Antide-induced suppression of pituitary gonadotropin and ovarian steroid secretion in cynomolgus monkeys: premature luteolysis and prolonged inhibition of folliculogenesis following single treatment

Administration of high-dose Antide to ovariectomized monkeys results in rapid, prolonged, and reversible inhibition of gonadotropin secretion. The present study examined whether similar long-term control would be manifested in the menstrual cycle of intact primates. Antide administration at a dose of either 3.0 or 18.0 mg/kg induced rapid suppression of bioassayable LH concentrations, precipitating a concurrent fall in serum progesterone concentrations from 7.9 +/- 3.6 and 5.8 +/- 1.0 ng/ml (mean +/- SEM) on the day of injection to 0.6 +/- 0.2 and 0.5 +/- 0.1 ng/ml by 2 days post-treatment, respectively. This Antide-induced luteolysis was accompanied by the premature onset of menses within 3 days. The next menses following Antide administration was delayed. Ultimately, folliculogenesis culminating in normal follicular-phase estradiol production, ovulation, and subsequent normal luteal-phase progesterone production did occur in all treated monkeys. Menses resumed 54 +/- 9 and 75 +/- 13 days after treatment with 3.0 and 18.0 mg/kg Antide, respectively. No allergic cutaneous or peripheral reactions were seen, even at the highest dose of Antide. Thus, the long duration of action of high-dose Antide reported earlier in ovariectomized monkeys is also demonstrated in intact primates. These findings, along with the apparent absence of histamine-release effects even at high doses, suggest that Antide is a GnRH antagonist deserving clinical evaluation.     

Subcutaneous implantable infusion device for chronic intravenous pulsatile gonadotropin-releasing hormone treatment

Preliminary data indicate the potential utility of an implantable subcutaneous device that facilitates chronic intravenous infusion of pulsatile gonadotropin-releasing hormone (GnRH) for ovulation induction. GnRH distribution curves were congruent in control monkeys and those with implanted devices. Tissue tolerance was good in this brief trial. These findings suggest that use of this or a similar implantable device be considered for chronic GnRH administration in human pulse therapy.     

Anaerobic degradation of papermill sludge in a two-phase digester containing rumen microorganisms and colonized polyurethane foam

Summary The use of reticulated polyurethane foam as a support material for the immobilization of methanogenic associations and its application to the anaerobic treatment of fine particulate solid wastes was investigated. The colonization of polyurethane support particles in a continuous upflow reactor fed on a mixture of acetate, propionate and butyrate, was both rapid and dense. The combination of rumen microorganisms and colonized support particles in a two-phase digester resulted in an efficient anaerobic decomposition of papermill sludge.     

Transitory increases of luteinizing hormone and follicle-stimulating hormone following luteectomy in hemiovariectomized primates significantly increase progesterone secretion in vitro by the subsequent corpus luteum

Ten chronically hemiovariectomized cynomolgus and rhesus monkeys were luteectomized 5.5 +/- 0.3 days after the midcycle luteinizing hormone (LH) and follicle-stimulating hormone (FSH) surge in two consecutive cycles. The corpus luteum (CL) was removed, weighed, dispersed with collagenase and the luteal cells counted. Luteal cells (50,000/ml) were incubated in Ham's F10 medium for 3 h at 37 degrees C either in the presence or absence of 100 ng/ml human chorionic gonadotropin (hCG). Daily blood samples were taken from the monkeys throughout the study for determination of LH, FSH, estradiol (E2) and progesterone levels. Within 5 days following each luteectomy (LX), all monkeys responded with a significant increase in FSH and LH (P less than 0.05). Ovulatory LH/FSH surges occurred 14.4 +/- 0.5 days after the first LX. Hormonal profiles of serum progesterone prior to the first and second LX, CL weight and number of luteal cells/CL were similar (P greater than 0.05). However, luteal cells obtained at the second LX produced more progesterone (P less than 0.05) in vitro under basal and hCG-stimulated conditions than cells from the first LX. The areas under the LH and FSH curves following the first LX were highly correlated (P less than 0.05) with the in vitro progesterone production following the second LX. Thus, the monkeys with the largest areas under the LH and FSH curves subsequently had the highest in vitro progesterone production.     

Maintenance of ovulatory menstrual cycles in chronically cannulated monkeys: a vest and mobile tether assembly

We report maintenance of apparently normal ovulatory menstrual cycles in cynomolgus monkeys up to 3 months after their placement in a vest and mobile tether assembly, with or without chronic cannulation via a femoral to vena caval catheter. Since menstrual cyclicity and characteristic hormonal profiles of the principal ovarian steroids remained normal in these monkeys, use of the vest and mobile tether apparatus described here permits long-term study of the hypothalamic-pituitary-ovarian axis during chronic cannulation without introducing overt aberrancies as a part of the experimental model.     

Immunocytochemical studies of hamster oocyte activation

By indirect immunofluorescence, using rabbit anti-heparin-binding placental protein (HBPP) antiserum, we studied HBPP expression by physiologically and non-physiologically (microsurgically) activated hamster gametes. Whereas mature gametes (sperm, metaphase II oocytes) were negative, in vivo conceived preimplantation embryos, from pronuclear to two- and four-cell stages, were HBPP positive. No HBPP was demonstrated in the zona pellucida, but HBPP-dependent immunofluorescence was localized in the perivitelline space. Oocytes incubated with hyaluronidase demonstrated variable responses from negative to positive. (Diluent or sperm) microinjected oocytes were all activated and HBPP positive within 4 h after stimulation. Thus neither activation by microinjection nor HBPP expression required paternal gametes. These kinetics suggest that HBPP may be a cortical granule secretogogue which can be applied to monitor oocyte responses during in vitro manipulations.     

Progesterone production by luteal cells isolated from cynomolgus monkeys: effects of gonadotropin and prolactin during acute incubation and cell culture

Corpus luteum function in cynomolgus monkeys (Macaca fascicularis) during the menstrual cycle and immediately following parturition was evaluated through in vitro studies on progesterone production by dispersed luteal cells in the presence and absence of human chorionic gonadotropin (hCG) or human prolactin (hPRL). Luteal cells isolated between days 17-20 of the menstrual cycle secreted progesterone (P) during short-term incubation (21.6 +/- 1.2 ngP/ml/5 X 10(4) cells/3 hr, X +/- S.E., n = 7) and responded to the addition of 1-100 ng hCG with a significant (p less than 0.05) increase in P secretion. Cells removed the day of delivery secreted large, but variable (27.9-222 ng/ml, n = 4) amounts of P during short-term incubation. Moreover, hCG (100 ng/ml) stimulation of P production by cells at delivery (176 +/- 19% of control) was less than that of cells from the cycle of (336 +/- 65%). The presence of hPRL (2.5-5000 ng/ml) failed to influence P secretion by luteal cells during short-term incubation in the presence or absence of hCG. P production by luteal cells obtained following delivery declined markedly during 8 days of culture in Ham's F10 medium: 10% fetal calf serum. Continual exposure to 100 ng/ml of hCG or hPRL failed to influence P secretion through Day 2 of culture. Thereafter hCG progressively enhanced (p less than 0.05) P secretion to 613% of control levels at Day 8 of culture. In contrast, hPRL significantly increased P secretion (163% of control levels, p less than 0.05) between Day 2-4 of culture, but the stimulatory effect diminished thereafter. The data indicate that dispersed luteal cells from the cynomolgus monkey provide a suitable model for in vitro studies on the primate corpus luteum during the menstrual cycle, pregnancy, and the puerperium, including further investigation of the possible roles of gonadotropin and PRL in the regulation of luteal function in primates.     

In. vitro evaluation of corpus luteum function of cycling and pregnant rhesus monkeys: Progesterone production by dispersed luteal cells

Corpus luteum function in the cycling and the pregnant rhesus monkey (Macaca mulatta) was evaluated through short term $\text{in vitro}$ studies of progesterone production by suspensions of collagenase-dispersed luteal cells in the presence and absence of exogenous gonadotropin (human chortonic gonadotropin, HCG). Cells from mid-luteal phase of the menstrual cycle secreted progesterone, as measured by accumulation of this hormone in the incubation medium, and responded to the addition of 100 ng HCG/ml with a marked increase in progesterone secretion above basal level ($\text{63.7 ± 13.1}\text{versus}\text{24.7 ± 5.5}\text{ng progesterone}\text{/}\text{ml}\text{/5 × 10}{}^4\text{cells}\text{/ 3}\text{hr}\text{,}\text{X}\text{± S.E.,}\text{n}\text{= 6;}\text{p}\text{< 0.05}$). However, luteal cells from early pregnancy (23–26 days after fertilization) secreted significantly less progesterone than cells of the non-fertile menstrual cycle (3.6 ± 2.4 versus 24.7 ± 5.5 ng/ml/5 × 10⁴ cells/3 hr, n = 3; p < 0.05) and did not respond to HCG with enhanced secretion. By mid-pregnancy (108–118 days gestation) luteal cells exhibited partially renewed function, and near the time of parturition (163–166 days gestation) basal and HCG-stimulated progesterone secretion (30.2 ± 5.6 and 63.0 ± 13.0 ng/ml/5 × 10⁴ cells/3 hr, respectively; n = 3) was equivalent to that of cells from the luteal phase of the non-fertile menstrual cycle. The data suggest that following a period around the fourth week of gestation, when steroidogenic activity is markedly diminished, the corpus luteum of pregnancy progressively reacquires its functional capacity and at term exhibits gonadotropin-sensitive steroidogenesis similar to that of the corpus luteum of the menstrual cycle.