A new method to process testicular sperm: combining enzymatic digestion,accumulation of spermatozoa,and stimulation of motility

Purpose

In azoospermia processing of the TESE material often results in a sample of reduced purity. This prospective study was set up to clarify whether a combination of enzymatic digestion, density gradient centrifugation and stimulation of motility (where indicated) is a feasible option in TESE patients.

Methods

A total of 63 samples (showing spermatozoa) were processed by the present tripartite processing method. The resulting sperm sample of high purity was directly used for ICSI and subsequent cryopreservation when quality of the accumulated sperm sample allowed for it (n = 39 cycles).

Results

Compared to the control group blastocyst formation rate in the present tripartite processing technique was significantly (P < 0.01) higher (55.2 vs. 43.7 %). Fertilization rates differed significantly (P < 0.001) between cases in which motile sperm could be used (58.4 %) compared to ICSI with immotile sperm (45.0 %). Clinical pregnancy rate per transfer was 40.0 % (24/60) using fresh and 21.6 % (8/37) with cryopreserved TESE material. The calculated live birth rates were 31.7 and 21.6 %, respectively. Thirty-five healthy children were born.

Conclusions

A comparison with a control group suggests that the present approach using standardized ready-to-use products is efficient and reliable. Presumably healthy live births further indicate the safety of the procedure.

     

Vascular endothelial growth factor and its receptors in male fertility.

OBJECTIVE: To determine the concentration of vascular endothelial growth factor in the seminal fluid, the presence of vascular endothelial growth factor receptors Flt-1 and KDR in spermatozoa, and the predictive value of seminal vascular endothelial growth factor on fertilization and the chance of pregnancy in patients undergoing intracytoplasmic sperm injection (ICSI) or IVF. DESIGN: Retrospective analysis. SETTING: Private institute (semen collection, IVF/ICSI) and academic research environment (analysis of seminal fluid and spermatozoa). PATIENT(S): Eighty men whose spermatozoa were subsequently used for IVF or ICSI. INTERVENTION(S): Seminal vascular endothelial growth factor was measured by an EIA. Spermatozoa were analyzed by fluorescence-activated cell sorter analysis and by immunocytochemistry. MAIN OUTCOME MEASURE(S): Oocyte fertilization rate, pregnancy rate, and presence of vascular endothelial growth factor receptors on spermatozoa. RESULT(S): Patients with a seminal concentration of vascular endothelial growth factor of 2-100 ng/mL had a sixfold increased chance of pregnancy. Vascular endothelial growth factor concentration and patient's age remained the only independent prognostic factors for pregnancy. The concentration of vascular endothelial growth factor did not correlate with indices of male factor infertility or with the oocyte fertilization rate. Expression of vascular endothelial growth factor receptors (Flt-1, KDR) on spermatozoa was demonstrated. CONCLUSION(S): The seminal concentration of vascular endothelial growth factor correlates with the chance of pregnancy in patients undergoing IVF or ICSI. Vascular endothelial growth factor receptors Flt-1 and KDR were detected on spermatozoa for the first time.     

Axillary Metastases of an Occult Primary Carcinoma of the Breast—Discovered Only byTc-Tetrofosmin Scintigraphy

Recent reports consider^99mTc-tetrofosmin scintigraphy to be a powerful new diagnostic tool for discriminating malignant from benign breast disease. We report on a woman suffering from histologically confirmed axillary metastases of a primary unknown, occult carcinoma, whose origin was suspected within the breast. All the diagnostic procedures performed to discover any lesion failed or were inconclusive. The primary cancer was clearly visualized, however, in the right breast by means of^99mTc-tetrofosmin scintigraphy.Conclusion.We suggest that^99mTc-tetrofosmin scintigraphy is a powerful method to detect breast cancer, especially when other diagnostic imaging procedures are inconclusive.     

Endocrinology: Prediction of ovarian hyperstimulation syndrome of baseline ovarian volume prior to stimulation

The aim of the study was to find out whether the estimationof the baseline ovarian volume prior to stimulation would bea suitable predictor for the risk of ovarian hyperstimulationsyndrome (OHSS). A total of 101 patients underwent in-vitrofertilization (IVF) and embryo transfer. They had a 3-D volumetricassessment of the ovaries and body weight estimations on thefirst day of hormonal stimulation. A second measurement wasperformed on the day of ovulation induction with human chorionicgonadotrophin (HCG) together with an oestradlol 17 estimationin serum. During the IVF programme 15 women developed OHSS and86 did not. There was a significant correlation between thebaseline ovarian volume and subsequent occurrence of OHSS (P=0.03).Other significant relationships were foimd between the occurrenceof OHSS and the number of follicles (P=0.002), the number ofoocytes retrieved (P=0.0001) and the length of the cycle (P=0.0001).The body weight before and after the stimulation was significantlylower in the group of women who did develop the syndrome (P=0.011resp.0.03). The oestradiol 17 concentration on the day of HCGadministration in the serum of the patients who had OHSS wassignificantly higher (P=0.0001). In conclusion, voluinetry ofthe ovaries could help to detect patients at risk and preventthe occurrence of OHSS by early adjustment of the hormonal dosage.Recent advances in ultrasound technology (3-D ultrasound) enablequick and highly accurate volumetric assessments. Furthermore,our study confirms previous observations that low body weightand long cycles seem to be additional risk factors for the developmentof OHSS.     

Fertilization and early embryology: Use of lasers in assisted fertilization and hatching

The erbium-yttrium-aluminium-garnet (Er: YAG) laser has beenapplied to micromanipulation in humans. It was used in the fertilizationprocess for both subzonal insemination (SUZI) and for partialzona dissection (PZD). Laser-assisted micromanipulation achievedsignificantly higher fertilization rates (34.8%) when comparedto mechanical SUZI (16.1%), but use of the laser did not improvethe PZD results (laser 14.8% versus mechanical 14%). The Er:YAG laser was used to assist hatching. In the mouse it significantlyimproved the hatching rate (80 versus 29.3%) 110 h after administrationof human chorionic gonadotrophin. This technique was appliedin two different centres to patients with previous in-vitrofertilization (IVF) failures. The implantation rate per embryo(14.4% laser-assisted hatching versus 6% control group) andthe pregnancy rate per transfer (40 versus 16.2%) were improved.     

Number and Size of Antral Follicles as Predictive Factors in In Vitro Fertilization and Embryo Transfer

Purpose: The purpose of the study was to evaluate whethernumber and size of antral follicles can predict the outcomeof in vitro fertilization—embryo transfer. Methods: A total of 113 patients were prospectively includedinto this study. After 19 days of down-regulation, numberand size of follicles were determined by using recent three-dimensionaltransvaginal ultrasound technology. Beforeapplication of gonadotropin, all follicles had been definedas antral follicles. According to size, antral follicles werecategorized into four different groups: group I includedantral follicles < 5 mm, group II follicles 5–10 mm; groupIII 11–20 mm; and group IV >20 mm. Pregnant and non-pregnantpatients were compared in terms of their numberof antral follicles of group I–IV. These four groups were thencompared regarding implantation rate, number of retrievedoocytes, endometrium thickness, and age. Results: Pregnant patients showed an significant highernumber of follicles with the size between 5 and 10 mm (P=0.04). A significant correlation was found between numberof retrieved oocytes and antral follicle size of 5–10 mm(P= 0.0001). Antral follicles with a diameter between 5and 10 mm decreased significantly with age (P = 0.008).In groups III and IV, a significant correlation was foundbetween antral follicle size (P = 0.016) and serum estradiollevel after gonadotropin-releasing hormone-agonist down-regulation (P = 0.011). Conclusions: We demonstrated that patients with a highernumber of follicles between 5 and 10 mm showed a significantlyhigher pregnancy rate, whereas patients with a dominantnumber of antral follicles > 11 mm have a highercancellation rate due to ovarian low response.