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1.
J Assist Reprod Genet ; 37(6): 1477-1488, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32363564

RESUMO

PURPOSE: Little is known about the role of the superior ovarian nerve (SON) in follicular development during the estrus cycle. The aim of the present study was to analyze the role of neural signals arriving through the SON at the ovaries in the regulation of follicular development and ovarian steroid secretion in diestrus 1 of cyclic rats. METHODS: Cyclic rats were subjected to left, right, or bilateral SON sectioning or to unilateral or bilateral laparotomy at diestrus 1 at 11:00 h. Animals were sacrificed 24 h after surgery. RESULTS: Compared to laparotomized animals, unilateral SON sectioning decreased the number of preovulatory follicles, while bilateral SON sectioning resulted in a decreased number of atretic preantral follicles. An important observation was the presence of invaginations in the follicular wall of large antral and preovulatory follicles in animals with denervation. Furthermore, left SON sectioning increased progesterone levels but decreased testosterone levels, which are effects that were not observed in animals that were subjected to right denervation. CONCLUSIONS: At 11:00 h of diestrus 1, the SON was found to stimulate follicle development, possibly via neural signals, such as noradrenaline and/or vasoactive intestinal peptide, and this stimulation induced the formation of follicle-stimulating hormone receptors. The role of the SON in the regulation of ovarian steroid secretion is asymmetric: the left SON inhibits the regulation of progesterone and stimulates testosterone secretion, and the right nerve does not participate in these processes.


Assuntos
Diestro/fisiologia , Estro/fisiologia , Folículo Ovariano/fisiologia , Ovário/inervação , Animais , Modelos Animais de Doenças , Feminino , Hormônio Foliculoestimulante/farmacologia , Humanos , Laparotomia , Hormônio Luteinizante/farmacologia , Tecido Nervoso/patologia , Tecido Nervoso/cirurgia , Folículo Ovariano/inervação , Folículo Ovariano/cirurgia , Ovário/fisiologia , Ovário/cirurgia , Ovulação/fisiologia , Ratos , Testosterona/farmacologia
2.
Ceska Gynekol ; 85(1): 11-14, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32414279

RESUMO

OBJECTIVE: Description of punction of follicular fluid in a patient after ovarian transposition. DESIGN: Case report. SETTING: Department of Obstetrics and Gynaecology, 1st Faculty of Medicine, Charles University and the General Faculty Hospital, Prague. CASE REPORT: We present a case of IVF treatment in a patient with ovarian transposition undergoing punction of follicular fluid and difficulties during this procedure acording to transabdominal route. CONCLUSION: Transabdominal punction od follicular fluid is possible, but with technical difficulities and smaller amount of obtained oocytes. We recomend to aplicate IVF procedures prior to surgical solution.


Assuntos
Cavidade Abdominal/cirurgia , Fertilização in vitro , Líquido Folicular/química , Procedimentos Cirúrgicos em Ginecologia/métodos , Recuperação de Oócitos/métodos , Folículo Ovariano/cirurgia , Feminino , Humanos , Oócitos , Gravidez
3.
Reprod Biol Endocrinol ; 16(1): 71, 2018 Jul 28.
Artigo em Inglês | MEDLINE | ID: mdl-30055625

RESUMO

BACKGROUND: In vivo studies involving molecular markers of the follicle wall associated with follicular fluid (FF) milieu are crucial for a better understanding of follicle dynamics. The inability to obtain in vivo samples of antral follicle wall (granulosa and theca cells) without jeopardizing ovarian function has restricted advancement in knowledge of folliculogenesis in several species. The purpose of this study in mares was to develop and validate a novel, minimally invasive in vivo technique for simultaneous collection of follicle wall biopsy (FWB) and FF samples, and repeated collection from the same individual, during different stages of antral follicle development. We hypothesized that the in vivo FWB technique provides samples that maintain the normal histological tissue structure of the follicle wall layers, offers sufficient material for various cellular and molecular techniques, and allows simultaneous retrieval of FF. METHODS: In Experiment 1 (ex vivo), each follicle was sampled using two techniques: biopsy forceps and scalpel blade (control). In Experiment 2 (in vivo), FWB and FF samples from 10-, 20-, and 30-mm follicles were repeatedly and simultaneously obtained through transvaginal ultrasound-guided technique. RESULTS: In Experiment 1, the thickness of granulosa, theca interna, and theca externa layers was not influenced (P > 0.05) by the harvesting techniques. In Experiment 2, the overall recovery rates of FWB and FF samples were 97 and 100%, respectively. However, the success rate of obtaining samples with all layers of the follicle wall and clear FF varied according to follicle size. The expression of luteinizing hormone receptor (LHR) was mostly confined in the theca interna layer, with the estradiol-related receptor alpha (ERRα) in the granulosa and theca interna layers. The 30-mm follicle group had greater (P < 0.05) LHR expression in the theca interna and ERRα in the granulosa layer compared to the other groups. The overall expression of LHR and ERRα, and the intrafollicular estradiol were higher (P < 0.05 - P < 0.0001) in the 30-mm follicle group. CONCLUSION: The in vivo technique developed in this study can be repeatedly and simultaneously used to provide sufficient FWB and FF samples for various cellular and molecular studies without jeopardizing the ovarian function, and has the potential to be translated to other species, including humans.


Assuntos
Biópsia/veterinária , Cavalos , Folículo Ovariano/cirurgia , Animais , Biomarcadores/metabolismo , Biópsia/instrumentação , Biópsia/métodos , Feminino , Líquido Folicular/metabolismo , Imuno-Histoquímica , Ovário/patologia , Ovário/fisiopatologia , Ovário/cirurgia
4.
Fertil Steril ; 109(5): 940, 2018 05.
Artigo em Inglês | MEDLINE | ID: mdl-29778392

RESUMO

OBJECTIVE: To demonstrate a unique case of direct ultrasound-guided follicle aspiration at time of a laparotomy in a patient with Mayer-Rokitansky-Küster-Hauser Syndrome. DESIGN: Video presentation. SETTING: Academic fertility center PATIENT(S): A 32-year-old gravida 0 with Mayer-Rokitansky-Küster-Hauser Syndrome and bilateral ovarian masses presented as a fertility preservation consult from gynecologic oncology due to the possibility of bilateral oophorectomy. Due to the appearance and size of the left ovary oncology planned to perform an exploratory laparotomy and left oophorectomy, with possible right oophorectomy. The patient and her partner desired embryo cryopreservation with plans for future use in a gestational carrier. She had previously undergone vaginal dilator therapy, however her ovaries were inaccessible transvaginally due to their cephalad location and small caliber of the vaginal pouch. The plan was made to proceed with controlled ovarian stimulation and concurrent ultrasound-guided follicle aspiration of the right ovary at the time of laparotomy following left oophorectomy. INTERVENTION(S): Ultrasound-guided follicle aspiration in vivo at time of laparotomy. MAIN OUTCOME MEASURE(S): Successful controlled ovarian stimulation, oocyte retrieval and embryo cryopreservation. RESULT(S): The patient underwent a long agonist protocol and received a total of 2,525 units of gonadotropin with a peak estradiol of 3,264 pg/ml. She required a total of 9 days of stimulation. The normal right ovary responded as expected, and the left ovary remained unchanged. Following laparotomy and left oophorectomy, direct application of the transvaginal ultrasound probe was used to aspirate all visible follicles on the right side in vivo. Twenty-four oocytes were retrieved, 15 were mature and 5 blastocysts were cryopreserved. Final pathology of left ovary returned as serous cystadenoma. The right ovary was examined by gynecologic oncology prior to and following retrieval and was thought to be normal and remained in situ. CONCLUSION(S): Although the approach described here is not feasible in most cases, this video demonstrates a unique and successful fertility preservation technique by direct ultrasound-guided follicle aspiration in vivo at the time of laparotomy in a Mayer-Rokitansky-Küster-Hauser Syndrome patient and to our knowledge, is the first description of its kind. This retrieval would have otherwise been limited by lack of access transvaginally and limited visualization transabdominally. This combined approach should be considered in future patients with müllerian anomalies and similar complicating factors necessitating laparotomy.


Assuntos
Transtornos 46, XX do Desenvolvimento Sexual/diagnóstico por imagem , Transtornos 46, XX do Desenvolvimento Sexual/cirurgia , Anormalidades Congênitas/diagnóstico por imagem , Anormalidades Congênitas/cirurgia , Preservação da Fertilidade/métodos , Ductos Paramesonéfricos/anormalidades , Folículo Ovariano/diagnóstico por imagem , Folículo Ovariano/cirurgia , Ultrassonografia de Intervenção/métodos , Transtornos 46, XX do Desenvolvimento Sexual/complicações , Adulto , Biópsia por Agulha/instrumentação , Biópsia por Agulha/métodos , Feminino , Preservação da Fertilidade/instrumentação , Humanos , Laparotomia/instrumentação , Laparotomia/métodos , Ductos Paramesonéfricos/diagnóstico por imagem , Ductos Paramesonéfricos/cirurgia , Indução da Ovulação/instrumentação , Indução da Ovulação/métodos , Fatores de Tempo , Ultrassonografia de Intervenção/instrumentação
5.
J Avian Med Surg ; 32(1): 13-18, 2018 03.
Artigo em Inglês | MEDLINE | ID: mdl-29698073

RESUMO

Assisted reproduction techniques in birds have been developed for zootechnical purposes and have been adapted for use in conservation of wild bird species. To develop a technique for obtaining follicles in live hens, 5 Rhode Island red hens ( Gallus gallus domesticus) were anesthetized, and abdominal ultrasound was performed to confirm the presence of ovarian follicles. A left celiotomy then was performed to obtain follicles in different stages of maturation for in vitro fertilization. The follicles were located by digital exploration, then extracted by isolating each follicle with the index finger of each hand, holding it by the stigma, and then applying slight traction towards the exterior of the coelomic cavity until the follicle separated from the ovary. In total, 18 of 30 (60%) follicles obtained were suitable for in vitro fertilization, but only 3 (16%) were fertilized successfully. All birds recovered from the procedure and remained in good condition postoperatively. Perfecting assisted reproduction technique holds potential benefits for determining sex of embryos by blastomeres sexing, supporting the conservation efforts of avian species, and benefiting research areas, such as genetic and biopharmaceutical research.


Assuntos
Galinhas/cirurgia , Fertilização in vitro/veterinária , Folículo Ovariano/cirurgia , Analgésicos Opioides/administração & dosagem , Animais , Antibacterianos/administração & dosagem , Comportamento Animal , Galinhas/fisiologia , Sedação Profunda/métodos , Sedação Profunda/veterinária , Enrofloxacina/administração & dosagem , Feminino , Fertilização in vitro/métodos , Modelos Animais , Folículo Ovariano/diagnóstico por imagem , Folículo Ovariano/fisiologia , Dor Pós-Operatória/tratamento farmacológico , Dor Pós-Operatória/veterinária , Cuidados Pós-Operatórios/veterinária , Tramadol/administração & dosagem , Ultrassonografia/veterinária
6.
Saudi Med J ; 39(3): 290-295, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29543308

RESUMO

OBJECTIVE: To compare follicular reduction prior to human chorionic gonadotropin (HCG) trigger and coasting in terms of ovarian hyper-stimulation syndrome (OHSS) reduction, pregnancy, and cancellation rates in in vitro fertilization/ intracytoplasmic sperm injection (IVF/ICSI) cycles. METHODS:  This study was designed as a prospective study. The setting was the IVF unit at King Abdulaziz Medical City, Riyadh, Kingdom of Saudi Arabia. A total of 39 patients undergoing IVF/ICSI cycles, who were at risk of OHSS, 20 were put into a coasting group and 19 had follicular reduction instead. This occurred between October 2010 and January 2011. Our main outcome was OHSS reduction. RESULTS: Six (30%) women developed OHSS in the coasting group and 2 (10.5%) women developed OHSS in the follicular group (p-value=0.235). The pregnancy rates in the cycles were similar for both groups: 4/20 (20%) in the coasting group and 3/19 (15.8%) in the follicular group (p-value=1.000). The cancellation rate of the cycles was similar for both groups, 6/20 (30%) in the coasting group and 1/19 (5.3%) in the follicular group (p-value=0.09). The median number of punctured follicles was significantly lower in the follicular group (16 follicles, interquartile range (IQR)=21-12) compared to the coasting group (29 follicles, IQR=37.8-19.8, p-value=0.001). The retrieved, fertilized, and cleaved oocytes, as well as the number of embryos transferred, were similar amongst both groups. CONCLUSION: There was no difference between follicular reduction prior to HCG and coasting, in terms of OHSS reduction, pregnancy, and cancellation rates in both the IVF and ICSI cycles.


Assuntos
Gonadotropina Coriônica/administração & dosagem , Folículo Ovariano/cirurgia , Síndrome de Hiperestimulação Ovariana/prevenção & controle , Indução da Ovulação/efeitos adversos , Adulto , Estradiol/sangue , Feminino , Humanos , Síndrome de Hiperestimulação Ovariana/etiologia , Indução da Ovulação/métodos , Gravidez , Taxa de Gravidez , Estudos Prospectivos , Injeções de Esperma Intracitoplásmicas
7.
Urology ; 116: 176-179, 2018 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-29548867

RESUMO

Hermaphroditism is known as ovotesticular disorder of sex development. A 14-year-old boy was admitted with right acute scrotum. Exploration revealed tunica rupture and hematoma, with no viable tissue. After 1 month, he was admitted again with left hemiscrotal pain. Microscopic examination of the left gonad demonstrated foci of hemorrhagic cysts, primordial follicles, and regions of seminiferous tubules. We preserved a testicular tissue and the ovarian part was extracted completely. Long-term follow-up with his hormonal profile is reported. This is a case of ovotesticular disorder presented with acute scrotum and we also tried to reduce long-term hormone therapy, with preservation of testicular part.


Assuntos
Dor Aguda/diagnóstico , Folículo Ovariano/cirurgia , Transtornos Ovotesticulares do Desenvolvimento Sexual/diagnóstico , Escroto/fisiopatologia , Dor Aguda/etiologia , Adolescente , Diagnóstico Diferencial , Feminino , Humanos , Masculino , Folículo Ovariano/anormalidades , Transtornos Ovotesticulares do Desenvolvimento Sexual/complicações , Transtornos Ovotesticulares do Desenvolvimento Sexual/cirurgia , Escroto/cirurgia , Túbulos Seminíferos/anormalidades
8.
Reproduction ; 155(2): 173-181, 2018 02.
Artigo em Inglês | MEDLINE | ID: mdl-29162649

RESUMO

Recently, the influence of adrenergic activity over ovarian function, and thus fertility, has begun to gain importance. Previous studies have shown that adrenergic activity through norepinephrine (NE) participates in the control of follicular development and steroidal secretion from the ovary, among other functions. To examine this phenomenon, the denervation of the gonad has been widely used to observe changes in the ovary's performance. Nevertheless, the effect of the absence of adrenergic nerves in the ovary has only been studied in short times periods. In the present work, we used guanethidine (a drug that produces an irreversible sympathectomy) during the infantile period of rats, and we observed its effects in the adult rat (6 months old). Our results indicate that ovarian NE content is recovered at 6 months old, alongside with an increase of the adrenal content of NE and a dysfunctional celiac ganglion. Together, these results suggest that the recovery of ovarian NE does not come from a neural origin. In addition, ovarian performance was impaired because the changes in follicular development and steroidal secretion are not recovered despite the recovery of ovarian NE content. In conclusion, these results suggest that the nerve-ovarian connections, which are established during infantile development, are necessary for the accurate response of the ovary to sympathetic stimulation.


Assuntos
Estradiol/metabolismo , Norepinefrina/metabolismo , Folículo Ovariano/citologia , Simpatectomia , Sistema Nervoso Simpático/cirurgia , Animais , Feminino , Guanetidina/farmacologia , Folículo Ovariano/efeitos dos fármacos , Folículo Ovariano/cirurgia , Ratos , Ratos Sprague-Dawley , Simpatolíticos/farmacologia
9.
Reprod Biomed Online ; 35(1): 37-41, 2017 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-28483339

RESUMO

In contrast to multifollicular IVF, follicular flushing seems to increase the efficacy of monofollicular IVF treatments such as natural cycle IVF (NC-IVF). However, because follicular flushing causes loss of granulosa cells, it might negatively affect luteal phase length and endocrine function of the luteal body. A prospective cohort Phase II study was performed in 24 women undergoing NC-IVF. Women underwent a reference cycle with human chorionic gonadotrophin-induced ovulation without follicle aspiration and analysis of the length of the luteal phase and luteal concentrations of progesterone and oestradiol. In addition, they underwent a NC-IVF cycle which was performed identically but follicles were aspirated and flushed three times. The luteal phase was shorter in 29.2%, equal in 16.7% and longer in 50.0% of cases following flushing of the follicles. Overall, neither difference in luteal phase length was significant [median duration (interquartile range) in reference cycle: 13 (12; 14.5), IVF (flushing) cycle: 14 (12.5; 14.5), median difference (95% CI): 0.5 (-0.5 to 1.5)] nor median progesterone and oestradiol concentrations. In conclusion, follicular flushing in NC-IVF affects neither the length of the luteal phase nor the luteal phase concentrations of progesterone and oestradiol, questioning the need for luteal phase supplementation.


Assuntos
Estradiol/sangue , Fase Luteal/metabolismo , Recuperação de Oócitos/métodos , Folículo Ovariano/cirurgia , Progesterona/sangue , Adulto , Feminino , Fertilização in vitro/métodos , Humanos , Fase Luteal/fisiologia , Fatores de Tempo
10.
Cell Death Dis ; 8(5): e2817, 2017 05 25.
Artigo em Inglês | MEDLINE | ID: mdl-28542147

RESUMO

In mammalian ovaries, primordial follicles remain in a quiescent state until activation by the surrounding microenvironment. Ovarian intervention, for example, ovarian cystectomy, ovarian wedge resection or laser drilling therapies for polycystic ovarian syndrome, has long been reported to change follicular development by an unknown mechanism(s). Herein, we established a murine model with partial ovarian resection of one ovary unilaterally, with the contralateral ovary undamaged. We found the injury accelerated follicular activation and development through the mTORC1 signaling pathway. Moreover, the stimulation of primordial follicles was restricted near the incision site where the mTORC1 pathway showed sequential activation beginning at the interstitial cells and proceeding to the primordial follicles. Total and polysome-associated RNA-seq revealed the increase of the nerve growth factor (NGF) family member, in both two fractions and immunostaining showed the restricted induction of NGF near the incision site. In cultured newborn ovaries, NGF demonstrated increase of follicular activation, and moreover, the NGF inhibitor K252a effectively blocked activation of primordial follicles stimulated by the surgery. We liken ovulation in mammals to minor tissue trauma, which happens naturally and cyclically in the body. As the increase in NGF accompanied the accumulation of activated primordial follicles after ovulation, our study may represent a common mechanism for selective follicular activation induced by a localized increase in NGF in interstitial cells and mediated via the mTOR signaling pathway. In addition, the NGF inhibitor K252a and the mTOR inhibitor rapamycin constitute good candidates for protecting follicular reserve against over exhaustion after ovarian surgery.


Assuntos
Fator de Crescimento Neural/metabolismo , Folículo Ovariano/citologia , Folículo Ovariano/metabolismo , Transdução de Sinais , Serina-Treonina Quinases TOR/metabolismo , Animais , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Alvo Mecanístico do Complexo 1 de Rapamicina/metabolismo , Camundongos , Folículo Ovariano/cirurgia , Ovulação/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Sirolimo/farmacologia , Células Estromais/efeitos dos fármacos , Células Estromais/metabolismo , Serina-Treonina Quinases TOR/antagonistas & inibidores , Transcrição Gênica/efeitos dos fármacos
11.
J Obstet Gynaecol Res ; 41(2): 264-8, 2015 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-25345318

RESUMO

AIM: We planned to histologically demonstrate unintentional ovarian harm caused by excision of endometriomas. METHODS: One hundred and seven patients with 135 endometriomas had laparoscopic excision of endometriomas at a tertiary health-care clinic over a 6-year period. Records were obtained retrospectively and pathological specimens were reevaluated to demonstrate follicular loss. Tissue damage was scored morphologically and compared according to surgeons' experience and endometrioma size. RESULTS: Ovarian follicles were detected in 80% of endometrioma specimens near the cyst wall. Morphological scores of ovarian follicles were not affected by the surgeon's experience or endometrioma size. CONCLUSION: Surgical excision of endometriomas causes inevitable ovarian follicular loss. As observed in our study, this loss rate could not be traced to surgical experience or ovarian size.


Assuntos
Competência Clínica , Endometriose/cirurgia , Doenças Ovarianas/cirurgia , Folículo Ovariano/cirurgia , Adulto , Endometriose/patologia , Feminino , Humanos , Laparoscopia , Doenças Ovarianas/patologia , Estudos Retrospectivos , Adulto Jovem
12.
Reprod Fertil Dev ; 26(4): 521-6, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-23595420

RESUMO

Two experiments were performed in wood bison during the anovulatory season to establish an effective protocol for ovarian synchronisation. In an untreated control phase, bison cows (n=19) were examined daily to establish the interval to new follicular wave emergence (4.9±0.7 days) for the purposes of comparison with the experimental treatments. In Experiment 1, bison were treated by transvaginal ultrasound-guided follicular ablation (n=9) or with 2mg, i.m., 17ß-oestradiol (n=10). In Experiment 2, bison were treated by follicular ablation (n=9) or with 2mg, i.m., 17ß-oestradiol +100mg, i.m., progesterone (n=10). In Experiment 1, the interval to new wave emergence for control, follicular ablation and 17ß-oestradiol-treated groups was 4.9±0.7, 1.1±0.1 and 3.1±0.4 days, respectively (P<0.05). The degree of synchrony was 2.4±0.4, 0.2±0.1 and 0.8±0.2 days, respectively (P<0.05). In Experiment 2, the interval to new wave emergence for control, follicular ablation and 17ß-oestradiol + progesterone-treated groups was 4.9±0.7, 1.2±0.2 and 3.3±0.3 days, respectively (P<0.05), and the degree of synchrony was 2.4±0.4, 0.2±0.1, and 0.8±0.2 days, respectively (P<0.05). The degree of synchrony did not differ between ablation and hormone treatment groups in either experiment, but was greater in treatment groups than in the untreated control phase. Both follicular ablation and hormone treatment shortened and decreased the variability in the interval to follicular wave emergence in bison, but wave emergence occurred earlier after follicular ablation.


Assuntos
Técnicas de Ablação/veterinária , Anovulação , Bison/fisiologia , Estradiol/farmacologia , Sincronização do Estro/efeitos dos fármacos , Folículo Ovariano/efeitos dos fármacos , Folículo Ovariano/cirurgia , Indução da Ovulação/veterinária , Ovulação/efeitos dos fármacos , Progesterona/farmacologia , Estações do Ano , Animais , Espécies em Perigo de Extinção , Feminino , Folículo Ovariano/fisiologia , Fatores de Tempo
13.
J Assist Reprod Genet ; 30(10): 1279-88, 2013 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-23934019

RESUMO

PURPOSE: To improve human primordial follicle culture. METHODS: Thin or thick ovarian slices were cultured on alginate scaffolds or in PEG-fibrinogen hydrogels with or without bpV (pic), which prevents the conversion of phosphatidylinositol-trisphosphate (PIP3) to phosphatidylinositol-bisphosphate (PIP2) or 740Y-P which converts PIP2 to PIP3. Follicular growth was evaluated by follicular counts, Ki67 immunohistochemistry, and 17ß-estradiol (E2) levels. RESULTS: BpV (pic) had a destructive effect on cultured follicles. Thawed-uncultured samples had more primordial follicles than samples cultured in basic medium and fewer developing follicles than samples cultured in PEG-fibrinogen hydrogels with 740Y-P. There were more atretic follicles in samples cultured on alginate scaffolds than in PEG-fibrinogen hydrogels, and in samples cultured in PEG-fibrinogen hydrogels with 740Y-P than in PEG-fibrinogen hydrogels with basic medium. Ki67 staining was higher in PEG-fibrinogen hydrogels than on alginate scaffolds. E2 levels were higher in thick than in thin slices. CONCLUSIONS: PEG-fibrinogen hydrogels appear to have an advantage over alginate scaffolds for culturing human primordial follicles. Folliculogenesis is not increased in the presence of substances that enhance PIP3 production or with thin rather than thick sectioning.


Assuntos
Técnicas de Cultura de Órgãos , Folículo Ovariano/citologia , Alicerces Teciduais , Adolescente , Adulto , Técnicas de Cultura de Células , Células Cultivadas , Criança , Pré-Escolar , Criopreservação , Feminino , Humanos , Hidrogéis , Neoplasias/patologia , Folículo Ovariano/cirurgia , PTEN Fosfo-Hidrolase/antagonistas & inibidores , Fosfatidilinositóis/química , Engenharia Tecidual , Compostos de Vanádio , Adulto Jovem
14.
J Assist Reprod Genet ; 30(9): 1227-30, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-23842746

RESUMO

PURPOSE: To collect human oocytes from ovaries removed as part of surgical treatment for endometrial carcinoma, and to induce in vitro maturation of such oocytes to obtain material for research on human ovarian aging. DESIGN: Prospective clinical study. SETTING: University Hospital. PATIENTS: Eight patients aged 35-44 years with a preoperative diagnosis of Stage I endometrial cancer agreed to participate in this project. INTERVENTIONS: Surgically removed ovaries were punctured; oocytes were collected from follicular fluid and matured in vitro. Immunofluorescent detection of microtubules and DNA labeling were performed after in vitro maturation. MAIN OUTCOME MEASURES: Number of oocytes collected and their in vitro maturation stage. RESULTS: In total, 87 oocytes were collected, 11 of which had completed metaphase II. Of the oocytes collected, 75 % were from three patients in their 30s, while the remaining 25 % were from five patients in their 40s. Several stages of oocytes were collected and the detection of microtubule arrangement and chromatin in various stages using fluorescence was possible. CONCLUSION: Material for research on human ovarian aging can be obtained from ovaries removed during surgery for endometrial cancer.


Assuntos
Neoplasias do Endométrio/patologia , Técnicas de Maturação in Vitro de Oócitos , Oócitos/crescimento & desenvolvimento , Folículo Ovariano/crescimento & desenvolvimento , Adulto , Neoplasias do Endométrio/cirurgia , Feminino , Fertilização in vitro , Humanos , Meiose/genética , Folículo Ovariano/cirurgia , Ovariectomia , Estudos Prospectivos
15.
Gynecol Endocrinol ; 29(4): 357-60, 2013 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-23305483

RESUMO

With proper and careful selection of patients, fertility-preserving surgery may be feasible in patients with ovarian malignancies. However, the loss of follicles by oophorectomy and chemotherapy results in decreased ovarian reserve, which consecutively affects reproductive capacity. We evaluated postoperative levels of serum anti-Müllerian hormone (AMH) in women with ovarian malignancies to assess the impact of the fertility-preserving surgery with or without the administration of chemotherapy on ovarian reserve. In 13 patients who underwent the fertility-preserving surgery with (n = 9) or without (n = 4) the administration of chemotherapy, serum AMH levels were measured and compared with serum AMH levels measured in patients undergone cystectomy for benign ovarian tumors as a control. We found that the mean AMH level in the treatment group measured 0.9 ng/mL, which was significantly lower than that measured in the control group (4.70 ± 3.77 ng/mL). The possibility of decreased ovarian reserve occurring in patients with ovarian malignancies following treatment with fertility-preserving surgery with or without the administration of chemotherapy should be considered for fertility planning.


Assuntos
Hormônio Antimülleriano/sangue , Preservação da Fertilidade , Folículo Ovariano/cirurgia , Neoplasias Ovarianas/sangue , Adulto , Biomarcadores/sangue , Feminino , Humanos , Laparoscopia , Neoplasias Ovarianas/tratamento farmacológico , Neoplasias Ovarianas/cirurgia , Ovariectomia
16.
Theriogenology ; 79(5): 735-43, 2013 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-23352704

RESUMO

The aims of this study in mares were to: (1) compare preantral follicle parameters between in vitro Biopsy Pick-Up (BPU) and scalpel blade collection methods and between histological and mechanical isolation processing (experiment 1); (2) histologically evaluate preantral follicles (experiment 2); and (3) compare histological analysis with a previously established mechanical isolation technique using a tissue chopper (experiment 3) for ovarian cortical fragments obtained in vivo using a BPU instrument. In experiment 1, preantral follicles were analyzed (N = 220; 90% primordial and 10% primary). Proportions of primordial and primary follicles did not differ (P > 0.05) between tissue collection (BPU vs. scalpel blade dissection) or processing (mechanical isolation vs. histology) methods. Follicle viability and morphology rates were similar (P > 0.05) between tissue collection methods, but mechanical isolation produced more (P < 0.05) morphologically normal follicles than histology. For experiment 2, preantral follicles (N = 332) were analyzed and primordial and transitional (combined) follicles and oocytes were 36.3 ± 0.3 and 26.1 ± 0.3 µm in diameter, respectively, and primary follicles and oocytes averaged 42.9 ± 1.8 and 31.8 ± 2.1 µm. For experiment 3 (188 preantral follicles), within the same animals, the proportion of primordial versus primary follicles was higher (P < 0.03) for histological analysis (98%) compared to tissue chopper analysis (94%), and number of follicles per mg of tissue was not affected (P > 0.05) by processing methods. In conclusion, most parameters evaluated for preantral follicles were similar between histological and tissue chopper processing techniques; hence, mechanical isolation efficiently dissociated equine preantral follicles from the ovarian cortex. Therefore, the tissue chopper could be used to isolate large numbers of morphologically normal equine preantral follicles for cryopreservation and/or in vitro culture.


Assuntos
Cavalos , Recuperação de Oócitos/veterinária , Animais , Criopreservação , Feminino , Biópsia Guiada por Imagem/métodos , Biópsia Guiada por Imagem/veterinária , Recuperação de Oócitos/métodos , Folículo Ovariano/anatomia & histologia , Folículo Ovariano/patologia , Folículo Ovariano/cirurgia
17.
Zygote ; 21(2): 158-61, 2013 May.
Artigo em Inglês | MEDLINE | ID: mdl-22475413

RESUMO

There is a paucity of efficient cryopreservation protocols for primordial follicles enclosed in the ovarian tissue from non-human primates (NHP), in special New World primates. Our objective was to establish an optimal procedure for the recovery of ovarian biopsies from capuchin monkeys. To this end, we adapted a trap door biopsy method. Follicular density and quality of the biopsies were evaluated and ultrasound analysis was performed before and continuously after surgery to assess ovarian structure. Ovarian tissue biopsies recovered by the trap door technique allowed the successful harvesting of primordial follicles from capuchin monkeys, and no complication was recorded. The female cycle was not affected by surgery and no adherence was found thereafter. In conclusion, the adaptation of a trap door biopsy method is a safe procedure and allows recovery of healthy primordial follicles.


Assuntos
Folículo Ovariano/cirurgia , Procedimentos de Cirurgia Plástica/métodos , Animais , Biópsia , Cebus , Feminino , Humanos , Microscopia , Folículo Ovariano/citologia , Folículo Ovariano/diagnóstico por imagem , Ultrassonografia Doppler Dupla
18.
Theriogenology ; 79(2): 267-73, 2013 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-23174772

RESUMO

Ultrasound-guided transvaginal follicle aspiration is used to recover cumulus-oocyte complexes (for IVF) and to synchronize follicular wave emergence (ablation of dominant follicle). Although aspirated follicles are generally supposed to undergo immediate atresia, there are indications that they may remain active. The objective was to evaluate the occurrence and characteristics of residual follicles (RF) after transvaginal follicle aspiration in cattle. Ovarian follicular wave emergence was synchronized in Holstein cows (N = 13) in the presence (groups 1 and 3) or absence (groups 2 and 4) of norgestomet implants. The largest follicle was aspirated at a diameter of 8 mm (groups 1 and 2) or 12 mm (groups 3 and 4). Ovarian follicles were visualized (transrectal ultrasonography) every 12 h after wave emergence. Follicular fluid samples were collected from the largest follicle and from the ensuing RF and concentrations of estradiol and progesterone were determined. After aspiration, 73.2% (52/71) of the follicles refilled with fluid, and a new antrum was detected 12 to 24 h later. Norgestomet did not affect (P > 0.05) RF occurrence or diameter, but in RF from group 4, concentrations of estradiol decreased (-530.7 ± 133.9 ng/mL; P < 0.01) whereas progesterone increased (+429.6 ± 171.7 ng/mL; P < 0.05) relative to preaspiration. In RF, there were three steroidogenesis patterns: (1) high estradiol concentration and high estradiol:progesterone ratio (estradiol-active RF); (2) low estradiol, but high progesterone concentrations (luteinized RF); and (3) low estradiol and low progesterone concentrations (inactive RF). Estradiol-active RF were more likely (P < 0.05) from follicles with high estradiol concentrations (regardless of diameter). In conclusion, fluid-filled structures (RF) with variable steroid production patterns are frequently formed after ultrasound-guided follicle aspiration. The occurrence and features of these RF depended on the diameter and status of these follicles before aspiration.


Assuntos
Bovinos , Folículo Ovariano/fisiologia , Coleta de Tecidos e Órgãos/veterinária , Ultrassonografia/veterinária , Animais , Bovinos/anatomia & histologia , Bovinos/fisiologia , Células do Cúmulo , Estradiol/análise , Feminino , Líquido Folicular/química , Oócitos , Folículo Ovariano/diagnóstico por imagem , Folículo Ovariano/cirurgia , Pregnenodionas/administração & dosagem , Progesterona/análise , Sucção/veterinária , Coleta de Tecidos e Órgãos/métodos
19.
Fertil Steril ; 96(2): 374-8, 2011 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-21703608

RESUMO

OBJECTIVE: To identify a possible marker of follicular depletion in relation to some histologic parameters of endometriotic cysts. DESIGN: Prospective study. SETTING: Università Cattolica del Sacro Cuore, Operative Division of Endocrinological Gynecology. PATIENT(S): Seventy-seven patients (aged 20-40 years) with endometrioma. INTERVENTION(S): Patients underwent laparoscopic surgery for ovarian endometriosis. MAIN OUTCOME MEASURE(S): After excision of the cyst wall, involuntarily removed follicles were correlated with age at surgery and with intrinsic histologic parameters of the specimen (thickness and composition of capsule; size of cyst). RESULT(S): There was a statistically significant relationship between patient age and number of follicles in the histologic section, a statistically significant inverse relationship between size of cyst and number of follicles, and no significant correlation between thickness of the capsule and number of follicles. Fibroblastic-type capsule, most frequently found in younger patients, was associated with removal of a significantly higher number of follicles. CONCLUSION(S): Our study suggests that patient age and cyst dimension are related to the histologic composition of the capsule, which is a marker of the aggressiveness of the cyst itself.


Assuntos
Endometriose/cirurgia , Laparoscopia/efeitos adversos , Cistos Ovarianos/cirurgia , Doenças Ovarianas/cirurgia , Folículo Ovariano/cirurgia , Adulto , Fatores Etários , Biópsia , Distribuição de Qui-Quadrado , Endometriose/patologia , Endometriose/fisiopatologia , Feminino , Fertilidade , Humanos , Itália , Cistos Ovarianos/patologia , Cistos Ovarianos/fisiopatologia , Doenças Ovarianas/patologia , Doenças Ovarianas/fisiopatologia , Folículo Ovariano/patologia , Folículo Ovariano/fisiopatologia , Seleção de Pacientes , Estudos Prospectivos , Medição de Risco , Fatores de Risco , Resultado do Tratamento , Adulto Jovem
20.
Fertil Steril ; 95(5): 1773-6, 2011 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-21300332

RESUMO

OBJECTIVE: To investigate the efficacy of ultrasound-guided transabdominal follicular aspiration when the ovaries are not accessible transvaginally. DESIGN: Retrospective case-control study. SETTING: University-hospital based in vitro fertilization (IVF) clinic. PATIENT(S): 69 women undergoing transabdominal follicular aspiration for oocyte retrieval, including 12 cases of mixed abdominal/vaginal aspiration, compared with controls matched by age, follicle number, and year of procedure undergoing standard transvaginal aspiration. INTERVENTION(S): Transabdominal follicular aspiration when one or more ovaries could not be retrieved via standard transvaginal aspiration. MAIN OUTCOME MEASURE(S): Total and mature oocytes retrieved, damaged oocytes, fertilization rate, embryo number and quality, and clinical and ongoing pregnancy rates. RESULT(S): Cases of transabdominal aspiration had slightly fewer oocytes retrieved, but no statistically significant differences were found for damaged oocytes, fertilization rates, embryo number and quality, or pregnancy rates. In 12 years, one complication requiring hospitalization was noted. CONCLUSION(S): This study demonstrates that transabdominal ultrasound-guided follicular aspiration is safe and efficacious, yielding clinical results in women with significant ovarian displacement, comparable with results achieved by transvaginal aspiration in women with normally positioned ovaries. Transabdominal ultrasound-guided aspiration should be the modality of choice when the ovaries are not accessible transvaginally.


Assuntos
Cavidade Abdominal/cirurgia , Procedimentos Cirúrgicos em Ginecologia/métodos , Recuperação de Oócitos/métodos , Folículo Ovariano/cirurgia , Vagina/diagnóstico por imagem , Adulto , Estudos de Casos e Controles , Feminino , Fertilização in vitro , Humanos , Laparoscopia/métodos , Gravidez , Taxa de Gravidez , Estudos Retrospectivos , Falha de Tratamento , Ultrassonografia
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