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This study has documented the major types of lineage progenitor cells at the second level of cell differentiation after the establishment of the primary germ layers in ectopic human embryos in vivo. These correspond to stages 8 and 9 of embryogenesis (weeks 3-4) in the Carnegie collection. The aim of this study was to provide images of fine structure of tissue progenitor cells to compare them with current imaging of their equivalent stem cells identified using fluorescent stem cell markers. These include neural, mesenchymal, endodermal, ectodermal (epidermal) and haematopoietic progenitor cells, including those for amniotic, yolk sac and chorionic tissues that are used in current stem cell research. Neural induction by the notochord has been imaged. This study should give valuable clues to understand the pattern of cell differentiation of embryonic stem cells (ESC) in vitro, which are more or less mimicked in ESC colonies, embryoid bodies and neurospheres as documented in the literature. The fine structure of week-3 and week-4 human ectopic embryos is presented to demonstrate progenitor tissue cells that will eventually form the brain, spinal cord, skin, gut, heart, blood, muscle, bone and other tissues of the human body later on in development. These images should help stem cell researchers using fluorescent markers and other techniques to identify embryonic and adult stem cells in culture.
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Células-Tronco/citologia , Diferenciação Celular , Desenvolvimento Embrionário , Células-Tronco Embrionárias/citologia , Endoderma/patologia , Epiderme/patologia , Feminino , Corantes Fluorescentes/farmacologia , Camadas Germinativas/patologia , Células-Tronco Hematopoéticas/citologia , Humanos , Mesoderma/patologia , Microscopia Eletrônica de Transmissão/métodos , Notocorda/patologia , Gravidez , Gravidez Ectópica , Somitos/patologia , Distribuição Tecidual , Saco Vitelino/citologiaRESUMO
The fine structure of the three germ layers in human ectopic embryos (stage 7) have been documented by digital light and electron microscopy. The formation of ectoderm, endoderm and mesoderm and notochordal cells, and also the extraembryonic membranes, amnion and yolk sac, are imaged. The germ layers give rise to all the cells and tissues of the human body. Possible clues to the early differentiation of embryonic stem cells (ESC) in vitro were obtained, since these events are more or less mimicked in cultures of ESC derived from the inner cell mass of human blastocysts. The findings are discussed with reference to previous studies on the fine structure of ESC using the same technique.
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Técnicas de Cultura Embrionária , Células-Tronco Embrionárias/citologia , Camadas Germinativas , Âmnio/fisiologia , Diferenciação Celular , Ectoderma/patologia , Endoderma/patologia , Humanos , Mesoderma/patologia , Microscopia Eletrônica/métodos , Microscopia Eletrônica de Transmissão/métodos , Mitose , Linha Primitiva/patologiaRESUMO
Mullerian anomalies which cause infertility in women were described by different classification systems. We report a rare case of uterine anomaly in a 16-year-old patient presented with primary amenorrhea. Her diagnostic laparoscopy findings revealed two uterine rudimentary horns on either side of the upper pelvis with a hypoplastic noncavitated central uterus. The pathogenesis of this anomaly may not be clearly defined but it was stated that these occur due to the developmental defects in embryo. This case report is one of the rarest cases presented and may signify the Mullerian duct anomaly.
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Sirenomelia is a very rare developmental abnormality which is characterized by fusion of lower limb to form a single limb. This condition is often associated with internal organ abnormality and is considered incompatible with life. Sirenomelia is sporadically reported from across the world, but no case associated with artificial reproductive technology (ART) has not been reported. We report a case of sirenomelia in a 29-year old woman who conceived by ART, which to our knowledge is the first reported case in ART. The defect was detected early during first trimester and the pregnancy was terminated.
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A 31-year-old woman, who is a known case of polycystic ovary syndrome, underwent modified radical hysterectomy and right ovarian transposition to the anterior abdominal wall for endometrioid adenocarcinoma Grade II. She then visited our facility for in vitro fertilization (IVF) and surrogacy in the year 2016. Three cycles of IVF were performed using gonadotropin-releasing hormone (GnRH) antagonist in the first two attempts and GnRH agonist in the third attempt, with percutaneous technique of oocyte retrieval from the transpositioned right ovary. In the third attempt in July 2017, we were able to retrieve five oocytes and subsequently froze three embryos and one blastocyst. The surrogate underwent sequential transfer in June 2018 which resulted in a positive clinical singleton pregnancy. The outcome of IVF following percutaneous oocyte retrieval led to a successful pregnancy and subsequent delivery by emergency cesarean by a healthy female baby on February 16, 2019. This is the first reported case of ovarian hyperstimulation and percutaneous aspiration of oocytes from a transpositioned right ovary (subcutaneous plane) in our country. Global literature survey revealed publications only of transabdominal retrieval of ovaries moved above the pelvic brim.
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An open, prospective, multicenter study was designed to assess the efficacy and safety of dydrogesterone in the post-laparoscopic treatment of endometriosis in Indian patients. Ninety-eight patients suffering from minimal, mild, moderate or severe endometriosis, with or without infertility, who had undergone laparoscopy, were treated with dydrogesterone 10 mg/day (or 20 mg/day in severe cases) orally from day 5 to day 25 of each cycle for 3-6 months. Pelvic pain, dysmenorrhea and dyspareunia improved significantly (p < 0.05) after the first cycle of treatment. By the end of the sixth cycle, the reduction in pelvic pain, dysmenorrhea and dyspareunia was 95%, 87% and 85%, respectively. The amount of menstrual bleeding fell significantly (p < 0.05) after 2 months (-12%) and this significant reduction was maintained until the end of the study. The duration of bleeding was also reduced significantly (p < 0.05) throughout the study, starting after the first month of treatment (-10%). A total of 21.1% of the patients were considered cured and 66.7% showed improvement. Overall, dydrogesterone therapy was rated as excellent to good by 74% of patients and 70% of physicians. No adverse events were reported. In conclusion, dydrogesterone is an effective and safe post-laparoscopic treatment for endometriosis.
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Didrogesterona/uso terapêutico , Endometriose/tratamento farmacológico , Endometriose/cirurgia , Progestinas/uso terapêutico , Adulto , Terapia Combinada , Dismenorreia/tratamento farmacológico , Dispareunia/tratamento farmacológico , Feminino , Humanos , Laparoscopia , Pessoa de Meia-Idade , Satisfação do Paciente , Dor Pélvica/tratamento farmacológico , Estudos ProspectivosRESUMO
The fine structure of human oogonia and growing oocytes has been reviewed in fetal and adult ovaries. Preovulatory maturation and the ultrastructure of stimulated oocytes from the germinal vesicle (GV) stage to metaphase II (MII) stage are also documented. Oogonia have large nuclei, scanty cytoplasm with complex mitochondria. During folliculogenesis, follicle cell processes establish desmosomes and deep gap junctions at the surface of growing oocytes, which are retracted during the final stages of maturation. The zona pellucida is secreted in secondary follicles. Growing oocytes have mitochondria, Golgi, rough endoplasmic reticulum (RER), ribosomes, lysosomes, and lipofuscin bodies, often associated with Balbiani bodies and have nuclei with reticulated nucleoli. Oocytes from antral follicles show numerous surface microvilli and cortical granules (CGs) separated from the oolemma by a band of microfilaments. The CGs are evidently secreted by Golgi membranes. The GV oocytes have peripheral Golgi complexes associated with a single layer of CGs close to the oolemma. They have many lysosomes, and nuclei with dense compact nucleoli. GV breakdown occurs by disorganization of the nuclear envelope and the oocyte enters a transient metaphase I followed by MII, when it is arrested and ovulated. Maturation of oocytes in vitro follows the same pattern of meiosis seen in preovulatory oocytes. The general organization of the human oocyte conforms to that of most other mammals but has some unique features. The MII oocyte has the basic cellular organelles such as mitochondria, smooth endoplasmic reticulum, microfilaments, and microtubules, while Golgi, RER, lysosomes, multivesicular, residual and lipofuscin bodies are very rare. It neither has yolk nor lipid inclusions. Its surface has few microvilli, and 1-3 layers of CGs, aligned beneath the oolemma. Special reference has been made to the reduction and inactivation of the maternal centrosome during oogenesis. The MII spindle, often oriented perpendicular to the oocyte surface, is barrel-shaped, anastral and lacks centrioles. Osmiophilic centrosomes are not demonstrable in human eggs, since the maternal centrosome is nonfunctional. However, oogonia and growing oocytes have typical centrioles, similar to those of somatic cells. The sperm centrosome activates the egg and organizes the sperm aster and mitotic spindles of the embryo, after fertilization.
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Centrossomo/fisiologia , Oócitos , Oogônios , Feminino , Humanos , Oócitos/fisiologia , Oócitos/ultraestrutura , Oogônios/citologia , Oogônios/ultraestrutura , Organelas/ultraestrutura , Folículo Ovariano/citologia , Folículo Ovariano/ultraestrutura , Ovário/citologia , Ovário/ultraestrutura , Óvulo/citologia , Óvulo/ultraestruturaRESUMO
We report the first documented successful birth of twins following preimplantation genetic screening (PGS) of cleavage stage embryos by array comparative genomic hybridization (CGH) technology, in South India. The case was a 28-year-old woman with the previous history of preclinical pregnancy and a miscarriage in two attempted in vitro fertilization cycles. Day 3 cleavage stage embryos were generated by conventional long protocol with the use of a gonadotropin-releasing hormone analog and a combination of recombinant folliculotropins and human menopausal gonadotropins. Intracytoplasmic sperm injection of oocytes thus obtained was performed, and 10 selected embryos underwent PGS using the array CGH technique. Two normal blastocysts were transferred to the patient, and she conceived twins. She delivered at 35 weeks of gestation by elective cesarean on November 19, 2014. She delivered a healthy male and female baby weighing 2.19 kg and 2.26 kg, respectively. Postnatal evaluation of babies was also normal, and the hospital course was uneventful. PGS has a definitive indication in assisted reproductive technology programs and can be utilized to improve pregnancy rates significantly.
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BACKGROUND AND OBJECTIVE: Male infertility has been associated with aneuploidies and structural chromosomal abnormalities, Yq microdeletions and specific gene mutations and/or polymorphisms. Besides genetic factors, any block in sperm delivery, endocrine disorders, testicular tumours, infectious diseases, medications, lifestyle factors and environmental toxins can also play a causative role. This study aimed to determine the constitutional karyotype in infertile males having normal female partners in a south Indian population. MATERIALS AND METHODS: A total of 180 men with a complaint of primary infertility ranging from 1 to 25 years were screened for chromosomal abnormalities through conventional analysis of GTG-banded metaphases from cultured lymphocytes. RESULTS: Four individuals were diagnosed to have Klinefelter syndrome. Two cases exhibited reciprocal translocations and one showed a maternally inherited insertion. Polymorphisms were seen in sixty-seven patients (37.2%). CONCLUSION: The occurrence of chromosomal abnormalities in 4.6% and variants involving the heterochromatic regions of Y, chromosome 9 and the acrocentric chromosomes in 38.2% of the infertile men with an abnormal seminogram strongly reiterates the inclusion of routine cytogenetic testing and counselling in the diagnostic work-up prior to the use of assisted reproduction technologies.
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OBJECTIVE: To report a case of successful pregnancy in a patient with 46,XY karyotype with primary ovarian failure. DESIGN: Case report. SETTING: Fertility Research Center, G.G. Hospital, Chennai, Tamil Nadu, India. PATIENT(S): A 27-year-old woman with hypoplastic uterus, normal fallopian tubes on both sides, and gonadal dysgenesis. INTERVENTION(S): Chromosomal analysis, diagnostic laparoscopy, donor oocyte program, gamete intrafallopian transfer, and gonadectomy. MAIN OUTCOME MEASURE(S): Response to hormone replacement therapy and the probability of achieving a pregnancy by a tubal procedure. RESULT(S): Treatment was successful, and the patient delivered a live baby. CONCLUSION(S): A hypoplastic uterus of patients with the 46,XY karyotype can be stimulated by the use of cyclical steroid therapy to accommodate pregnancy and facilitate tubal procedures in patients with normal fallopian tubes.
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Transferência Intrafalopiana de Gameta , Disgenesia Gonadal 46 XY/complicações , Infertilidade Feminina/terapia , Adulto , Estrogênios Conjugados (USP)/uso terapêutico , Feminino , Humanos , Infertilidade Feminina/etiologia , Gravidez , Resultado da GravidezRESUMO
AIM: The aim of the following study is to compare pregnancy rates between the use of non-temperature and temperature controlled centrifugation on semen preparation technique in intrauterine insemination. MATERIALS AND METHODS: The retrospective study was conducted on 671 patients of idiopathic infertility who underwent homologous artificial insemination at Fertility Research Center from the period of January 2007 to September 2012. The couples were randomized into two groups namely, Group A-patients (n = 303) being treated with sperm prepared by using non-temperature controlled centrifuge and Group B-Patients (n = 368) being treated with sperm prepared by temperature (37°C) controlled centrifuge. RESULTS AND CONCLUSION: The clinical pregnancy rate Group A was 13.86% and Group B was 12.77%. The clinical pregnancy fetal loss rate between the two groups was 38% and 42% respectively. The delivery rate per transfer was 62% and 58% respectively. No statistically significant difference was found between these two groups.
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Heterotropic pregnancy, although a rare condition, is associated with a greater frequency in assisted reproduction. It occurs in approximately 1 in 100 pregnancies conceived by in vitro fertilization (IVF) particularly when multiple embryos are transferred into the uterus. We report a case of heterotropic pregnancy following IVF with the rupture of an isthmo-cornual pregnancy at 12 weeks of gestation with uneventful progression of the intrauterine pregnancy. Laparotomy was performed for the excision of the isthmo-cornual pregnancy. The intrauterine pregnancy continued uneventfully. A female baby was delivered by elective cesarean section at 33 weeks.
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Several genetic factors have been found to be associated with recurrent pregnancy loss (RPL). However, not many attempts have been made to associate the mitochondrial DNA (mtDNA) variations with RPL. Therefore, we have analyzed the complete mtDNA of 100 women with RPL and 12 aborted fetal tissues. Our analysis revealed a total of 681 variations, most of which were in NADH Dehydrogenase (ND) genes that encode mitochondrial enzyme Complex I. Presence of T4216C variation (ND1 gene) in 9% of the RPL women and several pathogenic, and novel mutations suggest the role of mtDNA variations in RPL.
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Aborto Habitual/genética , DNA Mitocondrial/genética , Polimorfismo Genético , Adulto , Complexo I de Transporte de Elétrons/genética , Feminino , Frequência do Gene , Humanos , Índia , Proteínas Mitocondriais/genética , GravidezRESUMO
OBJECTIVE: To evaluate the reproductive outcome following hysteroscopic septum resection in patients with primary and secondary (recurrent pregnancy loss [RPL] and bad obstetric history [BOH]) infertility. STUDY DESIGN: Retrospective study. MATERIALS AND METHODS: Hysteroscopic septum resection was performed on 26 patients with a history of either recurrent pregnancy loss, BOH or infertility. The septum resection was performed using a bipolar versapoint system. Reproductive performance of these patients after septum resection was analyzed. The main outcome measures were clinical pregnancy and live birth rates. RESULTS: Hysteroscopic septum resection was performed on seven patients with the history of secondary infertility. Post operatively, the pregnancy rate was 86% (n=6), and the live birth rate was 67% (n=4). After septum resection in 19 primary infertile patients, 6 (32%) patients conceived which resulted in live birth rates of 67% (n=4). CONCLUSION: Hysteroscopic septum resection using bipolar versapoint system is an effective and safe approach for the removal of septum. Hysteroscopic septum resection in women with septate uterus significantly improves the live birth rates and future fertility is not impaired.
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We report the first twin pregnancy and birth in India after the transfer of embryos generated from vitrified and thawed oocytes to a 41-year-old woman who was in a donor program. Embryos were generated from the microinjection of pre-prepared sperms into vitrified oocytes. Twin male babies weighing 750 and 860 g were born by emergency cesarean section. Vitrification is one of the most promising options in cryopreservation and preservation of embryos and oocytes.
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Premature ovarian failure (POF), that is, amenorrhea before 40 years of age can be attributed to a variety of etiologies. Approximately 1% of women before 30 years are diagnosed with POF. Spontaneous ovulation leading to pregnancy in POF is even a rarer entity. We report a case where congenital malformations were diagnosed in a fetus following spontaneous ovulation in a case of POF. A 33-year-old woman presented to our center with primary infertility. On complete work up, she was diagnosed with POF and conceived with hormone replacement therapy and donor oocyte program. She delivered a healthy female baby through caesarean section. The patient reviewed later with amenorrhea of 40 days and pregnancy was confirmed. However, during antenatal follow-up congenital anomalies in fetus were diagnosed sonographically. The decision for termination of pregnancy was taken. To conclude, we recommend large-scale retrospective analysis that would define medical guidelines in cases of pregnancy following spontaneous ovulation in POF.
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We report the first pregnancy and birth in India after the transfer of embryos generated from frozen- thawed oocytes. A 29-year-old woman with previous bad obstetric history and an abnormal karyotype, necessitating donor oocyte programme. Embryos were generated by microinjection of frozen-thawed sperms into thawed human oocytes (intracytoplasmic sperm injection). This resulted in an healthy male baby with a birth weight of 2.54 kg which was born by cesarean section at 35-36 weeks of gestation with normal follow-up. Thus oocyte cryopreservation can be performed with reproducible success leading to a viable offspring.