Expression of TRPM8 in sheep reproductive tissues and brain areas important for the expression of reproductive behavior.

The Transient Receptor Potential Melastatin 8 (TRPM8) channel is a temperature-sensitive, calcium permeable ion channel and purported testosterone receptor. To determine how the hormone environment influences the expression of TRPM8 in gonadal tissue and areas of the brain important for reproduction, tissue from western white-faced cross-bred ewes, rams, and gonadectomized males (wethers; n = 6 per group) approximately 6 mo of age were collected. TRPM8 mRNA expression was greater (P = 0.01) in prostate of rams than wethers. Testes had greater (P = 0.004) expression of TRPM8 mRNA than the ovary. Differences in protein expression was similar with the testes having greater (P = 0.007) TRPM8 protein than the ovary. Protein expression did not differ (P = 0.6) in the prostate due to presence (ram) or absence (wether) of the testes. In the brain, TRPM8 varied in the amygdala with rams tending (P = 0.07) to express more mRNA which was reflected in greater (P = 0.04) number of neurons staining positive for TRPM8 in the central amygdala. Differences among ewes and wethers were not detected. This pattern was not observed (P ≥ 0.16) in the hypothalamus or olfactory bulb. To determine if TRPM8 was associated with the expression of ram sexual behavior, brains from rams categorized as high (n = 4) or low (n = 3) sexual activity were collected and blocked. Presence of TRPM8 channels was verified in the amygdala and hypothalamus of rams but was absent in the ventral tegmental area. Numbers of neurons staining positive for TRPM8 did not differ by expression of sexual behavior (P ≥ 0.2) in any area quantified. While expression of TRPM8 is more robust in tissues from intact males, expression of the channel does not appear to be important in the expression of sexual behavior.

Divining responder populations from survival data.

BACKGROUND: Biomarkers that predict treatment response are the foundation of precision medicine in clinical decision-making and have the potential to significantly improve the efficiency of clinical trials. Such biomarkers may be identified before clinical testing but many trials enroll unselected populations. We hypothesized that time-varying treatment effects in unselected trials may result from identifiable responder subpopulations that may have associated biomarkers. MATERIALS AND METHODS: We first simulated scenarios of clinical trials with biomarker populations of varying prevalence and prognostic and predictive associations to illustrate the impact of subgroup-specific effects on overall population estimates. To show a real-world example of time-dependent treatment effects resulting from a prognostic and predictive biomarker, we re-analyzed data from a published clinical trial (RTOG, Radiation Therapy Oncology Group, 9402). We then demonstrated a quantitative framework to fit survival data from clinical trials using statistical models incorporating known estimates of biomarker prevalence and prognostic value to prioritize predictive biomarker hypotheses. RESULTS: Our simulation studies demonstrate how biomarker subgroups that are both predictive and prognostic can manifest as time-dependent treatment effects in overall populations. RTOG 9402 provides a representative example where 1p/19q co-deletion and IDH mutation biomarker-specific effects led to time-varying treatment effects and a considerable deviation from proportional hazards in the overall trial population. Finally, using biomarker data from The Cancer Genome Atlas, we were able to generate statistical models that correctly identified and prioritized a commonly used biomarker through retrospective analysis of published clinical trial data. CONCLUSIONS: Biomarkers that are both predictive and prognostic can result in characteristic changes in survival results. Retrospectively analyzing survival data from clinical trials may highlight potential indications for which an underlying predictive biomarker may be found.

Effects of maternal obesity on maternal and fetal plasma concentrations of adiponectin and expression of adiponectin and its receptor genes in cotyledonary and adipose tissues at mid- and late-gestation in sheep.

Adiponectin potentially influences fetal weight by altering insulin signaling and trans-placental amino acid and glucose transporters. The objective of this study was to determine how maternal obesity influences maternal and fetal plasma concentrations of adiponectin, expression of fetal adiponectin, its receptors, and adipogenic genes at mid- and late-gestation. Blood samples and tissues were collected from obese and control multiparous pregnant ewes at day 75 or 135 of gestation. Although day of gestation or maternal obesity did not influence (P > 0.6) maternal plasma concentrations of adiponectin, fetal weight was increased (P < 0.001) and adiponectin tended to decrease (P = 0.10) at mid-gestation in fetuses from obese ewes. Differences were not apparent at late-gestation (P > 0.70). Relative abundance of adiponectin (P = 0.01), AdipoR2 (P = 0.04) and PPARγ (P = 0.01) mRNA was less at mid-gestation in fetal adipose tissue from obese mothers. By late gestation, maternal obesity tended to associated with a decrease in relative abundance of adiponectin (P = 0.09) and SREBF1 (P = 0.10) mRNA in fetal adipose tissue. Maternal obesity did not influence (P ≥ 0.20) the relative abundance of adiponectin, AdipoR1 and AdipoR2 mRNA in cotyledonary tissue at mid or late- gestation. In conclusion, maternal obesity in sheep influences relative abundance of fetal adipose tissue mRNA for adiponectin and adipogenic, as well as plasma concentrations of total adiponectin. Although adiposity in pregnant ewes did not influence maternal adiponectin, maternal obesity potentially influenced fetal adipogenesis by altering the abundance of adiponectin, PPARγ and SREBF1 mRNA in fetal adipose tissue.

Depo-Provera increases neural activity in the central amygdala of reindeer bulls.

Reindeer bulls are difficult to manage and dangerous to handlers during the rutting period. Progesterone agonists have been used anecdotally in the field to favorably influence behavior, but effects on reproductive signaling have not been determined. The objective of this study was to determine the effects of Depo-Provera (medroxyprogesterone acetate) on neural activity in the amygdala of reindeer bulls in the early (n = 4) and full (n = 4) rut. Treated bulls (n = 4) were injected with a single injection of Depo-Provera (400 mg i.m.) approximately 2 wk before rut was initiated. Control bulls were untreated. Bulls were exsanguinated and brains collected. Neural activity in the amygdala was determined using c-fos immunohistochemistry. Neural activity did not differ by treatment (P ≥ 0.5), collection period (P ≥ 0.5), or their interaction (P ≥ 0.3) in the medial and cortical amygdala nuclei. A treatment × time interaction (P = 0.009) was observed in the central amygdala. The amygdala nuclei are interconnected allowing for integration of sensory stimuli with a direct connection between the medial amygdala and the olfactory bulb. The central amygdala is responsible for alerting, fear, and initiating a state of arousal towards nonspecific stimuli in the surrounding environment. In wildlife, the central amygdala has a role in recognizing threats in the environment such as predators. During the rut, bulls normally have a decreased sense of fear and elevated aggressive behavior with Depo-Provera treatment seemly able to diminish that aggression. Although it is unlikely that this observed change in neural activity fully explains the decreased aggressive behavior noted in bulls treated with Depo-Provera, neural networks of aggression include the amygdala. It is possible that further changes in c-fos activity will be noted in other areas of the brain known to be necessary for processing social cues. Bulls treated with Depo-Provera maintain sexual interest and have offspring. Depo-Prevera increases the neural activity within the central amygdala and may partially account for their altered aggressive behavior during the rut.

Tyrosine hydroxylase in the ventral tegmental area of rams with high or low libido-A role for dopamine.

Dopamine synthesis in the ventral tegmental area (VTA) is necessary for the reinforcement of sexual behavior. The objective of this study determined if sexual stimuli initiates reward, and whether reward is attenuated in sexually inactive rams. Sexually active rams were exposed to urine from estrous (n=4) or ovariectomized (n=3) ewes with inactive rams (n=3) exposed to urine from estrous ewes. Following exposure, rams were exsanguinated and brains perfused. Alternating sections of the VTA were stained for Fos related antigens (FRA), tyrosine hydroxylase, and dopamine beta-hydroxylase activity. Forebrain tissue, mid-sagittal ventral to the anterior corpus callosum, was stained for dopamine D2 receptors. Concentrations of cortisol was determined prior to and following exposure. Exposure to ovariectomized-ewe urine in sexually active rams did not influence (P=0.6) FRA expression, but fewer (P<0.05) neurons were positive for tyrosine hydroxylase in the VTA. Sexually inactive rams had fewer (P<0.05) FRA and tyrosine hydroxylase positive neurons in the VTA than sexually active rams following exposure to estrous ewe urine. VTA neurons staining positive for dopamine beta-hydroxylase did not differ by sexual activity (P=0.44) or urine exposure (P=0.07). Exposure to stimulus did not influence (P=0.46) numbers of forebrain neurons staining positive for dopamine D2 receptors in sexually active rams, but fewer (P=0.04) neurons stain positive in inactive rams. Serum concentrations of cortisol did not differ (P≥0.52) among rams prior to or following stimulus. In conclusion sexual inactivity is unlikely due to stress, but may be partially a result of decreased tyrosine hydroxylase and/or the response to dopamine.

Whole brain radiotherapy after stereotactic radiosurgery or surgical resection among patients with one to three brain metastases and favorable prognoses: a secondary analysis of EORTC 22952-26001.

BACKGROUND: The absence of a survival benefit for whole brain radiotherapy (WBRT) among randomized trials has been attributed to a competing risk of death from extracranial disease. We re-analyzed EORTC 22952 to assess the impact of WBRT on survival for patients with controlled extracranial disease or favorable prognoses. PATIENTS AND METHODS: We utilized Cox regression, landmark analysis, and the Kaplan-Meier method to evaluate the impact of WBRT on survival accounting for (i) extracranial progression as a time-dependent covariate in all patients and (ii) diagnosis-specific graded prognostic assessment (GPA) score in patients with primary non-small-cell lung cancer (NSCLC). RESULTS: A total of 329 patients treated per-protocol were included for analysis with a median follow up of 26 months. One hundred and fifteen (35%) patients had no extracranial progression; 70 (21%) patients had progression <90 days, 65 (20%) between 90 and 180 days, and 79 (24%) patients >180 days from randomization. There was no difference in the model-based risk of death in the WBRT group before [hazard ratio (HR) (95% CI)=0.70 (0.45-1.11), P = 0.133), or after [HR (95% CI)=1.20 (0.89-1.61), P = 0.214] extracranial progression. Among 177 patients with NSCLC, 175 had data available for GPA calculation. There was no significant survival benefit to WBRT among NSCLC patients with favorable GPA scores [HR (95% CI)=1.10 (0.68-1.79)] or unfavorable GPA scores [HR (95% CI)=1.11 (0.71-1.76)]. CONCLUSIONS: Among patients with limited extracranial disease and one to three brain metastases at enrollment, we found no significant survival benefit to WBRT among NSCLC patients with favorable GPA scores or patients with any histology and controlled extracranial disease status. This exploratory analysis of phase III data supports the practice of omitting WBRT for patients with limited brain metastases undergoing SRS and close surveillance. CLINICAL TRIALS NUMBER: NCT00002899.

Fos Expression in the Olfactory Pathway of High- and Low-Sexually Performing Rams Exposed to Urine from Estrous or Ovariectomized Ewes.

Exposure to estrous ewe urine stimulates investigation and mounting activity in sexually active but not sexually inactive rams. It was hypothesized sexual indifference may result from an inability to detect olfactory cues or an interruption of the pathway from detection of the olfactory stimulus to the motor response. Sexually active (n=4) and inactive (n=3) rams were exposed to urine from estrous ewes. An additional group of sexually active rams (n=3) were exposed to urine from ovariectomized ewes. Rams were exsanguinated following 1 h of exposure to stimulus. Neural activity was determined in tissues of interest by the presence of fos and fos-related proteins detected by immunohistochemistry procedures. Sexually active rams exposed to urine from ovariectomized ewes had more (P ≤ 0.05) fos-positive cells in the olfactory bulb, but fewer (P = 0.03) fos-positive cells in the cortical amygdala compared to sexually active rams exposed to urine from estrous ewes. Sexually inactive rams had similar (P ≥ 0.13) numbers of fos positive neurons in the olfactory bulb and medial amygdala but fewer (P ≤ 0.04) in the central amygdala, bed nucleus of the stria terminalis and the medial preoptic area compared to sexually active rams exposed to urine from estrous ewes. Sexual inactivity was not associated with decreased hypothalamic function since fos activity was similar (P ≥ 0.14) among groups in the suprachiasmatic and ventral medial nucleus. Sexual inactivity is not likely due to an impaired ability to detect or process olfactory stimuli by the main olfactory bulb and medial-cortical amygdala. Sexually inactive rams may have reduced attentiveness to sexual stimuli and/or decreased responsiveness of regions in the brain which regulate reproductive behaviors.

Sunlight exposure increases vitamin D sufficiency in growing pigs fed a diet formulated to exceed requirements.

Traditional confinement practices limit exposure to sunlight and vitamin D synthesis, and vitamin insufficiency occurs even with dietary supplementation. The aim of this study was to determine the effect of limited sun exposure on serum concentration of vitamin D and the expression of vitamin D synthesizing enzymes in the liver and kidney of pigs on a vitamin D sufficient diet. White-pigmented grower pigs (29.7 ± 2.3 kg) fed 15% CP diet ad libitum providing >1,200 IU vitamin D3/kg of feed were exposed to sunlight for 1 h each day at solar noon for 14 d at the spring equinox (March pigs, n = 10) or summer solstice (June pigs, n = 5) and again before slaughter in June (March pigs) and September (June pigs). Blood for the analysis of 25(OH)D was collected before and after sunlight exposure. Traditionally housed pigs served as controls. After initial sun exposure, blood samples were collected from June pigs daily for 5 d and weekly for 8 wk to determine vitamin D3 and 25(OH)D decay, respectively. Kidney and liver samples were collected from the June pigs at slaughter after sun exposure for analysis of messenger RNA expression of vitamin D binding protein and synthesizing/degrading enzymes. Average daily gain (ADG) was not influenced (P > 0.5) by sunlight exposure. June pigs had fewer days on feed, lower (P = 0.003) ADG and were slaughtered at a lighter (P < 0.001) weight. Exposure to sunlight increased (P < 0.001) 25(OH) vitamin D for all pigs. March pigs, obtained from a Midwest producer, had lower (P < 0.001) concentration of 25(OH)D than June pigs born on-farm. Initial sunlight exposure increased serum concentration of 25(OH)D in March pigs by 200% and June pigs by 67%. Serum concentration of vitamin D3 was decreased (P < 0.05) by 72 h with 25(OH)D decreased (P < 0.05) by wk 4 after exposure. Expression of vitamin D binding protein, vitamin D synthesizing CYP2R1, CYP27A1, CYP2D25, or degrading enzyme CYP24A1 were not influenced (P ≥ 0.19) by sunlight exposure. Expression of CYP27B1 was decreased (P = 0.04) in the kidney but tended to be increased (P = 0.06) in the liver after sun exposure. These results suggest limited sun exposure can efficiently increase serum concentration of vitamin D in growing pigs with varying levels of vitamin sufficiency. The lack of major changes in vitamin synthesizing enzymes suggests the 14-d exposure period did not saturate the capacity of slaughter-weight pigs to synthesize vitamin D.

50 years of the Wyoming ram test: How sheep have changed.

Production characteristics of white-faced rams have been systematically evaluated over a 140-d test in Wyoming since 1961. Individual test records ( = 4,240) from rams on test were analyzed to determine change over the past 52 yr. Although rams on test are not older, weight on and off test has increased ( < 0.001) since 1961. Weight off test increased 22.7 kg and contributed to an increase ( < 0.001) in clean fleece weight. Rate of gain ( < 0.001) almost doubled over this 50-yr period. Growth efficiency improved from 0.23 ± 0.01 kg/d from 1961 to 1966 to 0.39 ± 0.01 kg/d from 2008 to 2013. Cubic, rather than linear, effects better explain the change in growth characteristics, suggesting a plateau or tapering of these traits. Wool characteristics remain an important component of the test index, and despite increases in body size and gain, wool diameter was unchanged ( > 0.15). Average daily gain correlated ( > 0.67; < 0.001) with lamb and feeder lamb price, with the strongest correlation at a 2-yr ( > 0.76) time lag. U.S. sheep inventory was negatively correlated ( > -0.72; < 0.001) with sheep price and ADG, with the greatest correlation at no time lag. Wool price 0, 2, or 5 yr prior did not correlate ( < 0.1; ≥ 0.5) with spinning count. Influences on white-faced ram selection appear to have largely impacted growth traits while avoiding negative impacts on wool quality.

Effects of progesterone and RU486 on the development and expression of adult male sexual behaviour and gene expression in the amygdala and preoptic area of the hypothalamus.

The number of progesterone receptors is greater in the male than female neonatal rat hypothalamus. The aims of the present study were to determine developmental effects of progesterone on the expression of adult male sexual behaviour and whether changes in behaviour were reflected by altered gene expression within the hypothalamic preoptic area (POA) or medial amygdala. Male rats were treated with progesterone (40 µg kg(-1), i.p.), the progesterone receptor antagonist RU486 (40 µg kg(-1), i.p.) or an equal volume of vehicle (10% ethanol, 90% corn oil) on postnatal Days 1-5. Treatment with either progesterone or RU486 inhibited (P ≤ 0.07) the initial expression of consummatory sexual behaviour at 10.5 weeks of age without influencing growth or serum concentrations of testosterone. Sexual interest, as measured by latency to exhibiting mounting behaviour or the number of mounts achieved, was not influenced by treatment with either progesterone or RU486. The effects of treatment with progesterone or RU486 on sexual behaviour were diminished by experience. Microarray analysis of the POA indicated 61 genes that were upregulated and 49 that were downregulated (P ≤ 0.01) following RU486 treatment of male rats. However, the altered expression of selected genes was not confirmed by real-time reverse transcription-polymerase chain reaction. The expression of targeted genes within the amygdala was not influenced by treatment with either progesterone or RU486. Neonatal treatment with RU486, but not progesterone, decreased testes weight (P=0.02) without affecting testes morphology. The results indicate that altering the progesterone environment during a critical developmental period affects the expression of behaviour, but that changes in behaviour are not mirrored by the altered expression of selected genes.

Renin mRNA is upregulated in testes and testicular cells in response to treatment with aflatoxin B1.

Aflatoxin B(1) (AFB(1)) has been shown to affect fertility in many species; however, the exact molecular mechanisms associated with the disruption are not known. Our objectives were to determine changes in testicular gene expression due to exposure to AFB(1) and to investigate which cell types were affected by treatment with AFB(1). Male mice 4 wk of age were administered a daily placebo (control; N = 9) or 50 µg/kg AFB(1) (AFB(1) treated; N = 10) daily for 45 days. Males were then mated to four females each for 8 days. Male mice were characterized as being "Tolerant" (N = 3) or "Intolerant" (N = 3) to the effects of AFB(1) based on positive terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining in the testes and the number of pups sired. Tolerant males produced a similar average number of fetuses (mean ± SEM) (12.5 ± 1.2) per male as selected control males (13.4 ± 1.2), but more fetuses (P = 0.01) than Intolerant males (7.6 ± 1.2). The number of terminal deoxynucleotidyl transferase dUTP nick end labeling-positive cells in Intolerant males tended to be (P = 0.10) greater (136.5 ± 27.2) than in Tolerant (55.0 ± 22.2) and selected control (54.3 ± 22.2) males. Affymetrix microarray (Sunnyvale, CA, USA) analysis revealed differential expression (P < 0.05) of 193 extra cellular space and signaling genes, 49 signal transduction genes, 45 immune regulation genes, and 230 cell differentiation genes in the testis. Renin was commonly represented amongst many clusters and was chosen for further analyses. Upregulation (P < 0.001) of Renin in Tolerant mice (N = 3) compared with Intolerant mice (N = 3) was confirmed by real-time reverse transcription polymerase chain reaction (RT-PCR) (P = 0.05). This upregulation (P = 0.01) was also observed in representative AFB(1) treated males (N = 8) compared with control males (N = 8) selected for real-time reverse transcription polymerase chain reaction analysis. Spermatogonia cultured in vitro and treated with 0, 5, 10, or 20 µg/mL AFB(1) (N = 6 per treatment) resulted in a 10-fold upregulation (P = 0.01) of Renin message at the 20 µg/mL level, whereas Leydig tumor cells showed similar differences (P = 0.03) in message for Renin in treated (10 and 20 µg/mL) versus control cell cultures. Based on these results, we inferred a role for Renin at the molecular level in the response to the adverse effects of AFB(1) in male mice.

Hypothalamic expression of genes for appetite regulators and estrogen α, estrogen ß and leptin receptors in obese dams and their fetuses.

Under- and over-nutrition during gestation may influence fetal hypothalamic development resulting in individuals predisposed to adverse health effects. This study examined fetuses from obese and control ewes to determine whether dam obesity alters hypothalamic expression of fetal appetite regulatory genes. A second objective was to contrast the expression of appetite regulatory genes in ewes that become the most obese to those that remained in moderate body condition on the same energy-rich diet. Multiparous, western white-faced ewes were weighed and individually fed 100% (control) or 150% (obese) of National Research Council requirements from day 60 before mating until day 75 of gestation. At day 75 of gestation, fetuses were collected and weighed. Hypothalamic tissue from fetal lambs and dams was collected and frozen for mRNA extraction. Dam obesity (P ≥ 0.16), fetal sex (P ≥ 0.44) or their interaction (P ≥ 0.42) did not affect the relative expression of fetal hypothalamic regulators of appetite, including neuropeptide Y, agouti-related protein, pro-opiomelanocortin, cocaine- and amphetamine-regulated transcript and receptors for leptin. Maternal obesity at day 75 of gestation in ewes did not affect developmental mechanisms responsible for the expression of fetal appetite regulatory genes and would not be expected to predispose offspring to adult-onset obesity through disrupted appetite regulation at this developmental time point. In the ewe, appetite regulatory genes did not differ (P > 0.20) with ewe adiposity; however, expression of estrogen receptor α, but not ß (P = 0.37), in the medial basal hypothalamus was greater (P = 0.04) in obese than in control ewes.

Effects of feeding high-linoleate safflower seeds on postpartum reproduction in beef cows.

Two experiments were conducted to evaluate reproductive responses to supplemental high-linoleate safflower seeds in postpartum beef cows. In Exp. 1, 18 primiparous, crossbred beef cows (411 +/- 24.3 kg of BW) were fed Foxtail millet hay starting 1 d postpartum at 1.68% of BW (DM basis) and a low-fat control (control: 63.7% cracked corn, 33.4% safflower seed meal, and 2.9% liquid molasses; DM basis) at 0.35% of BW (n = 9) or a supplement (linoleate) containing 95.3% cracked high-linoleate (79% 18:2n-6) safflower seeds and 4.7% liquid molasses (DM basis) at 0.23% of BW (n = 9). Beginning 1 d postpartum, blood was collected every 3 d for sera. Cows were slaughtered at 37 +/- 3 d postpartum for collection of hypothalami, anterior pituitary glands, liver, ovarian follicles, and uterine tissue. By 37 +/- 3 d postpartum, dietary treatment did not influence ovarian follicular development (P >or= 0.17), hypophyseal concentrations of LH (P = 0.14), or concentrations of IGF-I in liver (P = 0.15). In contrast, anterior pituitary glands from linoleate cows contained more FSH (P = 0.02) than control cows and linoleate cows had less IGF-I in the medial basal hypothalamus (P = 0.05), preoptic area (P = 0.06), and in follicular fluid (P

Influence of short-term fasting during the luteal phase of the estrous cycle on ovarian follicular development during the ensuing proestrus of ewes.

Short-term fasting of mature ewes during diestrus results in increased serum concentrations of progesterone and a delayed pre-ovulatory surge release of LH. To determine if these changes in reproductive hormones influence subsequent follicular development, mature ewes observed in estrus were assigned randomly to control (n=10) or fasted (n=15) groups. Control ewes had ad libitum access to feed, whereas fasted ewes were not fed from day 7 through 11 of their estrous cycle. Daily blood samples were collected from control and fasted ewes throughout the fasting period. Fasting increased (P<0.001) serum concentrations of progesterone (4.4 ng/mL versus 2.7 ng/mL [+/-0.3]). On day 12, all ewes were treated with 10mg of PGF(2alpha) and fasted ewes were returned to ad libitum feed. Ovaries were collected from ewes (n=5 each group) at 0 and 72 h following PGF(2alpha) in control and 0, 72 and 96 h in fasted ewes. Ovaries were weighed and small (< or =2mm), medium (3-4mm), and large (> or =5mm) follicles were enumerated. Total numbers of follicles were less (P<0.001) in fasted than fed ewes (14.6 versus 30.2 [+/-2.2]) at 0 h, but did not differ (P=0.9) when numbers of follicles were compared at similar times before the anticipated LH surge (i.e., at 72 h versus 96 h in control and fasted ewes, respectively). Within follicular size class, numbers of small and medium follicles were decreased (P=0.04) at 0 h in fasted ewes. Numbers of large follicles did not differ (P=1.0) between groups. Although numbers of small and medium ovarian follicles in fasted ewes recovered by 96 h to values comparable to fed ewes at 72 h following PGF(2alpha), serum concentrations of estradiol 17beta (P=0.08) and FSH (P=0.06) tended to be decreased in fasted ewes before the anticipated surge release of LH. Pituitary content of LH and FSH also tended to be lower (P< or =0.09) at 96 h in fasted ewes than at 72 h in control ewes, but did not differ (P> or =0.4) at hour 0 following PGF(2alpha). Hypothalamic and stalk median eminence contents of GnRH were not influenced (P> or =0.2) by fasting at any time period. Fasting during the luteal phase perturbs gonadotropin secretion and may influence fertility by causing a delay in ovarian follicle development.

Effects of postpartum dietary fat and body condition score at parturition on plasma, adipose tissue, and milk fatty acid composition of lactating beef cows.

Two experiments were conducted with lactating Angus x Gelbvieh beef cows to determine the effects of postpartum lipid supplementation, BCS at parturition, and day of lactation on fatty acid profiles in plasma, adipose tissue, and milk. In Exp. 1, 36 pri-miparous cows (488 +/- 10 kg of initial BW; 5.5 +/- 0.02 initial BCS) were given ad libitum access to hay and assigned randomly to a low-fat (control) supplement or supplements with cracked, high-linoleate safflower seeds (linoleate) or cracked, high-oleate safflower seeds (oleate) from d 3 to 90 of lactation. Diets were formulated to be isonitrogenous and isocaloric; safflower seed diets provided 5% of DMI as fat. Plasma and milk samples were collected on d 30, 60, and 90 of lactation. Adipose tissue biopsies were collected near the tail-head region of cows on d 45 and 90 of lactation. In Exp. 2, 3-yr-old cows achieving a BCS of 4 +/- 0.07 (479 +/- 36 kg of BW) or 6 +/- 0.07 (580 +/- 53 kg of BW) at parturition were used in a 2-yr experiment (n = 36/yr). Beginning 3 d postpartum through d 61 of lactation, cows were fed diets similar to those of Exp. 1. Adipose tissue and milk samples were collected on d 30 and 60, and plasma was collected on d 31 and 61 of lactation. Responses to postpartum dietary treatment were comparable in both experiments. Cows fed linoleate and oleate had greater (P < 0.001) total fatty acid concentrations in plasma than cows fed control. Except for 15:1, milk fatty acids with <18 carbons were greatest (P < or = 0.01) for cows fed control, whereas milk from cows fed linoleate had the greatest (P < or = 0.02) 18:1trans-11, 18:2n-6, and cis-9, trans-11 CLA. Milk from cows fed oleate had the greatest (P < 0.001) 18:1cis-9. In Exp. 1, total fatty acid concentrations in adipose tissue samples decreased at d 90 compared with d 45 of lactation, but the fatty acid profile of cow adipose tissue was not affected (P = 0.14 to 0.80) by dietary treatment. In Exp. 2, the percentage of cis-9, trans-11 CLA in adipose tissue of cows with a BCS of 6 decreased (P = 0.001) from d 30 to 60 of lactation. Plasma and milk fatty acid composition reflected alterations in postpartum diet. Less medium-chain fatty acids and more 18-carbon fatty acids in milk were indicative of reduced de novo fatty acid synthesis in the mammary gland of beef cows fed lipid supplements; however, the metabolic demands of lactation prevented the deposition of exogenously derived fatty acids in adipose tissue through d 90 of lactation.

The incidence of genotypes at codon 171 of the prion protein gene (PRNP) in five breeds of sheep and production traits of ewes associated with those genotypes.

Scrapie is one of several transmissible spongiform encephalopathies of livestock. Disease susceptibility is linked to polymorphisms in the normal prion protein gene that encodes the mammalian prion precursor. Codon 171 of this gene is a major determinant of scrapie susceptibility. Selection for arginine (R) at codon 171 is encouraged by the USDA to decrease the incidence of scrapie. Objectives of this study were to determine the frequency of R allele variants at codon 171 in a sample of sheep from five breeds (Columbia, Hampshire, Rambouillet, Suffolk, and Targhee) and western white-faced commercial ewes and to determine whether the R allele is associated with ewe and lamb production traits. Genotyping was performed on 532 ewes and 901 lambs from the University of Wyoming flock, in addition to 820 rams from 52 sheep producers from Wyoming and surrounding areas, using a DNA mismatch assay that discriminated the R allele from others at codon 171. Genotyping was performed by DNA sequencing on 127 rams representing all breeds, except Hampshire from the USDA Sheep Experiment Station at Dubois, ID. The 171R allele was found in all five breeds and in the commercial western white-faced ewes. Genotype frequencies varied (P < 0.001) by breed in ewe and ram populations. Influence of R-allele frequency on ewe lambing records and individual lamb records was analyzed for Columbia (62, 161, 121), Hampshire (89, 193, 162), Rambouillet (87, 179, 133), Suffolk (67, 178, 161), and commercial sheep (227, 463, 324) for numbers of ewes, total number of ewe production records, and individual lamb records, respectively. Suffolk ewes without the R allele (non-R/non-R) gave birth to more (P or= 0.08) by ewe genotype. Lamb birth and weaning weights were not influenced (P >or= 0.12) by lamb genotype in any of the breeds or in the commercial flock. In this population, ultimate lamb production was only influenced by genotype at codon 171 in the Suffolk flock.

Effects of feed restriction on reproductive and metabolic hormones in ewes.

The goal of this study was to determine the effects of short-term feed withdrawal on reproductive and metabolic hormones during the luteal phase of the estrous cycle in mature ewes. Mature ewes observed in estrus were assigned randomly to control and fasted groups (n = 10 per group Trials 1 and 2). For Trials 1 and 2, control ewes had ad libitum access to feed, whereas fasted ewes were not fed from d 7 through 11 of their estrous cycle; on d 12, all ewes were treated with 10 mg of PGF2alpha, and fasted ewes were gvien ad libitum access to feed. For Trial 1, blood samples were collected daily through fasting and at 2-h intervals following PGF2alpha for 72 h. Serum concentrations of insulin (P < or = 0.002) and IGF-I (P < or = 0.01), but not GH (P > or = 0.60), were decreased during fasting compared with fed ewes. Serum concentrations of 29 (P = 0.02) and 34 kDa (P = 0.04) IGFBP were greater in fasted ewes at 96 h after initiation of fasting than in control ewes. Two control and four fasted ewes in Trial 1 did not exhibit a preovulatory surge release of LH by 72 h. Therefore, Trial 2 was conducted so that the timing of the LH surge could be predicted following the collection of blood samples at 2-h intervals for 112 h and then at 6-h intervals until 178 h following PGF2alpha administration and realimentation. The magnitude of the preovulatory LH surge in Trial 2 was decreased (P = 0.009) and delayed (P = 0.04), and serum concentrations of estradiol were diminished (P < or = 0.03) 12 h before the LH surge in fasted ewes. Ovulation rates were not influenced (P > or = 0.32) by fasting in Trials 1 and 2. Serum concentrations of progesterone in both Trials 1 and 2 were, however, greater (P < 0.001) in fasted than in control ewes. A third trial with ovariectomized ewes was conducted to determine whether the increased serum concentrations of progesterone observed in fasted ewes during Trials 1 and 2 were ovarian-derived. Ovariectomized ewes were implanted with progesterone-containing intravaginal implants and allotted to control (n = 5) or fasted (n = 5) treatment groups and fed as described for Trials 1 and 2. Similar to intact ewes, serum concentrations of progesterone were approximately twofold greater (P < 0.001) in fasted than in control implanted ovariectomized ewes. In summary, feed withdrawal for 5 d during the luteal phase of the estrous cycle increased serum concentrations of progesterone and evoked endocrine changes that could perturb the subsequent estrous cycle.

Effects of supplementation with high linoleic or oleic cracked safflower seeds on postpartum reproduction and calf performance of primiparous beef heifers.

Primiparous Angus x Gelbvieh (n = 36) rotationally crossed beef cows (initial BW = 487.9 +/- 10.5 kg, body condition score = 5.5 +/- 0.02) were utilized to determine effects of supplemental safflower seeds high in linoleic (76% 18:2) or oleic (72% 18:1) acid on cow BW change, body condition score, milk production and composition, calf weight gain, cow serum metabolites, and metabolic hormones. On d 3 postpartum, cows were randomly assigned to one of three isonitrogenous dietary supplements with equal total quantity of TDN: corn-soybean control supplement (n = 12); high-linoleate safflower seeds (n = 12); or high-oleate safflower seeds (n = 12). Safflower-seed supplements were formulated to provide 5% DMI as fat. Supplements were individually fed from d 3 postpartum through 90 d postpartum. Cows had ad libitum access to native grass hay (7.8% CP), trace-mineralized salt, and water. Date of parturition was evenly distributed across treatments with all cows calving within 14 +/- 0.8 d. There were no differences (P = 0.65) in total OM intake among treatments. Although cow BW change did not differ (P = 0.33) by treatment, supplementation influenced cow body condition score (P = 0.02) with linoleate-supple-mented cows in higher (P = 0.005) condition overall than oleate-supplemented cows (5.1 +/- 0.06 vs 4.9 +/- 0.06). Twenty-four-hour milk production did not differ (P = 0.68) among treatments. Percentage milk fat was not different at d 30; however, at d 60 and d 90 percentage milk fat was greater (P ( 0.05) in control and oleate-supplemented cows than in linoleate-supplemented cows. Calf BW gains (P = 0.27) and adjusted 205-d weights (P = 0.48) were not affected by supplement treatment. Supplementation did not influence serum concentrations of glucose (P = 0.38), NEFA (P = 0.61), GH (P = 0.29), IGF-I (P = 0.81), insulin (P = 0.26), or IGF-I binding proteins (P > or = 0.11). Days to conception did not differ (P = 0.40) among treatments. Although overall productivity of the primiparous cows and their calves was not altered by safflower-seed supplementation, differential effects were noted between supplements. Oleate supplementation increased percentage milk fat at d 60, and cow body condition score was lower than in linoleate-supplemented cows. Linoleate-supplemented cows had greater body condition scores by 90 d postpartum than either corn-soybean- or oleatesupplemented cows.