Prvo rojstvo otroka bolnici po zdravljenju raka materničnega vratu stadija IB1 z ohranitvijo rodne funkcije v UKC-LJ

Rak materničnega vratu lahko prizadene ženske v rodnem obdobju. V stadiju IB1 je mogoče pri zdravljenju rakave bolezni ohraniti rodno funkcijo ženske z radikalno trahelektomijo in odstranitvijo medeničnih bezgavk.To je operacija, pri kateri se odstrani maternični vrat in  paracervikalno tkivo (parametriji) v kombinaciji z odstranitvijo medeničnih bezgavk, telo maternice pa se ohrani. Predstavljamo primer 32-letne nulipare z rakom materničnega vratu stadija 1B1. Bolnica je bila zdravljena radikalno z ohranitvijo rodne funkcije in je po zdravljenju rodila donošenega otroka

ELECTRON MICROSCOPE COMPARISON OF ENDOMETRIUM FROM UTERINE SEPTUM AND ENDOMETRIUM FROM THE LATERAL WALL

Background. Septate uterus is an important risk factor for spontaneous abortion and preterm delivery. The role of septate uterus in infertility is still questionable. The mechanism of the adverse effects of a septate uterus is not yet understood. The basic theory proposes that the septum represents a less suitable environment for a developing embryo when compared to the unaffected uterine wall. The aim of our study was to compare the endometrial surface morphology in women with septate uterus. Material and methods. This prospective observational study includes endometrial biopsies that were taken from  women with uterine septum. We have included 30 consecutive women who came for the hysteroscopic resection of the septum. The operation was scheduled at the time of the implantation window; an endometrial biopsy was performed and samples were taken from the septum and from the lateral wall and compared under electron microscope. Pinopode development stage and the number of endometrial glands were the main outcome measures. Results. Comparison of the endometrium from the septum with that of the lateral wall showed no difference in the number of endometrial glands (2.0 endometrial glands seen at 250 × magnification vs. 2.5; ns ), not even in the pinopode stage (17.7 day vs. 18.1 day, ns). Conclusions. To our knowledge, there are three researches that have compared endometrium from the septum to the endometrium from the lateral wall in infertile women and all have differences in observed parameters. Our results cannot support earlier findings. The question of mechanism, how septum influences on pregnancy should be further investigated in the larger sample

AUTOMATED CONSTRUCTION OF 3D STATISTICAL SHAPE MODELS

Automated segmentation of medical images is a difficult task because of the complexity of anatomic structures, inter-patient variability, and imperfect image acquisition. Prior knowledge, in the form of pointbased statistical shape models (point distribution models) of a structure of interest can greatly assist segmentation to robustly find the structure in a patient's image. Point distribution models are obtained through sets of corresponding landmarks lying on surfaces of training structures. The key to the automated construction of a three-dimensional (3D) statistical shape model is the identification of corresponding landmarks on training shapes, which is a challenging task. This paper presents a novel method for automated construction of 3D point distribution models. Corresponding surface points are obtained by two main steps: 1) volumes of interest (VOI), each containing one training structure, are manually defined, a reference structure is manually extracted from one training VOI and its surface is established and represented by a set of (reference) points, 2) reference landmarks are propagated to other training VOIs by transformations that are obtained by hierarchical elastic registration between the reference and each of the remaining training VOIs. We illustrate our approach using computed tomography data of the lumbar vertebra

USE OF HYALURONAN-RICH TRANSFER MEDIUM FOR A SINGLE BLASTOCYST TRANSFER IN VITRO FERTILIZATION PROCEDURE

Background. The best way to avoid undesirable multiple pregnancies following in vitro fertilization procedure (IVF) is to perform elective single embryo transfer, but the procedure might result in a reduction of the pregnancy rates. Aim of our study was to establish whether a single blastocyst transfer using a hyaluronan rich transfer medium results in higher pregnancy rates in comparison to the transfer using a conventional transfer medium. Material and methods. Our prospective randomized study included 107 patients enrolled in the 1st, 2nd and 3rd classical IVF or intracytoplasmic sperm injection (ICSI) treatment attempt. Patients included were under 37 years of age with at least one blastocyst developed in the procedure. In the study group (47 patients) blastocyst transfers using the hyaluronan rich transfer medium were performed and in the control group (60 patients) the conventional medium was used. The pregnancy rates in the study and in the control group were compared. Results. The average pregnancy rate per single blastocyst transfer was 30 %; there were no twin pregnancies. The single blastocyst transfer using hyaluronan resulted in a non-significantly higher pregnancy rate (11 %). A significantly higher pregnancy rate with the use of hyaluronan was found in the subgroup of patients with two or more blastocysts developed in their 2nd and 3rd IVF attempt (p = 0.045). Conslusions. The single blastocyst transfer results in high implantation rates. Hyaluronan significantly contributes to higher implantation rates in a selected subgroup of patients following previous implantation failure and with multiple blastocysts developed

THE OUTCOME OF LAPAROSCOPIC TREATMENT OF ENDOMETRIOSISASSOCIATED INFERTILITY

Background. Many studies have shown significant reduction of fertility in endometriosis. Mechanisms of infertility are still not completely clear. The aim of our study is to obtain the effectiveness of laparoscopic treatment of infertility for different stages of endometriosis. Methods. The effectiveness of laparoscopic treatment of infertility in endometriosis was observed through a questionnaire that was sent to 281 patients (pts), who had laparoscopic treatment at the Department of Obstetrics and Gynecology Ljubljana during the period from 1999 to 2006 due to endometriosis as the only cause of infertility. 53.4 % (150/281) pts, who answered the questionnaire, were divided into group I (minimal and mild endometriosis) and group II (moderate and severe endometriosis). We established the rate of pregnancy (in total, spontaneous and ART) and compared the results between two groups. Relatively small percentage of answered questionnaires was most likely due to the fact that the questionnaires were sent to pts who were operated several years ago and did not respond or have changed their address. Because the study is still ongoing, we present partial results. Results. For 132 pts wanting to get pregnant the overall pregnancy rate was 76.5 % (101/132). The rate of pregnant pts depended on the stage of endometriosis. In group I 86.8 % (79/91) pts became pregnant – 58. 2 % (46/79) spontaneously, 41.8 % (33/79) with ART. In group II 53.6 % (22/41) pts became pregnant – 68.2 % (15/22) spontaneously, 31.8 % (7/22) with ART. The difference between groups was statistically significant (p < 0.05). Conclusions. Our results are comparable to the results found in the literature. Laparoscopic treatment of infertility is effective in all four stages of endometriosis. According to our results laparoscopic treatment has the primary role in the treatment of endometriosis-associated infertilit

NEW CLASSIFICATION OF SPERM MORPHOLOGY BEFORE ICSI AT MAGNIFICATION OF X6000

Background. At conventional ICSI, sperm to be injected into the oocyte is chosen at magnification of 400 times which does not permit abnormalities of fine sperm structures to be seen. By using a new method, intracytoplasmic morphologically selected injection (IMSI), motile sperm with a normal head, base and no vacuoles present in the head can be selected at magnification of 6000 times. Vacuoles in the sperm head reflect the damage of nucleus and/or DNA (fragmented or single-stranded DNA). Spermatozoa with vacuoles in the head have lower fertilization capacity and lead to abnormal embryo development, unsuccessful implantation, or spontaneous abortion. Methods. The aim of this study was to evaluate the percentage of men with abnormal sperm morphology (teratozoospermia) included in the ICSI programme, in which it is possible to find a morphologically normal sperm without vacuoles in the head using IMSI method. In each selected sperm we evaluated the morphology of head (normal = 2 points), base (normal = 1 point) and presence of vacuoles in the head (one small or no vacuoles = 3 points). Totally normal sperm (or with one small vacuole in the head) was allotted 6 points. According to this evaluation, sperm were divided into three classes: class I (6–4 points), class II (3–1 points) and class III (0 points). Sperm of class I and II were suitable for injection into the oocyte, whereas sperm of class III were not. Results. Before ICSI, the prepared semen of 13 patients which had teratozoospermia (alone or in combination with other semen abnormalities) was evaluated by the IMSI method. Sperm of class I was found in 5 men (38 %), and sperm of class II in the remaining men (62 %). Sperm of class III was found in all men. On average, 0.7 sperm of class I and 6.1 sperm of class II per man with teratozoospermia were found. Conclusions. Morphologically normal sperm without vacuoles in the head can be found in less than half of men undergoing the ICSI programme. Further research will show, whether it is possible to improve the clinical results of the ICSI method by the sperm selection with the IMSI method

SPERM SELECTION WITH HYALURONAN AND BLASTOCYST DEVELOPMENT AFTER ICSI

Background. Selection of mature sperm for intracytoplasmic sperm injection (ICSI) has recently been made possible by hyaluronan, a naturally occurring substance found in the cumulus cells. Only fully mature sperm have the receptors on the head that effectively bind to the oocyte cumulus, or, to be more precise, to hyaluronan. The aim of this study was to evaluate the differences in fertilization rates and in embryo development to the blastocyst stage after ICSI performed either with sperm selected by hyaluronan or by standard morphological sperm appearance. Methods. This retrospective study involved 17 couples undergoing ICSI. One half of sibling oocytes were fertilized with sperm selected by hyaluronan (84 oocytes), and the other half with sperm selected on the basis of morphological appearance (89 oocytes). The comparison between the two groups involved fertilization rates and the percentages of arrested embryos, and delayed (morulae) and advanced blastocysts. Blastocysts were classified as early, developed and expanded. Results. In the group with hyaluronan-selected sperm 65 % of oocytes fertilized, and in the group with sperm selected by morphological appearance 70 % of oocytes. The fertilization rates did not statistically differ between the two groups, and neither did the percentages of arrested embryos (47 % vs. 62 %), morulae (11 % vs. 11 %), and blastocysts (42 % vs. 26 %). In the group with hyaluronan-selected sperm the shares of expanded and early blastocysts were significantly greater (P ≤ 0.04 and P ≤ 0.002, respectively) than in the group with sperm selected according to morphological appearance. Conclusions. The use of hyaluronan for sperm selection in ICSI does not affect fertilization rates. Hyaluronan-selected sperm improves subsequent development of embryos. A larger study sample is required for the assessment of hyaluronan-selected sperm on the achievement of pregnancies

EFFECT OF MORPHOLOGY AND BLASTOCYST DEVELOPMENT ON THE OUTCOME OF SINGLE-BLASTOCYST TRANSFER IN THE IN VITRO FERTILIZATION PROGRAMME

Background. Multiple pregnancy can be prevented by an elective single blastocyst transfer. The aim of this prospective study was to evaluate the effect of blastocyst development and morphology on the outcome of single blastocyst transfer. Methods. In this study 157 women with single blastocyst transfer were included; their mean age was 33.6 years, range 20–42. Embryos were cultured to the blastocyst stage and evaluated using the Gardner’s classification. An elective single blastocyst transfer was performed in 83 women although 2 or more blastocysts developed (Group 1) and in 74 with only one developed blastocyst (Group 2). Results. In Group 1 (2 or more blastocysts) the pregnancy rate was a significantly higher than in Group 2 (1 blastocyst only) (39.8 % vs. 23.0 %; P < 0.05). After the transfer of a single expanded blastocyst no statistical difference in the pregnancy rate between the two groups was observed (45.5 % vs. 44.6 %). A significantly higher pregnancy rate was found after the transfer of one expanded blastocyst with a good embryoblast and trophoblast morphology compared to the transfer of a single early blastocyst (45.6 % vs. 7.1 %, P < 0.05). Conclusions. After single blastocyst transfer, the likelihood of achieving a pregnancy is higher in women with 2 or more blastocysts developed. And also the likelihood of achieving a pregnancy is higher with an expanded blastocyst transferred with good embryoblast and trophoblast morphology. The blastocyst development is the main predictor of pregnancy, which may reflect the embryonic genetic status

Ovarian surface epithelium stem cells: oogenesis in vitro?

Background: The dogma that the total number of follicles and oocytes available for reproduction are determined at birth, and that one follicle and one oocyte are recruited from the existing pool to mature in each menstrual cycle still persists. There is an increasing experimental evidence that this might not be true. In the ovarian surface epithelium there might be nondifferentiated stem cells that might differentiate into oocytes also in adult life. In this research we aimed at isolating putative stem cells from the ovarian surface epithelium and at evaluating in vitro oogenesis in the ovarian cell culture in vitro in women without naturally present follicles and oocytes in their ovarian cortex – postmenopausal women and women with premature ovarian failure.Methods: Ovarian surface epithelium was scraped from the ovaries of 20 postmenopausal women and 5 women with premature ovarian failure. We tried to find putative stem cells in ovarian scraping and to confirm them by transcription markers Oct-4, Sox-2, and Nanog, and by surface antigen SSEA-4. Cell culture was set up by scraped cells in DMEM/F-12 medium with phenol red, which shows a weak estrogenic activity. Ovarian cell culture was cultured for 20 days in a CO2-incubator at 37 °C and 5 % CO2. Development of cells in the culture was followed and the presence of oocyte-like cells was evaluated by using different methods – evaluation of morphology, transcription markers, surface antigen markers, oocyte immunohistochemical markers, and flow cytometry after propidium iodide staining.Results: In all postmenopausal women and in 4 out of the 5 women with premature ovarian failure putative stem cells were isolated from the ovarian surface epithelium, which were positive for transcription markers of embryonic stem cells Oct-4, Sox-2, Nanog, and for surface antigen SSEA-4. In all these women ovarian cell culture was successfully established. In all cultures oocyte-like cells developed approximately on day 5 of culture, which were positive for transcription marker Oct-4 and immunohistological germ cell markers c-kit, Oct-4, ZP2, DAZL, and even meiotic marker SCP3. Marker SCP3 was expressed in oocyte-like cells developed in ovarian cell culture of postmenopausal women, but not in patients with premature ovarian failure. Flow cytometry after propidium iodide staining revealed a population of potentially haploid oocyte like cells in postmenopausal women. Stem cells developed also into some somatic cell types, including fibroblasts, neuron-, and mioblast-like cells. In patients with premature ovarian failure oocyte-like cells were developed only at the presence of autologous patient’s serum and mostly at the presence of heterologous follicular fluid serum, whereas they did not develop at the presence of fetal calf serum.Conclusions: First results are promising as they indicate the new insight into the physiology of the adult human ovary. In the ovarian surface epithelium putative stem cells with embryonic character are present, which can develop into the oocyte-like cells and other types of cells in vitro. Further research is required to understand better these new findings.</p