Identification of proteins associated with cell repair from canine adipose derived mesenchymal stem cells isolated conditioned media

Ahmad Yar Qamar,Xun Fang,Jongki Cho 한국수정란이식학회 2018 한국수정란이식학회 학술대회 Vol.2018 No.11

The objective of this study was to identify the proteins actively involved in the protection and repair of damaged cells, secreted by canine adipose derived mesenchymal stem cells (AT-MSCs) into the conditioned media. For this purpose, conditioned media (CM) was recovered from passage three stage canine AT-MSCs and skin fibroblasts cultured in serum free media after 24, 48 and 72 h. The extraction of exosomes was performed from 10-20 ml of CM using total exosome isolation kit. The isolated exosomes were then subjected to western analysis for the identification of annexin-I, annexin-II, histone H3 and dysferlin proteins. Results demonstrated the expression of proteins in the conditioned media isolated from canine AT-MSCs reflecting their potential in reducing the extent of damage at cellular levels. In conclusion, the conditioned media derived from canine AT-MSCs can be helpful in restoring the normal structure of cells both in vivo and in vitro conditions.

Role of antioxidants in fertility preservation of sperm - A narrative review

Ahmad Yar Qamar,Muhammad Ilyas Naveed,Sanan Raza,Xun Fang,Pantu Kumar Roy,Seonggyu Bang,Bereket Molla Tanga,Islam M. Saadeldin,Sanghoon Lee,Jongki Cho Asian Australasian Association of Animal Productio 2023 Animal Bioscience Vol.36 No.3

Male fertility is affected by multiple endogenous stressors, including reactive oxygen species (ROS), which greatly deteriorate the fertility. However, physiological levels of ROS are required by sperm for the proper accomplishment of different cellular functions including proliferation, maturation, capacitation, acrosomal reaction, and fertilization. Excessive ROS production creates an imbalance between ROS production and neutralization resulting in oxidative stress (OS). OS causes male infertility by impairing sperm functions including reduced motility, deoxyribonucleic acid damage, morphological defects, and enhanced apoptosis. Several in-vivo and in-vitro studies have reported improvement in quality-related parameters of sperm following the use of different natural and synthetic antioxidants. In this review, we focus on the causes of OS, ROS production sources, mechanisms responsible for sperm damage, and the role of antioxidants in preserving sperm fertility.

Improved Preimplantation Development of Porcine Cloned Embryos by Flavone Supplement as Antioxidant

FANG XUN,Ahmad Yar Qamar,윤기영,조종기 사단법인 한국동물생명공학회 2018 한국동물생명공학회지 Vol.33 No.4

This experiment was conducted to analyse the effects of flavone supplementation on the preimplantation development of in-vitro produced porcine embryos. During in-vitro development, immature oocytes and early embryos were exposed to different concentrations of flavone (0, 1μM, 25μM, 50 μM, and 100 μM respectively). Results showed that 100 μM of flavone significantly reduced the intracellular ROS levels of oocytes accompanied with a significant rise in GSH level. In parthenogenesis, no significant change was observed in the cleavage rates whether flavone was supplemented in IVM or IVC media. In IVM supplemented group, the blastocyst development rate was significantly enhanced by 1 μM concentration than other groups (51.5% vs. 41.3%, 44.0%, 36.3%, 31.7%; P<0.05) respectively. However, in IVC group 1 μM concentration significantly improved the blastocysts production than 50 μM and control groups (50.0% vs. 40.5%, 38.0%; P<0.05) respectively. Following nuclear transfer, the cleavage rate of IVM group was significantly more in 1 μM than 50 μM and 100 μM groups (92.9% vs. 89.7%, 87.8%; P<0.05), followed by similar pattern of cloned blastocysts production being significantly higher in 1 μM group than 50 μM, 100 μM and control groups (16.8% vs. 9.0%, 7.1%, 12.8%; P<0.05) respectively. In IVC group, 1 μM concentration resulted in significantly higher cleavage rate than 25 μM and 50 μM groups (91.7% vs. 87.8%, 88.8%; P<0.05) respectively. However, the blastocysts production was significantly higher in 100 μM group than others (26.2% vs. 13.6%, 14.0%, 18.2%; P<0.05) respectively. The optimal concentrations of flavone significantly enhanced the percentages of ICM:TE than control group (43.8% vs. 37.6%; P<0.05) accompanied with significantly higher expression levels of reprogramming related genes. In conclusion, the optimal concentrations of 1 μM during IVM and 100 μM during IVC can significantly improve the production of porcine in-vitro embryos.

Improved Preimplantation Development of Cloned Porcine Embryos through Supplementation of Histone Deacetylase Inhibitor MS-275

Xun Fang,Ahmad Yar Qamar,신상태,조종기 한국임상수의학회 2019 한국임상수의학회지 Vol.36 No.5

The objective of this study was to analyse the effects of MS-275 (Class I and II histone deacetylase inhibitor) supplementation on the development of porcine in-vitro somatic nuclear transfer embryo production. During in-vitro development, early embryos were exposed to different concentrations of MS-275 (0, 5 μM, 10 μM, and 20 μM). In in-vitro culture supplemented group, the blastocyst development rate was significantly enhanced by 10 μM concentration than other groups (24.0% vs. 19.3%, 21.8%, 11.5%; P < 0.05). Additionally, the 6 h supplementation group, significantly improved the blastocysts production than 24 h, 48 h and control groups (26.1% vs. 17.0%, 15.2%, 2.8%; P < 0.05). Following supplementation with optimal concentrations and time (10 μM-6 h group), the blastocyst production was significantly higher than control (25.7% vs 15.8%; P < 0.05). The optimal concentrations of MS-275 significantly enhanced the percentages of ICM:TE than control (43.6% vs. 38.4%; P < 0.05) accompanied with significantly higher expression levels of reprogramming related genes (POU5F1, Naong, and SOX2). In conclusion, the optimal concentrations of 10 μM MS-275 and 6 h supplementation during in-vitro culture can significantly improve the quality of porcine in-vitro somatic nuclear transfer embryos through histone acetylation and epigenetic modification. Increasing the efficiency of clonal animal production will greatly promote the development of animal disease models and xenotransplantation.

Perosomus elumbus fetal monster: a rare cause of dystocia in a beetal goat - a case report from Pakistan

Mubbashar Hassan,Sanan Raza,Ahmad Yar Qamar,Naveed Muhammad Ilyas,Mateen Abdul,Noman Muhammad,Sayed Aun Muhammad,Abid Hussain Shahzad 사단법인 한국동물생명공학회 2024 Journal of Animal Reproduction and Biotechnology Vol.39 No.1

Dystocia, a challenging condition in obstetrics, can arise from various causes, including fetal monsters with structural abnormalities. This case report presents a unique case of dystocia due to a fetal monster known as Perosomus Elumbis in a beetal breed goat from Pakistan. The 4-years-old pregnant doe presented with prolonged straining and failure to deliver the fetus after 8 hours of labor. Upon examination, the cervix was dilated, and only the forelimbs of the fetus were visible in the birth canal. The subsequent delivery involved the application of manual traction by using a dystocia kit, and the removal of edematous fluid from the legs. The monster fetus exhibited absence of hair growth, along with the absence of thoracic vertebrae. Two other fetuses were present, with one found dead and the other alive. Posttreatment involved fluid therapy, antibiotics, and supportive care for the doe. This case report sheds light on the occurrence of Perosomus Elumbis fetal monsters and their impact on dystocia in goat breeding. Understanding the underlying causes and implementing appropriate management strategies are crucial for successful outcomes in similar cases.

Canine amniotic membrane derived mesenchymal stem cells exosomes addition in canine sperm freezing medium

Mahiddine, Feriel Yasmine,Qamar, Ahmad Yar,Kim, Min Jung The Korean Society of Animal Reproduction and Biot 2020 한국동물생명공학회지 Vol.35 No.3

Amniotic membrane stem cells are considered as a good alternative to embryonic stem cells, but their use in clinical studies is still not common. Here, exosomes from canine amniotic membrane mesenchymal stem cells (cAmMSC-exo) were used for dog sperm cryopreservation. Upon cryopreserved straws using cryoprotectant containing 0, 0.5, 1, or 2 ㎍/mL of cAmMSC-exo were thawed, motility and membrane integrity were analyzed. However, results showed no significant differences between the groups. We concluded that cAmMSC-exo with lower than 2 ㎍/mL have no effects on sperm cryopreservation, and further studies to get higher concentrations of cAmMSC-exo should be conducted for clinical application.

Canine amniotic membrane derived mesenchymal stem cells exosomes addition in canine sperm freezing medium

Feriel Yasmine Mahiddine,Ahmad Yar Qamar,Min Jung Kim 한국동물생명공학회(구 한국동물번식학회) 2020 Journal of Animal Reproduction and Biotechnology Vol.35 No.3

Amniotic membrane stem cells are considered as a good alternative to embryonic stem cells, but their use in clinical studies is still not common. Here, exosomes from canine amniotic membrane mesenchymal stem cells (cAmMSCexo) were used for dog sperm cryopreservation. Upon cryopreserved straws using cryoprotectant containing 0, 0.5, 1, or 2 μg/mL of cAmMSC-exo were thawed, motility and membrane integrity were analyzed. However, results showed no significant differences between the groups. We concluded that cAmMSC-exo with lower than 2 µg/mL have no effects on sperm cryopreservation, and further studies to get higher concentrations of cAmMSC-exo should be conducted for clinical application.

Semen evaluation: methodological advancements in sperm quality-specific fertility assessment - A review

Tanga, Bereket Molla,Qamar, Ahmad Yar,Raza, Sanan,Bang, Seonggyu,Fang, Xun,Yoon, Kiyoung,Cho, Jongki Asian Australasian Association of Animal Productio 2021 Animal Bioscience Vol.34 No.8

Assessment of male fertility is based on the evaluation of sperm. Semen evaluation measures various sperm quality parameters as fertility indicators. However, semen evaluation has limitations, and it requires the advancement and application of strict quality control methods to interpret the results. This article reviews the recent advances in evaluating various sperm-specific quality characteristics and methodologies, with the help of different assays to assess sperm-fertility status. Sperm evaluation methods that include conventional microscopic methods, computer-assisted sperm analyzers (CASA), and flow cytometric analysis, provide precise information related to sperm morphology and function. Moreover, profiling fertility-related biomarkers in sperm or seminal plasma can be helpful in predicting fertility. Identification of different sperm proteins and diagnosis of DNA damage has positively contributed to the existing pool of knowledge about sperm physiology and molecular anomalies associated with different infertility issues in males. Advances in methods and sperm-specific evaluation has subsequently resulted in a better understanding of sperm biology that has improved the diagnosis and clinical management of male factor infertility. Accurate sperm evaluation is of paramount importance in the application of artificial insemination and assisted reproductive technology. However, no single test can precisely determine fertility; the selection of an appropriate test or a set of tests and parameters is required to accurately determine the fertility of specific animal species. Therefore, a need to further calibrate the CASA and advance the gene expression tests is recommended for faster and field-level applications.

Perosomus elumbus fetal monster: a rare cause of dystocia in a beetal goat - a case report from Pakistan

Mubbashar Hassan,Sanan Raza,Ahmad Yar Qamar,Muhammad Ilyas Naveed,Abdul Mateen,Muhammad Noman,Sayed Aun Muhammad,Abid Hussain Shahzad The Korean Society of Animal Reproduction and Biot 2024 한국동물생명공학회지 Vol.39 No.1

Dystocia, a challenging condition in obstetrics, can arise from various causes, including fetal monsters with structural abnormalities. This case report presents a unique case of dystocia due to a fetal monster known as Perosomus Elumbis in a beetal breed goat from Pakistan. The 4-years-old pregnant doe presented with prolonged straining and failure to deliver the fetus after 8 hours of labor. Upon examination, the cervix was dilated, and only the forelimbs of the fetus were visible in the birth canal. The subsequent delivery involved the application of manual traction by using a dystocia kit, and the removal of edematous fluid from the legs. The monster fetus exhibited absence of hair growth, along with the absence of thoracic vertebrae. Two other fetuses were present, with one found dead and the other alive. Posttreatment involved fluid therapy, antibiotics, and supportive care for the doe. This case report sheds light on the occurrence of Perosomus Elumbis fetal monsters and their impact on dystocia in goat breeding. Understanding the underlying causes and implementing appropriate management strategies are crucial for successful outcomes in similar cases.

Supplementation of cryoprotective extender with resveratrol decreases apoptosis index and reactive oxygen species levels in post-thaw dog sperm

Bang, Seonggyu,Tanga, Bereket Molla,Qamar, Ahmad Yar,Fang, Xun,Seong, Gyeonghwan,Nabeel, Abdelbagi Hamad Talha,Yu, Iljeoung,Cho, Jongki The Korean Society of Veterinary Science 2021 大韓獸醫學會誌 Vol.61 No.4

Resveratrol (RSV, 3,5,4'-trihydroxytrans-stilbene) protects sperm from cryo-induced damage in various animal and human species. In this study, we aimed to assess the effect of dog sperm cryoprotective extender containing RSV on the quality of post-thaw dog sperm. Sperm were collected from 4 Beagles and supplemented with different concentrations of RSV (0, 100, 200, and 400 µM). After thawing, apoptosis index, and reactive oxygen species (ROS) levels were assessed to determine post-thaw sperm quality. Dog sperm cryopreserved with 400 µM RSV showed significant improvement in post-thaw sperm quality with lower apoptosis index and ROS levels (p < 0.05). Our results showed that the supplementation of dog sperm cryoprotective extender with RSV at a concentration of 400 µM improved the post-thaw dog sperm quality in the term of sperm ROS production and apoptosis. In addition, we emphasize the necessity of testing the ROS levels and apoptosis index using flow cytometry to determine the quality of post-thaw semen.