Nuclear cyclophilins affect spliceosome assembly and function in vitro.

Cyclophilins are ubiquitously expressed proteins that bind to prolines and can catalyse cis/trans isomerization of proline residues. There are 17 annotated members of the cyclophilin family in humans, ubiquitously expressed and localized variously to the cytoplasm, nucleus or mitochondria. Surprisingly, all eight of the nuclear localized cyclophilins are found associated with spliceosomal complexes. However, their particular functions within this context are unknown. We have therefore adapted three established assays for in vitro pre-mRNA splicing to probe the functional roles of nuclear cyclophilins in the context of the human spliceosome. We find that four of the eight spliceosom-associated cyclophilins exert strong effects on splicing in vitro. These effects are dose-dependent and, remarkably, uniquely characteristic of each cyclophilin. Using both qualitative and quantitative means, we show that at least half of the nuclear cyclophilins can act as regulatory factors of spliceosome function in vitro. The present work provides the first quantifiable evidence that nuclear cyclophilins are splicing factors and provides a novel approach for future work into small molecule-based modulation of pre-mRNA splicing.

Skin shop: A new model for high-volume skin cancer care.

INTRODUCTION: Non-melanoma skin cancer incidence is increasing in New Zealand. Increased cost of care has led to service pressure and a review of models of care. A high-volume skin surgery service at Waikato Hospital has been developed to reduce service costs. This study examines the oncological safety of the new model. METHODS: Prospective data for all skin lesions excised were collected from December 2014 to December 2016. Primary outcomes were rate of complete excision, rate of incomplete excision and rate of narrow excision. RESULTS: A total of 2076 lesions were excised: 92% were complete, 4.2% were narrow and 3.2% were incomplete. CONCLUSION: The rate of narrow and incomplete excisions was low in a service delivered by supervised surgical registrars. The Skin Shop model is safe, inexpensive and suitable for adaptation to safely reduce the cost of skin cancer surgery.

Population dynamics models in plant-insect herbivore-pesticide interactions.

We consider several population dynamics models in investigating data from controlled experiments with aphids in broccoli patches surrounded by different margin types (bare or weedy ground) and three levels of insecticide spray (no, light, or heavy spray). We carry out parameter estimation computations along with statistical analysis to compare autonomous versus non-autonomous model dynamics. We conclude with a brief discussion of some not-so-subtle pitfalls that can arise when using quantitative measures of model fit-to-data to make biological inferences as well as offer a positive example of how one might combine a priori biological hypothesis and intuition with rather sophisticated (from a field biology viewpoint) mathematical methodologies to suggest synergisms.

Concentration of GnRH receptor and GnRH receptor mRNA in pituitary tissue of orchidectomized sheep: effect of oestradiol, progesterone, and progesterone withdrawal.

The effect of progesterone (P4) and P4 withdrawal on oestradiol (E2)-induced change in gonadotrope responsiveness (GR) and concentration of GnRH receptor and GnRH receptor mRNA in pituitary tissue of orchidectomized sheep (wethers) was determined. Thirty wethers were assigned at random to one of six treatment groups (n = 5 animals/group). Wethers received E2 (2 micrograms/h; groups 2, 4, and 6) in 10% ethanol-saline (vehicle), or vehicle alone (groups 1, 3, and 5), as a continuous infusion for 24 h beginning 7 days after insertion (s.c.) of blank (groups 1 and 2) or P4-containing (groups 3-6) implants. The effect of P4 withdrawal was assessed by removing P4-containing implants at the beginning of vehicle (group 5) or E2 (group 6) infusion. Gonadotrope responsiveness (increase in serum LH induced by 500 ng GnRH, i.v.) was assessed at the end of infusion. In a companion study, anterior pituitary tissue was collected at the end of the 24-h infusion period. Infusion of E2 increased (P < 0.05) GR relative to GR noted in control wethers receiving vehicle alone. The magnitude of E2-induced augmentation of GR was not affected by concurrent administration of P4 or P4 withdrawal. Pituitary tissue concentrations of GnRH receptor and GnRH receptor mRNA were significantly reduced in wethers implanted with P4. This P4-induced decrease in tissue concentration of GnRH receptor and GnRH receptor mRNA was not reversed during the 24-h period after P4 withdrawal. Steady-state concentrations of GnRH receptor and GnRH receptor mRNA were significantly increased by E2. However, the magnitude of oestrogen-induced increase in tissue concentrations of GnRH receptor mRNA was not significantly affected by P4 or P4 withdrawal. Conversely, concurrent progestin stimulation potentiated the E2-induced augmentation of tissue concentrations of GnRH receptor. However, this P4-induced potentiation of the oestrogenic response was not evident 24 h after removal of the P4-containing implants. Steady-state concentrations of mRNA encoding the LH beta and FSH beta subunits were reduced (P < 0.05) by P4. Infusion of E2 had a similar affect. These data indicate that prolonged progestin stimulation leads to a decrease in tissue concentrations of GnRH receptor and GnRH receptor mRNA. This P4-induced suppression of GnRH receptor activity is not reversed within 24 h of P4 withdrawal. In addition, the increase n steady-state concentrations of GnRH receptor and GnRH receptor mRNA induced by E2 is not compromised by concurrent progestin stimulation.

Development of the ventilatory response to hypoxia in Swiss CD-1 mice.

We examined developmental changes in breathing pattern and the ventilatory response to hypoxia (7.4% O(2)) in unanesthetized Swiss CD-1 mice ranging in age from postnatal day 0 to 42 (P(0)-P(42)) using head-out plethysmography. The breathing pattern of P(0) mice was unstable. Apneas were frequent at P(0) (occupying 29 +/- 6% of total time) but rare by P(3) (5 +/- 2% of total time). Tidal volume increased in proportion to body mass ( approximately 10-13 ml/kg), but increases in respiratory frequency (f) (55 +/- 7, 130 +/- 13, and 207 +/- 20 cycles/min for P(0), P(3), and P(42), respectively) were responsible for developmental increases in minute ventilation (690 +/- 90, 1,530 +/- 250, and 2,170 +/- 430 ml. min(-1). kg(-1) for P(0), P(3), and P(42), respectively). Between P(0) and P(3), increases in f were mediated by reductions in apnea and inspiratory and expiratory times; beyond P(3), increases were due to reductions in expiratory time. Mice of all ages showed a biphasic hypoxic ventilatory response, which differed in two respects from the response typical of most mammals. First, the initial hyperpnea, which was greatest in mature animals, decreased developmentally from a maximum, relative to control, of 2.58 +/- 0.29 in P(0) mice to 1. 32 +/- 0.09 in P(42) mice. Second, whereas ventilation typically falls to or below control in most neonatal mammals, ventilation remained elevated relative to control throughout the hypoxic exposure in P(0) (1.73 +/- 0.31), P(3) (1.64 +/- 0.29), and P(9) (1. 34 +/- 0.17) mice but not in P(19) or P(42) mice.

The Older Adult Health and Mood Questionnaire: a measure of geriatric depressive disorder.

Depressive disorders are a common problem in late life, requiring consistent and reliable screening by clinicians. Such screening should be based upon the most recent criteria available to facilitate diagnostic and nosologic agreement. This study presents a new screening instrument for geriatric depressive disorders based upon DSM-III-R criteria and the known differences in the display of depression that occur among older persons. The Older Adult Health and Mood Questionnaire (OAHMQ) is a 22-item questionnaire with sound psychometric properties and high validity. Its validation included comparisons with diagnostic assessments by geriatrically-trained psychiatrists and psychologists, and factor analysis. Using a three-part division of "normal," "clinically significant depressive symptoms," and "probable major depression," its sensitivity is .80 and its specificity is .87 for major depression, and .92 and .87, respectively, for any depressive disorder. Separate scoring of affective symptoms versus other symptoms of depression (i.e., physiologic, cognitive, or behavioral) can be useful for diagnostic and research purposes.

Concentrations of bioactive and immunoactive luteinizing hormone in bovine anterior pituitary tissue.

Tissue concentrations of LH as determined by radioimmunoassay (RIA) may not accurately measure actual changes that could occur in biological activity of the hormone. To examine this possibility, pituitary homogenates from 135 beef cows in various physiological states were analyzed for content of LH by both RIA and an in vitro bioassay. The ratio of biological/immunological active concentrations of LH remained constant (.52 +/- .02) even though tissue concentrations of immunoactive LH differed among groups. Tissue concentrations of bioactive LH were linearly related to, and highly correlated with (P less than .001), tissue concentrations of immunoactive LH. These data indicate that only a fraction of the immunoactive LH in the bovine pituitary is biologically active. However, this fraction does not vary with the reproductive status or plane of nutrition.

Effect of stress-like concentrations of cortisol on the feedback potency of oestradiol in orchidectomized sheep.

The effect of stress-like concentrations of cortisol on oestradiol-induced change in LH secretion and GnRH receptor expression was evaluated in orchidectomized sheep (wethers). Twenty-four wethers were assigned at random to one of the four treatment groups in a 2x2 factorial design (n=6 wethers/group). Wethers received cortisol (90 microg/kg/h; groups 2 and 4) or a comparable volume of cortisol delivery vehicle (groups 1 and 3) by continuous infusion for 48 h. During the final 24 h of infusion, wethers received oestradiol (6 ng/kg/h; groups 3 and 4) or oestradiol delivery vehicle (groups 1 and 2). The pattern of LH secretion was assessed during a 3-h period of intensive blood collection beginning 21 h after initiation of oestradiol infusion. Although neither cortisol nor oestradiol alone affected (P>0.05) mean serum concentration of LH or LH pulse frequency, serum LH and the frequency of secretory episodes of LH were significantly reduced (P<0.05) in wethers receiving cortisol and oestradiol in combination. Anterior pituitary tissue was collected at the end of the infusion period. Oestradiol increased (P<0.05) tissue concentrations of GnRH receptor and GnRH receptor mRNA. Although cortisol alone did not affect (P>0.05) basal concentrations of receptor or receptor mRNA, the magnitude of oestradiol-induced increase in GnRH receptor and GnRH receptor mRNA was significantly reduced in wethers receiving cortisol and oestradiol concurrently. Conversely, steady-state concentrations of mRNA encoding the LHbeta and FSHbeta subunits were increased (P<0.05) in wethers receiving cortisol. These observations demonstrate that stress-like concentrations of cortisol act in concert with oestradiol to suppress LH secretion. In addition, cortisol blocks oestradiol-dependent increase in pituitary tissue concentrations of GnRH receptor and GnRH receptor mRNA.

Effect of duration of infusion of stress-like concentrations of cortisol on follicular development and the preovulatory surge of LH in sheep.

Stress-like levels of cortisol suppress follicular growth and development and block or delay the preovulatory surge of LH when cortisol is continuously administered during the late luteal and early follicular phases of the ovine oestrous cycle. We postulated that cortisol infusion of shorter duration would have a similar effect. To test this hypothesis the oestrous cycles of mature ewes were synchronized using progestin-treated vaginal pessaries. Ewes were randomly assigned to one of four treatment groups. Animals received cortisol (0.1mg/kg/h; n=8) or vehicle alone (n=8) beginning 5 days before, and continuing for 5 days after, pessary removal (PR). Additional groups received cortisol only during the 5 days period before (n=7), or the 5 days period after (n=8), PR. Continuous delivery of cortisol established stable serum concentrations of cortisol of 72.0+/-2.5ng/ml within 6h of initiation of infusion. Serum concentrations of oestradiol increased progressively during the period after PR in control animals receiving vehicle alone and the preovulatory surge of LH was evident in all control animals (eight of eight) 55.5+/-5.0h after PR. In contrast, follicular development and the preovulatory surge of LH were evident during the period of cortisol infusion in only one of eight animals receiving stress-like levels of cortisol over the entire 10-day infusion period. Similarly, neither follicular development nor surge-like secretion of LH were evident during the infusion period in animals (zero of eight) receiving cortisol during the 5-day period after PR. This cortisol-dependent suppression of ovarian activity in sheep receiving stress-like levels of cortisol during the 5 days after PR was temporary and follicular development, the ovulatory surge of LH, and subsequent luteal function were evident in six of eight ewes after cessation of cortisol delivery. Similarly, follicular development and the preovulatory surge of LH were noted within 5 days after PR in four of seven ewes receiving cortisol only during the 5-day period prior to PR. Collectively, these data indicate that stress-like levels of cortisol reduce fertility of sheep by suppressing follicular development and the preovulatory surge of LH. Additionally, cortisol delivery during the follicular phase has a more profound suppressive effect on follicular development than cortisol administration during the luteal phase.

Reproductive function and feedlot performance of beef heifers actively immunized against GnRH1.

Two trials were conducted to examine reproductive function and feedlot performance by heifers after active immunization against GnRH. In trial 1, heifers were not immunized or were immunized with one of three doses of a GnRH-KLH (keyhole limpet hemocyanin) conjugate in Freund's complete adjuvant. Antibodies against GnRH were not detectable in non-immunized heifers (n = 9). However, antibodies against GnRH were noted in all immunized animals (n = 30) within 8 wk of primary immunization; anti-GnRH antibody concentrations were at a maximum 16 to 20 wk after immunization. This increased anti-GnRH titer was associated with a decreased serum concentration of progesterone. Ovarian and uterine weight and tissue concentrations of LH and GnRH receptor were reduced (P less than .05) by immunoneutralization of GnRH. Similarly, immunization against GnRH reduced (P less than .05) weight gain during feedlot confinement. In trial 2, feedlot performance after insertion of anabolic steroid implants (Synovex H) was evaluated in non-immunized heifers (n = 15), heifers actively immunized against GnRH-KLH (n = 15) or KLH alone (n = 15), or non-immunized heifers treated with melengestrol acetate (MGA; n = 15). Serum concentrations of progesterone were depressed in anti-GnRH and MGA-fed groups, but ovarian and uterine weights were depressed (P less than .05) only in heifers immunized against GnRH. Total weight gain and gain during the final 4 wk of confinement did not differ (P greater than .05) among groups with steroid implants. The GnRH-KLH conjugate is an effective immunogen in heifers, leading to suppression of reproductive activity. The depression of weight gain that attends development of anti-GnRH titers may be reversed by use of implants that contain anabolic steroids.

Feedlot performance of steers and bulls actively immunized against gonadotropin-releasing hormone.

Feedlot performance and testicular and pituitary function were assessed in cattle actively immunized against GnRH. In Trial 1, 50 steers were either unimmunized (n = 10), actively immunized against keyhole limpet hemocyanin (KLH; n = 10), or immunized against a GnRH-KLH conjugate (n = 30). Fifteen of 30 steers immunized against GnRH-KLH received a secondary immunization 8 wk after primary immunization. Antibodies against GnRH were not evident in unimmunized steers or steers actively immunized against KLH. Antibodies against GnRH were noted in all immunized animals (n = 30) within 6 wk of primary immunization and anti-GnRH antibody concentrations became maximal 20 to 24 wk after immunization. The increasing anti-GnRH titer in immunized steers was associated with decreasing serum concentrations of LH. Serum concentrations of LH were depressed (P less than .05) within 8 wk of primary immunization and reached a nadir by wk 20. The patterns of increase in GnRH titer and decrease in serum concentrations of LH did not differ (P greater than .05) in animals receiving primary immunization alone or primary and secondary immunization. Feedlot performance and carcass quality were not affected (P greater than .05) by immunization against KLH or the GnRH-KLH conjugate. In Trial 2, 60 bull calves (mean weight = 325.2 +/- 2.8 kg) were randomly assigned to a 2 x 3 factorial experiment. The two classes (n = 30) were 1) unimplanted and 2) implanted with Synovex-S. The three treatments (n = 20) were 1) intact control, 2) actively immunized against GnRH, and 3) castrate.(ABSTRACT TRUNCATED AT 250 WORDS)

Concentration of gonadotropin-releasing hormone receptor messenger ribonucleic acid in pituitary tissue of orchidectomized sheep: effect of passive immunization against gonadotropin-releasing hormone.

The effect of immunoneutralization of gonadotropin-releasing hormone (GnRH) on LH secretion and concentrations of GnRH receptor, GnRH receptor mRNA, and gonadotropin subunit mRNA in pituitary tissue of orchidectomized sheep (wethers) was assessed. Thirty-six wethers were assigned at random to one of six treatment groups (six wethers per group). Thirty wethers (groups 2 to 6) received 200 ml, (i.v.) of anti-GnRH antisera at passive immunization (PI). Anterior pituitary tissue was collected .5, 1, 2, 4, or 8 d after PI from wethers in groups 2 to 6, respectively. Pituitary tissue was also collected from unimmunized wethers (Group 1). Intravenous administration of anti-GnRH sera increased anti-GnRH activity to 69.1 +/- 7% (percentage of total 125I-labeled GnRH bound by a 1:1,000 serum:GEL-PBS dilution) within 1 h of PI. Anti-GnRH activity declined gradually during the period after PI, and 8 d after PI anti-GnRH activity was 57.2 +/- 1.7%. Serum concentration of LH was significantly reduced, relative to the pretreatment (16.1 +/- 1.8 ng/mL) level, within 4 h (7.6 +/- 1.5 ng/mL) of PI, and the LH level was 10% of the pretreatment concentration 8 d after PI (1.6 +/- 0.2 ng/mL). Steady-state concentration of GnRH receptor mRNA decreased progressively during the period after PI and was significantly reduced, relative to the level in unimmunized control wethers (.44 +/- .03 pg/micron total RNA) d after PI. Tissue concentrations of GnRH receptor and mRNA for the alpha, LH alpha, and FISH beta subunits were also reduced (P < .05) by PI. These data indicate that maintenance of steady-state concentrations of GnRH receptor and GnRH receptor mRNA requires continued GnRH stimulation.

Testes function and feedlot performance of bulls actively immunized against gonadotropin-releasing hormone: effect of age at immunization.

Testes function, feedlot performance, and carcass traits were evaluated in bulls actively immunized against gonadotropin-releasing hormone (GnRH) at different ages. Bull calves were randomly assigned to one of seven treatment groups (n = 15 calves/group). Calves were unimmunized (Group 1), immunized at 1.5, 4, 7, or 12 mo of age with a GnRH-keyhole limpet hemocyanin (KLH) conjugate (Groups 2 to 5, respectively), or castrated at 4 mo of age (Groups 6 and 7). Immunized bulls did not receive a secondary, or booster, immunization. Calves in group 6 received Synovex-C at castration and Synovex-S at weaning and feedlot entry. Anti-GnRH titer was evident at slaughter in all immunized bulls. However, the final immune response of bulls immunized at 1.5 mo was significantly lower than the response of bulls immunized at later stages of development. Final scrotal circumference and testis weight in bulls immunized at 4, 7, or 12 mo of age were significantly reduced relative to unimmunized bulls. The final live weight, feedlot gain, and carcass weight of immunized and unimmunized bulls did not differ (P > .05) from the same parameters in steers implanted with Synovex. Longissimus muscle area, marbling score, and backfat thickness did not differ between immunized and unimmunized bulls. The sex class score of the carcasses of immunized bulls did not differ from the score of steer carcasses. In contrast, a significantly higher proportion of carcasses from unimmunized bulls graded as bullock carcasses. Taken together, these data indicate that a single immunization against GnRH at 4 to 12 mo of age results in significant attenuation of testicular growth in bulls. These data also demonstrate that immunization against GnRH reduces the masculinity of carcasses from bulls, but does not affect feedlot performance, longissimus muscle area, marbling score, or backfat thickness. These results suggest that single immunization with the GnRH-KLH conjugate may have practical utility as a noninvasive alternative to surgical castration in management of beef cattle.

Testis function and feedlot performance of bulls actively immunized against gonadotropin-releasing hormone: effect of implants containing progesterone and estradiol benzoate.

The effect of implants containing anabolic steroid (Synovex) on testis function, feedlot performance, and carcass traits was evaluated in bulls, steers, and bulls actively immunized against gonadotropin-releasing hormone (GnRH). Ninety-one Hereford or Hereford x Red Angus bull calves were randomly assigned to a 3 x 2 factorial experiment. The treatments were: 1) unimmunized control (n = 30), 2) actively immunized against GnRH (n = 31), or 3) castrate (n = 30). The classes were 1) unimplanted (n = 45) or 2) implanted (n = 46) with Synovex. Animals in treatment Groups 2 and 3 were either immunized with a GnRH-keyhole limpet hemocyanin (KLH) conjugate or castrated at 3.6 +/- .1 mo of age. Animals assigned to Class 2 received Synovex-C at the time of initial treatment and Synovex-S at weaning and feedlot entry. Animals in treatment Group 2 received a booster immunization at feedlot entry. Anti-GnRH titer was evident at weaning and remained elevated to slaughter in all (31 of 31) immunized animals. Anti-GnRH titer was not evident in steers or unimmunized bulls. Immunization and (or) Synovex decreased (P < .05) serum concentrations of testosterone (T) and scrotal circumference (SC) at weaning. Similarly, SC, testis weight, and tissue concentrations of spermatozoa at slaughter were significantly reduced by immunization, Synovex, or the immunization/Synovex combination. Immunization, but not Synovex, reduced (P < .05) serum levels of T at slaughter. The masculinity of immunized bulls was less (P < .05) than that of control bulls.(ABSTRACT TRUNCATED AT 250 WORDS)

Dietary restriction reduces the rate of estradiol clearance in sheep (Ovis aries).

Three experiments were designed to test the effect of dietary restriction on clearance of 17beta-estradiol (E(2)) in sheep. A preliminary experiment examined the effect of a 4-d fast on the rate of E(2) clearance in wethers. The second experiment tested the hypothesis that either long-term restriction (7 wk) or a 5-d fast would increase steroid-binding capacity of serum by increasing the concentration of sex hormone-binding globulin (SHBG) in the blood of ovariectomized ewes. In Exp. 3, we hypothesized that nutrition-dependent regulation of E(2) clearance by the liver would result in divergence in biliary extraction of E(2) in fed and fasted wethers receiving comparable levels of exogenous E(2). A marked difference in E(2) clearance between fed and fasted wethers was noted in the preliminary study. Relative to ad libitumfed wethers, a 4-d fast decreased E(2) clearance by 52%. Serum concentrations of SHBG were increased in long-term energy-restricted and fasted ewes, relative to the concentration in maintenancefed ewes (P = 0.015). Furthermore, a 5-d fast nearly doubled serum steroid-binding capacity in wethers. The E(2) concentration in bile was 2 times greater in fasted than in fed wethers. This fasting-dependent increase in biliary E(2) may be reflective of the increased serum E(2) in fasted animals, because each 1 pg/mL increase in serum E(2) increased bile E(2) by 0.86 +/- 0.12 pg/mL, independent of nutrition (P = 0.002). Our results demonstrate that the rate of clearance of E(2) is decreased during nutritional restriction. Additionally, these data indicate that altered SHBG expression, enterohepatic recirculation, or both are involved in the decreased E(2) clearance during dietary restriction.

Estimation and prediction with HIV-treatment interruption data.

We consider longitudinal clinical data for HIV patients undergoing treatment interruptions. We use a nonlinear dynamical mathematical model in attempts to fit individual patient data. A statistically-based censored data method is combined with inverse problem techniques to estimate dynamic parameters. The predictive capabilities of this approach are demonstrated by comparing simulations based on estimation of parameters using only half of the longitudinal observations to the full longitudinal data sets.

Using specific antisera to neutralize ACTH in sturgeon: a method for manipulating the interrenal response during stress.

Interrenal function and the magnitude of the stress response were assessed in green sturgeon (Acipenser medirostris) passively immunized with antisera directed against adrenocorticotropic hormone (ACTH). The nucleotide sequence encoding ACTH was determined using reverse transcriptase polymerase chain reaction (RT-PCR). We identified two isoforms of ACTH that differ at a single site (position 26) in the 39 AA peptide. Both forms of green sturgeon ACTH (gsACTH1-39) display 100% homology with both sequences of white sturgeon ACTH (wsACTH1-39). The N-terminal portion of gsACTH also shares absolute identity with the comparable portion of human ACTH (hACTH). However, we identified considerable sequence divergence in the C-terminal domain between gsACTH and hACTH. Species-specific anti-ACTH sera were generated by vaccinating sheep against the C-terminal portion of gsACTH (gsACTH26-39). The peptide was covalently linked to a carrier protein (keyhole-limpet-hemocyanin [KLH]) to further enhance its immunogenicity. The anti-gsACTH sera recognized gsACTH1-39 and the immunogenic peptide (gsACTH26-39), but did not interact with hACTH1-39. To assess the impact of the antisera, fish were passively immunized with anti-gsACTH26-39 sera or anti-KLH sera and challenged with a hACTH1-39 injection on day 1 followed by a 1-min air emersion stressor on day 2. The magnitude and duration of the secretory response induced by hACTH did not differ (P > .05) between groups. Conversely, the magnitude of cortisol secretion induced by air emersion was significantly attenuated (P < .05) in fish passively immunized against gsACTH26-39. Collectively, these data demonstrate that the targeted antisera used in this study can discriminate between mammalian and green sturgeon ACTH and moderate the in vivo response to a stressor.

Gonadotrope function in ovariectomized ewes actively immunized against gonadotropin-releasing hormone (GnRH).

The gonadotrope cells of the ovine anterior pituitary were insulated from hypothalamic inputs by imposing an immunologic barrier generated by active immunization of ovariectomized ewes against gonadotropin-releasing hormone (GnRH) conjugated to keyhole limpet hemocyanin (KLH) through a p-aminophenylacetic acid bridge. All GnRH-KLH animals immunized developed titers of anti-GnRH that exceeded 1:5000. The antisera were specific for GnRH and cross-reacted with GnRH agonists modified in position 10 to an extent that was less than 0.01%. Ewes actively immunized against GnRH-KLH displayed levels of basal and GnRH agonist-induced gonadotropin secretion that were markedly lower (p less than 0.05) than comparable parameters in ewes actively immunized against KLH. In contrast, basal and thyrotropin-releasing hormone (TRH)-induced prolactin (PRL) secretion were not compromised by active immunization. Immunization against the GnRH-KLH conjugate, but not KLH alone, prevented expression of the positive feedback response to exogenous estradiol (E2). Pituitary stores of immunoactive luteinizing hormone (LH) and follicle-stimulating hormone (FSH) were significantly (p less than 0.001) reduced in ewes immunized against GnRH-KLH but stores of PRL were not affected by such immunization. Further, the biopotency of the residual LH stores in tissue of animals from the anti-GnRH group was significantly (p less than 0.05) lower than LH biopotency in anti-KLH animals. Serum levels of LH in anti-GnRH ewes were restored by circhoral administration of a GnRH agonist that did not cross-react with the antisera generated. Pulsatile delivery of GnRH agonist in anti-GnRH ewes significantly (p less than 0.05) elevated serum LH within 48 h and reestablished LH levels comparable to anti-KLH ewes within 6 days of treatment.(ABSTRACT TRUNCATED AT 250 WORDS)

Secretory dynamics of bioactive and immunoactive LH during the oestrous cycle of the sheep.

Blood samples were collected every 15 min for 6 h during the follicular (1 day before oestrus), and early (Days +1 to +3), mid- (Days +4 to +8), and full (Days +9 to +14) luteal phases of the oestrous cycle. Serum concentrations of immunoactive LH were measured by radioimmunoassay. The biological activity of serum LH was determined by an in-vitro bioassay that uses LH-induced testosterone production from mouse interstitial cells as an endpoint. Only ovine and bovine LH and hCG had appreciable activity in this bioassay. The temporal pattern of secretion of bioactive LH paralleled the secretory pattern of immunoactive LH at all stages of the ovine oestrous cycle. However, the secretory pattern itself varied regularly through the oestrous cycle. The frequency of secretory excursions of LH was highest during the follicular phase (6.2 +/- 0.9 pulses/6 h) and was progressively reduced through the luteal phase (1.1 +/- 0.1 pulses/6 h during full luteal phase). Conversely, amplitude of secretory excursions of immunoactive LH was low during the follicular phase (0.79 +/- 0.08 ng/ml) and significantly (P less than 0.05) increased during the mid- and full luteal phases (1.49 +/- 0.10 and 2.37 +/- 0.20 ng/ml, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)