The effects of vitamin E supplementation on sperm parameters, chromatin integrity, and gene expression before and after freezing in aged mice.

Objective: Some age-related testicular changes, such as Sertoli cell vacuolization and blood-testis barrier breakdown, reduce total sperm production and male fertility. Therefore, this study investigated the effect of vitamin E on restoring testicular function in aged mice. Sperm cryo-resistance was also assessed. Methods: Twenty-eight 48-week-old male Naval Medical Research Institute mice were divided into four groups for a daily gavage of vitamin E: the control group received distilled water, while the three treatment groups were administered 100, 200, and 400 mg/kg, respectively, for 4 weeks. Subsequently, semen analyses, DNA fragmentation index (DFI), and protamine deficiency tests were conducted. Testicular histology, tissue antioxidant enzyme activity, and gene expression levels were also assessed. Results: The two higher dosages of vitamin E were associated with a higher sperm count, greater progressive motility, and improved sperm morphology (p<0.05). These benefits were also evident after sperm freezing (p<0.05). Although chromatin abnormalities increased following vitrification, the treatment groups showed better outcomes (p<0.05). The tubular diameter, epithelium height, and luminal diameters remained unchanged with age. The tissue antioxidant capacity was greater in the groups receiving the high doses of vitamin E. Additionally, significant increases in inhibitor of DNA binding protein-4 (Id4) and GDNF family receptor alpha-1 (Gfra1) expression were observed in the higher vitamin E dosage groups, and promyelocytic leukemia zinc finger protein (Plzf) expression was notably present in the 400 mg/kg treatment group compared to the control group (p<0.05). Conclusion: Antioxidant supplementation might enhance reproductive outcomes in aging males. The observed effects included improved sperm cryo-resistance, which is advantageous for future applications such as sperm freezing or fertility preservation.

Reinke's Edema and Risk Factors, A Case-Control Study.

Reinke's edema (RE) is a benign pathological non-inflammatory disorder of the vocal folds with a wide range of clinical manifestations. We aim to investigate the relationship between Reinke's edema and some common inhalant abuse. In this case-control study, subjective consisted of 23 patients with RE (the cases), and 50 patients with sinusitis (control) who underwent surgery in the Department of Otolaryngology, between 2015 and 2020. Demographic characteristics, history of some related disease, methods, and the duration of cigarette, and opium consumption were collected through the patients' files. The chi-square (χ²) test was run to analyze the differences in the categorical and, and the Independent Sample T-test was used to compare two sample means from unrelated groups. A significant level (p-value) was considered less than 0.05. The mean age was 54 ± 12 years, and 42 ± 11 years, respectively for Reinke's edema and sinusitis. More women had been recorded in the RE group, compared to men. Allergy, unprincipled use of voice and talkativeness, history of laryngeal surgery, and type of disease were correlated to RE (p < 0.05). Also, cigarette smoking was significantly correlated with Reinke's edema. The average number of cigarettes per day, the duration of smoking, and opium consumption were more frequent in RE (P < 0.05). 90% of the RE and 4% of sinusitis patients were opium consumers. There was a statistically significant difference in the methods of substance use in the two groups of cases and control (p < 0.0001). Among the different methods, the poker and stone method was the most common (69.6%), and the opium smoking pipe was the second most common method. This study also confirmed the hazardous effects of smoking and inhaling opiates in the formation of lesions of the pharynx and larynx. In particular, longer use of these substances will be associated with more serious side effects. Therefore, it seems that people who are addicted to opiates should undergo periodic visits and counseling to reduce and stop their use.

Bone tissue regeneration by 58S bioactive glass scaffolds containing exosome: an in vivo study.

Exosomes, the naturally secreted nanocarriers of cells, have recently been demonstrated to have therapeutic benefits in a variety of disease models where parent cells are not present. However, the use of exosomes in bone defect regeneration has been unusual, and little is documented about the underlying processes. In recent study we produced and characterized exosomes derived human endometrial mesenchymal stem stromal cells and 58S bioactive glass scaffolds; in following, in this research exosome loaded scaffolds synthetized and release of exosome, porosity and bioactivity of them were assessed. More over the effect of scaffolds on repair of critical-size bone defects in rat's calvaria was evaluated by histological examination and micro computed tomography (µ CT). The findings confirmed that constructed porous scaffolds consistently release exosomes; additionally, in vivo findings including Hematoxilin & Eosin staining, Immunohistochemistry, Masson's trichrome, histomorphometric analysis, and µ CT clarified that our implant has osteogenic properties. We discovered that Exo-treated scaffolds might promote osteogenesis especially compared to pure scaffolds, indicating that produced scaffolds containing exosomes could be a potential replacement in bone tissue engineering.

Therapeutic potential of testosterone on sperm parameters and chromatin status in fresh and thawed normo and asthenozoospermic samples / Potencial terapéutico de la testosterona sobre los parámetros espermáticos y el estado de la cromatina en muestras frescas y descongeladas de normo y astenozoospermia

Background: Hormonal changes alter the physiological level of ROS and cause oxidative stress in the cell. As estimated, hormonal deficiencies, environmental and ideological factors make up about 25% of male infertility. Pathogenic reactive oxygen species (ROS) is a chief cause of unexplained infertility. Limited studies exist on the effects of testosterone on human sperm culture. Therefore, in the current study, the effect of different doses of testosterone on sperm parameters and chromatin quality was investigated. Materials and methods: Semen samples from 15 normospermic and 15 asthenospermic patients were prepared by swim up method, and then were divided into four groups by exposing to different concentrations of testosterone (1, 10, and 100nM) for 45min. Samples without any intervention were considered as control group. All samples were washed twice. Sperm parameters and chromatin protamination were assessed in each group and the remains were frozen. After two weeks, all tests were repeated for sperm thawed. Also, the MSOM technique was used to determine the sperm morphology of class 1. Results: Although sperm parameters were not show any significant differences in normospermic and asthenospermic samples exposed to different concentrations of testosterone before and after freezing, chromatin protamination was significantly decreased in the normospermic samples exposed to 10nM of testosterone before freezing (p<0.006), as well as 1 and 10nM of testosterone after freezing compared to control samples (p=0.001 and p=0.0009, respectively). (AU)

Antecedentes: Los cambios hormonales alteran el nivel fisiológico de las especies reactivas de oxígeno (reactive oxygen species [ROS]) patógenas y provocan estrés oxidativo en la célula. Según estimaciones, las deficiencias hormonales, los factores ambientales y los ideológicos constituyen alrededor del 25% de la infertilidad masculina. Las ROS son una causa principal de infertilidad inexplicable. Existen estudios limitados sobre los efectos de la testosterona en el cultivo de esperma humano. Por lo tanto, en el estudio actual se ha investigado el efecto de diferentes dosis de testosterona sobre los parámetros del esperma y la calidad de la cromatina. Materiales y métodos: Se prepararon muestras de semen de 15 pacientes normospérmicos y 15 astenospérmicos mediante el método swim up, y luego se dividieron en cuatro grupos exponiéndolos a diferentes concentraciones de testosterona (1, 10 y 100nM) durante 45min. Las muestras sin ninguna intervención se consideraron como grupo control. Todas las muestras se lavaron dos veces. En cada grupo se evaluaron los parámetros espermáticos y la protaminación de la cromatina, y los restos se congelaron. Dos semanas después se repitieron todas las pruebas de esperma descongelado. Asimismo, se utilizó la técnica MSOM para determinar la morfología espermática de clase 1. Resultados: Aunque los parámetros espermáticos no mostraron diferencias significativas en las muestras normospérmicas y astenospérmicas expuestas a diferentes concentraciones de testosterona antes y después de la congelación, la protaminación de la cromatina disminuyó significativamente en las muestras normospérmicas expuestas a 10nM de testosterona antes de la congelación (p<0,006), así como a 1 y 10nM de testosterona después de la congelación, en comparación con las muestras de control (p=0,001 y p=0,0009, respectivamente). (AU)

Therapeutic potential of testosterone on sperm parameters and chromatin status in fresh and thawed normo and asthenozoospermic samples.

BACKGROUND: Hormonal changes alter the physiological level of ROS and cause oxidative stress in the cell. As estimated, hormonal deficiencies, environmental and ideological factors make up about 25% of male infertility. Pathogenic reactive oxygen species (ROS) is a chief cause of unexplained infertility. Limited studies exist on the effects of testosterone on human sperm culture. Therefore, in the current study, the effect of different doses of testosterone on sperm parameters and chromatin quality was investigated. MATERIALS AND METHODS: Semen samples from 15 normospermic and 15 asthenospermic patients were prepared by swim up method, and then were divided into four groups by exposing to different concentrations of testosterone (1, 10, and 100nM) for 45min. Samples without any intervention were considered as control group. All samples were washed twice. Sperm parameters and chromatin protamination were assessed in each group and the remains were frozen. After two weeks, all tests were repeated for sperm thawed. Also, the MSOM technique was used to determine the sperm morphology of class 1. RESULTS: Although sperm parameters were not show any significant differences in normospermic and asthenospermic samples exposed to different concentrations of testosterone before and after freezing, chromatin protamination was significantly decreased in the normospermic samples exposed to 10nM of testosterone before freezing (p<0.006), as well as 1 and 10nM of testosterone after freezing compared to control samples (p=0.001 and p=0.0009, respectively). Similarly, chromatin protamination in the asthenospermic samples was significantly decreased at concentration of 1nM of testosterone before and after freezing (p=0.0014 and p=0.0004, respectively), and at concentration of 10nM of testosterone before and after freezing (p=0.0009, p=0.0007) compared to control samples. CONCLUSION: Using a low dose of testosterone in the sperm culture medium, has positive effects on chromatin quality.

Ischemia and vasculogenesis after transplantation of frozen and vitrified human ovarian tissue onto Chick Embryo Chorioallantoic Membrane (CAM).

OBJECTIVES: The first days of post-ovarian transplantation are critical periods, as the ischemic injury can diminish the success rate. In this study, the first day's events of ovarian transplantation in two dimensions of structure and ultrastructure following slow freezing and vitrification were assessed. STUDY DESIGN: Ovarian tissues (OTs) from 10 cancerous patients were frozen in two methods of slow freezing and vitrification. Tissues were transplanted onto the CAM and then retrieved at 5 and 10 days of culture. Nine groups were assigned as follows; I-III; fresh, 5 and 10 days culture, IV-VI; vitrification, 5 and 10 days culture, and VII-IX; slow freezing, 5 and 10 days culture. Structural and ultra-structural studies were done to assess the tissue viability and integrity following CAM transplantation. Image J software was used to measure the amounts of fibrosis and necrosis. RESULTS: The first sign of successful transplantation was found on day 3 post-transplantation. Vitrified tissues showed higher viability and transplantation rate compared to the slow frozen group (65% vs 57.5%) (p = 0.7). Tissue fibrosis and areas didn't increase significantly after cryopreservation using two methods (p > 0.05). The areas of fibrosis and necrosis and avian vessels increased significantly after 5 and 10 days of culture (p < 0.05). Large ultra-structural follicular deformities were noticed after 10 days of CAM transplantation. Better stromal ultrastructure features can be found after vitrified tissue culture. Also, the CAM transplantation technique had negative effects on the integrity of follicles, independent of the freezing procedure. CONCLUSION: Evaluation of early events of the ovarian post-transplantation is of amount importance, since the hypoxia during this period may accelerate follicular pool depletion, before the tissue stability. Vitrification can be considered a reliable alternative for slow freezing. CAM transplantation is a good technique for confirmation of tissue viability after warming but damaged the follicle ultrastructure in a short period.

Prevalence of early and late menopause and its determinants in Rafsanjan cohort study.

Our investigation aimed to evaluate the prevalence of early and late menopause and its determinants in adult women of Rafsanjan cohort study. We used data obtained from the Rafsanjan Cohort Study, as a part of the prospective epidemiological research studies in Iran. In this cross-sectional research, 2002 postmenopausal women were included in the present study. Menopause age were divided into three groups (≤ 41 years, 42-54 years, and ≥ 55 years) based on the 10th and 90th percentile. The association between age at menopause with demographic and reproductive characteristics and some clinical risk factors of women was evaluated by logistic regressions. The mean age at menopause among the study participants was 48.63 ± 5.37 years. In this study, 11.49% and 11.39% of the women experienced early and late menopause respectively. After adjusting for all potential confounders, the results showed that taller and smoker women had higher odds of early menopause (OR 1.03, 95% CI 1.00-1.06) and OR 1.85, 95% CI 1.01-3.41) respectively) and women with history of using hormonal contraceptive more than median had lower odds of early menopause (OR 0.61, 95% CI 0.41-0.91). Also older women (OR 8.65, 95% CI 5.31-14.08) and women with a history of diabetes (OR 2.42, 95% CI 1.63-3.60), hypertension (OR 2.06, 95% CI 1.42-2.97), thyroid disease (OR 1.85, 95% CI 1.07-3.20) and depression (OR 2.00, 95% CI 1.35-2.97) had higher odds of late menopause. The results showed that the year of birth, height, smoking, history of diabetes, hypertension, thyroid disease and depression and using hormonal contraceptive were significantly associated with the menopausal age. Since age at menopause can affect subsequent health in women, understanding the determinants of menopausal age is important and should be pursued.

Ultrastructure of human ovarian tissues and risk of cancer cells re-implantation after transplantation to chick embryo chorioallantois membrane (CAM) following vitrification or slow freezing.

Ovarian follicle depletion and premature ovarian failure are significant challenges in cancer patients subjected to radio- or chemotherapy. Ovarian tissue (OT) cryopreservation would be an option when other fertility preservation methods are not accessible. This study aimed to analyze the structure and ultrastructure of human OTs transplanted onto chick embryo chorioallantois membrane (CAM) after cryopreservation by vitrification or slow freezing. OTs from 10 cancer patients underwent cryopreservation. CAM transplantation was done on fresh and cryopreserved OTs, to assign samples to nine study groups as follows: 1) FI-FIII = fresh, 5- and 10-days post-CAM transplantation groups; 2) VI-VIII = vitrified, 5- and 10-days post-transplantation vitrified groups; 3) SFI-SFIII: slow frozen, 5- and 10-days post-transplantation slow freezing groups. Proliferation ability, folliculogenesis, and structural and ultrastructure were analyzed. The density of primordial follicles did not change after both freezing methods, but reduced after 5 (P ≥ 0.05) and 10 days (P ≤ 0.05) post-CAM transplantation. The follicular grade significantly decreased in all transplanted tissues (P ≤ 0.0). The proliferation marker increased after cryopreservation, but reduced after transplantation (P ≤ 0.05). TEM evaluation showed better follicular ultrastructure in the fresh group, after transplantation. Stromal ultrastructure appeared more preserved after vitrification compared with slow freezing. There was no sign of malignant cell contamination after transplantation. Some follicular TEM abnormalities were found in both methods of freezing, with a better transplantation rate after vitrification. Also, enhanced follicular activation resulted in faster follicular depletion in this method. The information regarding post grafting events would improve our knowledge for longer OTs' lifespans.

The effect of pentoxifylline and calcium ionophore treatment on sperm cell biology in oligoasthenoteratozoospermia samples.

The objective of this study was to assess the effects of pentoxifylline (PTX) and Ca2+ ionophore (CI) A12387 treatment on some biological characteristics of sperm cells in oligoasthenoteratozoospermia (OAT) patients. After processing, each sample was divided into four groups: 1, control; 2, exposed to 3.6 mM PTX; 3, exposed to 5 µm calcium ionophore (CI); and 4, exposed to both PTX and CI; 30 min at 37°C. Sperm motility was measured before and after preparation. Acrosome reaction (AR), status of sperm vacuoles, mitochondrial membrane potential (MMP) and DNA fragmentation were assessed using PSA-FITC staining, motile sperm organelle morphology examination (MSOME), JC-1 staining and sperm chromatin dispersion (CSD) test, respectively. Treatment with PTX and CI led to increased and decreased sperm motility, respectively (P < 0.05). Furthermore, vacuole status and rates of sperm DNA fragmentation were not significantly different among groups (P > 0.05). Moreover, the data showed that the rates of AR and disrupted MMP were significantly different between groups (P < 0.05). In conclusion, in vitro application of PTX not only did not have any adverse effects on sperm cell biology characteristics, but also can rectify the harmful effect of CI.

High efficiency of homemade culture medium supplemented with GDF9-ß in human oocytes for rescue in vitro maturation.

OBJECTIVE: Optimizing culture media for the incubation of immature oocytes is a vital strategy to increase the oocyte maturation rate during in vitro maturation (IVM) programs. This study evaluated the IVM and fertilization rates of human germinal vesicle (GV) and metaphase I (MI) oocytes using two different maturation media (commercial and homemade) with or without growth differentiation factor 9-ß (GDF9-ß). supplementation. METHODS: Immature oocytes from intracytoplasmic sperm injection (ICSI) cycles were collected and assigned to one of two IVM culture media (commercial or homemade; cleavage-stage base). After maturation, MII oocytes were examined under an inverted microscope for the presence of the polar body, zona pellucida (ZP) birefringence, and meiotic spindle (MS) visualization after maturation in four conditions (commercial or homemade medium, with or without GDF9-ß. ICSI was done for matured oocytes, and fertilization was confirmed by the visualization of two distinct pronuclei and two polar bodies. RESULTS: No significant differences were found between the two culture media in terms of the time and rate of oocyte maturation or the rate of fertilization (p>0.05). Growth factor supplementation increased the 24-hour maturation rate for both GV and MI oocytes only in homemade medium. The maturation rate after 24 hours was higher for MI oocytes (p<0.05). Similar results were observed for MS visualization and ZP structure in both types of media (p>0.05). CONCLUSION: Higher rates of oocyte maturation and fertilization were observed after application of homemade medium supplemented with GDF9-ß. Therefore, this combination may be recommended as an alternative for clinical IVM programs.

Comparing the IVM laboratory outcomes between stimulated IVF with unstimulated natural cycles.

Recently, more attention has been raised towards fertility preservation in women with cancer. One option is in vitro maturation (IVM) of the immature oocytes as there is not enough time for induction of an ovarian stimulation protocol. The aim was to compare the IVM laboratory outcomes between stimulated and unstimulated (natural) in vitro fertilization (IVF) cycles. In total, 234 immature oocytes collected from 15 cancer patients who underwent an IVM programme (natural IVM) and 23 IVF cycles with a controlled ovarian hyperstimulation protocol (stimulated IVM) were analyzed. The oocyte morphology, zona pellucida (ZP), and meiotic spindle presence were measured using PolScope technology. Also, the rates of oocyte maturation and fertilization were assessed in both groups. The IVM rate was higher in the stimulated cycle (P < 0.05), but the fertilization rate was insignificant in comparison with unstimulated cycles. There were no significant differences in the spindle visualization and ZP birefringence scoring between the groups (P > 0.05). The oocyte normal morphology was better in the stimulated cycle compared with the natural cycle (P < 0.05). In conclusion, IVM can be recommended for cancer patients as an alternative treatment when there is insufficient time for conventional IVF before chemotherapy initiation.

Investigation of the Effect of Prunus Amygdalus Amara on the Expression of some Genes of Apoptosis and Immortality in Breast Cancer Cells (MCF- 7).

BACKGROUND: Anti-cancer effects of almond nuts or oil have been approved, but there are a few pieces of research that have evaluated, in detail, almond and other seeds' effects on cancer. Therefore, in the present project, the aim was to explore the regulatory effect of the bitter almond extract (Prunus amygdalus Batsch) on the apoptotic and anti-cancer potency of MCF-7 cells. OBJECTIVE: In the current experimental research, the almond effect on MCF7 cells was evaluated by investigating the expression and the balance between Bcl-2, Bax genes to unmark the potential molecular mechanism. METHODS: For 24 and 48h, the MCF7 cells were treated with the bitter almond extract (187.5-3000 µg/mL). MTT assay was used to assess the viability, and Real-time-PCR was applied to determine the expression of Bax and Bcl-2, facing ß-actin. RESULTS: Our results revealed a significant difference between different extract concentrations on the viability of MCF7 cell lines in 24 and 48 h; cell viability decreased time-dependently (P < 0.05). After 24 and 48h of extract facing MCF7 cells, the evaluated IC50 value was 3000 and 1500 µg/mL, respectively. Based on Real-Time-PCR analysis, after 24 and 48 h, the mRNA levels of BCL-2 decreased by the extract, whereas Bax was in the MCF-7 cell line. CONCLUSION: From the results, it can be concluded that bitter almond extract has anti-cancer properties that may influence the apoptotic pathways by regulating relative gene expression.

Resilience as the predictor of quality of life in the infertile couples as the most neglected and silent minorities.

BACKGROUND: It has been demonstrated that infertility can affect quality of life (QoL) in infertile couples. Resilience is considered a protective factor against the distress caused by infertility and the quality of life status. There is a new definition for Fertility Quality of Life that evaluates particularly the impact of infertility on various aspects of life. MATERIAL AND METHODS: In this couple-based study, the main objective was investigating the quality of life based on the gender and resilience of infertile couples. Measurement tools were three questionnaires including a demographic one, a quality of life of infertile couples questionnaire and the Connor-Davidson Resilience Scale. Data analysis was done through paired t-test and linear multiple regressions test. RESULTS: Overall the difference of mean score for QoL is statistically significant (P > 0.001) between men and women (69.48% vs 58.87%), which means that QoL status was positive in men and neutral in women. In addition, the mean score of male resilience was more than female resilience (P = 0.009). The results showed there was a significant and positive correlation between the QoL status and resilience score (P = 0.008, r = 0.13) (P < 0.1), and resilience (ß = 0.04 and P = 0.04) had a significant protective effect on the quality of life. CONCLUSION: Low resilience status in infertile couples is better to be considered as a risk factor compromising the quality of life and infertility consolers should keep in mind this issue and provide a comprehensive and holistic approach for a better outcome of infertility treatment. ABBREVIATIONS: QoLICQ: Quality of Life in Infertile Couple Questionnaire; CD-RISC: Connor-Davidson Resilience Scale; IVF: in vitro fertilisation; ART: assisted reproductive technique; PTSD: posttraumatic stress disorder; IUI: intrauterine insemination; ICSI: intracytoplasmic sperm injection.

Proliferating cell nuclear antigen presentation, as a marker of folliculogenesis, in the transplanted ovarian tissue.

AIM: One of the most important ways to understand the ovarian biology is studding the initiation of primordial follicle development and subsequent folliculogenesis control. In this study, proliferating cell nuclear antigen (PCNA) presentation was used as a marker of follicular development in the thawed ovarian tissue (OT) following transplantation onto chick embryo chorioallantoic membrane (CAM) using two methods of freezing of slow freezing and vitrification. METHODS: Samples of OT from 10 patients were subjected to slow freezing and vitrification. After warming, CAM transplantation was done and PCNA proliferation index (PI; percent of PCNA-positive granulosa cells) was calculated for each follicle stage. Image J software was used to determine the mean staining intensity. RESULTS: PCNA was positive for granulosa cells and oocytes nuclei, but negative for ooplasm. There were no remarkable PCNA staining in the granulosa cells of primordial follicles, but increased significantly as follicle progression (p < 0.05). Proliferation rate was also insignificantly higher in the vitrified than slow freezing group, before and after transplantation (p < 0.05). Lower PCNA presentation index was observed after CAM transplantation (p < 0.05). The earliest stage of follicular recruitment took place in the transitional follicles, before squamous cells transform to cuboidal cells. CONCLUSION: PCNA showed that follicles had proliferation power after cryopreservation. Higher presentation after vitrification may indicate accelerated folliculogenesis in the thawed OT.

Ovarian Tissue Reservation and Risk of Reimplantation in a Young Girl with Acute Lymphocytic Leukemia after 6-Month Chemotherapy: A Case Report.

In the fertility preservation programs, ovarian cryopreservation is usually offered when the risk of premature ovarian failure is high (>30-50%) while the risk of ovarian metastasis is low. According to the guidelines, it must be done before the patient receives chemotherapy. A 22-year-old girl with acute lymphocytic leukemia was a candidate for ovarian cryopreservation after 6 months of chemotherapy. Despite chemotherapy, the anti-Mullerian hormone survey was within normal range. Ovarian tissue cryopreservation was done. In the histology survey, follicular density was 7.48. This case shows that only having a history of chemotherapy does not exclude the patient from the fertility preservation program. Regarding referring the patients for fertility preservation, cumulative factors such as a history of gonadotoxic treatment, age, and treatment protocol should be considered. In addition, the case was negative for assessing of CD45 marker. New data may challenge previous strict criteria, and extend the indications of this effective method in preserving fertility among cancer patients.

The effect of testosterone and antioxidants nanoliposomes on gene expressions and sperm parameters in asthenospermic individuals.

BACKGROUND: Antioxidants that used for the infertility treatment cannot have their complete effectiveness, because of their instability in the culture medium. SIGNIFICANCE: One of the most advances, in the drug delivery systems, is nanoliposomes-loaded, as biodegradable and bioavailable carriers. Hormonal and antioxidant agents encapsulating inside the nanoliposomes were used, to increase the effectiveness of antioxidants in the sperm culture medium. MATERIALS: Semen sample from 15 asthenospermia were divided into 10 equal parts. After preparation, the sperms were incubated with free form of drugs and nanocarriers contained resveratrol, catalase, resveratrol-catalase and testosterone for 45 min. All sperm parameters, sperm DNA and gene expressions were evaluated before and after freezing. RESULTS: Before freezing, all nanocarriers and free testosterone showed higher sperm motility compared to free drugs (p=.000). Free Testosterone and free resveratrol-catalase had higher DNA damage compared to nanocarriers (p=.000). Before freezing, the blank nanoliposome and testosterone nanoliposomes had the lowest HSP70 gene expression respectively (p = 0.005) (p = 0.001). After freezing, a significant reduction in sperm motility was observed in the free resveratrol-catalase group (p=.003). Also, a significant increase in sperm viability was observed in the free testosterone and nanoliposomes of blank and testosterone (p > 0.05). The least DNA fragmentation was related to catalase nanoliposomes (p=.000). All nanoliposomes, especially catalase, had the highest percentage of class I morphology compared to the control group (p=.000). CONCLUSIONS: Nanoliposomes could improve the sperm parameters and DNA integrity before and after freezing, by increasing the effectiveness of antioxidants. So, it can be recommended in the ART lab.

Effective dosage of growth differentiation factor-9ß in folliculogenesis and angiogenesis in the sheep ovarian tissues grafted onto chick embryo chorioallantoic membrane.

AIM: Scientists have tried to culture and transplant the ovarian tissues (OT), to preserve fertility in cancer patients. However, one of the main limitations to the applicability of this technique is the folliculogenesis disruption after transplantation. Due to the roles exerted by growth differentiation factor-9ß (GDF9ß), we decided to determine the most effective dose of GDF9ß on promotion of folliculogenesis and angiogenesis in sheep OT grafted onto the chick chorioallantoic membrane (CAM). METHODS: Fresh sheep OT were grafted onto the CAM for 5 days, and divided into four groups based on the addition of increasing doses of GDF9ß (0, 150, 200 and 250 ng/mL). Following culture, histological (hematoxylin and eosin [H&E] staining) and immunohistological studies (Ki-67) were done. Fibrotic and necrotic regions were measured using MICROVISIBLE software. For comparing the follicle development rates between the groups as well as differences in the Ki-67-positive follicles, analysis of variance was applied. RESULTS: In both 200 and 250 ng/mL GDF9ß groups, significantly higher rates of intermediary and primary follicles were observed, also the numbers of good quality follicles increased in the aforementioned groups and the rates of fibrotic and necrotic areas decreased. Moreover, in the 200 and 250 ng/mL GDF9ß groups, the number of capillaries and the proliferative activity increased. The lower dose of GDF9ß (150 ng/mL) neither activated the primordial follicles nor lead to an increase in the number of growing follicles. CONCLUSION: Addition of high dosages of GDF9ß to the OT, grafted onto the CAM resulted in higher folliculogenesis and better transplantation features due to improvement in angiogenesis.

Vitrification has detrimental effects on maturation, viability, and subcellular quality of oocytes post IVM in cancerous women: An experimental study.

BACKGROUND: In vitro maturation (IVM) of immature oocytes retrieved from ovarian tissue has been considered as a valuable approach for fertility preservation in cancerous patients. OBJECTIVE: To evaluate the efficacy of vitrification on oocyte maturation, survival rates, as well as the subcellular oocyte quality post IVM. MATERIALS AND METHODS: The ovarian cortexes from 19 women with cervix and uterine malignancy aged 21-39 yr were collected. Cumulus-oocyte complexes were aspirated from all visible antral follicles. 102 immature oocytes were collected, and 43 oocytes were detected appropriately for IVM (control group). Also, 59 immature oocytes were vitrified, then matured in vitro (IVM) in two groups: with Growth/differentiation factor 9 (GDF9) (group 1) and without GDF9 (group 2) supplementation. Rates of oocytes viability, maturation, and survival along with meiotic spindle visualization and zona pellucida birefringence were assessed with Polyscope. RESULTS: The rate of maturation was significantly higher in controls (55.8%) compared to the other groups. Maturation rate was 23.3% in oocytes cultured in IVM medium enriched with GDF9, and 27.6% in those cultured in IVM medium lacking GDF9 (p = 0.86). Also, the meiotic spindle was present in 74.4% of control oocytes which was significantly higher than the other groups. The proportion of high zona pellucida birefringence was higher in the controls when compared with group 1 (51.2% vs. 23.3%, respectively, p = 0.04). CONCLUSION: Vitrification had a detrimental effect on oocyte maturation, viability as well as the subcellular quality of the oocytes after IVM in cancerous women.

Predictors of Quality of Life in Infertile Couples.

OBJECTIVES: This study was conducted with the aim of examines the quality of life of infertile couples and their relationship with the practical resilience of infertile couples referring to Yazd's centers of infertility. METHODS: This research is a descriptive-correlational study. The research population consisted of all infertile couples who referred to Infertility Centers in Yazd, Iran in the winter of 2016. Sampling was conducted in a non-random and accessible manner. The instrument used in the research included a) demographic information questionnaire, b) Conner and Davidson's Resilience Scale, and c) quality of life infertile couples questionnaire. Data were analyzed by SPSS software version 17 at a significant level of P < 0.05. To describe the data, descriptive statistics methods were used and the inferential statistics (Pearson correlation coefficient, regression, independent t test, and variance analysis) were used to test the research hypotheses. RESULTS: People (202 couples) participated in this research. Three variables of resilience (ß = 0.04, P = 0.04), gender (ß = -0.22, P < 0.001), and education level (ß = 0.21, P < 0.001) had a prediction coefficient and there was a significant relationship with quality of life. CONCLUSIONS: This study showed that resilience, gender, and education predict the quality of life of infertile couples. In the infertile couples counseling program, resilience should be considered as a coping factor.

GDF9-ß promotes folliculogenesis in sheep ovarian transplantation onto the chick embryo chorioallantoic membrane (CAM) in cryopreservation programs.

PURPOSE: Ovarian tissue (OT) cryopreservation is a treatment option for fertility preservation among young cancer patients. However, the procedure may involve a reduction in the GDF9-ß expression and a delay in follicular growth after thawing and transplantation. The aim of this study was to evaluate whether supplementation of GDF9-ß can compensate the reduction of this factor during the cryopresevation process and promote folliculogenesis after transplantation of thawed sheep ovarian tissue. METHODS: Sheep OT was cryopreserved using two methods of vitrification and slow freezing. Fresh and thawed OTs were then transplanted onto chick embryo chorioallantoic membrane (CAM) and then divided into two groups based on the addition of GDF9-ß to the grafted tissue. After 5 days of culture, both histological and immunohistological (Ki-67) assessments were performed to evaluate follicular structure, development, and proliferation. The fibrotic and necrotic areas were measured using MICROVISIBLE software. RESULTS: Folliculogenesis took place in all culture groups, but was significantly improved only in the +GDF9-ß cultured group. Also, better follicular structure was preserved in the aforementioned group (p < 0.05). When GDF9-ß was supplemented to the culture medium, more neovascularization (p < 0.05) and better transplantation (p > 0.05) was observed. Furthermore, the areas of fibrosis and necrosis were lower in this group rather than the controls. Follicular proliferative activity was significantly higher only in the slow freezing +GDF9-ß cultured group. CONCLUSIONS: GDF9-ß, as a stimulatory factor, not only promoted the folliculogenesis in the fresh ovarian transplant, but also compensated for its reduction during the cryopreservation process.