Glucose/lactate metabolism of cryopreserved intact bovine ovaries as a novel quantitative marker to assess tissue cryodamage.

For some patients, the autotransplantation of a cryopreserved-thawed intact ovary might be the best option to preserve their reproductive potential after fertility-threatening treatment. The best procedure to successfully cryopreserve a human ovary without inflicting a devastating level of cryodamage is to date unknown. To optimize this procedure, this study developed an assay to monitor the extent of cryodamage inflicted on bovine ovarian tissue by different cryopreservation protocols. The assay measures glucose and lactate metabolism of ovarian tissue fragments in vitro and determines the extent of cryodamage in cryopreserved ovaries. This study tested the cryoprotective effect of two different routes of administration of the cryoprotectant dimethylsulphoxide (DMSO). The cryoprotective effect was assessed in different tissue layers of the ovary, namely the cortex, the subcortex and the medulla. Submersion of intact ovaries in DMSO prior to freezing-thawing resulted in the complete protection of the glucose/lactate metabolism of the cortex, but not of the inner ovarian mass. Perfusion without simultaneous submersion, resulted in partial protection of cortex, subcortex and medulla, while the combination of submersion and perfusion conveyed the highest level of protection for all three ovarian tissue layers.

In vitro 17ß-oestradiol release as a marker for follicular survival in cryopreserved intact bovine ovaries.

Transplantation of cryopreserved intact ovaries from cancer patients is a technically challenging option for restoring fertility after sterilizing cancer therapy. In this paper we describe an assay based on 17ß-oestradiol (oestradiol) production, to monitor the functional damage sustained by the ovarian tissue during the freeze/thawing procedure. To this end, fresh bovine ovarian cortical biopsies were cultured in vitro for 7 days. As a control, the oestradiol release of biopsies that had sustained maximal cryodamage was analyzed. In addition the oestradiol release by cortical biopsies from two ME2SO perfused and cryopreserved intact ovaries was analyzed. Oestradiol production could be measured in culture supernatants, while oestradiol release of maximal cryo-damaged biopsies was at background levels. In vitro oestradiol release by cortical biopsies can be used as a functional marker for cryo-damage and indicates that our assay is suitable to optimize the cryopreservation procedure of intact ovaries.

Optimal perfusion of an intact ovary as a prerequisite for successful ovarian cryopreservation.

BACKGROUND: Cryopreservation and subsequent reimplantation of intact ovaries from cancer patients, offers potentially the best prognosis for restoring fertility after sterilizing cancer treatment. We used bovine ovaries as a model system to explore the perfusion procedure that is required for cryopreservation of intact ovaries. METHODS: The arteria ovarica was cannuled, and ovaries were flushed with Indian ink for 5 min. RESULTS: Successful perfusion of blood vessels was immediately visible macroscopically by a grey to black discoloration of the ovary and was confirmed microscopically, by examining tissue sections. There was no correlation between the time interval from removal of the ovary to the start of the perfusion, and success of perfusion. We determined the percentage of Indian ink-perfused vessels and scored blood vessels in four different size classes. The percentage of perfused vessels increased with an increase in vessel size. In a limited set of preliminary experiments with human ovaries, comparable results were obtained. CONCLUSIONS: Our results show that bovine ovaries are a suitable and adequate model system for optimizing the cryopreservation of human ovaries. As bovine are at least of comparable size to human ovaries, we expect that our results can be extrapolated to the human situation.

The validity of the postcoital test for estimating the probability of conceiving.

OBJECTIVE: We determined the prognostic value of the postcoital test in subfertile couples. STUDY DESIGN: A retrospective study relates the result of the postcoital test to the occurrence of a spontaneous pregnancy. In a fertility center of a university hospital, data from the complete fertility workup of 956 couples who were examined because of infertility due to cervical hostility, male subfertility, or unexplained causes were collected. Statistical analysis was done with Cox regression. RESULTS: The postcoital test result 9 to 12 hours after intercourse is strongly associated with future pregnancy during the first year after intake; the association is also present when the relation with other prognostic variables is taken into account. CONCLUSIONS: The postcoital test result is a strong predictor of possible fertility within the following year.

The prediction of the chance to conceive in subfertile couples.

OBJECTIVE: To develop a model that can predict the chance to conceive spontaneously in subfertile couples. DESIGN: In a cohort study, a consecutive series of patients consulting infertility was followed up. We related information from previous history, physical examination, postcoital tests (PCT), semen analyses, and sperm penetration meter tests with the occurrence of a spontaneous pregnancy. SETTING: Fertility center in a university hospital. PATIENTS: Nine hundred ninety-six couples consulting for infertility due to cervical hostility, male subfertility, or unexplained infertility. INTERVENTIONS: None. MAIN OUTCOME MEASURE(S): Time between intake and occurrence of the first spontaneous pregnancy. RESULTS: Information from the previous history (duration of infertility, primary or secondary female infertility, age of the woman, fertility problems in male's family), the percentage motile sperm in the first semen analysis, and the result of the first PCT are sufficient to predict the chance to conceive. A pocket chart is presented for easy use of the model. CONCLUSIONS: With a limited amount of diagnostic information, the chance to conceive spontaneously can be predicted.

Influence of salazosulphapyridine and 5-aminosalicylic acid on seminal qualities and male sex hormones.

Seminal abnormalities are a common side effect of salazosulphapyridine (SASP) treatment. We evaluated semen qualities and sex hormone concentrations in 11 patients with inflammatory bowel disease (IBD) during SASP treatment and 4 months after replacing SASP with an oral slow-release preparation of 5-aminosalicylic acid (5-ASA). Significant improvement in sperm count (p less than 0.01), morphology (p less than 0.02), and motility (p less than 0.02) could be observed during 5-ASA therapy, in comparison with SASP treatment. Three pregnancies occurred during the study period. The serum concentrations of gonadotrophins, prolactin, and sex hormone-binding globulin were normal in all patients and not significantly different in the two treatment periods. The mean total testosterone concentration decreased significantly (p less than 0.02) after 5-ASA substitution, together with serum albumin (p less than 0.005), although all values remained within normal limits. The apparent free testosterone concentration was not significantly different in the two treatment periods. It is concluded that a significant improvement in semen quality can be obtained in male patients with IBD after replacing SASP with 5-ASA.

Maturation of bovine oocytes transplanted into bovine follicles of follicular and luteal phase.

Forty-one bovine host follicles with guest oocytes were studied. Fifty of these host follicles from the follicular phase and 26 follicles from the luteal phase of the cycle were cultured in a continuous flux system. No hormones were added to the follicular fluid or the culture medium. After culturing there was no close contact between the oocyte complex and the follicular wall. Of the oocytes cultured in the follicular-phase follicles, 27.2% remained in the germinal vesicle stage and 7.1% reached MII, whereas 9.9% of the oocytes cultured in the luteal-phase follicles remained in germinal vesicle phase and 29.5% reached MII. Of the oocytes cultured in the luteal-phase host follicles, 15.2% were fertilized in vitro; none of the oocytes cultured in follicular-phase follicles were fertilized.

Antidiuretic effect of ritodrine with and without beta-adrenergic blockade.

Dose-related effects of ritodrine and ritodrine combined with metoprolol on urinary excretion rate were studied in anesthetized dogs. Urine production was abruptly reduced after a total dose of 4 micrograms.kg-1 of ritodrine. This effect could not be antagonized by metoprolol, although the ritodrine-induced decrease of mean arterial pressure and renal arterial blood flow was significantly inhibited. The possible role of fluid retention during tocolytic treatment, even with beta-adrenergic blockade, in the etiology of pulmonary edema is discussed with a review on recent literature.

Cardiovascular effects of ritodrine are blocked by metoprolol in the anesthetized dog.

Inhibition of ritodrine-induced cardiac and peripheral vascular effects by the beta 1-adrenergic blocker metoprolol, was studied by electromagnetic flow measurements in anesthetized dogs. As expected, metoprolol inhibited the ritodrine-induced/increased cardiac workload and heart rate. Metoprolol also inhibited ritodrine-induced peripheral vasodilation. This leads to questions about the cardiac beta 1-adrenergic selectivity of metoprolol. Regarding earlier studies showing no interference in labor inhibition, the combination of ritodrine and metoprolol might be useful in treatment of preterm labor, while economizing cardiovascular performance centrally as well as peripherally.

Dose-related effects of fenoterol and ritodrine in 10 peripheral vascular beds of the anesthetized dog.

Dose-response curves of the uterine relaxants fenoterol and ritodrine were established for vascular conductance in 10 peripheral arterial beds in anesthetized dogs. Vascular sensitivity was expressed as (E)D50 of conductance response. Fenoterol induced strong vasodilation in all arterial beds except in the kidneys. The gastrointestinal vessels are significantly more sensitive to fenoterol than are muscle and brain vessels. Ritodrine induced strong vasodilation in all arterial beds except in the kidneys and the liver. The vascular sensitivity spectrum of ritodrine differs from that of fenoterol. On a molecular basis fenoterol was 66 times as potent as ritodrine if arterial pressure reduction was used as criterion. When (E)D50 values of vascular conductances of both drugs were plotted against arterial pressure reduction as reference, the gastrointestinal vessels (left gastric, splenic, gastroduodenal, proper hepatic, and superior mesenteric arteries) appeared to be significantly more sensitive to fenoterol than to ritodrine. In contrast, vascular sensitivity in the myocardium, striated muscles, and brain (left coronary, common carotid, femoral, and vertebral arteries) was not statistically different for the two drugs.

Therapeutic abortion: a prospective study. II.

PIP: The psychosomatic and psychological reactions of 75 women applying for a therapeutic abortion were compared with those of 33 women in the same state of pregnancy but not requesting an abortion. The abortion applicants differed significantly from the control group on 8 of the 10 scales tested. The abortion applicants had more prepregnancy health problems and more psychosomatic symptoms than the control group. They also showed more fears for themselves and the baby, less desire for pregnancy, more irritability and tension, less maternal feeling, and more depression and withdrawal than the control group. Parity was not as important in determining responses to the questionnaire as was the patients' abortion status. Literature on the use of pregnancy research questionnaires is reviewed. The possible transience of psychological compared to psychosomatic symptoms is suggested.^ieng