Comparison of the differentiation of ovine fetal bone-marrow mesenchymal stem cells towards osteocytes on chitosan/alginate/CuO-NPs and chitosan/alginate/FeO-NPs scaffolds.

In the current study, the creation of a chitosan/alginate scaffold hydrogel with and without FeO-NPs or CuO-NPs was studied. From fetal ovine bone marrow mesenchymal stem cells (BM-MSCs) were isolated and cultivated. Their differentiation into osteocyte and adipose cells was investigated. Also, on the scaffolds, cytotoxicity and apoptosis were studied. To investigate the differentiation, treatment groups include: (1) BM-MSCs were plated in DMEM culture medium with high glucose containing 10% FBS and antibiotics (negative control); (2) BM-MSCs were plated in osteogenic differentiation medium (positive control); (3) positive control group + FeO-NPs, (4) positive control group + CuO-NPs; (5) BM-MSCs were plated in osteogenic differentiation medium on chitosan/alginate scaffold; (6) BM-MSCs were plated in osteogenic differentiation medium on chitosan/alginate/FeO-NPs scaffold; and (7) BM-MSCs were plated in osteogenic differentiation medium on chitosan/alginate/CuO-NPs scaffold. Alkaline phosphatase enzyme concentrations, mineralization rate using a calcium kit, and mineralization measurement by alizarin staining quantification were evaluated after 21 days of culture. In addition, qRT-PCR was used to assess the expression of the ALP, ColA, and Runx2 genes. When compared to other treatment groups, the addition of CuO-NPs in the chitosan/alginate hydrogel significantly increased the expression of the ColA and Runx2 genes (p < 0.05). However, there was no significant difference between the chitosan/alginate hydrogel groups containing FeO-NPs and CuO-NPs in the expression of the ALP gene. It appears that the addition of nanoparticles, in particular CuO-NPs, has made the chitosan/alginate scaffold more effective in supporting osteocyte differentiation.

Bupropion and varenicline administration reversed depressive behavior induced by acetamiprid exposure in mice: the role of nAChRs in depression.

Acetamiprid (ACE), is a popular neonicotinoid pesticide, that has a high affinity for mammalian nicotinic acetylcholine receptors (nAChRs). Therefore, ACE might induce depressive effects by perturbing the cholinergic system in mammalian. The aim of this study was to evaluate the effects of ACE exposure on depressive-like behaviors and grip strength (GS) in mice. Also the possible role of nAChR activation in depression was assessed by varenicline, and bupropion. Male Swiss mice (27 ± 2 g) were daily exposed to ACE by gavage (0.1, 1, 5 mg/kg), behavioral tests took place after 3 h, 7 days and 15 days, the subacute ACE (0.1 mg/kg) exposure was assessed after 30 days. Varenicline (0.5 mg/kg) or bupropion (4 mg/kg) were injected intraperitoneally 30 min prior exposure to (1 mg/kg) ACE. The locomotor activity, forced swimming test (FST), and sucrose preference (SP) test were assessed. After a week ACE dose dependently increased the immobility time during FST, and after 15 days' depressive behavior was observed equally for ACE (0.1-5 mg/kg). The subacute exposure (0.1 mg/kg) significantly increased the immobility time, SP also declined that revealed anhedonia. These behavioral changes showed that ACE can initiate depressive effects. The changes in locomotor activity were not significant. GS significantly reduced following a week of exposure to ACE (1-5 mg/kg) that indicated neurotoxicity. These effects were antagonized by bupropion or varenicline, thus ACE effect on nAChRs was essential in initiating the depressive behavior.

Effects of sodium selenite, cysteamine, bacterially synthesized Se-NPs, and cysteamine loaded on Se-NPs on ram sperm cryopreservation.

During the cryopreservation of sperm, the production of highly reactive oxygen species (ROS) can reduce their viability and fertility. However, the addition of antioxidants can help reduce the harmful effects of ROS. One such antioxidant is selenium, which is a co-factor of the glutathione peroxidase enzyme that is effective in scavenging ROS. Cysteamine can also take part in the structure of this enzyme. The use of nanoparticles can be less toxic to cells than their salt form. To this end, researchers synthesized Se-NPs using the streptococcus bacteria and loaded cysteamine onto the synthesized Se-NPs. The biosynthesis of Se-NPs and cysteamine loaded on Se-NPs was confirmed by UV-visible spectroscopy, X-ray diffraction (EDX), Fourier transforms infrared (FTIR) spectroscopy, and Field Emission Scanning Electron Microscope (FE-SEM). For cryopreservation, ram semen samples were diluted, and different concentrations (0, 1, 5, 25, and 125 µg/mL) of cysteamine, Se-NPs, cysteamine loaded on Se-NPs, and sodium selenite were added. An extender containing no supplement was considered as control group. After cooling the semen samples, they were frozen and stored in liquid nitrogen for evaluation. The samples were thawed and analyzed for mobility, viability, membrane and DNA integrity, and sperm abnormalities, as well as malondialdehyde level (MDA) and superoxide dismutase (SOD). The data was processed using SPSS, and a significance level of p < 0.05 was considered. The results of this experiment showed that adding 1 µg/mL of cysteamine loaded on Se-NPs to the diluent significantly increased the motility, viability, and membrane integrity and SOD of spermatozoa compared to the other treatment groups and control group, and reduced the abnormality, apoptosis, and MDA level of spermatozoa in comparison with the other treatment groups and control group (p < 0.05). In conclusion, the addition of cysteamine loaded on Se-NPs was found to improve the quality of ram sperm after cryopreservation.

Chitosan/alginate scaffold enhanced with Berberis vulgaris extract for osteocyte differentiation of ovine fetal stem cells.

Designing biocompatible polymers using plant derivatives can be extremely useful in tissue engineering, nanomedicine, and many other fields of medicine. In this study, it was first looked into how chitosan/alginate scaffolds were made and characterized in the presence of berberine and barberry fruit extract. Second, the process of proliferation and differentiation of ovine fetal BM-MSCs (bone marrow-mesenchymal stem cells) was assessed on these scaffolds after BM-MSCs were extracted and confirmed by developing into osteocyte and adipose cells. To investigate the differentiation, treatment groups include (1) ovine fetal BM-MSCs were plated in Dulbecco's modified eagle medium culture medium with high glucose containing 10% fetal bovine serum and antibiotics (negative control), (2) ovine fetal BM-MSCs were plated in osteogenic differentiation medium (positive control group), (3) positive control group + barberry fruit extract, (4) positive control group + berberine, (5) ovine fetal BM-MSCs were plated in osteogenic differentiation medium on chitosan/alginate scaffold (hydrogel group), (6) ovine fetal BM-MSCs were plated in osteogenic differentiation medium on chitosan/alginate/barberry fruit extract scaffold (hydrogel group containing barberry fruit extract), and (7) ovine fetal BM-MSCs were plated in osteogenic differentiation medium on chitosan/alginate/berberine scaffold (hydrogel group containing berberine). Alkaline phosphatase (ALP) enzyme concentrations, mineralization rate using a calcium kit, and mineralization measurement by alizarin staining quantification were all found after 21 days of culture. In addition, real-time quantitative reverse transcription polymerase chain reaction was used to assess the expression of the ALP, COL1A2, and Runx2 genes. Days 5 and 7 had the lowest water absorption by the hydrogel scaffold containing barberry extract, which was significant in comparison to other groups (p < .05). Among the hydrogel scaffolds under study, the one containing barberry extract exhibited the lowest tensile strength, and this difference was statistically significant (p < .05). The chitosan/alginate hydrogel has the highest tensile strength of all of them. In comparison to the control and other treatment groups, the inclusion of berberine in the chitosan/alginate hydrogel significantly increased the expression of the ALP, Runx2, and COL1A2 genes (p < .05). The osteocyte differentiation of mesenchymal stem cells in in vitro settings appears to have been enhanced by the inclusion of berberine in the chitosan/alginate scaffold.

DNA interaction studies of a cobalt(III) complex containing ß-amino alcohol ligand by spectroscopic and molecular docking methods.

In the present research, the feasibility of a Cobalt(III) complex containing ß-amino alcohol ligands for affinity with the target calf thymus DNA is demonstrated. In the title complex, [Co(C11H15N2O2)2]Cl, the Co(III) atom is six-coordinated with four N atoms and two O atoms from (2-[(E)-({2-[(2-Hydroxyethyl) amino]ethyl}imino)methyl]phenol) ligand (L). To investigate the molecular interaction between the synthesized complex and DNA, some multi-spectroscopic approaches associated with molecular docking were employed in the physiological buffer (pH 7.4). The results indicated that the Co(III) complex proved to be a minor groove binder with a preference for the A-T region, which was substantiated by displacement studies with Hoechst33258 and Methylene blue (MB) as minor groove binder and intercalator. In addition, the results of the molecular docking study revealed that the Co(III) complex approached the gap between the DNA minor grooves near the spot where the Hoechst was. Furthermore, the results of the cytotoxicity and apoptosis tests for the MCF-7 cell line were also indicative of the positive effects of the complex on controlling the growth and viability of breast cancer.Communicated by Ramaswamy H. Sarma.

Evaluation of DNA damage induced by acesulfame potassium: spectroscopic, molecular modeling simulations and toxicity studies.

Acesulfame potassium (Ace-K) is a widely used artificial sweetener that has been reported to interact with DNA and cause important genetic damage. However, the type of interaction mechanism is unknown. This study provides an approach to understanding the in vitro mechanism of Ace-K interaction with Ct-DNA using spectroscopic methods combined with molecular simulations. The hypochromic effect as obtained from UV-Vis spectra indicated the formation of the DNA-Ace-K complex in the minor groove. Further evidence for groove binding mode comes from the decrease in Hoechst-DNA fluorescence caused by increasing Ace-K concentrations, alongside no detectable change in MB-DNA emission band intensity. A negative value of ΔH and ΔS represents the hydrogen bonds and van der Waals forces between Ace-K and DNA. Based on the molecular docking, Ace-K was located between the guanine10 and 16 in DNA minor groove and stabilized by two hydrogen bonds and one π-Sulfur interaction. In vitro cell culture results showed that about 5 mg/mL of Ace-K caused the death of 85% of HUVEC cells after 48 h. Communicated by Ramaswamy H. Sarma.

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.

Effect of different concentrations of melatonin on ram epididymal spermatozoa recovered post-mortem under oxidative stress conditions and storage at 4°C.

The current study examines the protective effects of different melatonin concentrations on fresh ram epididymis spermatozoa after post-mortem recovery under normal and oxidative stress conditions and during liquid preservation (4°C) at different times (24, 48 and 72 h). The testes were obtained from a local slaughterhouse during the breeding season. Spermatozoa were isolated from cauda epididymides. In experiment 1, the effects of adding different concentrations of melatonin (0, 15, 60 and 240 µg/ml) under normal and oxidative stress conditions were evaluated. Fifty µM of hydrogen peroxide was used to induce oxidative stress. Also, in experiment 2, the spermatozoa samples were chilled to 4°C and stored for 72 h. Sperm total motility, viability, membrane, DNA integrity and morphological abnormality were evaluated at 0, 24, 48 and 72 h after cooling storage. In experiment 1, melatonin treatment preserved viability increased TAC and SOD activities, and reduced MDA levels compared with control. Also, melatonin reduced the harmful effects of H2 O2 under induced oxidative stress. In experiment 2, melatonin at concentrations of 15 and 60 g/ml greatly increased sperm viability after 3 days of cold storage. Furthermore, it could have a significant protective effect on the motility of cooled sperm. Melatonin supplementation preserved higher sperm membrane integrity at concentrations of 15 and 60 µg/ml, DNA integrity at a concentration of 15 µg/ml and abnormality at a concentration of 60 µg/ml after 3 days of storage. The results suggest that melatonin can be used to reduce the adverse effects of induced oxidative stress in spermatozoa. Furthermore, ram epididymal spermatozoa could be stored at 4°C for 72 h when treated with melatonin.

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 different concentrations of zinc oxide nanoparticles on the quality of ram cauda epididymal spermatozoa during storage at 4°C.

Zinc oxide nanoparticles (ZnO-NPs) are known for their antioxidant effects. In this study, ZnO-NPs were synthesized using aqueous extract of Maclura pomifera fruit; then, their effects on the quality of ram cauda epididymal spermatozoa were evaluated during storage at 4°C. ZnO-NPs formation was characterized by ultraviolet-visible spectroscopy (UV-vis), energy dispersive spectroscopy (EDX), field emission scanning electron microscopes (FE-SEM) and Fourier transform infrared (FTIR) spectroscopy. Cytotoxicity responses were investigated by 3-[4,5-dimethylthiazole-2-yl]-2,5-diphenyltetrazolium bromide (MTT). Cauda epididymal spermatozoa were obtained from testicles collected from abattoir(s). The sperm samples were pooled. The samples were diluted by extender and supplemented with different concentrations of ZnO-NPs (0, 1, 5 and 25 µg/ml) and cooled to 4°C for 72 hr. Total motility, viability, DNA and membrane integrity, morphological abnormality, malondialdehyde (MDA) and antioxidant activities: superoxide dismutase (SOD) and total antioxidant capacity (TAC) of cooled diluted samples were evaluated. Addition of 1 µg/ml of ZnO-NPs increased sperm viability under normal conditions (p < .05). Extender supplementation with 1 µg/ml of ZnO-NPs improved sperm total motility, viability, DNA and membrane integrity during storage periods (p < .05). Moreover, using 1 µg/ml of ZnO-NPs improved (p < .05) MDA, TAC and SOD activities after 72 hr compared to other treatments. In conclusion, there were some beneficial effects of ZnO-NPs supplementation in ram epididymal sperm extender during oxidative stress conditions and cooled storage.

Pre-treatment of ram semen extender with magnetic nanoparticles on freeze-thawed spermatozoa.

BACKGROUND: Extensive use of different nanoparticles caused significant concerns about their biological safety. OBJECTIVE: This study aimed to evaluate the effects of cryopreservation on ram semen after adding magnetic nanoparticles (MNPs) to separate X and Y chromosome-bearing spermatozoa. METHODS: The experimental ram sperms in this research included treated spermatozoa (50 µg/ml MNPs) and non-treated spermatozoa. DNA damage of spermatozoa was examined using an acridine orange (AO) assay. Sperm viability, membrane functionality, abnormality and malondialdehyde (MDA) level were also measured. RESULTS: Results indicated that the pre-treatment of ram semen extender with MNPs did not significantly affect the semen parameters such as viability, membrane functionality, abnormality, as well as lipid peroxidation (LPO) levels and DNA integrity in comparison with the control group (p < 0.05). CONCLUSIONS: These observations suggest that pre-treatment of ram semen extender with MNPs after semen sexing did not have adverse effects on different semen parameters after cryopreservation.

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.

Anti-proliferative, apoptotic potential of synthesized selenium nanoparticles against breast cancer cell line (MCF7).

Nano-biotechnology has grown rapidly and become an integral part of modern disease diagnosis and treatment. The aim of this survey was to evaluate the anticancer activity of synthesized selenium nanoparticles (Se-NPs) against breast cancer cells (MCF-7). The prepared Se-NPs were examined by ultraviolet-visible spectroscopy, Fourier transform infrared (FTIR) spectroscopy, scanning electron microscope (SEM), and energy dispersive spectroscopy (EDX). Antioxidant activity of Se-NPs property was studied by radical scavenging (DPPH) assay. The in-vitro cytotoxicity of Se-NPs was evaluated by MTT assay. In addition; the biological assessment (antioxidant and cytotoxicity) of synthesized Se-NPs was examined via molecular docking simulations. Synthesis of Se-NPs was characterized by several studies such as UV-absorbance, showing peak values in the range of 268 nm. Nanoparticle sizes of the nanoparticles are confirmed by dynamic light scattering analysis, indicating that average size is about 203 nm. The quantity of selenium in Se-NPs is 90.15% by weight, as confirmed by EDX. Synthesized Se-NPs have anti-proliferative effects on MCF-7 cell lines. Cytotoxicity and apoptotic potential assays exhibited a dose-dependent effect against MCF-7 cells using an MTT assay. Like anti-cancer activity, anti-oxidant activity of Se-NPs was dose-dependent. Findings showed that the Se-NPs complexes have the highest inhibitory effect against cytotoxic and antioxidant receptors. Results of this study demonstrated that Se-NPs had strong potential to scavenge free radicals and are cytotoxic against the MCF-7 cancer cell line.

Effects of hydroethanolic extracts of Terminalia chebula and Thymbra spicata on ram fresh semen under normal and oxidative stress conditions.

The objective of this study was to evaluate protective effects of hydroethanolic extracts of Terminalia chebula and Thymbra spicata on viability, lipid peroxidation (LPO) and DNA integrity of ram fresh semen under normal and oxidative stress (OS) conditions. Antioxidant activities of different concentrations of Terminalia chebula and Thymbra spicata extracts were evaluated with DPPH assay. Semen samples were taken from three fertile adult rams. After diluting semen with Tris-base extender, different concentrations of Terminalia chebula and Thymbra spicata (30, 300, and 3000 µg/ml) extracts were used under normal and induced OS conditions. The group not receiving any supplements was considered as control group. A total of 50 µM hydrogen peroxide was used to induce OS. MTT solution was added to each of treatment groups which were kept in an incubator at 37°C for 2 h. After incubation, readings were obtained by ELISA reader. DNA integrity and LPO were determined with acridine orange (AO) staining and malondialdehyde (MDA) assay. Higher concentrations of Terminalia chebula and Thymbra spicata extracts preserved viability and DNA integrity while reducing MDA concentrations compared to other treatment groups. Also, under induced OS, higher concentrations of both extracts reduced detrimental effects of H2 O2 . In conclusion, it seems that addition of Terminalia chebula and Thymbra spicata extracts can reduce induced OS in spermatozoa.

Effects of different concentrations of Chir98014 as an activator of Wnt/beta-catenin signaling pathway on oocyte in-vitro maturation and subsequent embryonic development in Sanjabi ewes.

The present study was conducted to investigate the effects of the activator factor of the WNT pathway, chir98014, leading to the in vitro sheep oocyte maturation medium, on the cumulus cell development, different nuclear maturation stages and the following process of embryonic development. Experiments included (a) addition of different concentrations (0, 0.1, 0.5, 1 µm) of chir98014 to the maturation medium and evaluation of the cumulus cell expansion, (b) addition of different concentrations of chir98014 to the maturation medium and investigation of different nuclear maturation stages, (c) addition of different concentrations of chir98014 to the maturation medium and examination of the subsequent embryonic maturation process and (d) addition of different concentrations of chir98014 to the embryonic development culture medium (the first 48 hr) and investigation of the subsequent embryonic development process. The extracted data were analysed using the SPSS software, considering the significance level of p < .05 and making the mean comparisons. The results showed that the addition of the 0.1 µM concentration of chir98014 to the maturation medium had no significant effects on the oocyte maturation and embryo development post-fertilization but it enhanced the Cumulus-oocyte complexes (COCs) expansion. In the fourth experiment, the low concentration of chir98014 in the embryo culture media improved the embryo development process, whereas the high one had a detrimental effect on it, as compared to the control group. Thus, the presence of the lower concentrations of this compound in the embryonic culture medium had favourable effects on the development of embryos.

Recovery of sperms bearing X chromosomes with different concentrations of magnetic nanoparticles in ram.

Pre-conceptual sex selection is still a highly debatable process whereby X and Y chromosome bearing spermatozoa are isolated before oocyte fertilization. Recently, magnetic nanoparticles (MNP) have been used to determine X and Y chromosomes bearing spermatozoa as a result of searching for a cheap, highly efficient method using non-toxic materials. This study aimed to recover the sperm bearing X chromosomes in ram with different concentrations of MNP and then evaluate the success of this method using polymerase chain reaction (PCR). Ram sperms were divided into four groups, treated with 0 (control), 50, 100 and 200 µg/ml MNP, respectively. MNP was used to restore sperm cells bearing X chromosomes. Upon recovery, the PCR was performed to identify the X and Y sperms, Methyl ThiazoleTetrazolium (MTT), to assess MNP toxicity and sperm viability and acridine orange (AO) to evaluate sperm DNA integrity. The results of PCR revealed that the treatment of spermatozoa- bearing X chromosomes with 50 µg/ml MNP had the highest effects on the recovery of X sperm rather than the other concentrations of MNP. However, the concentrations of MNP did not have any toxic effects on spermatozoa, sperm viability and, DNA integrity, but the high concentration of MNP (200 µg/ml) significantly reduced DNA integrity. According to MTT and AO results, the concentrations of MNP used in this study had no toxic effects on spermatozoa and did not reduce the sperm viability and DNA integrity, except that 200 µg/ml MNP significantly reduced DNA integrity.

Elevated serum levels of S100A1 and zinc α2-glycoprotein in patients with heart failure.

BACKGROUND AND AIMS: Heart failure (HF) is a growing concern worldwide. S100A1 and zinc α2-glycoprotein (ZAG) play an important role in heart function. We examined serum levels of S100A1 and ZAG in HF patients and their association with anthropometric indices and body composition. METHODS AND RESULTS: Sixty-four patients with HF, mean age 56.2, 48 male and 16 females, with ejection fraction <30-35%, were recruited from Shahid Madani Heart Hospital in Tabriz, Iran, from April to October 2019. Two groups, cachexia (n = 32) and non-cachexia (n = 32), which were divided based on weight loss of at least 7.5% in the last six months, were compared with the control group (n = 26). S100A1 and ZAG serum levels were determined by ELISA. Serum median (min-max) levels of S100A1 and ZAG were significantly greater in HF patients [326 (184.8-635.2) and 150.4 (61.5-520.7)] than healthy controls [265.4 (43.6-658.8) and 119.8 (16.7-533)], both p = 0.001. S100A1 Serum levels in cachexia group was significantly higher than non-cachexia group [331 (245.6-469.6) vs. 318 (184.8-635.2), p = 0.03]. A strong positive association was observed between S100A1 and ZAG serum levels in patients (r = 0.70, p < 0.0001). Serum levels of these two proteins negatively and significantly associated with BMI (r = -0.25, p = 0.044 and r = -0.28, p = 0.024, respectively) and arm circumference (r = -0.26, p = 0.037 and r = -0.25, p = 0.047, respectively). CONCLUSION: The results indicate that S100A1 and ZAG are likely to contribute to the pathogenesis of HF disease and weight loss, as well as the strong association between S100A1 and ZAG possibly indicating a similar mechanism of action for these two proteins.

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.