There have been some proposals that stem cells exist in the ovarian surface epithelium (OSE) of the adult human ovary; however, no direct evidence of such cells has been given until now. The aim of ...this study was to isolate the putative ovarian stem cells (OSCs) from the OSE layer in women with no naturally present oocytes and follicles—20 postmenopausal women and five women with premature ovarian failure. Small round cells with a bubble-like structure and diameters from 2 to 4 μm were isolated from the material obtained by OSE scraping. They expressed early embryonic developmental markers such as stage-specific embryonic antigen-4 and Oct-4, Nanog, Sox-2, and c-kit transcription markers, and they displayed prominent c-kit immunohistochemical staining. These cells were separated by density gradient centrifugation and grown
in vitro, where they proliferated. Some of them grew intensively and reached a diameter of approximately 20 μm after 5–7 days. In the OSE cell culture, oocyte-like cells developed, which reached a diameter of up to 95 μm and expressed Oct-4A, Oct-4B, c-kit, VASA, and ZP2 transcription markers, corresponding to early oocytes. They did not express SCP3 meiotic marker. In conclusion, the discovered cells are proposed to represent the adult OSCs with the expression of embryonic stem cell markers. The expression of germ lineage marker c-kit points toward their primordial germ cell ancestry. A new term “embryonic-like stem cells of the adult” is proposed for embryonic-like stem cells that might persist in various tissues and organs of adults. These findings could be used for further studies aimed at the autologous treatment of ovarian infertility and degenerative diseases.
To evaluate the involvement of immune abnormality in patients with idiopathic premature ovarian insufficiency (POI). In addition to the known etiology, autoimmune disorders may be a pathologic ...mechanism for POI.
Our study was a prospective controlled trial. Twenty women with POI, reasons other than autoimmune excluded, were enrolled in this study. The control group consisted of 17 healthy women. In both groups, family and personal history were taken and the levels of follicle stimulating hormone, luteinizing hormone, thyroid-stimulating hormone, prolactin, anti-Müllerian hormone, inhibin B, antithyroglobulin and antithyroid peroxidase antibodies were determined. Antiovarian antibodies and subpopulations of peripheral blood T-lymhocytes were also determined.
Participants in the study group exhibited hypergonadotropichypogonadism, while high levels of follicle stimulating hormone and low levels of inhibin B and anti-Müllerian hormone were observed. In 16 (80%) patients, POI was associated in their personal and familial history with another autoimmune disease. Fifty percent of patients presented highly elevated antithyroid antibodies. The lymphocyte subset, especially B cells, was significantly higher (p=0.014), and peripheral regulatory lymphocytes CD25+ high were significantly lower (p=0.015) in the study group than in the control group. Anti- ovarian antibodies were detected in 20% of patients with POI.
We presume that the presence of anti-ovarian antibodies together with abnormalities of cellular immunity may in some cases potentially represent the involvement of an autoimmune mechanism in idiopathic POI.
A fast and selective analytical method, used to determine the different lysophosphatidic acid (LPA) species in serum, has been developed and validated. LPA species were quantitatively extracted from ...serum using methanol–chloroform (2:1, v/v). The proteins were precipitated by this solvent mixture and separated by centrifugation in one step. LPA levels were determined in clear extracts using the HPLC-MS/MS method. The linearity of this method was established in the concentration range between 0.1 and 16
μM for all LPA species with a correlation coefficient greater than 0.99. Recovery of all LPA species determined by the serum, fortified at approximately 1
μM and 2–3
μM, was between 93% and 111% with an average R.S.D. of less than 8%. This method was used to determine LPA in numerous sera of healthy controls, patients with benign ovarian tumours and ovarian cancer at different stages. Significantly higher total LPA levels were determined in the sera of patients with different types of tumours (benign and malignant).
Summary
High seminal reactive oxygen species (ROS) are related to poor semen quality and impaired fertilization. We aimed at finding whether there is an association between ROS and fertilization, ...embryo quality and pregnancy rates after conventional in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI). In prepared semen of 147 male partners of infertile couples, ROS were assessed with luminol chemiluminescence. Spermiogram was assessed in native semen. ROS were negatively correlated with standard sperm characteristics and testicular volume, and positively with abnormal sperm head morphology. Fertilization rate and embryo morphology on day 2 and on day 4 were assessed in 41 IVF and 106 ICSI cycles. The influence of maternal (female age and number of oocytes) and paternal (sperm motility, morphology and ROS) factors on fertilization and embryo quality were assessed by means of regression analyses. After IVF, fertilization and pregnancy rates were negatively associated with ROS level (p = 0.031 and 0.041, respectively). In case of higher ROS, significantly fewer ICSI‐derived embryos (p = 0.036) reached the morula‐blastocyst stage on day 4. High seminal ROS levels are associated with impaired sperm fertilizing ability and lower pregnancy rates after IVF. In ICSI, a negative association of ROS with embryo development to the blastocyst stage has been observed.
BACKGROUND: Fragile X premutation carriers are at increased risk of premature ovarian failure (POF), which is usually defined as menopause before the age of 40 years. METHODS: We evaluated 83 women ...with sporadic premature ovarian failure, treated at the Department of Obstetrics and Gynaecology, University Medical Centre, Ljubljana, between 1991 and 2001. There was no family history of mental retardation in any of the patients. They were phenotypically normal and had normal female karyotype (46,XX), without a past history of pelvic surgery, chemotherapy or autoimmune diseases. RESULTS: The premutation in the FRAXA locus was found in four of the women screened (4.8%; 95% confidence interval 1.9–11.7). This prevalence (1 in 21) was statistically significantly higher than expected in the female Caucasian population. CONCLUSION: In this study we have confirmed an important association between FRAXA premutation and the pathogenesis of POF. This result has practical implications for genetic counselling and fertility treatment.
Abstract Objective To evaluate the effect of counseling on women's adherence to hormone therapy (HT) in Slovenia following the publication of the results of the Women's Health Initiative prospective ...study. Method In this 24-month prospective, randomized, controlled study with 125 women in early menopause, those in the study group (n = 64) attended oral presentations on menopause and HT whereas those in the control group (n = 63) did not. Data were collected from 2 questionnaires, one completed before starting HT and the other during follow-up visits after 3, 6, 12, and 24 months. The results were analyzed according to the intention-to-treat principle. Results In the second year, an improved quality of life and friends critical of HT negatively affected adherence to HT. After 24 months, 47% of the participants in the study group and 32% in the control group were still using HT. The difference between the 2 groups in the rates of participants who discontinued HT was the same in the first than and in the last 12 months of the study (49 76.6% vs 15 23.4%). Conclusion Since there were no changes in discontinuation rates between the groups from one study period to the other, and a total of 15 women in each group stopped using HT in the second year of the study, the effect of the educational presentations on HT adherence was long-term rather than temporary.
In the present study, differences in leptin levels between different groups of male patients presenting with infertility problems and possible correlations between leptin levels and clinical, ...spermiological, histological and hormonal characteristics were examined. Two hundred and ten male partners from infertile couples were included in the study. Based on clinical examination, spermiogram and testicular histology results, patients were divided into four groups: 42 men with non-obstructive azoospermia, 15 men with obstructive azoospermia, 68 men with oligoasthenoteratozoospermia and 85 men with normozoospermia. Serum levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), inhibin B, testosterone, sex hormone binding globulin (SHBG) and leptin were measured. After adjustment for body mass index, there was a negative correlation between serum levels of leptin and inhibin B, total testosterone and SHBG (r = -0.189, p = 0.009, r = -0.250, p = 0.001 and r =-0.221, p = 0.003 respectively) but there was no correlation between leptin and classical sperm characteristics. Our results therefore demonstrate a link between leptin and testicular function, independently of FSH and LH, possibly involving testosterone and SHBG through a regulation of Leydig cell function.
Problem
Interleukin (IL) IL‐12/IL‐18 are involved in uterine NK cells control of uterine vascular development. Polymorphisms in the IL‐12/IL‐18 genes could modify the cytokine balance, which might ...result in an increased susceptibility to recurrent spontaneous abortion (RSA).
Method of Study
A case–control study was conducted to determine the association between the IL12 (I/D) and IL18 (−607C>A, −137G>C) gene polymorphisms and the risk of RSA in 125 women with RSA and in 136 controls.
Results
The frequencies of DD, ID, II for IL‐12 were, 25.6%, 52.8% and 21.6% respectively, in patients versus 21.3%, 51.5% and 27.2% respectively in controls; the frequencies of CC, CA, AA genotypes for IL‐18 (−607) were, 34.4%, 54.4% and 11.2% respectively in patients versus 30.1%, 58.1% and 11.8% respectively in controls; the frequencies of GG, GC, CC genotypes for IL‐18(−137) were 47.2%, 43.2% and 9.6% respectively in patients and 45.6%, 46.3% and 8.1% respectively in controls.
Conclusion
IL‐12B and IL‐18 promoter gene polymorphisms were not associated with RSA in our women.
BACKGROUND Androgen administration for hormonal replacement therapy (HRT) in postmenopausalwomen has recently been involved in many panels and investigations. The objective forthe introduction of ...androgen replacement in postmenopausal women was the opinionthat impaired libido and well-being in women with normal estrogen concentration (withHRT or in reproductive period) is the consequence of androgen deficiency.Physiology of female androgen production in women emphasizes the production ofdehydroepiandrosteron (DHEA) as crucial precursor of human sex steroid biosynthesistogether with its sulphate ester (DHEAS). DHEA and DHEAS do not have androgen activityunless they are converted to testosterone and dihydrotestosterone, which can both bind and activate androgen receptor. The ability of ovarian theca cells to synthesize androgenspersists also after menopause, the fact being illustrated by finding that bilateral oophorectomy in postmenopausal women leads to a significant decrease in circulating levels ofandrogen. The definition of female androgen deficiency syndrom (FADS) was presentedas Princeton's consensus meeting three criteria: impaired libido and well-being with normal estrogen values (established estrogen replacement) and serum androgen concentrations below or within the lower quartile of the female normal range. The criticism of Princeton's consensus is focused mainly on loose definition of impaired libido. The similar critical estimation followed new diagnostic term »hypoactive sexual desire disorder« (HSDD).The task force of endocrine societies from Canada, USA and Australia has provided recentguidelines for therapeutic use of androgens in women. They recommended againstmaking a diagnosis of androgen deficiency in women because this clinical syndrome hasnot been defined properly. In spite of some data of clinical effectiveness of androgen replacement in selected population of women, there are no adequate indications andevidence of safety in long term administration. They recommend well designed and focused studies of androgen action in the population of women after surgical castration, inwomen with hypopituitarrism and adrenal insufficiency. CONCLUSIONS Scientific studies and clinical experiences have not provided until now the answers to thequestion: »Whom to treat, when, why and for how long should androgens be used for HRTin postmenopausal women?«
The aim of this study was to trigger the expression of genes related to oocytes in putative ovarian stem cells scraped from the ovarian surface epithelium of women with premature ovarian failure and ...cultured in vitro in the presence of follicular fluid, rich in substances for oocyte growth and maturation. Ovarian surface epithelium was scraped and cell cultures were set up by scrapings in five women with nonfunctional ovaries and with no naturally present mature follicles or oocytes. In the presence of donated follicular fluid putative stem cells grew and developed into primitive oocyte-like cells. A detailed single-cell gene expression profiling was performed to elucidate their genetic status in comparison to human embryonic stem cells, oocytes, and somatic fibroblasts. The ovarian cell cultures depleted/converted reproductive hormones from the culture medium. Estradiol alone or together with other substances may be involved in development of these primitive oocyte-like cells. The majority of primitive oocyte-like cells was mononuclear and expressed several genes related to pluripotency and oocytes, including genes related to meiosis, although they did not express some important oocyte-specific genes. Our work reveals the presence of putative stem cells in the ovarian surface epithelium of women with premature ovarian failure.
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