Cell Journal (Yakhteh)
Year:2009 | Volume:11 | Issue:SUPPL. 1|Papers Count:239

In 1980, the first transgenic mice called the "super mice" capable of producing a human drug, tPA (tissue plasminogen activator to treat blood clots), was produced by the pronuclear microinjection (PNM) technique. However, with the birth of 'Dolly' and 'Polly' in 1997, the methodology of livestock transgenesis was greatly revolutionized. Dolly, of course, is the famous 'cloned sheep' announced in early 1997, and 'Polly' is a transgenic sheep from the same research program. Since then, isolation of human pharmaceutical proteins from milk of genetically modified animals became a priority for all the countries. Accordingly the cloning animal using the delicate technology of somatic cell nuclear transfer (SCNT) has expanded from handful research laboratories in to over 200 laboratories across at least 40 nations.This perspective describes a historical background of the current status of cloning and transgenesis technology in Iran regarding the worldwide’s developments. We outline the experimental designs, the pitfalls and challenges encountered, and the eventual success in developing a practical approach. Finally, our suggestions of the promises of these two tightly joined technologies for advancing basic research science and pharmaceutical and therapeutic strategies will be discussed.

In mammals, the initiation of ovarian follicular growth occurs as the oocyte enlarges and the immediately adjacent somatic (i.e. granulosa cells) develop a uniformly cuboidal appearance and proliferate. Whilst this transformation is accompanied by the expression of several hundred new genes not expressed in the primordial follicles, the key factors responsible for the initiation of growth are not well understood. Nevertheless, it has now been established that the oocyte plays an essential role in regulating follicular growth once growth has been initiated, at least until follicles develop a dependency on the pituitary hormones, LH and FSH for preovulatory maturation. During the period preceding gonadotrophin dependence, the oocyte is thought to regulate follicular growth through the secretions of growth factors such as bone morphogenetic protein (BMP)-15 and growth and differentiation factor (GDF)-9 and by interactions with the granulosa cells via an extensive network of gap-junctions. In this way, the oocyte is thought to regulate the proliferation and differentiation of its adjacent granulosa cells into two distinct phenotypes, namely, the cumulus cells (those immediately adjacent to the oocyte) and the mural granulosa cells (those more distantly located from the oocyte). Moreover, from studies in sheep, it appears that the localised concentrations of BMP-15 within the developing follicle influences the timing of formation of receptors for LH on mural granulosa cells and thus the number of follicles that may go on to ovulate. Currently, there is intense interest in determining the molecular forms and mechanisms of action of BMP-15 and GDF-9 and the potential applications of these growth factors for improving fertility in vivo and for use with in vitro technologies to improve IVF outcomes.

Studies of the genetics of ovulation-rate have proven helpful for understanding the physiology of ovarian follicular function in mammals. Since 2000, studies of the inherited patterns of ovulation-rate in sheep have revealed at least 6 different point mutations in either the pro- or mature protein regions of bone morphogenetic protein (BMP)-15 and at least two different point mutations in the mature protein region of growth and differentiation factor (GDF)-9. Animals homozygous for any of the X-chromosome derived BMP-15 mutations (e.g. FecXB) and one of the GDF-9 mutations (FecGH) on sheep chromosome 5 are sterile with streak ovaries, whereas, those heterozygous have ovulation-rates approximately one higher than those of the wild-types. Animals heterozygous for both FecGH and FecXB have ovulation-rates that are additive for each mutation separately indicating that these growth factors can co-operate with one another. Within the sheep ovary, both BMP15 and GDF9 are expressed exclusively by oocytes. These findings together with those from other studies in cattle and humans show that the oocyte has a major role to play in regulating ovulation-rate in mammals. In addition to BMP15 and GDF9, a point mutation in a BMP type I receptor known as activin-like kinase (ALK)-6, on sheep chromosome 6, also has a profound influence on ovulation-rate. Animals heterozygous and homozygous for the ALK-6 mutation have ovulation-rates that are 1.5- and 3.5-fold higher respectively, than their wild-types. As with BMP-15 and GDF-9, the ALK-6 gene is expressed in oocytes, but also in granulosa cells of developing ovarian follicles. Other mutations in sheep, known to effect ovarian follicular development and/or ovulation-rate, have been localised to chromosomes 5 (Thoka ewes), 11 (Lacaune) and X (Woodlands), but these genes remain to be identified. In the case of the Woodlands mutation (FecX2W), it seems that this also affects ovulation-rate at the level of the ovary as the patterns of expression of the BMP-15, ALK-6 and ALK-5 genes in oocytes or granulosa cells differ between those carrying the FecX2W mutation and the wild-type.In summary, a remarkable range of naturally-occurring ovulation-rates varying between 0 and 15 have been identified in domesticated sheep. All the genes identified as responsible for this diversity are expressed in oocytes and/or adjacent follicular cells. An understanding of how these factors influence follicular development, ovulation rate and fecundity are currently the subject of intense research.

Cryostorage and grafting of frozen-thawed (F/T) isolated ovarian follicles, ovarian tissue fragments or whole ovaries are all possible therapeutic approaches now available to preserve fertility in young patients undergoing potentially gonadotoxic radio/chemotherapic treatments for cancer or other systemic pathologies. Our aim was to evaluate through a morphological approach the impact of enzymatic isolation, cryopreservation and grafting on ovarian tissue integrity and viability.  Human ovarian tissue was analyzed after the following treatments: 1- Follicle isolation by collagenase or Liberase enzymatic digestion; 2 - Xenotransplantation and orthotopic autotransplantation of F/T ovarian cortical strips; 3 - Cryopreservation of intact ovaries with their vascular pedicles. Observations were carried out by transmission electron microscopy, vital fluorescent staining, immunohistochemistry and DNA strand breaks analysis by TUNEL. In treatment 1, a higher proportion of follicles were viable after Liberase isolation in respect to those isolated with collagenase, and most of Liberase-treated follicles were of good ultrastructural morphology. These data imply that the treatment with Liberase - a purified, endotoxin-free enzyme blend - is a promising alternative to impure collagenase preparation for the isolation of intact ovarian follicles for culture and grafting purposes. In treatment 2, cryopreservation and transplantation do not appear to greatly affect the morphology of human primordial/primary follicles, which even grow in the grafts; however, follicular density was reduced after transplantation and follicular development appeared initially impaired. Thus, further studies are needed to extend follicular life span and to improve follicular growth in the graft. In treatment 3, cryopreservation was not associated with any particular sign of apopotosis or ultrastructural alteration in all ovarian (follicular, vascular and stromal) compartments. This result suggests that whole-organ sampling and transplantation may be a viable option in the future, since vascular pedicle anastomosis allows immediate revascularization and greatly reduces ischemic injuries in the transplanted tissue.Funds were provided by Italian Ministry of Education, University and Research (university grants) and by Italian Ministry of Foreign Affairs.

In the field of assisted reproduction technology oocyte cryopreservation protocols have not been fully optimized yet and overall clinical success remains relatively suboptimal. Further experimental (morphological, biochemical, biomolecular) and clinical studies seem thus necessary, in order to better comprehend the effects on human oocytes of all factors associated with freezing and ultimately tailor the best protocol for human oocyte cryopreservation. Our aims were to evaluate the ultrastructure of human preovulatory oocytes frozen-thawed (F/T) using different protocols of cryopreservation and to compare the data obtained with the morphology usually shown by fresh (control) oocytes. In detail, our study investigated the ultrastructural features of oocytes subjected to various procedures of slow cooling and vitrification. Particular emphasis has been given on how the vitrification process impacts the ultrastructural morphology of the mature oocyte. In fact, vitrification currently offers interesting perspectives in the field of oocyte cryopreservation, being judged more tolerable than slow cooling by the oocyte. Despite this, very little is known about the fine morphology of vitrified-warmed oocytes, and the available studies are almost totally concerning non-human species. F/T oocytes were vitrified using two alternative devices (cryoloop and cryoleaf). By light microscopy both fresh and F/T oocytes were generally rounded, 90-100 microns in diameter, provided with an ooplasm showing a uniform texture and surrounded by a continuous zona pellucida. Sparse microvacuolization was only occasionally detected in both fresh and F/T oocytes, at the same extent. Transmission electron microscopy (TEM) evaluation confirmed the very sporadic incidence of ooplasmic vacuolization in fresh and F/T oocytes. By TEM, oocyte organelles mostly consisted of mitochondria-smooth endoplasmic reticulum (M-SER) aggregates, varying in shape, size and location. In particular, in more than a half of the F/T oocytes observed, M-SER aggregates appeared slender in shape and smaller in size than those observed in fresh oocytes; in spite of this, a proper mitochondrial fine structure was generally maintained in all these samples. In the remaining F/T oocytes, and in fresh oocytes as well, both small and large aggregates were found randomly distributed in the ooplasm. Finally, numerous F/T oocytes displayed a non-homogeneous distribution of microvilli and a reduced complement of cortical granules (CGs) – the latter particularly evident in those oocytes vitrified using the cryoloop device. In conclusion, a) the virtual absence of ooplasmic microvacuolization seems the most relevant marker of quality in vitrified-warmed oocytes, irrespective of the device utilized; b) CG pattern appears better preserved in cryoleaf-supported oocytes; c) the finding of underdeveloped M-SER aggregates and altered microvilli emphasize the need of further ultrastructural studies on human vitrified oocytes.Funds were provided by Italian Ministry of Education, University and Research (university grants) and by the Italian National Health Institute.

Surrogacy motherhood, as a method of assisted reproduction, is being widely practiced in Iran. The acceptance and legitimacy of this practice, in the absence of pertinent rules and regulations, is according to the Jurisprudential Decrees (Fatwas) obtained from some of the Shiite religious authorities. According to these Fatwas, commercial surrogacy is being practiced in Iran. Commercial surrogacy, in contrary to the altruistic version, seems to be in conflict with the very concept of human dignity.Human dignity, as a notion rooted in both secular and religious schools of thought, can be regarded as a common platform of ideas among the main existing schools of moral philosophy and ethics. This concept can be relied on, for reaching to common principles and norms among all these schools.According to the Shiite theology, notions such as human dignity could have independent logical basis regardless of their roots in the Holy Scripture. Therefore, human dignity can be considered as a concept which is found by reason and emphasized by the scripture. However, it seems that in the current jurisprudential discourse among religious scholars, when the bioethical issues are being discussed, more attention and observation to this principle is needed.In this article, after a brief situation analysis of the current practice of surrogacy in Iran, I will assess the jurisprudential and ethical views toward the altruistic and commercial surrogacy and will conclude that although the altruistic one is acceptable by both Shiite jurisprudence and medical ethics, the commercial surrogacy, because of its conflict with human dignity, arises some major ethical concerns and a revision of jurisprudential decrees in this regard is necessary.Also, I will argue that surrogate motherhood can be discussed as an example showing the necessity of the observance of the concept of human dignity in the future of Islamic bioethical jurisprudence.  

Semen donation is one of the earliest - and technologically least sophisticated - forms of assisted reproduction, with evidence of physicians offering the service at least as far back as the mid 19th century CE. However, from the outset, the dubious acceptability, morality and legality of the practice ensured that it was practiced as a clandestine rather than simply a confidential medical service. Physicians ensured that donors (frequently their friends and professional colleagues) and recipients never knew of each other’s identity, often advised recipients and their partner not to tell anyone – including their child(ren) – of their recourse to semen donation, and no provisions were made for donor-conceived people to learn anything about their donor. Secrecy, anonymity and donor recruitment practices promoted the use of semen from a single donor on multiple occasions, and it is known that some donors have helped to conceive many dozens of offspring. As demand for semen donation increased, physicians sought to recruit new donors, offering modest financial reward as an incentive. Subsequent technological developments enabled oocyte and embryo donation to become widely available, applying similar practice models to those already adopted for semen donation, although higher levels of financial compensation have tended to be offered to oocyte donors. While semen, oocyte and embryo donation were initially intended to assist heterosexual couples with a fertility difficulty to conceive a child, they have become a key means by which single women and lesbian couples (using donated semen) and post-menopausal women (using donated oocytes or embryos).Although comparatively little empirical information concerning gamete donation is available, primarily because it was set up in a way that precludes the accumulation of an adequate evidence base, a number of ethical issues have emerged and which will be discussed in this presentation, including:• Service providers’ responsibilities to promote evidence based practice• Service providers’ responsibilities towards donors, recipients and donor-conceived individuals• Welfare of donor-conceived individuals-including • Separation of children from their genetic parent(s)• Limits on the number of offspring conceived by the gametes of a single donor• Information for donor-conceived individuals about their genetic and biographical history and about their genetic relatives • Use of gamete and embryo donation by single women, individuals in same sex relationships and older, post-menopausal women.• Donors’ responsibilities and obligations • Commercialisation of gamete procurement and financial rewards for donors • Risks of donor exploitation

Over 3.5 million children are believed to have been born worldwide – and over 200,000 annually - as a result of assisted reproduction procedures. Such procedures account for an increasing proportion of all births; for example, among European countries where data are available, approximately 1.7% of all births each year result from assisted reproduction procedures.It is commonly asserted that the welfare or interests of children born as a result of assisted reproduction procedures should be a high priority of service providers - a requirement that is enshrined in statute in some jurisdictions. Such requirements have tended to focus exclusively on the suitability of potential parents of children conceived through assisted reproduction procedures; this focus has highlighted the difficulties of operationalising both welfare principles and requirements.However, the impact of specific assisted reproduction procedures on children has received less attention, and while it is frequently claimed that assisted reproduction procedures have become routine practices, the underpinning evidence base, especially regarding the long-term safety of assisted reproduction procedures, is less robust than for other similarly well-established procedures.This presentation will highlight these issues by drawing on the specific examples of intracytoplasmic sperm injection (ICSI), multiple births, pre-implantation genetic diagnosis (PGD), selecting the characteristics of children, and use of previously cryopreserved embryos, oocytes, and semen.The presentation will identify key challenges that need to be addressed to further promote the welfare of children conceived through assisted reproductive procedures:• Accurate information and comprehensive national registration systems• Further research to identify longer-term health, psychological and social implications of assisted reproductive procedures on children and their families • Establishment of formal requirements to take account of the welfare of children conceived through assisted reproductive procedures.

Posthumous assisted reproduction is the most challenging, difficult and sensitive issue to be discussed ethically and religiously. In this paper the acceptability of the posthumous reproduction in Islamic contexts is evaluated and major concerns like Consent and ownership of the gametes after death, Family and Marriage vision and Welfare of the child are discussed together with some international legislation. We can conclude that upon Islamic vision to assisted reproductive techniques as treatment of families and relieving the serious problem of childlessness, posthumous assisted reproduction is unacceptable for the remaining partner even with previously frozen gametes or embryos. Also, Islamic vision to marriage, consent and welfare of the child confirms the unacceptability. Posthumous gamete or embryo donation is different. The frozen embryos or gamtes after the person’s death can save another infertile family with less problems. As the parent is dead, upon current law, after one year, there is no lineage related to the child who is born. So, problems like intimacy, inheritance, expenditure and guardianship will be solved. It seems before starting such a program, there is an extreme need for a complete research and obtaining decrees and pursue a complete guideline preventing any future problem.

Biological markers are material measures obtained from the bodies or excretions of individuals potentially usable to detect: environmental exposures, effects of exposures.The rationale for biological markers in epidemiologic research is strong in that markers have the potential for improving the accuracy of our "exposure variables", permitting the identification of preclinical disease and providing opportunities for prevention, allowing for more homogeneous and etiologically relevant classifications of disease, and enhancing our understanding of the biological processes leading to disease occurrence.When we are dealing with biomarkers, they need validation using laboratory, epidemiologic, and clinical studies. The analytic validity, clinical validity and clinical utility of a biomarker are important for evaluating it. The use of biomarkers in epidemiologic research raises a number of ethical, legal, and social issues.Biomarkers in reproduction epidemiology are considered as markers of events before and around fertilization, biological markers in discovering or interpreting female reproductive disorders that might be owed to environmental causes, markers of the pre- and peri-implantation phases and markers of the postimplantation phase, experience with studies of chromosome anomaly in spontaneous abortion.

The advent of any new scientific development that have wide applications and which impinge on human life raise several technical and moral dilemmas and poses many ethical and technical challenges. The assisted reproductive technology (ART) is no exception. ART also raises questions from society on their ethics and safety; in some instance moral issues also are raised. An additional factor arises when technology becomes privatized, with a possible loss of Govt. control. Unlike most other medical techniques, success rate using ART is poor. However, the desire of those whose marriages have remained barren, to have children, specially in our country, is so great that many infertility clinics with little expertise or reliability have come up all over the country. The services offered by some of these clinics are questionable. There is no formal training in this discipline in the country. The mushrooming of infertility clinics in India has been, therefore, a matter of great concern. This in turn creates pressure on politicians and legislators to scrutinize understand and control these practices. The reasons for this state of affair is the lack of National Guidelines and an Accreditation, Supervision and Regulatory body under which all infertility clinics offering ART would be placed. The Indian Council of Medical Research (ICMR) had developed "National Guidelines for Accreditation, Supervision and Regulation of ART Clinics in India" in consultation with the National Academy of Medical Sciences (NAMS) (India). The draft guidelines were widely publicized discussed and debated by expert groups of the ICMR and then by practitioners of ART and the public throughout the Country. The National Human Rights Commission (NHRC) and National Commission for Women (NCW) have also approved the guidelines. The final guidelines have also been accepted by the Ministry of Health and Family Welfare, Govt. of India. To implement these guidelines in the country the guidelines have been translated into draft Assisted Reproductive Technologies (Regulation) Bill & Rules -2008 which was also subjected to public debate. Based on the comments & suggestions of various stake holders, both nationally and internationally and after consultation & approval of the experts the draft ART (Regulation) Bill & Rules has been revised and after the approval of the respective Ministries, Govt. of India it will be placed in the parliament of Republic of India for approval.Artificial insemination by sperm (AID) has long been used to achieve successful pregnancies when the male partner is unable to father a child, or when there is a danger of transmitting a hereditary disorder to the offspring. After the development of IVF, the donation of eggs and embryos has also become a possibility. There are clear ethical parallels between sperm and egg donations. Like sperm, donor eggs can help infertile individuals achieve a pregnancy and have a child who is genetically related to one parent. In the case of Donor embryos, it is enabling a woman to go through pregnancy and childbirth. Even in surrogacy the child genetically belongs to either both the parents or at least one parent. However for some couples sperm or eggs might be thought to raise problems in so far as the social father or mother may feel less attached to a child that is not genetically his/her. This problem could be compounded by the use of donor embryos in few cases. But such problems should largely be able to be overcome by adequate counseling prior to the use of donor gametes or embryos.One of the central ethical questions relating to egg, sperm and embryo donations is whether a child conceived in this way should or should not be told to her or his origin and what would happen in each case. Traditionally, as also in past, it was the almost universal practice to keep the identity of both the sperm donor and the recipient confidential. This practice has been challenged on the grounds that the child has a “right to know the truth”, or that it is too difficult to hide the truth. Several countries now allow children when they reach majority to access at least non-identifying information about their genetic parents. Surrogacy is another important issue as infertile couples from different parts of the world comes to India for hiring Surrogate mother. There are number of issues both nationally and internationally in engaging surrogate mother and needs proper attention. Appropriate measures have been recommended in the Councils guidelines for ART Clinics in India as well as in draft Assisted Reproductive Technologies (Regulation) Bill & Rules -2009 on various issues related to AID, ovum donation & surrogate mother and will be discussed during the presentation.

Recent advances in reproductive technologies have transformed the way people look at having children. Childless couples need not, and do not perceive their life a cursed one, thanks to the increasing practice of donor insemination, egg donation, in vitro fertilization, embryo transfer, surrogacy and its acceptance. Surrogacy is now a viable clinical option for childless couples. The two practices Egg donation and Surrogacy or contract pregnancy as some prefer to call it, raise very different ethical issues along with payment for Surrogacy.It has become evident that it is generally the socio-economically marginalized women who agree to act as surrogates due to the financial benefit it entails. This not only puts these already-vulnerable women in situations where their bodies may be exploited for the benefit of other people, but also jeopardizes their physical and mental health, thus making them ‘objects of reproduction’. Donors should not be paid for their eggs, but rather they should be compensated for the burdens of egg retrieval. Making the distinction between compensation for burdens and payment for a product has the advantages of limiting payment, not distinguishing between donors on the basis of their traits, and ensuring that donors are paid regardless of the number or quality of eggs retrieved.India is a hot destination for surrogacy because not only does it have a lot of successful IVF Surrogacy patients and good doctors there are a lot of women who are ready to be surrogates. The cost is the most beautiful part. Where the process takes about $60,000 in U.S, it is done is as much as $3000 in India. Who won’t take this offer up? Who won’t want to have a child in their arms after trying for years?

The right to reproduction is one of the fundamental innate human rights.Surrogacy is the only way to overcome both biological and the social infertility, the only chance to have a child of one’s own for millions of people in the world. Unjust and illogical bans deny people this right and lead to the so-called reproductive tourism.The legal status of surrogacy and approach to this issue vary from one country to another. In a number of countries, such as Germany, France, Italy, Switzerland and Austria, surrogacy is prohibited by law. Other countries, such as Great Britain, Canada and certain Australian states allow surrogacy programmes only on a non-commercial basis, only the expenses linked to the surrogate mother can be reimbursed. Still other countries, like Russia, Ukraine, Belarus, Georgia, Moldova and a number of US states allow surrogacy on a commercial basis. Yet, in other countries, like Belgium, the issue of surrogacy is not stipulated by law, and the woman who gives birth to the child automatically becomes its mother.In some Islamic countries surrogacy is not prohibited by law, yet it is not admissible from the point of view of the religious authorities.

Since the beginning of the human era, history has registered birth of the so-called ‘posthumous’ children. Most jurisdictions recognized these children as legal for a certain period of time after their fathers’ death.The breakthrough of assisted reproductive technologies has led to the development of extraordinary possibilities. Now a child can be born many years following the death of one or both of its biological parents.‘Posthumous’ programmes are implemented through the usage of the cryopreserved material, often accompanying surrogacy and donation programmes. That is the scenario according to which in 2005 the first Russian ‘posthumous’ reproductive programme was realized for the Zakharovs family, who used the cryopreserved sperm of an unmarried young man who had died nine years before and a donor’s oocytes. The child was carried be a‘gestational’ surrogate mother.The posthumous reproduction manifests itself as the realization of the human right to reproduction even though after the death and the child’s right to be born. Posthumous sperm collection is carried out by certain countries and has no legal ground, presuming that any human being would like to become a parent.Today none of the countries in the world have a distinct legal stipulation for the implementation of such programmes, nor have they established the principles of registration of ‘posthumous' children and protection of their legal and property rights.

Women affected with polycystic ovarian syndrome (PCOS) are the most difficult patients for a successful treatment with controlled ovarian hyper stimulation (COH). Their multiple and small antral follicles may be either resistant or highly sensitive to gonadotropin stimulation. Numerous controversies exist concerning the best protocols for folliculogenesis in PCOS patients.One new approach for ovarian stimulation, which has been reported in some previous studies, is adding low-dose human chorionic gonadotropin (hCG) in the late follicular phase. The role of LH in sensitizing antral follicles to FSH is unclear. LH is required for normal hormone production and normal oocyte and embryo development, but follicular responses to LH may depend upon the stage of development. Low-dose hCG has a longer half-life and lower cost compared to recombinant LH. In addition, full development of large follicles, adequate ovarian hormonal level, oocyte maturation, avoidance of premature LH surge, and increment of pregnancy have been demonstrated by adding of low-dose hCG in late folliculogenesis.Recently, Filicori et al showed that the completion of folliculogenesis could be achieved with the administration of low-dose human chorionic gonadotropin (hCG) in controlled ovarian hyper stimulation after 7 days of FSH-only priming. They have also reported a similar pregnancy rate with the use of this stimulation protocol in in vitro fertilization (IVF) with intracytophasmic sperm injection (ICSI). Traditionally, the role of LH in the control of the menstrual cycle was believed to be limited to stimulating theca cell androgen production, triggering ovulation and support of the corpus luteum. However, the physiologic selection of the dominant follicle in spontaneous menstrual cycles is now believed to be the result from the expression of LH receptors in the more mature ovarian follicles (>10 mm in diameter).Ashrafi et al in their recent RCT compared the efficacy of two regimens of low-dose hCG (100 IU/ day or 200 IU/ day for folliculogenesis of PCOS women. These authors could not find any significant differences about stimulation duration and mean number of mature oocytes. However, immature oocytes were significantly lower in patients who received 100 IU hCG (group B). Study groups (groups B, C) also had lower gonadotropin consumption than the control group. Fertilization, implantation, and pregnancy rates were similar amongst the groups.In summary, it seems that this new ovarian stimulation protocol permits follicles and oocytes to fully develop; helps generate top-quality embryos, avoids premature ovulation, establishes clinical pregnancies, reduces administration of recombinant hFSH, minimizes costs, and does not increase the chances of OHSS.

Stress is a generic term for a state of psychosocial disharmony and connotes a panopoly of meanings. Physiologically, stress is understood as activation of processes in response to events and conditions. Acute stress is associated with brief behavioral challenges that elicit adaptive neuroendocrinologic responses termed “homeostatic” that promote short-term survival. Chronic stress is associated with prolonged behavioral challenges. The neuroendocrinologic adaptations associated with chronic stress and protracted behavioral challenge are termed “allostatic.” Allostasis is the achievement of stability through change and entails the alteration of the feedback “set point” for a given neuroendocrine arc.Reproductive “alignment” occurs when physiological responses elicited by the external milieu modulate reproductive processes. A variety of factors serve as determinants and mediators of reproductive alignment. Because factors interact, the independent impact of any one factor may be minimal and difficult to discern. Indeed, a constellation of factors determines whether reproduction is promoted or impaired. Reproductive processes subject to environmental modulation include ovulation, fertilization, embryogenesis, implantation, placentation, and parturition. Clinical manifestations of reproductive misalignment include infertility, recurrent miscarriage, intrauterine growth restriction, preterm labor, low birth weight, preeclampsia, and congenital birth defects. Mechanisms mediating these outcomes include alteration of physiological patterns of the hypothalamic-pituitary-adrenal (HPA) and -thyroidal (HPT) axes and alteration of genetic patterns, such as methylation of DNA.  Physiological processes are generally viewed as plastic and reversible. While epigenetic alterations may be to some extent reversible, they also may be generationally transmitted. The term “fetal origins of adult disease” connotes disease or disorders due to transmissible epigenetic imprinting and underscored the notion that health promotion and predictive health begins before birth.Not all maternal or paternal conditions that can compromise fetal well-being are readily recognized and include endocrine conditions such as subclinical hypothyroidism, impaired glucose tolerance, and diabetes, autoimmunity, infections, and genetic status.  Stress is often an unappreciated cause of reproductive compromise and is difficult to recognize. While stress is generally categorized as metabolic or psychogenic, strictly speaking, these are not separable entities. Behaviors that activate stress signaling pathways activate pathways subserving metabolic and psychogenic outputs. Chronic activation of the HPA and/or suppression of the HPT can impede hypothalamic-gonadal function in both men and women. Individuals with functional hypothalamic hypogonadism typically display a combination of behaviors in response to ongoing psychogenic challenge that concomitantly induce mild energy imbalance and, in the context of chronic adrenal activation, increase the metabolic costs of common activities including recreational exercise.Stress-induced anovulation (SIA), also referred to as functional hypothalamic amenorrhea (FHA), provides a classic and overt example of reproductive compromise and is characterized by the cessation of menses and infertility. SIA is accompanied by a constellation of neuroendocrine secretory aberrations and psychological correlates that include unrealistic expectations of self and others, perfectionism, and maladaptive attitudes about body image and food. However SIA represents only a small portion of the possible presentations for reduced gonadal function in women. Other, less overt, forms of reproductive compromise are far more prevalent and include polymenorrhea (menstrual interval < 24 days), oligomenorrhea (menstrual interval > 35 days), and luteal insufficiency with preserved menstrual interval. Occult reproductive compromise may only come to clinical attention if fertility is desired, especially in men.While it is generally appreciated that maternal health impacts fetal health, the exact mechanisms mediating this link remain to be fully explicated. One must also recognize that factors with the potential to influence reproductive processes are not limited to those contributed by the parents. Medical interventions intended to facilitate reproduction also have the potential to activate these mechanisms. Thus, while assisted reproductive technologies allow us to bypass many of the mechanisms that mediate reproductive compromise, they also alter the developmental milieu during fertilization and early embryogenesis.

Multiple mechanisms mediate the impact of hormones upon brain. Brain functions putatively impacted by hormones fall into categories such as motor, mood, and cognition. Hormones also have been implicated as hastening, protecting, or having nil effect upon molecular processes subserving neurodegeneration. Due to insufficient and contradictory findings, the debate over the importance of hormones for brain health continues unabated and clinicians and patients alike are left with few solid guidelines. Indeed, awareness that the brain is an important target tissue of steroids is still emerging. To complicate matters, drugs which modify or alter hormone levels have been added to the clinical armamentarium, but rarely has the impact of these hormonal mediators upon the brain been studied at molecular and clinical levels. Hence, drugs which modify or interdict hormone action are now in relatively widespread use while belatedly we are trying to understand the neurological, psychological, behavioral, and psychiatric implications.Recently, we have focused on the impact of hormones upon brain neurotransmission using state-of-the-art neuroimaging approaches. In parallel, we have explored the impact of SERM’s and AI’s upon cognitive measures in women being treated for breast cancer. Both approaches confirm that hormones play a significant role in modifying neural substrates subserving cognition and emotion. In particular, it appears that aromatase inhibitors impart a more deleterious impact upon cognition than does tamoxifen in women being treated for early stage breast cancer. Further, serotonergic neurotransmission, which subserves cognition, mood, metabolism, and vegetative activities such as sleep, is modulated by physiological concentrations of estradiol and progesterone in young postmenopausal women. When considered in the context of epidemiological studies and other information about the molecular, cellular, and physiological functions of hormones, one must conclude that future studies that intend to delineate the pros and cons of hormonal modifiers should include the brain as one of the important target tissues. In particular, comparative neuroimaging studies are suggested as a means of determining which estrogenic substances might best support the postmenopausal brain.

With the advent of increasing molecular characterization of individual differences, it has become ever more important to understand how we might wisely individualize our approach to hormone use. The need to individualize, in turn, has spurred an interest in determining and understanding more about how estrogens differ. From the clinical point of view, we know that not all women are the same and we know that not all estrogens are the same. But which of the differences between women or hormones are the ones that really matter when making clinical decisions about birth control or menopause management?When transdermal estradiol formulations for menopause management became available in the early 1990s, the dogma that all estrogens were equivalent was called into question. It was recognized that the transdermal route of administration of estradiol had less hepatic impact than the oral route and that this might confer benefits, including a reduced risk of venous thromboembolism. This line of reasoning seemed to make sense until a recent report (Cole et al. Obstet Gynecol 2007; 109: 339-346) showed that the transdermal contraceptive patch leads to higher rates of venous thromboembolism than oral contraceptives. Taken together, these observations beg the question as to whether route of administration of estrogens does indeed matter.To dissect these findings, we must consider the pharmacologic attributes of different estrogen preparations and the physiologic mechanisms that are influenced by type, route of administration, and dosage of estrogen formulations. The goal is to illustrate the concept that not estrogens are the same and to highlight our knowledge gaps. We will likely be afforded a more clinically relevant perspective about the clinical differences between estrogens that are more chemically similar as we learn more about how each differentially harnesses intracellular machinery, such as protein chaperones.

Recurrent miscarriage is often defined as three or more consecutive miscarriages. The incidence of recurrent miscarriage among couples trying to conceive is 1–3%. Recurrent pregnancy loss may be a consequence of an abnormal embryonic karyotype, or maternal factors affecting the endometrium resulting in defective implantation. The causes for repeated implantation failure may be because of reduced endometrial receptivity. Various uterine pathologies, such as thin endometrium, endometriosis, hydrosalpinges, suboptimal ovarian stimulation, altered expression of adhesive molecules and immunological factors, may decrease endometrial receptivity.In order to study the endometrial factors responsible for recurrent pregnancy loss, endometrial biopsy samples should be precisely timed according to the LH surge, and the investigation should be carried out in a non-conception cycle, prior to the next pregnancy. The various methods of studying the endometrium including morphological studies, morphometry, immunohistochemistry, measurement of endometrial protein in plasma and uterine flushings, cytokine expression in endometrial cells, leukocyte populations in the endometrium will be discussed.

Objective: In this review article the evidence based data about the late complications of PCOS will be discussed. Lifestyle and exercise and management are the important points in control and prevention of the sequels.Materials and Methods: Medline and Cochrane review.Results: Polycystic ovary syndrome is associated with a variety of clinical disturbances including infertility, menstrual dysfunction, hyperandrogenism , elevated risk of pregnancy complications, and metabolic abnormalities such as increase in the prevalence of obesity, insulin resistance, and compensatory hyperinsulinemia, endometrial cancer and cardiovascular diseases. The problem of overweight and obesity is extremely prevalent in the PCOS population, and there is considerable evidence that obesity plays a negative role in the pathophysiology of PCOS. Lifestyle and genetics are the common predisposing factors in PCOS. The majority of studies support the positive effect of weight loss on improving risk factors for CVD and T2DM, reproductive endocrine parameter and outcomes in PCOS. In this lecture the role of exercise and diet in treating the syndrome, which though consistently recommended as the frontline therapy will be discussed. Lifestyle modification programs with an emphasis on behavioral management and dietary and exercise interventions have been successful in reducing the risk of diabetes and the metabolic syndrome in the general population and improving reproductive and metabolic features in PCOS. There is limited evidence for specific dietary and exercise approaches and guidelines for use in PCOS.These strategies can be implemented into longer-term weight maintenance regimens through use of lifestyle modification techniques that consist of a multifaceted approach of dietary, exercise, and behavioral therapies that aim to educate the individual with principles and techniques to achieve dietary and exercise. Alternatives to lifestyle management are anti-obesity pharmacology drugs such as metformin and bariatric surgery that they have limited and short time benefits for weight loss and lifestyle modification.Conclusion: the most important aim of PCOS management is control and prevention of late onset complications and lifestyle modification and secrecies have proved to be critical in the management of PCOS.

Ovarian hyperstimulation syndrome (OHSS) is an iatrogenic and life threatening syndrome with abdominal pain, nausea, vomiting, bloating, ascitis and sometimes with end organ damage resulting from excessive ovarian stimulation. The reported incidence of OHSS varies from 1% to 10% of in vitro fertilization cycles. Although ovarian hyperstimulation syndrome has been known for many years, it has been treated empirically and the underlying causes have been poorly understood. In order to reduce the risk of OHSS, several approaches have been proposed. One common technique is to coast the patient. In coasting, gonadotropin stimulation is withheld, and E2 levels are permitted to decrease before hCG is administered. Another technique, is elective cryopreservation of all embryos, plans to reduce the risk of OHSS by subsequent transfer of embryos in non-stimulated cycles. The present study compares the outcome of preventive strategies of ovarian hyperstimulation syndrome in ART cycles.

Since the inception of in vitro fertilization (IVF), the procedure has seen many advances that have significantly improved pregnancy rates as well as a reduction in complication rates.The benefits of blastocyst stage transfer have been established for the routine blind transfer technique of catheter introduction into the uterine cavity when a patient produces a large number of oocytes or has proven capacity to produce blastocysts. However, recent recommendations in using lower medication dosages for the controlled ovarian hyperstimulation, and in patients with lower response or advanced age, the number of developing embryos are limited and cleavage stage embryo transfers may be more advantageous. Also, since a significant number of embryos develop poorly beyond day three, early transfer is clinically prudent.32 consecutive patients with Infertility of various origins underwent Hysteroscopic Endometrial Embryo Delivery (HEED) on day two or three or day five after fertilization. Controlled ovarian hyperstimulation was done using standard protocols. Transvaginal oocyte retrieval was performed under local anesthesia with mild sedation. All women received some type of luteal support, be it progesterone or hCG. Oocytes were fertilized and cultured in early cleavage medium (Irvine Scientific) at 37 degrees C and 5% CO2 in air. Embryos were transferred at 48-120 hours post fertilization.The percent of total ongoing live births per transfer is not different between transfer on day two and three, and percent of ongoing pregnancies occurring with transfers on day five is higher than transfers on day three or two. (Chi square value for day two vs. five = 1.36, 0.5<p<0.1; chi square value for day three vs. five = 1.34, 0.5<p<0.1).Of pregnancies that occurred with transfer on day three, fifty percent resulted in miscarriage or biochemical pregnancy, while only sixteen percent had this result when transfer occurred on day two. Only one multiple pregnancy occurred with day of transfer two and five, while none of the transfers on day three produced a multiple pregnancy. Finally, no ectopic pregnancies were observed in either group.These results using Hysteroscopic Endometrial Embry Delivery (HEED) show for the first time that the technique is very effective with both cleavage stage embryos and blastocysts, including day two embryos. These results are even more remarkable due to our clinical bias to replace embryos on day 2 when presented with poor quality or low numbers of embryos. Our findings could offer new hope of success for a large number of poor prognosis IVF patients.

The role of inherited thrombophilias in RPL has generated a great deal of interest. This heterogeneous group of disorders results in increased venous or arterial thrombosis. Their associations with pregnancy loss rest on both proved and hypothetical alterations in placental growth and development, particularly placental vascular development. Abnormal placental vascularization and inappropriate placental thrombosis would link these thrombophilic states to pregnancy loss. Although some thrombophilic states may be acquired, most are heritable. Those heritable thrombophilias most often linked with reference to RPL include hyperhomocyteinemia. Activated protein C resistance associated with mutations in factor V, deficiencies in proteins C and S, mutations in the prothrombin gene promoter, mutations in prothrombin, and mutations in antithrombin III.Inherited thrombophilic mutations have been estimated to be causative in 50% of VTE during pregnancy. Approximately 40% of episodes of venous or arterial thromboembolic phenomena occur in patients carrying a heritable mutation. Associations between thrombophilias and adverse fetal outcomes cover a range of early gestational and obstetric disorders. These disorders include isolated and recurrent, early and late spontaneous pregnancy losses, intrauterine growth restriction (IUGR), intrauterine fetal demise (IUFD). Placental abruption and pregnancy- induced hypertension (PIH). This discussion will focus on pathophysiologic mechanisms, diagnostic tesing, and treatment strategies for patients with RPL who may have an inherited or acquired predisposition to thrombosis (excluding the antiphospholipid syndrome).The basis for the association between adverse fetal outcomes and heritable thrombophilias has focused on the mechanisms of impaired placental development and function secondary to venous or arterial thrombosis at the maternal-fetal interface.

PCOS is the commonest endocrinopathy in women and the commonest cause of unovutation in infertile couples.The role of endoscopy in the diagnosis and treatment of PCOS is well known. With laparoscopy we can see PCOS ovaries, differentiate other causes of infertility such as tubal problems or pelvic adhesion.With hysteroscopy we can see abnormal endometrial growth or polyps and we can do endometrial biopsy.Beside diagnostic aid, endoscopy can be used in the treatment of PCOS.In these cases we can do ovarian drilling with diathermy, Laser, crayon or hot water. And with this technic we induce ovutation, regulate the menstrual periods and increase the pregnancy rate.

Despite a growing collection of translation research and the development of new clinical protocols, laboratory and clinical outcomes in IVM remain suboptimal when compared to the standard IVF. One of the most popular approaches in improving IVM success is to supplement follicular development with gonadotropin stimulation (FSH and/or hCG) prior to oocyte collection. In this presentation, recent advances in the laboratory and embryological aspects of IVM cycles are described including oocyte identification and characterization, in vitro maturation, insemination, timing of embryo transfer and cryopreservation.In IVM cycles primed with hCG, oocyte identification is easier than in non- or FSH-primed IVM cycles due to the presence of an expanding cumulus mass around the oocytes. The immature oocytes with dispersed cumulus cells (CC) at collection in hCG-primed IVM cycles have high in vitro maturation and embryo developmental potential. Moreover, a few in vivo matured oocytes with dispersed CC can be obtained, and these have produced good quality embryos. Extending the period of hCG priming time before oocyte retrieval was one of the methods utilized to promote oocyte maturation in vivo and in vitro. The hCG could be given to patients when a dominant follicle (DF) reaches 10-12 mm to avoid detrimental effects on the sibling immature oocytes. ICSI has been suggested as the procedure of choice for inseminating mature oocytes and should be performed at least 1 h after the 1st polar body extrusion. Embryo transfer time depends on the quantity and the quality of the embryos produced after IVM. Vitrification is a more efficient method to freeze the embryos produced from IVM rather than slow freezing. In order to improve the IVM programs, it is essential to define not only the clinical aspects but also the laboratory and embryological aspects.

Introduction: The purpose of this review is to evaluate the management of implantation failure following ART cycles. The main limitation of the successful IVF-ET cycle is the low implantation rate of the in vitro embryos that reduce success rate from the more than 90% embryo transfer to less than 30% pregnancy per started cycles. It is suggested that unknown processes occur in the black box of the uterus and for this reason the most investigation in the art field are done in this part of the treatments. Embryo , endometrial receptivity and systemic condition are the most important factors in implantation. Nowadays many clinical trials have been done to improve the success rate and overcome of the implantation failure.Materials and Methods: Medline and Cochrane research.Results: Preimplantation genetic diagnosis is one of ways that it is very useful for the patients specially older women. Aneuploidiy of the gametes and embryos is very common and it related to the maternal age. Assisted zona hatching may be overcome zona hardening in older women. Uterine cavity abnormalities and endometrial polyp disturbe the implantation. So ,hysteroscopic evaluation of the endometrial cavity even without any pathological finding increases the implantation rate by stimulation of the endometrial gland secretion and lymphokines. In this process endometrial injury has positive effect on ART outcomes. Thrombophilia is associated with lower success and implantation rate. Although anti-thrombotic treatment such as low dose aspirin, heparin suppress the immunological activities but no significant benefits is shown in some of the clinical trials and it is controversial.Conclusion: PGD, correction of anatomical defect and increasing endometrial receptivity are the most important factors that can overcome on implantation failure and increase the success rate in ART cycles.

A major side-effect of controlled ovarian hyperstimulation (COH) in patients with polycystic ovary or polycystic ovarian syndrome (PCOS) is the risk of ovarian hyperstimulation syndrome (OHSS). In-vitro maturation (IVM) of immature oocytes represents a potential alternative for the fertility treatment of these patients. Recently, applications of FSH (hMG) or hCG priming have been used before immature oocyte retrieval to improve the success rate of IVM procedures.Although it is still controversial, hCG priming before oocyte retrieval seems beneficial in terms of easier oocyte retrieval, easier oocyte identification under stereomicroscope, maturation competence, and may increase the harvest of in vivo mature oocytes. Recently, new approaches attempted to improve the IVM program such as extending hCG stimulation or optimal hCG timing before immature oocyte retrieval. Therefore, as a first option, hCG-priming IVM treatment can be offered to women with PCO(S) instead of conventional IVF treatment with ovarian stimulation. In this session, I would like to introduce you to the recent progress of IVM cycles after hCG priming.

Endometriosis is defined as the presence of endometrial glandular and stromal tissue outside the uterus that induces a chronic inflammatory reaction. It may be present in up to 22% of asymptomatic women and 30% of women with unexplained subfertility.At present it is estimated that 10%-25% of all patients undergoing IVF are diagnosed with endometriosis, and 17%-44% of those also have ovarian endometriomas. In vitro fertilization has become the mainstay of treatment for endometriosis-related subfertility. Although there is some evidence to suggest that medical treatment with GnRH agonist can lead to a reduction in the size of the endometrioma by up to 51%, surgical removal of endometriomas remains the most effective approach for patients presenting with subfertility.Despite the lack of a firmly established causal relation between endometriosis and infertility, it is clear that treatment of endometriosis can improve fertility in some cases. Expectant management may be a reasonable approach in younger patients with early stage disease and a shorter duration of infertility. Current medical therapy is not efficacious, and its use should be discouraged as it may only serve to postpone conception. Laparoscopic surgery appears to be superior to expectant management or medical therapy in minimal-mild endometriosis and may also be of benefit for patients with advanced endometriosis. COH/IUI is a good option in mild and surgically corrected disease. In patients with earlystage endometriosis, IVF outcomes are similar to those with unexplained or tubal factor infertility, and Gn-RHa treatment combined with IVF may be useful for more advanced disease.In conclusion, the standard management of endometriosis in subfertile women before IVF remains controversial owing to the insufficient evidence to suggest superiority of one treatment strategy over another.All the therapeutic options, including conservative, medical, or surgical treatment, as well as the advantages and disadvantages should be fully discussed with the patient. Any decision for surgery should be carefully considered and balanced against the risks, especially in women with previous adnexal surgery or women with suboptimal ovarian reserve. If the woman opts for surgical treatment, she should be appropriately counseled about the potential risks of reduced ovarian function after surgery, including the remote possibility of oophorectomy.

Objective: To assess the efficacy of laparoscopic treatment on women with infertility.Methods: Retrospective analysis of 401 women with infertility underwent laparoscopic operation according to the laparoscopic findings.Results: The cumulative pregnancy rates were 50.0 % in the endometriosis group, 32.9 % in the tube obstruction group, 52.8 % in the polycystic ovarian syndrome (PCOS) group, 36.0 % in the uterine myoma group, and 42.0 % in the pelvic adhesion group respectively. 9310 %of women became pregnant within 18 months after operation.Conclusion: Laparoscopy is a simple, timely and effective method for the treatment of female infertility If the patient remains infertile in 12 ~ 18 months after operation, it is suggested that the patient should be treated with other assisted reproductive techniques.

Objectives: Uterine fibroids are one of the most common seen benign tumors of the uterus. The appearance of uterine fibroids has been linked to infertility in different degrees according to different locations, numbers, sizes and types. Considering surgery is the main treatment of uterine fibroids, the complications of surgery should be taking into account. Postoperative adhesion formation which may influence postoperative fertility. There is still no sufficient evident about myomectomy’s positive role in treating myomas. We analyze the different types, locations, numbers, sizes and many other factors related to myomectomy of fibroids’ different impact on postoperative fertility of patients with infertile complaints before surgery, to find out myomectomy’s role in treating infertility.Materials and Methods: 78 patients enrolled in the study, including 15 among them who were diagnosed as infertile Patients were follow-up for 2 years postoperatively. Datas of almost every aspect associated with uterine fibroids and myomectomy were collected, and analyzed.Results: With the use of multivariable logistic regression analyze, age has a P value of 0.02, with the OR value of 0.7 and the 95%CI (0.6, 1.0). This is the only independent factor which may influence the postoperative pregnancy rate, in a negative way. Other factors such as the presentation of infertility history, the location, size, number, entering the uterine cavity or not, types of the myoma and the surgery all have a P value of above 0.005 and the difference they have upon the postoperative pregnancy rate are not significant.Conclusions: To those who have uterine myomas, myomectomy may be an option for treatment, because postoperative pregnancy rate seems to increase significantly. Infertility may take place after the surgery for postoperative pelvic adhesion or potential factors that might cause problems on getting pregnant. For patients of whom uterine myomas seem to be the only problem, if have complaint of spontaneous abortion, problem may be solved after myomectomy. Age is the only independent factors that could influence fertility after surgery. The older the patient is the harder can she gets pregnant postoperatively. So early surgical treatment is recommended once myomas are found in the women in reproductive age. Myomas located in the anterior or posterior part of the uterine body have the similar impact on the postoperative pregnancy rate. Location, size, number, entering the uterine cavity or not, types of the myoma and the surgery do not affect the postoperative pregnancy rate.

Over millenia humans have shaped the genetic composition of today’s livestock. Traditional breeding schemes and, more recently, marker assisted selection strategies have been successfully used for incremental genetic improvement of livestock. In principle, transgenic technology, which creates the potential to enhance existing characteristics at unprecedented magnitude and speed, can be seen as the logical progression from these more traditional efforts to modify livestock genomes. Unlike traditional breeding and selection, the technology is not restricted by the species barrier and can utilise the gene pool of other species to introduce entirely novel and unique characteristics. The recent development of cell-mediated transgenesis for livestock based on somatic cell nuclear transfer allows for the introduction of a wide repertoire of genetic modification. This includes not only additive strategies to introduce a new gene function (gain of function) but also the deletion of gene functions (knockout, loss of function), replacing a gene function with a different one (knockin, exchange of function) or the transfer of genes in a spatial-temporal manner (conditional knockout). The use of the mammary gland’s high protein production capacity in dairy animals for the production of biopharmaceutical proteins in the milk has so far been the main driver for this technology platform due to strong economic incentive, ethical justification, greatest public acceptance and relative simplicity. The first biomedical product arising from a transgenic goat has recently been approved bringing transgenic technology into commercial reality. The genetic engineering of livestock provides also exciting opportunities to incorporate additional health benefits into important foods, such as milk, thereby creating entirely novel foods with unique properties not achievable by conventional means. Such food applications are however much more complex than biopharming and while presently in the research and development stage, their eventual introduction into the food chain remains some distance in the future.

Current transgenic livestock technology commonly relies on the random insertion of a gene construct into the genome. In this case, the activity of the transgene can be strongly influenced by the specific chromosomal context of the integration site, a phenomenon commonly referred to as position effect. As a consequence, the level of expression of a randomly inserted transgene can vary greatly or even result in adverse effects to the animals due to disruption of, or interference with, an endogenous gene. This unpredictability of transgene activity requires the generation and characterization of multiple transgenic cell and animal lines which is a major disadvantage for applications involving large animals due to the long gestation times and high costs involved. The randomness of the process can be avoided by the application of homologous recombination (HR) technology for the introduction of site-directed modifications. Due to the unavailability of livestock ES cells, HR approaches in livestock are presently restricted to the use of somatic cells which are hampered by very low efficiencies. The combination of the random chromosomal insertions of recognition sites for specific DNA recombinases (such as Cre or Flp) and the subsequent site-directed insertion of a transgene into this locus by a recombinase potentially offers much greater efficiencies. This recombinase mediated cassette exchange strategy provides a valuable system to efficiently produce transgenic livestock with predictable transgene expression levels. It also allows for the production of transgenic animals without antibiotic selection markers which are commonly used for the isolation of stably transfected cells but typically become part of the modified genome although they serve no useful function in the animal. An alternative approach is the use of episomal vectors which do not integrate into the genome but are maintained alongside the chromosomes as independent entities. They can thus ensure predictable expression and provide an elegant solution for potential problems associated with integration into the genome.However, emerging episomal vector systems are commonly of viral origin and dependent on viral sequences and factors. This has major drawbacks because the viral elements may be recognised as invading DNA and become permanently silenced or may even trigger the immune system, raising general biosafety concerns. The recent development of a novel self-replicating episomal vector system, based on the presence of a scaffold/matrix attachment region hold much promise as it can function independent of any viral factors. Moreover, the system has been engineered for the recombinase-mediated deletion of the bacterial vector backbone to limit the episomal vector that replicates in synchrony with the host cell chromosomes to its essential functional components. In the future, it will be important to demonstrate that these new concepts can live up to their promise and deliver greater predictability, efficiency and safety.

Objective: The first step for evaluation of male infertility is semen analysis but many studies suggest that other parameters such as sperm DNA integrity have more effects on ICSI outcomes. The aim of this study was to evaluate the correlation sperm DNA integrity using SCD and Comet assay with fertilization rate post ICSI.Materials and Methods: DNA damage in sperm of 41 men undergoing ICSI treatment was measured by comet assay and SCD test. Around 16–18 h post ICSI, fertilization was assessed by presence of pronuclei and the percentage of fertilization was calculated by the ratio of fertilized oocytes to the total number of survived injected metaphase II (MII) oocytes multiplied by 100. Coefficients of correlation were calculated using SPSS 11.5 software.Results: No significant correlation was obtained between fertilization rate and sperm DNA damage measured by SCD (p=0.091), and Comet assay (p=0.612) in ICSI patients.Conclusion: Our results show that the sperm DNA damage doesn't directly effect on fertilization rate, since damaged DNA carried into the zygote by the fertilizing spermatozoon will be repaired by the oocyte. However, it can be effects on subsequent embryo development. It could be devised to identify and select spermatozoa with intact DNA or to remove spermatozoa with damaged DNA so as to improve ICSI outcomes.

Objective: Alcoholism in men causes increase in abnormal shaped sperms that can lead to impotency. Abnormal liver function and high estrogen levels may cause interference with sperm development which can severely suppress sperm. Moreover, alcohol abuse has been linked with damaged sperm and reduced sperm counts. . Like a toxin, alcohol may destroy the sperm-generating cells. In addition to this, alcohol abuse can also have adverse effects on the hormone levels in men. Which may interfere with sperm development and hormone levels? Alcohol is also a toxin that can kill off the sperm-generating cells in the testicle. Worse still, chronic alcoholism can cause atrophy in the testicles, gynecomastia and the lack of sexual interest. Excessive alcohol consumption causes a disorder in the gonads resulting in changes in the structure of the testicles and decline in the T serum level. A drop in the T serum level can cause male infertility. In addition, alcohol can result in abnormalities in sperm size, shape and the sperm tail. Sperm motility can also be affected negatively. This change in sperm shape can seriously compromise the viability of the spermatozoa. Also, chronic alcoholism damages the sperm morphology that often has irreversible effects.Materials and Methods: 42 adult male Wistar rats were randomly divided into 6 groups (n=7 each) with two groups serving as control, in the treatment groups were received 10% ethanol in distilled water for 35,70 and 120 days. All rats were sacrificed by CO2 inhalation and testis tissues were removed and prepared for Histological (H & E, Vaigrate Iodine and Toloedine blue (for mast cells) staining) and Histochemical (by cryosection and Lipase, Oil Red O staining methods) study. In addition spermatozoa were removed from cauda epididymis and analyzed for sperm motility, concentration in the cauda epididymis, viability and sperm chromatin quality and DNA integrity was assessed by Aniline blue and Acridine Orange staining following sperm sample preparation. Serum testosterone and estrogen level was determined by radioimmunoassay technique.Results: This study confirmed that treated by Ethanol had significant decrease the testosterone level in plasma. Ethanol had significant increase the DNA Damage and chromatin abnormality in the cauda epididymal spermatozoa as evidenced by Acridine Orange (AO) and Aniline blue staining respectively. Treatment of male rat with Ethanol caused significant decrease in sperm count, motility, and viability, while abnormal sperms increased as compared to control. Histological and histomorphometrical study confirmed that Ethanol had significant decrease the semniferous tubules diameter, number of germinal cells, and spermatogenesis in the semniferous tubules and increase the connective tissue and appearance of vacuolated edema in the interstitial connective tissue between the seminiferous tubules and with increased hypotrophic mass of leydig cells. Toloidine blue staining confirmed that Ethanol had significant increase the number of Mast cells in testis and epididymis tissues. Histochemical study by Oil Red O and Lipase methods confirmed that Ethanol had significant increase the fat droplets in the semniferous tubules and lipase reaction confirmed that the Ethanol had increase the lipase enzyme in the testis tissue.Conclusion: Conclusion Yes, drinking alcohol can adversely affect on fertility and also cause damage to baby. The male reproductive system consists of the hypothalamus, the anterior pituitary gland, and the testes. Alcohol can interfere with the function of each of these components, thereby causing impotence, infertility, and reduced male secondary sexual characteristics. In the testes, alcohol can adversely affect the Leydig cells, which produce and secrete the hormone testosterone. Studies found that heavy alcohol consumption results in reduced testosterone levels in the blood. Alcohol also impairs the function of the testicular Sertoli cells that play an important role in sperm maturation. In the pituitary gland, alcohol can decrease the production, release, and/or activity of two hormones with critical reproductive functions, LH and FSH. Finally, alcohol can interfere with hormone production in the hypothalamus.

Objective: PATZ1 is a recently discovered zinc finger protein that, due to the presence of the POZ domain, acts as a transcriptional repressor affecting the basal activity of different promoters. To gain insights into its biological role, we generated mice lacking the PATZ1 gene. Male PATZ1-/- mice were unfertile, suggesting a crucial role of this gene in spermatogenesis. Consistently, most of adult testes from these mice showed only few spermatocytes, associated with increased apoptosis, and complete absence of spermatids and spermatozoa, with the subsequent loss of tubular structure.Materials and Methods: The analysis of PATZ1 expression, by Northern blot, Western blot and immunohistochemistry, revealed its presence in Sertoli cells and, among the germ cells, exclusively in the spermatogonia.Results: Since PATZ1 has been indicated as a potential tumour suppressor gene, we also looked at its expression in tumours deriving from testicular germ cells (TGCTs). Although expression of PATZ1 protein was increased in these tumours, it was delocalized in the cytoplasm, suggesting an impaired function.Conclusion: These results indicate that PATZ1 plays a crucial role in normal male gametogenesis and that its up-regulation and mis-localization could be associated to the development of TGCTs.

Objective: There is lots of oxygen reaction element in cigarette smoking that induce reactive oxygen stress (ROS) in the body ROS can make DNA damage in somatic and germ cells. The purpose of this study was to evaluate the effect of smoking on sperm DNA integrity in infertile patients by comet assay.Materials and Methods: DNA damage in sperm of 72 men undergoing IVF and ICSI treatment was measured by comet assay and DNA damage was compared between smoker and nonsmoker groups by t-test using SPSS 11.5 software.Results: The result show that significant different between sperm DNA integrity among smoker and non smoker groups (p=0.05).Conclusion: it can be concluded that the cigarette prone sperm DNA damage.

Objective: To evaluate the efficacy and safety of most popular selective serotonin reuptake inhibitors (SSRIs) drug (citalopram, escitalopram, dapoxetine, paroxetine, venlafaxine), and tramadol and pindolol in delaying ejaculation in patients with premature ejaculation (PE).Materials and Methods: A predetermined number of married men with PE were randomly assigned to receive the study drug or placebo. Pretreatment evaluation included history and physical examination, intravaginal ejaculatory latency time (IELT), International Index of Erectile Function (IIEF) and Meares-Stamey test. The efficacy of each treatment was assessed every 2 weeks during treatment, at the end of study using responses to IIEF, IELT evaluation, mean intercourse satisfaction domain, mean weekly coitus episodes and adverse drug effects.Results: The IELT after citalopram and placebo gradually increased from 32 and 28 seconds to approximately 268 and 38 seconds, respectively. The mean IELT after dapoxetine and placebo increased from 28 and 31 seconds to approximately 193 and 54 seconds, respectively (P =0.001). At the end of trial with dapoxetine, paroxetine, and placebo the mean IELT was increased from 38, 31 and 34 seconds to 179, 370 and 55 seconds, respectively. The mean IELT increased from 31 and 29 seconds to 516 and 54 seconds with escitalopram and placebo, respectively (P=0.001). The geometric mean IELT in paroxetine-pindolol and paroxetine-placebo group demonstrated 3.7 (95% confidence interval (CI): 2.16-5.26) and 1.7 (95% CI: 0.82-1.81) fold-increase, respectively (P=0.001). The mean IELT after tramadol and placebo increased from 19 and 21 seconds to approximately 243 and 34 seconds, respectively ( P <0.001). The geometric mean IELT in venlafaxine and placebo group demonstrated 1.7 (95% CI; 0.76-1.96) and 1.6 (95% CI: 0.87-1.84) fold-increase, respectively (P=0.1).Conclusion: Oral citalopram and escitalopram is a highly effective treatment for PE with long-term benefit for the patient after it is withdrawn. Paroxetine appears to provide significantly better results in terms of IELT and intercourse satisfaction versus dapoxetine, and tramadol. Venlafaxine is no better than placebo in treatment of PE. A single high dose of pindolol (7.5 mg) is an effective augmentation strategy in paroxetine-refractory patients.

Objective: Spermatogonial stem cells (SSCs) are unique population of adult stem cells in mammalian testes which continuously provide gametes and transfer genetic material to the next generation. Various feeder layers have been tested to support the in vitro culture of SSCs; however, age effect of feeder cells has been remained a controversial issue. This study was initiated to compare Sertoli cells derived from neonatal and adult mice to examine age effect of Sertoli cells in the maintenance and proliferation of mouse SSCs in vitro.Materials and Methods: SSCs were isolated from testes of 6 day-old mice and culture in vitro at the presence of Glial-derived neurotrophic factor (GDNF) for 10 days and then transferred to Sertoli cells isolated by DSA lectin from neonatal (6 day-old) and adult (6-8 week-old) mice. After 5 days, area and number of SSC colonies were measured. Immunostaining was used to detect expression of spermatogonial markers including a6/b1-Integrin, C-kit and Oct-4. In addition, SSC colonies were harvested at day 5 and the percentage of a6/b1-Integrin-positive cells was measured by flowcytometery. Transplantation assay was used to confirm the stemness of spermatogonial cells.Results: Immunostaining analysis showed that our culture system contained SSC colonies as they were positive for a6-Integrin, b1-Integrin and Oct-4 and negative for C-kit. In addition, these stem cells were functional as they were able to migrate to seminiferous basal membrane after transplantation to testes of busulfan-induced infertile adult mice, Results showed 5 days after co-culture of SSCs with Sertoli cells, the area and number of colonies and number of cells in each colon were significantly higher on Immature Sertoli cells. Moreover, results showed that colony efficiency of cultured SSCs on Immature Sertoli cells was significantly higher than other two groups. Flowcytometry analysis revealed that there was significant increase in the number of a6-Integrin-positive cells and b1-Integrin-positive cells in the culture with Immature Sertoli cells (72.9±14.9 %)(67.4±7.7) in contrast to culture with Mature Sertoli cells (29.5±6.5 %)(52.9± 1.9).Conclusion: In conclusion, the number of SSCs was higher in co-culture with Immature Sertoli cells. It could be referred to difference in microenvironments that Immature and Mature Sertoli cells provide for in vitro culture of SSCs. As SSCs was derived from neonatal mice, another interpretation is that more suitable culture condition could be obtained when SSCs and feeder Sertoli cells are derived from mice in the same age; either from neonatal mice or from adult ones. Further studies would confirm these hypotheses.

Objective: Varicocele occurs in approximately 15% to 20% of the general male population and it is the most common cause of poor semen production and decreased semen quality. It has been demonstrated that patients with varicocele have a significantly higher DNA fragmentation index (DFI) and spermatozoa with nuclear anomalies than healthy fertile men. Therefore, the aim of this study was to evaluate sperm chromatin integrity in these patients.Materials and Methods: Sixty men referring to the andrology laboratory were categorised into three different groups: 20 infertile men with varicocele, 20 infertile men with abnormal semen parameters and 20 fertile men who had normal spermatogram were considered as control group. Semen analysis was performed according to WHO criteria. To evaluate sperm chromatin quality and DNA integrity, after fixation of sperm smears, aniline blue, toluidine blue, chromomycin A3 and acridine orange staining were applied in three groups. The slides were analysed by light and fluorescent microscopy and to determine the percentage of mature or immature spermatozoa, 200 spermatozoa were counted in each slide.Results: The results showed that the rates of aniline blue-reacted spermatozoa were significantly higher in infertile and varicocele patients than in the normal group (p < 0.001). In addition, with regard to chromomycin A3, acridine orange and toluidine blue staining, there was a significant difference between the three groups (p < 0.001). The results showed that the varicocele samples contain a higher proportion of spermatozoa with abnormal DNA and immature chromatin than those from fertile men as well as infertile men without varicocele.Conclusion: Therefore, varicocele results in the production of spermatozoa with less condensed chromatin and this is one of the possible causes of infertility due to varicocele.

Objective: To compare specific nutrient intake between normospermic and oligoasthenoteratospermic patients attending infertility clinics in two Mediterranean provinces of Spain.Materials and Methods: Case-control study. Setting: Private fertility clinics in southeastern Spain. Patient(s): Thirty men with poor semen quality (case subjects) and 31 normospermic control subjects of couples attending our fertility clinics. Intervention(s): We recorded dietary habits and nutrient consumption using a food frequency questionnaire adapted to meet specific study objectives. Main Outcome Measure(s): We calculated nutrient intakes by multiplying the frequency of use for each food by the nutrient composition of the portion size specified on the food frequency questionnaire and by addition across all foods to obtain a total nutrient intake for each individual. Semen quality was assessed by measuring volume, concentration, motility, and morphology. Hormones levels were also analyzed in case and control subjects.Results: In the logistic regression, control subjects had a significantly higher intake of carbohydrates, fiber, folate, vitamin C, and lycopene and lower intakes of proteins and total fat.Conclusion: A low intake of antioxidant nutrients was associated with a poor semen quality in this case-control study of Spanish men attending infertility clinics

Objective: Controlled ovarian stimulation (COS) has been shown to advance endometrial maturation and affect adversely implantation in assisted reproduction technology (ART) cycles. It has been reported that there is a better embryo-endometrium synchrony in frozen-thawed embryo transfer (FET) cycles than fresh embryo transfer (ET) cycles. The objective of this study was to compare ongoing pregnancy rate between fresh ET and FET cycles in ART.Materials and Methods: In a prospective, controlled study, the patients who were classified as high responders were randomized to either fresh ET or FET. The embryos in FET group were cryopreserved with vitrification by Cryotop method. Randomization was done on the day of ET according to a computer-generated random numbers. Ongoing pregnancy rate was the primary outcome measure.Results: A total of 374 patients were included, 187 of which were randomized to FET and 187 to fresh ET. There were 39% (n=73) ongoing pregnancy in FET group compared with 27.8% (n=52) in fresh ET group [odds ratio (OR) = 1.66; 95% confidence interval (CI) = 1.07-2.56; p < 0.05]. Implantation, clinical pregnancy and multiple pregnancy rates were also higher in FET group.Conclusion: FETs can be performed instead of fresh ETs to improve the outcome of ART cycles in highly selected patients.

Objective: The purpose of this study was to evaluate the effect of fibroblastic growth factor on resumption of meiosis, in vitro maturation of immature mouse oocytes and resulting embryo development with and without FGF.Materials and Methods: Cumulus - oocyte complex (COCs) and germinal vasicle (GV) were obtained from female NMRI mice 46-48 hours after administration of an i.p. injection of 5 IU PMSG. COCs were released from larg antral follicles and culture for 18 hours in humidified atmosphere with 5% CO2 at 37oC in TCM199 supplemented with 0, 10, 20, 50 and 100ng/ml FGF. After in vitro maturation (IVM), metaphase ІІ (MІІ) oocytes were co-incubated with sperms for 4 hours in T6 medium. For all groups, 2PN embryos were cultured in the same medium and cleaved embryos was assessed after 48 hours.Results: FGF increased the proportion of in vitro growing (IVG) oocytes reached metaphase ІІ in all compared rate, recorded in the 20 ng/ml FGF treatment groups (42.8%), was significantly higher (p<0.05) than the 10 ng/ml (31.1%), 50 ng/ml (9.5%) and 100 ng/ml FGF (6.6%) containing treatment groups but no significant difference was found as compared to control (29%).Conclusion: Exogenous FGF during IVM improved the nuclear maturation and embryo development.

Objective: Reprogramming of nuclei allows the dedifferentiation of differentiated cells. Somatic cells can undergo epigenetic modifications and reprogramming through their fusion with embryonic stem cells (ESCs) or after overexpression of a specific blend of ESC transcription factor-encoding genes. Our goal was to demonstrate that the activation of Wnt/beta-catenin signaling triggered somatic cell reprogramming.Materials and Methods: We performed cell fusion experiments by fusing ES cells with neural stem cells or ES cells with Thymocytes or ES cells with Mouse Embryonic Fibroblasts. ES cells or the hybrids were treated with Wnt3a or BIO (Wnt pathway activator) or Dkk1 (Wnt pathway inhibitor). Fusion between ES cells over-expressing different amounts of beta-catenin with NS cells were also carried out. Reprogrammed cells were analyzed in vivo and in vitro for their ability to differentiate in different cell types. Expression of stem cell markers and methylation profile of stem cell promoters was also analyzed in the reprogrammed cells. For further details see Lluis et al. Cell Stem Cell 2008.Results: We showed that cyclic activation of Wnt/beta-catenin signaling in ESCs with Wnt3a or the glycogen synthase kinase-3 (GSK-3) inhibitor 6-bromoindirubin-3’-oxime (BIO) strikingly enhanced the ability of ESCs to reprogram somatic cells after fusion. In addition, we showed that reprogramming is triggered by a dose-dependent accumulation of active beta-catenin. Reprogrammed clones expressed ESC-specific genes, silenced somatic differentiation markers, became demethylated on Oct4 and Nanog CpG islands, and were able to differentiate into cardiomyocytes in vitro and to generate teratomas in vivo.Conclusion: Our data thus demonstrate that in ESCs, periodic beta-catenin accumulation via the Wnt/beta-catenin pathway provides a specific threshold that leads to the reprogramming of somatic cells after fusion.

Objective: Amniocentesis is the most common test used for prenatal diagnosis of a chromosome problem in the fetus. It involves the removing a small amount of amniotic fluid which surrounds the fetus in the amniotic sac. The amniocentesis sample is sent to the laboratory where the cells are cultured and then the chromosomes are analyzed. A new supplemented medium has been developed to improve amniotic fluid cell growth in mouse.Materials and Methods: The medium consists of a mixture of Ham's F12 medium and Dulbecco's modified Eagle's medium (DMEM) supplemented with antibiotics, and 20% fetal calf serum (FCS).Results: Good clonal growth is achieved consistently in 4-6 days without any mycoplasma contamination. The volume of amniotic fluid required to initiate culture can be as little as 1 ml. Amniotic fluid samples contaminated with red blood cells with no visible clot also grow well.Conclusion: The results of amniocentesis will indicate the likelihood of the fetus developing certain chromosomal conditions, such as Down's syndrome, Edward's syndrome, and Patau's syndrome, which are all conditions where the fetus is born with an extra chromosome.

Objective: The aim of this study was to evaluate the effect of vitrification with two methods of 4 and 2-step dehydration on sheep ovarian tissue.Materials and Methods: Sheep ovarian tissue samples were collected and allocated to three groups of fresh, 4-step vitrified and 2-step vitrified. A modified carrier and vitrification solution was applied. The proportion of morphologically intact follicles in fresh ovarian tissues was compared with that in vitrified-warmed tissues. The base medium (BM) containing hepessed tissue culture medium (HTCM) supplemented with 10% human serum albumin (HSA) was used as the solvents for vitrification and warming solutions. The ovarian tissue pieces were dehydrated by using regimens; 4-step: (1) 3.75% ethylene glycol (EG) and dimethyl sulfoxide (DMSO); (2) 7.5 % EG and DMSO; (3) 10% EG and DMSO; (4) 15% EG and DMSO + 0.25 mM sucrose in the base medium each for 5 minutes at 4°C and 2-step: (1) 7.5% EG and DMSO; (2) 15% EG and DMSO + 0.25 mM sucrose in the base medium each for 10 minutes at 4oC. Also imaging of apoptotic follicles was performed in fresh and vitrified tissues using TUNEL protocol.Results: The proportion of intact antral follicles in the fresh (66.66%±0.33) and 2-step vitrified (56.66%±0.16) groups were significantly higher than that in the 4-step vitrified (0%) group whereas the difference of this proportion between fresh and 2-step vitrified was not significant But between the three groups in the proportion of primordial, primary and preantral follicles the differences was not statistically significant. The apoptosis imaging of ovarian pieces also did not show statistically differences between all the groups.Conclusion: These results indicated that sheep ovarian tissue vitrification by 2-step method is simpler and more effective than those of 4-step method. On the other hand, as sheep and human ovarian tissue are more similar, this technique can be exam for cryopreservation of the human ovarian tissue too.

Objective: This study was conducted to compare the expression of maturation genes of sheep cumulus-oocyte complexes follow vitrification by conventional and cryotop methods.Materials and Methods: cumulus-oocyte complexes (COCs) were harvested from slaughtered sheep ovaries. COCs were divided into three groups: Control, Conventional Vitrification & Cryotop Vitrification. In control group, COCs were transferred immediately to in vitro maturation medium (IVM). Vitrification was done by DMSO & EG. The viability of vitrified-warmed COCs was assessed morphologically. Vitrified-warmed COCs were matured like the control group and oocytes nuclear stage was determined by Hoechst staining. Oocytes were subjected for assessment of maturation genes expression by Real-Time quantitative RT-PCR.Results: Cryotop vitrification had higher percent of healthy COCs after warming (83.84%) and also showed a significant difference with conventional vitrification. Mature oocytes (MII) were 51.94% and 48.81% in the control and cryotop groups respectively. Conventional vitrification showed a significant difference with control group in the expression of GDF9, BMP15 & BMPRII genes, also GDF9 & BMP15 in the conventional vitrification had a significant difference with cryotop vitrification. The relative expression of BMP15 & BMPRII was significantly different between control and cryotop groups. ALK5 expression was evaluated too.Conclusion: According to the less success of immature sheep oocytes cryopreservation, it seems that vitrification by cryotop can reduce cryoinjuries and increase the viability, post-thaw quality and maturation rate of COCs. This kind of vitrification causes the least expression changes of maturation genes in comparison with conventional vitrification.

Objective: This study investigates the number and size of ovarian antral follicles in relation to plasma follicle stimulating hormones (FSH) and luteinizing hormone (LH) concentrations from birth to 26 weeks of age in ewe lambs of the Ouled Djellel breed, a non-seasonal breed of sheep.Materials and Methods: Plasma was collected from 10 ewe lambs at 14 sampling times (Week 0, i.e. <24 h, Week 1 and every two weeks from Week 4 to Week 26, inclusive). At each of these stages, four ewe lambs were slaughtered, the ovaries recovered and weighed, and the number and size of the follicles determined from histological examination.Results: The pattern for plasma FSH showed a peak at Week 10, a smaller peak at Week 18 and a very small peak at Week 24. The pattern for LH was similar until Week 24 when the largest peak occurred. Paired ovarian weight increased rapidly from birth to four weeks and then more slowly to 10 weeks, followed by a decline at 12 weeks and a gradual increase from 14 to 24 weeks of age. The number and total diameter of follicles ³3 mm in diameter showed similar patterns of development - rising gradually from birth to week 14, falling to week 16 and then rising more rapidly to a peak at week 24. Maximum follicle diameter declined from birth to Week 1, then rose rapidly to Week 4, followed by a more gradual rise to week 14 and, thereafter, a more rapid increase to a peak of 7.23±0.16mm at 24 weeks old. The number of follicles (<3mm diameter) increased rapidly from birth to week 10 and then declined to values similar to those at weeks 1 and 4. First behavioural oestrus was observed at week 24 and a corpus luteum was present on the ovary of one lamb at Week 24 and two lambs at week 26.Conclusion: It was concluded that two or three peaks in plasma FSH and LH levels precede puberty and first and the increase in follicle numbers and size generally reflected the pattern of plasma FSH and LH levels.

Objective: Oocyte maturation is a process that during which the oocyte attains the competence to be fertilized. For clinical application, most oocytes are obtained from ovarian follicles of women who have been treated with hormones. Side effects of superovulation are ovarian hyperstimulation syndrome or polycystic ovary syndrome. Hence, there is an increasing interest in retrieving oocytes without gonadotropins and then in vitro maturation. Hydrostatic pressure as a physical force is effective on reproductive system. Obviously, there is an increase in intrafollicular pressure 15-20 mmHg in the ovulating follicle during the late stage of the ovulatory process. In this study, we examined the effect of hydrostatic pressure on in vitro maturation of oocytes derived from the stimulated and unstimulated ovary.Materials and Methods: In this study we had two experimental groups. In experiment I, female NMRI mice 6-8-week-old received 10 IU/ml PMSG hormone and in experiment II did not. Preovulatory follicles were isolated from mice ovaries, each follicle cultured individually in microdrops of MEM-α supplemented with 5% fetal bovine serum, 100-mIU/ml rFSH (Gonal-f), 10ng/ml recombinant Epidermal Growth Factor, 7.5 mIU/ml HCG under mineral oil for maturation. At the start of in vitro maturation follicles from two experiments divided into two treatments. In treatment I follicles were transferred to pressure chamber and exposed to 20 mmHg hydrostatic pressures for 30 min and in treatment II follicles were transferred to pressure chamber without hydrostatic pressure exposure. Then follicles were cultured for maturation of oocyte.Results: After 24 h in experiment I percent of MII oocyte was increased in follicles with hydrostatic pressure exposure (15.38%) compared to follicles without hydrostatic pressure exposure (10%) (P<0.05). In experiment II percent of MII oocyte was increased in follicles with hydrostatic pressure exposure (18.42%) compared to follicles without hydrostatic pressure exposure (10.52%)(P<0.05). After 48 h in experiment I, percent of MII oocyte was increased in follicles with hydrostatic pressure exposure (38.46%) compared to follicles without hydrostatic pressure exposure (20%)(P<0.05). In experiment II percent of MII oocyte was same in follicles with hydrostatic pressure exposure (44.8%) and in follicles without hydrostatic pressure exposure (42.1%).Conclusion: Gonadotropin stimulation is used to superovulation, achieves multifollicular recruitment, but reduces oocyte in vitro maturation. According to our findings hydrostatic pressure can be improve oocyte in vitro maturation by positive effect on the preovulatory follicle.

Objective: LIF is a 45-56 kDa glycoprotein that has important role in proliferation and embryo implantation. LIF affects its function via its receptors, such as gp130. Effects of LIF in oocyte matuiration and expression of its receptor in oocytes is unknown.Materials and Methods: Immature mice superovulated with HMG and GV oocytes obtaind from ovary 48 hours after. The GV oocytes were cultured in TCM199 with 5% Co2 for obtaining MII oocyte. For Real-time RT-PCR, Total RNA from GV and MII oocytes were extracted by Trizol reagent the quantity and quality of RNA were determined by spectrophotometry and electrophoresis, respectively. Reverse transcription was performed by Super Script III reverse transcriptase with 1 mg of total RNA followed by DNaseI treatment and heat inactivation. Expression of gp130 was determined by Real time RT-PCR.Results: Our results showed that gp130 is expressed neither in GV nor in MII oocytes during in vitro maturation of mouse oocytes. It is proven that gp130 is expressed in human oocyte but is not expressed in mouse oocytes.Conclusion: It is suggested that in mouse, LIF could affects the oocyte via another receptor. However its details must be elucidated.

Objective: we compared ROS levels in seminal plasma of infertile men with this level in healthy donors. We also determined the ROS level association with percentages of different sperm morphological abnormalities, the sperm deformity index, and the teratozoospermic index scores.Materials and Methods: In total 30 infertile patients and 25 healthy donors as control were selected. Sperm analysis was done for all patients. Azoospermic patients were excluded from the study. ROS level in semen were measured by a chemiluminescence assay in both group. Thin smears af well-mixed semen were stained with papanicolaou to assess morphological abnormalities.Results: The mean ROS level in normal men was 166.10 RLU (Relative Light Unit) while this was 1630.50 RLU in infertile patient which is significantly higher in case group (p = 0.000). A significant positive correlation was observed between sperm ROS production and the proportion of sperm with abnormal morphology. ROS production was positively correlated with the proportion of sperm with amorphous head, midpiece defects, cytoplasmic droplets, tail defect, pinhead sperms, SDI scores, and TZI scores (p<0.005).Conclusion: The level of ROS in seminal fluid of infertile men is significantly higher than fertile donors and also sperm morphological abnormalities, SDI score and TZI score are useful to predict levels of ROS.

Objective: Positive effects of sperm and seminal plasma on female genital tract, embryo vitality and implantation had been explained in some animal and human experiments. But there is a doubt to advise intercourse or abstinence in infertile couples during the in-vitro fertilization (IVF) cycles. We designed a prospective controlled trial to find out the intercourse effect on intra cytoplasmic sperm injection (ICSI) cycle outcome among of 1321 fresh embryo transfer. Here we report the results of pregnancy and implantation rate in 225 cycles of carrying 496 embryos which had been included till now.Materials and Methods: A total of 225 patients undergoing IVF/ICSI treatment between October and December of 2008 were randomly allocated into the case (107 people) and the control group (105 people). It was explained in case group to have intercourse 12 to 24 hours before ovum pickup and 12 hours after embryo transfer. Control group were forbidden to have intercourse in 72 hours before until 2 weeks after embryo transfer. All embryos were cultured and transferred in the same procedures. Chemical pregnancy was established in 14 and 17days after transfer by measuring bHCG level in plasma and clinical pregnancy in 6 weeks after transfer by appearance of gestational sac in ultrasonography.Results: Among 225 patients who were participated in trial, 13 cycles resulted to no embryo transfer (5.8%). Mean ages of couples, duration and cause of infertility, number of oocyte, fertilization rate, source of injected sperm and sperm parameters in each group were the same.496 embryos in 212 people were transferred totally which resulted in 64 pregnancies (30.1%). Pregnancy rate in case group was 31.8% versus 28.6% in control group; the difference was not statistically significant. Both bHCG level differences were significant between groups (p<0.05). Number of gestational sac was the same in both groups. Conclusion: This study supports the findings of other similar experiments which indicate that semen could enhance the endometrium and promoting the pregnancy. Although the pregnancy rate was the same in this preliminary analysis but the higher bHCG level in case group was significant. It suggests that intercourse may probably make a better chance for pregnancy but of course it is necessary to study a large number of patients. At this time we can suggest that there is not necessary to advise all patients to have abstinence during IVF cycle.

Objective: Here in, we investigated the embryonic-abembryonic axis of the blastocyst in the porcine species. To avoid the influences of the fertilization cone, which indicates the sperm entry position, and to prevent topological change of the two or more apposing pronuclei in the egg center caused by polyspermy after IVF, we chose porcine parthenogenetic embryos for use in the present study even though they may not represent normal embryonic development in the pig. Here in, we describe the fate of an individual blastomere from a 2-cell-stage parthenogenetic porcine embryo.Materials and Methods: For lineage tracing, DiI, a fluorescence dye, was injected into only a blastomere of the 2-cell stage parthenogenetic porcine embryos. If the first blastomere to divide was labeled, the embryo was included in the leading group, and while all others were included in the lagging group. Mitochondrial distribution of 2-cell parthenotes was also examined by using MitoTracker Green staining and confocal system.Results: In 60.5% of the blastocysts in the lagging group, the progeny of the labeled blastomeres formed the inner cell mass (ICM) and adjacent trophectoderm (TE) hemisphere; 62.1% of the blastocysts in the leading group had progeny of the labeled blastomeres distributed only to the TE (opposite of ICM). The rest of the lagging and leading groups showed random distributions. Unlike murine parthenotes, biased mitochondrial distribution was also found in porcine parthenotes (38.1%).Conclusion: Our findings indicate that the ‘leading’ blastomere of the 2-cell porcine parthenote forms the distal TE (abembryonic) and that the ‘lagging’ blastomere forms the remaining portion of the blastocyst, including the ICM (embryonic). Biased distribution of mitochondria in each 2-cell blastomere may contribute partly to this event. This study was supported by Korea Science and Engineering Foundation (KOSEF) grants funded by the Government of the Republic of Korea (MOST; M10641000001-06N4100-00110 and R01-2007-000-10316-0).

Objective: a) To identify the endometrial factors which are directly or indirectly regulated by progesterone in nonconception cycle and also to investigate whether their expression is modulated during early pregnancy in primates and b) To identify the factors which are differentially expressed in endometrial tissues and uterine secretions during the progesterone dominant or mid-secretory phase as compared to the estrogen dominant or proliferative phase in humans.Materials and Methods: For objective a, six regularly cycling healthy female bonnet monkeys (Macaca radiata) were subcutaneously injected with Onapristone- ZK 98.299, an antiprogestin, dissolved in vehicle (benzyl benzoate: castor oil, 9:1), at a dose of 5.0 mg starting from day 1 of the menstrual cycle and continued every third day for one cycle. Onapristone treatment rendered the animals infertile or implantation incompetent due to induction of endometrial nonreceptivity. Control animals (n=6) were treated with vehicle alone. Circulatory steroid levels were estimated in animals before and after the treatment using radioimmunoassays (Sachdeva et al, 2001; Patil et al, 2005). Endometrial biopsies were collected from both onapristone-treated and vehicle-treated animals on day 8 estradiol peak. Immunohistochemistry and reverse transcriptase polymerase chain reaction were used to investigate whether some of the select factors were differentially expressed after the blockade of optimal progesterone action on endometrium in bonnet monkeys. Differential display reverse transcriptase polymerase chain reaction (DDRTPCR) and 2D proteomics approaches were also used to identify the factors which were differentially expressed in the endometrium of implantation incompetent bonnet monkeys, compared to vehicle treated control animals. To investigate whether the expression of some of these factors is altered during pregnancy, endometrial samples were collected from another group of animals on day 6 of pregnancy (approximately equivalent to day 8 post estradiol peak). Towards this, regularly cycling female bonnet monkeys (n=6) with normal hormonal profiles (peak estradiol levels- 300-600 pg/ml; progesterone levels (3-6 ng/ml) were mated with males of proven fertility for six continuous days starting from two days prior to the expected estradiol peak. Pre-implantation factor (PIF) in the sera was used as a surrogate marker of pregnancy (Rosario et al, 2005a). The control group included nine PIF negative animals. For objective b, regularly cycling women (21-35 years) of proven fertility with a history of at least one live birth were enrolled in the study. Ovulation was monitored by serial ultrasonography (USG) to ascertain the follicular collapse. Endometrial tissue and uterine fluid samples were collected from women on day 6 post-ovulation (in mid-secretory phase) or on day 2-3 prior to ovulation (in proliferative phase). 2D proteomics and immunoblot analysis were used to identify the factors, which were differentially expressed during the progesterone dominant phase as compared to the proliferative phase in human endometrial tissues and uterine fluid samples.Results: Candidate factor approach revealed differential expression of several cytokines such as interleukin 1 beta, interleukin 6, transforming growth factor beta, leukemia inhibitory factor (Sachdeva et al, 2001) and cell adhesion molecules like alpha v and beta 3 integrin (Puri et al, 2000) in the mid-secretory phase endometria of antiprogestin treated bonnet monkeys as compared to that of control animals. This suggested that progesterone directly or indirectly regulates the expression of these factors. Interestingly, expression of interleukin 6, transforming growth factor beta in endometrium was significantly higher in the endometria of pregnant animals, as compared to that in nonpregnant animals (Rosario et al, 2005b), whereas the expression of leukemia inhibitory factor did not alter significantly during early stages of pregnancy. Integrins alpha v and beta 3 also showed cell type specific increase in endometrium during early stages of pregnancy (Nimbkar-Joshi et al, unpublished). These studies indicated that progesterone priming during nonconception cycle leads to increase in the expressions of endometrial TGF beta, LIF, interleukin 6, integrins and these factors probably facilitate endometrial preparation for implantation. Expressions of some of these factors are further modulated during early stages of pregnancy. Functional genomics approaches such as differential display RTPCR demonstrated up regulation of Rab Coupling Protein (RCP) in the endometria of antiprogestin treated animals, as compared to control animals (Patil et al, 2005). Interestingly RCP is known to be involved in the intracellular trafficking of integrins. There was no concomitant increase in the expressions of Rab4 and Rab11- proteins known to interact with RCP, suggesting impairment in the expression of specific components of intracellular trafficking pathways. These studies suggested that the blockade of progesterone action in endometrium may alter intracellular trafficking and this in turn could be responsible for the altered distribution of cell surface molecules such as integrins on endometrium. This could be one of the reasons for the incompetence of endometrium for implantation in antiprogestin treated animals. Further, 2D proteomics coupled with MALDI-TOF-TOF analysis revealed differential expression of two reticuloplasmins- endoplasmic reticulum resident proteins such as calreticulin and protein disulfide isomerase in bonnet monkeys rendered infertile with antiprogestin. Interestingly, calreticulin was also found to be less abundant in the 2D endometrial tissue protein map of mid-secretory phase as compared to that of the proliferative phase in humans (Parmar et al, 2008a). This suggested that the expression of calreticulin is downregulated in endometrial tissues during progesterone dominant phase as compared to estrogen dominant phase, interestingly, expressions of these two proteins were also increased in endometrium during very early stages of pregnancy. It may be mentioned here that endometrial estradiol receptor alpha, an estrogen regulated gene also showed increased expression during early stages of pregnancy (Rosario et al, 2008). This suggested that the expression of these proteins is positively regulated by estradiol in vivo. Our in vitro studies also suggested that estradiol positively regulates the expression of calreticulin whereas progesterone is somewhat inhibitory to the expression of calreticulin. In addition to calreticulin? Chain of fibrinogen, adenylate kinase isoenzyme 5, transferrin, annexin V, alpha-1-antitrypsin (AAT), creatine kinase and peroxidoxin 6 were also found to be differentially expressed in endometrium during the progesterone dominant phase as compared to that in the estrogen dominant phase in humans. Further similar studies on uterine fluid samples revealed higher expression of AAT and apolipoproteins during the progesterone dominant phase or mid-secretory phase as compared to that in the proliferative phase of cycle in healthy fertile regularly cycling women (Parmar et al, 2008b).Conclusion: These studies collectively led to identification of several factors in endometrium, which are either positively or negatively regulated by progesterone. This knowledge will help in the construction of progesterone regulated functional networks, which can be targeted for contraception or for infertility management.

Objective: In spite of the good sperm and embryo response to cryopreservation programs, oocyte response (post-thaw maturation and fertility) to cryopreservation procedures is very poor. Special cellular status such as very sensitive cytoskeletal system, early release of cortical granules and pre-fertilization zona hardening and changes in position and structures of ZP glycoproteins are the consequences of freezing procedures on the oocytes. However, matured oocytes can tolerate the freezing procedures better than immature oocytes. Cytoskeleton stabilizer compounds, like Cytochalasin D and B, improved the vitrification outcome mature and immature oocytes. Taxol is a cytoskeleton stabilizer and used for human immature, bovine and murine mature oocyte vitrification, successfully. The aim of this study is to investigate the effects of taxol on immature bovine oocyte vitrification.Materials and Methods: Material and Methods Indigenous bovine ovaries transported to the Lab in the physiologic saline contained penicillin streptomycin in 39oC. Cumulus Oocyte Complexes (COC) aspirated form follicles (2-8 mm) and their qualification assessed in the medium TCM supplemented with 10%FCS. Grade A oocytes subjected to five groups of experiments: Control (n=108): oocytes with the routine IVM procedure, CRP (n=51): oocytes which exposed to the vitrification solutions without vitrification and subjected to IVM procedure, Tax (n=51): oocytes which exposed to Taxol for 15 min then transferred to IVM medium, OPS (n=50): oocytes which were vitrified, thawed and subjected to IVM procedure and OPS-Taxol (n=50): oocytes which exposed to the Taxol after 15 min, vitrified, thawed and transferred to the maturation medium. IVM medium was TCM 199 supplemented with 10mg FSH, 10mg LH, 10% Bovine follicular fluid and 5% FCS. During maturation, oocytes incubated in the atmosphere with 5% CO2, 95% humidity for 24 hours. After the period of maturation, COCs denuded and stained with the conventional procedure of aceto-orcein satin for evaluating nuclear maturation. Before vitrification, COCs exposed to three medium: HM: TCM 199+10% FCS, V1: HM+10% EG+ 10% DMSO and V2: HM+20% EG+ 20% DMSO+0.05m sucrose, for 5 min within each medium then loaded in the Open Pulled Straw (OPS) and plunged into the liquid nitrogen tank. At least after 48 hours of vitrification COCs thawed in the thawing media composed of T1: HM+0.25m sucrose and T2: HM+0.15m sucrose. COCs held within each media for 5 min and transferred to the IVM medium, finally. Taxol (Abetaxel®, ) with the maturation medium with dose of 1 nmol/ml. Percentage of matured oocytes were analysed with General Linear Model of SAS and means separated with the Tukey multiple comparison test. Data expressed as least square means with SEM.Results and Discussion: The percentage of matured oocyte was not different (p>0.05) between CRP (71.9±5.4) and Tax (73.1±6.8) groups compare to Control (80.5±6.8). Vitrification (9.1±6.2) significantly affected the maturation rate compare to control (p<0.05). The percentages of matured oocytes was higher (p.0>05) in Taxol treated vitrified oocytes compare to the oocytes which were not treated with Taxol.Conclusion: These results show the impact of vitrification on oocyte maturation of immature bovine oocytes with no significant improvement with pre-freeze Taxol treatment.

Objective: The aim of this study was to evaluate the optimal transplantation site for ovarian tissue fragments in murine hosts. We compared the transplantation to the back muscle (B) versus the kidney capsule (K) in a mouse allograft model.Materials and Methods: Hemi-ovaries from 12-day-old mice were allografted into B and K of bilaterally ovariectomized same strain recipients which had undergone gonadotrophin stimulation (n 5 15). Graft survival after 27 days, angiogenesis and follicle development were scored and compared to age-matched control ovaries (38-day old, n 5 5). The ability of oocytes to be fertilized was studied after IVF, ICSI and embryos were transferred to recipient mothers. Anti-mouse CD 311 antibody was used to evaluate neo-vascularization in grafts.Results: Primordial follicle survival was higher (p < 0.01) and vascular support was better (p < 0.01) in B- than in K-grafts. From 34 oocytes retrieved from B-grafts (15 metaphase I, of which 14 matured in vitro, and 19 collected at metaphase II), 18 morulae were obtained. Transfer of 12 embryos obtained by ICSI led to three live offspring, and transfer of six IVF embryos to another recipient mother yielded four offspring, one of which was born dead and one showed placental anomalies.Conclusion: Primordial follicle survival was higher (p<0.01) and vascular support was better (p < 0.01) in B- than in K-grafts. From 34 oocytes retrieved from B-grafts (15 metaphase I, of which 14 matured in vitro, and 19 collected at metaphase II), 18 morulae were obtained. Transfer of 12 embryos obtained by ICSI led to three live offspring, and transfer of six IVF embryos to another recipient mother yielded four offspring, one of which was born dead and one showed placental anomalies.

Egg sharing or oocyte sharing has different meanings: one is usage of a donor for two recipients that the recipients share the donated oocytes and compensation expenses. But the more popular definition of oocyte sharing is that an infertile woman undergoing assisted reproductive techniques gives half of her own oocytes to a recipient in return for subsidized expenses of fertility treatment. This paper focuses on the later definition and compares this procedure with oocyte donation from ethical, religious, social and legal perspective. The key results are as follows: oocyte sharing is more acceptable upon Islam, ethically it does not put a normal and fertile young woman under risk of fertility drugs, anesthesia and operations, socially it reduces the danger of “oocyte business” like changing the donor to vender, payment, brokers, arguments, advertisements, etc … and legally there is no difference between these two procedure and if donation is acceptable, so is the sharing program. There are two major concerns about egg sharing which are as follows: 1. maybe there are some psychological effect on donors who do not succeed to have a child but the recipient does. Although this effect was not reported in many researches and donors were always happy about what they did, but with complete anonymity it can be reduced. 2. There is a concern about the health of donor who is infertile and can have some health problems and advanced age which can be afforded by definition of some criteria for the suitable egg sharers.

Objective: Germ cell number during ovarian organogenesis is regulated through programmed cell death. We investigated the expression of germ cell-specific VASA protein, apoptosis-related proteins BAX and BCL2, and DNA fragmentation in human developing ovaries from gestation week 12 to term.Materials and Methods: Human foetal ovaries from 13 women undergoing spontaneous abortion were fixed, paraffin-embedded and processed for immunohistochemistry to analyse temporal and cellular localisation of VASA, BCL-2 and BAX, and to detect apoptosis by TUNEL assay.Results: VASA showed a differential pattern of expression throughout differentiation and proliferative phase, prophase I to finally associate to Balbiani's body in primordial and primary follicles. BCL-2 was detected from week 12 to 17 and became undetectable thereafter. Strong BAX signal was detected in oogonia and oocytes from week 12 to term. Low levels (£10%) of TUNEL positive germ cells were detectable throughout gestation with a higher incidence (around 20%) at 18-20 weeks.Conclusion: VASA were specifically expressed in germ cells and displayed a stage-specific intracellular localisation enabling to follow oogenesis throughout gestation. Apoptosis-inhibiting BCL-2 was associated to germ cell proliferative phase and prophase I while BAX remained positive throughout gestation. The highest incidence of apoptotic germ cells was coincident with the lack of detectable BCL-2 protein, and when primordial follicle formation became widespread.

Objective: The ability to reproduce and give birth to a child is an inherent part of life for most human beings. To see the next generation grow up is described as happiness and a driving force in life; thus infertility causes severe anxiety for infertile persons. Approximately 15% of all couples in the reproductive ages are involuntarily childless, and infertility is the third cause of divorce in Iran. The purpose of this study was to explore men’s experience of male infertility.Materials and Methods: Descriptive phenomenological method was used to determine these experiences. The data was collected by deep interviewing with 10 interfile men.Results: In related with infertility phenomenon we obtained four general concepts: 1. Personal anxiety. 2. Challenge with communications. 3. Effects of believers and religion. 4. Problems associated with treatment process.Conclusion: According to our emerged result in this study, it seems that all aspects of infertile men’s life are affected by infertility. Thus, designing and accomplishment of consultive and supportive programs play very important roles for giving better cares to infertile men.

Objective: To determine the relationship between sexual satisfaction, its changes, and marital satisfaction according to personal characteristics in 45-65 years old women in south of Tehran.Materials and Methods: This is a Cross sectional study that was done in 2006-2007. The valid and reliable (test re test) questioner was used. The questioner had two main parts: 1) Personal characteristics (age, couples’ age difference, education level, job, No. of children, and No. of children who lives with their parents…), 2) Sexual satisfaction, its change and marital satisfaction. (Visual scale was used) The samples used in this study consisted of 161 volunteer healthy women with healthy couples, who were chosen from the public centres of low socioeconomic district in south of Tehran. We exclude the women with University degree. Descriptive and interferential statistical method (ANOVA, Scheffe, Pearson correlation) were used. The final data were analysed and tabulated in 32 tables.Results: Referring to personal characteristics, we found remarkable points. The highest average of marital satisfaction was in 60-65, sexual satisfaction was in 45-49, and sexual satisfaction change (decrease) was in 55-60 age groups. There was only difference between marital satisfactions according to average of age group. (ANOVA, p=0.03) By using Scheffe test we found this difference between 45-49 and 50-54 years age group. Also we found correlation between marital satisfaction and sexual satisfaction (pearson correlation, r=+0.728) and between sexual satisfaction and its change. (r=-0.215).Conclusion: According to personal characteristics, we only found difference between age groups. We found during early years of post menopause, there was more marital and sexual unsatisfied group, which may be because of not still coping with the menopause situation. We suggest having more education and consultation program for this group. Also we suggest having the same study with the same goals on other socio economical group and men too, for comprising the results.

Objective: Despite the lack of empirical research on the role of spirituality in the lives of infertile women, scientific literature has identified religion and spirituality as significant coping resources throughout the life course which individuals rely on to gain control in their lives. This study explored how infertile women coped with infertility using their belief system.Materials and Methods: In a feminist grounded theory study 30 infertile women affiliated to different denominations of Christianity (Protestantism, Catholicism, Orthodoxy) and Islam (Shia and Sunni) were interviewed. Data were collected through semi structured in-depth interviews with volunteer infertile women in one Iranian and two UK fertility clinics and analyzed using Strauss and Corbin’s mode of grounded theory.Results: Religious infertile women using religious and spiritual problem-solving strategies played an important role in promoting their psychological fitness. They used a combination of both positive (benevolent religious reappraisal, belief in spiritual support, engagement in religious rituals, belief in miracle, religious surrendering, belief in timing, religious consciousness, religious helping and seeking support from clergy) and negative religious coping strategies (demonic reappraisal, discontent with God, discontent with clergy and spiritual irrelevance). Religious participants concurrently used some non-religious coping strategies (seeking friendship, looking for medical resources, not to think about, changing lifestyle and complementary therapies) in conjunction with religious coping strategies.Conclusion: Infertile women reported a variety of religious/spiritual coping strategies which were associated with adaptive health outcomes. Further scientific inquiry is required to investigate how religion and spirituality promote healthy adaptation to infertility and whether health professionals' attention to religious/spiritual coping could assist infertile women to be well adapted to infertility.

Objective: PCOS is associated with several metabolic complications. Physical appearance, menstrual abnormalities and difficulty conceiving are related to lower health-related quality of life scores in these patients. The aim of this study is to estimate the prevalence of depression in PCOS patients compared with controls.Materials and Methods: Women with PCOS (n=55) meeting the Rotterdam criteria for this syndrome, who referred to outpatient endocrine clinic in Qom, Iran, compared with control subjects (n=55) who selected from healthy women and age matched, from Dec.2007 to Sep.2008. Beck Depression Inventory (BDI) questionnaire was filled up for participants. Blood pressure, weight, height, waist circumference were measured for all subjects. PCOS patients were divided in two subgroups: Depressed and non depressed. LH, FSH, Testosterone, DHEA-S, prolactin, fasting plasma glucose, lipid profiles were measured only in PCOS subgroups.Results: PCOS patients had significant higher prevalence of depression compared with controls (60% vs. 23.6%).The prevalence of severe depression was 21.8% vs. 1.8% in PCOS and control respectively. Systolic blood pressure (BP), diastolic BP, body mass index and waist circumference were higher than controls (p<0.05). We did not find significant differences in hormonal, lipid and glucose profiles between depressed and nondepressed PCOS patients. Although the numbers of infertile women was higher in PCOS patients, but this variable had not statistically significant difference between depressed and nondepressed PCOS.Conclusion: The present study shows that women with PCOS have significantly increased risk of depression and they should be routinely screened and adequately treated for depressive disorders.

Objective: To compare the efficacy of using mild ovarian stimulation protocol and conventional stimulation protocol in IVF outcome.Materials and Methods: We randomized 200 women with regularly menstruation who were candidate for IVF. They had undergone stimulation with clomiphene citrate, gonadotropin and GnRH antagonist or gonadotropin and GnRH agonist.Results: There were no significant difference in mean age, cause of infertility, basal FSH, BMI, endometrial thickness on the HCG administration and clinical pregnancy rate in two groups. The number of recovered oocytes, embryos obtained, embryos transferred,, peak estradiol on the HCG administration and OHSS were significantly higher in agonist group. Significantly more patients in agonist group had embryo cryopreserved.Conclusion: The CC/ gonadotropin/ GnRH antagonist is an acceptable alternative protocol for IVF in patients with regularly menstruation.

Objective: To compare the efficacy of using mild ovarian stimulation protocol and conventional stimulation protocol in IVF outcome.Materials and Methods: we randomized 200 women with regularly menstruation who were candidate for IVF. They had undergone stimulation with clomiphene citrate, gonadotropin and GnRH antagonist or gonadotropin and GnRH agonist.Results: There were no significant difference in mean age, cause of infertility, basal FSH, BMI, endometrial thickness on the HCG administration and clinical pregnancy rate in two groups. The number of recovered oocytes, embryos obtained, embryos transferred, peak estradiol on the HCG administration and OHSS were significantly higher in agonist group. Significantly more patients in agonist group had embryo cryopreserved.Conclusion: The CC/ gonadotropin/ GnRH antagonist is an acceptable alternative protocol for IVF in patients with regularly menstruation.

Objective: To evaluate the predictive value of basal serum anti-müllerian hormone level and small antral follicle count for high ovarian response to controlled ovarian hyperstimulation.Materials and Methods: A total of 159 patients were prospectively included. Basal serum anti-müllerian hormone and small antral follicle count (2-6 mm) were measured.Results: Small antral follicle count and anti-mullerian hormone have similar predictive accuracy for high ovarian response with area under curve of 0.961 and 0.922, respectively. The sensitivity and specificity for prediction of high ovarian response were 89% and 92% for small antral follicle count and 93% and 78% for anti-müllerian hormone at the cutoff values of ³ 16 and ³ 34.5 pmol/l, respectively.Conclusion: Small antral follicle count and anti-müllerian hormone are equally accurate predictors of high ovarian response and facilitate determination of the optimal strategy for controlled ovarian hyperstimulation.

Objectives: This multicenter, randomized, single-blind study assessed the safety and efficacy of a resorbable hydrogel (‘Hydrogel’) for the reduction of post-operative adhesion formation following myomectomy.Materials and Methods: Women (n 5 71) who were undergoing laparoscopic (67.6%) or laparotomic myomectomy were randomized (2:1) to Hydrogel (sprayed over surgically treated areas prior to wound closure, n 5 48) or to control (standard care, n 5 23). Patients (38 Hydrogel, 20 control) returned 8-10 weeks later for a second look. Adhesions were graded using a modified American Fertility Society (mAFS) scoring method. The primary efficacy measure was the posterior uterus mAFS score.Results: For Hydrogel and control patients, respectively, Mean±SD mAFS scores were 0.5±1.4 and 0.0±0.0 at baseline, and 1.1±1.9 and 2.6±2.2 at the second look. Similarly, mean changes from baseline were 0.8±2.0 and 2.6±2.2 (P 5 0.01); 95% confidence intervals for these mean changes were (0.16 - 1.44) and (1.64 - 3.56). Adverse events were reported by 9.6 and 17.4% of Hydrogel and control patients, respectively. No intraabdominal infections or post-operative site infections were reported.Conclusion: This 71-patient study provides the first clinical evidence of the safety and efficacy of Hydrogel for the reduction of adhesions following myomectomy.

Objectives: The foeto-maternal relationship during pregnancy is controlled by a complex regulation of immuno-endocrine homeostasis where progesterone-dependent immunmodulation plays a key role. Due to stimulation by foetally derived antigens, lymphocytes of healthy, pregnant women express progesterone receptors and in the presence of progesterone produce a 34-kDa mediator protein, named the progesterone-induced blocking factor (PIBF). PIBF induces a Th2 dominant cytokine production via inhibiting STAT4 and activating STAT6, inhibits maternal NK activity which results in decreased cell-mediated response thus exert an anti-abortive effect. Though PIBF does not directly bind to IL-4 receptor-alpha, our aim was to determine the role of IL-4Ralpha in PIBF signalling. Furthermore, we investigated the effects of PIBF on the protein kinase C (PKC)/ Ca++ system which play a key role in Th1/Th2 differentiation.Materials and Methods: Confocal microscopy was used to detect the localization of IL-4Ralpha and PIBF receptor (PIBFR). To verify the involvement of IL-4Ralpha in PIBF signalling, IL-4Ralpha was silenced by oligonucleotides interfering with IL-4Ralpha mRNA. Assuming that the PIBF receptor is a GPI-anchored protein, PIBF-induced phosphorylation of STAT6 was tested in phosphatidylinositol-specific phospholipase C (PI-PLC) digested cells by Western blotting. The hypothesis that PIBF receptors float in glycosphyngolipid-cholesterol rafts was tested by depletion of cholesterol using methyl-b-cyclodextrin (MbCD). Proteins from PIBF-treated cells were reacted on Western blots with phospho-specific antibodies recognizing different PKC izoforms. Intracellular free calcium was measured by flow cytometry.Results: Labelling of the IL-4Ralpha and simultaneous activation of the PIBF receptor with a FITC-labelled ligand revealed co-capping of the two binding sites. In IL-4Ralpha deficient cells the STAT6 activating effect of PIBF was markedly reduced. After PI-PLC treatment PIBF did not induce STAT6 phosphorylation while IL-4 retained the same effect. In MbCD treated cells neither PIBF, nor IL-4 were able to Tyr-phosphorylate STAT6, suggesting that not only the PIBFR but also the IL-4Ralpha is enriched in lipid rafts. Both IL-4 and PIBF induced PKC phosphorylation which was abrogated by anti- IL-4Ralpha or anti-PIBF IgG pre-treatment. PIBF treatment did not alter intracellular Ca++-levels. Inhibition of PKCzeta or PKCtheta phosphorylation, but not that of PKCalpha/beta resulted in the loss of STAT6 and JAK1 phosphorylation by PIBF.Conclusion: Our findings suggest the existence of a novel type of IL-4R, composed of the alpha-chain of IL-4R and the PIBFR. The PIBFR is a GPI-anchored protein that lacks cytoplasmic tail thus it uses the intracellular domain of IL-4Ralpha for signalling. Upon ligand-binding, the PIBFR enriched in lipid rafts forms a complex with the IL-4Ralpha subunit and activates the JAK1/STAT6 pathway. PIBF phosphorylates PKC via binding to the IL-4R, without affecting intracellular Ca++. Phosphorylation of PKCzeta and PKCtheta is required for JAK1 and STAT6 activation, whereas PKCalpha/beta is not involved. These findings explain the mechanism by which PIBF supports a Th2 dominant cytokine pattern.

Objective: M-CSF is a cytokine mediating the growth, proliferation and differentiation of various cell types including macrophages, trophoblast cells, and osteoclasts. It plays an important role in immunity reproduction, follicle development and ovulation. In the present study, we describe the important role of the changes in serum M-CSF levels during the menstrual cycle, in the process of follicular maturation, ovulation, implantation, pregnancy and their response to ovarian stimulation with recombinant (r) FSH.Materials and Methods: From an original sample of 95 IVF/ICSI patients (mean age=33.8±5.4), serum and follicular fluid (FF) were collected on the day of follicular puncture (FP). The M-CSF levels were measured by ELISA technique. These patients were divided into two groups as follows: In Group 1 patients with the aetiology of tubal or male factor infertility were analysed for: a) correlation between serum and FF with respect to M-CSF and correlation between M-CSF and estradiol (E2) in serum; and b) comparison of M-CSF level in serum in response to ovarian stimulation and comparison of M-CSF level in serum between pregnant and non-pregnant patients. In Group 2: Patients (n=23) were monitored throughout the menstrual cycle until 4 weeks after embryo transfer. In this group, M-CSF levels in serum were analysed throughout the different ovarian cycle phases and gestation.Results: M-CSF levels in FF were higher than in serum (p=0.01). M-CSF levels in serum increased from low, through moderate, to high response patients (p=0.001); pregnancy rates were 11.5%, 22.5% and 51.7%, respectively. M-CSF in serum increased throughout stimulation until the day of oocyte retrieval (p=0.006) and decreased until embryo transfer (ET, p=0.03). In the post-retrieval days, from the day of ET, through implantation, to the day of confirmation of pregnancy, the M-CSF levels of those patients who became pregnant (n=13) increased significantly (p=0.03) and reached their highest level. After implantation the M-CSF levels decreased slightly and reached a plateau during gestation.Conclusion: our data show that M-CSF is involved in follicle development and ovulation and plays an important role in the maintenance of pregnancy. It could also be a predictor of embryo implantation for IVF outcome.

Objective: The aim of this study was to evaluate the effects of myo-inositol on the lipid pattern and insulin sensitivity in hirsute women.Materials and Methods: 129 hirsute women were enrolled to the first Institute of Obstetrics and Gynecology (University La Sapienza, Rome, Italy) and evaluated at baseline and after receiving six months of myo-inositol therapy (4 gr/day). Body Mass Index, hirsutism score, serum levels of total cholesterol, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, triglycerides, apolipoprotein B, lipoprotein (a), serum adrenal and ovarian androgens, fasting glucose and insulin concentrations were evaluated.Results: The hirsutism score decreased after therapy (p value <0.001). Total androgens evaluated decreased, as a decrement of FSH and LH levels, while E2 levels increased. There was a slight decrement of total cholesterol levels but it isn’t statistically important, an increment of HDL cholesterol levels and a decrement of LDL cholesterol levels. No significant changes were observed in serum triglyceride, apolipoprotein B and lipoprotein (a) concentrations. Fasting insulin levels and insulin resistance analyzed by homeostasis model assessment were reduced significantly after therapy.Conclusion: Administration of oral myo-inositol significantly reduced the hirsutism score and hyperandrogenism and ameliorates the abnormal metabolic profile of women with hirsutism.

Objective: The expression of adrenomedullin (ADM) in the reproductive system of the female rat and its effect on the secretion of estradiol and progesterone was studied.Materials and Methods: In adult Sprague-Dawley rats, gene expression of ADM and its receptor components in the ovarian follicles, oviduct was analyzed by real-time PCR (qPCR). Ovarian follicles and corpora lutea isolated from stimulated immature rats were cultured to study the effects ADM on in vitro basal, FSH-stimulated estradiol secretion or eCG stimulated progesterone secretion.Results: Ovarian ADM and Adm mRNA levels were decreased at estrus, whereas oviductal Adm mRNA levels were low at proestrus. Both tissues were shown to coexpress mRNAs encoding the calcitonin receptor-like receptor and receptor activity-modifying protein 1 (Ramp 1), Ramp 2, and Ramp 3. Gel filtration chromatography of ovarian extracts showed two peaks, with the predominant one eluting at the position of authentic rat ADM (1-50) at estrus and at the position of ADM precursor at diestrus. Positive ADM immunostaining was localized in the granulosa and theca cells of the follicle and corpora lutea of the ovary. ADM inhibited FSH-induced estradiol secretion in 2-day-old follicles and also suppressed eCG-stimulated progesterone release in the corpora lutea was abolished by calcitonin gene-related peptide (8-37) and ADM (22-52), respectively.Conclusion: The presence of ADM and the gene expression of ADM and its receptor components in the female reproductive system suggest a paracine effect of ADM on ovarian steroidogenesis.

Objective: Mutational events may be an indirect effect on genome stability which is transmitted through the germ line of chemically or physically exposed parents to their offspring. The consequences of germ cell mutations in subsequent generations include genetically determined phenotypic alterations without signs of illness, or reduction in fertility, or embryonic or prenatal death, more or less severe congenital malformations, or genetic diseases with various degrees of health impairment. In this study, the effect of induction of DNA damage during spermatogenesis cycle and preovulatory stage oocyte on the frequency of chromosomal abnormalities and micronuclei formation in preimplantation embryos generated by damaged sperm or oocyte after exposure to gamma rays in the presence or absence of vitamins E and C is investigated.Materials and Methods: DNA damage was induced in male NMRI mice using gamma-rays, and then mated with non irradiated super-ovulated female mice in 6 successive weeks after irradiation in a weekly interval. In experiments involving irradiation of both male and female mice, irradiated male mice for 6 weeks post-irradiation were mated with female mice irradiated after induction of super-ovulation. To study the effects of vitamins E and C on the radiation induced DNA damage and consequently in the chromosomal abnormalities generated in preimplantation embryos, vitamin E at a concentration of 200 mg/kg and vitamin E at 100 mg/kg was administered to mice interaperitoneally 1 hour before exposing to radiation. Standard methods were used to prepare slides from pre-embryos for chromosome and micronuclei study.Results: The rate of both aneuploidy and MN observed in embryos generated from irradiated male compared to control group dramatically increased (p<0.01). Frequency of aneuploidy and MN in embryos generated by mating both male and female irradiated mice was higher than that observed for those embryos generated by irradiated male mice alone. Cells at early spermatogenetic cycle were more sensitive to radiation and led to higher frequency of aneuploidy and micronuclei in preimplantation embryos. Exposure of male and female animals to gamma irradiation in the presence of vitamins E and C led to a considerable reduction in both chromosomal aneuploidy and micronuclei in generated preimplantation embryos by irradiated parents. Reducing factor for vitamin E was found about 2 and for vitamin C about 3. Also a direct correlation between aneuploidy and micronuclei formation was observed.Conclusion: Results indicate that induction of DNA damage in gonads during spermatogenesis and pre-ovulatory stage oocytes may lead to unstable chromosomal aberrations and probably stable chromosomal abnormalities affecting pairing and disjunction of chromosomes in successive pre-implantation embryos expressed as aneuploidy and micronuclei. These types of chromosomal alterations may lead to impaired embryonic and fetal developments. Administration of vitamins E and C before irradiation effectively reduced the frequency of chromosomal abnormalities. The way these vitamins reduces genotoxic effects of radiation might be via radical scavenging mechanism or antioxidative effects. Higher dose reduction factor observed for embryos generated after vitamin C treatment might be due to water soluble nature of this vitamin or its direct involvement in DNA repair. This observation might have a great impact for cancer patients under radiotherapy, occupationally exposed individuals to physical and chemical DNA damage inducing agents and residents of high natural background radiation area who experience impaired gametogenesis and fertility.

During the last decade the Toll Like Receptors (TLR) were found to be a major part of innate immune system. Several reports have demonstrated the existence of TLRs in different tissues and organs. However, little has been done to identify TLRs in the female reproductive tract (FRT). In addition little information existed regarding variation in TLRs in FRT during the menstrual cycle and also the influences of sex hormones on TLRs expression and function in this tract.The distribution of TLR7-10 protein was detected by immunostaining in endometrium. Also RT-PCR was used to show the existence of TLR genes in human fallopian tube, endometrial tissue and OE-E6/E7 cells as a reliable model of fallopian tube epithelial cell. Q-PCR analysis was used to investigate relative expression of these genes during the menstrual cycle in human endometrium. In addition, separate and synergistic effects of sex hormones on TLRs were tested in OE-E6/E7 cells by using Q-PCR and ELISA. To confirm the specific effects of sex hormones in the study, antagonists against estradiol and progesterone were used.The results showed that TLR7-10 proteins were present in endometrial epithelium and stroma. TLR1-10 genes were expressed in human fallopian tube and endometrial tissue. The mean relative expression of TLR genes was significantly higher during the secretory phase of the menstrual cycle in endometrium. In addition, it was demonstrated that TLR 1-6 genes and proteins were expressed in OE-E6/E7 cell line. Our data clearly showed that Estrogen has no effect on the expression of TLRs except TLR1 in OE-E6/E7 cells. In contrast, progesterone had an inhibitory effect on the expression of TLR1-4 genes in this cell line. Moreover, we proved that the production of IL-6 was significantly increased in the presence of TLR3 ligand (poly (I:C)), and both sex hormones had a suppressive and biphasic effect on the production of IL-6 in the presence and absence of poly (I:C). Our results also suggested that the estrogen receptor β and nuclear progesterone receptor B are likely to mediate the hormonal regulation of TLR3 as these two receptors are the two main estrogen and progesterone receptors in OE-E6/E7.These results imply a potential variation in the expression and function of TLRs in FRT when different levels of sex hormones are present during different stages of the menstrual cycle. Clinically, Limited success in dealing with infertility issues and protection against sexually transmitted disease demonstrate the need for a greater understanding of the regulation of immune system in the female reproductive tract. Studying the function of TLRs in the female reproductive tract presents an exciting opportunity to further understand the regulation of innate immune system in the female reproductive tract. It seems sex hormones regulate the function and expression of TLRs in the female reproductive tract and therefore influence/regulate innate and adaptive immune function in this tract to protect against potential pathogens while providing an environment that supports an allogeneic foetus. How TLR function and expression is exactly regulated in reproductive tract by sex hormones would be a challenging but fruitful area of future research.

Leiomyomata uteri are the most frequent myometrial disorders and the most common pelvic tumor in women. Although most women with uterine leiomyomas do not seek therapy, 20% to 40% of women in the reproductive age do have significant enough symptoms caused by these fibroids to cause the woman to seek and warrant therapy.Since 1995 uterine fibroid embolization (UFE) or uterine artery embolization (UAE) was originally devised to reduce pelvic bleeding due to postpartum hemorrhage. It has been introduced as a treatment for symptomatic uterine myomas sparing the uterus.Without a good blood supply, it has been shown fibroids will decrease in size between 30% and 50% and decrease in symptomatology.Initial studies have shown that UAE can improve menorrhagia in 90% of patients at 1 year after therapy. On average, the volume of the fibroids decrease by 30%–60% and the associated symptoms (of mass effect) are successfully treated in 71% of patients.Complications include postprocedure pain and postembolization syndrome possibly related to the release of cytokines and toxins from the ischemic tissue.Vascular anastomotic communications between the uterine and ovarian arteries provide a route by which embolization materials can affect the ovarian blood supply and ovarian function, either permanently or temporarily. So Currently, UAE is not recommended as the first line of therapy in patients with infertility presumed to be caused by fibroids.Magnetic resonance imaging–guided focused ultrasound surgery (MRIgFUS) is another groundbreaking minimally invasive alternative to surgery for fibroids. Focused ultrasound causes local tissue thermal coagulation, ablates the target fibroid, and allows preservation of uterine function. It is a feasible and safe outpatient procedure that does not require hospitalization.The procedure begins with the delivery of low-power (50–100 watt) sonication, with real-time thermometry acquired simultaneously. We will discuss these two techniques in detail and share our experience.

Varicocele is the commonest curable cause of male infertility.According to the statistical data collected from literature review there is 10-25% risk of recurrence after routine methods of varicocele operation.If a surgeon is certain about accuracy of the surgical method, probable cause of recurrence would be presence of collaterals and anatomical variations, so embolization would be the method of choice in case of recurrence.

The traditional method of screening for Down’s syndrome has been maternal age where amniocentesis or chorionic villous sampling is offered to women aged 35 years or more. This results in the need for an invasive test in 15-20% of pregnant women with a detection of less than half of the fetuses with Donw’s syndrome, because the majorly of affected fetuses come from the younger age group.A more effective method of screening is based in the combination of:● Maternal age● A maternal blood sample for the measurement of the placental products of free b-hCG and PAPP-A● An ultrasound scan at 11-13 weeks:○ to measure the collection of fluid behind the fetal neck (nuchal translucency)○ to examine the fetal nose and palate○ to measure the fetal heart rate○ to assess the flow of blood across the tricuspid valve of the fetal heart and the ductus venosusThis new method of screening reduces dramatically the number of women requiring an invasive test from about 20% to less than 3% and at the same time increases the detection rate of Down’s syndrome and other major chromosomal abnormalities form less than 50% to more than 95%.

Objective: Increased nitric oxide in the spermatic veins of men affected by varicocele has been reported. NO plays an important role in the varicocele-induced decrease of seminal quality. iNOS activity may play a role in the testicular dysfunction associated with varicocele during adolescence. iNOS activity was slightly increased in the leydig cells of varicocelized rats. It is a clinical case that inhibitors of NOS have succeeded in enhancing motility. Aminoguanidine is an inhibitor of iNOS. The objective of this study was to investigate the role of AG in efficiency of sperm parameters in varicocelized rat.Materials and Methods: Twenty four wistar male rats divided into four groups. The group A and B underwent a left experimental varicocele. In group C (sham group), rats underwent a similar procedure but without any change on spermatic vein. Group D referred to as control. Animal in group A were killed 10 weeks after the operation. Epididymal sperm parameters were evaluated. Group B received 50 mg/kg AG intraperitoneally daily for ten weeks.Results: Sperm parameters decreased significantly in the group A and the group B in comparison with group C and D (p£0.05). The sperm parameters increased in the group B that received AG in comparison with the group A (p£0.01). Group C (sham) did not show any significant alteration in sperm parameters in comparison with the control group.Conclusion: These findings suggest that iNOS is an important mediator in the pathogenesis of varicocele. This study may support the concept that the use of iNOS inhibitor in infertile men with varicocele may be useful.

Objective: The aims of this case-control study were to identify the seminal fluid patterns in Iraqi men with infertility as well as in fertile controls .Also to demonstrate the types of serum hormones (Follice Stimulating Hormone (FSH), Leutinizing (LH), Testosterone and Prolactin) abnormalities in the study groups.Materials and Methods: 81 Iraqi men with infertility and 30 fertile men who fulfilled the selection criteria to whom seminal fluid analysis (SFA) was performed according to WHO method. The patients group was subdivided by sperm concentration into azoospermic, oligoasthenozoospermic and oligozoospermic subgroups. Serum levels of the hormones (Testosterone, FSH, LH and Prolactin) were measured for patients and controls using ELIZA immunoassays. Seminal fluid analysis parameters mean levels and serum hormones levels were compared for the groups using Analysis of Variance test.Results: Iraqi infertile men showed lower values for SFA parameters than did the controls. Patients with azoospermia showed the most remarkable hormonal abnormalities especially in the levels of serum FSH and Testosterone. Patients also demonstrated multiple abnormalities in seminal fluid parameters. There was a significant difference in the serum sex hormones levels between the patients and controls groups and among the infertile men subgroups.Conclusion: Seminal fluid abnormalities among Iraqi patients with male infertility are multi-components. Hormonal profiles for this patient do not follow a single pattern. Patients with low sperm concentration and especially those with azoospermia are those that most likely will get benefit from hormonal assays. Serum FSH and Testosterone are the best 2 hormones for initial male infertility evaluation.

Objective: Freeze and thaw have some detrimental effects on sperms. Molecular changes on the sperm surface can impact on fertility rate. We tested wheat germ agglutinin (WGA), peanut agglutinin (PNA) and Dolichos biflorus (DBA) to detect surface glycoconjugates before and after freezing.Materials and Methods: For this purpose, 45 healthy semen samples were frozen and thawed and sperm smears were prepared before and after freezing and thawing. The smears were stained with the lectins and also with acridin orange. The smears were studied by fluorescents microscopy and the intensities of the reactions to lectins were measured by image analyses software.Results: The results indicated that the reactions of 46.67%, 34.09% and 73.34% of the specimens modified to PNA, WGA and DBA lectins respectively, after freezing and thawing. The cryopreservation caused both increase and decrease of the intensities of the reactions. It means that there are various mechanisms that impact on the carbohydrate contents of the sperm surface.Conclusion: We conclude that cryopreservation affected the surface glycoconjugates at least in a subset of spermatozoa. These results might improve future application of sperm banking techniques.

Objective: In recent years concerns have been raised about the decrease of male infertility from exposure to environmental pollution such as heavy metals. So, developmental effects of lead acetate were studied on seminiferous tubules structure during postnatal development in male Wistar rats.Materials and Methods: The male pups randomly divided in control and experiment groups. The mothers of experiment groups received 100, 200 and 300mg/L/day lead acetate via drinking water during lactation. The testes of pups removed their weights were recorded and fixed in Bouin’s solution. Then, the volumes of testes were measured by Cavalieri method. For microscopic studies, following tissue processing, 5µm sections were stained with haematoxylin-eosin.Results: Macroscopic results showed that the means of testis weight of experiment groups increase during early and decrease at the late of postnatal development in comparison with control group. Microscopic results indicated that decreasing of germinal epithelium height and presence of vacuoles in germinal epithelium comparison with control group. Also, the means of testis volume, relative volumes of seminiferous tubules and germinal epithelium significantly decreased in experiment groups during all stage of postnatal development (p<0.05). Moreover, the means of seminiferous diameter and epithelium height significantly decreased in experiment groups during all stages of postnatal development (p<0.05).Conclusion: Present study indicate that lead acetate could affect postnatal development of seminiferous tubules in rats and decrease spermatogenic activity of adult Wistar rats that have been exposured to it during neonatal period.

Objective: 1,4Dihydro 2,6Dimethyl 4(4.Nitrophenyl) 3,5Pyridinedicarboxylic acid Dimethyl ester (DDNPD) is a new analogous of Nifedipine that has no effect on calcium channels of cardiovascular system. Because Nifedipine can reduce fertility in males (through effect on calcium channels on sperm membrane and inhibition of acrosomal reaction), this study was done to evaluate the effect of DDNPD on reproductive system physiology and fertility in male rats.Materials and Methods: In this research, DDNPD was injected 10mg/kg twice a day subcutaneously to the test group of adult male rats for 50 days. Control group treated with propylene glycol 1 mg/kg twice a day in the same way.Results: In the assessment of some fertility indices, we saw a significant decrease in motility and viability rate of sperm (p<0.001), the epididymal sperm reserve rate (p<0.001) and fertility rate (p<0.001) in the test group but there was no difference in testosterone level between the groups.Conclusion: These results are conclusive of post testicular antifertility effect of DDNPD if injected 10 mg/kg for 50 days to male adult rats.

Objective: Testicular torsion is the twisting of the spermatic cord, which cuts off the blood supply to the testicle. The main pathophysiology of testicular torsion is ischemia-reperfusion injury caused by the twisted spermatic cord and its release. Reperfusion component typically involves the generation of the reactive oxygen species with the return of blood flow after the ischemia. The aim of the present study was to investigate the effects of dopamine, a vasodilator, and vitamin C, an antioxidant, on ischemia reperfusion injury due to testicular torsion-detorsion (T/D).Materials and Methods: Thirty adult male NMRI rats were divided randomly into six groups each containing 5 rats. Testicular ischemia was achieved by twisting the left testis 720° clockwise for 4 hour, and then reperfusion by detorsion. Group 1 was for the basal values. Group 2 was designed as sham-operated without any T/D. Group 3 had only T/D. Group 4 received vitamin C (100 mg/kg, i.p) just after detorsion. Group 5 received dopamine (0.01 mg/kg, i.p) after 4 hours T/D and group 6 received combination of vitamin C and dopamine. The animals in each group underwent orchiectomy 48 hours after detorsion and the testicular tissues were fixed in 10% formaldehyde for histopathological examinations.Results: In groups 1 and 2, microscopic features were normal. In group 3, all of the seminiferous tubules were damaged. In some tubules only the basement membrane was seen. Pyknosis of spermatocytes nuclei was prominent. Germ cells and sertoli cells were decreased and underwent degeneration and necrosis. Interestingly in group 4, only a few degenerative changes (6.8%) were observed in tubules. In group 5, approximately 90% of tubules were underwent full or partial degeneration and in group 6 only few normal tubules were found (2.2%).Conclusion: Vitamin C administration exerts a beneficial effect on testicular ischemia-reperfusion injury but not dopamine or even dopamine and vitamin C combination.

Objective: Phthalate esters extensively used as plasticizers in the plastic industry have been shown to cause reproductive toxicity in both developing and adult animals. So, present study was aimed to determine quantitative effects of di-(2-ethylhexyl)-phthalate (DEHP) on postnatal development of rat testis.Materials and Methods: For this purpose, pregnant females rat were orally exposed to various doses (0, 100, and 500 mg/kg per day) of DEHP during gestation and lactation. Testes of offspring were removed at 7, 14, 28, 60 and 90 days after birth, their weight recorded and fixed in bouin’s solution. Then, the volumes of testes were measured by Cavalieri method. For microscopic studies, following tissue processing, 5 mm sections were stained with haematoxylin-eosin.Results: The results were showed that the means of testis weight decreased significantly (p<0.05) in low dose group at 90 days after birth and in high dose group from 28 days after birth in compared with control group. Total volume of testis at 90 days after birth in both low and high dose groups reduced significantly (p<0.05) 14.54 and 10.03 percent in compared with control group, respectively. Relative volume of seminiferous tubules showed significant reduction from 14 days after birth in both experiment groups. Seminiferous tubules diameter at 90 days after birth decreased days significantly (p<0.05) 15.20 and 21.57 percent in compared with control group, respectively.Conclusion: Present study determines degree of quantitative variations in testicular parameters during postnatal development in Wistar rats exposed to di-(2-ethylhexyl)-phthalate (DEHP).

Objective: The benefit effect of L-carnitine is proposed for treatment of various dysfunctions of sperm in infertile men. So, in the present study, effect of L-carnitine supplement on sperm parameters in men with idiopathic infertility is evaluated.Materials and Methods: Thirty infertile men, ages 20 to 40 years, with the following baseline sperm selection criteria (sperm count <66.6 × 106, motility < 30%, viability <60%, normal morphology <35%) completed this study. Patients underwent L-carnitine therapy 3 g/day; the study design was 3 and 6 months of therapy. Semen analysis was performed according to World Health Organization guidelines. Sperm parameters included liquefaction, pH, volume, sperm count, motility, viability and normal morphology.Results: The results showed that L-carnitine supplementation increased sperm count, motility, viability and normal morphogy and pregnancy rate after 3 months (p<0.01) and 6 months (p<0.001), significantly. Also, L-carnitine supplementation increased sperm motility and viability in idiophatic infertile men after 3 and 6 months (p<0.001). On the other hand, 5 couples became pregnant during the study.Conclusion: The present study indicated that L-carnitine supplementation is an appropriate drug for treatment men with idiopathic infertility.

Objective: Cigarette smoking is associated with impairment of testicular function. Nicotine is a major component of cigarette smoke. In the present study we aimed to demonstrate the effect of various doses of nicotine on sperm parameters and quality of spermatogenesis in adult mice.Materials and Methods: Male adult NMRI mice were divided into four groups. 1) Control group were received normal salin. The animals in experimental groups of 2, 3 and 4, were received nicotine at the doses of 0.1, 0.2 and 0.4 mg/100 g. BW respectively. Both nicotine and vehicle were administered intraperitoneally for 21 days. Evaluations were made by determining Johnson's score, epididymal sperm content, motility and morphology, leydig cells volume and serum concentration of testosterone. Statistical analysis was performed by ANOVA test.Results: Nicotine caused a significant reduction in quality of spermatogenesis, sperm content, leydig cell volume and serum concentration of testosterone in groups of 3 and 4( p<0.05). Administration of nicotine in dose of 0.1 mg/100g. BW did not affect on the quality of spermatogenesis and sperm content. However both sperm morphology and sperm motility (p<0.05) and testosterone level (p<0.001) were decreased significantly.Conclusion: These findings suggest that administration of nicotine for 21 days leads to a deficiency in sperm characteristics, maturation of spermatogenesis and secretory dysfunction of the leydig cells. The adverse effects of nicotine are increased in a dose dependent man\ner.

Objective: survey the Varicoceles's rate and intensity in patients who are referred To the Fatemeh Zahra(s) Infertility center.Materials and Methods: In this field of descriptive study, the 713 file’s information of infertile couple that referred to the Fatemeh Zahra(s) infertility center in 1387 is surveyed. The information which related to fertility's action is registered. The medical examination of Varicoceles was done by urologist with physical medical examination way, and at least results were surveyed by chi-square test.Results: The frequencies of the first and second infertility were in order 73.5% and 26.5%.The average of men’s age in group with first infertility 9.72 years.±6.18 years and in group whit second infertility 34.69±30.2 Varicoceles frequencies was totally 34% and in men with the first and second infertility were ordered 35%and 32% that didn't show meaningful difference. The most frequency of Varicoceles intensity, grade II by 57% and left Varicoceles by 67%. The most percent of Varicoceles is in under 40-year-old group with 34%.Conclusion: with surveying the frequency of Varicoceles in this center. We can understand the rage of infertility patients in area .and evaluate according to age, kind of infertility and intensity of it to provide next detective and curative couple's programs, even to prevent before of the marriage.

Objective: This study was performed to investigate the suppressive effect of testosterone enantate on male germ cells apoptosis induced by busulfan in rats.Materials and Methods: Sprague- Dawley rats were divided into 4 groups: 1. a control group given cotton seed oil, 2. a group given two doses of busulfan 10 mg/kg intraperitoneally, 21 days apart, 3. a group given two doses of testosterone enantate 8mg/kg subcutaneously, 21 days apart, 4. a group given both testosterone anantate and busulfan. Evaluations were made by determining johnsen’s score and apoptosis were assayed by terminal- deoxynucleotidyl- transferase-mediated dutp nick end labeling (TUNEL).Results: Recovery status and johnsen’s score in group 4(busulfan + testosterone) were significantly higher than group 2(busulfan) 7.4±0.20 VS 4.1±0.22, TUNEL positive cells were significantly more numerous in busulfan treated group than control. While testosterone significantly attenuated morphological changes and germ cell apoptosis in testis damaged by busulfan.Conclusion: These results suggest testosterone enantate have chemoprotective effect on male germ cells against busulfan-induced toxicity partly by decreasing of apoptosis.

Objective: Celecoxib is a nonstroidal anti-inflammatory drug .This is a selectiveCox-2 inhibitor.Nowadays this drug uses as an analgesic, antipyretic and anti-inflammatory agent frequently. In some patients that used celecoxib for a long time, unusual effect of this drug may be seen. The goal of this survey is assess the effect of this drug on male-reproductive system functions.Materials and Methods: This survey done for study of effect of celecoxib on rat reproductive system, specially on spermatogenasis and the level of blood testosterone hormone. In this manner histologic studies and measuring of weight (testis, prostate, seminal vesicle and epidydimis) and the level of blood testestron are done. 50 rat with 200-230 gr. weight selected and compared in 5 groups.control group (no drug given), sham group (solvent drug: Di- methyl sulfoxide), 3 cases group (orally celecoxib 10, 20 and 40 mg/kg given daily) for 15 days. In the end of 15 days heart blood sampling for measuring serum testestron level accomplished after that reproductive systems separated and prepared for histologic study.Results: Result showed no significant differences in mean weight of body testis, epidydimis and seminal vesicle in control and case groups. But significant differences are seen in the mean weight of prostate per body weight in case group (40 mg/kg) in compared with control group. That is due to antimalignant effect of celecoxib on prostate. No differences seem between control and case groups in arrangement mode, nuclei shape and cytoplasm in histologic examination in spermatogonia and primary spermatocytes in transverse section of seminiferous tubules celecoxib in case group (40 mg/kg) can decrease lydige cells number, that is due to inhibited prostaglandin synthesise, for the result of cox-2 inhibitor and decreased level of testosterone hormone.Conclusion: It looks that number of sertoli cells in control and case groups are differences. So that in case group (40 mg/kg) number of sertoli cells decreased due to decrease testestron level. This can cause production of abnormal sperms. In the survey can conclude that use of high doses of celecoxib can decreased size and number of lydig cells and this is cause of decreased testosterone hormone.

Objective: “Glutathione, is a tripeptide” and known as GSH, just might be one of the most extraordinart overall health - boosters of modern nutritional medicine. GSH may help with everything from heightened immune system functioning to overcoming addictions to anti-aging and objective.” Many people are aware that sperm counts have dropped by half in the last 50 years, and those modern men have 20 percent less semen volume than their fathers. Persistent organic pollutants (POPs) and endocrine-disrupting chemicals from normal, everyday plastics are known to cause reproductive damage; Damage tosperm caused by exposure to common chemicals like alcohol and pesticides in food has been linked to lowered intelligence and behavioral disorders in children. Studies show that anti-oxidant supplementation - glutathione in particular - can improve sperm quality, and possibly increase your chances of conceiving.Materials and Methods: This is review article.Results: Mammalian spermatozoa are coated by a membrane rich in polyunsaturated fatty acids . These fatty acids are extremely susceptible to oxidative damage by free radicals or Reactive Oxygen Species (ROS) by a process called lipid peroxidation (LPO). Lipid peroxidation damages the sperm cell membrance . It is considered to be the key mechanism of ROS- induced sperm damage and leads to * Loss of sperm motility * Abnormal sperm morphlogy * Reduced capacity for oocyte penetration * Infertility To protect sperm from damage, the body depends on powerful antioxidant enzymes in the body such as superoxide dismutase (SOD), catalase, and glutathione/reductase (GPX/GRD). Some amounts of all the antioxidant enzymes, which may protect spermatozoa from axidative attack, are also made by the epididymis during storage. The glutathione peroxidase/reductase enzymes play a central role in the defense against oxidative damage in human sperm.Results and Discussion: A decrease in levels of reduced glutathione (GSH) during sperm production is known to disrupt the membrane integrity of spermatozoa due increased oxidative stress. Intracellular glutathione levels of spermatozoa are known to be decreased in certain populations of infertile men.Conclusion: Glutathione is not only vital to sperm antioxidant defenses, but selenium and glutathione are essential to the formation of “ phosholipid hydroperoxide glutathione peroxidase” -an enzyme present in spermatids - which becomes a structural protein in the mid-piece of mature spermatozoa. When either substance is deficient, it can lead to instability of the mid-piece of the spermatozoa, resulting in defective motility. Free radical scavengers - such as glutathione-that restore the structure and function of poly - unsaturated fatty acids (PUFA) in the cell membrane, can be used to treat these cases.

Objective: Contraception had been a matter of concept in modern medicine from the sixties when World Health Organization asked governments to make coalitions with to search and find safe and reliable contraception methods. But in Iranian Traditional Medicine (TIM), contraception had been used more than thousand years ago. The ancients know about different contraception methods both for men and women, for short time or lifelong. They believed that contraception methods should be kept as a secret and it could be used only in special situation; when pregnancy was a danger for woman.Materials and Methods: Here we do a search in the most important TIM manuscripts and classified the different contraception methods as following. Besides we examined the effect of decoction, boiled and different fractions of Ruta graveolens L. on human sperm characteristics in vitro.Results: According to our findings, usage of this plant as a male contraceptive is unique to TIM. The results showed that their choice was a right one because coumarins of this plant in addition to its hyperosmolarity and acidic pH could immediately immobilize the spermatozoa and arrest the acrosomal reaction. Meanwhile there was no adverse effect on viability, mitochondrial function or chromatin structure. Classification of contraception methods in TIM: 1- Changing intercourse position in order to diminish the semen arrival to female genitalia. 2- Rapid removement of semen from female genitalia. 3- Mechanical barriers 4- Vaginal suppositories with different materials 5- Topical application of remedies on penis called “Tela” 6- Oral contraceptive for both men and women 7- Usage of “Talyghat”.Conclusion: The results showed that TIM used a wide range of methods for contraception which some were so ingenious. The experimental findings support the TIM unique idea about usage of Ruta graveolens L. as contraception and it reveals that knowing more about traditional remedies and performing more trials, may let to find new ways for contraception in future.

Objective: Allium cepa (onion) has a good effect on diseases treatment worldwide and have been used since ancient times as a medicinal and food origin. Recently several reports have shown that onion has high antioxidant activity. As antioxidants have essential effect on sperm health parameters, we investigated the effect of onion bulbs fresh juice in Spermatogenesis cycle in rats.Materials and Methods: Wistar male rat (n=30) were allocated into three groups, control (n=10) and two test groups (each of ten). Animals in test groups were subdivided into groups of 2 that received fresh onion juice equivalent to 0.5 & 1g/Rat/day fresh onion. Fresh onion juice was administered with gavages for 20 consecutive days. Animals were kept in standard condition. On twentieth day, the testes of rats in the all groups were removed and sperm was collected from epididymis and was prepared for analysis.Results: Serum total testosterone significantly increased in whole test groups (p<0.05) and level of LH significantly increased only in the group that received the high dose of fresh onion juice, (p<0.05), but the level of FSH did not differ between experimental and control groups. The percentage of sperm viability and motility in both test groups significantly increased (p<0.05), but the sperm concentration significantly increased only in the group that received the high dose of freshly extracted onion juice, (p<0.05). It was evident that there was no difference on sperm morphology and testis weight in test groups comparing to control group.Conclusion: In our study freshly prepared onion juice has significantly affected the sperm number and percentage of viability and motility; it seems that using 4g/kg of freshly prepared onion juice is effective in sperm health parameters.