Evaluation of genetic diversity in Sulla coronaria from different geographical populations in Tunisia by inter simple sequence repeat (ISSR)

Five oligonucleotides generating 116 markers complementary to simple sequence repeats were used in order to characterize wild and cultivated ecotypes of Sulla coronaria and assess genetic diversity suitable in breeding programs. While analysing populations, a large genetic variability was revealed and supported by the preferentially allogamous mating system of the species. Moreover, the highest level of intra-population variations (Hpop/Hsp = 69.9) either of wild or cultivated accessions have been strongly evidenced by a significant adaptation to variety of habitats. In addition, the structure of populations was independent from the bioclimatic stages and was not affected by environmental factors as shown by the non correlation between the geographic and the Nei and Li’s genetic distances (r= 0.461 and p=0.068>0.05). The unweighted pair group method with arithmetic mean (UPGMA) genetic relationships showed that some local spontaneous accessions characterised by an orthotropic port (Jebel Zit and Beja) were also molecularly similar to other cultivars.Key words: Sulla coronaria, wild and cultivated forms, intra and inter-populations variability, microsatellites (ISSRs), bioclimatic stages

Cytokeratin 18 expression pattern correlates with renal cell carcinoma progression: Relationship with Snail

Renal cell carcinoma (RCC) is the most common type of kidney cancer and recent developments in the molecular biology of RCC have identified multiple pathways associated with the development of this cancer. This study aimed at analyzing the expression pattern of cytokeratin 18 (CK18) in RCC patients and its prognostic relevance. We quantified CK18 mRNA expression and protein using real-time reverse transcription quantitative polymerase chain reaction (RT-QPCR) and immunohistochemistry, respectively, in paired tumor and non-tumor samples from 42 patients. Our data indicate that CK18 mRNA and proteins levels increased with advanced stage and grade of the disease. Using primary (RCC5) and metastatic renal cell carcinoma (RCC5 met) cell lines, we demonstrated that CK18 expression was 5-fold higher in the metastatic as compared to the primary RCC cell line and correlated with a migratory phenotype characterized by a distinct elongated morphology as revealed by Phalloidin staining. In addition, RCC5 met cells displayed an increased capacity to attach to fibronectin and collagen which was lost following CK18 knock-down. Our data also indicate that the expression of CK18 was associated with increased Snail expression which correlated positively with advanced disease in RCC patients. The present findings suggest that CK18 may play an important role in the progression of RCC and it may be used as a new predictor for RCC

Impact of Smoking intensity on Outcomes of Patients with Non Muscle Invasive Bladder Cancer Treated by BCG Immunotherapy.

International audienceAbstract Background: Cigarette smoking is a well-known risk factor of bladder carcinogenesis. The clinical impact of smoking on bladder cancer recurrence and response to BCG immunotherapy remains unclear. We sought to investigate the effect of smoking intensity on bladder cancer response to BCG therapy, and the interactions between smoking and clinicopathological factors on bladder cancer recurrence. Methods: Clinical information was obtained from 81 smokers patients (smokers at diagnosis) with NMIBC treated with transurethral resection of the bladder tumor followed by BCG immunotherapy. The distribution of smoking intensity on patient age (≥60 years or <60 years), gender, tumor grade, tumor stage, carcinoma in situ, multiplicity and tumor size was assessed. The effect of cigarette smoking on cancer recurrence was estimated using Cox proportional hazard models and Kaplan-Meier analysis. Results: The results showed that smoking intensity was significantly associated with response to BCG immunotherapy (p = 0.010). Univariate Cox regression analysis of clinicopathologic characteristics showed that PT1 stage, tumor size more than 3 cm and smoking intensity significantly increased the risk of recurrence (respectively, p = 0.006; p = 0.008 and p = 0.012). These results were confirmed by Kaplan-Meier survival curves. In addition, multivariate analysis using Cox regression selected the model involving stage, tumor size and smoking intensity as the quasi-independent predictor of recurrence.Conclusion: These findings suggest that cigarette smoking is an independent predictor for patients with NMIBC. Although the current evidence supports a positive link between smoking intensity and the risk of recurrence on NMIBC treated by BCG immunotherapy, additional studies, are needed before definitive conclusions can be drawn

Hypericum Humifusum Leaves Attenuates Hepatic Ischemia-Reperfusion Injury in a Rat Model

Introduction. Effective prevention strategies require specific actions during the different phases of ischemia-reperfusion (I-R) injury. The objective of our study is to evaluate the effect of aqueous extract of Hypericum humifusum leaves (HHL) on liver I-R model in Rat.Material and methods. Animals were subjected to 90 min of hepatic ischemia followed by reperfusion (120 min). HHL extract (25 mg/mL/kg) was injected 15 min before reperfusion. To evaluate the effect of HHL extract on I-R, we have monitored transaminases levels, Malondialdehyde (MDA) concentration, histological lesions (apoptosis and necrosis) and compared the results to a reference oxidant vitamin E.Results. The determination of total phenol extracts of HHL was 59.91 ± 0.35 mg of Gallic Acid/g dry plant material with higher antioxidant activity (91.73% ± 1.67) compared to vitamin E (87.42%). Using aqueous extract of HHL, we noted a significant decrease of AST and ALT [1129 UI (585/1995) and 768 UI (335/1375)] compared to no-treated group [5,585.5 UI (5,035/12,070) and 8,099.5 UI (5,040/12,326)] as a decrease in MDA content [85.7% protection (50.9/91.5)]. HHL extract reduce the damage induced by I-R of 48.7% (27/48.7) and 96.1% (95.7/96.5) for necrosis and apoptosis lesions respectively.Conclusion. HHL aqueous extract have potential to protect liver from the damage effect induced by I-R better than vitamin E solution

La valeur pronostique du PASS score dans la distinction entre phéochromocytome bénin et malin

International audienceBACKGROUND: Differentiating malignant from benign pheochromocytoma has been challenging when based on histologic features. This is due to the definition of malignant pheochromocytoma which are defined by the presence of metastases. A PASS score was developed and according to many authors, a PASS score> =4 identified potentially malignant tumors.AIM: To assess the prognostic value of PASS score in differentiating benign pheochromocytomas from malignant ones.METHODS: The records of 11 patients with tumors diagnosed as "pheochromocytoma" were identified from 1970 to 2010 in the files of the pathology, intern medicine and biochemistry departments of the Charles Nicolle hospital and Pasteur Institute. Receiver operating characteristics (ROC) curve analysis was performed to evaluate the diagnostic performance of PASS. The logistic model was developed using the 11 predictive variables. Its performance was evaluated by calculating the area under the ROC curve and comparing it with that of the PASS.RESULTS: In benign tumors, The PASS score was =4 in 6 cases. In malignant tumors, the PASS score was >=4 in both cases. According to the ROC curve analysis, a PASS equal or superior to 4 identifies malignant pheochromocytoma with a sensitivity of 50% and a specificity of 45%.CONCLUSION: I think that PASS score, despite its low sensitivity, may help to reserve the more aggressive treatment and narrow follow up for potentially malignant tumors. Widespread of this called score with complete clinical data will help to validate these findings and to add other prognostic factors of value that could be a part of this scaled score such as immunohistochemical findings.Prérequis : La distinction entre phéochromocytome bénin et malin peut se révéler délicate sur le plan morphologique. Cette difficulté découle de la définition même des phéochromocytomes malins dont le diagnostic repose sur la présence de métastases. Un score de PASS a été développé afin de contribuer à cette distinction. Un score de PASS >4 est corrélé à un haut potentiel de malignité.But: Etudier la valeur pronostique du score de PASS afin de différencier les phéochromocytomes bénins et les phéchromocytomes malins.méthodes: Nous rapportons une étude à propos de 11 patients présentant des phéochromocytomes diagnostiqués entre 1970 et 2010. Des courbes ROC ont été utilisées afin d’évaluer le potentiel diagnostique du score de PASS. Ce modèle logistique a été développé en utilisant 11 variables prédictives. Sa performance a été évaluée en calculant l’aire comprise sous la courbe ROC.résultats: Dans les tumeurs bénignes, le sore de PASS était <4 dans 3 cas et ≥4 dans 6 cas. Dans les tumeurs malignes, le score de PASS était ≥4 dans les 2 cas. L’analyse de la courbe ROC a permis d’établir qu’un score ≥ à 4 permettait d’identifier les phéochromocytomes malins avec une sensibilité de 50% et une spécificité de 45%.Conclusion: Le score de PASS peut contribuer au diagnostic des phéochromocytomes malins et ce malgré sa faible sensibilité.Cependant, ce score pourrait être enrichi par de nouveaux items tel que les données imunohistochimiques permettant ainsi d’améliorer sa sensibilité

Therapeutic effect of flaxseed oil on experimental pulmonary fibrosis induced by bleomycin in rats

Pulmonary fibrosis is a chronic progressive disorder in which excessive deposition of extracellular matrix leads to irreversible scarring to interstitial lung tissue. In this study, we search to evaluate the therapeutic effect of flaxseed oil (FO) in experimental bleomycin (BLM)-induced pulmonary fibrosis. During our study, 30 male Wistar rats (weight range, 180–220 g) were divided into three groups: the control group (W) received no treatment; the second group (C) received BLM; and the third group (T) received BLM and FO for 21 days. Metabolites present in the bronchoalveolar lavage fluid (BALF) marking the changes obtained following treatment with FO were determined, histological changes in the lungs were evaluated, fatty acids present in lungs and erythrocytes of rats groups were determined by gas chromatography, and oxidative stress and antioxidant enzyme activity in the lung tissue were also recorded. Our results displayed that rat body weight decreased while fibrosis score and inflammatory index in lung tissue were significantly increased after bleomycin instillation. Administration of bleomycin followed by FO treatment reduced bleomycin-induced weight loss, increased proline, glucose, and glycerid rates in BALF and which are characterized by their anti-inflammatory effect and confirming the histological results proved by a decrease in inflammatory index and fibrosis score. This oil also significantly reduced thiobarbitunic acid reactive substance levels in the lungs of rats and increased levels of SOD and CAT and increased fatty acids levels promoting anti-inflammatory reactions especially in erythrocytes (linoleic, arachidonic, docosapentaenoic, and dihomo-γ-linoleic acids). In conclusion, these findings indicate that FO treatment significantly attenuated the increased pulmonary damage induced by bleomycin

Prognostic value of Bcl-2 and Bax tumor cell expression in patients with non muscle-invasive bladder cancer receiving bacillus Calmette-Guerin immunotherapy.

International audienceApoptosis is the distinctive form of programmed cell death that complements cell proliferation in maintaining normal tissue homeostasis. The significance of constitutive apoptosis in the recurrence of Non Muscle Invasive Bladder Cancer has yet to be investigated. The aim of this study is to investigate the prognostic significance of Bax and Bcl-2 in terms of recurrence after BCG immunotherapy. Immunohistochemical analysis was performed on frozen biopsies to evaluate bcl-2 and Bax proteins expression in 28 cases of NMIBC. All patients with confirmed NMIBC were treated with intravesical BCG-immunotherapy. The follow up was performed for 26 months. The correlation between clinicopathological, immunohistochemical data and the response to BCG therapy was performed. Univariate analysis showed that, PT1 stage, High grade and Bax expression increased significantly the risk of recurrence (P = 0.015, P = 0.015 and P= 0.034 respectively). In addition, multivariate analysis selected the model involving stage, age, Bax and Bcl-2 expression as the best independent variables of recurrence. In conclusion, the expression of Bcl-2 and Bax in NMIBC could have a prognostic value in assessing the risk of recurrence after BCG immunotherapy. These findings require further investigations on larger cohort in order to ascertain new molecular markers of the response to BCG immunotherapy

Impaired Control of Effector T Cells by Regulatory T Cells: A Clue to Loss of Oral Tolerance and Autoimmunity in Celiac Disease?

International audienceOBJECTIVES:Regulatory T cells (Tregs) are instrumental for tolerance to self-antigens and dietary proteins. We have previously shown that interleukin (IL)-15, a cytokine overexpressed in the intestine of patients with celiac disease (CD), does not impair the generation of functional Tregs but renders human T cells resistant to Treg suppression. Treg numbers and responses of intestinal and peripheral T lymphocytes to suppression by Tregs were therefore compared in CD patients and controls.METHODS:Intraepithelial lymphocytes (IELs) and lamina propria lymphocytes (LPLs) were isolated from duodenal biopsy specimens of CD patients and controls. Concomitantly, CD4+CD25+ T lymphocytes (Tregs) were purified from blood. Responses of IELs and of LPLs, and peripheral lymphocytes (PBLs) to suppression by Tregs were tested by analyzing anti-CD3-induced proliferation and interferon (IFN)-γ production in the presence or absence of peripheral Tregs. Lamina propria and peripheral CD4+CD25+FOXP3+ T cells were assessed by flow cytometry.RESULTS:Although percentages of CD4+CD25+FOXP3+ LPLs were significantly increased in patients with active CD, proliferation and IFN-γ production of intestinal T lymphocytes were significantly less inhibited by autologous or heterologous Tregs in CD patients than in controls (P<0.01). In all tested CD patients, IEL were unable to respond to Tregs. Resistance of LPLs and PBLs to Treg suppression was observed in patients with villous atrophy who had significantly enhanced serum levels of IL-15 compared with patients without villous atrophy and controls.CONCLUSIONS:Our results indicate that effector T lymphocytes from active CD become resistant to suppression by Tregs. This resistance might cause loss of tolerance to gluten, but also to self-antigens.Am J Gastroenterol advance online publication, 22 November 2011; doi:10.1038/ajg.2011.397