one year after. Among the subjects treated in May 1987, 15.2% of them showed secondary reactions mostly discrete or moderate, precocious and quickly reversible after a second dose. Only 8 of them were incommodated in their daily occupations. A second treatment of these same subjects one year later, caused reactions of feeble intensity 3.7% only. The research of intolerance risk factors, incriminated the high density of microfilaremia. This incite to be careful in mass treatment of hyperendemic area.