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This study aimed to evaluate the effect of melatonin and melatonin-loaded chitosan nanoparticle (CMN) supplementation to maturation media on buffalo oocyte maturation rate and relative expression of genes: growth differentiation factor 9 (GDF9), bone morphogenetic protein 15 (BMP15), B Cell Lymphoma 2 (BCL2), Associated X protein (BAX) and superoxide dismutase 1 (SOD1). Buffalo ovaries were heaved from Al-Mounib slaughterhouse, cumulus-oocyte complexes (COCs) were in vitro matured in three different media, TCM-199 medium (control), TCM-199 with melatonin 10-9M, and TCM-199 with CMN 10-9M. The assessment of the nuclear maturation rate was carried out through the presence of the first polar body. In addition, the mature buffalo oocytes were stored on RNA later for genetic analysis of GDF9, BMP15, SOD1, BCL2, and BAX genes using quantitative real-time PCR (qRT-PCR). The results were reported that buffalo oocytes supplemented with melatonin-loaded chitosan nanoparticle (CMN) or melatonin have a significant effect on nuclear maturation rate 94.04±0.65 and 88.74±0.77 respectively when compared with buffalo oocytes matured with basic media (control) 79.67±1.35. Furthermore, buffalo oocytes supplemented with melatonin-loaded chitosan nanoparticle (CMN) or melatonin showed significant upregulation of GDF9, BMP15, SOD1, and BCL2 genes and significant downregulation of BAX gene when compared with oocyte matured with basic media (control). In conclusion, the results of nuclear maturation rate and relative expression pattern of GDF9, BMP15, SOD1, BCL2, and BAX reflect that melatonin-loaded chitosan nanoparticle (CMN) and melatonin' may play an important role in the buffalo oocytes developmental competence. [ABSTRACT FROM AUTHOR]

This study aimed to investigate the impact of zinc sulfate and nano zinc oxide on the In-vitro maturation (IVM) of oocytes and on the In-vitro embryo developmental competence of buffaloes. Ovaries were obtained from the abattoir. Good quality oocytes (excellent & good) were matured in tissue culture medium -199 (TCM-199) vs. TCM-199 +10-6M zinc sulfate vs. TCM-199 +10-6 M nano zinc oxide enriched by fetal calf serum 10% (FCS) + 10 μg/ml follicle-stimulating hormone + 50 μg/ml gentamicin. The oocyte maturation was done in the incubator for 22 h in a humidified environment with CO2 5% and 38.5°C. Frozen-thawed semen was used to fertilize Mature oocytes, which were then incubated for 18 hours, before being cultured by synthetic oviduct fluid (SOF) for 7 days. The obtained results showed that supplementing maturation medium with 10-6 M zinc sulfate and 10-6 M nano zinc oxide resulted in a significant (P<0.05) rise in GIII cumulus cell expansion of buffalo oocytes by 52.93 %, and 59.75%, respectively, as compared to oocytes cultured in free medium (36.8%). G0 cumulus cell expansion showed a significant (P<0.05) decrease in zinc sulfate and nano zinc oxide groups (7.85, 3.29 %, respectively) when compared with oocytes cultured in free medium (14.73 %). The rate of maturation of oocytes with polar bodies was significantly greater in the zinc sulfate and nano zinc oxide groups (86.98, and 92.43%, respectively) when compared with those matured in free medium (80.11%). The hatching (cleavage) rate was significantly greater (P<0.05) in the zinc sulfate and nano zinc oxide groups (83.17, 87.66 respectively %) when compared with the TCM-199 (free medium) group (78.60%). The transferable embryos (blastocyst & morula) rates significantly raised (P<0.05) in the zinc sulfate (17.28 & 19.87 %, respectively) and nano zinc oxide groups (21.23 and 26.21%, respectively) when compared with TCM-199 group (11.19 &13.75 %, respectively). In conclusion, in vitro maturation rate and transferable embryo rates in buffaloes improve by adding zinc sulfate and nano zinc oxide to the medium of maturation. [ABSTRACT FROM AUTHOR]

Background: Patients on continuous flow ventricular assist devices (CF‐VADs) are at high risk for the development of Acquired von‐Willebrand Syndrome (AVWS) and non‐surgical bleeding. von Willebrand Factor (vWF) plays an essential role in maintaining hemostasis via platelet binding to the damaged endothelium to facilitate coagulation. In CF‐VAD patients, degradation of vWF into low MW multimers that are inefficient in facilitating coagulation occurs and has been primarily attributed to the supraphysiological shear stress associated with the CF‐VAD impeller. Methods: In this review, we evaluate information from the literature regarding the unraveling behavior of surface‐immobilized vWF under pulsatile and continuous flow pertaining to: (A) the process of arterial endothelial vWF production and release into circulation, (B) the critical shear stress required to unravel surface bound versus soluble vWF which leads to degradation, and (C) the role of pulsatility in on the production and degradation of vWF. Results and Conclusion: Taken together, these data suggests that the loss of pulsatility and its impact on arterial endothelial cells plays an important role in the production, release, unraveling, and proteolytic degradation of vWF into low MW multimers, contributing to the development of AVWS. Restoration of pulsatility can potentially mitigate this issue by preventing AVWS and minimizing the risk of non‐surgical bleeding. [ABSTRACT FROM AUTHOR]

Background: Patients with continuous flow ventricular assist devices (CF‐VADs) are at high risk for non‐surgical bleeding, speculated to associate with the loss of pulsatility following CF‐VAD placement. It has been hypothesized that continuous shear stress causes elongation and increased enzymatic degradation of von Willebrand Factor (vWF), a key player in thrombus formation at sites of vascular damage. However, the role of loss of pulsatility on the unravelling behavior of vWF has not been widely explored. Methods: vWF molecules were immobilized on the surface of microfluidic devices and subjected to various pulsatile flow profiles, including continuous flow and pulsatile flow of different magnitudes, dQ/dt (i.e., first derivative of flow rate) of pulsatility and pulse frequencies to mimic in vivo shear flow environments with and without CF‐VAD support. VWF elongation was observed using total internal reflection fluorescence (TIRF) microscopy. Besides, the vWF level is measured from the patients' blood sample before and after CF‐VAD implantation from a clinical perspective. To our knowledge, this work is the first in providing direct, visual observation of single vWF molecule extension under controlled‐pulsatile shear flow. Results: Unravelling of vWF (total sample size n ~ 200 molecules) is significantly reduced under pulsatile flow (p < 0.01) compared to continuous flow. An increase in the magnitude of pulsatility further reduces unravelling lengths, while lower frequency of pulsatility (20 vs. 60 pulses per min) does not have a major effect on the maximum or minimum unravelling lengths. Evaluation of CF‐VAD patient blood samples (n = 13) demonstrates that vWF levels decreased by ~40% following CF‐VAD placement (p < 0.01), which correlates to single‐molecule observations from a clinical point of view. Conclusions: Pulsatile flow reduces unfolding of vWF compared to continuous flow and a lower pulse frequency of 20 pulses/minute yielded comparable vWF unfolding to 60 pulses/minute. These findings could shed light on non‐surgical bleeding associated with the loss of pulsatility following CF‐VAD placement. [ABSTRACT FROM AUTHOR]

Natural products play a pivotal role in innovative drug discovery by providing structural leads for the development of new therapeutic agents against various diseases.The present study aims to focus on the in silico assessment of the therapeutic potential of antidiabetic phytoconstituents which were identified and isolated from the extracts of Bauhinia rufescens Lam, a medicinal plant traditionally used for various pharmacotherapeutic purposes. The physicochemical and pharmacokinetic parameters of the previously isolated thirty eight compounds were predicted using SwissADME web tool whereas OSIRIS Property Explorer was used for toxicity risk assessment and drug- likeliness. Twelve compounds were selected for docking on human α-glucosidase and α-amylaseenzymes using Autodock 4.0 software. Furthermore, the active extract was in vivo tested for the antidiabetic activity and then identified usingTLC bioautographic method. Eriodictyol was found to have the highest potential as an inhibitor against α-amylase with binding energy of −9.92 kcal/mol. Rutin was the most potent against α-glucosidase with binding energy of-9.15 kcal/mol. A considerable number of hydrogen bonds and hydrophobic interactions were computed between the compounds and the enzymes thereby making them energetically favorable and suggesting inhibition of these two enzymes as a plausible molecular mechanism for their antidiabetic effect. These two flavonoids could therefore be used as potential leads for structure- based design of new effective hypoglycemic agents. [ABSTRACT FROM AUTHOR]

During mammalian preimplantation development, the transition from morula to blastocyst is a critical biological event. This process involves polarization and initial specification of lineages, regulated by various transcription factors that have been extensively studied in mice. Our single-cell RNA sequencing analyses revealed that TEAD3 is specifically expressed in the trophectoderm cells of bovine preimplantation embryos, unlike in mice. The objective of this study is to determine the functional role of TEAD3 in bovine preimplantation development. While TEAD3 knockdown does not affect blastocyst formation in cattle, embryos fail to progress to the blastocyst stage when both TEAD3 and TEAD4, another member of the TEAD family, are disrupted using RNA interference and base editing techniques, respectively. This finding suggests a redundant role for TEAD3 and TEAD4 in preimplantation development in cattle. RNA sequencing analysis identified dysregulation of 215 genes, with 53 genes upregulated and 162 genes downregulated. Notably, we observed a reduction in the expression of trophectoderm-specifier genes KRT8, KRT18, and EZR, as well as HIPPO signaling pathway components. Immunofluorescence analysis further revealed that the protein expression levels of KRT8 and EZR were significantly decreased. Importantly, the initial expression of trophectoderm lineage-specific factors such as TFAP2C and GATA3, as well as the inner cell mass lineage-specific transcription factor OCT4, remained unaffected. This contrasts with the role of TEAD4 in directly regulating trophectoderm lineage specification in mice. Thus, our studies demonstrate that TEAD3 and TEAD4 play essential and redundant roles upstream of TE fate decisions during preimplantation development in cattle.