Translocation t(1;5) in a case of carcinoma of the cervix.

We report a case of carcinoma of the cervix uteri, which presented both numerical and structural chromosome changes. The tumor showed the coexistence of lines with different modal chromosome numbers, but all of them with the t(1;5)(q25;132). We also observed the presence of double minutes, dicentric chromosomes, small acentric fragments, and/or tri- and quadriradial figures in 11% of the cells.

[Pregnancy with spermatozoa from the head of the epididymis in spermatic duct agenesis]. / Embarazo logrado con espermatozoides obtenidos de la cabeza epididimaria en una agenesia de vías espermáticas.

From a male affected of agenesis of corpus and cauda epididymis and vas deferens, sperms were surgically obtained aspirating epididymal content with the help of a surgical microscope. Motile sperms were separated after discontinuous Percoll gradient centrifugation and used to inseminate in vitro the spouse's oocytes 8 hrs. after aspiration according to the current techniques. Twenty hours later, oocytes were checked for fertilization and transferred to fresh culture medium. From 13 preovulatory oocytes only one was fertilized and transferred to the Fallopian tube at the 6 cells stage. Fourteen days after embryo transfer, serum HCG concentration was 320 mU/ml and on day 29th a 5mm fetal sac showing heart beats was detected by transvaginal sonography. After an uneventful 38 weeks gestation a normal baby girl weighing 2,800 g was delivered.

[Pregnancy obtained by intracytoplasmic oocyte injection of spermatozoa retrieved from a cryopreserved testicular biopsy]. / Embarazo logrado por inyección intracitoplasmática de ovocitos con espermatozoides provenientes de una biopsia de testículo criopreservada.

We have obtained a clinical pregnancy in a 26 year old patient by means of in vitro oocyte fertilization by intracytoplasmic injection of spermatozoa obtained from a cryopreserved testicular biopsy. In a first attempt performed with fresh biopsy material, the woman became pregnant, but the pregnancy ended in a spontaneous abortion. In a second cycle, the spermatozoa were retrieved from a cryopreserved sample saved from the first attempt. Twelve metaphase II oocytes were collected and injected; from these, nine became fertilized, three preembryos were transferred at the eighth-cell stage and the other six were cryopreserved. An ongoing clinical pregnancy was obtained with two gestational sacs.

[Treatment with enoxaparin adapted to the fertility programs in women with recurrent abortion and thrombophilia]. / Tratamiento con enoxaparina adaptado a los programas de fertilidad en mujeres con aborto recurrente y trombofilia.

Acquired and inherited thrombophilia are associated with recurrent pregnancy loss (RPL). Antithrombotic therapy could restore hemostatic balance and improve early placentation and gestational outcome. We evaluated the efficacy of enoxaparin adapted to the fertility program for prevention of pregnancy loss in 35 women (W) with early RPL and thrombophilia. Previous to the diagnosis of thrombophilia, they had had a total of 105 gestations of which 89 (85%) ended in early pregnancy loss. After diagnosis of thrombophilia, 35 subsequent pregnancies were treated with enoxaparin. In 5 cases assisted reproductive techniques were necessary to achieve pregnancy due to couple infertility. In 17 W who had had at least one preclinical pregnancy loss, enoxaparin (20 mg/d/s.c.) was started previous to conception and adapted to the fertility program. All the women continued with the gestational regime. Eighteen W with only clinical pregnancy loss started enoxaparin (20 mg twice per day s.c.) after biochemical pregnancy diagnosis. During gestations heparin dose was adjusted with anti Xa test, maintaining a range between 0.3 at 0.6 u/ml. With antithrombotic therapy, 30/35 (85%) of the pregnancies ended in live birth versus 16/105 (15%) of the pregnancies without treatment (p < 0.001). These results suggest that enoxaparin adapted to the fertility program can be effective in the prevention of preclinical and clinical abortion in women with thrombophilia.

Clinical practice of embryo transfer.

In this review, several embryo transfer methods are considered, together with factors involved in achieving an effective transfer. The approach most used is transcervical intrauterine transfer. This is described in detail, together with the many variables influencing success, e.g. technical ability and training of personnel, catheter choice, value of a previous 'dummy transfer' and the need to minimize trauma during transfer and so prevent damage to the uterine lining, bleeding and uterine contractions. These factors can each negatively impact on pregnancy rates. Emphasis is put on quality, developmental stage and number of embryos to be transferred to limit multiple pregnancies and their unwanted side-effects. Culture to blastocyst stages and single embryo transfer when optimal quality embryos are available are discussed as means of avoiding multiple pregnancies. Reference is made to embryo cryopreservation and fertility following frozen embryo transfer. Other techniques, such as ultrasound-controlled transcervical intrauterine transfer, and ultrasound-controlled transmyometrial transfer, are reviewed. More invasive procedures, generically grouped as surgical embryo transfer, including gamete intra-Fallopian transfer (GIFT), zygote intra-Fallopian transfer (ZIFT), pronuclear stage transfer and embryo intra-Fallopian transfer (EIFT), are also described. These techniques had a place in IVF when the need to apply assisted reproductive techniques exceeded the capacity of most laboratories, but not today thanks to refined laboratory technology and improved understanding of implantation. Alternative assisted reproductive technologies, such as direct intra-follicular insemination (DIFI), Fallopian spermatic perfusion (FSP), peritoneal oocyte stage and sperm transfer and intra-vaginal culture (IVC), are mentioned briefly.

Birth resulting from transfer of blastocysts cryopreserved with propanediol after spontaneous hatching.

This paper reports the birth of a healthy baby resulting from transfer of blastocysts that were cryopreserved using propanediol after spontaneous hatching. A young infertile couple underwent IVF treatment in the clinic. After several IVF attempts, two births resulted; the first one with fresh embryos in 1996 after three IVF cycles, and the second one in 1999 (after a new IVF cycle in 1998) with frozen blastocysts that had remained cryopreserved in 1.5 mol/l propanediol and 0.1 mol/l sucrose after spontaneous hatching. This report of a healthy baby following transfer of hatched blastocysts frozen in propanediol supports further exploration of this approach.

Empty follicle syndrome due to human errors: its occurrence in an in-vitro fertilization programme.

We report five cases in which no oocytes were retrieved after standard ovarian stimulation for in-vitro fertilization (IVF), and in which it was found that mistakes had been made at the time of human chorionic gonadotrophin (HCG) administration. In all five cases, oocyte retrieval was achieved after injecting HCG, when necessary, and reprogramming aspiration 24-36 h later. A mean of 7+/-3.2 MII oocytes were recovered per patient and 3.2+/-0.8 embryos were transferred. Three clinical pregnancies were obtained, and four healthy infants were born. In our programme, these were the only cases of empty follicle syndrome (EFS) that appeared over a total of 1118 cycles, and were all explained by human error in the administration of HCG. Our experience shows that human error could be considered a significant factor in the aetiology of empty follicle syndrome, and that EFS may be in part avoided by taking simple preventive measures.

[Fertile amenorrhea associated to diabetes insipidus]. / Amenorrea fértil asociada a diabetes insípida

A case of diabetes insipidus is presented which appeared in a 13 year old girl associated with hormone-resistant amenorrhea; she went through two normal pregnancies and partutition at 22 and 25, indicating a fertile amenorrhea. During a total of 17 years of observation the amenorrhea persisted, with the exception of a few normal menstruation periods at the beginning of the disease. She remained permanently under treatment with pitressin tannate. Repeated administrations of estrogens, gestagens and chorionic gonadotrophin, had no effect. An endometrial biopsy revealed a presecretory phase. Acidophilic index in vaginal smears as well as serial determinations of urinary pregnanodiol indicated cyclic changes. Daily determinations of urinary pregnanodiol indicated cyclic changes. Daily determinations of plasma gonadotrophins during 28 days revealed normal levels, with normal FSH pulse and ovulatory type peak of LH. An LH-RH test gave marked and characteristic increase of both hormones. The data indicate the integrity of the hypothalamo-hypophyso-ovaric system, with cyclic changes and formation of corpus luteum, vaginal trophism and endometrial changes, concordant with the two normal pregnancies. In this case, the amenorrhea can only be explained by alteration of the usual endometrial vascular changes. The coexistence of diabetes insipidus and fertile amenorrhea is discused in relation with the possible participation of vasopressin in the mechanism of menstruation.

Capacity of adult and prepubertal mouse oocytes to undergo embryo development in the presence of cysteamine.

The present study was carried out to study de novo glutathione (GSH) synthesis and to evaluate the effect of stimulating GSH synthesis during in vitro maturation (IVM) of adult and prepubertal mouse oocytes on the embryo developmental rate. Adult (8 weeks old) and prepubertal mice (24-26 days old) were primed with 5 IU of PMSG and oocytes were retrieved from the ovary 48 hr later for IVM. After IVM (18 hr) Cumulus oocyte complexes (COC) were in vitro fertilized (IVF) and in vitro culture (IVC) in order to observe embryo development. The IVM medium was supplemented with: 0, 25, 50, 100, or 200 microM of cysteamine. To study the novo GSH synthesis, 5 mM BSO was added during IVM of adult or prepubertal oocyte. Developmental rates up to blastocyst were recorded for each group. Experiments also included a group of ovulated oocytes (in vivo matured) after priming with PMSG and HCG. After IVM of adult mice oocytes, an improvement was observed on embryo development in all the supplemented groups when compared with the untreated group (P < 0.05). No differences were observed in blastocyst rate among IVM oocytes with cysteamine and ovulated oocytes. Prepubertal IVM mouse oocytes had a lower cleavage rate compared with ovulated oocytes (P < 0.05). Cysteamine failed to improve prepubertal oocytes developmental rates (P > 0,05). 2-cell embryos, coming from IVM prepubertal oocytes and ovulated oocytes had the same preimplantation developmental rate up to the blastocyst stage. In prepubertal, and adult oocytes an inhibition of embryo development was observed when buthionine sulfoximide (BSO), a specific inhibitor of the gamma-glutamylcysteine synthetase, was added during oocyte maturation (P < 0.01). In conclusion, an improvement in mouse embryo development was observed when cysteamine was added to the IVM medium of adult mice oocytes. In prepubertal oocytes cysteamine addition during oocyte maturation failed to improve embryo developmental rates. The presence of BSO lowered or completely blocked blastocyst development. This proves that, de novo GSH synthesis during oocyte maturation of adult and prepubertal oocytes undoubtedly plays an important role in embryo development. The improvement on oocyte competence observed in adult mice oocytes is probably related to intracellular GSH synthesis stimulated by cysteamine. Nevertheless the reason why cysteamine failed to improve prepubertal oocytes competence remains as an open question.

Cysteamine supplementation during in vitro maturation and embryo culture: a useful tool for increasing the efficiency of bovine in vitro embryo production.

Cysteamine when added during in vitro maturation (IVM) or in vitro embryo culture (IVC) stimulates glutathione (GSH) synthesis and improves embryo developmental rates. This suggests that GSH synthesis is decreased in the in vitro produced embryo. The present study was carried out to evaluate if addition of cysteamine to culture medium at the same time, during IVM and IVC of bovine oocytes, may promote an overall improvement on the developmental rate and embryo quality. Oocytes were matured in TCM 199 supplemented with 10% (v/v) fetal calf serum, hormones, and 0 or 100 microM of cysteamine for 24 hr. After IVM, the oocytes were fertilized (day 0). Day 2 embryos (2-8 cell) were washed and transferred to fresh IVC medium supplemented with 0, 25, 50, or 100 microM of cysteamine and cultured for 48 hr. After this, embryos were cultured in IVC medium without cysteamine until day 8 of IVC. In the present study, we confirmed our previous results by demonstrating that the percentage of embryos that developed to the blastocyst stage was significantly higher (P < 0.05) when 100 microM of cysteamine was added during IVM, and this was further improved when 100 and 50 microM of cysteamine where present during IVM and IVC, respectively (P < 0.05). After cryopreservation, no differences were observed on embryo development, but a significant increase on embryo hatching was found between unsupplemented and supplemented oocytes with 100 and 50 microM of cysteamine during IVM and IVC, respectively (P < 0.05). We can conclude that GSH synthesis stimulation during bovine IVM with cysteamine, concomitant with GSH stimulation during IVC, will be a useful and simple tool for increasing the efficiency of in vitro bovine embryo production.

Circulating immune complexes in breast cancer patients

El objetivo de este trabajo fue evaluar la presencia de complejos inmunes circulantes (CIC) en el suero de pacientes con cáncer de mama, Los CIC de 146 pacientes con cáncer de mama se detectaron por el método de combinación a I-Clq. Se consideraron como valores elevados aquellos que excedieron la media aritmética (x+2DS) de los valores obtenidos en el grupo control (11%). Los valores elevados de combinación a Clq radioactivo fueron detectados antes del tratamiento en los 3 grupos estudiados con la siguiente incidencia: N0: 42%; N1: 67%; N2: 86%. En los primeros 12 meses post-tratamiento los niveles de CIC descendieron en los 3 grupos: N0: 18%; N1: 9%; N2: 23%. Sin embargo, a partir de los 12 meses post-tratamiento la incidencia fue en N0: 14%; N1: 62%; N2: 78%. Estos resultados fueron comparados con los valores obtenidos en el suero de 45 pacientes con displasia de mama

Embarazo con espermatozoides testiculares / Pregnacy with testicular spermatozoa

En un paciente con diagnóstico de agenesia de vías espermáticas se propuso afectuar una aspiración epididimaria asociada a FIV (fertilización in vitro). En la exploración excrotal no se encontró epidídimo (debido probablemente a antecedente quirúrgico de biopsia epididimaria) por lo que se efectuó una biopsia testicular. Este material fue procesado obteniéndose escasos espermatozoides (algunos cientos). Estos espermatozoides fueron inyectados en el citoplasma de los ovocitos de la esposa (técnica de ICSI).De los 18 ovocitos inyectados, 3 se dañaron durante el procedimiento y a las 16 horas se observaron signos de fertilización normal en 6 de ellos, prosiguiendo el desarrollo 4 de embriones que fueron transferidos al útero por vía transcervical, 3 en estado de 8 células y uno de 6 células, habiéndosele efectuado previamnete en la zona pelúcida de cada uno de los embriones un orificio de 20 micrones con solución ácida de Tyrode. A los 15 días de la transferencia (2-1-95) el resultado de la subunidad beta de HCG fue positiva, detectándose al mes (2-2-95) por ecografía un embrión de 19 mm, cursando un embarazo normal

Embarazo logrado con espermatozoides obtenidos de la cabeza epididimaria en una agenesia de vías espermáticas / Pregnancy with spermatozoa from the head of the epididymis in a spermatic ducts agenesis

De un hombre afectado de agenesia de cuerpo y cola de epidídimos y conductos deferentes, se obtuvieron espermatozoides mediante la aspiración microquirúrgica del contenido epididimario. Los espermatozoides móviles se separaron de las muestras aspiradas mediante centrifugación a través de un gradiente discontínuo de Percoll, y se procedió a la fertilización in vitro de los oocitos obtendios de la esposa con la técnica habitual. La inseminación se realizó a las 8 horas de la recolección de los oocitos. Veinte hora después se examinaron los oocitos para detectar signos de fertilización transferiéndolos a medio de cultivo fresco. De 23 oocitos preovulatorios, sólo fertilizó uno, el cual fue transferido a la trompa de Falopio al alcanzar el estadío de 6 células. A los 14 días de la transferencia se detectó en sangre una subunidad HCG de 320 mU/ml y el día 29 se visualizó por ecografía transvaginal un saco fetal de 5 mm con actividad cardíaca. Después de una gestación de 38 semanas se produjó el parto de una niña normal de 2 800 g de peso

Efecto de la estimulación de la síntesis de glutation durante la maduración de oocitos de bovinos y cultivo de los embriones in vitro en la tasa de desarrollo y calidad de los embriones / Effect of stimulation of glutathione synthesis during in vitro maturation of bovine oocytes and in vitro embryo culture on the development rate and embryo quality

Objetivo: El desarrollo de un sistema de maduración (MIV) y cultivo (CIV) in vitro de oocitos humanos es importante. El uso de oocitos humanos en investigación es problemático. Los oocitos de bovinos han sido propuestos como el modelo más conveniente. Se llevaron a cabo experimentos donde se evaluó el efecto que tiene la estimulación de la síntesis de glutation (GSH) durante la MIV y el CIV sobre el desarrollo embrionario y la calidad de los mismos. Materiales y Métodos: Los oocitos provenientes de ovarios de matadero fueron madurados, fertilizados, cultivados y congelados in vitro. La síntesis de glutation fue estimulada con cisteamina. La tasa de desarrollo y calidad de los embriones se estudió en 5 grupos: MIV y CIV (Día 2, embriones de 2 a 6 células) sin suplementación de cisteamina (Cist) (Grupo Control) (A); MIV suplementado con 100 mM de Cist (MIV-100) y el CIV sin suplementación (B); MIV-100 y CIV suplementado con 25 µM de Cist (C); o 50 µM de Cist (D); o con 100 µM de Cist (E). Se realizaron 7 réplicas con 1.374 oocitos. Los datos transformados se analizaron mediante ANOVA y test de Tukey. Resultados: El desarrollo de los embriones en los grupos B, C y D fueron significativamente superiores al grupo control (A). El grupo D fue el mejor (P<0.05). Además, el grupo D presentó los mejores resultados de sobrevida y eclosión embrionaria luego del congelamiento, comparado con el grupo A. Discusión: Los resultados demuestran que la estimulación de la síntesis de GSH durante la MIV y el CIV de oocitos bovinos mejora las tasas de desarrollo de los embriones y su calidad. Estos resultados nos muestran el papel preponderante que cumple el metabolismo del GSH durante la maduración citoplasmática y el desarrollo de los embriones