Important genes affecting fibre production in animals: A review

The realignment of the production profile to respond to demanding market signals is one of the most important challenges that an animal breeders face today. Animal fibre being a significant contributor to the agricultural economy needs special attention. This is especially true for sheep and goats where fibre production can account for as much as 20% of the total gross income. It is therefore necessary to gain a better insight into the genes governing wool traits. Gene mapping studies have identified some chromosomal regions influencing fibre quality and production. These may help in the selection of animals producing better quality wool. These are more efficient and accurate than the conventional techniques. This paper critically reviews various genes governing fibre growth in animals and their importance. Fibre quality and production genes may provide novel insights into our understanding of the science of genetics and breeding. The discovery of new fibre-related genes and their functions may also help in future studies related to fibre development and in the development of new and advanced techniques for the improvement of fibre production and quality

Scavenger receptor B1 facilitates the endocytosis of \u3ci\u3eEscherichia coli\u3c/i\u3e via TLR4 signaling in mammary gland infection

SCARB1 belongs to class B of Scavenger receptors (SRs) that are known to be involved in binding and endocytosis of various pathogens. SRs have emerging role in regulating innate immunity and host–pathogen interactions by acting in co-ordination with Toll-like receptors.Query Little is known about the function of SCARB1 in milk-derived mammary epithelial cells (MECs). This study reports the role of SCARB1 in infection and its potential association in TLR4 signaling on bacterial challenge in Goat mammary epithelial cells (GMECs). The novelty in the establishment of MEC culture lies in the method that aims to enhance the viability of the cells with intact characteristics upto a higher passage number. We represent MEC culture to be used as a potential infection model for deeper understanding of animal physiology especially around the mammary gland. On E.coli challenge the expression of SCARB1 was signifcant in induced GMECs at 6 h. Endoribonuclease-esiRNA based silencing of SCARB1 affects the expression of TLR4 and its pathways i.e. MyD88 and TRIF pathways on infection. Knockdown also affected the endocytosis of E.coli in GMECs demonstrating that E.coli uses SCARB1 function to gain entry in cells. Furthermore, we predict 3 unique protein structures of uncharacterized SCARB1 (Capra hircus) protein. Overall, we highlight SCARB1 as a main participant in host defence and its function in antibacterial advances to check mammary gland infections

IGF-1 Gene Polymorphism and its Association with Cashmere Fiber Trait in Changthangi Goats

The Cashmere (Pashmina) fiber attracts great demand worldwide for its fineness. To identify and determine the role of Insulin-like Growth Factor-1 (IGF-1) gene polymorphism on Pashmina fiber traits (quality and quantity), exon IV, and part of intron IV of the IGF-1 gene of Changthangi goats were analyzed. 103 Changthangi goats of either sex were divided into two groups, the high producers and low producers of Pashmina. Upon restriction digestion with HaeIII, three genotypes were identified, i.e., A1A1 (0.253), A2A2 (0.320), and A1A2 (0.427). These genotypes did not vary significantly (p > .05) from Hardy–Weinberg Equilibrium. DNA sequence analysis revealed G > C transversion in intron-IV of IGF-1 gene at the HaeIII restriction site. The SNP effect of two alleles on mean Cashmere fiber diameter of down fibers (MFD) was statistically significant (p < .05) and the average finest fiber diameter was 13.46 µ for genotype A2A2. The mean genetic variability parameters for the gene analyzed were heterozygosity value (0.4977), PIC Value (0.3738), Effective allele no (1.9908),and Shannon index I (0.6908). The significant association of IGF-1 gene polymorphism and fiber finesse can be exploited in breeding programmes to improve the production and quality of Pashmina fiber in goat populations

A Potent Inhibitor of Phosphoinositide 3-Kinase (PI3K) and Mitogen Activated Protein (MAP) Kinase Signalling, Quercetin (3, 3', 4', 5, 7-Pentahydroxyflavone) Promotes Cell Death in Ultraviolet (UV)-B-Irradiated B16F10 Melanoma Cells.

Ultraviolet (UV) radiation-induced skin damage contributes strongly to the formation of melanoma, a highly lethal form of skin cancer. Quercetin (Qu), the most widely consumed dietary bioflavonoid and well known inhibitor of phosphoinositide 3-kinase (PI3K) and mitogen activated protein (MAP) kinase signalling, has been reported to be chemopreventive in several forms of non-melanoma skin cancers. Here, we report that the treatment of ultraviolet (UV)-B-irradiated B16F10 melanoma cells with quercetin resulted in a dose dependent reduction in cell viability and increased apoptosis. The present study has brought out that the pro-apoptotic effects of quercetin in UVB-irradiated B16F10 cells are mediated through the elevation of intracellular reactive oxygen species (ROS) formation, calcium homeostasis imbalance, modulation of anti-oxidant defence response and depolarization of mitochondrial membrane potential (ΔΨM). Promotion of UVB-induced cell death by quercetin was further revealed by cleavage of chromosomal DNA, caspase activation, poly (ADP) ribose polymerase (PARP) cleavage, and an increase in sub-G1 cells. Quercetin markedly attenuated MEK-ERK signalling, influenced PI3K/Akt pathway, and potentially enhanced the UVB-induced NF-κB nuclear translocation. Furthermore, combined UVB and quercetin treatment decreased the ratio of Bcl-2 to that of Bax, and upregulated the expression of Bim and apoptosis inducing factor (AIF). Overall, these results suggest the possibility of using quercetin in combination with UVB as a possible treatment option for melanoma in future

Open pulled straw vitrification of in vitro matured sheep oocytes using different cryoprotectants

The aim of this study was to compare the survival and in vitro development of sheep oocytes after open pulled straw vitrification and different final concentrations of permeable cryoprotectants.In 5 identical replicates of two experiments, in vitro matured (IVM) oocytes were vitrified by the Open Pulled Straw (OPS) method, then warmed, and the surviving ones were subjected to parthenogenetic activation. In Experiment 1, survival rate of oocytes aftervitrification in 33% ethylene glycol was higher than in 33% DMSO or a mixture of 17.5% ethylene glycol and 17.5% DMSO (87.64 vs. 77.43 vs. 69.39%, respectively). The cleavage and blastocyst rates were higher after vitrification in mixture group than in ethylene glycol and DMSO (46.81 and 15.5 vs. 37.55 and 9.12 vs. 29.51 and 6.40%, for cleavage and blastocyst rates in different groups, respectively). In Experiment 2, elevated concentrations of vitrification solutions were used. The survival rate was higher after vitrification in 40% ethylene glycol and in the mixture of 20% ethylene glycol and 20% DMSO than in 40% DMSO(90.22 vs. 87.56 vs. 75.34%, respectively). Cleavage and blastocyst rates were also higher in the ethylene glycol and ethylene glycol – DMSO mixture group than in DMSO alone group (50.67 and17.60 vs. 49.13 and 14.45 vs. 33.86 and 9.81% for cleavage and blastocyst ratesin different groups, respectively). The survival rates between the two experimental groups was higher in 40% ethylene glycol group, 40% mixture group and 33% ethylene glycol group than in 40% DMSO group, 33% mixture group and 33% DMSO group. Cleavage and blastocystrates were higher in 40% ethylene glycol group, 40% mixture group and 33% mixture group than in 40% DMSO group, 33% ethylene glycol group and 33% DMSO group. All cleavage and blastocyst rates in both the experiments were lower than those of the non-vitrified controlgroup (87.00 and 45.00, respectively). In conclusion, although ethylene glycol group and ethylene glycol – DMSO mixture group gave better survival and cleavage – blastocyst rates than DMSO group, the survival rates were lower than the control group and hence the technique could be further improved to get better results after OPS vitrification of IVMsheep oocytes

Quercetin attenuates PI3K-Akt pathway and MAPK signalling in UVB-irradiated B16F10 melanoma cells.

<p>A, western blot analysis of B-raf, p-MEK, MEK1/2, p-ERK, ERK1/2, phospho-p38, p-38, phospho-JNK and JNK in cells treated with Qu and/or UVB. β-actin was used as loading control. B, C, D, E and F represent the densitometric analysis of B-raf, p-MEK/MEK, p-ERK/ERK, phospho-p38/p-38, phospho-JNK/JNK respectively. G, immunoblot analysis of PI3K-α and p-Akt in B16F10 cells at 24 h post-UVB and/ or Qu treatment. Signals were quantified for PI3K (H) and p-Akt (I) using Image Lab Software (Bio Rad). *, P<0.05; **, P<0.01 for control versus treatments; #, P<0.05, ##, P<0.01 for control versus UVB-alone treatment versus UVB + Qu treatments.</p

Effect of ascorbic acid (1 mM) on intracellular Ca²⁺ changes in response to treatment of UVB-irradiated B16F10 cells with quercetin.

<p>A, B16F10 cells were pre-treated with Qu for 24 h and supplemented with ascorbic acid 1 h before UVB irradiation. Following UVB irradiation, cells were loaded with Fluo-3 AM and analyzed on Olympus Flowview FV1000. B, represents the densitometric measure of the effect of ascorbic acid (AA) on intracellular Ca²⁺ levels in response to treatment of UVB-irradiated B16F10 cells with Qu.</p

Quercetin elevates UVB-induced reactive oxygen formation and intracellular calcium ion level.

<p>A, analysis of reactive oxygen species (ROS) formation immediately after UVB irradiation in B16F10 cells pre-treated with Qu for 24 hours. The generation of ROS was measured using BD FACS Calibur Aria. B, represents the fold increase in DCF fluorescence (a measure of ROS formation) relative to control. *, P<0.05; **, P<0.01 for control versus treatments; #, P<0.05, ##, P<0.01 for control versus UVB-alone treatment versus UVB + Qu treatments. C, effect of ascorbic acid (1 mM) on cell viability in response to treatment of UVB–irradiated B16F10 cells with Qu; UVB control cells were taken as 100% viable. D, analysis of intracellular free Ca²⁺ immediately after UVB exposure in B16F10 cells pre-treated with Qu for 24 hours. E, represents the effect of UVB (5 mJ/cm²) on calcium elevation in BAPTA-AM preloaded B16F10 cells. F, represents the fold increase in intracellular free calcium relative to control. *, P<0.05; **, P<0.01 for control versus treatments; #, P<0.05, ##, P<0.01 for control versus UVB-alone treatment versus UVB + Qu treatments.</p

Quercetin promotes UVB-induced cell death.

<p>A, structure of quercetin (<b>Qu</b>). B, analysis of cell viability using the MTT assay in B16F10 cells at 24 h post-UVB irradiation. <i>Columns</i>, mean of three experiments; <i>bars</i>, SD. *, P<0.05; **, P<0.01 for control versus treated. C, analysis of cell viability in Hs68 human fibroblast cells at 24 h post-UVB irradiation. D, analysis of cell viability using the MTT assay in B16F10 cells treated with Qu. *, P<0.05; **, P<0.01 for control versus Qu treated. E, analysis of cell viability using the MTT assay in human HaCaT keratinocytes at 24 h post-Qu treatment. F, analysis of cell viability using the MTT assay in Hs68 human fibroblast cells at 24 h post-Qu treatment. G, analysis of cell viability using the MTT assay in B16F10 cells treated with Qu and/or UVB (5 mJ/cm²) for 24 hours. *, P<0.05; **, P<0.01 for control versus treated; #, P<0.05; ##, P<0.01 for UVB-alone treatment versus UVB + Qu treatments. H, analysis of cell viability using the MTT assay in A375 human melanoma cells at 24 h post-Qu treatment. I, analysis of cell viability using the MTT assay in A375 cells treated with Qu and/or UVB (5 mJ/cm²) for 24 hours. J, analysis of cell viability in Hs68 cells treated with Qu and/or UVB for 24 hours.</p