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Epigenetics has emerged as a potent field of study for understanding the factors influencing the effectiveness of human disease treatments and for identifying alternations induced by pathogens in host plants. However, there has been a paucity of research on the epigenetic control of the proliferation and pathogenicity of fungal plant pathogens. Fungal plant pathogens such as Magnaporthe oryzae , a significant threat to global rice production, provide an important model for exploring how epigenetic mechanisms govern fungal proliferation and virulence. In M. oryzae , epigenetic alterations, such as DNA methylation, histone modification, and non-coding RNAs, regulate gene expression patterns that influence the pathogen's ability to infect its host. These modifications can enhance fungal adaptability, allowing the pathogen to survive in diverse environments and evade host immune responses. Our primary objective is to provide a comprehensive review of the existing epigenetic research on M. oryzae and shed light on how these changes influence the pathogen's lifecycle, its ability to invade host tissues, and the overall severity of the disease. We begin by examining the epigenetic alterations occurring in M. oryzae and their contributions to the virulence and proliferation of the fungus. To advance our understanding of epigenetic mechanisms in M. oryzae and similar plant diseases, we emphasize the need to address unanswered questions and explore future research directions. This information is crucial for developing new antifungal treatments that target epigenetic pathways, which could lead to improved disease management. [ABSTRACT FROM AUTHOR]

Onion (Allium cepa L.) and garlic (Allium sativum L.) are significant bulbous vegetables well known for their kitchen and medicinal values across the globe including Pakistan. Allium crops are highly vulnerable to many notorious RNA viruses fit in different genera viz., Allexivirus, Potyvirus, Tospovirus and Carlavirus. These viruses are known to inflict colossal and havoc yield losses up to 50% in allium crops wherever they grown. The occurrence of allium viruses in Pakistan has already been reported, but in single crop and only a few studies describe their incidence and distribution. The study examined the current scenario and distribution of major RNA viruses infecting Allium crops in Punjab, Pakistan. Field surveys were carried out in 14 districts of Punjab province as well as capital territory Islamabad of Pakistan, for two consecutive growing seasons (2019 and 2020). The 3000 leaf samples collected, 1675 samples (Onion 791, Garlic 884) were found positive for RNA viruses by using specie specific DAS-ELISA and further confirmed through reverse transcriptase polymerase chain reaction (RT-PCR). The results revealed the presence of RNA viruses with overall disease incidence of 52.7% and the highest disease incidence was observed as Onion yellow dwarf virus (OYDV) 58% following Irish yellow spot virus (IYSV) 41.8% respectively in onion crop during 2019-2020. Similarly, in garlic crop overall disease incidence of RNA viruses was noticed as 58.9% and the highest disease incidence of OYDV was observed as 45%, following IYSV 32.4% and GarV-C 22.5% respectively during 2019-2020. Comparatively, disease incidence and geographical distribution of viruses was variable in surveyed regions of both crops during surveyed period, whereas OYDV and IYSV were most prevalent viruses infecting onion and garlic crops and GarV-C found only in garlic. The current study also concluded the importance to disseminate the information/awareness among the farmers regarding viral diseases and their management by using different strategies including use of resistant genotypes. [ABSTRACT FROM AUTHOR]

Potato leafroll virus (PLRV) is a widespread and one of the most damaging viral pathogens causing significant quantitative and qualitative losses in potato worldwide. The current knowledge of the geographical distribution, standing genetic diversity and the evolutionary patterns existing among global PLRV populations is limited. Here, we employed several bioinformatics tools and comprehensively analyzed the diversity, genomic variability, and the dynamics of key evolutionary factors governing the global spread of this viral pathogen. To date, a total of 84 full-genomic sequences of PLRV isolates have been reported from 22 countries with most genomes documented from Kenya. Among all PLRV-encoded major proteins, RTD and P0 displayed the highest level of nucleotide variability. The highest percentage of mutations were associated with RTD (38.81%) and P1 (31.66%) in the coding sequences. We detected a total of 10 significantly supported recombination events while the most frequently detected ones were associated with PLRV genome sequences reported from Kenya. Notably, the distribution patterns of recombination breakpoints across different genomic regions of PLRV isolates remained variable. Further analysis revealed that with exception of a few positively selected codons, a major part of the PLRV genome is evolving under strong purifying selection. Protein disorder prediction analysis revealed that CP-RTD had the highest percentage (48%) of disordered amino acids and the majority (27%) of disordered residues were positioned at the C-terminus. These findings will extend our current knowledge of the PLRV geographical prevalence, genetic diversity, and evolutionary factors that are presumably shaping the global spread and successful adaptation of PLRV as a destructive potato pathogen to geographically isolated regions of the world. [ABSTRACT FROM AUTHOR]

Simple Summary: Growth trait is one of the most important economic traits for meat animals. Somatostatin DNA vaccine has been proven to enhance the growth rate of animals. However, the growth-promoting effect is not ideal. This study aimed to evaluate the immune effects of a novel eukaryotic dual expression vaccine known as pIRES-S/CST14-S/2SS in mice. Firstly, we demonstrated pIRES-S/CST14-S/2SS could functionally express in GH3 pituitary cells by regulating the growth hormone (GH) and prolactin (PRL) productions. Secondly, we observed that all concentrations of pIRES-S/CST14-S/2SS vaccine could evoke anti-somatostatin (SS) antibodies, leading to a higher level of GH concentration. Notably, pIRES-S/CST14-S/2SS (10 μg/100 μL) immunized mice obtained maximum body weight gain in a booster vaccination period. Our results are helpful for better understanding of the relationship between SS and CST, and pIRES-S/CST14-S/2SS vaccine may be used as a promising alternative for developing growth-promoting vaccine. Animal growth traits are directly linked with the economics of livestock species. A somatostatin DNA vaccine has been developed to improve the growth of animals. However, the growth-promoting effect is still unsatisfying. The current study aimed to evaluate the effect of a novel eukaryotic dual expression vaccine known as pIRES-S/CST14-S/2SS, which encodes the genes obtained by fusing somatostatin (SS) and cortistatin (CST) into hepatitis B surface antigen (HBsAg). After transfection into GH3 cells with pIRES-S/CST14-S/2SS, green fluorescence signals were observed by fluorescence microscopy, suggesting the effective expression of CST and SS in GH3 cells using the IRES elements. Subsequently, both GH and PRL levels were found to be significantly lower in pIRES-S/CST14-S/2SS-treated cells as compared to the control group (p < 0.05). Furthermore, the antibody level, hormone secretion, and weight gain in the mice injected with novel recombinant plasmids were also evaluated. The anti-SS antibodies were detectable in all vaccine treated groups, resulting in significantly higher levels of GH secretion (p < 0.05). It is worth mentioning that pIRES-S/CST14-S/2SS (10 μg/100 μL) vaccinated mice exhibited a higher body weight gain in the second immunization period. This study increases the understanding of the relationship between somatostatin and cortistatin, and may help to develop an effective growth-promoting DNA vaccine in animals. [ABSTRACT FROM AUTHOR]

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The small ubiquitin-related modifiers (SUMO-1/2/3) have been in the limelight as crucial dictators of a spectrum of biological events including normal growth and development. SUMO-1 protein is important for post-translational modification which is required for a variety of biological processes including organ development and organelle biogenesis. However, its functional relevance in mammalian ovary still remains to be explored. Herein, we found that SUMO-1 consistently localizes in granulosa cells (GCs), oocytes and corpus luteum during mice folliculogenesis. Moreover, the SUMO-1 conjugates particularly those which molecular weight correspond to 34-, 43-, and 72 KDa were highly presented in ovaries and were dynamically influenced by PMSG and/or hCG administration, further buttressing that SUMO-1 was expressed in mouse ovary. Additionally, mutation at G96/97A (binding site) changed the localization of SUMO-1 from nucleus to both cytoplasm and nucleus. With some diffusion from nucleus into cytoplasm, and the apoptosis in GCs was significantly triggered by overexpression of both SUMO-1 and its mutant SUMO-1G96/97A with more significantly induced by the latter form. Over-expression of wild-type SUMO-1 and mutant SUMO-1G96/97A significantly induced (P<0.001) apoptosis in GCs, but the mutant SUMO-1 induced higher apoptotic rate (P<0.001) than over expressed SUMO-1. Overall, our study determines the expression of SUMO-1 in mouse ovary and pinpoints SUMO-1 as a determinant of GCs biology which is essential for mammalian ovary function. [ABSTRACT FROM AUTHOR]

This study was aimed to address melatonin receptor expression, mRNA level of hypothalamus and hypophysis hormone receptors (GnRHR, FSHR, and LHR), steroidogenesis, cell cycle, apoptosis, and their regulatory factors after addition of melatonin for 24 hr in cultured buffalo granulosa cells (GCs). The results revealed that direct addition of different concentrations of melatonin (100 pM, 1 nM, and 100 nM) resulted in significant upregulation (p < 0.05) of mRNA level of melatonin receptor 1a (MT1) without affecting melatonin receptor 1b (MT2). Melatonin treatment significantly downregulated (p < 0.05) mRNA level of FSH and GnRH receptors, whereas 100 nM dose of melatonin significantly increased mRNA level of LH receptor. Treatment with 100 nM of melatonin significantly decreased the basal progesterone production with significant decrease (p < 0.05) in mRNA levels of StAR and p450ssc, and lower mRNA level of genes (Insig1, Lipe, and Scrab1) that affect cholesterol availability. Melatonin supplementation suppressed apoptosis (100 nM, p < 0.05) and enhanced G2/M phase (1 nM, 100 nM, p < 0.05) of cell cycle progression which was further corroborated by decrease in protein expression of caspase‐3, p21, and p27 and increase in bcl2. Our results demonstrate that melatonin regulates gonadotrophin receptors and ovarian steroidogenesis through MT1. Furthermore, the notion of its incorporation in apoptosis and proliferation of buffalo GCs extends its role in buffalo ovaries. [ABSTRACT FROM AUTHOR]

The present report accounts successful production of blastocysts and birth of calves following OPU-IVEP technique in Sahiwal cows of Pakistani origin. A total of 9 transvaginal OPU sessions were performed in twelve Sahiwal cows and 520 immature COCs were recovered using commercially prepared media. These COCs were placed in maturation medium for 24 h in an incubator at 38.5°C with 5% CO2. The matured oocytes were co-incubated with sperm cells in IVF medium for 22 h. After fertilization, the presumptive zygotes were transferred to culture medium for 7 days at 5% CO2. The results showed that the cleavage rate of IVF was 68.6% out of which 26.3% were developed up to blastocyst. A total of 2 blastocyst-stage fresh embryos were transferred in a single synchronized recipient cow. The pregnancy was confirmed one month after transfer and resulted in successful calving demonstrating the possibility of performing embryo transfer in elite native herds. [ABSTRACT FROM AUTHOR]

The objectives of this study are to find out the effect of melatonin treatments (43μM & 430μM) on mRNA and protein expression of casein, and melatonin receptors in buffalo mammary epithelial cells (BuMEC) using reverse transcriptional polymerase chain reaction (qRT-PCR) and Western blotting. Furthermore, by using specific inhibitors against mTOR (Rapamycin) and Jak2-Stat5 pathway (AG490), effect of melatonin treatment on translational level of casein was measured. The results showed that melatonin treatment (430 μM) significantly up-regulated casein level both at transcriptional (P<0.001) and translational (P<0.05) levels. Melatonin treatment significantly up-regulated mRNA expression of (P<0.05) melatonin receptor 1a (MT1) but melatonin receptor 1b (MT2) did not significantly varied. In the presence of mTOR inhibitor, rapamycin, melatonin failed to up regulate (P<0.05) protein expression of casein but no significant difference was found in the level of casein in the presence of Jak2-Stat5 inhibitor (AG490). These results indicate the significance of melatonin in altering milk casein expression in mammary epithelial cells in buffalo. Keeping in view the seasonality of buffalo in tropical and subtropical region, this study is important in the establishment of melatonin as the key modulator in changing concentration of casein in milk. [ABSTRACT FROM AUTHOR]

Objective: Identification of the candidate genes that play key roles in phenotypic variations can provide new information about evolution and positive selection. Interleukin (IL)-32 is involved in many biological processes, however, its role for the immune response against various diseases in mammals is poorly understood. Therefore, the current investigation was performed for the better understanding of the molecular evolution and the positive selection of single nucleotide polymorphisms in IL-32 gene. Methods: By using fixation index (FST) based method, IL-32 (9375) gene was found to be outlier and under significant positive selection with the provisional combined allocation of mean heterozygosity and FST. Using nucleotide sequences of 11 mammalian species from National Center for Biotechnology Information database, the evolutionary selection of IL-32 gene was determined using Maximum likelihood model method, through four models (M1a, M2a, M7, and M8) in Codeml program of phylogenetic analysis by maximum liklihood. Results: IL-32 is detected under positive selection using the FST simulations method. The phylogenetic tree revealed that goat IL-32 was in close resemblance with sheep IL-32. The coding nucleotide sequences were compared among 11 species and it was found that the goat IL-32 gene shared identity with sheep (96.54%), bison (91.97%), camel (58.39%), cat (56.59%), buffalo (56.50%), human (56.13%), dog (50.97%), horse (54.04%), and rabbit (53.41%) respectively. Conclusion: This study provides evidence for IL-32 gene as under significant positive selection in goat. [ABSTRACT FROM AUTHOR]

Hepatic fibrosis is a common feature of chronic liver injury, and the involvement of angiotensin II in such process has been studied earlier. We hypothesized that anti-angiotensin II agents may be effective in preventing hepatic fibrosis. In this study, Long Evans female rats were used and divided into four groups such as Group-I, Control; Group-II, Control + ramipril; Group-III, CCl4; and Group-IV, CCl4 + ramipril. Group II and IV are treated with ramipril for 14 d. At the end of treatment, the livers were removed, and the level of hepatic marker enzymes (aspartate aminotransferase, Alanine aminotransferase, and alkaline phosphatase), nitric oxide, advanced protein oxidation product , catalase activity, and lipid peroxidation were determined. The degree of fibrosis was evaluated through histopathological staining with Sirius red and trichrome milligan staining. Carbon-tetrachloride (CCl4) administration in rats developed hepatic dysfunction and raised the hepatic marker enzymes activities significantly. CCl4administration in rats also produced oxidative stress, inflammation, and fibrosis in liver. Furthermore, angiotensinogen-inhibitor ramipril normalized the hepatic enzymes activities and improved the antioxidant enzyme catalase activity. Moreover, ramipril treatment ameliorated lipid peroxidation and hepatic inflammation in CCl4-treated rats. Ramipril treatment also significantly reduced hepatic fibrosis in CCl4-administered rats. In conclusion, our investigation suggests that the antifibrotic effect of ramipril may be attributed to inhibition of angiotensin-II mediated oxidative stress and inflammation in liver CCl4-administered rats. [ABSTRACT FROM PUBLISHER]

Citrus maxima peel is rich in natural phenolic compounds and has a long use in the traditional medicine. HPLC-DAD analysis on Citrus maxima peel powder exhibited the presence of various phenolic compounds such as caffeic acid and (−)-epicatechin. To determine the plausible hepatoprotective activity of Citrus maxima peel powder, we used carbon tetrachloride (CCl4) treated rat model. Liver damage in rats was confirmed by measuring the AST, ALT, and ALP enzyme activities. In addition, lipid peroxidation products (MDA), nitric oxide, advanced protein oxidation products level (APOP), and catalase activities were also analyzed along with the histological profiling for the inflammatory cell infiltration, collagen, and iron deposition in liver. Dietary supplementation of Citrus maxima peel powder exhibited significant reduction of serum AST, ALT, and ALP activities in carbon tetrachloride treated rats. Moreover, Citrus maxima peel powder also showed a significant reduction of the oxidative stress markers (MDA, NO, and APOP level) and restored the catalase activity in CCl4 treated rats. Histological examination of the liver section revealed reduced inflammatory cells infiltration, collagen, and iron deposition in CCl4 treated rats. The results from this study demonstrated that Citrus maxima peel powder produced significant hepatoprotective action in CCl4 administered rats. [ABSTRACT FROM AUTHOR]

Background: Ucche (Momordica charantia L. var. muricata (Willd.) Chakravarty) has been reported to possess many benefits and medicinal properties. However, the protective effect of ucche against carbon tetrachloride (CCl4) induced hepatotoxicity have not been clarified fully yet. The aim of the present study was to investigate the effects of ucche on oxidative stress and inflammation in liver of CCl4 treated rats. Methods: Female Long Evans rats were administered with CCl4 orally (1 ml/kg) twice a week for 2 weeks and were supplemented with freshly prepared crashed ucche (10% wt/wt of diet) with powdered chaw food. Both plasma and liver tissues were analyzed for AST, ALT and ALP activities. Oxidative stress parameters were measure by determining malondialdehyde (MDA), nitric oxide (NO), advanced protein oxidation product (APOP), and reduced glutathione (GSH) concentrations and catalase activities in plasma and liver tissues. Moreover, inflammation and tissue fibrosis were confirmed by histological staining of liver tissue sections. Results: Our data suggest that ucche significantly prevented CCl4-induced hepatotoxicity, indicated by both diagnostic indicators of liver damage (serum transferases activities) and histopathological analysis. Moreover, CCl4 administration induced profound elevation of reactive oxygen species (ROS) production and oxidative stress, as evidenced by increasing lipid peroxidation level and depletion of antioxidant enzymes in liver. Fresh ucche supplementation prevented the oxidative stresses and improved antioxidant enzyme function. Furthermore, fresh ucche supplementation reduced hepatic inflammatory cell infiltration, iron deposition and fibrosis in liver of CCl4 treated rats. Conclusion: In conclusion, these results suggested that the inhibition of CCl4-induced inflammation by ucche is due at least in part to its anti-oxidant activity and its ability to modulate the inflammation and fibrosis in liver. [ABSTRACT FROM AUTHOR]

Background: Ucche (Momordica charantia L. var. muricata (Willd.) Chakravarty) has been reported to possess many benefits and medicinal properties. However, the protective effect of ucche against carbon tetrachloride (CCl4) induced hepatotoxicity have not been clarified fully yet. The aim of the present study was to investigate the effects of ucche on oxidative stress and inflammation in liver of CCl4 treated rats. Methods: Female Long Evans rats were administered with CCl4 orally (1 ml/kg) twice a week for 2 weeks and were supplemented with freshly prepared crashed ucche (10% wt/wt of diet) with powdered chaw food. Both plasma and liver tissues were analyzed for AST, ALT and ALP activities. Oxidative stress parameters were measure by determining malondialdehyde (MDA), nitric oxide (NO), advanced protein oxidation product (APOP), and reduced glutathione (GSH) concentrations and catalase activities in plasma and liver tissues. Moreover, inflammation and tissue fibrosis were confirmed by histological staining of liver tissue sections. Results: Our data suggest that ucche significantly prevented CCl4-induced hepatotoxicity, indicated by both diagnostic indicators of liver damage (serum transferases activities) and histopathological analysis. Moreover, CCl4 administration induced profound elevation of reactive oxygen species (ROS) production and oxidative stress, as evidenced by increasing lipid peroxidation level and depletion of antioxidant enzymes in liver. Fresh ucche supplementation prevented the oxidative stresses and improved antioxidant enzyme function. Furthermore, fresh ucche supplementation reduced hepatic inflammatory cell infiltration, iron deposition and fibrosis in liver of CCl4 treated rats. Conclusion: In conclusion, these results suggested that the inhibition of CCl4-induced inflammation by ucche is due at least in part to its anti-oxidant activity and its ability to modulate the inflammation and fibrosis in liver. [ABSTRACT FROM AUTHOR]

Background Vanda tessellata (Orchidaceae) has been used in different sorts of ailments such as inflammations, rheumatism, dysentery, bronchitis, dyspepsia and fever in folk medicine. In this study we evaluated the antinociceptive and cytotoxic effect of methanol and aqueous extracts of V. tessellata leaf. Methods Oral administration of V. tessellata aqueous (VTA) and methanol (VTM) leaf extracts at 200 and 400 mg/kg body weight (bw) doses were assessed for antinociceptive activity in acetic acid-induced writhing test, hot plate test, and tail immersion test in mice. In this study we also screened for cytotoxicity of the extracts by the brine shrimp (Artemia salina) lethality assay. Results The extracts, at both doses, exhibited a significant (p < 0.05 - < 0.01) dose-dependent antinociceptive activity in hot plate and tail immersion test. The reaction time was increased to the thermal stimuli at 200 and 400 mg/kg doses up to 90 min. In acetic acid-induced writhing test, oral administration of VTA and VTM (200 and 400 mg/kg) also decreased the writhing significantly while compared to control. The dose 400 mg/kg showed maximum percentage of pain inhibition 42.37% and 45.08% for VTA and VTM respectively. Diclofenac sodium (10 mg/kg) and nalbuphine (10 mg/kg) were used as reference antinociceptive drugs. Very low cytotoxicity was observed on brine shrimps lethality assay. Conclusions The leaf extract has potential antinociceptive activity with minimum cytotoxicity. The present study supports the use of V. tessellata in different inflammatory disorders. [ABSTRACT FROM AUTHOR]

The objective of this study was to examine the efficacy and safety of a novel inhibin vaccine containing inhibin α (1-32) fragments in mice. A recombinant plasmid pVAX-asd-IS was constructed by inserting recombinant inhibin α (1-32) and the hepatitis B surface antigen S into the plasmid in which the asd gene, rather than the kanamycin gene, was a selection marker. Ninety Kuming mice were divided into six groups consisting of 15 mice each. First group was (C1) injected with 200 µl of PBS, second (C2) received 1 × 1010 CFU of crp −/ asd − C500/pVAX-asd and served as vector control, third did not receive any treatment (C3), while fourth, fifth, and sixth group received 1 × 1010, 1 × 109, 1 × 108 CFU of the recombinant inhibin vaccine crp −/ asd − C500/pVAX-asd-IS (group T1, T2, T3), respectively. Western blotting demonstrated that recombinant expressed inhibin protein possessed immune function and that this plasmid could replicate for up to 40 generations stably. Vaccination with this strain at a dose of 1 × 1010 CFU/200 µl per mouse induced high anti-inhibin antibody levels, significantly increased large-follicle production in T1 group ( p < 0.05) and average litter size ( p > 0.05) compared with control groups. Integration studies showed no evidence of inhibin fusion gene integrated into mice's genome 2-month after immunization. These results suggest that the vaccine described in the present study may provide a safe method to improve reproductive traits in animals. A trend towards increased litter size and significant increase in large follicle population depict that this vaccine may have direct application in large animal industry. [ABSTRACT FROM AUTHOR]

Low cloning efficiency is considered to be caused by the incomplete or aberrant epigenetic reprogramming of differentiated donor cells in somatic cell nuclear transfer (SCNT) embryos. Oxamflatin, a novel class of histone deacetylase inhibitor (HDACi), has been found to improve the in vitro and full-term developmental potential of SCNT embryos. In the present study, we studied the effects of oxamflatin treatment on in vitro porcine SCNT embryos. Our results indicated that the rate of in vitro blastocyst formation of SCNT embryos treated with 1 μM oxamflatin for 15 h postactivation was significantly higher than all other treatments. Treatment of oxamflatin decreased the relative histone deacetylase (HDAC) activity in cloned embryos and resulted in hyperacetylation levels of histone H3 at lysine 9 (AcH3K9) and histone H4 at lysine 5 (AcH4K5) at pronuclear, two-cell, and four-cell stages partly through downregulating HDAC1. The suppression of HDAC6 through oxamflatin increased the nonhistone acetylation level of α-tubulin during the mitotic cell cycle of early SCNT embryos. In addition, we demonstrated that oxamflatin downregulated DNA methyltransferase 1 ( DNMT1) expression and global DNA methylation level (5-methylcytosine) in two-cell-stage porcine SCNT embryos. The pluripotency-related gene POU5F1 was found to be upregulated in the oxamflatin-treated group with a decreased DNA methylation tendency in its promoter regions. Treatment of oxamflatin did not change the locus-specific DNA methylation levels of Sus scrofa heterochromatic satellite DNA sequences at the blastocyst stage. Meanwhile, our findings suggest that treatment with HDACi may contribute to maintaining the stable status of cytoskeleton-associated elements, such as acetylated α-tubulin, which may be the crucial determinants of donor nuclear reprogramming in early SCNT embryos. In summary, oxamflatin treatment improves the developmental potential of porcine SCNT embryos in vitro. [ABSTRACT FROM AUTHOR]

The present study was designed to compare the effect of different doses of equine chorionic gonadotropin (eCG) on follicular population and plasma estradiol concentration in Chinese Holstein cows synchronized with prostaglandin F2a (PGF2a). One hundred cows were divided into five treatment groups. Each group (n=20) was administered with an injection of eCG at dose rate of 2, 2.5, 3, 3.5, or 4 IU/kg b.w., respectively, followed by administration of PGF2a 48 h later. Ovarian scanning was performed by transrectal ultrasonography around estrus, while plasma estradiol concentration at administration of eCG, PGF2a, estrus and 12 h after estrus was measured by Radioimmunoassay (RIA). The results showed that medium follicles (5-10 mm) varied (P<0.05) among all treatment groups with an increasing trend. However, there was no significant difference (P>0.05) in the number of large follicles (>10 mm). Plasma estradiol concentration differed non-significantly (P>0.05) in all groups before estrus. At estrus, the concentration was at peak in high dose treatment group (4 IU/kg, P<0.05) than those of low dose treatment groups (2 and 2.5 IU/kg). The study concluded that eCG increases the recruitment and development of medium follicles in dose dependent manners however it has no subsequent effect on large follicles population. © 2013 Friends Science Publishers [ABSTRACT FROM AUTHOR]

Histone acetylation is one of the most important posttranslational modifications that contribute to transcriptional initiation and chromatin remodeling. In our previous study, we enhanced sperm chromatin remodeling within the bovine sperm injection-derived androgenentic (SpI-AG) embryos by sperm pretreatment, and thereby improved their early developmental competence. In this study, we found that blastocyst development of SpI-AG embryos could be elevated by the histone deacetylase inhibitor (HDACi). First, we optimized the efficacy of two histone deacetylase inhibitors [trichostatin A (TSA) and Scriptaid (SCR)] in a dose (0, 5, 10, 20, 50, and 100 nM for TSA; 0, 50, 100, 200, 300, and 500 nM for SCR, respectively) and time-dependent (0, 10, 15, 20, and 25 h) manner on the developmental capacity of these embryos. Furthermore, we quantitatively assessed the alterations in histone H3 and H4 overall acetylation levels and blastocyst quality of SpI-AG embryos by immunofluorescence staining. We found a significantly improved morula and blastocyst development rate of SpI-AG embryos at a mild dose of TSA (20 nM) or SCR (200 nM) for 15 h after embryo activation. Furthermore, both HDACi noticeably increased the levels of acetylated histone H3 and H4 in SpI-AG blastocyst embryos, whereas, SCR treatment improved the quality of blastocysts when compared with control group. In conclusion, HDACi is beneficial for early development of bovine SpI-AG embryos and can be used to improve the efficiency of its in vitro production. [ABSTRACT FROM AUTHOR]

Inhibin, a member of the transforming growth factor-β [TGF-β] superfamily, is a suppressor of follicle-stimulating hormone [FSH] release through pituitary–gonadal negative feedback loop to regulate follicular development. In this study, Inhibin α-subunit [Inha] gene was knocked down successfully in mice primary anterior pituitary cells at both transcriptional and translational levels by RNAi-Ready pSIREN-RetroQ-ZsGreen Vector mediated recombinant pshRNA vectors. The results indicated that inhibin silencing significantly promoted apoptosis by up-regulating Caspase-3, Bax and Bcl-2 genes without affecting p53 both at transcriptional and translational levels. Furthermore, it markedly impaired the progression of G1 phase of cell cycle and decreased the amount of cells in S phase [as detected by flow cytometry]. Inhibin silencing resulted in significant up-regulation of mRNA and protein expressions of Gondotropin releasing hormone receptors [GnRHR] and down-regulated mRNA levels of β-glycans with parellel change in the amount of its protein expression. Silencing of inhibin-a significantly increased [P<0.05] activin-β concentration without affecting FSH and LH levels in anterior pituitary cells. These findings revealed that up regulation of GnRH receptors by silencing inhibin a-subunit gene might increase the concentration of activin-β in the culture medium. Inhibin a silencing resulted in increased mRNA and protein expressions of inhibinβ which may demonstrate that both inhibin subunits co-participate in the regulation of reproductive events in anterior pituitary cells. This study concludes that inhibin is a broad regulatory marker in anterior pituitary cells by regulating apoptosis, cellular progression and simultaneously by vital fluctuations in the hormonal signaling. [ABSTRACT FROM AUTHOR]

Diabetic glomerulosclerosis is characterized by accumulation of extracellular matrix proteins, mesangial expansion, and tubulointerstitial fibrosis. Hyperglycemia accelerates development of the disease, a direct result of increased intracellular glucose availability. The facilitative glucose transporter GLUT1 mediates mesangial cell glucose flux which leads to activation of signaling cascades favoring glomerulosclerosis, including pathways mediated by angiotensin II (Ang II), transforming growth factor β (TGF-β), connective tissue growth factor (CTGF), and vascular endothelial growth factor (VEGF). Ang II has both hemodynamic and metabolic effects directly inducing GLUT1 and/or matrix protein synthesis through diacyl glycerol (DAG) or protein kinase C (PKC) induction, mesangial cell stretch, and/or through transactivation of the epidermal growth factor receptor, the platelet-derived growth factor receptor, and the insulin-like growth factor-1 receptor, all of which may stimulate GLUT1 synthesis via an ERK-mediated pathway. Conversely, inhibition of Ang II effects suppresses GLUT1 and cellular glucose uptake. GLUT1-mediated glucose flux leads to metabolism of glucose via glycolysis, with induction of DAG, PKC, TGF-β1, CTGF and VEGF. VEGF in turn triggers both GLUT1 and matrix synthesis. New roles for GLUT1-mTOR and GLUT1-mechano-growth factor interactions in diabetic glomerulosclerosis have also recently been suggested. Recent mouse models confirmed roles for GLUT1 in vivo in stimulating glomerular growth factor expression, growth factor receptors and development of glomerulosclerosis. GLUT1 may therefore act in concert with cytokines and growth factors to induce diabetic glomerulosclerosis. Further clarification of the pathways involved may prove useful for the therapy of diabetic nephropathy. New directions for investigation are discussed. Copyright © 2013 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Fasciolosis is an economically important disease for livestock, as well as being zoonotic. Recent figures on the prevalence of this disease have caused alarm concerning its potential for an increased prevalence in the future. The prevalence of fascioliosis has been documented from different regions of the world, helping us identify areas where future research needs to be focused. This manuscript is a review of the current status of the disease, the pathogenic species involved, diagnostic techniques (with new modifications and comparative specificity, sensitivity, and rapidity of these tests), chemotherapy, and vaccination. This also encompasses inaccurate reports on vaccination and drug development as well as the latest technologies to find promising candidates for drugs and vaccines. Drugs with lower efficacy have been used on some farms which lead to exacerbation of the clinical disease, presumably due to the development of drug resistance. Future studies should be focused on (1) the use of the most reliable diagnostic tests for periodic monitoring of the disease, (2) insights of the ecobiology and transmission dynamics of the snail intermediate host and the best possible methods of their control, (3) in vitro and in vivo testing of chemotherapeutic compounds using sensitive methods, and (4) the identification of novel drug and vaccine candidates using modern molecular markers. This approach may help increase the reliability of chemotherapeutic agents and control nuisance, ultimately reducing the economic losses attributable to the livestock industry around the world. [ABSTRACT FROM AUTHOR]

In recent decades, Pakistan has suffered a decline in cotton production due to several factors, including insect pests, cotton leaf curl disease (CLCuD), and multiple abiotic stresses. CLCuD is a highly damaging plant disease that seriously limits cotton production in Pakistan. Recently, genome editing through CRISPR/Cas9 has revolutionized plant biology, especially to develop immunity in plants against viral diseases. Here we demonstrate multiplex CRISPR/Cas-mediated genome editing against CLCuD using transient transformation in N. benthamiana plants and cotton seedlings. The genomic sequences of cotton leaf curl viruses (CLCuVs) were obtained from NCBI and the guide RNA (gRNA) were designed to target three regions in the viral genome using CRISPR MultiTargeter. The gRNAs were cloned in pHSE401/pKSE401 containing Cas9 and confirmed through colony PCR, restriction analysis, and sequencing. Confirmed constructs were moved into Agrobacterium and subsequently used for transformation. Agroinfilteration in N. benthamiana revealed delayed symptoms (3–5 days) with improved resistance against CLCuD. In addition, viral titer was also low (20–40%) in infected plants co-infiltrated with Cas9-gRNA, compared to control plants (infected with virus only). Similar results were obtained in cotton seedlings. The results of transient expression in N. benthamiana and cotton seedlings demonstrate the potential of multiplex CRISPR/Cas to develop resistance against CLCuD. Five transgenic plants developed from three experiments showed resistance (60−70%) to CLCuV, out of which two were selected best during evaluation and screening. The technology will help breeding CLCuD-resistant cotton varieties for sustainable cotton production. [ABSTRACT FROM AUTHOR]

Due to higher fecundity rate over Beetal goats, it is assumed that Teddy goats have higher ovarian activity. Therefore, the current study tested the hypothesis that ovarian follicular population i.e. number of small (2-4 mm), medium (>4-6 mm) and large follicles (>6 mm), size of the ovulatory follicle, and ovulation rate (No. of CL/No of ovulated goats), using transrectal ultrasonography are different between Beetal and Teddy goats of Pakistani origin. Beetal (n=6) and Teddy (n=8) does were synchronized using double PGF2a injections 10 d apart and were scanned on Days -2, 0 (estrus) and +2. The onset of estrus was assessed by aproned bucks. Mean number of small follicles were higher (P<0.05) in Beetal goats, compared to Teddy goats, on days -2, estrus and +2. The ovulatory follicle diameter, and ovulation rate were found non-significant between Beetal and Teddy goats. It is concluded that Beetal goats have greater population of small follicles around estrus compared to Teddy goats. Future studies can be based on the follicle recruitment and atretic factors in Beetal goats to enhance the production. [ABSTRACT FROM AUTHOR]

Four blackbucks, 4-6 years of age were handled for dystocia with a history of straining and without successful delivery. Clinical examination revealed normal vital parameters, partially opened cervix and dry birth canal. Water bags were ruptured in all cases. In three out of four cases (75%), the fetus was extracted out using standard obstetrical operations; however, one fetus (25%) was taken out using fetotomy. All fetuses were dead after assisted delivery. Three out of four (75%) dams were found active and behaved normally after delivery, however, one dam (25%) died soon after obstetrical procedure. It was concluded that dystocia in blackbucks can be effectively handled on similar lines as in caprines and timely veterinary assistance is required in order to save the newborns. [ABSTRACT FROM AUTHOR]