Effect of different levels of single cell protein and probiotic microorganisms on performance, immunological responses, and intestinal histology in laying hens.

An experiment was conducted to evaluate the effects of single-cell protein (SCP), as a valuable by-product and substitute for soybean meal, and multistrain probiotics on the performance, blood biochemistry, intestinal histomorphology, ileal microbiota, and immune responses in laying hens. Leghorn laying hens were randomly assigned to nine experimental treatments with eight replicates of five birds each. Dietary treatments consisted of three levels of replacement of soybean meal with SCP (0.0%, 30%, and 60%) and three levels of probiotics (0.0, 50, 100 mg/kg) offered through 70 days of main recording. The result indicated that replacing 30% soybean meal with SCP in the diet significantly increased the eggshell weight, thickness, and strength; however, hens receiving 60% SCP replacement showed lower productive performance. During the second 35-d trial, hens receiving at least 50 mg/kg probiotics produced higher eggshell weight, thickness, and strength than the control hens. Remarkable lower levels of serum cholesterol content were observed in the SCP and probiotic-supplemented diets. The significant reduction in the ileal pH, Escherichia coli and Clostridium spp. enumerations and a remarkable increase in Lactobacillus count was observed after feeding incremental levels of probiotics. The highest villus height to crypt depth ratio, antibody titer against sheep red blood cells (in secondary response) and Newcastle disease vaccine, as well as egg production and egg mass, were found in the hens that received 30% SCP replacement in combination with 50 or 100 mg/kg probiotic (SCP × Probiotic). The present study concluded that feeding SCP, as a cheap protein alternative source, along with probiotics while reducing production costs can support productive performance via modulating the intestinal microbiomes and health indices and fortifying the immune system in laying hens.

Synergistic effects of punicic acid and alpha lipoic acid ameliorate inflammatory and metabolic genes expression in C2C12 myoblast cells under oxidative stress condition.

Inflammation is a reaction of the immune system to infection and injury; in fact, it positioned at the center of metabolic disorders, particularly obesity, type 2 diabetes, and cardiovascular diseases. Thus play a major role not only in their development, but also exerts as a crucial linking factor among those diseases. In this regard, one of the strategies for tackling this problem is application of antioxidants to treat such diseases. The present study was performed to evaluate the synergistic effects of punicic acid (PUA) and alpha-lipoic acid (ALA) as antioxidants and radical scavenging reagents on the expression of some inflammatory and metabolism-related genes under oxidative stress in the muscle cells. The experimental treatments consisted of a range of 20, 40, 80, 160, and 320 µM of PUA, and 5, 25, 50, 100, and 200 µM of ALA with a 200 µM concentration of H2 O2 as an oxidative stress inducer. Accordingly, fatty acid treatments were applied for 24 h, and H2 O2 was treated for 1 h. Our results indicated that the simultaneous treatment of PUA and ALA at optimal concentrations (80 and 50 µM, respectively) decreased the expression of inflammation genes and increased the expression of regulatory genes (Pparγ, Pgc-1α) related to metabolism (p < .05). Unexpectedly, H2 O2 treatment increased the Fndc5 expression (p < .05). Maximal upregulation of Pparγ, Pgc-1α were obtained when fatty acids combination (PUA and ALA) were used in the culture of H2 O2 treated cells (p < .05). Therefore, our findings suggest that the simultaneous use of PUA and ALA fatty acids could reduce oxidative stress, and the expression of inflammatory genes, thereby improving the cell metabolism.

Risk of mortality in COVID-19 patients: a meta- and network analysis.

Understanding the most relevant hematological/biochemical characteristics, pre-existing health conditions and complications in survivors and non-survivor will aid in predicting COVID-19 patient mortality, as well as intensive care unit (ICU) referral and death. A literature review was conducted for COVID-19 mortality in PubMed, Scopus, and various preprint servers (bioRxiv, medRxiv and SSRN), with 97 observational studies and preprints, consisting of survivor and non-survivor sub-populations. This meta/network analysis comprised 19,014 COVID-19 patients, consisting of 14,359 survivors and 4655 non-survivors. Meta and network analyses were performed using META-MAR V2.7.0 and PAST software. The study revealed that non-survivors of COVID-19 had elevated levels of gamma-glutamyl transferase and creatinine, as well as a higher number of neutrophils. Non-survivors had fewer lymphocytes and platelets, as well as lower hemoglobin and albumin concentrations. Age, hypertension, and cerebrovascular disease were shown to be the most influential risk factors among non-survivors. The most common complication among non-survivors was heart failure, followed by septic shock and respiratory failure. Platelet counts, creatinine, aspartate aminotransferase, albumin, and blood urea nitrogen levels were all linked to ICU admission. Hemoglobin levels preferred non-ICU patients. Lower levels of hemoglobin, lymphocytes, and albumin were associated with increased mortality in ICU patients. This meta-analysis showed that inexpensive and fast biochemical and hematological tests, as well as pre-existing conditions and complications, can be used to estimate the risk of mortality in COVID-19 patients.

An overview of the role of metallic and nonmetallic nanoparticles and their salts during sperm cryopreservation and in vitro embryo manipulation.

The cryopreservation of spermatozoa and the in vitro embryo production are valuable tools used in a variety of species, including humans, livestock, fish, and aquatic invertebrates. Sperm cryopreservation has been used to maintain or increase the genetic diversity of threatened species. Reactive oxygen species (ROS) are molecules derived from oxygen, being formed as byproducts of cellular metabolism. During cryopreservation of sperm and other in vitro manipulations of oocytes and embryos, ROS production is dramatically increased. In cells, low, medium, and high levels of ROS lead to different outcomes, apoptosis, auto-phagocytosis, and necrosis, respectively. ROS produced by cells can be neutralized by intracellular antioxidant systems, including enzymes as well as non-enzymatic antioxidants. Free radicals and oxidative stress can be major factors influencing in vitro manipulations. In this review, we discuss the role that metallic and nonmetallic nanoparticles and their salts play in the modulation of oxidative stress during in vitro embryo production and cryopreservation of sperm.

Effects of nomadic grazing system and indoor concentrate feeding systems on performance, behavior, blood parameters, and meat quality of finishing lambs.

The objective of the study was to evaluate the effects of three production systems on growth performance, behavior, blood parameters, carcass characteristics, and meat quality. A total of 30 lambs (n = 10 lambs/treatment) were randomly assigned to three production systems that included rotational grazing (NG) and two different levels of concentrate (CON), one with medium (roughage/concentrate ratio 50:50% based on DM, MC) and one with high concentrate (roughage/concentrate ratio 30:70% based on DM, HC) during the 90-day fattening period. At the start of the experiment, all lambs averaged 90 ± 4 days of age (mean ± SD) and were slaughtered at an average of 180 ± 3 days (mean ± SD). CON-fed lambs had higher average daily gain and loin thickness than NG-fed lambs. The NG lambs spent more time eating, drinking, and standing, but less time resting and rumination than the CON-fed lambs. In addition, plasma lipid, ß-hydroxybutyrate, and urea levels were higher, while phosphorus levels were lower in NG-fed lambs than in CON-fed lambs. CON-fed lambs had better carcass yield, but gastrointestinal tract and rumen weights were lower than NG lambs. CON-fed lambs had higher pH values 0 h post mortem than the NG lambs; however, there was no effect of treatment on pH 24 h post mortem. The post-mortem color of the LD muscle of NG lambs had a higher lightness and yellowness index and a lower redness index than that of the LD muscle of CON-fed lambs. The results of this study showed that lambs fed CON had better carcass yield than lambs fed NG, although feed intake, feed conversion ratio (FCR), growth performance, carcass yield, and meat quality of lambs fed MC and HC were similar.

Nutrient intake, digestibility, and serum metabolites in dairy cows fed diets differing in starch concentration with palmitic acid or stearic acid supplementation postpartum.

The present study determined the effect of supplementing palmitic acid (PA) and stearic acid (SA) on the nutrient intake, digestibility, and serum metabolites of dairy cows fed two different starch levels during the postpartum period. Forty-four multiparous Holstein cows were used in a completed randomized block based on their parity and previous milk yield. Dietary treatments were arranged in a 2 × 2 arrangement with two dietary starch levels (HS: 260 g/kg of diet dry matter (DM) vs LS: 210 g/kg of diet DM) and two fat supplements rich in PA or SA at 15 g/kg of diet DM. Increasing the starch concentration of the postpartum diet improved organic matter (OM), ether extract (EE), crude protein (CP), and starch intake. Moreover, HS diets resulted in higher apparent digestibility of OM and CP but lower starch digestibility than LS diets. Feeding HS diets increased fecal starch output compared with LS diets. There was starch levels and FA supplements interaction for serum albumin and total antioxidant capacity (TAC), with higher concentrations in HSSA and LSPA compared to HSPA and LSSA. Significant correlations between performance and blood metabolites were observed in weeks 3 and 4. In week 3, a negative correlation was observed between serum TAC with milk protein (r = - 0.51) and lactose percentage (r = - 0.49) in the HS diet. However, non-esterified FA was correlated with the fat to protein ratio in the LS diet (r = 0.54). Moreover, in week 4, serum TAC was negatively related to the body condition score of the cows fed LS diet (r = 0.50), while there was no relationship for cows fed HS diets. In conclusion, feeding HS diets to postpartum cows increased nutrient intake and the digestibility of OM and CP compared with LS diets. The addition of SA to the HS diet may be more beneficial than PA in improving the oxidative status of dairy cows in the postpartum period.

Polycaprolactone-co-polylactic acid nanofiber scaffold in combination with 5-azacytidine and transforming growth factor-ß to induce cardiomyocyte differentiation of adipose-derived mesenchymal stem cells.

Adipose-derived mesenchymal stem cells (Ad-MSCs) are promising candidates for cardiac repair/regeneration. The application of copolymer nanoscaffolds has received great attention in tissue engineering to support differentiation and functional tissue organization toward effective tissue regeneration. The objective of the current study was to develop functional and bioactive scaffolds by combining polycaprolactone (PCL) and polylactic acid (PLA) for cardiomyocyte differentiation of human Ad-MSC (hAd-MSCs) in the absence or presence of 5-azacytidine and transforming growth factor-ß (TGF-ß). To that end, the human MSCs were extracted from human adipose tissue (AD). The cardiomyocyte differentiation potency of hAd-MSCs was evaluated on the novel synthetic PCL/PLA nanofiber scaffolds prepared in the absence and presence of 5-azacytidine and TGF-ß supplements. A PCL/PLA nanofibrous scaffold was fabricated using the electrospinning method and its nanotopography and porous structure were characterized using scanning electron microscopy. In addition, the attachment of hAd-MSCs on the PCL/PLA scaffolds was semiquantitatively investigated. Compared with other treatments, the PCL/PLA nanofibrous scaffold supplemented with both 5-azacytidine and TGF-ß was observed to differentiate hAd-MSCs into cardiomyocytes at Day 21 as evidenced by real-time PCR for cardiac-specific genes including cardiac troponin I (cTnI), GATA4, MYH7, and NKX2.5. In addition, flow cytometric analysis of cTnI-positive cells demonstrated that the cardiomyocyte differentiation of hAd-MSCs was more efficient on the PCL/PLA nanofibrous scaffold supplemented with both 5-azacytidine and TGF-ß than it was in the other treatment groups. Generally speaking, the results show that PCL/PLA nanofibrous scaffolds may be applied as a platform for efficient differentiation of hAd-MSCs into functional cardiomyocytes.

Effects of bacteriophage supplement on the growth performance, microbial population, and PGC-1α and TLR4 gene expressions of broiler chickens.

Bacteriophages (BP) are viruses that invade bacteria and propagate inside them, leading to the lysis of the bacterial cells. The aim of this study was to investigate the effect of adding BP to the broiler's diet and its effect on the performance, morphology and bacterial population of the gut, some immune responses and expression of some intestinal genes. Accordingly, dietary treatments were as follows: basal diet (control), and control + 0.3 g/kg colistin or 0.5, 1 and 1.5 g BP/kg of diet. BP increased the body weight gain and reduced the feed conversion ratio (FCR), as compared to the colistin treatment, in the finisher and overall period (P < 0.05). European efficiency factor was significantly higher in 1.5 g BP-fed birds, as compared to the control and colistin treatments. meanwhile, bacteriophage and colistin-fed birds had higher Lactobacillus and lowered coliform bacteria counts, as compared to the control treatment (P < 0.05). Cecal concentrations of propionate in the 1.5 g BP-fed birds were higher than those in the control treatment (P < 0.05). BP-fed birds had a significantly increased villus height to crypt depth ratio, as compared to the control treatment. BP increased the serum concentrations of the total antibody, immunoglobulin (Ig) M, and IgG, as compared to the control treatment (P < 0.05). In the ileum, the expression of the Peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) gene was decreased by dietary BP supplementation (P < 0.05). Furthermore, Toll-like receptor 4 (TLR4) gene expression was down-regulated in the BP-fed birds, whereas Interleukin 10 (IL-10) gene expression was up-regulated (P < 0.05). Overall, the use of BP may be a promising alternative to growth-promoting antibiotics in broilers by altering the gastrointestinal tract microbiota, enhancing immunological responses and improving the gut's morphology.

Melatonin accelerates the developmental competence and telomere elongation in ovine SCNT embryos.

SCNT embryos suffer from poor developmental competence (both in vitro and in vivo) due to various defects such as oxidative stress, incomplete epigenetic reprogramming, and flaws in telomere rejuvenation. It is very promising to ameliorate all these defects in SCNT embryos by supplementing the culture medium with a single compound. It has been demonstrated that melatonin, as a multitasking molecule, can improve the development of SCNT embryos, but its function during ovine SCNT embryos is unclear. We observed that supplementation of embryonic culture medium with 10 nM melatonin for 7 days accelerated the rate of blastocyst formation in ovine SCNT embryos. In addition, the quality of blastocysts increased in the melatonin-treated group compared with the SCNT control groups in terms of ICM, TE, total cell number, and mRNA expression of NANOG. Mechanistic studies in this study revealed that the melatonin-treated group had significantly lower ROS level, apoptotic cell ratio, and mRNA expression of CASPASE-3 and BAX/BCL2 ratio. In addition, melatonin promotes mitochondrial membrane potential and autophagy status (higher number of LC3B dots). Our results indicate that melatonin decreased the global level of 5mC and increased the level of H3K9ac in the treated blastocyst group compared with the blastocysts in the control group. More importantly, we demonstrated for the first time that melatonin treatment promoted telomere elongation in ovine SCNT embryos. This result offers the possibility of better development of ovine SCNT embryos after implantation. We concluded that melatonin can accelerate the reprogramming of telomere length in sheep SCNT embryos, in addition to its various beneficial effects such as increasing antioxidant capacity, reducing DNA damage, and improving the quality of derived blastocysts, all of which led to a higher in vitro development rate.

Hepatic fatty acids profile, oxidative stability and egg quality traits ameliorated by supplementation of alternative lipid sources and milk thistle meal.

This study assessed the effects of different levels of milk thistle (Silybum marianum) meal (MTM) on egg quality traits, oxidative stability, blood biochemical parameters and liver health indices in laying hens receiving diets supplemented with lipid from different sources. For this purpose, Leghorn laying hens (Hy-Line W-36) were randomly assigned to 12 experimental treatments with eight replicates of five birds each. Dietary treatments consisted of four dietary fat sources (fish oil, sunflower oil, poultry oil and fat powder, each with different fatty acid profiles) and three levels of MTM (0.0, 15 and 30 g/kg) offered through 10 days of adaptation and 70 days of main recording. Results showed that dietary inclusion of polyunsaturated fatty acids, including fish or sunflower oils, reduced serum cholesterol content but increased blood malondialdehyde (MDA) concentration (p < 0.05). While fish oil improved the yolk colour index and Haugh unit (p < 0.05), it decreased yolk cholesterol concentration (p < 0.01). The highest hepatic glycogen content and tissue integrity as well as the lowest liver lipid percentage and MDA content were observed in birds fed diets supplemented with poultry oil. On the other hand, feeding 30 g/kg of MTM reduced not only hepatic lipid percentage but also aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activities (p < 0.01), while it improved eggshell strength and thickness (p < 0.05). Supplementation of fat powder enhanced hepatic C16:0 content, while addition of poultry, sunflower or fish oil increased C18:1, C18:2 or C20:5 concentrations respectively (p < 0.01); nonetheless, feeding the highest level of MTM reduced hepatic ΣSFA but increased Σn-3 fatty acids (p < 0.01). The interaction effects indicated that fish or sunflower oil supplementation increased the MTM level required to reduce not only serum or egg cholesterol concentrations (p < 0.05) but also blood or hepatic MDA content (p < 0.01). It was concluded that fish oil and MTM (30 g/kg) supplements might improve internal egg quality traits and eggshell quality respectively. The hepatic fatty acid profile was also found strongly correlated with dietary fatty acids. Finally, the best hepatic health indices and the highest oxidative stability were achieved when the birds were fed diets supplemented with poultry oil and 30 g/kg of MTM.

Role of Signaling Pathways during Cardiomyocyte Differentiation of Mesenchymal Stem Cells.

Multipotent stem cells, including mesenchymal stem cells (MSCs), represent a promising source to be used by regenerative medicine. They are capable of performing myogenic, chondrogenic, osteogenic, and adipogenic differentiation. Also, MSCs are characterized by the expression of multiple surface antigens, but none of them appear to be particularly expressed on MSCs. Moreover, the prospect of monitoring and controlling MSC differentiation is a scientifically crucial regulatory and clinical requirement. Different transcription factors and signaling pathways are involved in cardiomyocyte differentiation. Due to the paucity of studies exclusively focused on cardiomyocyte differentiation of MSCs, the present study aims at describing the roles of various signaling pathways (FGF, TGF, Wnt, and Notch) in cardiomyocyte differentiation of MSCs. Understanding the signaling pathways that control the commitment and differentiation of cardiomyocyte cells not only will expand our basic understanding of molecular mechanisms of heart development but also will enable us to develop therapeutic means of intervention in cardiovascular diseases.

Adipose-Derived Mesenchymal Stem Cells Differentiation Toward Cardiomyocyte-Like Cells on the PCL/PANI Nanofibrous Scaffold: An Experimental Study.

Background: Owing to the fact that the heart tissue is not able to repair itself. Biomaterial-based scaffolds are important cues in tissue engineering (TE) applications. Recent advances in TE have led to the development of suitable scaffold architecture for various tissue defects. Objective: Given the importance of cellular therapy, it was the aim of the present study to differentiate cardio myocyte cells from human adipose-derived mesenchymal stem cells (Ad-MSCs) using suitable induction reagents (namely, 5-azacytidine and transforming growth factor beta (TGF-ß)) on poly-caprolactone (PCL)/Poly aniline (PANI) Nano fibrous scaffolds prepared by electrospinning. Materials and Methods: For this purpose, the adipose-derived mesenchymal stem cells (Ad-MSCs) were initially isolated and characterized before cultivation on the PCL/PANI Nano fibrous scaffold to be treated for 21 days with 5-azacytidine either singly or in combination with TGF-ß in medium. The scaffold's morphological and cell attachment properties were investigated using electron microscopy (SEM). Finally, the cardio myocyte differentiation of Ad-MSCs on the scaffold was studied using both quantitative Real-time PCR (qPCR) and flow-cytometry while the expression rates of the cardio myocytes' specific genes (Gata4, NKX2.5, MYH-7, and Troponin I) were also determined. Results: The results of Ad-MSCs culture, MTT assay, and SEM indicated that the cells had well proliferated on the PCL/PANI scaffolds, showing the biocompatibility of the nanofibers for cellular growth and adhesion. After 21 days of induced cardio myocyte differentiation by both agents, Real-time PCR revealed increases in the expressions of Gata4, Troponin I, MYH-7, and NKX2.5 genes in the cells cultured on the PCL/PANI scaffolds while the flow-cytometry test approved the expression of troponin I. Conclusion: The data obtained showed that the PCL/PANI Nano fibrous scaffolds were able to promote and support mesenchymal stem cell transformation to cardio myocyte cells. Generally speaking, the results of the study might be exploited in future in vitro and in vivo experimental model studies of cardio myocyte differentiation using co-polymer scaffolds.

Sanguinarine improved nutrient digestibility, hepatic health indices and productive performance in laying hens fed low crude protein diets.

A major mean to minimize feeding costs and faecal nitrogen excretion on poultry farms is to decrease the supplied dietary protein content. This, however, is associated with the declines in productive performance and systemic health indices. Sanguinarine may improve protein efficiency via decreasing the intestinal amino acid decarboxylation and stimulating the tryptophan-serotonin pathway. The present study was carried out to investigate the effects of dietary supplementation of sanguinarine on the performance, egg yolk biochemical parameters, serum enzyme activities, nutrient digestibility, ovarian follicles, and hepatic health indices in laying hens fed decremental levels of crude protein (CP). For this purpose, 180 laying hens were allocated into nine dietary treatments with four replicates of five birds each. The experimental treatments consisted of three levels of CP (85.0%, 92.5%, and 100% of Hy-Line W-36 manual recommendation) and three levels of sanguinarine (0.00, 3.75, and 7.50 mg/kg) in a 3 × 3 factorial arrangement administered during a 70-day feeding trial. Results showed that the decremental levels of CP led to significant increases in serum aspartate aminotransferase (p < .05), alanine aminotransferase, and alkaline phosphatase (p < .01) activities, egg yolk cholesterol concentration (p = .064), and hepatic fat and malondialdehyde (MDA) contents (p < .05). It also caused the significant declines in ileal dry matter (DM) digestibility (p < .05) and eggshell strength (p < .05), and also tended to decrease CP digestibility (p = .071), Haugh unit (p = .057) and egg production percentage (p = .062). The interaction effects of the experimental factors indicated that dietary supplementation of sanguinarine, especially at 7.50 mg/kg, led to significant improvements in serum aspartate aminotransferase and alanine aminotransferase activities (p < .01), egg yolk cholesterol (p < .001) and triglyceride (p < .05) concentrations, eggshell strength (p < .001), Haugh unit (p < .05), hepatic fat (p < .001) and MDA (p = .059) contents, ileal DM and CP digestibility (p < .01) as well as egg production, egg mass and feed conversion ratio (FCR; p < .05) in birds receiving decremental levels of CP. Taken together, the results indicate that dietary administration of sanguinarine could enhance productive performance via improving nutrient digestibility, hepatic health indices and fortifying systemic antioxidant capacity in laying hens fed low-CP diets.

Impact of a Ration Negative in Dietary Cation-Anion Difference and Varying Calcium Supply Fed before Calving on Colostrum Quality of the Dams and Health Status and Growth Performance of the Calves.

This study investigated the effect of diets negative in dietary cation-anion difference (DCAD) or restricted in Ca fed prepartum to dairy cows for three weeks on colostrum yield and composition, and the health and growth performance of their calves. Thirty-six pregnant non-lactating Holstein-Friesian cows were randomly assigned to three isoenergetic diets: (1) low Ca: 0.24% Ca, DCAD: +86 mEq/kg; (2) high Ca: 1.23% Ca, DCAD: +95 mEq/kg; and (3) low DCAD: 1.28% Ca, DCAD: -115 mEq/kg (all dry matter (DM) basis). While colostrum quality was not affected, low Ca supply prepartum tended to increase the colostrum yield compared to high Ca (low Ca = 8.81 vs. high Ca = 5.39 kg). However, calves from cows fed low DCAD showed higher serum concentrations of K, lower body weight (BW), starter feed intake and average daily weight gain before weaning compared to low Ca and high Ca calves (53.12 vs. 57.68 and 57.32 kg) but BW was similar postweaning (d 70). In addition, calves from dams fed low DCAD were more likely to develop diarrhea and had increased number of days with abnormal fecal scores. Consequently, calves from low DCAD dams had to be treated more frequently.

Effects of Different Concentrations of Reversine on Plasticity of Mesenchymal Stem Cells.

Dedifferentiation can be induced by small molecules. One of these small molecules used in this study in order to increase the plasticity of differentiation of stem cells was reversine. The objective of present study was to investigate the effect of different concentrations of reversine on the plasticity of ovine fetal bone-marrow mesenchymal stem cells (BM-MSCs). BM-MSCs were extracted from ovine fetal and cultured. Passaged-3 cells were evaluated for their differentiation potential into osteocytes and adipocytes cells. In the present study, BM-MSCs were culture plated in the presence of 0, 300, 600, 900 and 1200 nM of reversine. The number of viable cells was determined by MTT test after addition of different concentrations of reversine. Furthermore, expression of the nanog gene was evaluated. The culture without reversine was taken as the control group. Expression of nanog was analysed by immunocytochemistry. Multi-lineage differentiation showed that the BM-MSCs could be differentiated into adipose cells and osteocytes. Our results indicated that the addition of 1200 nM of reversine to medium significantly decreased overall proliferation compared to the other treatment groups (p > 0.05). Real-time PCR analysis showed that after addition of 600 nM of reversine significantly increased nanog expression compared to other treatments. All treatments received reversine were seen to be relative expression of nanog. Our findings confirm that low concentrations reversine increases the plasticity of ovine BM-MSCs.

Positive effects of conjugated linoleic acid (CLA) on the PGC1-α expression under the inflammatory conditions induced by TNF-α in the C2C12 cell line.

The aim of the current study was to evaluate the effects of conjugated linoleic acid (CLA) on the expression of the genes involved in the reduction of inflammatory conditions induced by tumor necrosis factor alpha (TNF-α). To fulfill this, the C2C12 murine myoblast cell line was used. The experimental treatments consisted of 50 and 100 µM of CLA with 10, 15 and 20 ng/mL of TNF-α. Our results indicated that the concentrations of 50, 100 and 150 µM of CLA at 24 and 48 h increased the cell survival rate (p < 0.05), as compared with the control group. The effect of pre-treating CLA before the inflammation induced by TNF-α showed that the amount of cell death caused by TNF-α was inhibited in both 50 and 100 µM concentrations of CLA (p < 0.05). Modulatory effects of CLA on PGC1-α and Fndc5 transcript levels showed that the PGC1-α and Fndc5 expression levels were significantly increased 48 h after treating with 50 and 100 µM of CLA (p < 0.05). Overall, these findings indicate that using CLA could considerably improve the inflammatory condition induced by TNF-α.

The attenuation of the toxic effects of LPS on mouse pre-implantation development by alpha-lipoic acid.

The present study was conducted to evaluate the effects of alpha-lipoic acid (ALA) on quantitative and qualitative indices of mouse embryos challenged by lipopolysaccharide (LPS). Having determined the effective concentrations of LPS (1 mg/mL) that could reduce blastocyst formation rate by around 50% and the optimal concentration of ALA (10 µM) that could attenuate the toxic effects of LPS on blastocyst formation, the following indices were defined: inner cell mass and trophectoderm cell numbers, blastocyst mitochondrial distribution, ROS and GSH levels, as well as the relative expression of Tlr-4. Nrf-2 and Tnf-RI/P-60 receptor involved in inflammatory pathways. Finally, embryos derived from the experimental and control groups were transferred to synchronized recipients and their implantation rate and post-implantation capacity were determined. Treatment with LPS resulted in an increase in intracellular ROS level (P ≤ 0.05), and remarkable decreases (P ≤ 0.05) in intracellular GSH content, mitochondrial mass, and blastocyst quality. ALA attenuated all the aforementioned negative effects of LPS. The relative expression levels of Nrf-2 and Tnf-RI/P-60 receptor (P ≤ 0.05) significantly increased in response to LPS, and treatment with ALA significantly reduced the relative expression of Tnf-RI/P-60. ALA also improved the post-implantation developmental capacity of embryos treated with LPS. In conclusion, our findings indicate that the reproductive toxicity of LPS could be overcome by ALA treatment. These effects were mainly due to the improvements made in intracellular antioxidant capacity as well as suppression of some inflammatory elements, especially the main receptor of TNF-α, the Tnf-RI/P-60, involved in induction of apoptosis. These observations have important implications for dairy farming and treatment of infertility.

Cryopreservation of rooster semen: Evidence for the epigenetic modifications of thawed sperm.

Although semen cryopreservation is an important, widely used technique for long-term sperm storage, it not only induces partially irreversible damages to sperm but might also deteriorate anatomical, biochemical, and structural organelles. These cellular and epigenetic modifications are the main reasons underlying the decline in sperm motility and fertility during the freeze-thaw process. Using the two Lake and Beltsville semen extenders, the present study aims to evaluate the epigenetic patterns (DNA methylation and histone modification), cellular parameters (e.g., membrane integrity, viability, DNA stability, mitochondria activity, and apoptosis status), and fertility potential of rooster semen collected from six mature roosters before and after cryopreservation according to a standard protocol. The results show that cryopreservation leads to significantly (P < 0.05) reduced values of the parameters examined when compared with those of fresh sperms. While the extenders used exhibit no difference with respect to DNA methylation (DNMT), the Lake extender leads to significant reductions (P < 0.05) in H3K9 acetylation (17.4 ±â€¯1.8) and H3K4 methylation (42 ±â€¯2.3) compared to the Beltsville (9.2 ±â€¯1.8 and 23 ±â€¯2.3, respectively). Compared to the Beltsville extender, the Lake one is also observed to yield a significantly (P < 0.05) superior sperm quality in terms of total motility (TM; 77.2 ±â€¯1.6 vs. 68.3 ±â€¯1.6), average path velocity (VAP; 71 ±â€¯1.4 vs. 53 ±â€¯1.4), and straight-line velocity (VSL; 52 ±â€¯1.5 vs. 34 ±â€¯1.5) as well as significantly (P < 0.05) higher viability (60 ±â€¯1.69 vs. 51 ±â€¯1.69) and membrane functionality (55 ±â€¯3.2 vs. 46 ±â€¯3.2). The Lake extender is also found to outperform the Beltsville one due to its significantly (P < 0.05) higher fertility rate (59.5% vs. 47.2%). The two extenders, however, exhibit no differences in DNA fragmentation, mitochondrial activity, or hatchability rate. The Beltsville extender showed to be superior to the Lake one due to its significantly greater reactive oxygen species percentage (ROS; 45.9 ±.3.2 vs. 28.5 ±â€¯3.2) and apoptosis (29 ±â€¯2.3 vs. 27 ±â€¯2.3). It may be concluded that the Lake extender is capable of improving the cellular and epigenetic parameters of rooster sperms during cryopreservation due to the crucial roles it plays in the protection of sperms against cryo-damages.

Microwave spectroscopy based on the waveguide technique for the nondestructive freshness evaluation of egg.

A rectangular waveguide equipped with a network analyzer was used to assess the quality indices of shell egg. The scattering parameters of the eggs were acquired in the range of 0.9-1.7 GHz and they were then used to calculate microwave spectra of the samples. PLS and ANN regression methods were implemented to predict the egg quality indices and SIMCA and ANN classification methods were applied to classify the eggs based on their storage time. The best predictive models, however, obtained from ANN analysis where the yolk coefficient, air cell height, thick albumen height, Haugh unit, and albumen pH could be predicted with the residual predictive deviation (RPD) values of 3.500, 3.000, 2.411, 2.033, and 1.829, respectively. To classify the eggs according to their storage time, both SIMCA and ANN analyses resulted in the total accuracy of 100% when return loss spectra were used as the input.

Efficacy of essential oils combination on performance, ileal bacterial counts, intestinal histology and immunocompetence of laying hens fed alternative lipid sources.

This study was carried out to assess the effects of a blend of herbal essential oils (namely Thymus vulgaris, Mentha piperita, Rosmarinus offisinalis and Anethum graveolens) and different dietary lipid sources on the performance, ileal bacterial enumeration, intestinal histological alterations and immune responses in laying hens. For this purpose, a total of 150 laying hens were randomly allocated to six experimental treatments with five replicates of five birds each. Dietary treatments consisted of three levels of a mixture of essential oils (0, 100 and 200 mg/kg) and two sources of lipid (soybean oil and fish oil containing different ratios of n-6 to n-3 fatty acids) fed to the animals during an 80-days feeding trial. Findings indicated that dietary administration of fish oil not only increased significantly the spleen relative weight (p < 0.01) and the antibody titre against Newcastle virus (p < 0.05) but also led to reductions in liver relative weight (p < 0.05) and feed conversion ratio (p < 0.05). Moreover, the mixture of herbal essential oils brought about declines in hepatic relative weight, heterophile to lymphocyte ratio (p < 0.05) and intestinal pathogen populations (p < 0.01). Mention must also be made of the improvements it made in spleen weight (p < 0.01), antibody titres against SRBC (p < 0.01) and Newcastle virus (p < 0.05), villus height to crypt depth ratio (p < 0.01), goblet cell numbers (p < 0.05), lamina propria lymphatic follicle diameters (p < 0.01) and feed conversion ratio (p = 0.06). It may be claimed that the enhancements observed in the performance of laying hens fed fish oil and 200 mg/kg of the blend of essential oils could have potentially been associated with improved intestinal health indices as well as cellular and humoral immune responses.