Knowledge and beliefs about oocyte cryopreservation for medical and social reasons in female students: a cross-sectional survey.

BACKGROUND: With the increasing number of young women surviving cancer and a growing trend among highly educated women to postpone childbearing for educational or professional pursuits, there is a rising demand for egg freezing services to ensure a successful pregnancy. This study aims to assess the knowledge and beliefs surrounding oocyte cryopreservation, both for medical and social reasons, among female students in Tehran, Iran. METHODS: An online cross-sectional survey was carried out from March to August of 2022, involving a total of 1279 childless students pursuing master's and doctoral degrees at universities in Tehran. The participants were between the ages of 18 and 38. Knowledge and beliefs about medical and social oocyte cryopreservation were assessed through Fertility Preservation Survey (FPS) instrument. RESULTS: The mean age of the participants was 26.38 ± 4.9. The majority of students expected to be "30-34 years" when they become pregnant with their first child (41.1%, M: 30.3 ± 4.13 years) and "35-39 years" when they give birth to their last child (46.7%, M: 35.28 ± 4.18 years). The students agreed with preserving fertility with medical (93.3%) and social (86.9%) indications and believed the medical (95.1%) and social (87.4%) costs of cryopreservation should be covered by the healthcare system. Among the participants, 75.6% considered cost to be a definite or probable factor in their decision to pursue fertility preservation. The oncology team's recommendation was identified as the most important factor in deciding on medical egg freezing (92.6%, M: 3.46 ± 0.71). The overall correct response rate for the knowledge questions was 57.7%. The majority of participants (95.5%) agreed that physicians should routinely provide information about egg freezing to women of childbearing age during their regular healthcare visits. CONCLUSIONS: The research results revealed that female students in Tehran universities have a positive attitude towards medical and social egg freezing, but lack sufficient knowledge about the ideal timing of childbearing. Health professionals could provide detailed information about fertility preservation and age-related infertility as part of routine healthcare visits or reproductive health planning. Additionally, expanding supportive policies and incentives for childbearing established by the government to cover the costs of fertility preservation would be beneficial.

Correction to: "Comparative repair capacity of knee osteochondral defects using regenerated silk fiber scaffolds and fibrin glue with/without autologous chondrocyes during 36 weeks in rabbit model.

The published online of the original version contains mistakes.

Reactive oxygen species-induced alterations in H19-Igf2 methylation patterns, seminal plasma metabolites, and semen quality.

PURPOSE: This study was conducted in order to investigate the effects of reactive oxygen species (ROS) levels on the seminal plasma (SP) metabolite milieu and sperm dysfunction. METHODS: Semen specimens of 151 normozoospermic men were analyzed for ROS by chemiluminescence and classified according to seminal ROS levels [in relative light units (RLU)/s/106 sperm]: group 1 (n = 39): low (ROS < 20), group 2 (n = 38): mild (20 ≤ ROS < 40), group 3 (n = 31): moderate (40 ≤ ROS < 60), and group 4 (n = 43): high (ROS ≥ 60). A comprehensive analysis of SP and semen parameters, including conventional semen characteristics, measurement of total antioxidant capacity (TAC), sperm DNA fragmentation index (DFI), chromatin maturation index (CMI), H19-Igf2 methylation status, and untargeted seminal metabolic profiling using nuclear magnetic resonance spectroscopy (1H-NMR), was carried out. RESULT(S): The methylation status of H19 and Igf2 was significantly different in specimens with high ROS (P < 0.005). Metabolic fingerprinting of these SP samples showed upregulation of trimethylamine N-oxide (P < 0.001) and downregulations of tryptophan (P < 0.05) and tyrosine/tyrosol (P < 0.01). High ROS significantly reduced total sperm motility (P < 0.05), sperm concentration (P < 0.001), and seminal TAC (P < 0.001) but increased CMI and DFI (P < 0.005). ROS levels have a positive correlation with Igf2 methylation (r = 0.19, P < 0.05), DFI (r = 0.40, P < 0.001), CMI (r = 0.39, P < 0.001), and trimethylamine N-oxide (r = 0.45, P < 0.05) and a negative correlation with H19 methylation (r = - 0.20, P < 0.05), tryptophan (r = - 0.45, P < 0.05), sperm motility (r = - 0.20, P < 0.05), sperm viability (r = - 0.23, P < 0.01), and sperm concentration (r = - 0.30, P < 0.001). CONCLUSION(S): Results showed significant correlation between ROS levels and H19-Igf2 gene methylation as well as semen parameters. These findings are critical to identify idiopathic male infertility and its management through assisted reproduction technology (ART).

Ooplasmic transfer in human oocytes: efficacy and concerns in assisted reproduction.

BACKGROUND: Ooplasmic transfer (OT) technique or cytoplasmic transfer is an emerging technique with relative success, having a significant status in assisted reproduction. This technique had effectively paved the way to about 30 healthy births worldwide. Though OT has long been invented, proper evaluation of the efficacy and risks associated with this critical technique has not been explored properly until today. This review thereby put emphasis upon the applications, efficacy and adverse effects of OT techniques in human. MAIN BODY: Available reports published between January 1982 and August 2017 has been reviewed and the impact of OT on assisted reproduction was evaluated. The results consisted of an update on the efficacy and concerns of OT, the debate on mitochondrial heteroplasmy, apoptosis, and risk of genetic and epigenetic alteration. SHORT CONCLUSION: The application of OT technique in humans demands more clarity and further development of this technique may successfully prove its utility as an effective treatment for oocyte incompetence.

Platelet-rich plasma promotes the development of isolated human primordial and primary follicles to the preantral stage.

This study aimed to assess the effects of platelet-rich plasma (PRP) on growth and survival of isolated early human follicles in a three-dimensional culture system. After fresh and vitrified-warmed ovarian tissue was digested, isolated early preantral follicles and ovarian cells were separately encapsulated in 1% alginate (w/v). The encapsulated follicles and ovarian cells were cultured together in a medium supplemented with foetal bovine serum (FBS), PRP, PRP + FBS, or human serum albumin (HSA) for 10 days. Growth and survival of the follicles were assessed by measurement of diameter and staining with trypan blue. Follicular integrity was assessed by histological analysis. After culturing, all follicles increased in size, but growth rate was greater in follicles isolated from fresh samples than those from vitrified-warmed ones (P < 0.001). Similarly, follicular viability of fresh samples after culturing was higher than that of vitrified-warmed ones. The growth and survival rates of follicles from both fresh and vitrified groups cultured in PRP supplemented media were significantly higher than those of other groups (growth P < 0.001 and survival P < 0.05, in both groups). In conclusion, media supplementation with PRP can better support viability and growth of isolated human early preantral follicles in vitro.

Fabrication and characterization of nano-fibrous bilayer composite for skin regeneration application.

Full thickness wound healing with minimal scarring and complete restoration of normal skin properties still remains as a clinical challenge. In this study, a bilayer skin substitute has been fabricated to biomimic the microstructure of natural extracellular matrix of the skin. Human amniotic membrane (HAM) and silk fibroin nano-fibers were combined to produce bilayer construct, which was further treated and characterized. HAM was obtained from healthy mothers and de-epithelized by means of fine enzymatic method to preserve the extracellular structure. Fibroin protein was extracted from fresh Bombyx mori cocoons and transformed to uniform nano-fiberous structure, which was used as a coating layer on the de-epithelized membrane. Surface modification through oxygen plasma treatment was attempted to further induce hydrophilicity. Subsequently, scaffolds were fully characterized in terms of morphology, mechanical properties, hydrophilicity and cell culture response. Histological and immunohistological staining demonstrated localization of fibronectin, cell denudation and structural integrity of HAM after de-epithelization. Scanning electron microscopy images showed bead-free silk fibroin nano-fibers with the average diameter of 250nm. Water contact angle of bilayer scaffolds reduced dramatically to 26.34° after oxygen plasma treatment, which is correlated with more hydrophilic surface. Due to fibroin nano-fiber coating, mechanical properties of HAM improved significantly. Tensile Young's modulus and tensile strength increased from 16.14MPa and 68.46MPa to 25.69MPa and 108.03MPa, respectively. 14days in vitro cultivation of mouse embryonic fibroblasts on the scaffolds revealed that bilayer scaffolds are able to support cell attachment and proliferation. Plasma-etched scaffolds provided the best niche for cell-matrix crosstalk by allowing cells to penetrate beneath the pores and to integrate in fibers direction. The obtained results suggest that the presented nano-fibrous bilayer composite based on HAM is a potential substitute for skin regeneration application.

Comparative repair capacity of knee osteochondral defects using regenerated silk fiber scaffolds and fibrin glue with/without autologous chondrocytes during 36 weeks in rabbit model.

The reconstruction capability of osteochondral (OCD) defects using silk-based scaffolds has been demonstrated in a few studies. However, improvement in the mechanical properties of natural scaffolds is still challengeable. Here, we investigate the in vivo repair capacity of OCD defects using a novel Bombyx mori silk-based composite scaffold with great mechanical properties and porosity during 36 weeks. After evaluation of the in vivo biocompatibility and degradation rate of these scaffolds, we examined the effectiveness of these fabricated scaffolds accompanied with/without autologous chondrocytes in the repair of OCD lesions of rabbit knees after 12 and 36 weeks. Moreover, the efficiency of these scaffolds was compared with fibrin glue (FG) as a natural carrier of chondrocytes using parallel clinical, histopathological and mechanical examinations. The data on subcutaneous implantation in mice showed that the designed scaffolds have a suitable in vivo degradation rate and regenerative capacity. The repair ability of chondrocyte-seeded scaffolds was typically higher than the scaffolds alone. After 36 weeks of implantation, most parts of the defects reconstructed by chondrocytes-seeded silk scaffolds (SFC) were hyaline-like cartilage. However, spontaneous healing and filling with a scaffold alone did not eventuate in typical repair. We could not find significant differences between quantitative histopathological and mechanical data of SFC and FGC. The fabricated constructs consisting of regenerated silk fiber scaffolds and chondrocytes are safe and suitable for in vivo repair of OCD defects and promising for future clinical trial studies.

Development of sheep primordial follicles encapsulated in alginate or in ovarian tissue in fresh and vitrified samples.

In vitro follicle growth is a promising strategy for female fertility preservation. This study was conducted to compare the development of ovine follicles either isolated or in the context of ovarian cortical pieces after short term (8 days) three-dimensional culture in fresh and vitrified samples. Four different experiments were conducted; I) culture of ovarian cortical pieces encapsulated in 0.5% and 1% alginate and without alginate encapsulation (CP-0.5%, CP-1% and CP, respectively), II) culture of isolated primordial and primary follicles encapsulated in 1% and 2% alginate (IF-1% and IF-2%, respectively), III) culture of fresh and vitrified-warmed cortical pieces (F-CP and Vit-CP, respectively), and IV) culture of fresh and vitrified-warmed encapsulated isolated follicles (F-IF and Vit-IF, respectively). The number of secondary follicles after culture was negatively influenced by encapsulation of ovarian cortical pieces (6.3 ± 3.3 and 10.6 ± 0.9 vs 21.5 ± 2.3 in CP-0.5% and CP-1% vs CP, respectively). The diameter of follicles in IF-2% was higher than IF-1% (54.06 ± 2 vs 41.9 ± 1.5) and no significant difference in follicular viability was observed between the two groups. The proportions of different follicular types and their viability after culture in vitrified-warmed cortical pieces were comparable with fresh ones. The viability of vitrified-warmed isolated follicles was lower than fresh counterparts. The growth rate of fresh follicles was higher than vitrified-warmed follicles after culture (47.9 ± 1 vs 44.6 ± 1). In conclusion, while encapsulation of ovarian cortical pieces decreased the follicles' development, it could better support the growth of isolated follicles. Moreover, the viability and growth rate of isolated-encapsulated follicles was decreased by vitrification.

Quantification of sperm specific mRNA transcripts (PRM1, PRM2, and TNP2) in teratozoospermia and normozoospermia: New correlations between mRNA content and morphology of sperm.

Sperm mRNAs could be used as a predictor of fertilization capacity since the transcriptional profile of a gamete is critical for the production of viable human sperm. The aim of this study was to determine if PRM1, PRM2, and TNP2 transcripts in spermatozoa from normozoospermic and teratozoospermic men correlate with sperm morphology and/or assisted-reproduction outcomes. Human ejaculates were collected from 138 men referred to an infertility clinic, and were separated in two groups, teratozoospermic (n =72) and normozoospermic (n =66), based on World Health Organization criteria (2010). Chromomycin A3 and analine blue staining were used to evaluate protamination and chromatin integrity, respectively. Quantitative reverse-transcriptase PCR was performed for PRM1, PRM2, and TNP2. This analysis revealed significantly higher PRM1 and PRM2 mRNA copy numbers in normozoospermic versus teratozoospermic samples (P < 0.001). In contrast, TNP2 transcript abundance was significantly higher in teratozoospermic versus normozoospermic samples (P < 0.001) and positively correlated with sperm-head defects (P < 0.05). Sperm-tail defects negatively correlated (P < 0.05) with both PRM1 and PRM2 transcripts in normozoospermic samples. No significant differences were observed between the two groups when comparing transcript levels to the outcome of intracytoplasmic sperm injection cycles (P > 0.05), and a normal PRM1/PRM2 mRNA ratio (∼1) was observed in more than 70% of successful cycles. Thus, the quantity of PRM1, PRM2, and TNP2 transcripts and the PRM1/PRM2 mRNA ratio affect spermiogenesis, sperm morphology, and the function of mature human sperm. These mRNAs could therefore be used as biomarkers for the diagnosis of male infertility.

Molecular analysis of testis biopsy and semen pellet as complementary methods with histopathological analysis of testis in non-obstructive azoospermia.

PURPOSE: Non-obstructive azoospermia (NOA) is one the many causes of male infertility (10 %) resulting from testicular failure. Multiple testicular biopsies fail to find mature sperm in at least 50 % of cases Therefore; hunting for sensitive and specific biomarkers of spermatogenesis that could better determine the fertility status in NOA can lead to improved management of male infertility. Therefore, we evaluated sperm production through analyses of germ cell-specific transcripts (DAZ, TSPY1, SPTRX3 and SPTRX1) in semen and testicular biopsies of men with azoospermia. METHODS: We collected semen (N=83) and testis biopsies (N=31) from men with non-obstructive azoospermia. We later extracted RNA and synthesized cDNA using washed semen precipitate and testicular tissues. We also performed semi-nested PCR with designed specific primers. Using H&E method, an expert pathologist performed the histopathological evaluation. Having categorized the patients into three groups based on histopathological results, we calculated the agreement between molecular results of semen and tissues with histopathological findings for each patient using Kappa statistical test. RESULTS: Molecular findings of precipitated semen and testicular tissues were in disagreement with histopathological results in most cases. Molecular analysis of testis biopsies showed significant difference (Kappa coefficient=0.009, P value=0.894) with histopathological results; TSPY1, DAZ, SPTRX3 and SPTRX1 were respectively detected in 94 %, 94 %, 17.6 % and 52.9 % of men diagnosed with germ cell aplasia. CONCLUSIONS: Molecular analysis of semen does not provide sufficient sensitivity and specificity to be used as a screening test at the present time, but it is a useful adjunct to histopathological methods in men with NOA. Spermatid/sperm specific transcripts indicated the possibility to find mature sperm following repeated multiple testicular sperm extraction (TESE) or microdisection TESE (mTESE).

Morphologic and proliferative characteristics of goat type A spermatogonia in the presence of different sets of growth factors.

PURPOSE: The present study by using different growth factors was aimed to develop the best practical culture condition for purification of goat undifferentiated SSCs and their colonization under in vitro and in vivo conditions. METHODS: The enzymatically isolated SSCs obtained from one month old goat testes were cultured in DMEM plus FCS supplemented with different sets of growth factors (GDNF, LIF, bFGF, and EGF) for 2 weeks. At the end of each week, the morphological characteristics of cells and colonies alongside with purification rate of undifferentiated type A spermatogonia were evaluated by immunocytochemical staining and flow cytometry. RESULTS: The number and size of colonies in treatment groups were significantly (P < 0.01) higher than corresponding values in control group. In immunocytochemical evaluation, the proportion of KIT and PGP9.5 positive cells were significantly (P < 0.001) higher in control and treatment groups, respectively. CONCLUSIONS: The culture medium comprising all four growth factors, especially the one supplemented with the higher concentration of GDNF, was superior to the other groups with respect to the population of undifferentiated type A spermatogonia and its propagation in culture system. Additionally, goat SSCs could colonize within the mouse testis following xenotransplantation.

Screening for large rearrangements of the RB1 gene in Iranian patients with retinoblastoma using multiplex ligation-dependent probe amplification.

PURPOSE: To screen deletions/duplications of the RB1 gene in a large cohort of Iranian patients using the multiplex ligation-dependent probe amplification (MLPA) technique. METHODS: A total of 121 patients with retinoblastoma, involving 55 unilateral and 66 bilateral or familial retinoblastomas, were included in this study. Among these patients, 121 blood and 43 tissue samples were available. DNA was extracted from the blood and tissue samples and analyzed with an RB1-specific MLPA probe set. The mutation findings were validated with SYBR Green Real-Time PCR. RESULTS: Twenty-two mutations were found in 21 patients; of these, ten mutations were detected in patients with isolated unilateral retinoblastoma. CONCLUSIONS: Our results suggested that MLPA is a fast, reliable, and powerful method for detecting deletions/duplications in patients with retinoblastoma.

Accurate sensitivity of quantum dots for detection of HER2 expression in breast cancer cells and tissues.

Here we introduce novel optical properties and accurate sensitivity of Quantum dot (QD)-based detection system for tracking the breast cancer marker, HER2. QD525 was used to detect HER2 using home-made HER2-specific monoclonal antibodies in fixed and living HER2(+) SKBR-3 cell line and breast cancer tissues. Additionally, we compared fluorescence intensity (FI), photostability and staining index (SI) of QD525 signals at different exposure times and two excitation wavelengths with those of the conventional organic dye, FITC. Labeling signals of QD525 in both fixed and living breast cancer cells and tissue preparations were found to be significantly higher than those of FITC at 460-495 nm excitation wavelengths. Interestingly, when excited at 330-385 nm, the superiority of QD525 was more highlighted with at least 4-5 fold higher FI and SI compared to FITC. Moreover, QDs exhibited exceptional photostability during continuous illumination of cancerous cells and tissues, while FITC signal faded very quickly. QDs can be used as sensitive reporters for in situ detection of tumor markers which in turn could be viewed as a novel approach for early detection of cancers. To take comprehensive advantage of QDs, it is necessary that their optimal excitation wavelength is employed.

Commitment Theory as the Theoretical Framework in Third-Party Reproduction.

Assisted reproductive technology (ART) and third-party reproduction provide the opportunity for infertile couples to have children through different genetic links. This type of treatment has created many challenges for infertile couples. With this treatment, the infertile couple will have a child who is biologically related to the gamete/embryo donor. Accordingly, the transformation that occurs in the structure of traditional families and the concept of parenthood is one of the main consequences and challenges which requires in-depth research. In spite of the successful expansion of infertility treatment and third-party reproduction, there is still no proper social context for implementing third-party infertility treatments in Iran. Therefore, despite the need to use the technology, some couples refuse the treatment unless their confidentiality is preserved. Many couples follow the practice surreptitiously by keeping the donation treatment confidential, to get rid of the existing social stigma and protect their identity, the child and the donor's identity. Commitment theory as a theoretical strategy is proposed to solve the problems of all parties involved in this type of "social and non-biological" parenting. Commitment theory in the context of third-party reproduction expresses the commitment to the contract accepted by the donor and the recipient of the gamete/embryo, based on which, the recipients consider the resulting child as their own, and are committed to all the related paternal-maternal rights and duties such as "alimony" and "inheritance". On the other hand, the donors undertake to waive all their paternal-maternal rights and duties by donating gamete/embryo.

Isolation, identification, and culture of goat spermatogonial stem cells using c-kit and PGP9.5 markers.

INTRODUCTION: Presently the techniques for making transgenic animals are cumbersome, required costly instruments and trained man-power. The ability of spermatogonial stem cells (SSCs) to integrate foreign genes has provided the opportunity for developing alternate methods for generation of transgenic animals. One of the big challenges in this field is development of the methods to identify and purify donor SSCs by antibody mediated cell sorting. PURPOSE: The present study was aimed to identify goat subpopulations of SSCs using polyclonal antibodies against PGP9.5 and c-kit molecular markers as well as the growth characteristics of SSCs during short term culture. METHODS: One month old goats' testicular samples were subjected for immunohistochemical and immunocytochemical evaluations. The enzymatically isolated SSCs were cultured in DMEM plus FCS supplemented with (treatment) or without (control) growth factors (GDNF, LIF, FGF, and EGF) for 2 weeks. At the end of culture the morphological characteristics of SSCs colonies and immunocytochemical staining were evaluated. RESULTS: The number and size of colonies in treatment groups were significantly (P < 0.01) higher than corresponding values in controls. The presence of PGP 9.5 and c-kit antigens was confirmed in immunocytochemical evaluation. In immunocytochemical evaluation, the proportion of c-kit and PGP9.5 positive cells were significantly (P < 0.001) higher in control and treatment groups, respectively. CONCLUSIONS: The presence of PGP9.5 and c-kit antigens was confirmed in goat SSCs. Moreover, culture medium supplementation with growth factors could effectively retain the undifferentiation status of SSCs, reflected as a higher population of PGP9.5 positive cells, after short term culture.

Step-by-step decision-making process in third party assisted reproduction: a qualitative study.

This study explores the experience of decision-making about the third party assisted reproduction in Iranian infertile patients using a qualitative approach. Data were collected in 2017-2019 through in-depth interviews of 20 infertile participants (3 couples and 14 individuals) who became pregnant or had children through third-party reproduction and 12 specialists. The interviews were transcribed verbatim, the data were managed using the MAXQDA 10 and analysed using a conventional content analysis approach. The main category that emerged was 'step-by-step process of adjustment to third party assisted reproduction'. From this, five sub-categories were extracted: (i) 'parental preference for biological children over non-biological children'; (ii) 'parental preference for non-biological children over childlessness threats'; (iii) 'strong preference for having partial genetic link with children'; (iv) 'flexibility in removing or overcoming barriers to third party reproduction'; and (v) 'parental preference for adopted children over childlessness'. The findings in the present study suggest the importance of lineage and biological relatedness in Iranian culture. Patients adjusted to non-biological parenting as a treatment option only if they have to. Using a step-by-step decision-making process helped participants make more thoughtful decisions by organising relevant information and defining alternatives. The results of the present study can help healthcare providers with counselling regarding parenting with or without genetic ties.

The Impact of Third Party Reproduction on Family and Kinship.

The development of in vitro fertilization (IVF) in the UK, in 1978, proved a major breakthrough in the process of human reproduction, which had remained constant in human history. The impact of IVF and the ensuing assisted reproductive technologies (ARTs) has not been limited in revolutionizing the "natural" practice of biological reproduction, but has reached out to and affected almost every institution in society. Family and kinship, as the social expression of reproduction and the institutions which are the most transparently structured realm of human life are those most profoundly affected by ARTs. Although literature on the implications of ARTs is in general abundant, this article presents new insights on their impact on family and kinship in Iran, which remains a unique case in the Muslim world. It explores the particular way ARTs, especially third-party donation, have been endorsed and practiced in Iran, and their consequences for the family, the infertile individuals, and their position vis-à-vis their kin and social group. The conclusion points to the lack of clarity concerning the initial rulings by the Islamic jurists, who allowed the practice of ARTs, and which has led to a number of unintended consequences regarding the legal, religious, cultural, and ethical issues, affecting the family, its structure and the relationship between the kin group. These consequences range, inter alia, from the question of the anonymity of third-party donor, to the permissibility of gamete donation between blood relatives, and to the absence of enforceable legislation.

Testis biopsy in absolute head teratospermia: Is it a logical option?

INTRODUCTION: Absolute head teratospermia (100% abnormal head morphology) is associated with poor fertility and assisted reproductive techniques results. We aimed to find if it is possible to bypass this disorder using sperm retrieved by testis biopsy. METHODS: Multiple testis biopsies were performed in patients with infertility and absolute head teratospermia who were not able to provide semen on the injection day from 2006 to 2018. Then, the found sperms were evaluated based on being proper or not proper for intracytoplasmic sperm injection. RESULTS: Only 2 patients, of a total of 22 (9%), had relatively proper sperms for microinjection. DISCUSSION: There is no benefit to performing testis biopsy in non-azoospermic patients with absolute abnormal head morphology.

Assessing the potential of HSPA2 and ADAM2 as two biomarkers for human sperm selection.

Selection of the best sperm, with the least defects, is a critical factor in the success of ART especially in male factor infertility. This study assessed the potential Heat shock protein (HSPA2) and metallopeptidase domain2 (ADAM2) biomarkers for sperm selection. Sperm were obtained from 72 asthenoteratozoospermic and 42 normospermic ejaculates. The semen characteristic, DNA fragmentation (DFI), chromatin maturation index (CMI), ADAM2 and HSPA2 levels on sperm, and their correlation with embryo quality were assessed in both groups. Results showed the significant reduction in HSPA2 and ADAM2 in asthenoteratozoospermic compared to normazoospermic ejaculates regarding the cut-off value of 14 and 13% for these two biomarkers. The specificity of HSPA2 and ADAM2 separately, and the combination of these two biomarkers, were 95.2, 90.5 and 93.5%, respectively, for sperm from normozoospermic ejaculates. However, they were 48.6, 50.0 and 54.5% for asthenoteratozoospermic ones. A significant correlation was observed with HSPA2, ADAM2 and a combination of these two biomarkers with CMI, DFI and embryo quality. Although a combination of these two biomarkers have the potential to be a good choice for selecting sperm with the lowest level of chromatin damage, it seems that selection according to HSPA2 has priority over ADAM2 or a combination of the two.

Modeling breast acini in tissue culture for detection of malignant phenotype reversion to non-malignant phenotype.

BACKGROUNDS: Evidence is accumulating to support disruption of tissue architecture as a powerful event in tumor formation. For the past four decades, intensive cancer research with the premise of "cancer as a cell based-disease" focused on finding oncogenes or tumor suppressor genes. However, the role of the tissue architecture was neglected. Three dimensional (3D) cell cultures which can recapitulate major aspects of the microenvironment are appropriate models for exploring cancer. For the first time in Iran, we have launched Matrigel based non-malignant, tumorigenic and reverted breast 3D cell cultures. METHODS: Non-tumorigenic MCF-10A and tumorigenic MCF-7 breast cell lines were cultured on plastic and Matrigel. MCF-7 cell lines were reverted to normal phenotype via AIIB2 and LY 294002 inhibitors against beta1 integrin and class I phosphatidylinositol 3-kinase, respectively. RESULTS: MCF-10A acini were distinguishably different from MCF-7 on Matrigel. MCF-10A formed organized hollow spherical structures which were in stark contrast to the MCF-7 disorganized cluster of cells. Matrigel allowed visual monitoring of MCF-7 cells treated with inhibitors. After treatment of MCF-7 cells, we observed reversion of MCF-7 phenotype toward normal, comparable to MCF-10A acini. CONCLUSION: The 3D culture provides a microenvironment which allows malignant and non-malignant cells to demonstrate near physiological behavior and this can distinguish non-malignant from malignant cells. The 3D culture also allows visual monitoring of malignant phenotype reversion to organized spheres.