Participation of the cholinergic system in the ethanol-induced suppression of paradoxical sleep in rats

Sleep disturbance is among the many consequences of ethanol abuse in both humans and rodents. Ethanol consumption can reduce REM or paradoxical sleep (PS) in humans and rats, respectively. The first aim of this study was to develop an animal model of ethanol-induced PS suppression. This model administered intragastrically (by gavage) to male Wistar rats (3 months old, 200-250 g) 0.5 to 3.5 g/kg ethanol. The 3.5 g/kg dose of ethanol suppressed the PS stage compared with the vehicle group (distilled water) during the first 2-h interval (0-2 h; 1.3 vs 10.2; P < 0.001). The second aim of this study was to investigate the mechanisms by which ethanol suppresses PS. We examined the effects of cholinergic drug pretreatment. The cholinergic system was chosen because of the involvement of cholinergic neurotransmitters in regulating the sleep-wake cycle. A second set of animals was pretreated with 2.5, 5.0, and 10 mg/kg pilocarpine (cholinergic agonist) or atropine (cholinergic antagonist). These drugs were administered 1 h prior to ethanol (3.5 g/kg) or vehicle. Treatment with atropine prior to vehicle or ethanol produced a statistically significant decrease in PS, whereas pilocarpine had no effect on minutes of PS. Although the mechanism by which ethanol induces PS suppression is not fully understood, these data suggest that the cholinergic system is not the only system involved in this interaction.

Genomic alterations detected by comparative genomic hybridization in ovarian endometriomas

Endometriosis is a complex and multifactorial disease. Chromosomal imbalance screening in endometriotic tissue can be used to detect hot-spot regions in the search for a possible genetic marker for endometriosis. The objective of the present study was to detect chromosomal imbalances by comparative genomic hybridization (CGH) in ectopic tissue samples from ovarian endometriomas and eutopic tissue from the same patients. We evaluated 10 ovarian endometriotic tissues and 10 eutopic endometrial tissues by metaphase CGH. CGH was prepared with normal and test DNA enzymatically digested, ligated to adaptors and amplified by PCR. A second PCR was performed for DNA labeling. Equal amounts of both normal and test-labeled DNA were hybridized in human normal metaphases. The Isis FISH Imaging System V 5.0 software was used for chromosome analysis. In both eutopic and ectopic groups, 4/10 samples presented chromosomal alterations, mainly chromosomal gains. CGH identified 11q12.3-q13.1, 17p11.1-p12, 17q25.3-qter, and 19p as critical regions. Genomic imbalances in 11q, 17p, 17q, and 19p were detected in normal eutopic and/or ectopic endometrium from women with ovarian endometriosis. These regions contain genes such as POLR2G, MXRA7 and UBA52 involved in biological processes that may lead to the establishment and maintenance of endometriotic implants. This genomic imbalance may affect genes in which dysregulation impacts both eutopic and ectopic endometrium.

Kaurene diterpene induces apoptosis in U87 human malignant glioblastoma cells by suppression of anti-apoptotic signals and activation of cysteine proteases

Gliomas are the most common and malignant primary brain tumors in humans. Studies have shown that classes of kaurene diterpene have anti-tumor activity related to their ability to induce apoptosis. We investigated the response of the human glioblastoma cell line U87 to treatment with ent-kaur-16-en-19-oic acid (kaurenoic acid, KA). We analyzed cell survival and the induction of apoptosis using flow cytometry and annexin V staining. Additionally, the expression of anti-apoptotic (c-FLIP and miR-21) and apoptotic (Fas, caspase-3 and caspase-8) genes was analyzed by relative quantification (real-time PCR) of mRNA levels in U87 cells that were either untreated or treated with KA (30, 50, or 70 µM) for 24, 48, and 72 h. U87 cells treated with KA demonstrated reduced viability, and an increase in annexin V- and annexin V/PI-positive cells was observed. The percentage of apoptotic cells was 9% for control cells, 26% for cells submitted to 48 h of treatment with 50 µM KA, and 31% for cells submitted to 48 h of treatment with 70 µM KA. Similarly, in U87 cells treated with KA for 48 h, we observed an increase in the expression of apoptotic genes (caspase-8, -3) and a decrease in the expression of anti-apoptotic genes (miR-21 and c-FLIP). KA possesses several interesting properties and induces apoptosis through a unique mechanism. Further experiments will be necessary to determine if KA may be used as a lead compound for the development of new chemotherapeutic drugs for the treatment of primary brain tumors.

Chromogenic in situ hybridization compared with other approaches to evaluate HER2/neu status in breast carcinomas

Human epidermal growth factor receptor 2 (HER2) has been evaluated in breast cancer patients to identify those most likely to benefit from herceptin-targeted therapy. HER2 amplification, detected in 20-30% of invasive breast tumors, is associated with reduced survival and metastasis. The most frequently used technique for evaluating HER2 protein status as a routine procedure is immunohistochemistry (IHC). HER2 copy number alterations have also been evaluated by fluorescence in situ hybridization (FISH) in moderate immunoexpression (IHC 2+) cases. An alternative procedure to evaluate gene amplification is chromogenic in situhybridization (CISH), which has some advantages over FISH, including the correlation between HER2 status and morphological features. Other methodologies have also been used, such as silver-enhanced in situ hybridization (SISH) and quantitative real-time RT-PCR, to determine the number of HER2 gene copies and expression, respectively. Here we will present a short and comprehensive review of the current advances concerning HER2 evaluation in human breast cancer.

Lettre à un ami sur la suppression de l'habit ecclésiastique, & sur le danger que court le clergé de France, de pertes plus réelles ([Reprod.])

Collection : Les archives de la Révolution française ; 8.262

Edit du Roy, portant suppression des offices de contrôlleurs des bans de mariage qui avoient esté créez par l'Edit du mois de septembre 1697 . Et tarif des droits que Sa Majesté veut estre payez à l'avenir pour le contrôlle & l'enregistrement de chacune publication de ban de mariage conformement audit Edit. Donné à Versailles au mois de mars 1702. Registré en Parlement le 26 avril 1702.

[Acte royal. 1702-03-00. Versailles]

Arrêt du conseil d'Etat qui commet J.-B. Dumont pour l'exécution de l'Edit de janvier 1717, portant suppression des offices de contrôleurs des greffiers garde-minutes des chancelleries près les cours et sièges présidiaux

[Acte. 1717-02-20. Paris]

Édit... portant suppression des offices de conseillers du Roy notaires et secrétaires dans les cours de Parlement, Chambres des Comptes et Cours des Aydes, Grand Conseil, Requestes de l'Hostel et du Palais, Conseils supérieurs et provinciaux, Cours des monnoies et Bureaux des Finances ; ordonne que les greffiers en chef des cours seront tous tenus de se pourvoir de l'un des offices de secrétaires, créez dans les chancelleries près les Cours par édit... de juin 1715 ; que la vente des offices de receveurs et payeurs des gages des officiers desdites chancelleries créez par édits... de novembre 1707 et décembre 1708 sera continuée ; et restablit les gardes des sceaux des chancelleries près les Cours dans leurs offices, en payant un supplément de finance... [Enregistré au Parlement le 13 juin 1716.]

[Acte royal. 1716-05-00. Paris]

Etat général des dettes de l'Etat à la mort du feu Roy Louis XIV . Arrivée le premier septembre 1715. Leur réduction & payement. Avec la suppression & extinction d'un grand nombre d'offices & droits, à la décharge des peuples, jusqu'au premier septembre 1720

[Acte. 1720]

Arrêt du conseil d'état portant suppression d'un livre imprimé sans permission

[Acte. 1752-06-28. Versailles]

(Arrêt du Grand Conseil qui ordonne la suppression d'un libelle intitulé : "Mémoire pour Messire Gui-Michel Billard de Lorière, conseiller au Grand Conseil, contre dame Marie-Henriette de Saint-Simon, son épouse.)

[Acte. 1755-08-30. Paris]