RESUMEN
Post-translational modifications of proteins are known to be important in protein activity and ERalpha is known to be phosphorylated at multiple sites within the protein. The exact function of site-specific phosphorylation in ERalpha is unknown, although several hypotheses have been developed using site-directed mutagenesis and cell culture models. Targeting the ERalpha at the level of such post-translational modification pathways would be a new and exciting approach to endocrine therapy in breast cancer, but adequate knowledge is lacking with regard to the relevance of site-specific phosphorylation in ERalpha in human breast cancer in vivo. Recently, antibodies to P-Serine(118)-ERalpha and P-Serine(167)-ERalpha, two major sites of phosphorylation in ERalpha, have become available and some in vivo data are now available to complement studies in cells in culture. However, the in vivo data are somewhat contradictory and limited by the small cohorts used and the lack of standard well-characterized reagents and protocols.
Asunto(s)
Neoplasias de la Mama/metabolismo , Receptor alfa de Estrógeno/metabolismo , Regulación Neoplásica de la Expresión Génica , Serina/metabolismo , Neoplasias de la Mama/patología , Humanos , Fosforilación , Serina/química , Serina/genéticaRESUMEN
The steroid receptor RNA activator (SRA) has previously been characterized as belonging to the growing family of functional non-coding RNAs. However, we recently reported the Western blot detection of a putative endogenous SRA protein (SRAP) in breast cancer cells. Herein, we successfully suppressed the expression of this protein through specific RNA interference assay, unequivocally confirming its existence. Moreover, using database searches and Western blot analysis, we also showed that SRAP is highly conserved among chordata. Overall, our results suggest that SRA is the first example of a new class of functional RNAs also able to encode a protein.
Asunto(s)
Biosíntesis de Proteínas/genética , ARN no Traducido/genética , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Secuencia de Aminoácidos , Animales , Western Blotting , Secuencia Conservada , Células HeLa , Humanos , Datos de Secuencia Molecular , Músculo Esquelético/metabolismo , Interferencia de ARN/fisiología , ARN Largo no Codificante , Ratas , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Alineación de Secuencia , Especificidad de la Especie , TransfecciónRESUMEN
OBJECTIVE: To investigate the current incidence of vasectomy reversal procedures, the techniques used and which practitioners use them. PATIENTS AND METHODS: Using a questionnaire, 130 general surgeons and urologists practising in Merseyside and North Wales were surveyed. RESULTS: The response rate was 74%, with 24 urological surgeons and 14 general surgeons undertaking vasectomy reversal. Annually, urological surgeons carried out significantly more procedures than did general surgeons, at 8.5 and 5.3 (P = 0.029), respectively. They were also more likely to use double-layer closure and microsurgical techniques, whilst significantly less likely to use stents. Urologists reported significantly greater patency rates, at 76% and 52% (P = 0.017), respectively, with no significant differences in subsequent pregnancy rates (30% vs 25%). Only one practitioner checked tubal patency in the female partner before vasectomy reversal. CONCLUSIONS: The use of vasectomy reversal is a cost-effective treatment for men wanting paternity after vasectomy. The technique used by the clinician and proper audit of the results require close attention; it would also appear to be obvious that all the partners of men seeking a vasectomy reversal should have their fertility status established before reversal, something that is clearly not done at present.
Asunto(s)
Vasovasostomía/estadística & datos numéricos , Actitud del Personal de Salud , Criopreservación/estadística & datos numéricos , Inglaterra , Encuestas de Atención de la Salud , Encuestas Epidemiológicas , Humanos , Masculino , Auditoría Médica , Oligospermia , Pautas de la Práctica en Medicina , Stents/economía , Stents/estadística & datos numéricos , Encuestas y Cuestionarios , Vasovasostomía/economía , Vasovasostomía/métodos , GalesRESUMEN
OBJECTIVE: To investigate the effect of the interval between previous vasectomy reversal on retrieval rates of epididymal and testicular spermatozoa using percutaneous epididymal sperm aspiration (PESA), or testicular sperm extraction (TESE), and the subsequent reproductive potential of these gametes in intracytoplasmic sperm injection (ICSI) cycles. PATIENTS AND METHODS: Sixty-six consecutive sperm retrievals were considered in patients who were azoospermic after previous vasectomy, of whom 54 had had a previous failed reversal, the remainder deciding against a reversal. PESA and TESE retrieval rates were noted, as were the time since vasectomy and the interval between vasectomy and unsuccessful reversal. The presence of palpable epididymal cysts was noted, with their effect on sperm retrieval rates. Fertilization and pregnancy rates were analysed in subsequent ICSI cycles using freshly retrieved spermatozoa or frozen-thawed cryopreserved spermatozoa. RESULTS: All 66 patients had sperm retrieved successfully; the success rates for PESA were not significantly affected by previous failed reversal when compared with patients who had not had a reversal, at 14 of 54 (26%) vs five of 12 (P=0.3). The interval since vasectomy did not affect PESA retrieval rates but there was a significantly poorer retrieval rate for PESA in the presence of palpable epididymal cysts, at seven of 35 (20%) vs 12 of 23 (52%) (P=0.012). Fertilization rates were significantly lower using cryopreserved spermatozoa retrieved from either the epididymis or testis (50% vs 70%, P=0.007), although subsequent implantation and pregnancy rates were not significantly different. CONCLUSION: Surgical sperm retrieval is successful in all cases of azoospermia secondary to vasectomy, either by PESA or TESE. There are no clinical markers to indicate which patients will have successful PESA after vasectomy, although the presence of epididymal cysts is associated with significantly lower retrieval rates. The reduction in fertilising ability of cryopreserved spermatozoa does not affect clinical pregnancy rates in ICSI cycles.
Asunto(s)
Espermatozoides , Recolección de Tejidos y Órganos/métodos , Vasovasostomía/métodos , Adulto , Criopreservación/métodos , Fertilización In Vitro/métodos , Humanos , Masculino , Estudios Retrospectivos , Preservación de Semen/métodos , Factores de TiempoRESUMEN
The hypothesis that altered expression of specific coactivators/repressors of the estrogen receptor occurs during human breast tumorigenesis in vivo is examined in this study. Using in situ hybridization and reverse transcription-PCR assays, the expression of two coactivators (SRA and AIB1) and one repressor (REA) of the estrogen receptor was compared between matched breast tumors and adjacent normal human breast tissue. The levels of SRA and AIB1 mRNA were increased in tumors compared with normal tissues (n = 19; Wilcoxon matched pairs test; P < 0.01). In contrast, the expression of REA mRNA was not different between tumors and normal tissues (n = 19; Wilcoxon; P = 0.110). The ratios of AIB1:REA and SRA:REA were higher (Wilcoxon; P < 0.05) in tumors compared with normal tissues. Furthermore, SRA:AIB1 was higher (Wilcoxon; P = 0.0058) in tumors compared with normal tissues. Although our study is small, these data are consistent with the above hypothesis and suggest that such alterations may have a role in the altered estrogen action occurring during breast tumorigenesis.
Asunto(s)
Neoplasias de la Mama/metabolismo , Proteínas de Neoplasias/biosíntesis , ARN no Traducido/biosíntesis , Proteínas Represoras/biosíntesis , Factores de Transcripción/biosíntesis , Mama/metabolismo , Neoplasias de la Mama/genética , Receptor alfa de Estrógeno , Femenino , Humanos , Hibridación in Situ , Proteínas de Neoplasias/genética , Coactivador 3 de Receptor Nuclear , Adhesión en Parafina , Prohibitinas , ARN/biosíntesis , ARN/genética , ARN Largo no Codificante , ARN Mensajero/biosíntesis , ARN Mensajero/genética , ARN Neoplásico/biosíntesis , ARN Neoplásico/genética , ARN no Traducido/genética , Receptores de Estrógenos/fisiología , Proteínas Represoras/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factores de Transcripción/genéticaRESUMEN
Previous studies have shown that lumican is expressed and increased in the stroma of breast tumours. Lumican expression has now been examined relative to other members of the small leucine-rich proteoglycan gene family in normal and neoplastic breast tissues, to begin to determine its role in breast tumour progression. Western blot study showed that lumican protein is highly abundant relative to decorin, while biglycan and fibromodulin are only detected occasionally in breast tissues (n=15 cases). Further analysis of lumican and decorin expression performed in matched normal and tumour tissues by in situ hybridization showed that both mRNAs were expressed by similar fibroblast-like cells adjacent to epithelium. However, lumican mRNA expression was significantly increased in tumours (n=34, p<0.0001), while decorin mRNA was decreased (p=0.0002) in neoplastic relative to adjacent normal stroma. This was accompanied by a significant increase in lumican protein (n=12, p=0.0122), but not decorin. Further evidence of altered lumican expression in breast cancer was manifested by discordance between lumican mRNA and protein localization in some regions of tumours but not in adjacent morphologically normal tissues. It is concluded that lumican is the most abundant of these proteoglycans in breast tumours and that lumican and decorin are inversely regulated in association with breast tumourigenesis.
Asunto(s)
Neoplasias de la Mama/metabolismo , Proteoglicanos/metabolismo , Adulto , Anciano , Biomarcadores de Tumor , Western Blotting , Estudios de Casos y Controles , Progresión de la Enfermedad , Electroforesis en Gel de Poliacrilamida , Femenino , Expresión Génica , Humanos , Hibridación in Situ , Persona de Mediana Edad , Proteoglicanos/genética , ARN Mensajero/análisisRESUMEN
The expression of a specific repressor of estrogen receptor activity (REA) was investigated by a semiquantitative reverse transcription-PCR assay in 40 human breast tumor biopsy samples with respect to steroid hormone receptor status and other known prognostic variables. The data showed that REA expression was positively correlated with estrogen receptor (ER) levels as defined by ligand-binding assays (Spearman r = 03231; P = 0.042) and that the median level of REA mRNA was significantly (Mann-Whitney two-tailed test, P = 0.0424) higher in ER+ tumors (median = 94.5; n = 30) compared with ER- tumors (median = 645; n = 10), with no significant differences (P = 0.4988) associated with progesterone receptor status alone. In addition, REA expression was inversely correlated with tumor grade (Spearman r = -0.4375; P = 0.0054). When the tumors were divided into two groups based on grade, REA expression was significantly (Mann-Whitney two-tailed test, P = 0.0024) higher in low-grade (median = 97; n = 16) compared with high-grade (median = 76; n = 23) tumors. These results provide preliminary data suggesting that the expression of REA varies among breast tumors and is correlated with known treatment response markers and inversely correlated with a marker of breast cancer progression. REA together with ER status may be an improved marker of endocrine therapy responsiveness in human breast cancer.
Asunto(s)
Biomarcadores de Tumor/genética , Neoplasias de la Mama/genética , Carcinoma Ductal de Mama/genética , Receptores de Estrógenos/metabolismo , Proteínas Represoras/biosíntesis , Biopsia , Northern Blotting , Células Cultivadas , Humanos , Prohibitinas , ARN Mensajero/genética , Receptores de Estrógenos/genética , Receptores de Progesterona/genética , Receptores de Progesterona/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Células Tumorales CultivadasRESUMEN
The Mammaglobin gene, a breast-specific member of the uteroglobin gene family, has been previously identified as being overexpressed in some breast tumours, but the cellular origin and relationship to tumour progression are unknown. Using a subtractive hybridization approach, mammaglobin mRNA has also been found to be overexpressed in the in situ compared to the invasive element within an individual breast tumour. Further study by in situ hybridization performed in 13 breast tumours, selected to include normal, in situ, and invasive primary tumour elements, and in most cases axillary lymph node metastases, revealed that mammaglobin expression occurs in all elements, is restricted to epithelial cells, and is significantly increased in tumour cells compared with normal cells ( p< 0.04). Analysis of mammaglobin expression within 20 independent primary breast tumours and their corresponding axillary lymph nodes revealed that all 13 lymph nodes positive and none of the seven nodes negative for metastatic breast carcinoma by histology were mammaglobin-positive by reverse transcription-polymerase chain reaction (RT-PCR) ( p=0.0001). These results suggest that mammaglobin could be a marker of axillary lymph node breast metastases.
Asunto(s)
Biomarcadores de Tumor/genética , Neoplasias de la Mama/genética , Proteínas de Neoplasias/genética , ARN Mensajero/análisis , Uteroglobina/genética , Adulto , Anciano , Anciano de 80 o más Años , Axila , Neoplasias de la Mama/patología , Femenino , Marcadores Genéticos , Humanos , Hibridación in Situ , Metástasis Linfática , Mamoglobina A , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
Previous studies have suggested that elevated serum follicle stimulating hormone (FSH) concentrations are associated with a poor ovarian response to hyperstimulation with human menopausal gonadotrophin (HMG) in in vitro fertilisation (IVF) programmes. We have used the day 2 serum FSH concentration to determine the dose of HMG administered in women under 40 years. If the FSH concentration was below 9 IU/l, a constant dose of 150 IU HMG were administered; if above 9 IU/l a constant dose of 300 IU HMG was used. Women over the age of 40 years were given 300 IU HMG regardless of their serum FSH concentration. This retrospective study was undertaken to assess whether this approach was beneficial for the younger women and also whether the FSH concentration was predictive of outcome in older women. The study included all women < 40 years (n = 143) and > 40 years (n = 32) having their first IVF treatment cycle during 1994. In the younger women, there was no difference in the number of cancelled treatment cycles (9.7% vs. 7.5%); the number of follicles present (9.6 vs. 8.2); serum oestradiol concentration (6971 pmol/l vs. 6686 pmol/l); number of eggs collected (7.9 vs. 5.7); number of embryos created (3.7 vs. 3.6); and pregnancy rate (13.5% vs. 15%) between women with normal (n = 103) or elevated (n =40) FSH concentrations. By using the serum FSH concentration to select women in whom a poor response was expected, and administering a higher dose of HMG, a similar ovarian response was produced and the pregnancy rate was similar to those in women with normal FSH concentrations. Women over 40 years with elevated serum FSH concentrations (n = 17) had a significantly (P < 0.05) higher cancellation rate (17.6% vs. 0%) and fewer number of eggs collected (6.9 vs. 2.5) than the group with normal FSH concentrations (n = 15). One woman conceived in each group. These findings confirmed previous studies showing that the serum FSH is predictive of ovarian response. This study confirmed the value of measuring the day 2 serum FSH concentration as a predictor of response; and it provides a scientific approach to determine the dose of HMG administered for IVF stimulation. A satisfactory response to induction of ovulation will be achieved using 150 IU HMG in women with FSH < 9 IU/l but if the FSH is raised i.e. above 9 IU/l, 300 IU is required to achieve a similar response.
RESUMEN
PURPOSE: The purpose of the present study was (i) to assess the value of using a low dose of hMG (75 IU/day) to achieve ovarian stimulation in women who have previously shown an exaggerated response to a standard dose of 150 IU human menopausal gonadotropin/day in a desensitization (group I) or flare-up (group II) protocol and (ii) to determine whether the choice of GnRH-a regimen in a subsequent cycle, namely, a desensitization or flare-up protocol, influenced the effectiveness of the low dose of hMG. RESULTS: In group I, 75% (12/16) and 57% (8/14) of the subsequent desensitization and flare-up protocols, respectively, were cancelled because of inadequate ovarian response. Similarly, the cancellation rates in group II were 10 of 10 and 7 of 11 (64%), respectively. The total cancellation rate (groups I and II together) with the desensitization protocol was higher than that using the flare-up protocol (P < 0.05). CONCLUSION: The simple use of a reduced dose of hMG (75 IU/day) for subsequent in vitro fertilization in women to minimize the risk of the development of ovarian hyperstimulation is of limited benefit since a large proportion then shows an inadequate response. This is particularly pronounced with a subsequent desensitization protocol which does not utilize endogenous gonadotropins to initiate follicular development.
Asunto(s)
Buserelina/uso terapéutico , Fertilización In Vitro , Menotropinas/uso terapéutico , Síndrome de Hiperestimulación Ovárica/prevención & control , Inducción de la Ovulación , Adulto , Buserelina/efectos adversos , Relación Dosis-Respuesta a Droga , Transferencia de Embrión , Estradiol/sangre , Femenino , Gonadotropinas/uso terapéutico , Humanos , Leuprolida/uso terapéutico , Menotropinas/efectos adversos , Ciclo Menstrual , Síndrome de Hiperestimulación Ovárica/epidemiología , Síndrome de Hiperestimulación Ovárica/fisiopatología , Embarazo , Resultado del Embarazo , Radioinmunoensayo , Factores de RiesgoRESUMEN
The World Health Organization (WHO) laboratory manual (1992) states that assessment of sperm motility can be performed at either 37 degrees C or room temperature (20-24 degrees C). The motility of spermatozoa in 44 semen samples (22 fresh samples and 22 frozen-thawed samples) was assessed at both of these temperatures and a significant difference in the motility profiles was noted, specifically an increase at 37 degrees C in the percentage (expressed here as median and ranges) of spermatozoa with excellent progressive motility and an overall increase in the percentage with total progressive motility. With fresh samples the excellent progressive motility increased from 41 (19-53) to 54 (30-66) and the overall motility from 58.5 (39-74) to 65.0 (40-79). With the frozen-thawed samples the excellent motility increased from 14 (1-33) to 25 (6-45) and the overall motility from 30.5 (14-51) to 33.0 (16-52). As the WHO laboratory manual was published 'In response to a growing need for the standardisation of procedures for the examination of human spermatozoa' it is proposed that only one temperature for routine analysis should be used, namely 37 degrees C, which may have more physiological relevance and eliminate effects of fluctuations in ambient laboratory temperature.
Asunto(s)
Motilidad Espermática/fisiología , Humanos , Masculino , TemperaturaRESUMEN
Since the human acrosome reaction is considered a prerequisite for normal fertilization and the spontaneous acrosome reaction rate is low, laboratory tests using calcium ionophores to induce the acrosome reaction have been devised and applied to the investigation of patients. The introduction of any new laboratory test into routine clinical practice is usually accompanied by the determination of intra- and inter-subject variability within the normal population, and the derivation of reference values to distinguish between affected and unaffected populations. The acrosome reaction to ionophore challenge (ARIC) test was evaluated and found to have (i) intra- and inter-assay coefficients of variation of 10.8 and 18.8% respectively, (ii) a high degree of intra-subject variability for three subjects studied over a 10 week period, (iii) a high degree of inter-subject variability when aliquots of 20 ejaculates of donor semen of proven fertility were tested, and (iv) no effect of length of sexual abstinence on ARIC values. The results of this study suggest that the use of fresh semen samples from subjects of proven fertility for quality control purposes in the ARIC test may be inadequate due to the high degree of intra-subject variability, and that this problem may be overcome by utilizing a frozen quality control sample. The results also suggest that an isolated negative ARIC test is not necessarily indicative of functionally incompetent spermatozoa, and highlight the importance of examination of the normal population prior to the clinical application of such a test.
Asunto(s)
Acrosoma/fisiología , Calcimicina/farmacología , Fertilidad , Abstinencia Sexual , Acrosoma/efectos de los fármacos , Humanos , Masculino , Control de Calidad , Valores de Referencia , Reproducibilidad de los ResultadosRESUMEN
OBJECTIVE: To compare the outcome of in vitro fertilisation (IVF) and gamete intrafallopian transfer (GIFT) cycles in women with or without ultrasound features of polycystic ovary syndrome (PCOS). DESIGN: A consecutive series from January to December 1989. SUBJECTS: Twenty-five women with PCOS scheduled for assisted conception. The controls were 139 women with normal ovaries. SETTING: A single centre specialist fertility unit, Manchester, UK. INTERVENTIONS: Pituitary desensitisation was with buserelin. In the PCOS group ovarian stimulation was with 1 ampoule (75 iu FSH) of hMG/day in 12 women (Group I) and two ampoules/day in 13 (Group II). The controls (Group III) were given two ampoules of hMG daily. Human chorionic gonadotrophin (hCG; 10,000 iu) was given when three follicles measured > or = 20 mm diameter. MAIN OUTCOME MEASURES: Serum oestradiol (E2) concentrations, number of follicles, clinical pregnancies, features of the ovarian hyperstimulation syndrome (OHS). RESULTS: Women with PCOS (Groups I or II) had more follicles > or = 14 mm diameter on the day of the hCG injection (P < 0.005), higher serum E2 concentrations on the day after the hCG (P < 0.05) and more oocytes retrieved (P < 0.05) than the controls. The OHS was more prevalent in those with PCOS (32% versus 6.5%; P < 0.05). The clinical pregnancy rate per embryo transfer (27% versus 22%) or gamete transfer (25% versus 39%) and the rate of spontaneous miscarriage (33% versus 12%) were not statistically different. CONCLUSIONS: The pregnancy rate and outcome of pregnancy following IVF or GIFT in women with or without PCOS are similar. Women with PCOS are at a higher risk of developing OHS.
Asunto(s)
Buserelina/uso terapéutico , Gonadotropina Coriónica/uso terapéutico , Fertilización In Vitro , Transferencia Intrafalopiana del Gameto , Síndrome del Ovario Poliquístico , Embarazo , Adulto , Estradiol/sangre , Femenino , Humanos , Infertilidad Femenina/terapia , Síndrome de Hiperestimulación Ovárica/sangre , Síndrome de Hiperestimulación Ovárica/etiología , Síndrome del Ovario Poliquístico/complicaciones , Síndrome del Ovario Poliquístico/fisiopatología , Resultado del Embarazo , Factores de RiesgoRESUMEN
OBJECTIVES: To analyse the incidence and factors associated with the ovarian hyperstimulation syndrome (OHS) in our IVF/GIFT programme before and after the introduction of a strategy to cryopreserve all embryos from women judged to be at risk. DESIGN: Two hundred forty-one consecutive IVF/GIFT cycles from January to December 1989. SETTING: Specialist fertility unit, Manchester, UK. INTERVENTIONS: Pituitary suppression was effected by a daily subcutaneous injection of buserelin (500 micrograms) beginning 7 days before the expected menses. The ovarian stimulation was with variable amounts of human menopausal gonadotrophin. Ovulation was induced with 10,000 i.u. human chorionic gonadotrophin (hCG). From January to May (period A), gametes/embryos were replaced and 2000 i.u. hCG given, irrespective of the serum oestradiol (E2) concentration. From June to December (period B), all the embryos from women with an E2 > 3500 pg/ml on the day of ovulatory trigger were electively cryopreserved. MAIN OUTCOME MEASURES: Serum E2, features of moderate or severe OHS, clinical pregnancies. RESULTS: The OHS occurred in 10/105 (9.5%) and 12/136 (8.8%) cycles in periods A and B, respectively. Fewer women (6% versus 60%, P < 0.05) who had their embryos cryopreserved developed severe OHS compared with women with an E2 > 3500 pg/ml who became pregnant after gamete/embryo transfer in period A. The main factors associated with the development of OHS were serum E2 concentrations > 3500 pg/ml, whether gamete/embryos were replaced and the additional hCG given, the occurrence of a pregnancy and the presence of polycystic ovary disease. CONCLUSION: The elective cryopreservation of all embryos from women with high E2 levels reduced the severity, but not the incidence of symptomatic OHS.
Asunto(s)
Criopreservación , Embrión de Mamíferos , Síndrome de Hiperestimulación Ovárica/prevención & control , Pamoato de Triptorelina/análogos & derivados , Adulto , Buserelina/uso terapéutico , Estradiol/sangre , Femenino , Fertilización In Vitro , Transferencia Intrafalopiana del Gameto , Hormona Liberadora de Gonadotropina/efectos adversos , Hormona Liberadora de Gonadotropina/análogos & derivados , Humanos , Incidencia , Infertilidad Femenina/sangre , Infertilidad Femenina/terapia , Persona de Mediana Edad , Síndrome de Hiperestimulación Ovárica/sangre , Síndrome de Hiperestimulación Ovárica/inducido químicamente , Embarazo , Factores de RiesgoRESUMEN
From 1st June 1989 to 31st May 1991, 78 women with a serum oestradiol level greater than 3500 pg/ml on the day of the ovulatory trigger, following pituitary suppression with buserelin and ovarian stimulation with human menopausal gonadotrophins (HMG), had all their embryos electively cryopreserved at the pronucleate stage to minimize the risk of developing ovarian hyperstimulation syndrome (OHS). Treatment with buserelin was continued in the luteal phase. A median of 19 oocytes (range 7-43) was obtained and 12 embryos (range 1-37) frozen per cycle. Twenty-one (27%) women developed OHS (six severe). Women developing OHS had higher (P less than 0.05) serum oestradiol concentrations on the 7th day after oocyte retrieval, compared to those who did not. No differences were found for any of the following criteria: aetiology of infertility, age, total dose of HMG, number of oocytes, fertilization rate or freeze-thaw survival of embryos. Subsequently, 125 frozen-thawed embryo replacements have been undertaken, using buserelin and hormone replacement therapy (HRT) (n = 93) or natural cycles (n = 32). The overall freeze-thaw survival and implantation rates per embryo were 71.8 and 11.7%, respectively. The pregnancy rates in natural cycles (19%) and buserelin/HRT cycles (29%) were not significantly different.
Asunto(s)
Buserelina/uso terapéutico , Criopreservación , Implantación del Embrión/fisiología , Embrión de Mamíferos , Síndrome de Hiperestimulación Ovárica/terapia , Supervivencia Celular/efectos de los fármacos , Estradiol/uso terapéutico , Femenino , Humanos , Fase Luteínica/efectos de los fármacos , Oocitos/efectos de los fármacos , Síndrome de Hiperestimulación Ovárica/diagnóstico , Embarazo , Resultado del Embarazo , Progesterona/uso terapéutico , Factores de Riesgo , Resultado del TratamientoAsunto(s)
Criopreservación/métodos , Embrión de Mamíferos , Edad Materna , Resultado del Embarazo , Adulto , Femenino , Humanos , EmbarazoRESUMEN
Supernumerary embryos following treatment by IVF or GIFT were cryopreserved at the pronucleate, early cleavage or expanded blastocyst stages. The success of embryo cryopreservation at these stages was evaluated in terms of (i) the proportion of embryos surviving the freeze/thaw procedure; (ii) the proportion of patients reaching embryo replacement; and (iii) the incidence of pregnancy per replacement. Significantly more embryos survived when frozen/thawed at the pronucleate (44/61; 72%) or early cleavage stages (48/80; 60%), than at the expanded blastocyst stage (13/34; 38%). A significantly higher proportion of patients had embryo replacements when embryos were frozen/thawed at the pronucleate (17/19; 89%) or early cleavage stages (21/24; 88%), than at the expanded blastocyst stage (9/17; 53%). Following replacement of frozen/thawed pronucleate and early cleavage stage embryos, clinical pregnancy rates of 8/17 (47%) and 3/21 (14%) clinical pregnancies were achieved, respectively. No pregnancies were achieved following replacement of frozen/thawed expanded blastocysts.
Asunto(s)
Blastocisto , Fase de Segmentación del Huevo , Criopreservación/métodos , Transferencia de Embrión/métodos , Buserelina/análogos & derivados , Buserelina/farmacología , Clomifeno/farmacología , Crioprotectores , Estudios de Evaluación como Asunto , Femenino , Fertilización In Vitro/métodos , Transferencia Intrafalopiana del Gameto/métodos , Goserelina , Humanos , Menotropinas/farmacología , Ovario/efectos de los fármacos , Embarazo , Propilenglicol , Glicoles de PropilenoRESUMEN
Gamete intra-Fallopian transfer (GIFT) was performed in 130 treatment cycles over a 17-month period. In 91% (118/130) of the cycles one or more oocytes were available for insemination in vitro and only GIFT cycles with supernumerary oocytes were included in the present study. Pituitary and ovarian suppression was achieved with buserelin followed by stimulation of multifollicular development by human menopausal gonadotrophin (HMG). Failure of supernumerary oocytes to fertilize was associated with a significantly reduced pregnancy rate (3/23; 13%) compared to cycles where fertilization occurred in vitro (35/95; 37%). These findings demonstrate that the outcome of IVF of supernumerary oocytes may be of particular diagnostic value in couples where the female partner has not conceived following treatment by GIFT after pituitary down-regulation with buserelin and ovarian stimulation with HMG.
Asunto(s)
Fertilización In Vitro , Transferencia Intrafalopiana del Gameto , Infertilidad/terapia , Oocitos/fisiología , Adulto , Buserelina/farmacología , Buserelina/uso terapéutico , Femenino , Humanos , Infertilidad/fisiopatología , Masculino , Menotropinas/farmacología , Menotropinas/uso terapéutico , Ovario/efectos de los fármacos , Ovario/fisiopatología , Hipófisis/efectos de los fármacos , Hipófisis/fisiopatología , EmbarazoRESUMEN
The cryopreservation of semen used in assisted reproduction procedures was carried out exclusively by a simplified method in which a mixture of semen and cryoprotectant was contained in 1-ml tuberculin syringes and plunged directly into liquid nitrogen. Donor semen samples halved and frozen in syringes and in straws in a controlled-rate freezer showed no significant difference in post-thaw motility (P = 0.217) or survival (P = 0.217) after 30 min. However, after 180 min the survival rate showed a significant reduction in syringes (P = 0.045). A significant difference (P less than 0.00008) in the rate of fertilization of oocytes was seen in IVF cycles using frozen-thawed donor sperm (58/142, 42%) when compared to fresh sperm from husbands (2315/3926, 59%). A significant reduction (P less than 0.00005) in fertilization rate was also observed in the case of supernumerary oocytes in GIFT cycles with the cryopreserved donor sperm (29/132, 22%) compared to the husbands' sperm (239/514, 46%). However, the pregnancy rate following IVF and embryo replacement was the same after fertilization with fresh sperm (75/351, 21%) as opposed to frozen sperm (3/14, 21%). Furthermore, a higher pregnancy rate was observed in GIFT with frozen donor sperm (9/19, 47%) than with fresh sperm from husbands (28/103, 27%), though this was not statistically significant (P = 0.079). These results show this simplified methods of semen cryopreservation to be effective when used in an IVF and GIFT programme, giving pregnancy rates comparable to fresh normospermic semen samples. The method is simple, quick and inexpensive.