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In order to improve (re)production, the tissue specific expression profile of gene(s) involved in the various stages of reproduction should be studied thoroughly. For the same, Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) analysis of target genes in various biological samples could fulfil the aim. Various published literatures are evidence that the qRT-PCR technique is one of the most widely used and practical methods for detecting gene expression levels. In this technique, the use of housekeeping or reference gene acts as a calibrator, plays a vital role for quantification of mRNA expression levels of target gene in order to achieve objective and reliable findings. Housekeeping genes are responsible for the maintenance of basic cellular activities that are required for a cell's existence, regardless of the cell's specialised role in the tissue or organism. However, designing of primers and optimization of PCR conditions for a housekeeping gene was required to study the function of various other genes in buffalo endometrium. Therefore, the present study was aimed to optimize the β-actin gene as a housekeeping gene that stably expresses in the buffalo uterine endometrium, and establish β-actin gene primers for qRT-PCR technique.

Vitrification of preantral follicles is a promising technique to preserve female fertility. The aim of the present study was to evaluate the effect of supplementation of α-tocopherol in the vitrification solution on the viability, lipid peroxidation and mRNA expression of superoxide dismutases (SOD1 and SOD2) in vitrified cultured ovine preantral follicles at day-6 and day-12. Preantral follicles (200-300 µm) were isolated from the ovine ovaries by the mechanical method and were distributed separately to the vitrification medium supplemented with 10 mM and 20 mM of α-tocopherol. After a week, the follicles were thawed and analyzed for follicular viability by trypan blue dye exclusion method and subjected to in vitro culture (IVC) for 6 and 12 days. Our results revealed that the significant increase (p

Colorectal cancer (CRC) is the third most diagnosed cancer in the world. A better understanding of the molecular mechanism of CRC is essential for making novel strategies for the CRC management and its prevention. The present study aims to explore the molecular mechanism through integrated bioinformatics analysis by analyzing genes and their co-expression pattern in normal and CRC states. GSE110223, GSE110224 and GSE113513 gene expression profiles were analyzed in this study. The co-expression networks for normal and tumor samples were constructed separately and analyzed to identify the modules, sub-networks and key genes. Gene regulatory network analysis was done to understand the regulatory mechanism of selected genes. Survival analysis was performed for the identified sub-networks and key genes to understand their role in CRC progression. A total of seven modules were detected and the KEGG pathway analysis revealed these modules were mainly enriched with cell cycle, metabolism and signaling-related pathways. E2F6 and ETV4 transcription factors regulating the activity of multiple genes of identified modules were found to be up-regulated in CRC. Six Sub-networks and seven key genes, BORA, CCT7, DTL, RUVBL1, RUVBL2, THEM6 and TMEM97 associated with the CRC progression were identified. Disease-gene association analysis identified a novel association of the BORA gene with CRC that activates and regulates the AURORA-PLK1 cascades in the cell cycle. Survival analysis indicates that the overexpressed BORA is associated with unfavourable overall survival in CRC. The mechanistic role of BORA in the regulation of cell cycle progression suggests that BORA might act as a potential therapeutic target for CRC.

The trial was aimed at evaluating probable superiority, if any of nano zinc (NZn) over inorganic zinc (Zn) on immunity, serum minerals and T3 , T4 , and IGF-1 hormone profiles in goats. NZn was synthesized by using 0.45 M aqueous solution of Zn nitrate and 0.9 M aqueous solution of sodium hydroxide (average particle size 74 nm). Twenty-four male goats were grouped into four groups as per their body weight and were supplemented with either a basal diet with concentrate and straw at 50:50 ratio (Negative control, NC) alone or supplemented with 50 mg/kg Zn (Control) from inorganic Zn source, that is ZnO (IZn-50), 50 mg/kg Zn from NZn (NZn-50) or 25 mg/kg Zn from NZn (NZn-25). No change was observed in thyroid hormone status on zero and 90th day of experimental feeding, but NZn supplementation improved (p CD 4 + (cluster of differentiation in %) was more (p CD 8 + count than NC and similar (p > 0.05) to IZn-50 and NZn-25 groups without affecting (p > 0.05) the ratio of CD 4 + , CD 8 + among the treatment groups. Thus, NZn supplementation at 25 mg/kg had similar immunity and serum T3 , T4 and IGF-1 profiles compared with IZn at 50 mg/kg dose.

International audience; Background Quinacrine (QC) is popular for its anti-malarial activity. It has been reported exhibiting anti-cancerous properties by suppressing nuclear factor-κB and activating p53 signaling; however, its effect on cellular pathways in human non-small cell lung cancer (NSCLC) has not been studied. Materials and Methods Binding of QC with GSTA1 was studied computationally as well as through GST activity assay kit. Cell viability, cell cycle and mitochondrial membrane potential activity were studied using flow cytometry. RT-PCR and western blot were carried out to understand the involvement of various genes at their mRNA as well as protein level. Results QC inhibited the activity of GSTA1 approximately by 40-45% which inhibits cell survival and promotes apoptosis. QC reduced viability of NSCLC cells in a dose-dependent manner. It also causes nuclear fragmentation, G1/S arrest of cell cycle and ROS generation; which along with disruption of mitochondrial membrane potential activity leads to apoptotic fate. Conclusions Results revealed, QC has promising anti-cancer potential against NSCLC cells via inhibition of GSTA1, induction of G1/S arrest and ROS mediated apoptotic signaling. Copyright © Kumar et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License 3.0 (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Introduction: Lagenaria siceraria has various pharmacological effects like antioxidant, anti-hyperglycemic, anti-ulcer, anti-cancer, hepatoprotective, antimicrobial, anti-hyperlipidemic, anti-inflammatory and cardioprotective activities and is used to treat various diseases like diabetes, jaundice, piles, ulcers, hypertension, colitis, insanity, and skin diseases.Context and purpose of the study: The aim of the present study was to evaluate the phytochemical content, antioxidant activity, DNA damage and hepatoprotective activities of Lagenaria siceraria (LS)plant’s parts to explore their nutraceutical value.Methods: Phytochemical content was measured by the estimation of total phenolics, carotenoids, ascorbic acid, protein and carbohydrates while antioxidant activities were investigated by assaying the parameters of free radical scavenging activity, lipid peroxidation, superoxide anion radical scavenging activity, reducing power, and DNA damage protection assay. Hepatoprotective activity of LS leaf was measured by assaying cell viability, superoxide dismutase activity, malondialdehyde and nitric oxide formation.Results: Among the tested plant samples, LS leaves showed high contents of total phenolics, carotenoids, ascorbic acid, and protein contents whereas LS fruits have high carbohydrate contents. Evaluation of antioxidant activities in different parts of the LS plant showed that the LS leaf has high antioxidant properties as compared to other plant parts. The L. siceraria leaf also showed DNA protection activities against Fenton’s reagent as well as UV induced hydroxyl radical’s damage. To test the hepatoprotective activity of LS leaf against acetaminophen, induced hepatocellular toxicity and leaf extract concentrations of 2.5, 5.0 and 7.5 μg/104 cells were administered to cultured hepatocytes before acetaminophen exposure. The experimental results showed that LS leaf extract restores superoxide dismutase activity and reduces malondialdehyde and nitric oxide formation as compared to acetaminophen alone treated cultured cells.Conclusion: Our findings suggest that leaves of L. siceraria plants have high antioxidant potential and DNA damage-protective and hepatoprotective activities. Hence, the leaf part of this plant may be used in different food applications to provide nutritional and health benefits, as well as in various liver care herbal formulations.Keywords: Antioxidant, Hepatoprotective activity, Hepatocytes, Lipid peroxidation, Total phenolics

Background: Diabetes mellitus, is a multifactorial disease brought on by a complex interplay of metabolic, genetic, and lifestyle variables. Prolonged and chronic hyperglycemia is a complication of diabetes and might increase the risk of major health issues.Objective: This investigation aims to determine whether the phenolic phytochemical syringic acid (SA) has any protective role on the pancreas of diabetic rats.Methodology: Streptozotocin was injected intraperitoneally (40 mg/kg) into male Sprague-Dawley rats to induce diabetes. At a dosage of 50 mg per kg body weight, syringic acid (SA) was administered using an oral tube, once a day for 60 days. Our study examined plasma insulin, glucose, glycated hemoglobin, toxicity markers and antioxidant enzymes. The results were compared with those of diabetic rats receiving glimepiride (0.1 mg/kg) as the standard drug.Results: Treatment with syringic acid significantly lowered hyperglycemia, improved insulin levels, reduced toxicity markers in diabetic rats. Further, Syringic acid also promoted activity of enzymes such as catalase, glutathione peroxidase, glutathione reductase, and superoxide dismutase in the pancreas.Conclusion: These results imply that syringic acid, owing to its ability to control hyperglycemia, and reduce oxidative stress, affords antioxidant protection in the pancreas of diabetic rats.Keywords: Diabetes mellitus, Syringic acid, Antioxidant protection, Glimepiride.

Serine Proteinase Associated Disintegrin-1 (SPAD-1) is a low molecular mass (26 kDa) positively charged protein purified from Russell's viper venom (RVV) possessing cytotoxic activity on MCF7, human breast cancer cells. Primary sequence analysis of the protein confirms that it is a novel Snake Venom Serine Proteinase (SVSP) and a member of the trypsin family. SPAD-1 contains a conserved triad of Histidine (H), Aspartic acid(D) and Serine(S) residues at its active site for proteinase activity and also an adjacent histidine-glycine-aspartic acid (HGD) disintegrin-like motif. The serine proteinase and disintegrin parts are functionally active and independent. SPAD-1 showed proteolytic digestion of fibrinogen and fibronectin, but laminin digestion was below the detectable limit. Proteolytically inactivated SPAD-1 inhibited collagen and ADP-induced platelet aggregation. This study proposes considering Serine Proteinase Associated Disintegrin (SPAD) as a new group of snake venom proteins. Members of this group contain a serine proteinase catalytic triad and a disintegrin-like motif. SPAD-1 caused visible morphological changes in MCF7 cells, including a reduction of the cell-to-cell attachments, rounding of cell shape and death, in vitro. SPAD-1 also showed a dose-dependent significant decrease in the invasive potency of breast cancer cells. Confocal microscopic analysis revealed the breakage of nuclei of the SPAD-1-treated cells. SPAD-1 also increased cell detachment from the poly L-lysine-coated, laminin-coated and fibronectin-coated culture plate matrices, confirming the disintegrin activity. This study concludes that SPAD-1 may be a good candidate for anti-tumour drug design in the future.

Nutraceuticals derived from a medicinal plant having therapeutic significance are a gift to mankind to acquire healthy life. Carica papaya Linn. (aricaceae) commonly known as papaya. It is a significant fruit tree and is found in tropical and subtropical parts of the world. The extract from various parts of plant especially fruit and leaves contains many phytonutrients viz; vitamin A, B1, and vitamin C, calcium hydrate charcoal, phosphorus, iron, protein as well as some endopeptidases like namely papain, glycyl endopeptidase, chymopapain, and caricain. The extract of this plant is identified to be efficacious against diversified ailments like malaria, inflammation, digestive disorder, Microbes, Fungi, and many infectious disorders. Its prophylactic and therapeutic values enhance immunity and ensure a healthier life. Each part of this plant has its nutraceutical properties.Keywords: Nutraceuticals, Carica papaya, Papain, Caricain, Chymopapain

Background: Diabetes mellitus causes changes in the structural or functional anatomy of the heart. A high blood glucose level and oxidative stress are key factors in diabetic cardiac damage. Ferulic acid (4-hydroxy-3-methoxy cinnamic acid) is a biologically active compound in many functional foods like fruits, vegetables, and medicinal herbs. It belongs to the group of cinnamic acid derivatives.Objective: In the present study, we investigated the effects of Ferulic acid (FA) on Streptozotocin-induced diabetic cardiac damage in male Sprague Dawley rats.Materials and methods: A total of 30 male Sprague-Dawley rats were divided into five groups of six each. Diabetes was induced by a single intraperitoneal injection of Streptozotocin (STZ) (40 mg/kg body weight). Group I consisted of normal rats (N); group II consisted of normal rats treated with FA (N+FA); group III consisted of STZ-induced diabetic rats (D), and groups IV and V consisted of STZ-induced diabetic rats treated with FA at a dose of 50 mg/kg body weight and glibenclamide at a dose of 5 mg/kg body weight respectively (N+FA and N+G) for 60 days. Rats were sacrificed after the treatment period, and blood and heart tissue were collected for analysis.Results: STZ injection significantly increased blood glucose, HbA1c, cardiac marker enzymes LDH, CK-MB, and oxidative stress in heart tissue. The oral administration of FA to diabetic rats for 60 days significantly improved diabetic markers, oxidative stress, and cardiac markers.Conclusion: The present study indicated that FA affords cardioprotective effect in diabetic rats, and this protection may be due in part to the attenuation of oxidative stress.Keywords: ferulic acid, streptozotocin, diabetes, cardiac damage, oxidative stress