Association between follicular fluid bacteria with inflammatory markers of the complete blood count and the outcomes of assisted reproductive technology in women with endometriosis: A case-control study.

Background and Aims: Endometriosis is a common reason for infertility and poor outcomes of assisted reproductive technology (ART). Inflammation is involved in the pathogenesis of this disease. The presence of microorganisms in women with endometriosis may increase levels of inflammatory markers. The purpose of this study is to determine the relationship between the presence of bacteria in the follicular fluid with the inflammatory markers of the complete blood count (CBC) and the outcomes of in vitro fertilization (IVF) in women with endometriosis. Methods: This case-control study was conducted on 74 patients undergoing IVF, referred to Al-Zahra Hospital in Rasht (Iran) in 2021. The patients were divided into two case groups including 37 women with endometrioma and the control group, including 37 infertile women with a male factor and normal ultrasound. In total, 74 follicular fluids were collected from the case and control groups and were cultured in the laboratory. The relationship between culture results with IVF outcomes and the levels of CBC inflammatory markers including the number of white blood cells (WBCs), lymphocytes, neutrophils, neutrophil-to-lymphocyte ratio (NLR), lymphocyte-to-monocyte ratio (LMR), and platelet-to-lymphocyte ratio (PLR), erythrocyte sedimentation rate (ESR), and c-reactive protein (CRP) was analyzed. Results: There was no significant statistical difference between the frequency of bacteria present in the follicular fluid (p = 0.861), the mean rate of fertilization (p = 0.363), the frequency of CRP (p = 0.999), and the mean WBCs, lymphocytes, neutrophils, NLR, LMR, and PLR in the two groups. There was a significant statistical difference between the mean number of oocytes of metaphase II (p = 0.034) and the mean ESR (p = 0.018) in the two groups. Conclusions: It seems necessary to evaluate follicular fluid as a biological substance that is considered an optimal factor for predicting oocyte quality, fertilization rate, embryo quality, and the success rate of ART.

Effects of cryopreservation in the presence of Natural Deep Eutectic Solvents (NADESs) on sperm parameters.

Natural Deep Eutectic Solvents (NADESs) are being considered as a potential alternative to traditional cryoprotective agents (CPAs) in sperm freezing. The study aimed to assess the effects of NADESs as a CPA on human sperm parameters. A total of 32 normozoospermic semen samples were collected from the Alzahra infertility treatment center (Iran) between July 2021 and September 2022. The samples were categorized into eight different groups: 1) a control (nonfrozen), and groups frozen with 2) SpermFreeze Solution, 3) ChX (Choline chloride and Xylitol), 4) ChS (Choline chloride and D-sorbitol), 5) ChG (Choline chloride and Glucose), 6) ChU (Choline chloride and Urea), 7) EtP (Ethylene glycol and l-proline), and 8) GlyP (Glycerol and l-proline). The study also analyzed the quality of sperm parameters, such as chromatin condensation and integrity, acrosome integrity, and survival, along with the expression of some genes that affect sperm fertility (TRPV1, TRPV4, SPACA3, and OGG1). The study found there were notable variations in sperm parameters (such as viability, chromatin condensation and integrity, and acrosome integrity) among frozen groups with some NADESs compared to the SpermFreeze Solution and control groups (P < 0.05). Analysis of gene expression demonstrated that the levels of TRPV1, TRPV4, SPACA3, and OGG1 genes were superior in the GlyP group compared to the other groups (P < 0.05). Additionally, the ChS and ChU groups exhibited preserved expression of these genes compared with the SpermFreeze Solution group. The use of NADESs led to the discovery of a more appropriate CPA that has low toxicity and is highly effective in maintaining the fertility potential of sperm.

The Association of Soluble VEGFR-1 Serum Level and Genetic (rs7993418) Polymorphism with In Vitro Fertilization and Embryo Transfer Outcome in the Population of Northern Iran.

Background: Vascular endothelial growth factor receptors (VEGFRS) play an important role in embryo implantation. The aim of the present study was to examine the association of VEGFR1 circulating level and gene polymorphism with in vitro fertilization and embryo transfer (IVF-ET) outcome. Methods: In this case-control study, 120 women who had unsuccessful IVF (IVF-) history and 120 women who had successful IVF outcome (IVF+) as controls were included. Genomic DNA was extracted from blood samples. Genotyping was performed using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). The serum levels of soluble VEGFR1 (sVEGFR1) were measured by ELISA. ANOVA test was used for statistical analysis. Results: The frequency of T and C alleles in IVF+ individuals were 87.5%, 12.5% and among IVF- were 75.5%, 24.5%, respectively (p=0.0006). The minor allele (C) was associated with an increased risk of IVF failure based on results from co-dominant (OR=3.86, 95%CI 1.19-12.47), dominant (OR=2.32, 95%CI 1.31-4.10), recessive (OR=3.22, 95%CI 1.00-10.29), and allele models (OR=2.28, 95%CI 1.40-3.69). We also showed that there is a significant decrease in serum sVEGFR1 levels in IVF as compared to IVF+ (p=0.006) groups. Moreover, TT genotype is significantly associated with increased serum sVEGFR1 concentration in IVF group (TT, CT, and CC serum levels were 106.55±11.04, 94.33±10.75, and 83.33±9.13 ng/ml, and in IVF+ group were 156.11±18.08, 120.66±16.51, and 84.66±20.31 ng/ml, respectively). Conclusion: The results of this study indicate that VEGFR1 polymorphism and sVEGFR1 circulating levels are associated with IVF-ET outcome. Moreover, CC genotype is associated with decreased sVEGFR-1 serum concentration and IVF-ET failure.

CRISPR/Cas9-mediated knockout of HO-1 decreased the proliferation and migration of T47D cells and increased cisplatin-induced apoptosis: an in vitro study.

Breast cancer is the most common type of neoplasm and the second cause of cancer-related death in women. Despite the development of novel therapeutic strategies and improved the clinical outcomes, the mortality rate for breast cancer is still high. Therefore, development of a new modality, particularly based on knocking out key genes, is under focus of investigation. Heme oxygenase-1 (HO-1) deregulation has been associated with various neoplasms-related behaviors of many types of tumor cells including breast cancer. In the current study, in order to evaluate the role of the HO-1 gene in breast cancer, we utilized the CRISPR/Cas9 technology to knock out HO-1 gene in T47D breast cancer cell line and studied its potential therapeutic effects in vitro. The cell proliferation and their sensitivity to Cisplatin were determined by CCK-8 kit. In addition, the apoptosis and the migratory potential of the cells were evaluated using Hoechst staining, and Transwell/Scratch methods, respectively. Our findings revealed that HO-1 suppression significantly reduced the proliferation ability of T47D cells (P < 0.001). Moreover, sensitivity to Cisplatin-induced toxicity increased significantly in KO-T47D cells compared to the control T47D cells. Furthermore, our findings indicated that Cisplatin-induced apoptosis increased in the KO-T47D cells. Moreover, the migratory capability of KO-T47D cells was abolished significantly (P < 0.001) as determined by Transwell migration assay. In a nutshell, our findings strongly suggest that HO-1 involved in breast cancer progression and metastasis and chemotherapy resistance. However, further comprehensive studies are required to clarify the precise role of the HO-1 gene on breast cancer cells.

An In vitro Study on the Protective Effect of Melatonin on Human Sperm Parameters Treated by Cadmium.

Background: Male infertility account for nearly 50% of infertility cases. Cadmium is regarded as a well-known toxic metal for industrial applications; high amounts of cadmium in the human body can result in chronic toxicity. Melatonin as a free radical scavenger has anti-inflammatory, and even anti-cancer and antiapoptotic functions. Aim: In this work, we evaluated the protective effect of melatonin on human sperm parameters treated by cadmium. Study Setting and Design: This was an experimental study carried out from May to December 2019. Materials and Methods: A total of 41 fresh semen samples were collected from fertile men and were divided into 4 groups: (1) control, (2) sperm +25 Nm cd, (3) sperm +25 nM cd +0.1 mM melatonin,(4) sperm +0.1 mM melatonin treated for 60 min. In all groups, semen analysis was performed for motility, viability and DNA fragmentation index (DFI). Statistical Analysis: The groups were compared using the ANOVA test. Results: The group treated with cadmium showed a significant decrease in rapid and slow motility, and survival rate compared with the control group (P < 0.05). However, the degree of DFI and sperm with non-progressive motility in the group treated with cadmium had a significant increase compared to the control (P < 0.05). The use of melatonin significantly improved sperm parameters such as motility, survival rate and decreased sperm DFI with non-progressive motility. Conclusions: The use of melatonin reduces the amount of cadmium damage in human sperm in vitro.

Using Deep Learning Algorithm: The Study of Sperm Head Vacuoles and Its Correlation with Protamine mRNA Ratio.

OBJECTIVE: It is necessary to evaluate fertility effective agents to predict assisted reproduction outcomes. This study was designed to examine sperm vacuole characteristics, and its association with sperm chromatin status and protamine-1 (PRM1) to protamine-2 (PRM2) ratio, to predict assisted pregnancy outcomes. MATERIALS AND METHODS: In this experimental study, ninety eight semen samples from infertile men were classified based on Vanderzwalmen's criteria as follows: grade I: no vacuoles; grade II: ≤2 small vacuoles; grade III: ≥1 large vacuole and grade IV: large vacuole with other abnormalities. The location, frequency and size of vacuoles were assessed using high magnification, a deep learning algorithm, and scanning electron microscopy (SEM). The chromatin integrity, condensation, viability and acrosome integrity, and protamination status were evaluated for vacuolated samples by toluidine blue (TB) staining, aniline blue, triple staining, and CMA3 staining, respectively. Also, Protamine-1 and protamine-2 genes expression was analysed by reverse transcription-quantitative polymerase chain reaction (PCR). The assisted reproduction outcomes were also followed for each cycle. RESULTS: The results show a significant correlation between the vacuole size (III and IV) and abnormal sperm chromatin condensation (P=0.03 and P=0.02, respectively), and also, protamine-deficient (P=0.04 and P=0.03, respectively). The percentage of reacting acrosomes was significantly higher in the grades III and IV spermatozoa in comparison with normal group. The vacuolated spermatozoa with grade IV showed a high protamine mRNA ratio (PRM-2 was underexpressed, P=0.01). In the IVF cycles, we observed a negative association between sperm head vacuole and fertilization rate (P=0.01). This negative association was also significantly observed in pregnancy and live birth rate in the groups with grade III and IV (P=0.04 and P=0.03, respectively). CONCLUSION: The results of our study highlight the importance sperm parameters such as sperm head vacuole characteristics, particularly those parameters with the potency of reflecting protamine-deficiency and in vitro fertilization/ intracytoplasmic sperm injection (IVF/ICSI) outcomes predicting.

Effect of astaxanthin and melatonin on cell viability and DNA damage in human breast cancer cell lines.

BACKGROUND: Astaxanthin is a xanthophyll pigment found in algae and marine animals, having strong anti-oxidative, anti-tumoral, and anti-inflammatory effects. Additionally, melatonin has shown inhibitory effects on the growth of human breast cancer cells. The aim of the present study was to evaluate the effect of astaxanthin and the combined effects of astaxanthin and melatonin on breast cancer cells and the non-tumoral breast cell line. MATERIALS AND METHODS: The human breast cancer cell lines, T47D and MDA-MB-231, and non-tumorigenic cell line MCF 10A were treated and compared to astaxanthin, melatonin, and co-administration of these two compounds. Cell viability, apoptosis induction, Bcl-2 protein expression, and DNA damage were measured by MTT assay, acridine orange/ethidium bromide (AO/EB) staining, immunocytochemistry, and comet assay. RESULTS: Astaxanthin at lower doses than melatonin reduced cell viability and Bcl2 expression, induced apoptosis and DNA damage in MDA-MB-231 and T47D. Meanwhile, the effects of astaxanthin on cell cytotoxicity, apoptosis, and DNA damage in MCF10A cells are insignificant compared to MDA-MB-231 and T47D. Moreover, the results indicated that astaxanthin in T47D cells caused more cell death compared to MDA-MB-231 cells. Astaxanthin induced cell death on breast cancer cells and without cell cytotoxicity for non-cancerous cells. CONCLUSION: Furthermore, the presence of astaxanthin increased the function of melatonin-induced cell death in breast cancer cells.

Dimethyl fumarate prevents cytotoxicity and apoptosis mediated by oxidative stress in human adipose-derived mesenchymal stem cells.

BACKGROUND: The poor survival rate and undesirable homing of transplanted stem cells are the major challenges in stem cell therapy. Addressing the challenge would improve the therapeutic efficacy of these cells. Dimethyl fumarate (DMF) is an anti-inflammatory drug that exerts its effects through the activation of the nuclear factor erythroid 2-related factor 2 (Nrf2) pathway. Therefore, its cytoprotective effects on human adipose-derived MSCs (hASCs) against various oxidative stresses have been investigated in this study. METHODS AND RESULTS: hASCs were cultured with different concentrations of DMF to evaluate the cytotoxicity of DMF on hASCs using Cell Counting Kit-8 (CCK-8). Besides, the migration ability of the cells after DMF treatment was evaluated using the Transwell method. Furthermore, the expression of HO-1 and NQO-1 was determined using RT-PCR. The cytoprotective effects of DMF on hASCs against the oxidative stress caused by H2O2 and Ultra Violet (UV) were evaluated by assessing cell proliferation and apoptosis. Our results demonstrated that under oxidative stress conditions induced by H2O2 and UV, DMF increased the survival rate and proliferation of the cells and prevented apoptosis. Moreover, the expression of HO-1 and NQO-1 was upregulated in hASCs pretreated with DMF which confirms the activation of the Nrf2 pathway. However, DMF significantly decreased migration in hADSCs (P < 0.0001). CONCLUSION: Our findings indicate that DMF enhances the proliferation capability and viability of hASCs and prevents their apoptosis in harsh stressful microenvironments. However, the applicability of DMF as a cytoprotective factor for the augmentation of hASCs requires in-depth preclinical and clinical studies.

Effects of melatonin on the toxicity and proliferation of human anaplastic thyroid cancer cell line.

BACKGROUND: Thyroid carcinoma is the most common endocrine malignancy and anaplastic thyroid carcinoma (ATC) is a rare but most aggressive cancer. Melatonin has enhanced or induced apoptosis in many different cancer cells, however, there has not been any study on the effects of melatonin in the treatment of ATC. In this study, we examined the effect of melatonin on cytotoxicity in the human ATC cell line. MATERIALS AND METHODS: Cultured ATC cells were treated at melatonin concentrations 0.6, 1, 4, 16, 28 mM for 24 h. The MTT assay was performed to examine cell viability. Cytotoxicity was assayed with the determination of lactic dehydrogenase (LDH) activity. Apoptosis was detected by acridine orange/ethidium bromide and Hoechst 33342 staining. Giemsa staining is considered for evaluating the morphological changes of ATC cells. The reproductive ability of cells to form a colony was evaluated by the clonogenic assay. RESULTS: Results showed that melatonin could significantly decrease cell viability and the lowest cell viability was observed at 28 mM, 10.26 % ± 0.858 versus control. Similar results were obtained when analyzing LDH activity. The highest LDH levels were observed at 16 and 28 mM (546.08 ± 4.66, 577.82 ± 3.14 munit/mL versus control) that confirmed the occurrence of late apoptosis. The clonogenic assay showed that cells at the high concentration of melatonin (16 and 28 mM) don't enable to form the colony that approved the occurrence of reproductive death. CONCLUSION: Our results showed a dose-dependent cytotoxic effect of melatonin on ATC cells that significantly decreased cell viability and induced cell reproductive death at the concentration greater than 1 mM and findings suggested that MLT might be useful as an adjuvant in ATC therapy.

Anti-proliferative and apoptotic effects of hull-less pumpkin extract on human papillary thyroid carcinoma cell line.

Papillary thyroid carcinoma (PTC) is one of the most common cancers of the endocrine system. Previous studies have shown that the extract of hull-less pumpkin seed (HLPS) has a significant anti-cancer effect. The aim of this study was to evaluate the effect of this plant extract on the proliferation of PTC cells. In this study, an extract of this plant was prepared by soxhlet extraction method and analyzed by Gas Chromatography-Mass Spectrometry. The cytotoxicity of PTX and plant extract was investigated using the methylthiazol tetrazolium (MTT) method. For careful investigation of morphological alteration, we used hematoxylin and eosin and Giemsa stinging. Based on MTT assay test, the IC50 value of paclitaxel (PTX) was significantly less than the hydro-alcoholic extract of HLPS at all of the incubation time. Our results of histological staining showed that HLPS and PTX induced significant morphological alteration in the PTC cultured cell that consistent with cell death. Comparing the groups treated by PTX or HLPS with control group showed significant differences. It seems that HLPS extract has an apparent effect on treatment of PTC, at least in laboratory condition, albeit for realistic decision about the effect of HLPS on PTC, more molecular investigations are necessary.

The Association of PON1 192 Q/R Polymorphism with the Risk of Idiopathic Male Infertility in Northern Iran.

[This retracts the article on p. 253 in vol. 10, PMID: 30555659.].

The Association of PON1 192 Q/R Polymorphism with the Risk of Idiopathic Male Infertility in Northern Iran.

BACKGROUND: Infertility is defined as the inability to achieve pregnancy after 12 months of regular unprotected sexual intercourse. Environmental and genetic factors are involved in male infertility. The polymorphism studies have a crucial role in disease recognition. Paraoxonase (PON) is an oxidant enzyme which is associated with inflammation, oxidative stress and lipid metabolism. The present study aimed to evaluate the relationship between PON1 192 Q/R polymorphism and the susceptibility to idiopathic male infertility. METHODS: Samples were collected from 220 patients diagnosed with male infertility and 230 controls genotyped by Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP). RESULTS: A significant difference in genotype distributions of PON1 192 Q/R polymorphism was observed between patients and controls (p=0.001). Our findings revealed that individuals with the variant QR had a significant decreased risk of idiopathic male infertility (OR=0.49, 95%CI=0.33-0.73, p=0.0004). Moreover, analyses showed that R allele may have a protective effect on susceptibility of idiopathic male infertility (OR=0.31, 95%CI=0.21-0.47, p=0.0001). CONCLUSION: The data from this study indicates that the PON1 192 Q/R polymorphism is associated with decreased risk of idiopathic male infertility. However, more studies should be considered with larger number of patients and control subjects to confirm our results.

From dentistry students' points of view: do anatomy classes we took actually boost our learning during clinic?

The aim of medical education is to teach the essence of practical skills alongside with the theoretical knowledge. Teaching anatomy, as the center of medical education, should be leading to use this knowledge as a skill during clinical period. According to the rising numbers of dentistry faculties' experiences, inappropriate education results in misguidance during clinic. Thus, this study was conducted to find about the pre-clinical and clinical dentistry students' points of view on the helpfulness of anatomy classes in achieving clinical goals. Present descriptive cross-sectional study evaluated Guilan University of Medical Sciences' pre-clinical and clinical dentistry students' opinions on the effectiveness of anatomy classes during their clinical period in 2017. The sampling method used here was census via questionnaire and scoring was according to Likert scaling system. Analyses showed that anatomy of the nervous system was the most assistive course, which helped dentistry students during their clinical period (P<0.001). The least scored course was visceral organs and that means they did not use most of their learnings from classes with this topic (P<0.001). They also stated that other important factors such as using cadavers and moulages in practical sessions, teaching clinical skills theoretically before practical sessions and performing group activities are crucial for them to recall important details of the relevant courses during clinical period. Results of this study suggests that alongside with the various topics of anatomy courses, other factors like professors' characteristics and their teaching methods are also of important factors helping the dentistry students throughout clinic.

Assessment of EGFR Gene Expression Following Vitrification of 2-cell and Blastocyst Mouse Embryos.

BACKGROUND: Exact mechanisms of fetal harm following vitrification are still unknown. This study was conducted to evaluate the cryopreservation impact on the expression of Epidermal Growth Factor Receptor (EGFR) gene in mouse 2-cell and blastocysts. METHODS: To stimulate ovulation in mice, hCG was injected, followed by collecting 2-cells and blastocysts after 44-46 and 88-89 hr, respectively. These embryos were divided into two case and control groups. The fresh case group was cryopreserved using cryotop and warmed after 4 mounts. Normal 2-cells were selected based on their morphology and their RNA was extracted. Quantitative expression of EGFR gene in both groups was investigated by applying real time-PCR. RESULTS: The statistical Real-Time (RT)-PCR analyses performed using SPSS revealed that the expression level of EGFR gene was diminished in the case group compared to the control group. CONCLUSION: The current study indicated the negative effect of cryopreservation on expression amount of EGFR gene in 2-cell and blastocyst mouse embryos.

Level of Vitamin E in Follicular Fluid and Serum and Oocyte Morphology and Embryo Quality in Patients Undergoing IVF Treatment.

Objective: This study aimed to determine the relationship between vitamin E in the follicular fluid (FF) and serum with oocyte morphology and embryo quality. Materials and methods: A cross-sectional study was conducted on serum samples, FF samples, oocytes, and embryos collected from 50 women undergoing in vitro fertilization in the Alzahra Hospital, Rasht, Iran from March to August 2014. Vitamin E level was measured using HPLC. Oocyte morphology and embryo quality were evaluated during inverted optical microscopy. Results: Totally 434 oocytes and 199 embryos were examined. Most frequently the metaphase II (MII) oocytes were observed at the 0.35-1 mg/dl level of vitamin E in FF (89.2%) and the 1-5 mg/dl level of vitamin E in serum (86.1%). The odds of having MII oocytes at the level of 0.35-1 mg/dl (OR = 2.48, 95% CI = 1.24-4.94) and 1.5-2 mg/dl (OR = 2.51, 95% CI = 1.02-6.19) of vitamin E in FF was significantly higher compared to level of 2-7.4 mg/dl. The effect of vitamin E serum level on oocyte maturation was not significant. The odds of having embryo with Z1 or Z2 quality, at the 10-15 mg/dl level of vitamin E in serum (OR = 6.45, 95% CI = 1.18-35.22), compared to the 15-20 mg/dl level, was significantly higher. The effect of vitamin E levels in FF on the embryo quality was not significant. Conclusion: At certain levels of vitamin E in the FF, oocytes with higher maturation and at certain levels of vitamin E in serum, embryo with higher quality can be achieved.

Polymorphism variant of MnSOD A16V and risk of female infertility in northern Iran.

OBJECTIVE: Infertility is a disease of the reproductive system defined by inability to conceive after having regular unprotected intercourse. Both environmental and genetic factors can be involved in female infertility. Manganese superoxide dismutase (MnSOD) is a crucial mitochondrial antioxidant enzyme that has a key role in cellular defense against agents that induce oxidative stress. The present study was aimed to evaluate the MnSOD A16V gene polymorphism in female infertility in northern Iran. MATERIALS AND METHODS: Samples were obtained from 150 patients diagnosed with female infertility and 150 controls and genotyped by the polymerase chain reaction-restriction fragment length polymorphism method. RESULTS: The MnSOD genotype frequencies amongst the 150 cases were A/A = 27.3%, A/V = 69.4%, and V/V = 3.3%; the A and V allele frequencies were 62% and 38%, respectively. The MnSOD genotype frequencies amongst the 150 controls were A/A = 33.3%, A/V = 48.0%, and V/V = 18.7%; the A and V allele were 57% and 43%, respectively. We observed a significant difference in genotype distributions of MnSOD A16V polymorphism between patients and controls (p = 0.0001). CONCLUSION: It is suggested that the MnSOD A16V polymorphism may be associated with a risk of female infertility in northern Iran. More studies should be considered with a larger number of patients and controls to confirm our results.

The Relationship between Seminal Melatonin with Sperm Parameters, DNA Fragmentation and Nuclear Maturity in Intra-Cytoplasmic Sperm Injection Candidates.

OBJECTIVE: Melatonin, the chief secretory product of the pineal gland, regulates dynamic physiological adaptations that occur in seasonally breeding mammals as a response to changes in daylight hours. Because of the presence of melatonin in semen and the mem- brane melatonin receptor in spermatozoa, the impact of melatonin on the regulation of male infertility is still questionable. The aim of this study was to determine the effects of endogenous melatonin on human semen parameters (sperm concentration, motility and normal morphology), DNA fragmentation (DF) and nuclear maturity. MATERIALS AND METHODS: In this clinical prospective study, semen samples from 75 infer- tile men were routinely analyzed and assessed for melatonin and total antioxidant capac- ity (TAC) levels using the enzyme-linked immunosorbent assay (ELISA) and colorimetric assay kits, respectively. DF was examined by the sperm chromatin dispersion (SCD) test. Acidic aniline blue staining was used to detect chromatin defects in the sperm nuclei. RESULTS: There was no significant correlation between seminal plasma melatonin and TAC with sperm parameters and nuclear maturity. However, we observed a positive significant correlation between DF and melatonin level (r=0.273, P<0.05). CONCLUSION: Melatonin in seminal plasma is positively correlated with damaged sperm DNA of infertile patients. The mechanism of this phenomenon needs further study.

Oxytocin can decrease germ cells apoptotic index in testis under acute ischemia reperfusion in a rat model.

UNLABELLED: Ischemia reperfusion (IR) is the main pathology of torsion of testis and it is a common urologic emergency. There is some evidence that shows oxytocin (OT) plays role in ischemia reperfusion. OBJECTIVE: To evaluate this hypothesis that OT can decrease germ cell apoptotic index in testis under acute ischemia reperfusion in a rat model. MATERIALS AND METHODS: 20 adult rats were randomly divided into four groups: Control, IR, OT and IR+ OT (OTA). Testicular ischemia was achieved by 720° torsion of the left testis for 2 hr. Then, torsion was removed and reperfusion was performed. Immediately after induction of reperfusion 0.03 µg/kg OT were administered intraperitoneally to the IR+ OT. Three hours after surgery left testis was removed and evaluations were made by Johnson's score, ELISA, immunohistochemistry and histomorphometry for study of maturity of spermatogenesis, endocrine profiles, apoptosis and quantitative studies, respectively. RESULTS: The results showed in addition tissue edema and congestion, a significant reduced in Johnson's score were detected in IR group in comparison with controls (p=0.01), and apoptotic index increased significantly (p=0.001). Administration of OT in OT+IR group, increased Johnson's score but it was not statistically significant. Germinal epithelium thickness was increased significantly (p=0.03), although apoptotic index decreased significantly in comparison with the IR group (p=0.04). However there was not significant difference in serum levels of testosterone, FSH and LH in none of groups (p=0.07). CONCLUSION: These results suggested that OT can decrease apoptotic index and improves complication of acute ischemic reperfusion in testis in a rat model.

The association of ApE1 -656T>G and 1349T>G polymorphisms and idiopathic male infertility risk.

PURPOSE: In spite of variety studies in understanding of human reproductive and fertility, the underlying causes of male infertility remains undefined in about 50 % of cases. The polymorphism studies have a crucial role in diseases recognizing. Human apurinic/apyrimidinic endonuclease 1 (ApE1) is a multifunctional protein that has an important role in the base excision repair pathway. The present study was aimed to evaluate whether two polymorphisms -656T>G and 1349T>G ApE1 are related with the susceptibility to idiopathic male infertility. METHODS: Samples were collected from 180 patients diagnosed with idiopathic male infertility and 120 control subjects and genotyped by tetra-primer amplification refractory mutation system PCR. RESULTS: We observed a significant difference in genotype distributions of -656T>G ApE1 polymorphism between infertile patients and controls (P = 0.0001). Our findings indicated individuals with the variant TG genotypes had a significant increased risk of idiopathic male infertility (OR 1.84, 95 % CI 1.09-3.11, P = 0.021), whereas the significant association between the 1349T>G polymorphism and idiopathic male infertility risk was not observed (P = 0.2). CONCLUSIONS: Our data suggest that the -656T>G ApE1 polymorphism may be associated with increased risk of idiopathic male infertility. Larger studies with more patients and controls are needed to confirm the results.

Vitrification and subsequent in vitro maturation of mouse preantral follicles in presence of growth factors.

OBJECTIVE: Cryopreservation of ovarian tissue or follicles has been proposed as an alternative method for fertility preservation. Although successful vitrification of follicles has been reported in several mammalian species, the survival rate is generally low. The aim of this study was to investigate the effects of fibroblast growth factor (FGF) and epidermal growth factor (EGF) on in vitro preantral follicle development after vitrification. MATERIALS AND METHODS: In this experimental study, preantral follicles with diameter of 150-180 µm were mechanically isolated from ovaries of 18-21 days old NMRI mice. Follicles were vitrified and warmed, then cultured in a-minimal essential medium (α-MEM) without growth factor supplementation as control group (group I), while supplemented with 20 ng/ml FGF (group II), 20 ng/ml EGF (group III), and 20 ng/ml FGF +20 ng/ml EGF (group IV). After 12 days, human chorionic gonadotrophin (hCG)/EGF was added to culture medium, and after 18-20 hours, the presence of cumulus oocyte complexes (COCs) and oocyte maturation were assessed. The chi-square (Χ(2)) test was used to analyze survival and ovulation rates of the follicles. RESULTS: Our results showed that the rate of metaphase II (MII) oocytes in FGF group increased in comparison with control and other treatment groups (p<0.027), but there was no difference between control with EGF and EGF+FGF groups in oocyte maturation rate (p>0.05). There was a significant decrease in survival rate of follicles in EGF+FGE group in comparison with other groups (p<0.008). After in vitro ovulation induction, the follicles in EGF group showed a higher ovulation rate (p<0.008) than those cultured in other groups. CONCLUSION: FGF has beneficial effect on oocyte maturation, and EGF increases COCs number in vitro. Combination of EGF and FGE decreases the number of survived follicles.