Analiza metilacije CpG kao moguće drugačije metode metilacije Xist gena u somatskih stanica koze (Capra hircus).

X chromosome inactivation is a complex epigenetic process that ensures equal expression of most Xlinked genes in both sexes. Xist (X-chromosome inactive specific transcript) is a crucial development related gene that initiates X chromosome inactivation in mammalian females during embryogenesis. DNA methylation is a major chromatin modification involved in transcriptional repression of genes, regulating X chromosome inactivation, genomic imprinting, and cell or tissue specific gene expression and in many other processes during embryo development. Appropriate DNA methylation of cytosine residues at CpG dinucleotide within the differential methylated region of many imprinted genes is required for normal post implantation development of a mammalian embryo. An aberrant pattern of DNA methylation because of incomplete or anomalous nuclear reprogramming of somatic cells is a reason for poor success rate of somatic cell nuclear transfer in farm animals. In the present study, a putative differential methylated region was identified for first time in goats (Capra hircus) followed by study of the DNA methylation status of this region, using bisulfite sequencing analysis taking fibroblast and cumulus cells from goats. The putative differential methylated region contains 10 CpG sites with a mean methylation level of 92% and 94% for fi broblast and cumulus cells respectively. Phylogenetic analysis of this region revealed goats and sheep positioned in a single cluster distinct from others studied. This study will be a definite help in future research on molecular events responsible for complete nuclear reprogramming of goat somatic cells in SCNT.Inaktivacija X kromosoma složeni je epigenski postupak kojim se osigurava jednaka ekspresija većine X-vezanih gena kod oba spola. Xist (X kromosom specifični inaktivirajući transkript) je ključni gen povezan s razvojem koji potiče inaktivaciju X kromosoma kod ženki sisavaca tijekom embriogeneze. Metilacija DNA glavna je modifikacija kromatina uključena u transkripcijsku regresiju gena, regulaciju X kromosomske inaktivacije, genomskog utiska, specifičnu staničnu ili tkivnu gensku ekspresiju odnosno u mnoge druge procese tijekom embrionalnog razvoja. Prikladna DNA metilacija citozin-rezidua na CpG dinukleotidu, unutar metilacijske regije mnogih utišanih gena, prijeko je potrebna za normalni postimplantacijski razvoj embrija sisavaca. Nepravilan oblik DNA metilacije zbog nekompletnog ili poremećenog reprogramiranja u jezgi somatskih stanica razlog je manje uspješnosti njezinog prijenosa kod farmskih životinja. U ovom radu istražena je moguća druga metilacijska regija koja je utvrđena prvi put u koze (Capra hircus). Navedeno je omogućeno nakon istraživanja statusa DNA metilacije u toj regiji, primjenom analize bisulfit sekvenciranja, uz upotrebu fibroblasta i stanice kumulusa koze. Moguća nova metilirajuća regija sadrži 10 CpG mjesta s prosječnom razinom metilacije od 92% i 94% za fibroblaste odnosno stanica kumulusa. Filogenetske analize regije pokazale su da su koze i ovce smještene u jedan odvojeni “grozd”. Istraživanje će pomoći u budućem radu vezanom za molekularnu razinu koja je odgovorna za potpuno reprogramiranje jezgre somatskih stanica koza u SCNT

Analiza metilacije CpG kao moguće drugačije metode metilacije Xist gena u somatskih stanica koze (Capra hircus).

X chromosome inactivation is a complex epigenetic process that ensures equal expression of most Xlinked genes in both sexes. Xist (X-chromosome inactive specific transcript) is a crucial development related gene that initiates X chromosome inactivation in mammalian females during embryogenesis. DNA methylation is a major chromatin modification involved in transcriptional repression of genes, regulating X chromosome inactivation, genomic imprinting, and cell or tissue specific gene expression and in many other processes during embryo development. Appropriate DNA methylation of cytosine residues at CpG dinucleotide within the differential methylated region of many imprinted genes is required for normal post implantation development of a mammalian embryo. An aberrant pattern of DNA methylation because of incomplete or anomalous nuclear reprogramming of somatic cells is a reason for poor success rate of somatic cell nuclear transfer in farm animals. In the present study, a putative differential methylated region was identified for first time in goats (Capra hircus) followed by study of the DNA methylation status of this region, using bisulfite sequencing analysis taking fibroblast and cumulus cells from goats. The putative differential methylated region contains 10 CpG sites with a mean methylation level of 92% and 94% for fi broblast and cumulus cells respectively. Phylogenetic analysis of this region revealed goats and sheep positioned in a single cluster distinct from others studied. This study will be a definite help in future research on molecular events responsible for complete nuclear reprogramming of goat somatic cells in SCNT.Inaktivacija X kromosoma složeni je epigenski postupak kojim se osigurava jednaka ekspresija većine X-vezanih gena kod oba spola. Xist (X kromosom specifični inaktivirajući transkript) je ključni gen povezan s razvojem koji potiče inaktivaciju X kromosoma kod ženki sisavaca tijekom embriogeneze. Metilacija DNA glavna je modifikacija kromatina uključena u transkripcijsku regresiju gena, regulaciju X kromosomske inaktivacije, genomskog utiska, specifičnu staničnu ili tkivnu gensku ekspresiju odnosno u mnoge druge procese tijekom embrionalnog razvoja. Prikladna DNA metilacija citozin-rezidua na CpG dinukleotidu, unutar metilacijske regije mnogih utišanih gena, prijeko je potrebna za normalni postimplantacijski razvoj embrija sisavaca. Nepravilan oblik DNA metilacije zbog nekompletnog ili poremećenog reprogramiranja u jezgi somatskih stanica razlog je manje uspješnosti njezinog prijenosa kod farmskih životinja. U ovom radu istražena je moguća druga metilacijska regija koja je utvrđena prvi put u koze (Capra hircus). Navedeno je omogućeno nakon istraživanja statusa DNA metilacije u toj regiji, primjenom analize bisulfit sekvenciranja, uz upotrebu fibroblasta i stanice kumulusa koze. Moguća nova metilirajuća regija sadrži 10 CpG mjesta s prosječnom razinom metilacije od 92% i 94% za fibroblaste odnosno stanica kumulusa. Filogenetske analize regije pokazale su da su koze i ovce smještene u jedan odvojeni “grozd”. Istraživanje će pomoći u budućem radu vezanom za molekularnu razinu koja je odgovorna za potpuno reprogramiranje jezgre somatskih stanica koza u SCNT

Caspases: An apoptosis mediator

The process of programmed cell death, or apoptosis, is generally characterized by distinct morphological characteristics and energy - dependent biochemical mechanisms. Apoptosis is a widely conserved phenomenon helping many processes, including normal cell turnover, proper development and functioning of the immune system, hormone dependent atrophy etc. Inappropriate apoptosis (either low level or high level) leads to many developmental abnormalities like, neurodegenerative diseases, ischemic damage, autoimmune disorders and many types of cancer. To use cells for therapeutic purposes through generating cell lines, it is critical to study the cell cycle machinery and signalling pathways that controls cell death and apoptosis. Apoptotic pathways provide a fundamental protective mechanism that decreases cellular sensitivity to damaging events and allow proper developmental process in multi-cellular organisms. Major mediator of apoptosis is a family of proteins known as caspases. There are mainly fourteen types of caspases but out of them only ten caspasese have got essential role in controlling the process of apoptosis. These ten caspases have been categorized into either initiator caspases (caspase 2, 8, 9, 10) or executioner caspases (caspase 3, 6, 7). Although various types of caspases have been identified so far, the exact mechanisms of action of these groups of proteins is still to be fully understood. The aim of this review is to provide a detail overview of role of different caspases in regulating the process of apoptosis

Complications of Thyroid Surgery - Our Experience

Abstract During the 1800s, the mortality rate from thyroid surgery was approximately 40%. Sterile surgical arenas , general anaesthesia, and improved surgical techniques have made death from thyroid surgery extremely rare today. By developing a thorough knowledge of the anatomy and of the ways to prevent each complication. The surgeon can minimize each patient's mortality & morbidity

Prolificacy in Raighar goats is independent of FecB gene

Aim: The Research was undertaken to find the association between FecB and high prolificacy in Raighar goats. Materials and Methods: DNA was extracted from blood, collected from does (n=101) with history of high prolificacy. Further tetra-primer amplification refractory mutation system (T-ARMS) PCR and agarose gel electrophoresis were followed to screen the mutation. Results: Raighar goats were found to be wild homozygotes suggesting absence of FecB mutation. Conclusion: Prolificacy in case of Raighar goats is not due to the mutation at FecB locus. It is thought to search for other genes or loci in goat fecundity. [Vet World 2013; 6(8.000): 479-481