Effects of a dopamine agonist (cu 32-085) on rat leydig cells: functional and morphological studies.

The dopamine agonist, 8a-aminoergoline (CU 32-085) (DA) developed for use in the treatment of hyperprolactinaemia and Parkinson's disease induces Leydig cell tumours in rats treated for 21/2 years. The aim of the present study was to investigate the changes in Leydig cell function and growth at earlier stages of treatment with the DA which may contribute to the formation of the Leydig cell tumours. Sprague Dawley rats were treated with CU 32-085 (2mg/kg body weight/day) for 1, 5, 12 or 57 weeks. The Leydig cells from rats treated for 5 weeks produced lower levels of testosterone in response to luteinizing hormone (LH) compared with the controls. The defect in steroidogenesis was associated with a decrease in the numbers of LH receptors and cyclic AMP production, a lesion in steroidogenesis at the 17a-hydroxylase and 17-20 lyase, and an increase in aromatase activity. Histological examination of testis sections from controls and treated rats revealed the presence of atypical sites of Leydig cells surrounded by clusters of macrophages in the testes from the 57 but not the 5 week treated animals. There was 42% and 31% increase in the number of Leydig cells and macrophages respectively, compared with the controls. A method for separating Leydig cells from macrophages was developed and heterogeneity of the Leydig cells was demonstrated. Preliminary evidence indicates that CU 32-085 may selectively inhibit the function of the Leydig cells with lower sedimentation velocities. The effect(s) exerted by CU 32-085 on Leydig cells may result from the increase in circulating levels of LH, ACTH/ glucocorticoids and the decrease in prolactin. Evidence for this was obtained from in vivo studies with human chorionic gonadotrophin (hCG) and dexamethasone which showed an effect similar to that of CU 32-085. LH-stimulated testosterone production was inhibited in the hCG-treated rats and dexamethasone caused a further decrease. The development of Leydig cell tumours caused by treatment with CU 32-085 could be due to the changes in testicular levels of testosterone and oestradiol- 1713 which may result in changes in factors involved in the control of Leydig cell growth. It is concluded that the changes in the biochemical properties of the Leydig cells in vitro after treatment with CU 32-085 provide a sensitive system for detecting the early effects of this compound on Leydig cell function and growth

STRATEGI HUMAS PT PERKEBUNAN NUSANTARA V DALAM MENERAPKAN CORPORATE CULTURE DIKALANGAN KARYAWAN

ABSTRAK Nama : Dirami Pratiwi NIM : 11643202451 Judul : Strategi Humas PT Perkebunan Nusantara V Dalam Menerapkan Corporate Culture Dikalangan Karyawan Coporate Culture berperan penting dalam perkembangan serta kemajuan perusahaan yang menjadi nilai-nilai dasar sikap dan perilaku karyawan di sebuah perusahaan. PT Perkebunan Nusantara V merupakan salah satu perusahaan Badan Usaha Milik Negara yang merupakan anak perusahaan dari PT Perkebunan Nusantara Group. PT Perkebunan Nusantara Group pada saat ini menerapkan SIPro (Sinergi Integritas dan Profesional) sebagai transformasi budaya baru kepada seluruh anak perusahaan yang berada dibawah naungan PT Perkebunan Nusantara Group. Tujuan penelitian untuk mengetahui bagaimana strategi humas PT Perkebunan Nusantara V dalam menerapkan corporate culture dikalangan karyawan dengan metode kualitatif. Peneliti melakukan penelitian di kantor pusat PT Perkebunan Nusantara V Pekanbaru. Informan penelitian ini sebanyak lima orang. Teknik pengumpulan datanya adalah wawancara, observasi dan dokumentasi. Waktu penelitian dilakukan pada bulan November 2019 sampai dengan Januari 2020. Penelitian ini menggunakan teori perencanaan strategi humas Cutlip- Center-Broom. Temuan penelitian ini menghasilkan bahwa strategi humas PT Perkebunan Nusantara V dalam menerapkan corporate culture dikalangan karyawan yaitu Pertama, membuat keputusan mengenai sasaran kepada seluruh karyawan dan tujuan penerapan coporate culture. Kedua, melakukan identifikasi khalayak penentu yaitu kepada Tim Sumber daya manusia. Ketiga, menetapkan kebijakan atau aturan untuk memutuskan strategi yang akan dipilih dengan berpedoman kepada peraturan kerja bersama. Keempat, memutuskan strategi yang akan digunakan dengan menerapkan corporate culture yakni SIPro,berdasarkan hasil penelitian peneliti dilapangan bahwa SIPro sebagai corporate culture yang baru berdampak baik kepada nilai-nilai serta perilaku para karyawan dalam bekerja. Kata Kunci : Strategi, Humas, Corporate Cultur

Essential role of CFTR in PKA-dependent phosphorylation, alkalinization, and hyperpolarization during human dperm capacitation

Mammalian sperm require to spend a limited period of time in the female reproductive tract to become competent to fertilize in a process called capacitation. It is well established that HCO3 − is essential for capacitation because it activates the atypical soluble adenylate cyclase ADCY10 leading to cAMP production, and promotes alkalinization of cytoplasm, and membrane hyperpolarization. However, how HCO3 − is transported into the sperm is not well understood. There is evidence that CFTR activity is involved in the human sperm capacitation but how this channel is integrated in the complex signaling cascades associated with this process remains largely unknown. In the present work, we have analyzed the extent to which CFTR regulates different events in human sperm capacitation. We observed that inhibition of CFTR affects HCO3 −-entrance dependent events resulting in lower PKA activity. CFTR inhibition also affected cAMP/PKA-downstream events such as the increase in tyrosine phosphorylation, hyperactivated motility, and acrosome reaction. In addition, we demonstrated for the first time, that CFTR and PKA activity are essential for the regulation of intracellular pH, and membrane potential in human sperm. Addition of permeable cAMP partially recovered all the PKA-dependent events altered in the presence of inh-172 which is consistent with a role of CFTR upstream of PKA activation.Fil: Puga Molina, Lis del Carmen. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Pinto, Nicolás Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Torres Rodríguez, Paulina. Universidad Nacional Autónoma de México; MéxicoFil: Romarowski, Ana. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Vicens Sanchez, Alberto. Universidad Nacional Autónoma de México; MéxicoFil: Visconti, Pablo E.. University of Massachussets; Estados UnidosFil: Darszon, Alberto. Universidad Nacional Autónoma de México; MéxicoFil: Treviño, Claudia L.. Universidad Nacional Autónoma de México; MéxicoFil: Buffone, Mariano Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentin

Pancreatic ductal deletion of Hnf1b disrupts exocrine homeostasis, leads to pancreatitis and facilitates tumorigenesis

BACKGROUND AND AIMS: The exocrine pancreas consists of acinar cells that produce digestive enzymes transported to the intestine through a branched ductal epithelium. Chronic pancreatitis is characterized by progressive inflammation, fibrosis and loss of acinar tissue. These changes of the exocrine tissue are risk factors for pancreatic cancer. The cause of chronic pancreatitis cannot be identified in one-quarter of patients. Here, we investigated how duct dysfunction could contribute to pancreatitis development. METHODS: The transcription factor Hnf1b, first expressed in pancreatic progenitors, is strictly restricted to ductal cells from late embryogenesis. We have previously shown that Hnf1b is crucial for pancreas morphogenesis but its postnatal role still remains unelucidated. To investigate the role of pancreatic ducts in exocrine homeostasis, we inactivated Hnf1b gene in vivo in mouse ductal cells. RESULTS: We uncovered that postnatal Hnf1b inactivation in pancreatic ducts leads to chronic pancreatitis in adults. Hnf1bΔduct mutants display dilatation of ducts, loss of acinar cells, acinar-to-ductal metaplasia (ADM) and lipomatosis. We deciphered the early events involved, with downregulation of cystic disease-associated genes, loss of primary cilia, upregulation of signaling pathways, especially Yap pathway involved in ADM. Remarkably, Hnf1bΔduct mutants developed pancreatic intraepithelial neoplasia and promote PanIN progression in concert with KRAS. We further showed that adult Hnf1b inactivation in pancreatic ducts is associated with impaired regeneration after injury, with persistent metaplasia and initiation of neoplasia. CONCLUSION: Loss of Hnf1b in ductal cells leads to chronic pancreatitis and neoplasia. This reveals that Hnf1b deficiency may contribute to diseases of the exocrine pancreas and could gain further insight into the etiology of pancreatitis and tumorigenesis.Support to CH was received from theCentre National de la Recherche Scientifique (CNRS), the Universite Pierre et Marie Curie (UPMC)- Sorbonne Université , the GEFLUC - Les entreprises contre le Cancer, the Societe Francophone du Diabete (SFD)-Ypsomed, the programme Emergence UPMC. EQ was supported by a PhD fellowship from the French Ministère de la Recherche et de la Technologie. MF is an assistant engineer of the CNRS. TD and AS were supported by Sorbonne Université. MDV was supported by a PhD student fellowship from the European Marie Curie Initial Training Network (ITN)-Biology of Liver and Pancreatic Development and Disease (BOLD). O. O. was supported by a Master1 fellowship. RCP was supported by a postdoctoral fellowship from the American Heart Association (14POST20380262). MG was supported by the National Institutes of Health (U01 DK089540) and the Juvenile Diabetes Research Foundation (1-2011-592). CH is a permanent senior researcher of the Institut National de la Sante et de la Recherche Medicale (INSERM).S

HSMA_WOA: A hybrid novel Slime mould algorithm with whale optimization algorithm for tackling the image segmentation problem of chest X-ray images

Recently, a novel virus called COVID-19 has pervasive worldwide, starting from China and moving to all the world to eliminate a lot of persons. Many attempts have been experimented to identify the infection with COVID-19. The X-ray images were one of the attempts to detect the influence of COVID-19 on the infected persons from involving those experiments. According to the X-ray analysis, bilateral pulmonary parenchymal ground-glass and consolidative pulmonary opacities can be caused by COVID-19 — sometimes with a rounded morphology and a peripheral lung distribution. But unfortunately, the specification or if the person infected with COVID-19 or not is so hard under the X-ray images. X-ray images could be classified using the machine learning techniques to specify if the person infected severely, mild, or not infected. To improve the classification accuracy of the machine learning, the region of interest within the image that contains the features of COVID-19 must be extracted. This problem is called the image segmentation problem (ISP). Many techniques have been proposed to overcome ISP. The most commonly used technique due to its simplicity, speed, and accuracy are threshold-based segmentation. This paper proposes a new hybrid approach based on the thresholding technique to overcome ISP for COVID-19 chest X-ray images by integrating a novel meta-heuristic algorithm known as a slime mold algorithm (SMA) with the whale optimization algorithm to maximize the Kapur's entropy. The performance of integrated SMA has been evaluated on 12 chest X-ray images with threshold levels up to 30 and compared with five algorithms: Lshade algorithm, whale optimization algorithm (WOA), FireFly algorithm (FFA), Harris-hawks algorithm (HHA), salp swarm algorithms (SSA), and the standard SMA. The experimental results demonstrate that the proposed algorithm outperforms SMA under Kapur's entropy for all the metrics used and the standard SMA could perform better than the other algorithms in the comparison under all the metrics

Morphological evidences indicate that the interference of cimetidine on the peritubular components is responsible for detachment and apoptosis of Sertoli cells

Cimetidine, referred as antiandrogenic agent, has caused alterations in the seminiferous tubules, including alterations in the peritubular tissue and death of myoid cells by apoptosis. Regarding the structural and functional importance of the peritubular tissue for the maintenance of Sertoli cells (SC), we purpose to investigate the SC-basement membrane interface, focusing the morphological features of SC and their interaction with the basement membrane in the affected tubules by cimetidine. Ten animals were distributed into two groups, control (CG) and cimetidine (CmG) which received saline solution and 50 mg of cimetidine per kg of body weight, respectively, for 52 days. The testes were fixed, dehydrated and embedded for analyses under light and transmission electron microscopy. Paraffin sections were submitted to the TUNEL method; sections of testes embedded in glycol methacrylate were submitted to PAS method and stained by H&E for morphological and quantitative analyses of Sertoli Cells. In the CmG, the SC nuclei were positive to the TUNEL method and showed typical morphological alterations of cell death by apoptosis (from early to advanced stages). A significant reduction in the number of Sertoli Cells was probably due to death of these cells by apoptosis. A close relationship between SC nuclear alterations (including a high frequency of dislocated nuclei from the basal portion) and damage in the peritubular tissue was observed. The ultrastructural analysis showed a parallelism between the gradual advancement of apoptotic process in SC and detachment of the anchoring sites (hemidesmosomes) of SC plasma membrane from the lamina densa. The presence of portions of lamina densa underlying the detached hemidesmosomes indicates a continuous deposition of lamina densa, resulting in the thickening of the basal lamina. The results indicate a possible disarrangement of the SC cytoskeleton, including the focal adhesion structure. These alterations are related to SC apoptosis and probably result from disturbs induced by cimetidine on the peritubular tissue

Congenital diaphragmatic hernia and retinoids: searching for an etiology

Congenital diaphragmatic hernia (CDH) is a major life-threatening cause of respiratory failure in the newborn. Recent data reveal the role of a retinoid-signaling pathway disruption in the pathogenesis of CDH. We describe the epidemiology and pathophysiology of human CDH, the metabolism of retinoids and the implications of retinoids in the development of the diaphragm and lung. Finally, we describe the existing evidence of a disruption of the retinoid-signaling pathway in CDH

Dynamics of notch pathway expression during mouse testis post-natal development and along the spermatogenic cycle

Articles in International JournalsThe transcription and expression patterns of Notch pathway components (Notch 1–3, Delta1 and 4, Jagged1) and effectors (Hes1, Hes2, Hes5 and Nrarp) were evaluated (through RT-PCR and IHC) in the mouse testis at key moments of post-natal development, and along the adult spermatogenic cycle. Notch pathway components and effectors are transcribed in the testis and expressed in germ, Sertoli and Leydig cells, and each Notch component shows a specific cell-type and timewindow expression pattern. This expression at key testis developmental events prompt for a role of Notch signaling in prepubertal spermatogonia quiescence, onset of spermatogenesis, and regulation of the spermatogenic cycle

Anion transporter TAT1 (SLC26A8) : physiological role and involvement in human asthenozoospermia

La protéine TAT1 (Testis Anion Transporter 1 ; SLC26A8) appartient à la famille des SLC26, une famille de transporteurs d’anions qui contribuent dans différents épithelia à l’homéostasie cellulaire. La protéine TAT1 s’exprime exclusivement dans les cellules germinales mâles, chez l’homme et chez la souris. Sur le spermatozoïde mature, la protéine TAT1 est localisée à la jonction des pièces intermédiaire (PI) et principale (PP) du flagelle, au niveau de l’annulus, une structure en forme d’anneau composée de différents polymères de Septines (1, 4, 6, 7 et 12).Le modèle murin d’invalidation du gène Tat1 présente une infertilité mâle par asthénozoospermie totale (absence de mobilité des spermatozoïdes) et des défauts de capacitation associés à des anomalies structurales du flagelle (plicature du flagelle, disjonction entre la PI et la PP, atrophie de l’annulus). Ce modèle indique que la protéine TAT1 pourrait avoir un rôle structural dans le maintien de l’annulus et dans la mise en place du flagelle. Par ailleurs, la protéine TAT1 possédant une activité de transport d’anions, il est vraisemblable qu’elle puisse influer directement sur la régulation de la mobilité et de la capacitation puisqu’il est bien établi que les échanges ioniques sont essentiels au contrôle de ces deux processus.En effet, les ions chlorure, bicarbonate et calcium participent à l’activation de la voie de signalisation AMPc/PKA, au cours des processus de mobilité et de capacitation (i.e. processus de maturation ayant lieu dans le tractus génital féminin et conférant au spermatozoïde un mouvement hyperactivé et la capacité à interagir avec l’ovocyte).Plusieurs travaux ont montré une interaction physique et fonctionnelle des membres de la famille SLC26 avec le canal chlorure/bicarbonate CFTR (Cystic Fibrosis Transmembrane conductance Regulator) dont les mutations sont responsables de la mucoviscidose. De manière intéressante des données récentes ont montré l’expression de CFTR dans le spermatozoïde et son rôle dans la régulation des flux de chlorure au cours de la capacitation. Au cours de ma thèse, nous avons testé la coopération entre les protéines TAT1 et CFTR ; nous avons pu montrer que la protéine TAT1 est capable d’interagir physiquement avec CFTR et de stimuler son activité de transport d’anions, suggérant qu’in vivo les deux protéines forment un complexe moléculaire impliqué dans la régulation des flux de chlorure et de bicarbonate dans le spermatozoïde.Tout comme TAT1, plusieurs membres de la famille SLC26 ont une expression tissulaire spécifique. Par ailleurs, les mutations génétiques de certains SLC26 sont associées à des pathologies humaines (surdité, diarrhée chlorurée congénitale et chondrodysplasie). De par le phénotype du modèle murin Tat1 et l’importance des SLC26 en pathologie humaine, TAT1 constitue un bon candidat dans la recherche des causes génétiques des asthénozoospermies humaines.Le laboratoire a mis en place au cours de ma thèse, un projet de recherche de mutations du gène TAT1 dans les asthénozoospermies humaines. Le séquençage des régions codantes du gène TAT1 dans une cohorte de 147 hommes infertiles par asthénozoospermie a ainsi permis d’identifier des variations de séquence inédites du gène chez 7 sujets. L’étude in vitro de certains variants indique pour trois d’entre eux une instabilité des formes mutantes associée à un défaut de stimulation du canal CFTR, in vitro. Par ailleurs, les spermatozoïdes de ces patients présentent d’importantes anomalies flagellaires dans la mise en place de la pièce intermédiaire, compatible avec un rôle de la protéine TAT1 et de ses partenaires (les septines) dans la genèse du flagelleTAT1 (Testis Anion Transporter 1 ; SLC26A8) belongs to the SLC26 family of anion transporters, which is implicated in cellular homeostasis of different epithelia. TAT1 is exclusively expressed in male germ cells, in human and mouse. On mature spermatozoa, TAT1 is located at the annulus, a ring-shaped structure composed of different septins polymers (1, 4, 6, 7 and 12), at the junction of the midpiece (MP) and principal piece (PP) of the flagellum.The knock-out mouse model of Tat1 gene shows a male infertility by complete asthenozoospermia (lack of sperm motility) and capacitation defects combined with flagellar structural abnormalities (flagella bending, MP and PP disjunction and atrophy of the annulus). This model suggests that the TAT1 protein could fulfill structural roles in the annulus and during flagellum biogenesis. Moreover TAT1 displayind an anion transport activity, it could also be implicated in the control of sperm motility and capacitation by regulating anions exchannges, which are well known to be essential for both processes.Indeed, chloride, bicarbonate and calcium ions are involved in the activation of the cAMP/PKA pathway, controlling sperm motility and capacitation processes (i.e. maturation events occuring in the female genital tract and providing the spermatozoa an hyperactivation movement and the ability to interact with oocyte).Several publications have reported a physical and functionnal interaction between SLC26 family members and the chloride/bicarbonate CFTR channel (Cystic Fibrosis Transmembrane conductance Regulator), which mutations are responsible of cystic fibrosis. Interestingly, recent data showed CFTR expression in spermatozoa and its role in the regulation of chloride fluxes during capacitation. During my thesis, we tested TAT1 and CFTR cooperation; we showed that TAT1 can interact physically with CFTR and stimulate its anion transport activity, suggesting that in vivo they form a molecular complex involved in the regulation of chloride and bicarbonate fluxes during sperm capacitation.Like TAT1, several SLC26 family members have a tissue specific expression. Furthermore genetic mutations in several SLC26 members result in human pathology such as deafness, congenital chloride diarrhea and chondrodysplasia. According to the phenotype of the KO Tat1 mouse model and the role of SLC26 members in human pathology, TAT1 constitutes a good candidate for the search of genetic causes of human asthenozoospermia.During my thesis, the laboratory has set up, a research project aiming at identifying mutations in the TAT1 gene that are responsible for human asthenozoospermia.Sequencing of the TAT1 gene coding regions in a cohort of 147 infertile men presenting with asthenozoospermia allowed us to identify several new sequence variations in in the TAT1 gene. In vitro study of these variants shows that 3 of them are associated with protein instability and abrogate CFTR stimulation. Besides, patients sperm show important flagellar abnormalities in the midpiece, consistent with a role of TAT1 and its partners (septins) in flagellum biogenesis