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The inability of sperm chromatin to decondense has been implicated in the failure of fertilization, This study was undertaken to identify the relationship between sperm chromatin decondensation in vitro after incubation with follicular fluid at various points in time and fertilization or pregnancy rates after intracytoplasmic sperm injection. Moreover, an attempt was made to determine whether this test could be used as a predictive test for the outcome of ICSI. Thirty-two infertile couples undergoing ICSI therapy were included in this prospective study. One milliter of semen from each sample was mixed with 1 mL of follicular fluid obtained from ICSI patients at the time of oocyte retrieval and incubated for 24 h. Many smears were made directly after semen liquefaction at the following time intervals: 30, 60, and 120 min and 24 h. Chromatin decondensation was evaluated with acridine orange staining. The mean percentage of uncondensed chromatin of spermatozoa in the native semen samples was 25 ±18.3%, which increased within 24 h to 91 ±9.5%. On the other hand, the fertilization and ongoing pregnancy rates were 64 ±21.7% and 20%, respectively. However, no correlations were found between chromatin decondensation at various point of time (30, 60, and 120 min and 24 h) and fertilization rate. No correlation was shown between the chromatin decondensation and sperm counts in the ejaculate, morphology, or the percentage of condensed chromatin. In light of this study, chromatin decondensation in vitro cannot be recommended for predicting the fertilization potential of spermatozoa and pregnancy rates in the ICSI program. Further research is necessary, especially in cases where ICSI is being considered as a therapeutic option.

Semen analysis constitutes the most important investigation of male infertility. However, the true anomalies present in defective sperm cells have been only partially characterized. The integrity of the sperm chromatin may play the most important role, particularly in ICSI, where most of the natural selection mechanisms are bypassed. This study was carried out to characterize sperm morphology (strict criteria), to evaluate chromatin condensation and sperm count in native semen as well as after semen preparation by the swim-up technique, and to eventually evaluate any correlation between these parameters. Semen from 90 men was analyzed for the above parameters in both the fresh and processed semen. Whereas the sperm count decreased after sperm preparation by the swim-up technique in comparison to the value in the fresh semen (p<.001), there was an increase in the percentage of morphologically normal (p<.001) and chromatin-condensed sperm (p=.99). However, there was no correlation between sperm morphology, chromatin condensation, and sperm count either in the fresh or in the processed semen samples. These results suggest that sperm morphology, sperm count, and chromatin condensation are independent parameters that should be evaluated separately in the assessment of male fertility in an assisted reproduction program.

Abstract: Anaesthetic management for oocyte retrieval may interfere with the results and success rate of an in vitro fertilization (IVF) program. This study was conducted to compare the effects of two different anaesthetic techniques (general anaesthesia versus sedation) used for oocyte retrieval on IVF outcome. For general anaesthesia we used a combination of remifentanil (UltivaTM) with either propofol or isoflurane in hypnotic concentrations. For sedation the protocol included midazolam, diazepam or propofol according to clinical needs. In total, 202 women were enrolled in the study. 96 women opted for sedation and 106 for general anaesthesia. The number of collected oocytes was significantly higher with general anaesthesia (10.54±5.43 [mean±SD]) than with sedation (6.25±3.65, p<0.0001), whereas the number of fertilized oocytes was not different (4.70±3.57 vs. 4.23±2.90). There were no significant differences in cleavage and pregnancy rates. We therefore believe that remifentanil-based general anaesthesia without nitrous oxide is a suitable alternative to sedation and may be recommended for IVF oocyte retrieval if general anaesthesia is requested.

A significant association between male subfertility, imperfect spermiation and abnormal nuclear condensation has been suggested. The DNA content of spermatozoa might be responsible for inducing alterations in sperm morphology. The final nuclear shape, which is species-specific, depends on chromatin condensation during spermatogenesis as well as a precise organization of DNA within the nucleus. Many reports have described the association between disturbances in sperm chromatin condensation, morphology and male infertility. Chromatin condensation is achieved by gradual substitution of lysinerich somatic histones by testis-specific histone and finally by protamine. In this study two groups of patients were compared: the first consisted of 63 patients who had undergone intracytoplasmic sperm injection (ICSI) with freshly ejaculated spermatozoa whereas the second included 47 patients assigned to ICSI with testes biopsy-extracted spermatozoa. In both groups chromatin condensation was assessed by aniline blue staining and morphology evaluated according to strict criteria. The condensed chromatin and morphology of spermatozoa were significantly (P < 0.0001) less in the second group compared to the first. However the fertilization, cleavage, implantation and pregnancy rates were almost the same in both investigated groups. There was no significant difference between the two groups with respect to ICSI outcome. The percentage of chromatin condensation (nuclear maturity) and morphologically-normal spermatozoa were significantly higher (P < 0.0001) in the ejaculated spermatozoa than in those from testis biopsy but the ICSI outcome (fertilization, cleavage, implantation and pregnancy rates) was the same. In view of these results the fertilization capability and the embryo quality obtained using testis biopsy extracted spermatozoa is not influenced by chromatin condensation and sperm morphology in testicular sperm extraction (TESE)-ICSI programmes. Therefore, it could be said that neither chromatin condensation nor morphology of testis extracted sperm could predict the fertilization, implantation and pregnancy rate in TESE-ICSI programmes.

Remifentanil ist ein potentes, gut steuerbares Opioidanalgetikum mit schwacher hypnotischer Eigenwirkung, das bei alleiniger Anwendung dosisabhängig eine Schmerzausschaltung ermöglichen kann, ohne daß der Patient das Bewußtsein verliert. Bei kurzen, aber schmerzhaften Eingriffen wie der transvaginalen Follikelpunktion kann dies ein Vorteil sein, wenn dadurch auf die sonst häufig übliche Allgemeinanästhesie verzichtet werden kann. Die Qualität einer solchen Infusionsanalgesie mit Remifentanil bei wachen Patienten sollte in der vorliegenden Studie untersucht werden. Methodik: Patientinnen, bei denen eine transvaginale Follikelpunktion vor In-vitro-Fertilisation (IVF) geplant war, erhielten keine Prämedikation und wurden wach in den OP gebracht. Die Remifentanil-Infusion wurde mit 0,25 µg/kg/min gestartet; sobald die Frauen z.B. über ein Schwindel- oder Wärmegefühl berichteten, konnte mit der Follikelpunktion begonnen werden. Die Infusion wurde bei Bedarf in Schritten von 0,05 µg/kg/min angepaßt und mit der letzten Punktion beendet. Alle Vitalparameter inkl. der pulsoxymetrisch gemessenen Sauerstoffsättigung (psO2) wurden 2,5minütlich erfaßt, außerdem Dosisbedarf, Nebenwirkungen, Sedierungsgrad und mittels Fragebogen die Patientenzufriedenheit. Zusätzlich wurden die Remifentanil-Plasmakonzentrationen durch pharmakokinetische Simulation (3-Kompartimentmodell, STANPUMP-Software®) ermittelt. Ergebnisse:Insgesamt wurden die Daten (x¯±SD) von 50 Frauen im Alter von 31,8±5,1 Jahren erfaßt. Remifentanil wurde bei 70% der Patientinnen durchgehend mit 0,25 µg/kg/min dosiert, in 22% war eine Dosissteigerung auf 0,3 µg/kg/min, in 2% auf 0,4 µg/kg/min erforderlich, bei 6% mußte die Dosierung auf 0,2 µg/kg/min reduziert werden. Währenddessen blieben die Patientinnen meist wach oder waren sofort erweckbar; Blutdruck, Herz- und Spontanatemfrequenz sowie die psO2waren im Mittel nahezu unverändert. Die berechneten Remifentanil-Plasmakonzentrationen betrugen bei OP-Beginn 5,0±1,3, bei OP-Ende 6,6±1,3 und bei Ankunft im AWR 1,2±0,5 ng/ml. Insgesamt berichteten 54% der Frauen über Juckreiz, eine Thoraxrigidität trat nicht auf, 94% aller Patientinnen würden diese „Anästhesieform” nochmals wählen. Schlußfolgerungen: Bei IVF-Follikelpunktionen ist die alleinige Anwendung von Remifentanil zur Operationsanalgesie eine gute Alternative zur Allgemeinanästhesie. Die Patientenzufriedenheit ist hoch, eine sorgfältige anästhesiologische Überwachung – insbesondere der Atemfunktion – muß aber jederzeit gewährleistet sein. Introduction:Transvaginal puncture for oocyte retrieval is a short-lasting but painful procedure. We hypothesized that a sole infusion of the ultra-short acting µ-agonist remifentanil may be a suitable and well-controllable single-agent analgesic technique that can dose-dependently be applied to spontaneously breathing patients. Methods: Fifty consenting adult women were enrolled in this prospective trial. A sedative premedication was omitted, all patients received 3 L/min of inhaled oxygen, and a sole remifentanil infusion was started with 0.25 µg/kg/min. Remifentanil was adjusted as needed for pain relief (in steps of 0.05 µg/kg/min) and finished after the last puncture. Dosage requirements, vital functions, oxygen saturation (as achieved by pulse oximetry, psO2), adverse drug effects and the level of sedation (LOS 1–5; 1=asleep/unarousable, 4=calm/awake) were recorded. Remifentanil plasma concentrations were achieved by STANPUMP pharmacokinetic simulation. Data are presented as mean±SD. Results:A total of 50 women (31.8±5.1 yr, 67.3±14.3 kg, ASA I or II ) were investigated. Follicular aspiration lasted 10.8±5.2 min, and remifentanil was infused for 19.7±8.3 min. Dosage requirements were 0.25 µg/kg/min in 70% of all patients, 0.3 µg/kg/min in 22%, 0.2 µg/kg/min in 6%, and 0.4 µg/kg/min in 2% of all cases. Vital signs (baseline, after 1stpuncture, end of surgery) nearly remained unchanged: heart frequency=85±15, 87±17, 90±17 bpm, systolic blood pressure=129±12, 132±13, 131±14 mmHg; respiratory rate=15±3, 16±4, 15±4 breaths/min; psO2=99±1, 99±1, 99±2%. LOS was 4.0 (all), 3.9±0.3, 3.9±0.3. Remifentanil plasma concentrations were 5.0±1.3 ng/mL at the start, 6.6±1.3 at the end of surgery and 1.2±0.5 at PACU arrival. Adverse drug effects: 54% itching, no muscle rigidity. 94% of all women would choose this technique again. Conclusions:The sole infusion of remifentanil is a suitable and satisfying single-agent monitored anaesthesia care technique for oocyte retrieval. However, close anaesthetic observation – especially of the respiratory function – is mandatory.

The main purpose of this study was to determine the possible relationship between chromatin condensation (Aniline Blue staining), the morphology of spermatozoa according to strict criteria, and the fertilization, cleavage and pregnancy rate in an intracytoplasmic sperm injection (ICSI) programme. A total of 60 patients were divided into two groups (27 versus 34) according to sperm stainability by Aniline Blue. The first group involved patients having a positive Aniline Blue staining test with 0-29% stained. The fertilization rate in this group was 60.8%, cleavage rate 54.4% and pregnancy rate 18.5%. In the second group in which > 29% spermatozoa were positively stained, the fertilization rate was 62.1%, cleavage rate 62.0% and pregnancy rate 35.3%. There was no statistically significant difference between the two groups. Furthermore, the influence of morphology according to strict criteria after Papanicolaou staining on successful fertilization, cleavage and pregnancy was studied in 85 patients who were divided into two groups according to the percentage of morphologically normal sperm. The fertilization, cleavage and pregnancy rates were 44.21, 63.37, and 39.47% respectively in the first group (< 4%), the corresponding values for the second group (> 4%) were 56.50, 46.04 and 21.21%. There was no significant correlation between the fertilization (P = 0.722), cleavage (P = 0.519) and pregnancy (P = 0.096) rates in either group. This study demonstrates that neither chromatin condensation (Aniline Blue staining) nor morphology could assess the fertilization potential, cleavage and pregnancy rate in an ICSI programme.