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The study was undertaken to examine the relative abundance (RA) of the major developmental important candidate genes in different grades of immature oocytes (A-grade, B-grade, C-grade and D-grade) and various stages of in vitro -produced embryos (2-cell, 4-cell, 8–16-cell, morula, and blastocyst) of buffalo using RT-qPCR. Results showed that the RA of GLUT1 , CX43 , HSP70.1 and GDF9 was significantly higher ( P < 0.05) in the A-grade of oocytes than the C-grade and D-grade but did not differ significantly from the B-grade of oocytes. Similarly, RA of BMP15 and Survivin were significantly higher ( P < 0.05) in A-grade than the other grades of oocytes, however, poly(A) polymerase expression was not significantly different ( P > 0.05) among the immature oocytes. The expression of GLUT1 was significantly higher ( P < 0.05) in the blastocysts, but the expression of CX43 ( P < 0.05; P > 0.05), HSP70.1 ( P < 0.05; P > 0.05) and GDF9 ( P > 0.05) was higher at the 2-cell stage than the other stages of embryos. Interestingly, the expression levels of poly(A) polymerase ( P < 0.05), BMP15 ( P < 0.05; P > 0.05) and Survivin ( P > 0.05) were higher at the 8–16-cell stage than the other stages of embryos. It is concluded that A-grade of immature oocytes has shown more mRNA abundance for the major developmental important genes; therefore A-grade oocytes may be considered as the most developmentally competent and suitable for handmade cloning research in buffalo.

The cheap and easy availability of the Kinnow peel waste has reported various applications due to presence of multifunctional groups. Therefore, in present study we explored its application to synthesize N-Benzylideneaniline and its derivatives based on Schiff base reaction. Kinnow peel powder is characterized by FTIR, TEM, SEM, XRD, EDX, and TGA for functional groups, morphology, surface, elements and thermal stability. Benzaldehyde, aniline, and their derivatives such as 4-methyl benzaldehyde, 4-hydroxy benzaldehyde, 4-methoxy benzaldehyde, and 4-methoxy aniline have been used to compare the efficacy of the Schiff base reaction using analysis of variance (ANOVA) and it has been observed that combination of Aniline and benzaldehyde for Schiff base reaction provided 85% yield of relative product.

The present study was undertaken to analyze the relative abundance (RA) of pluripotency-associated genes ( NANOG , OCT4 , SOX2 , c- MYC , and FOXD3 ) in different grades of immature oocytes and various stages of in vitro -produced buffalo embryos using RT-qPCR. Results showed that the RA of NANOG , OCT4 , and FOXD3 transcripts was significantly higher ( P < 0.05) in A grade oocytes compared with the other grades of oocytes. The RA of the c -MYC transcript was significantly higher ( P < 0.05) in A grade compared with the C and D grades of oocytes, but the values did not differ significantly from the B grade of oocytes. The RA of the SOX2 transcript was almost similar in all grades of the oocytes. The expression levels of NANOG ( P > 0.05), OCT4 ( P > 0.05), c- MYC ( P > 0.05) and SOX2 ( P < 0.05) were higher in the blastocysts compared with the other stages of the embryos. Markedly, FOXD3 expression was significantly higher ( P < 0.05) in 8–16-cell embryos compared with the 2-cell and 4-cell embryos and blastocyst, but did not differ significantly from the morula stage of the embryos. In the study, the majority of pluripotency-associated genes showed higher expression in A grade immature oocytes. Therefore, it is concluded that the A grade oocytes appeared to be more developmental competent and are suitable candidates for nuclear cloning research in buffalo. In buffalo, NANOG , OCT4 , SOX2 , and c- MYC are highly expressed in blastocysts compared with the other stages of embryos.

An Eco-catalysis approach reported in this manuscript, is developed by the kinnow peel waste as the green catalyst for the ring opening of epichlorohydrin with aromatic amines and its derivatives. The prepared green catalyst is used in three different variants i.e. kinnow peel powder (KPP), Bare component, and ash of KPP. Prepared catalyst characterized by TEM, SEM-EDX, FTIR, TGA, and XRD. Statistical analysis is a full factorial analysis of variance technique to identify the optimum combinations of types of catalyst, catalyst quantity, and derivative type.

Somatic cell nuclear transfer (SCNT) allows the multiplication of elite livestock and conservation of endangered species. However, restrictions on cow slaughter limit the access to oocytes for SCNT applications in Indian cattle breeds. To overcome this limitation, we utilized transvaginal ovum pick-up (OPU) method to collect oocytes, which were then used for the production of cloned embryos via the handmade cloning (HMC) technique. A total of 98 Sahiwal oocytes were collected, leading to the successful reconstruction of 24 SCNT Gir embryos. Out of these, five developed into blastocysts, which were transferred into five recipient cows. Two pregnancies were confirmed, but one was lost due to hydro-allantois condition. The other pregnancy continued to term, and a healthy Gir female calf weighing 32 kg was born. Microsatellite DNA analysis confirmed the genetic identity of the cloned calf to its donor. Postnatally, the calf was monitored for serum cytokine parameters, telomere length, and reproductive potentials. Cytokine profiling revealed variations between the cloned calf and naturally conceived counterparts; however, the born cloned calf did not exhibit any pathological conditions and has high telomere length compared to age-matched counterparts, and surviving well. Furthermore, to assess cloned cow utility in reproductive biotechnologies, we produced blastocyst stage embryos (35%) through OPU-IVF method and established one pregnancy from five transfers (20%). In conclusion, this study reports the first successful SCNT of Indian cattle breed and demonstrates the feasibility of the cloned cow for the production of OPU-IVF embryos.

In the present work, the Nickel oxide (rGO–NiO), Silver (rGO–Ag), Copper oxide (rGO–CuO) doped Graphene Oxide are reported for catalytic reactions. A comparative study for catalytic activities of these materials are performed with nitroaromatic compound 4-nitroaniline and the results are statistically studied by using univariate analysis of variance and Post Hoc Test through Statistical Package for Social Sciences and it is observed that CuO doped Graphene material is showing better catalytic activity in minimum time. So, further research has been focused on the catalytic acitivity of rGO–CuO only and it is found that it is efficient in reducing other nitro compounds also such as Picric acid and Nitrobenzene. Dye degradation of Methylene blue is also performed using CuO decorated Graphene material and significant changes were observed using UV spectroscopy. The characterization of rGO–CuO is done with Fourier-transform Infrared Spectroscopy, Powder X-ray Diffraction, Thermogravimetric Analysis, Scanning Electron Microscope and Transmission Electron Microscopy.

Silver nanoparticles (AgNPs) have been successfully synthesized using leaf extract of Neem ( Azadirachta Indica ), Mint ( Mentha Piperita ), Tulsi ( Ocimum Tenuiflorum ), Bermuda grass ( Cynodon Dactylon ) and silver salt. As plant extracts produce best capping material for the stabilization of nanoparticles. AgNPs were characterized by UV–Vis spectroscopy in range of 200–800 nm and transmission electron microscopy TEM, XRD and FTIR. The nanoparticles synthesized were mainly in sizes between 25 and 100 nm. They appeared to be spherical, nanotriangles and irregular in shape. Catalytic application was observed for all the aqueous solution of leaves, quantity taken was 1 ml, 2 ml, 3 ml, 4 ml and 5 ml. Furthermore, prepared Ag nanoparticles are also used for seed germination.

Garlic oils have promising possibilities for a wide range of applications in the food and pharmaceutical industries. Their widespread utilization is limited as they are lipophilic and highly volatile. Furthermore, they also possess strong odor and low physicochemical stability. Therefore, the present study aims to investigate the characteristics of garlic oil nano-emulsion through investigating its antifungal activity. The optimized nanoemulsion of D-Limonene and Garlic oil using 75% water and 20% emulsifiers having particle size range of 10 to 12 nm showed significant better antifungal activity against Tomato leaf spot disease without loss of antioxidant potential in comparison to garlic oil and D-Limonene as individual nanoemulsion. We have formulated Garlic oil, D-Limonene and Synergistic combination of both based nanoemulsion with antipest and antifungal properties. It is stabilized and particle size characterized by using Malvern Zeta sizer has been tested using and found the size in the range of 10.0 to 25.0 nm. It was further evaluated in field for its antifungal activity.

This research studies the effects of activation and inhibition of Wnt3A signaling pathway in buffalo (Bubalus bubalis) embryonic stem (ES) cell-like cells. To carry on this experimental study, the effects of activation and inhibition of Wnt3A signaling in buffalo ES cell-like cells were examined using Bio (0.5 mM) combined with WNT3A (200 ng/ml), as an activator, and Dickkopf-1 (Dkk1, 250 ng/ml), as an inhibitor, of the pathway. ES cells were cultured up to three weeks in ES cell medium without fibroblast growth factor-2 (FGF-2) and leukemia inhibitory factor (LIF), but in the presence of Bio, WNT3A, Bio+WNT3A and Dkk1. The effects of these supplements were measured on the mean area of ES cell colonies and on the expression levels of a number of important genes related to pluripotency (Oct4, Nanog, Sox2 and c-Myc) and the Wnt pathway (β-catenin). ES cell colonies cultured in ES cell medium that contained optimized quantities of LIF and FGF-2 were used as the control. Data were collected for week-1 and week-3 treated cultures. In addition, WNT3A-transfected ES cells were compared with the respective mock-transfected colonies, either alone or in combination with Dkk1 for expression of β-catenin and the pluripotency-related genes. Data were analyzed by ANOVA, and statistical significance was accepted at P<0.05. Among various examined concentrations of Bio (0.5-5 mM), the optimum effect was observed at the 0.5 mM dose as indicated by colony area and expressions of pluripotency-related genes at both weeks-1 and -3 culture periods. At this concentration,the expressions of Nanog, Oct3/4, Sox2, c-Myc and β-catenin genes were nonsignificantly higher compared to the controls. Expressions of these genes were highest in the Bio+WNT3A treated group, followed by the WNT3A and Bio-supplemented groups, and lowest in the Dkk1-treated group. The WNT-transfected colonies showed higher expressions compared to both mock and Dkk1-treated mock transfected colonies. WNT3A functions to maintain the pluripotency of ES cell-like cells both as an exogenous growth factor as well as an endogenously expressed gene. It complements the absence of FGF-2 and LIF, otherwise propounded essential for buffalo ES cell culture. WNT3A antagonizes the inhibitory effects of Dkk1 and acts in combination with its activator, Bio, to activate the Wnt signaling pathway.

In order to retain an undifferentiated pluripotent state, embryonic stem (ES) cells have to be cultured on feeder cell layers. However, use of feeder layers limits stem cell research, since experimental data may result from a combined ES cell and feeder cell response to various stimuli. In this experimental study, a buffalo ES cell line was established from in vitro derived blastocysts and characterized by the Alkaline phosphatase (AP) and immunoflourescence staining of various pluripotency markers. We examined the effect of various factors like fibroblast growth factor 2 (FGF-2), leukemia inhibitory factor (LIF) and Y-27632 to support the growth and maintenance of bubaline ES cells on gelatin coated dishes, in order to establish feeder free culture systems. We also analyzed the effect of feeder-conditioned media on stem cell growth in gelatin based cultures both in the presence as well as in the absence of the growth factors. The results showed that Y-27632, in the presence of FGF-2 and LIF, resulted in higher colony growth and increased expression of Nanog gene. Feeder-Conditioned Medium resulted in a significant increase in growth of buffalo ES cells on gelatin coated plates, however, feeder layer based cultures produced better results than gelatin based cultures. Feeder layers from buffalo fetal fibroblast cells can support buffalo ES cells for more than two years. We developed a feeder free culture system that can maintain buffalo ES cells in the short term, as well as feeder layer based culture that can support the long term maintenance of buffalo ES cells.