Inhibition of NF-κB promotes autophagy via JNK signaling pathway in porcine granulosa cells.

The transcription factor nuclear factor-κB (NF-κB) plays an important role in diverse processes, including cell proliferation and differentiation, apoptosis and inflammation. However, the role of NF-κB in porcine follicle development is not clearly elucidated. In this study, we demonstrated that follicle stimulating hormone (FSH) increased the level of inhibitor of NF-κB (IκB) protein and promoted the cytoplasmic localization of p65, indicating that FSH inhibits the activation of NF-κB in porcine granulosa cells. Moreover, inhibition of NF-κB by FSH or another specific inhibitor of NF-κB, pyrrolidine dithiocarbamate (PDTC), could activate JNK signaling and enhance autophagic activity in porcine granulosa cells. Knockdown of RelA (p65) Subunit of NF-κB by RNA interference abrogated the activation of JNK signaling pathway and the increase of autophagic protein expression by FSH. Meanwhile, the functional significance of FSH or PDTC-mediated autophagy were further investigated. Our results demonstrated that the increased autophagy promoted progesterone secretion in porcine granulosa cells. Blockage of autophagy by chloroquine obviated the FSH or PDTC-induced progesterone production. Taken together, these results indicate that inhibition of NF-κB increased autophagy via JNK signaling, and promote steroidogenesis in porcine granulosa cells. Our results provide new insights into the regulation and function of autophagy in mammalian follicle development.

Recombinant bovine interferon-τ enhances in vitro development of bovine embryos by upregulating expression of connexin 43 and E-cadherin.

Interferon-τ (IFNT), produced in ruminants by embryonic trophoblastic cells before implantation, is involved in the maternal recognition of pregnancy. It is a pleiotropic molecule that alters the synthesis of endometrial proteins and inhibits the proliferation of some cells. The present study investigated the effects of recombinant bovine IFNT on the development of early-stage bovine embryos and the molecular mechanism underlying this effect. This study demonstrated that expression of mRNA encoding type I IFN receptor subunits was detectable from d 4 to 8 in in vitro fertilized (IVF) bovine embryos. A considerable number of IVF (n = 1,941) and parthenogenetic activated (n = 1,552) bovine embryos demonstrated that supplementing the culture medium with IFNT (100 ng/mL) produced a greater percentage of blastocysts, and the total cell number within the resulting blastocysts was higher. In addition, IFNT upregulated the expression levels of both mRNA and protein for connexin 43 (GJA1) and E-cadherin (CDH1) and expression levels for granulocyte-macrophage colony-stimulating factor and insulin-like growth factor 2 mRNA but not for their proteins in d 8 embryos. However, IFNT inhibited mRNA expression for leukemia inhibitory factor (LIF), LIF receptor α, and the sodium/potassium-transporting ATPase subunit ß-1. We concluded that IFNT promoted the development of bovine embryos by upregulating the expression of GJA1 and CDH1. Thus, supplementing embryo cultures or transfer medium with IFNT may stimulate embryo development and improve embryo transfer efficiency.

Investigating the impact of sous vide cooking on the eating quality of spent buffalo (BUBALUS BUBALIS) meat.

This study describes the impact of sous vide cooking at different temperatures and time intervals on the eating quality, specifically tenderness of two muscles, bicep femoris (BF) and semitendinosus (ST) from spent buffalo (Bubalus bubalis). Spent buffalo refers to water buffalo that are no longer considered productive following a sixth lactation cycle. Steaks from each muscle were obtained and cooked at three combinations of time and temperature, namely 55 °C-8H, 65 °C-5H, and 95 °C-45 M, respectively. Warner-Bratzler Shear Force (WBSF), cooking loss, cooking yield, color, water activity (aw), total water content (TWC), total collagen content (TCC), heat soluble collagen (HSC), myofibrillar fragmentation index (MFI), and sensory evaluation were measured. The collagen solubilization results showed that temperature and time interacted (P ≤ 0.05), reducing the toughness of the muscles. The tenderization achieved through sous vide cooking was mainly attributed to the thermal denaturation of proteins at the typically lower temperatures and extended time used, weakening of connective tissue through collagen solubilization, and water retention. More cooking loss (P ≤ 0.05) was observed at high temperature treatment of 95 °C-45 M. Meat color, TWC, MFI, and overall acceptability exhibited differences among treatments (P ≤ 0.05). An extended heat interval at lower temperatures caused initial denaturation of myofibrillar proteins, then solubilization of connective tissue proteins. Cooking treatment 55 °C-8H (P ≤ 0.05) reduced the WBSF in both muscles; however, the ST appeared more tender than BF.

In-vitro and in-vivo assessment of the anti-diabetic, analgesic, and anti-inflammatory potenstials of metal-based carboxylates derivative.

In the current research work, an amide based metal carboxylate chemical ([((5-((5-(2-hydroxyethyl)-4-methylthiazol-3-ium-3-yl)methyl)-2-methylpyrimidin-4-yl)amino)bis((4-((4-methoxy-2-nitrophenyl)amino)-4-oxobutanoyl)oxy)zinc]) was identified as anti-diabetic analgesic and anti-inflammatory. The identified chemical(MT-1) was tested for acute toxicity (the MT-1 was fund safe), antidiabetic analgesic, and anti-inflammatory potentials. The in-vitro study was conducted for antidiabetic enzyme inhibition (α-amylase and α-glucosidase) and the in-vivo studies included analgesic (acetic acid-induced writing and hot plate model) and anti-inflammatory (carrageenan etc induced edema) effects. The tested compound showed 88.63% (IC50 = 3.23 µg/ml) and 89.10%(IC50 = 5.10 µg/ml) againstα-amylase and α-glucosidase respectively. A significant (p < 0.001) analgesic effect was noted by MT-1 in acetic acid-induced animal models with a percent effect of 86.00, 60.,06, and 55.29 at the tested doses of 20, 1,0, and 5 mg/kg respectively. In the case of the hot plate model, the MT-1 showed a significant (p < 0.001) effect with maximum percent prolongation in latency observed after 60 min.08, 22.2,9, and 11.61) against 20, 1,0, and 5 mg/kg. The analgesic effect in the hot plate model was significantly (p < 0.01) reversed by the injection of naloxone (0.125 mg/kg). The paw edema induced by carrageenan, histamine, bradykinin, arachidonic acid, and PGE2 was significantly antagonized with percent attenuation of 34.09, 33.57, 34.60, 34.14, and 48.04 respectively. Furthermore, to predict the interactions between the MT-1 compound and COX-2 molecular docking was carried out and the result was compared with the standard compound. The docking score of MT-1 was predicted as -6.30 while that of Diclofenac was predicted as -6.82. Both compounds made several hydrogen bond interactions with the active site of the COX-2 enzyme. The docking study revealed the potent inhibitory potential of the compound MT-1 against the COX-2 receptor.

Expression of interferon-stimulated gene ISG15 and ubiquitination enzymes is upregulated in peripheral blood monocyte during early pregnancy in dairy cattle.

Pregnancy recognition is associated with the upregulation of the great number of genes in ruminants. One of these genes is interferon-stimulatory gene (ISG15), an ubiquitin-like protein that mediates the conjugation of different proteins through its ISGylation enzymes UBE1L and UBCH8. We demonstrate that ISG15 and its ISGlyation enzymes UBE1L and UBCH8 are differentially regulated in pregnant and non-pregnant cows. The mRNA and/or protein levels of ISG15, UBE1L and UBCH8 increased significantly (p<0.05) after treatment of PBMCs with different concentrations of interferon-tau (IFN-τ). To correlate these changes with pregnancy, cows were synchronized and artificially inseminated on day 0 (AI group). Cows that were not artificially inseminated served at the control (NON-AI group). Blood was collected from cows of both groups on day 18, and the mRNA levels of ISG15, UBE1L and UBCH8 in PBMCs were analyzed and compared between groups. Pregnancy was diagnosed on day 45 by rectal palpation. Compared with non-pregnant cows and the NON-AI group, the mRNA levels of ISG15, UBE1L and UBCH8 increased significantly (p<0.05) in pregnant cows. In conclusion, the secretion of IFN-τ by the conceptus acts on PBMCs to upregulate the interferon-stimulated gene ISG15, thereby activating UBE1L and UBCH8. Thus ISG15, UBE1l and UBCH8 genes are significantly upregulated in the AI- pregnant cows.