Changes in antiproteolytic activity in the blood and cervical mucus after superovulation stimulation]

The function of proteins, peptides, proteases and inhibitors of proteases in modulations of regulation mechanisms of gonadotrophins during the development of ovarian folicles has not been fully explained up to now. We can see difference reactions of ewes to superovulation stimulations in oestrous and anoestrous periods as shown by the variation of the antiproteolytic activity of blood plasma and cervical mucus. Trypsin is used as a model for serine protease, and trypsin inhibitory activity (TIA) was measured from the reduced rate of trypsin hydrolysis of the chromogenic substrate N-alpha-tosyl-L-arginine-4-nitroanilide (TAPA, Bartík et al., 1974). Full hydrolytic activity was determined as a change in absorbency at 405 nm = 1.0 after ten-minute incubation at 25 degrees C and pH = 8.1, and inhibition was expressed in percentage of full activity. Statistical analyses were performed by Student's t-test. Twenty-three ewes in anoestrus and twenty-eight ewes in oestrus were included in this experiment. They were of the Slovak Merino breed, two to three years old, with the mean live weight of thirty to forty kg. The ewes were treated with Ageline vaginal sponges (20 mg chlorsuperlutin/sponge) to provide for synchronization of ovarian activity, in the interval of eleven to twelve days. After removal of sponges, the ewes were stimulated for superovulation as shown in Tab. I. Heparinized blood plasma samples were stored at -25 degrees C. Low molecular TIA activities were determined in HClO4--treated blood plasma. Samples of cervical mucus were taken on cotton-wool tampons which were evaluated in 0.2 M Tris-HCl buffer, pH = 8.1. Figs. 1-6 shows TIA changes in blood plasma and cervical mucus on particular days. Fig. 7 shows the mean values of controls (I-initial) after synchronization (A) and after stimulation (S). TIA changes were different in anoestrous and oestrous periods. Differences in TIA changes in blood plasma and cervical mucus in oestrous period suggested certain local regulation mechanisms of synthesis and/or secretion of this activity in the cervix. Some values of TIA were maximum at the time of expected heat or ovulation and may be some of the factor(s) which influenced fertility of females after superovulation stimulations.     

Changes in the trypsin-inhibiting activity of blood and cervical mucus in sheep after the administration of PGF2 alpha and PMSG in anestrus

The changes in the concentrations of proteolytic enzyme inhibitors after the administration of hormones influencing the activity of the ovaries can cause an imbalance in the development of follicles. Important roles are attributed to serine proteases during the development of follicle, ovulation, and activity of the corpus luteum. In order to find out the changes in the trypsin inhibiting activities (TIA) of the blood plasma and cervical mucus after the administration of PGF2 alpha (100 + 100 micrograms in January and 250 + 250 micrograms in May on the 0th and 11th day) and after the single administration of 750 I. U. PMSG on the 10th day, a 14-day trial was performed to investigate the circadian changes. TIA was determined by means of slowing down the hydrolysis of the synthetic substrate N-alpha-tosyl-p-nitro-anilide (TAPA) by bovine trypsin. When the blood plasma TIA is expressed as the percent of the control values, a large drop is observed on the 11th and 12th days of the January trial (total as well as low-molecular TIA). In the May experiment this drop was recorded only on the 11th day in total TIA (c TIA); low-molecular TIA (n TIA), on the other hand, increased, particularly after the administration of PMSG. The average values of plasma TIA's, recorded in the course of the 14-day trial, increased after the administration of PGF2 alpha (c TIA increased to 109.8 +/- 22.1%, n TIA to 135.42 +/- 54.0% in January; c TIA to 112.59 +/- 50.5%, n TIA to 109.48 +/- 55.9% in May). After the administration of PMSG the TIA values remained higher than the controls (c TIA 107.13 +/- 24.0%, n TIA 133.40 +/- 52.1% in January; c TIA 113.87 +/- 51.3%, n TIA 118.33 +/- 55.6% in May). The average TIA values of the cervical mucus increased after PGF2 alpha administration to 3.06 +/- 1.7% in comparison with the control value of 2.59 +/- 1.1%, and also after the administration of PMSG to 2.82 +/- 1.9% (January, TIA per 1 mg of mucus).     

The effect of PMSG on the trypsin inhibitory activity of the blood and cervical mucus and on the reproductive organs in sheep during anestrus

The effect of the doses of 750, 1000, and 1500 I. U. PMSG (Antex Leo) on changes in the trypsin-inhibition activities (TIA) of the blood plasma and cervical mucus of ewes was found to vary: a) in the times (from PMSG administration) of recording the minimum values of total TIA (72, 48, 24 hours); b) in the values of the average total TIA on individual days of the trial (40.3-141.4% of the control values); c) in the fractions of low-molecular TIA after administration of 750 and 1500 I. U. PMSG, the fractions exhibited minimum values after 24 hours, and of 1000 I. U. after 72 hours (53.0 and 60.4, and 69.7% of the control values, respectively); d) in the highest values of the fractions of low-molecular TIA recorded after 96 hours (96.1-183.7% of the control values); e) in the average values for 96 hours of the trial, which ranged from 64.1% (1500 I. U.) to 77.1% (750 I. U.) for total TIA and from 89.6% (750 and 1000 I. U.) to 122.8% (1500 I. U.) for low-molecular TIA; f) in the ratio of atretic and non-atretic (A/N) tertiary follicles, which grew with increasing PMSG doses; this was indirectly correlated with the average values for total TIA; g) in the weights of uteri and ovaria which increased from 176.8 to 322.3% of the control values after the stimulation; h) in the epithelium thickness of cervix and cervical glands increasing from 118% to 316% of the control values; i) in the average TIA values of cervical mucus ranging from 53.3 to 125% of the control values.     

The effect of prolonged whole-body gamma irradiation of 6,7 Gy and 4,8 Gy (700 and 500 R) on the trypsin inhibiting activity (TIA) of blood, the cervical mucus and the morphological structure of the cervix in ewes

The pattern of changes in the trypsin-inhibition activities (TIA) of blood plasma, cervical mucus and in the morphological structure of cervix was studied in ewes exposed to 60Co radiation for seven and five days, the radiation doses being 6.7 Gy and 4.8 Gy (700 and 500 R). During exposure, the group of ewes irradiated with 4.8 Gy was given the Roboran vitamin premix in addition to standard feed. After termination of irradiation the animals in this trial were given the ampicillin antibiotic (5250 mg). TIA was determined from the retardation of the hydrolysis of the synthetic substrate N-alpha-tosyl-p-nitroanilide (TAPA) by bovine trypsin; the TIA was expressed as the percentage of inhibited trypsin. As found on the day of the termination of irradiation, almost all the studied TIA values of blood plasma and cervical mucus were increased in the irradiated animals, the range being from 103.1 to 155.0% of the levels for non-irradiated ewes. A reduction was recorded only in total TIA of blood plasma in the group irradiated with the dose of 6.7 Gy (83.1% of the values for non-irradiated animals). In the group of animals irradiated with 4.8 Gy and fed no Roboran, the TIA of cervical mucus was observed to decrease to 92.4%. When evaluating the average TIA of different test groups throughout the experiment it was found that all the studied TIA values of the irradiated animals increased within a range from 104.1 to 184.0% of the values for untreated ewes. It was only in the average values of total TIA in blood plasma that, after irradiation with 6.7 Gy, a decrease to 92.7% of the level without irradiation was recorded. It was found during the study of changes in the proportions of glands in the stroma and changes in epithelium thickness in the mucous membrane of cervix uteri that the irradiated ewes had the epithelium thickness reduced to 95.3%-65.5% and that their stromal gland number decreased to 75.4%-79.7% of that recorded in non-irradiated animals. It was only in the group given a Roboran supplement that an increase to 123.7% of the gland number for untreated ewes was recorded on the tenth day after the termination of irradiation.     

The anti-proteolytic activity of blood and ovarian follicular fluid in sheep after stimulation with serum gonadotropin (PMSG) and administration of Antisergon]

The synthesis and secretion of trypsin (trypsin model serine protease) inhibitors are regulated in ovarian follicles by gonadotropins. The superovulation stimulations with 400 IU FSH, 1000 IU PMSG, 1000 IU HCG, 750 IIU PMSG + 750 IU HCG influence in a different way the trypsin inhibiting activities (TIA) of blood plasma (BP) (Figs 1 and 2) and follicular fluid (fig. 3); this points to a possibility of local effects. An increase in the average values of TIA in BP was statistically significant during the whole experiment: P less than 0.05 to P less than 0.001 (following the administration of PMSG+HCG, or PMSG, and HCG); Antisergon administered in 68 hours after PMSG reduced this increase. The changes in the fraction of low-molecular TIA in BP (after BP treatment with perchloric acid) were of converse nature; a decrease in the average values ranged from P less than 0.02 to P less than 0.001 (following PMSG or other stimulations). Antisergon did not influence this decrease. The changes observed on particular days of the trial (Figs. 1 and 2) also indicate different effects of the preparations, mainly of the component LH, which resulted in the occurrence of large nonovulating follicles (greater than 10 mm--"cystic" ones). No such follicles were observed in nonstimulated ewes and after FSH stimulation. The administration of antisergon (goat's antiserum against PMSG) 68 hours after PMSG administration did not prevent their creation. The TIA of follicular fluid (FF) of antral follicles was on average tenfold in comparison with that of blood plasma; and the TIA FF of follicles greater than 10 mm was higher (up to P less than 0.001) than the TIA FF of follicles less than 10 mm. The administration of Antisergon in shorter intervals following PMSG administration (12, 24, 48 and 58 hours) influenced the average values of TIA BP in 120 hours (since PMSG administration) in dependence on time (Tab. I). The effects of Antisergon administered in 12 and 24 hours after PMSG administration on the TIA BP were insignificant if it was administered in 48 and 58 hours the TIA BP increased (P less than 0.02; P less than 0.001) in comparison with the interval of 12 hours. The TIA FF of follicles less than 5 mm, 5-10 mm and greater than 10 mm varied in dependence on the time intervals of Antisergon administration (Fig. 4). The statistical significance of these changes in shown in Tab. II.(ABSTRACT TRUNCATED AT 400 WORDS)     

Changes in antiproteolytic activity in blood and ovarian follicular fluid in 3 breeds of sheep after stimulation of superovulation]

Serine proteases help to regulate the ovarian cycle at different levels and they are subjected to the control of gonadotropic hormones and protease inhibitors. Superovulation stimulations influence the activities of trypsin inhibitors (model serine protease) in blood plasma (BP) and in follicular fluid (FF), and also in dependence on the breed. Trypsin inhibiting activities were determined from the reduced rate of trypsin hydrolysis of a chromogenic substrate (TAPA) and they were determined in percent. A change in absorbancy at 405 nm = 1.0 after 10-minute incubation at 25 degrees C and pH = 8.05 was taken as 100%. The incubation mixture as a sample contained 100 microliters blood plasma or 10 microliters follicular fluid, diluted with gammaglobulin at 1:10. The differences in the trypsin inhibiting activities (TIA) of BP in ewes of the Merino, Tsigai and Wallachian breeds were insignificant, but Agelin synchronization (20 mg chlorsuperlutin per vaginal swab) induced statistically significant differences. The lowest TIA BP was recorded in the Tsigai breed (T), P less than 0.001 in comparison with the Wallachian (W) and Merino (M) breeds. Following the administration of 1,500 IU PMSG, the TIA BP within 120 hours decreased in W (P less than 0.001), it increased in T (P less than 0.1) and in M the changes in the TIA BP were insignificant. The average numbers of ovulations increased from 2.25 +/- 2.5 to 3.0 +/- 1.2 in W; from 0.25 +/- 0.43 to 2.5 +/- 1.6 in T and from 0.00 +/- 0.0 to 2.5 +/- 2.3 in M. Following the single administration of 2,000 IU PMSG after Agelin synchronization, the changes in M ewes were also insignificant, and there were no different responses in pregnant (1st to 2nd month) and nonpregnant ewes. In pregnant T ewes the TIA BP increased after Agelin synchronization and stimulation (P less than 0.01), in nonpregnant ewes these changes were not significant. In W lambing ewes the TIA BP increased (P less than 0.001), the effects of Agelin were greater than those of PMSG. The TIA of follicular fluid (FF) of antral follicles were on average tenfold if compared with BP. After hormonal treatment of ovaries, the TIA FF mostly increased at different levels of statistical significance. The TIA FF of follicles less than 10 mm were lower than in follicles greater than 10 mm (P less than 0.001 for M and W, P less than 0.1 for T).(ABSTRACT TRUNCATED AT 400 WORDS)     

Monoamine oxidase activity in some brain areas of sheep after hormonal stimulation

The influence of hormonal superovulatory preparations Folistiman (450 IU FSH, Spofa, Prague) and serum gonadotropin (1500 IU PMSG, Spofa, Prague) on monoamine oxidase (MAO), the degradative enzyme of catecholamines, was investigated in some areas of the brain regulating reproductive functions (area preoptica of the hypothalamus, pituitary gland, and pineal gland) in ewes with synchronized oestrus (20 mg chlorsuperlutin) during the oestrous period using a radiochemical method. After intramuscular administration of 1500 IU PMSG, marked increase of MAO activity was found in the area preoptica (p < 0.05) and in the pituitary gland (p < 0.01) in comparison with the control group. No change occurred in MAO activity after ovarian stimulation with FSH. Administration of the above superovulatory preparations failed to induce MAO activity in the pineal gland of sheep.     

Morphologic changes in the ovary and histologic changes in the tertiary ovarian follicles in ewes after prolonged irradiation during anestrus

The changes in volume, weight and the histomorphological changes of the tertiary follicles of ewes were studied after protracted irradiation with 4.8 Gy in the anoestrous period by the morphometric and qualitative histological methods. The trial was performed in May with 21 ewes of the Slovak Merino breed, divided into three groups. The first group (five ewes) was control. The second and third groups (each containing eight ewes) were exposed to gamma-rays for five days, the total dose being 4.8 Gy. Within ten days after the treatment, all the irradiated and control ewes were given Ampicillin Spofa per os at a dose of 250 mg per head/day and Roboran Spofa at a dose of 10 g per head/day. The animals were killed by bleeding on the fifth day of irradiation and on the tenth day after the end of the treatment. After killing, the volume and weight of the ovaries were determined and a common histological method was used to cut these ovaries into 7 microns slices in series 70 microns apart. The slices were stained with haematoxylin-eosine and were evaluated by means of light microscopy. After irradiation the weight of the ovaries was found to decrease significantly; however, ovary volume remained unchanged. The atretic and non-atretic tertiary follicles were subjected to qualitative histological differentiation after Marion et al. (1968) and the number of non-atretic follicles was found to have decreased significantly in the irradiated ewes. The late type of atresia contributes most significantly to an increase in the proportion of atretic tertiary follicles. The administration of vitamins after irradiation reduced the occurrence of atretic changes.     

The effect of hormonal stimulation on the follicle population in the ovaries of ewes during estrus and anestrus

In the oestral and anoestral periods, 129 ewes of the Slovak Merino breed were treated with different doses of the stimulants PMSG, PGF2 alpha, HCG, SG and FSH after synchronization by means of Ageline tampons. In the anoestral period, PMSG doses of 750 and 1000 i. u. were found to increase the number of tertiary follicles from 12.3 to 17.2 and 18.6, whereas the dose of 1500 i. u. was found to reduce this number from 18.6 to 13.5. PGF2 alpha increases the average number of tertiary follicles from 12.3 to 22.2 and when it is combined with 750 i. u. PMSG, the increase is to 29.3 tertiary follicles. At higher PMSG doses the ratio of atretic (A) to non-atretic (N) tertiary follicles rose and the number of ovulations increased from 2.5 to 4.0. After synchronization with Ageline and administration of PMSG at the doses of 750 and 1000 i. u., the number of tertiary follicles increased from 16.0 to 18.4 and the number of ovulations from 3.1 to 5.7; the number of atretic tertiary follicles showed no change. In the oestral period, the dose of 1000 i. u. PMSG administered after PGF2 alpha reduced the number of tertiary follicles from 18.9 to 14.5 and increased the number of ovulation from 1.6 to 2.6; an increase was also recorded in the number of atretic tertiary follicles.(ABSTRACT TRUNCATED AT 250 WORDS)     

Enzyme activity in bovine cervical mucus during spontaneous and induced estrus

The purpose of the present research was to compare the enzyme activity of alkaline phosphatase (ALP), lactate dehydrogenase (LDH), α-amylase, α-manosidase, β-N-acetyloglucosaminidase, β-glucuronidase, and β-galactosidase in the cervical mucus of cows during spontaneous and induced estrus. Friesian cows (n = 106) were assigned to 4 groups: 1) no treatment; 2) progesterone releasing intervaginal device (PRID) for 12 days plus pregnant mare serum gonadotrophin (PMSG) at the removal of the PRID; 3) PGF2α2 doses 11 days apart; and 4) PRID for 7 days plus PGF2α 1 dose, 24 hours before removal of the PRID. Fourteen cows were excluded from the trial because of an inadequate quantity of cervical mucus collected or a lost PRID. The cows from the 3 induced estrus groups were artificially inseminated (AI) twice, while those with spontaneous estrus received only a single AI. Cervical mucus samples were collected from all cows 5 to 30 min before the first AI. The results are summarized as follows: 1) ALP and α-amylase activity for spontaneous estrus were similar to those for induced estrus; 2) LDH activity levels during spontaneous estrus were significantly lower (P < 0.001) than that in the P4 and P4+PGF2α induced estrus groups; and 3) glycosidases' activity was significantly lower (P < 0.001) in the spontaneous estrus group than that in the induced estrous groups. In conclusion, the activity of most enzymes in the cervical mucus of cows, in the present study, was significantly different between the spontaneous and the induced estrus groups.     

Qualitative and quantitative changes in the hypothalamus and the ependyma of the 3d cerebral ventricle in sheep after irradiation and hormonal stimulation

In the submitted work we concentrated our attention on the study of the changes in neurosecretion. PAS positive mucopolysaccharides in the hypothalamus and ependyma of the IIIrd cerebral ventricle and on studying the changes in the volume of the cell nuclei of the neurons of nucl. supraopticus (NSO), nucl. hypothalamicus centromedialis (NVM) and nucl. tuberomamillaris (NTM) of sheep after irradiation and after hormonal stimulation. We made our observations on 28 sheep of the Slovak Merino breed, of average live weight 35 to 40 kg, two- and three-years old in the period of physiological anoestrum. The first group of four ewes and the second group of four yearling lambs were controls. The third group of six ewes and the fourth group of six yearlings were exposed to irradiation all over with a dosage of 2.46 Gy (250 R) for a period of five days. To synchronize the oestrum of all the sheep we used agelline sponges which we fitted five days before irradiation. After the irradiation and removal of the sponges we gave the test sheep hormonal stimulation with 3 X 500 i. u. of serum gonadotropin. The fifth group of four ewes and the sixth group of four yearlings were only stimulated without irradiation. The fifth day after stimulation we slaughtered the sheep. We processed the brain samples by the usual histological methods. We carried out karyometric analysis with 3000X magnification and the measurement of 200 cells from one sample. We processed the values obtained mathematically according to Fischer and Inke (1956). We evaluated the quantity of neurosecretion material with a light microscope according to Nakahara (1962). We found that the multiplication of neurosecretion and the increased activity of the PAS reaction in the hypothalamic nuclei studied show that the irradiation and hormonal treatment stimulate the function of the hypothalamic structures. The results of the karyometric analysis in the yearlings also confirm this opinion. In the ewes the inhibitive influence of irradiation was probably manifest. The giving of hormones in combination with irradiation causes the multiplication of ependyma cells and the desquamation of surface layers.     

Dynamic changes in the trypsin-inhibiting activity in cervical mucus, blood and in the histomorphological structure of the uterus in x-irradiated sheep

The effect of ionizing radiation on the reproduction organs of ewes was studied after single local irradiation of ovaries with different X-ray doses. Twenty two ewes of the Slovak Merino breed at the age of two years and at the average live weight of 30 kg were divided into four test groups to be irradiated with the following doses: Group I (n = 6) 4.78 Gy (500 R), group II (n = 6) 9.67 Gy (1000 R), group III (n = 6) 19.14 Gy (2000 R). The ewes of the fourth group were used as the control; they were only laparotomized, otherwise left intact. The trypsin-inhibiting activity of blood plasma and cervical mucus was determined from the retardation of the hydrolysis of low-molecular substrate (TAPA); the thickness of the epithelium of uterine body and cervix was measured microscopically after fixing and staining. The dynamics of changes in the TIA of cervical mucus, in percentual expression, showed differences when measured in the course of 10, 30 and 100 days (1.84 K, aver. for 1st-10th day: 1 = 2.1; 2 = 1.7; 3 = 1.7; 4 = 2.5. Aver. for 10th-30th days: 1 = 1.8; 2 = 1.7; 3 = 1.8; 4 = 1.0. Aver. for 30th-100th day: 1 = 1.7; 2 = 1.1; 3 = 1.2; 4 = 1.5). The dynamics of changes in the thickness of the epithelium of endometrium, uterine horns and uterine cervix had a different nature. A hundred days from irradiation the values showed little difference.(ABSTRACT TRUNCATED AT 250 WORDS)     

The effect of vitamin A on trypsin inhibition activity of the blood and cervical mucus after the effects of hormones on the ovaries of sheep in the estrus period

Vitamin A (50,000 I. U.), administered after oestrus synchronization by PGF2 alpha (2 X 125 micrograms; 1st and 11th day) together with PMSG (750 and 1000 I. U.), had a stronger influence on the changes in the trypsin-inhibition activities (TIA) of the blood plasma, as compared with the effect of PMSG. The changes in the average TIA values within 120 hours after the administration of the stimulating dose were also observed more frequently to depend on vitamin A. After administration of 1000 I. U. PMSG, an increase was recorded only in the values of the fraction of low-molecular TIA, whereas the administration of the combinations of PMSG + vitamin A resulted in an increase of all the TIA's under study. This increase was directly correlated with an increased number of non-atretic tertiary follicles, with an increased number of ovulations (at the same dose of PMSG), and with a reduced ratio of changes in the concentration of progesterone (P) and 17-beta oestradiol (E): P/E = 1.1 after the administration of 1000 I. U. PMSG, P/E = 0.81 after I. U. PMSG + vitamin A, and P/E = 0.90 after 1000 I. U. PMSG + vitamin A. The increase in the average TIA of the cervical mucus is due to the increased secretory activity of the cervical glands rather than to the multiplication of these glands after ovary stimulation.     

Body status and blood metabolites profiles during resumption of postpartum ovarian activity in yak (Poephagus grunniens)

We examined the changes in body weight (BW), back-fat thickness (BFT) and blood metabolites in relation to postpartum (PP) ovarian activity status in twenty female yaks raised under semi-intensive system. BFT and ovarian activities, like follicle development, ovulation (OV) and corpus luteum (CL) development, were monitored from 4 to 15 weeks (wk) PP using ultrasonography. Resumption of ovarian activity was confirmed with ovulation of dominant follicle (DF) and subsequent CL development, and >1 ng/ml progesterone concentration in blood plasma sample after 1week of ovulation. Yaks were further classified as cyclic (with CL), acyclic (without CL), and cystic (with >25 mm follicular cyst; FC). Within 20 weeks PP, 60% yaks resumed cyclic ovarian activity, while 25% failed to initiate cycling activity, and 15% developed follicular cysts. In all categories of yak, BW gradually decreased (p < .05) till nadir; however, nadir reached earlier (p < .05) in acyclic yaks. BFT differed (p < .05) among the yak groups, but it tended to be higher in cyclic yaks as compared to acyclic and cystic. No difference (p > .05) in non-esterified fatty acids (NEFA) values was found among the different categories of yaks, whereas, beta-hydroxy butyrate (BHB) levels were higher in cystic animals as compared to acyclic and cyclic. Blood glucose levels decreased in all yaks during initial 2 weeks after calving. Our findings suggest that yaks with low BW, BFT and glucose levels, and higher BHB values were at risk of delayed resumption of ovarian activity and concomitant development of follicular cysts.     

Changes in the hypothalamus and adjacent ependyma after hormonal stimulation of the ovaries in ewes]

Superovulation treatment leaves alternations in the controlling regions of the hypothalamus and in the adjacent ependyme after ovulation. The test ewes were synchronized with Agelin (20 mg chlorsuperlutin in one vaginal sponge) and stimulated (after the removal of the sponges) with 750 IU PMSG + 750 IU HCG and with 1000 IU HCG and 750 IU PMSG + 5 ml Antisergon (goat antiserum against PMSG), administered 68 hours after PMSG (i.e. 40 hours after HCG). The control ewes were in different stages of the ovarial cycle. The experimental ewes were killed 120 to 130 hours after the start of stimulation. Routine histological techniques were used to treat the brain samples; this treatment was followed by assessment under light microscope. The ependyme epithelium of the third cerebral chamber was studied under scanning microscope. Preparations with different FSH:LH ratios had different effects on the nucleus ventromedialis. Antisergon administration influenced the secretion of NPV (prevented persistent stimulation), which was observed after administration of PMSG + HCG. On the surface of the lower part of the third cerebral chamber the administration of Antisergon slowed the formation of the miniblebs. Supraependyme cells disappeared after stimulation for superovulation.     

Patho-morphologic and histoenzymologic findings after VX (organophosphate) poisoning in sheep]

Sixty-one sheep were administered experimentally a VX chemical (organophosphate) at different rates. In the process of dissection, samples were taken for a histopathological examination. These were samples of brain, liver, kidneys, rumen wall, small intestine, muscles, myocardium, lungs and spleen. For another histochemical examination, samples of tongue, m. longissimus dorsi, jejunum, rumen, liver, kidneys and m. interconstalis were also taken. The activities of alkaline and acid phosphatase, nonspecific esterase, acetylcholinesterase and dehydrogenase of lactic acid were investigated. The most significant changes were found out in the lungs - in form of oedemas and acute catarrhal bronchopneumonias in those animals which died within three days after chemical administration. Sporadic haemorrhages or haemorrhages in form of vast spots were found out under the epicardium. Their range did not relate to the amount of the chemical administered. Rather dilated vessels were observed in the brain and also in the meninges. The histochemical examination showed different activities of enzymes in particular organs of sheep.