Assisted reproduction in male cancer survivors: fertility treatment and outcome in 67 couples

BACKGROUND: Many male cancer survivors experience fertility problems due to antineoplastic treatment. We report the fertility outcome in 67 couples referred to assisted reproduction treatment (ART) because of male factor infertility due to cancer. METHODS: This was a retrospective study assessing the following parameters: diagnosis, cancer treatment, type of fertility treatment and type of sperm used, number of pregnancies and pregnancy outcome. RESULTS: Testicular cancer and lymphomas were the most prevalent diagnoses. Adjuvant treatment with chemo- and/or radiation therapy had been given to 90% of the men. Semen was cryopreserved in 82% of the men prior to treatment. Following antineoplastic treatment, 43% of the men had motile spermatozoa in the ejaculate, but 57% were azoospermic. A total of 151 ART cycles were performed [55 intra-uterine insemination (IUI), 82 ICSI and 14 ICSI-frozen embryo replacement (FER)]. The clinical pregnancy rate per cycle was 14.8% after IUI, 38.6% after ICSI and 25% after ICSI-FER. The corresponding delivery rates were 11.1, 30.5 and 21%. Cryopreserved semen was used in 58% of the pregnancies. The delivery rate per cycle was similar after use of fresh or cryopreserved spermatozoa. CONCLUSIONS: Male cancer survivors have a good chance of fathering a child by using either fresh ejaculated sperm or cryopreserved sperm.     

Simple,noninvasive system for measuring the heart rate of avian embryos and hatchlings by means of a piezoelectric film

Using a flexible piezoelectric film, the authors developed a simple system to determine noninvasively the heart rate of chicken embryos and hatchlings. The film was piezoelectric polyvinylidene fluoride (PVDF), which is sensitive enough to detect cardiogenic ballistic movements of the egg (ballistocardiogram (BCG)) and precordial movements of the hatchling attributable to cardiac contractions (apexcardiogram (ACG)). The BCG could be detected, during the second half of incubation, by placing the egg on the PVDF film on a soft substrate. The detected signal was found to be a measure of movement velocity. The ACG could be measured when the hatchling's chest wall made contact with the PVDF film installed in a box in which the hatchling was confined. The heart rate was counted from the lag time of autocorrelation calculated for a certain time segment (e.g. 2s) of the BCG and ACG recordings.     

Cryopreservation of human mononuclear cells for quality control in clinical immunology. I. Correlations in recovery of K- and NK-cell functions,surface markers,and morphology

Cryopreserved human peripheral blood mononuclear cells (PBMC) were tested for natural killer (NK) and antibody-dependent cellular cytotoxicity (ADCC) and for high-affinity (29°C) and total (4°C) rosette formation with sheep erythrocytes. PBMC produced variable NK activity following freezing and thawing, but consistently reacted well in ADCC. A significant correlation was found between low NK activity and a decreased percentage of low-affinity rosette-forming cells. On the contrary, the number of large granular lymphocytes (LGL), among which NK cells are restricted, and the reactivity with the monoclonal antibody OKT10, which recognizes the majority of LGL in the peripheral blood, were not significantly altered by cryopreservation. Cryopreserved cells proved to be excellent controls for determining the day-to-day variability of the NK assay and for selecting optimum conditions for this test in the clinical immunology laboratory.     

Fluorescence in situ hybridization analysis of two blastomeres from day 3 frozen-thawed embryos followed by analysis of the remaining embryo on day 5

BACKGROUND: Chromosomal mosaicism in human embryos may give rise to false positive or false negative results in preimplantation genetic diagnosis for aneuploidy screening (PGD-AS). Therefore, we have investigated whether the results obtained from a 2-cell biopsy of frozen-thawed embryos and fluorescence in situ hybridization (FISH) analysis are representative for the chromosome constitution of the remaining embryo on day 5. METHODS: Cryopreserved day 3 embryos were thawed and from surviving embryos two blastomeres were biopsied. FISH analysis was performed for chromosomes 1, 7, 13, 15, 16, 18, 21, 22, X and Y. After biopsy, the embryos were cultured until day 5 and further analysed using the same probe panels. RESULTS: In all, 17 embryos were available with a diagnosis based on two blastomeres on day 3 and confirmatory studies on day 5. In 10 of these 17 cases the initial diagnosis could be confirmed. However, in only six cases cytogenetic results were concordant. Besides the 10 cases with a 'correct' diagnosis, there were six false positive results and one false negative, all involving mosaicism. CONCLUSIONS: Investigating the chromosomal constitution of two blastomere nuclei offers a good opportunity to study the incidence of chromosomal mosaicism in early embryo development. The confirmation rate of the results obtained on day 3 depends on the interpretation and is higher when considered from a clinical than from a cytogenetic point of view.     

胰岛素、可地松和已烯雌酚处理妊娠早期小鼠对其胚胎发育的影响

本文用不同剂量的胰岛素、可地松、已烯雌酚,作用于妊娠开始至80小时的昆明小鼠,然后观察胚胎早期(植入前)和胚胎晚期(妊娠18天)的胚胎数量、发育时期及其与黄体数相比成活率的变化。实验表明,用较大剂量激素处理后,早期胚胎的上述各项指标,均表现出十分明显的抑制效果。当剂量逐渐降低后,仍表现出不同程度的抑制作用,只是逐渐趋于正常。所用各种激素对晚期胚胎的影响,表现在除部分死亡外,存活者的生长和发育一直落后。说明妊娠早期小鼠体内某些激素的水平,不仅直接影响早期,而且也影响晚期胚胎的发育。     

细胞因子调节对纤连蛋白在孕早期小鼠子宫内膜表达的研究

目的:通过研究细胞因子对FN表达的调节,探讨FN在围着床期子宫内膜表达的调节机理。方法:给孕早期小鼠注射不同的细胞因子,收集子宫内膜。分别用Immuno-blot法及RT-PCR法测定子宫内膜FN蛋白和nRNA的水平。结果:LIF低、高剂量组的FNmRNA和蛋白水平都比对照组高,统计分析差异有显著性（P<0.05,P<0.01）,IL-1和IL-1ra各组FN水平与对照组比无显著差异。结论:LIF在转录和转录后水平上调早期子宫内膜FN的表达,LIF参与着床与调节粘附分子的表达有关。IL-1系统则不影响子宫内膜FN的合成。     

The value of sperm pooling and cryopreservation in patients with transient azoospermia or severe oligoasthenoteratozoospermia.

BACKGROUND: A transient state of azoospermia may occur due to toxic, environmental, infectious or iatrogenic conditions. Finding sperm in the ejaculate of such patients is often unpredictable and may be critical in IVF treatment. In the present study, the approach of pooling and cryopreservation of sperm is evaluated. Cryopreservation was performed in a unique group of patients in whom no sperm had been found in at least one previous sperm examination and in patients diagnosed as suffering from non-obstructive azoospermia in whom, occasionally, sperm were found. METHODS: A total of 157 semen pooling and cryopreservation procedures in 53 patients was performed between January 1998 and December 2000 in our centre. Forty five of these patients underwent an IVF-ICSI treatment during the study period. In 32 patients, fresh sperm were used to perform ICSI. In 13 patients no sperm were available, and the previously frozen sperm were used. RESULTS: Using our pooling system, 13 IVF-ICSI cycles were rescued. In seven patients with a previous testicular biopsy due to azoospermia, sperm cryopreservation was possible. Overall, 13 pregnancies (10 deliveries, two ongoing pregnancies and one missed abortion) were achieved. CONCLUSION: The introduction of semen banking for patients with transient azoospermia may increase the chance of pregnancy using their own sperm.     

Pregnancies after intracytoplasmic sperm injection with cryopreserved testicular spermatozoa

In 25 patients (14 suffering from obstructive azoospermia, sixfrom non-obstructive azoospermia, three from astheno-azoospermiaand two from absence of ejaculation) spermatozoa were extractedfrom testicular biopsies. Intracytoplasmic sperm injection (ICSI)with fresh testicular spermatozoa was performed in 18 cases;spermatozoa in excess were cryopreserved in pills. No pregnancieswere achieved. In the remaining seven patients, testicular spermatozoawere retrieved and cryopreserved during a diagnostic testicularbiopsy. After thawing, sperm motility was assessed in 17 cases(68%), and 18 ICSI with cryopreserved testicular spermatozoawere performed. The mean two-pronuclear (2PN) fertilizationrate was 59%, the mean cleavage rate was 92%, and six clinicalpregnancies were achieved, all of them still ongoing (pregnancyrate 33%). A comparison of the results of ICSI carried out withfresh or cryopreserved testicular spermatozoa showed that themean 2PN fertilization rates per cycle (53 compared with 55%),mean cleavage rates per cycle (99 compared with 96%) and embryoquality were not significantly different In conclusion, cryopreservationof testicular spermatozoa is feasible, even in patients withnon-obstructive azoospermia, and the results of ICSI with frozen-thawedtesticular spermatozoa are similar to those obtained using freshtesticular spermatozoa. Cryopreservation of testicular spermatozoamay avoid repetition of testicular biopsies to retrieve spermatozoafor successive ICSI cycles in patients in whom the only sourceof motile spermatozoa is the testicle.     

The influence of cooling on the oranization of meiotic spindle of the mouse oocyte

The effect of cooling on the organization of the microtubulesystem of the mouse oocyte has been investigated. Cooling to25, 18 or 4°C for varying periods of time resulted in aprogressive disassembly of the spindle and the dispersal ofthe chromosomes. The extent of the changes observed was greaterat lower temperatures and with longer periods of exposure. Transferof oocytes from either 37 or 4 to 24°C resulted in the rapidand transient appearance of polar asters and of multiple cytoplasmicasters associated with the pericentriolar material present atthe spindle poles and in the cytocortex of the oocyte. Thistransient aster formation appeared to be driven by the elevatedlevels of free tubulin released during spindle disassembly.An apparent reversal of many of the changes induced by low temperaturewas observed in many but not all oocytes on their restorationto 37°C for 1 h. These results have implications for ourunderstanding of microtubule organization in the oocyte, andfor the handling of oocytes during IVF and GIFT therapeuticprocedures and during the cryopreservation of oocytes.