Ahmed Amin

Atresia is a process in ovarian follicles that is regulated by hormone-induced apoptosis. During atresia, granulosa cell (GC) apoptosis is a key mechanism orchestrated through diverse signaling pathways. Cocaine- and amphetamine-regulated transcript (CART) signaling within ovarian GCs has been demonstrated to play a key role in the regulation of follicular atresia in cattle, pigs, and sheep. The present work aimed to investigate the potential local regulatory role of CART in GC apoptosis-induced follicular atresia in buffalo, focusing on the modulation of the AKT/GSK3β/β-catenin signaling pathways, which are the intracellular signaling pathways involved in cell viability. Our findings revealed increased expression of CARTPT and BAX and decreased levels of AKT, β-catenin, and CYP19A1 genes in atretic follicles compared to healthy follicles. Subsequently, CART treatment in the presence of FSH inhibited the FSH-induced increase in GC viability by reducing estradiol production and increasing apoptosis. This change was accompanied by an increase in the gene expression levels of both CARTPT and BAX. At the protein level, treatment with CART in the presence of FSH negatively affected the activity of AKT, β-catenin, and LEF1, while the activity of GSK3β was enhanced. In conclusion, our study shows how CART negatively influences buffalo GC viability, underlying the modulation of the AKT/GSK3β/β-catenin pathway and promoting apoptosis—a key factor in follicular atresia.

INTRODUCTION: Endodontic treatment requires chemo-mechanical preparation of root canal to minimize the intracanal bacterial load. However, mechanical debridement results in smear layer formation containing infected debris requiring its removal before obturation. AIM OF THE STUDY: To assess the efficacy of Morinda Citrifolia juice (MCJ) as final irrigation protocol associated with manual dynamic agitation on smear layer removal by using scanning electron microscope. MATERIALS AND METHODS: Forty-five human mandibular permanent premolar with single canals were decoronated to a standard length of 15mm and instrumented using PTN rotary files till X3. Roots were then divided into 3 equal groups (n=15) according to the final irrigant used: Group I: 6% MCJ, Group II: 6% MCJ + 17% EDTA, Group III (control group): 2.5% NaOCl + 17% EDTA. Manual dynamic agitation (MDA) was used to activate the irrigants in the 3 groups. After that, the teeth were prepared for scanning electron microscopy evaluation, and smear layer removal was assessed.RESULTS: Along complete canal length, no significant difference was found between 6%MCJ+17% EDTA and 2.5% NaOCl + 17% EDTA (p=0.29). However, a significant difference was noted between 6%MCJ and 2.5% NaOCl + 17% EDTA (p=0.001). In all groups, the coronal third showed the best results (P = 0.02) and the apical third showed the worst with (P = 0.007.CONCLUSION: The use of 6%MCJ+17% EDTA associated with MDA could be an alternative to 2.5% NaOCl + 17% EDTA for smear layer removal, however, MCJ alone isn’t effective in smear layer removal.

Acylglycerophosphate acyltransferases (AGPATs) are the rate-limiting enzymes for the de novo pathway of triacylglycerols (TAG) synthesis. Although AGPATs have been extensively explored by evolution, expression and functional studies, little is known on functional characterization of how many members of the AGPAT family are involved in TAG synthesis and their impact on the cell proliferation and apoptosis. Here, 13 AGPAT genes in buffalo were identified, of which 12 AGPAT gene pairs were orthologous between buffalo and cattle. Comparative transcriptomic analysis and real-time quantitative reverse transcription PCR (qRT-PCR) further showed that both AGPAT1 and AGPAT6 were highly expressed in milk samples of buffalo and cattle during lactation. Knockdown of AGPAT1 or AGPAT6 significantly decreased the TAG content of buffalo mammary epithelial cells (BuMECs) and bovine mammary epithelial cells (BoMECs) by regulating lipogenic gene expression (p < 0.05). Knockdown of AGPAT1 or AGPAT6 inhibited proliferation and apoptosis of BuMECs through the expression of marker genes associated with the proliferation and apoptosis (p < 0.05). Our data confirmed that both AGPAT1 and AGPAT6 could regulate TAG synthesis and growth of mammary epithelial cells in buffalo. These findings will have important implications for understanding the role of the AGPAT gene in buffalo milk performance.