Radioimmunoassay of dibenzazepines and dibenzcycloheptanodienes in body fluids and tissues.

In an attempt to establish a radioimmunoassay (RIA), imipramine and amitriptyline immunogens were prepared; desmethyl derivatives were converted into hemisuccinates, conjugated with bovine serum albumin and used for rabbit immunization. [3H]Amitriptyline (4.3 TBq/mmol) and [3H]imipramine (2.9 TBq/mmol) were prepared by catalytic dehalogenation or reductive alkylation. Dibenzazepines and dibenzcycloheptanodienes were determined in biological fluids by a direct method without deproteinization (lower detection limit of 0.5 microgram.l-1); using high-yield methods they were extracted from cell membranes. Assay of tricyclic antidepressants in humans showed that these substances disappear from plasma much earlier than from cell membranes. Dissociation of the antidepressants bound to cell membranes is slow and their plasma concentrations are not influenced by standing for 2 h at 4 degrees C. During preparing the membranes for binding studies these substances are not removed, and they may affect the results of the binding studies.

Timing of laparoscopic aspiration of preovulatory oocytes in heifers.

Twenty-one heifers were synchronized with PGF(2) alpha and 22 heifers were stimulated with FSH-P in decreasing doses and synchronized with PGF(2) alpha. The beginning of LH rise was observed to be 47.6+/-14.0 h and 35.9+/-1.3 h (P < 0.05) and the peak of LH rise was observed at 53.8 +/- 13.4 h and 40.14+/-1.4 h (P < 0.05) after the luteolyticum administration in the synchronized and superovulated group respectively. The beginning of LH rise was observed 6.9+/-6.7 h and 4.8+/-2.6 h (P < 0.05) and the LH peak was observed 11.8+/-7.7 h and 8.3+/-3.2 h (P < 0.05) after the frist symptoms of oestrus in the synchronized and superovulated group respectively. Ovulation was not observed in stimulated heifers in the period of 23-25 h after the preovulatory LH rise. Compact cumulus oophorus was seen at 30.5%, expanded at 67.0 and partial at 2.5% during this interval of 23-25h. Within this same interval 26.9%, 51.3% and 21.8% oocytes without perivitelline space, with perivitelline space and with extruded first polar body were aspirated respectively. It may be concluded from the reported results that to recover fully mature oocytes for in vitro fertilization, it will be necessary to monitor the preovulatory period of the donor cow in great detail.

[The adenohypophyseal response and effect of GnRh (Dirigestran inj. Spofa) administration in cows in the postpartum period]. / Responze adenohypofýzy a úcinnost aplikace syntetického GnRH (Dirigestran inj. Spofa) u krav post partum.

Radioimmunological investigation (RIA) of the level of pituitary FSH in the peripheral blood of cows after parturition demonstrated that this level increased successively. The FSH value of 32.93 +/- 3.71 ng per 1 ml, recorded the fourth to sixth day post partum, increased to as much as 57.78 +/- 40.98 ng per 1 ml 40 days after parturition. The LH level changed only slightly over the same period (from 1.12 +/- 0.21 ng per ml to 1.72 +/- 1.15 ng per ml). The concentration of progesterone in the blood of the cows was about 0.40 ng per ml during the first 15 days after parturition, but from the 25th day post partum it trebled, on an average (1.53 +/- 1.19 - 1.59 +/- 1.25 ng per ml). The response of the adenohypophysis of the cows to the administration of 200 micrograms of synthetic gonadoreline (spec. Dirigestran inj. Spofa) increased with increasing length of time from parturition. FSH concentration increased less markedly during the first 28 days p.p. and this increase was not uniform in time (the average increase was 1.5 to three times); in the later period the highest increase 300-500% was recorded, as a rule, 120 minutes after GnRH administration. The situation was similar in the increase in LH concentration in peripheral blood, but after the 26th day post partum the average increase in LH accounted for 500 to 600%. The concentration of progesterone in peripheral blood increased by more than 300%, on an average, from the 25th day after parturition. This testifies to the first post partum ovulation and to the formation of a new yellow body in the majority of cows under study.(ABSTRACT TRUNCATED AT 250 WORDS)

[Comparison of the effects of tercleucine and tryptophan superanalogs of LH-RH on gonadotropin secretion in heifers]. / Porovnání úcinku tercleucinového a tryptofanového superanalogu LH-RH na sekreci gonadotropinu u jalovic.

In six heifers without sexual organs the effects were compared of the superanalogues LH-RH [(D-Tle6) and (D-Trp6)] on the secretion of luteinizing hormone (LH) and follicles of stimulating hormone (FSH). The superanalogues were given i. m. at dosages of 10, 50 and 100 micrograms pro toto. Dosages of 50 and 100 micrograms of both superanalogues had a greater effect on the secretion of LH than the dosage of 100 micrograms. The highest average concentration of LH during the time of application (D-Tle6) was recorded in the dosage of 10 micrograms (7.84 ng.ml-1). The average concentrations of LH after application (D-Trp6) were within the range of 5.46 to 9.29 ng.ml-1. Because of the great variability of the concentrations of LH no significant differences were ascertained due to the influence of dosage. From a comparison of the concentrations of LH with the same dosage it emerged that (D-Trp6) after an application of 50 micrograms significantly increased the concentration of LH from the 60th to the 240th minute and thus had a more protracted effect. With dosages of 10 and 100 micrograms of the superanalogues no statistically significant differences were recorded after two hours from application. With (D-Trp6) the higher stimulative effect on the secretion of LH was statistically confirmed. The highest concentrations of follicles of stimulation hormone (FSH) (204.6 and 228.6 ng.ml-1) were found from the 40th to the 100th minute after the application of dosages of 50 and 100 micrograms (D-Tle6). The protracted effect was greatest with a dosage of 100 micrograms (270 mins). (D-Trp6) at a dosage of 100 micrograms caused the greatest effect on the secretion of FSH (226.8 ng.ml-1 for a period of 180 mins). The lower dosages of analogues scarcely differed in response. The dosage of superanalogues has an influence on the concentration of FSH in peripheral blood and on the duration of the protracted effect.

[The effect of ovarian steroids on the pharmacokinetics of Terguride in the rat uterus]. / Vliv ovariálních steroidu na farmakokinetiku Terguridu v deloze krys.

The incorporation of 3H-17-beta oestradiol (3H-E2) and 3H-Tergurid (3H-Te) in the uteri of sexually immature rats was monitored for 16 hours following the treatment of the animals with Tergurid, a semisynthetic ergot alkaloid [chemically acid N-(D-6-methyl-8-isoergolin-I-yl)--N',N'-diethylurea maleate; VUFB-6638]. The differences in the incorporation of 3H-Te in relation to the presence of ovarial steroids (E2 and progesterone) were investigated for the same period of time in the adult ovariectomized rats. In all the time intervals under study, the average values of 3H-E2 activities in the uteri of juvenile rats treated with Tergurid were lower than in the controls. This decrease was statistically significant (P less than 0.05) eight hours after treatment. The time course of the incorporation of 3H-Te in the uteri of the experimental and control animals was about the same. The activities of incorporated 3H-Te in the uteri of adult rats treated with E2 were higher in all the intervals, the difference being statistically significant after two hours and 16 hours (P less than 0.05) and after four hours (P less than 0.01). Progesterone did not induce any significant differences in the incorporation of 3H-Te. These results suggest that the increased incorporation of Tergurid in the uteri of adult rats is due to a direct influence of E2. It has been demonstrated by trials on juvenile rats that Tergurid does not combine with oestrogenic receptors. As suggested by the decreased weights of the uteri of juvenile rats treated with Tergurid, the uterine cells are unable to react to exogenous administration of E2.(ABSTRACT TRUNCATED AT 250 WORDS)

[Fluctuation in the concentration of sex steroids and aflatoxin B1 in the seminal plasma of boars and its relation to sperm production]. / Kolísání koncentrace pohlavních steroidu a aflatoxinu B1 v semenné plazme kancu a jejich vztah k produkci spermatu.

The concentration of testosterone, 17-beta oestradiol and aflatoxin B1 were studied in the semen plasma of 21 boars of four breeds for the period of twelve months. The following spermiological parameters were investigated: semen volume, sperm concentration, percentage of abnormal spermatozoa, and survival of spermatozoa. The fertilizing capacity of ejaculates was evaluated according to the conception rate of sows and gilts after the first insemination, according to the average number of piglets per litter and average number of live-born piglets per litter. The highest aflatoxin B1 residues in sperm were recorded in March to May and were related with aflatoxin concentration in feed ration. The group of boars with fertility disorders had more aflatoxin in their sperm (up to 100 pmol . l-1), lower sperm concentration, lower survival of spermatozoa, and a larger proportion of abnormal spermatozoa. The year season had a significant influence on the concentration of the hormones. The highest average value of testosterone (10.2 +/- 1.28 nmol) was obtained in autumn and lower values were recorded in winter. The changes in 17-beta estradiol concentration were similar to the changes in testosterone content, with the maximum value in November (0.249 nmol X 1(-1]. The boars with reproduction disorders had a significantly lower concentration of 17-beta oestradiol. Significant correlations were found between the concentration of the hormones, semen volume, and sperm concentration. 17-beta oestradiol also had a significant positive correlation to abnormal spermatozoa and to the activity of aspartate aminotransferase.

[Effect of pregnant mare serum on bovine luteal tissue]. / Ucinnost séra brezích klisen na luteální tkáne skotu.

Trials were conducted with four heifers and twelve dairy cows to study the luteotropic effect of the serum of pregnant mares administered at the rates of 2000 to 3000 units. The effect of serum gonadotropin (PMSG) on ovarian activity was compared with the ovarian activity of control animals and ovarian activity after the administration of hypophyseal hormones. The highest progesterone concentration of PMSG-treated cows ranged from 41.9 to 74.3 ng per ml of milk; in the control group this range was between 32.2 and 43.1 ng per ml and in the group of gonadotropin-treated cows between 31.7 and 44.8 ng per ml. In heifers the progesterone levels increased from 3.56 to 4.58 ng per ml of plasma within 24 hours from the administration of PMSG. After 48 hours from administration the average progesterone concentration increased to 11.02 ng per ml of plasma. The animals which did not respond to PMSG administration by the growth of follicles exhibited an increased secretion of progesterone. The administration of hypophyseal hormones in the luteal stage of sexual cycle did not exert any significant influence on progesterone concentration, but stimulated the growth of follicles.

Effect of bovine growth hormone on development of goat mammary tissue in organ culture.

The effect of bovine growth hormone (bGH) on DNA, protein and casein synthesis in goat mammary explants was studied. Growth hormone was unable to stimulate DNA synthesis or potentiate insulin-stimulated DNA synthesis either in high or low oxygen concentrations. In the presence of insulin and cortisol bGH had no effect on the synthesis of explant cytosol proteins. Two preparations of bGH were tested for their effect on casein synthesis. The preparation NIH-GH-B17 at concentration 50 micrograms ml-1 increased casein synthesis similarly as about 0.5 micrograms ml-1 of prolactin. Our preparation, prepared by the method of Prusík and Braun [1967], at concentration 50 micrograms ml-1 had effect lower than 0.05 micrograms ml-1 of prolactin. The lactogenic activity of NIH-GH-B17 was decreased by treatment with specific antisera to bovine prolactin. When submaximum concentration of prolactin (0.05 microgram ml-1) was added, bGH at concentration as low as 0.5 microgram ml-1 had synergistic effect on prolactin stimulated casein synthesis.

[Levels of luteinizing hormone in sheep of various breeds]. / Hladiny luteinizacního hormonu u ovcí ruzných plemen

Determined were the LH values in the course of the sexual cycle which ranged within limits of 1 ng ml-1, with a peak of 60 to 250 ng ml-1 on the day of oestrus. By leaving the lambs with their mothers, the normal cycle is stopped, conclusive differences in the rest levels of LH were found out in the Finnish sheep breed, in Romanov sheep, and in the mutton Merino breed. After ovariectomy there occurred an increase of LH in sheep of the Tsigai breed to 6 ng ml-1 and in sheep of the mutton Merino breed to 18 to 22 ng ml-1.