Valoración nutricional de escolares de tres países iberoamericanos: Análisis comparativo de las referencias propuestas por el International Obesity Task Force (IOTF) y la Organización Mundial de la Salud Nutritional valuation of schoolchildren from three Ibero-American countries: / Nutritional valuation of schoolchildren from three Ibero-American countries: A comparative analysis of the references proposed by International Obesity Task Force (IOTF) and World Health Organization (WHO)

Introducción: El índice de masa corporal (IMC) resulta un indicador útil para valorar la condición nutricional en la infancia y adolescencia. Si bien no existe consenso respecto a los valores o puntos de corte de IMC que deben utilizarse para la diagnosis del sobrepeso, la obesidad o la insuficiencia ponderal, las referencias propuestas por la Organización Mundial de la Salud (OMS) y la International Obesity Task Force (IOTF) son las más aceptadas. No obstante, existen discrepancias importantes en los resultados obtenidos en función de una u otra referencia. Objetivo: Analizar y comparar la condición nutricional de una muestra internacional a partir del IMC, mediante la aplicación simultánea de las referencias recomendadas por la IOTF y la OMS. Métodos: Se calculó el IMC de 1289 escolares de 10 a 13 años de México, Venezuela y España. A cada niño le fue atribuida una categoría nutricional (bajo peso, normopeso, sobrepeso y obesidad) utilizando el criterio de la IOTF (puntos de corte equivalentes a 18,5; 25 y 30 en adultos) y OMS (-1 DE: bajo peso + 1 DE: sobrepeso y +2 DE: obesidad). Las prevalencias de estado nutricional obtenidas por cada criterio fueron comparadas mediante la prueba de Chi-Cuadrado. Resultados: El porcentaje de escolares en cada una de las categorías nutricionales varió significativamente en función de la referencia empleada. La prevalencia de malnutrición por exceso o por defecto fue, en ambos sexos y en los tres países, superior al emplear los parámetros OMS. La referencia OMS evidenció un 10,9% menos de niños normonutridos y un 13,1% más de malnutridos que la IOTF. Discusión: Los resultados de esta investigación permiten inferir que la referencia propuesta por la OMS resulta ser más "preventiva" al momento de identificar individuos malnutridos. Se subraya la importancia de especificar los criterios empleados para la determinación del estado nutricional en los estudios de carácter epidemiológico, así como de contrastar las referencias empleadas para el diagnóstico. Conclusiones: Las referencias OMS para IMC sobrestiman la prevalencia de bajo peso, sobrepeso y obesidad en relación a los puntos de corte propuestos por el IOTF para el diagnóstico de la malnutrición (AU)

Introduction: Body Mass Index (BMI) is a useful tool for assessing nutritional status in childhood and adolescence. Although there is no consensus on BMI values or cutoffs to be used for the diagnosis of overweight, obesity and underweight references proposed by World Health Organization (WHO) and International Obesity Task Force (IOTF) are the most accepted. However, there are significant discrepancies in the results obtained in terms of either reference. Objective: To analyze and compare the nutritional status of an international sample from IMC, by simultaneous application of IOTF and WHO recommended references. Methods: 1289 schoolchildren’s BMI between 10 and 13 years of Mexico, Venezuela and Spain was estimated. Each child was assigned a nutritional category (underweight, normal weight, overweight and obese) using the IOTF criteria (points cut equivalent to 18,5, 25 and 30 in adults) and WHO (-1 SD: low weight +1 DE: overweight and +2 SD: obesity). The prevalence of nutritional status of each criterion was compared using the Chi-square test. Results: The percentage of students in each of the nutritional categories mottled significantly depending on the reference used. The prevalence of excess or defect malnutrition, in both sexes and in all three countries, was exceeded using the WHO parameters. WHO reference showed 10,9% less than well-nourished children and 13,1 % more malnourished than the IOTF. Discussion: The results of this research allow inferring that the reference proposed by WHO appears to be more "protecting" when identifying malnourished individuals. The importance of specifying the criteria for determination of nutritional status in epidemiologic studies, as well as to contrast the references used for diagnosis, is emphasized. Conclusions: The WHO BMI references overestimate the prevalence of underweight, overweight and obesity in relation to the cutoff points proposed by the IOTF for diagnosing malnutrition (AU)

Electrolyzed water as novel technology to improve hygiene of drinking water for dairy ewes.

Tap water alone (TW) or treated with 3% of slightly acidic electrolyzed water (SAEW) were used in this experiment to study its effect on water quality, blood biochemical parameters and milk yield and composition. Each type of water was supplied to one group of 10 milking ewes for 25 days. Weekly water samples from troughs were taken. On days 1, 12 and 25, milk yield was measured, and milk and blood samples were taken. SAEW reduced (P < 0.05) bacterial counts (aerobic mesophilic, total coliform and streptococcus). Blood gases, biochemical parameters and milk yield and its composition were not affected (P > 0.05). SAEW can be used at 3% rate as a powerful and economic agent for sanitizing drinking water for dairy ewes with no effects on animal performance.

Focal spermatogenesis originates in euploid germ cells in classical Klinefelter patients.

BACKGROUND: Klinefelter syndrome is the most frequent chromosome abnormality in human males. This paper aims to investigate the ploidy of meiotic and pre-meiotic germ cells found in spermatogenic foci, and furthermore, the sex chromosome constitution of Sertoli cells which surround these germ cells in non-mosaic Klinefelter patients. METHODS AND RESULTS: A survey of 11 adult patients diagnosed with classical, non-mosaic Klinefelter syndrome who underwent testicular biopsies, showed that six of them had spermatogenesis foci. The topographical study of the biopsies showed that tubuli with germ cells are a minor fraction (8-24%) of all tubuli, although the overwhelming majority is devoid of germ cells. Using fluorescence in situ hybridization (FISH) with probes for the X-centromere and immunolocalization of meiotic proteins, the present work shows that all the 92 meiotic spermatocytes analyzed with FISH were euploid, 46,XY, and thus can form normal, haploid gametes. On the other hand, Sertoli cells show two marks for the X chromosome, meaning that they are 47,XXY. CONCLUSIONS: These results provide a rationale for the high rate of success in the testicular sperm extraction plus ICSI procedures when applied to Klinefelter patients. It is also in agreement with previous studies in the XXY-mouse model. These spermatogenic foci most probably originate from clones of spermatogonia that have randomly lost one of the X chromosomes, probably during periods of life when high spermatogonial mitotic activity occurs.

Comparación del dolor secundario a lipoaspiración tradicional versus lipolisis láser. Estudio prospectivo / Postoperative pain in suction assited lipoplasty versus laser lipolisis. Comparative prospective study

La lipoaspiración, técnica tradicional usada para el tratamiento de las lipodistrofias con cicatrices mínimas, ha sido a través de los años mejorada con los avances anestésicos, farmacológicos y de instrumental. A su vez han aparecido otras técnicas que han publicitado ventajas respecto al tiempo de recuperación, sintomatología y resultados estéticos. Una de estas técnicas es la laserlipolisis a la que se atribuye menos dolor, menos equimosis y rápido postoperatorio. No se encuentran trabajos que acrediten estas ventajas, por lo que decidimos estudiar si existen ventajas de laserlipolisis en comparación con la lipoaspiración tradicional respecto del dolor, síntoma importante a considerar en el postoperatorio. Para esto se diseñó un trabajo prospectivo randomizado con una muestra de 60 pacientes a los cuales se aplicaron las dos técnicas en diferentes áreas y al mismo paciente. Se evaluó el dolor con escala numérica analógica a las 4 horas, 48 horas y 5 días de postoperatorio. Los resultados mostraron que la laserlipolisis presenta menos dolor en los muslos medido a las 48 horas y 5 días de la intervención. No hay diferencias entre estas dos técnicas en las otras zonas medidas como abdomen, áreas pretrocantéreas (“alforjas”), zona lumbar y pectoral. Considerando todos los pacientes sin diferenciar la técnica usada, las “alforjas” a las 4 horas duelen menos que el abdomen y a los 5 días duelen menos los muslos que el abdomen. No encontramos relación entre volumen aspirado e intensidad del dolor (AU)

The traditional suction-assisted lipoplasty, is a well established and commonly performed technique used for the treatment of lipodystrophias, which has undergone improvements with advances in anesthetics, pharmacology and introduction of new instruments. New techniques have appeared, promising advantages in recovery periods, symptomathology and aesthetic results when compared to the traditional suction- assisted lipoplasty. One of these newly introduced techniques is the laser-assisted lipoplasty (laserlipolysis), which claims less pain, less bruising and faster postoperative recovery. No evidence has been published to support these advantages when comparing laser-assisted lipoplasty to the traditional method, specifically when refering to pain, an important symptom to be taken into consideration in the postoperative period. For this reason we designed a prospective, randomized clinical study comprising 60 patients to which the two techniques were applied to different body areas. Pain was assessed using the numeric analog scale at 4 and 48 hours, and at 5 days after surgery. Evidence showed that laserlipolysis results in less pain of the thighs measured at 48 hours and 5 days after surgery. No difference in pain was found between both techniques on the other body areas evaluated (abdomen, trochanteric bulge, lumbar and pectoral areas). Assesing all patients, independent of the technique used, trochanteric bulge at 4 hours was found to be less painful than the abdominal area, and at 5 day evaluation the thighs were less painful than the abdominal area. No relationship was found between pain intensity and lipoaspirated volume

Decreased protein tyrosine phosphorylation and membrane fluidity in spermatozoa from infertile men with varicocele.

Varicocele is a prevalent pathology among infertile men. The mechanisms linking this condition to infertility, however, are poorly understood. Our previous work showed a relationship between sperm functional quality and the ability of spermatozoa to respond to capacitating conditions with increased membrane fluidity and protein tyrosine phosphorylation. Given the reported association between varicocele, oxidative stress, and sperm dysfunction, we hypothesized that spermatozoa from infertile patients with varicocele might have a combined defect at the level of membrane fluidity and protein tyrosine phosphorylation. Semen samples from infertile patients with and without grade II/III left varicocele were evaluated for motion parameters (computer-assisted semen analysis [CASA]), hyperactivation (CASA), incidence and intensity of protein tyrosine phosphorylation (phosphotyrosine immunofluorescence and western blotting), and membrane fluidity (Laurdan fluorometry), before and after a capacitating incubation (6 hr at 37 degrees C in Ham's F10/BSA, 5% CO(2)). Spermatozoa from varicocele samples presented a decreased response to the capacitating challenge, showing significantly lower motility, hyperactivation, incidence and intensity of tyrosine phosphorylation, and membrane fluidity. The findings reported in this article indicate that the sperm dysfunction associated to infertile varicocele coexists with decreased sperm plasma membrane fluidity and tyrosine phosphorylation. These deficiencies represent potential new pathophysiological mechanisms underlying varicocele-related infertility.

Colgajo miocutáneo de glúteo mayor en isla con cierre V en Y para la cobertura de úlceras isquiáticas / Island miocutaneous gluteus maximus flap for coverage of ischiatic ulcers

Las úlceras en la región isquiáticas ocurren por la permanencia en posición de sentado durante períodos prolongados. Son las úlceras con la mayor recidiva y presencia de lechos ulcerosos múltiples y sinuosos. Esto ocasiona hospitalización prolongada, aseos quirúrgicos repetidos, antibioterapia múltiple y retraso de la rehabilitación. La cobertura se puede realizar con colgajos miocutáneos de glúteo mayor, bíceps femoris, gracilis, tensor de fascia lata, semimembranoso y semitendinoso, entre otros. El colgajo miocutáneo de glúteo mayor realizado para la cobertura de úlceras isquiáticas, es actualizado en este trabajo y se incorpora como variante la utilización de una isla cutánea irrigada por perforantes, que avanza al lecho con cierre V en Y sin tensión. Nuestro objetivo es demostrar que el colgajo de glúteo mayor sería de primera elección en la cobertura de úlceras isquiáticas. Proponemos como variante quirúrgica la liberación completa del músculo, proveyendo un amplio eje de rotación, y una isla de piel sobre él, produciendo un cierre sin tensión. Todo esto reduciría la morbilidad y recidiva. Se realizaron 13 colgajos miocutáneos de glúteo mayor de avance y rotación con cierre V en Y para la cobertura de úlceras isquiáticas de presión grado III y IV. Las patologías asociadas fueron tetraplejia y paraplejia por lesión medular y paraplejia espástica familiar. El seguimiento fue de 6 meses hasta 7 años. Hasta la fecha no se ha documentado morbilidad ni recidiva. En nuestra experiencia el colgajo miocutáneo de glúteo mayor seria de elección para cierre de úlceras isquiáticas por su volumen de relleno e importante irrigación. El cierre sin tensión estaría dado por la liberación completa del músculo produciendo un amplio eje de rotación, y por una isla de piel que se transporta sobre él (AU)

The ischiatic ulcers develop as a consecuense of long sitting position. This kind of ulcers relapse in a great percentage and are a challenge for clinicians. Most of the time causes long hospital stays, multiple surgical procedures and complex antimicrobial therapy. Coverage can be done with several myocutaneous flaps, including gluteus maximus, biceps femoris, gracilis, tensor fascia lata, semitendinosus (AU)

An azoospermic man with a double-strand DNA break-processing deficiency in the spermatocyte nuclei: case report.

BACKGROUND: The mechanisms of meiotic arrest in human spermatogenesis are poorly known. METHODS AND RESULTS: A testicular biopsy from an azoospermic male showed complete spermatogenesis arrest at the spermatocyte stage, asynapsis, lack of formation of the XY body, partial reversion to a mitotic-like division and cell degeneration both at the prophase and at the abnormal cell divisions. Synaptonemal complex analysis showed minor segments of synapsis and mainly single axes. Fluorescent immunolocalization of meiotic proteins showed normal SYCP3, scarcity of SYCP1, null MLH1 foci, about 10 patches of gamma-H2AX, abnormal presence of BRCA1 among autosomal axes, absence of RAD51 in early and advanced spermatocytes and permanence of gamma-H2AX labelling up to the abnormal spermatocyte divisions that are the most advanced stage reached. There are at least six dominions of evenly packed chromatin resembling that of the normal XY body, but no true XY body. CONCLUSIONS: The protein phenotype and the fine structure of the nuclei are compatible with a deficiency of the processing of double-strand DNA breaks in the zygotene-like spermatocytes, but the features of this defect do not agree with Spo11, Sycp1, Atm and Dmc1 null mutations, which give absence of XY body, synapsis disturbances and spermatocyte apoptosis in mice.

Abnormal assembly of annulate lamellae and nuclear pore complexes coincides with fertilization arrest at the pronuclear stage of human zygotic development.

BACKGROUND: The assembly of nuclear pore complexes (NPC) and their cytoplasmic stacks, annulate lamellae (AL), promote normal nucleocytoplasmic trafficking and accompany pronuclear development within the mammalian zygote. Previous studies showed that a percentage of human oocytes fertilized in vitro failed to develop normal pronuclei and cleave within 40-48 h post insemination. We hypothesized that an aberrant recruitment of NPC proteins, nucleoporins and/or NPC preassembled into AL, might accompany human fertilization arrest. METHODS AND RESULTS: We explored NPC and AL assembly in unfertilized human oocytes, and fertilized and arrested zygotes by immunofluorescence with an NPC- and AL-specific antibody, mAb 414, and by transmission electron microscopy. Major NPC or AL assembly was not observed in the unfertilized human oocytes. Once fertilization took place, the formation of AL was observed throughout the cytoplasm and near the developing pronuclei with NPC. On the contrary, NPC assembly was disrupted in the arrested zygotes, whereas AL were clustered into large sheaths. This was accompanied by the lack of NPC incorporation into the nuclear envelopes. CONCLUSIONS: We conclude that the aberrant assembly of NPC and AL coincides with early developmental failure in humans.

A pathology of the sperm centriole responsible for defective sperm aster formation, syngamy and cleavage.

BACKGROUND: In the present report we analyse the structural and functional features of sperm from a patient with severe asthenoteratozoospermia and failure of cleavage after ICSI. METHODS: Sperm were studied by phase contrast and transmission electron microscopy and microinjected into bovine oocytes to examine aster formation using antibodies against acetylated alpha- and beta-tubulins. RESULTS: Acephalic sperm, headless tails and abnormal alignments of the head-tail junction were observed. Flagella evidenced the features of dysplasia of the fibrous sheath. Bovine oocytes injected with patient's sperm showed male and female pronuclei but a faulty development of microtubules from the sperm-derived centrosome. The first ICSI attempt using conventional sperm selection methods resulted in fertilized two pronuclei zygotes, but no syngamy or cleavage. Three more ICSI attempts were performed, carefully avoiding sperm with obvious anomalies of the connecting piece. Fertilization and cleavage took place in all cycles, and in two of them positive betahCG plasma levels were detected but preclinical abortions ensued. CONCLUSIONS: We propose that the alterations in the head-tail junction and attachment, responsible for the observed sperm phenotype, result from centriolar dysfunctions that cause insufficient sperm aster formation, lack of syngamy and cleavage or defective embryos leading to early abortions.

Sperm ubiquitination in patients with dysplasia of the fibrous sheath.

BACKGROUND: Human sperm with structural abnormalities display an increased content of the cellular proteolytic marker peptide, ubiquitin. We investigated whether dysplasia of the fibrous sheath (DFS), a severe structural anomaly found in the sperm of some asthenozoospermic patients, is accompanied by (i) increased ubiquitination of the sperm surface and (ii) by increased ubiquitination of the sperm mitochondria. METHODS AND RESULTS: Five DFS patients and eight fertile donors were studied by immunocytochemistry with anti-ubiquitin antibodies. Increased cross-reactivity of the ubiquitinated mitochondrial epitopes was seen in 32-50% of DFS sperm, but only 2-4.1% of sperm from fertile donors. Sperm surface ubiquitination assessed by sperm-ubiquitin tag immunoassay (SUTI) and immunofluorescence demonstrated an increased sperm ubiquitination in all DFS patients. The average median value of ubiquitin-induced fluorescence in DFS patients was 25.8 counts (range 19.8-37.9), as opposed to 13.4 counts range (9.3-16.6) in fertile men. Sperm with 'stump tails', coiled tails, twin and triplet sperm, and clusters of immature spermatogenic cells were common. CONCLUSIONS: DFS sperm have increased cross-reactivity to anti-ubiquitin antibodies, a finding consistent with the ubiquitination of defective sperm shown in animal models. These results justify the use of ubiquitin-based assays for objective semen analysis in infertile men with heritable defects.

Pronuclear abnormalities and cytoskeletal organization during assisted fertilization in a patient with multifollicular ovarian response.

PURPOSE: To analyze the distribution of a tubulins and acetylated alpha tubulins and the chromatin configuration in abnormally fertilized zygotes from a patient with a multifollicular ovarian response after in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI). METHODS: Immunofluorescence and phase contrast microscopy was performed in abnormally fertilized zygotes. RESULTS: After phase contrast microscopy analysis, immunofluorescence staining was performed in 20 oocytes that developed > or = 3 pronuclei (PN) and karyomeres after IVF-ICSI. Around 80% of the abnormal zygotes from IVF were the consequence of monospermic fertilizations. Retention of the second polar body (PB) and the presumptive split of > or =1 PN within the cytoplasm were the main events present in most oocytes after IVF-ICSI. CONCLUSIONS: Fluorescence labeling of selected sperm and oocyte components affords a unique view of abnormal fertilized zygotes. Surprisingly, anomalies detected after IVF-ICSI showed similar etiologies in this special group of zygotes.

Male antisperm antibodies: association with a modified sperm stress test and lipid peroxidation.

We previously reported a modified sperm stress test (MOST), low scores (< 0.39) in which were associated with sperm-related abnormal in vitro fertilization. Preliminary observations suggested that the presence of male sperm antibodies (ASA) could give low MOST scores. It was therefore decided to undertake a study to verify this possible association and also to ascertain if such a relationship was causal in nature. Six hundred and fifty semen samples from patients consulting for infertility were assessed for basic seminal characteristics, motion parameters (CASA), ASA and MOST. Thirty-nine samples (6%) were ASA-positive. Samples with and without ASA showed similar characteristics, except for percentage of normal forms and MOST scores (0.35 +/- 0.03 vs. 0.67 +/- 0.01, P < 0.001, for ASA-positive and -negative, respectively). There was a strong statistical association between presence of ASA and low MOST scores (P < 0.0001). One-hundred per cent of ASA-positive samples displayed low MOST scores. To verify the nature of this relationship, we incubated ASA-free spermatozoa with ASA-positive and -negative (control) sera. Despite an increase in the percentage of ASA-bearing spermatozoa in those aliquots incubated with ASA-positive serum, their original (pre-incubation) MOST scores remained unchanged. Furthermore, the rate of lipid peroxidation, indirectly reflected in MOST scores, was not different in the aliquots incubated with ASA. In conclusion, there seems to be a strong association between presence of ASA and low MOST values in semen samples of infertile patients; however, the relationship does not appear to be causal.

Serum inhibin B may be a reliable marker of the presence of testicular spermatozoa in patients with nonobstructive azoospermia.

OBJECTIVE: To establish the predictive value of serum inhibin B levels as an indicator of the presence of testicular spermatozoa in nonobstructive azoospermia, compared with the traditional serum FSH marker. DESIGN: Prospective study. SETTING: Private high-complexity reproductive center with university affiliation. PATIENT(S): Seventy-eight patients with nonobstructive azoospermia, 15 patients with obstructive azoospermia, and 10 fertile volunteers. INTERVENTION(S): Blood samples, testicular sperm extraction, percutaneous epididymal sperm aspiration, and semen collection. MAIN OUTCOME MEASURE(S): Serum levels of inhibin B and FSH and presence of spermatozoa on TESE, PESA, or regular semen analysis. RESULT(S): Patients with nonobstructive azoospermia has significantly higher levels of serum FSH and significantly lower levels of inhibin B. Mean inhibin B serum levels were significantly higher in patients with nonobstructive azoospermia who had spermatozoa on TESE than in those in whom no spermatozoa were found (89.31 +/- 73.24 pg/mL vs. 19.23 +/- 22.34 pg/mL), but mean FSH serum levels did not have similar predictive power (21.37 +/- 12.92 IU/mL vs. 19.27 +/- 10.28 IU/mL). The cut-off level of inhibin B separating both groups, as determined by the receiver-operating characteristic curves, was >53 pg/mL. CONCLUSION(S): Serum inhibin B level seems to be more accurate than serum FSH level in prediction of the presence of testicular spermatozoa in patients with nonobstructive azoospermia.

Incidence of tail structure distortions associated with dysplasia of the fibrous sheath in human spermatozoa.

Dysplasia of the fibrous sheath (DFS) is an anomaly found in spermatozoa of severe asthenozoospermic patients. Marked hypertrophy and hyperplasia of the fibrous sheath is the common characteristic. Immunocytochemistry allowed us to visualize the distortions and incidence of tail structure abnormalities associated with this phenotype in six patients; four with a complete form and two with an incomplete form of this pathology previously diagnosed and studied by electron microscopy. Microtubules and fibrous sheaths were studied using monoclonal antibodies against alpha-acetylated tubulin and anti-FSC1 (the major protein component of the fibrous sheath). Mitochondrial sheaths were visualized using the mitochondrion-specific vital dye MitoTracker green FM(TM). Phase contrast and fluorescent microscopy of semen samples showed large numbers of spermatozoa with short, rigid, thick and irregular tails. As expected, anomalous and completely distorted fibrous sheaths, severe alterations of the axonemal microtubules and different patterns of mitochondrial sheath configurations were found. While ultrastructural studies of thin sections allow an in-depth knowledge of the internal organization of the sperm tail, fluorescence labelling of selected sperm components affords a unique view of the whole flagellum including topographical relationships of various organelles. The combination of these different approaches is essential for a comprehensive understanding of this particular pathology.

Conservative management versus aspiration of functional ovarian cysts before ovarian stimulation for assisted reproduction.

PURPOSE: To compare the efficiency of transvaginal ultrasound-guided functional ovarian cyst aspiration, with conservative management, in the outcome of patients undergoing assisted reproductive technique (ART) (in vitro fertilization or intracytoplasmic sperm injection). These cysts were identified before ovarian stimulation begun and after administration of a midluteal GnRH agonist. METHODS: Fifty nine patients undergoing ART from January 1, 1997 to February 28, 1999, who developed functional ovarian cysts were included. Aspirations of these cysts (n = 14) versus conservative management (observation) (n = 45) were compared. Total number of ovarian follicles developed, number of oocytes retrieved, estradiol levels on the day of human chorionic gonadotropin, fertilization rate, number of good quality embryos transferred, implantation, and clinical pregnancy rate per cycle were evaluated. RESULTS: No statistical differences were observed between the two groups in any of the selected parameters. CONCLUSIONS: Cyst aspiration and conservative management showed similar implantation and pregnancy rates, in patients who develop functional ovarian cysts after pituitary down-regulation following luteal phase gonadotropin-releasing hormone agonist administration. Prospective studies are needed to confirm this trend.

Cytoskeletal organization defects and abortive activation in human oocytes after IVF and ICSI failure.

In this study, we analysed the distribution of beta tubulins to detect spindle and cytoplasmic microtubules, alpha acetylated tubulins for sperm microtubules and chromatin configuration in oocytes showing fertilization failure after conventional IVF or intracytoplasmic sperm injection (ICSI). A total of 450 human oocytes that failed to fertilize were studied 20-40 h after IVF or ICSI. In all, 287 oocytes were stained for immunofluorescence and chromosomal spreads were performed by Tarkowski's air-drying method in 163 IVF or ICSI oocytes that did not develop pronuclei after the extrusion of a second polar body. Immunofluorescence analysis showed that the main reason of fertilization failure after IVF was no sperm penetration (55.5%). The remaining oocytes showed different abnormal patterns, e.g. oocyte activation failure (15.1%) and defects in pronuclei apposition (19.2%). On the other hand, fertilization failure after ICSI was mainly associated to incomplete oocyte activation (39.9%), and to a lesser extent with defects in pronuclei apposition (22.6%) and failure of sperm penetration (13.3%). A further 13.3% of the ICSI oocytes arrested their development at the metaphase of the first mitotic division. The chromosomal spreads allowed the analysis of abortive activations, in which no pronuclei formed but a second polar body was extruded. Immunofluorescence and cytogenetic analysis provided a useful tool to improve infertility diagnosis and prognosis in each particular case.

Pregnancies established through intracytoplasmic sperm injection (ICSI) using spermatozoa with dysplasia of fibrous sheath.

AIM: Dysplasia of the fibrous sheath (DFS) is an anomaly found in asthenozoospermic patients with extremely low or absent motility. In order to determine the efficacy of ICSI in these patients, a retrospective analysis of ICSI results in DFS patients has been done. METHODS: Ten ICSI attempts were performed in 6 patients with diagnosis of Dysplasia of the Fibrous Sheath studied by transmission and scanning electron microscopy. RESULTS: In the cases studied, sperm concentration was (29.62 +/- 18.05) x 10(6)/mL, total motility was 1.14 +/- 1.31%. Progressive motility was 0% except for one case with 0.1% . One hundred and three preovulatory oocytes were obtained and 94 metaphase II oocytes were injected. Sixty-nine of them showed two pronuclei (fertilization rate: 73.4%). Forty-nine embryos were obtained and 34 were transferred (mean: 3.4 embryos per transfer). Five pregnancies were diagnosed by beta-hCG plasma level determinations that resulted to be one preclinical abortion, one clinical abortion and three deliveries. Another pregnancy (ongoing) was achieved from a cryopreserved embryo transfer. CONCLUSION: These results showed that ICSI provides a suitable solution for patients suffering from irreversible sperm defects such as DFS. Nevertheless, it is mandatory to inform couples of possible transmission risks to offspring, which are unknown at present. Only when the etiology of this problem is disclosed, it will be possible to assess the real genetic risk.

Acephalic spermatozoa and abnormal development of the head-neck attachment: a human syndrome of genetic origin.

A series of 10 young sterile men with acephalic spermatozoa or abnormal head-mid-piece attachments is presented. Nine of these patients had 75-100% spermatozoa with minute cephalic ends and 0-25% abnormal head-middle piece attachments. Loose heads ranged between 0-35 for each 100 spermatozoa and normal forms were rare. Two patients were brothers. On ultrastructural examination, the head was generally absent and the middle piece was covered by the plasma membrane. When present, heads implanted at abnormal angles on the middle piece. A testicular biopsy showed abnormal spermiogenesis. The implantation fossa was absent and the flagellar anlage developed independently from the nucleus, resulting in abnormal head-middle piece connections. In one patient azoospermia was induced with testosterone to attempt to increase the normal sperm clone during the rebound phenomenon, but all newly formed spermatozoa were acephalic. In another patient with high numbers of defective head-mid-piece connections, microinjections of spermatozoa resulted in four fertilized oocytes, but syngamy and cleavage did not take place, suggesting an abnormal function of the centrioles. The findings indicate that acephalic spermatozoa arise in the testis as the result of an abnormal neck development during spermiogenesis. The familial incidence and the typical phenotype strongly suggest a genetic origin of the syndrome.