Type A allatostatins from Drosophila melanogaster and Diplotera puncata activate two Drosophila allatostatin receptors, DAR-1 and DAR-2, expressed in CHO cells.

The type-A allatostatins A (AST-A) are a group of insect peptides with a common C-terminal motif Y/FXFGL-NH(2). The existence of at least four putative type A Drosophila melanogaster ASTs (called type A drostatins or DST-As) has been predicted from the sequence of a recently cloned DST-A preprohormone [C. Lenz et al. (2000) Biochem. Biophys. Res. Commun. 273, 126-1131]. SRPYSFGL-NH(2), (DST-3A), the only DST isolated from Drosophila so far, activated the first cloned DST-A GPCR (DAR-1) [N. Birgül et al. (1999) EMBO J. 18, 5892-5900]. A newly cloned orphan Dm GPCR, which shares 47% overall and 60% transmembrane region sequence identity with DAR-1, was classified as a second putative Dm DST-A receptor (DAR-2) [C. Lenz et al. (2000) Biochem. Biophys. Res. Commun. 273, 571-577]. Although activation of DAR-2 by DSTs has been postulated, no experimental evidence for that has been presented to date. In this study, we expressed both DAR-1 and DAR-2 in CHO cells and used a GTPgammaS and a Ca(2+) mobilization assay for pharmacological evaluation of the receptors. Synthetically prepared DST-As, as well as selected Diplotera punctata (cockroach) ASTs, activated DAR-1 and DAR-2 in both functional assays indicating ligand redundancy and cross species activity. Cell pretreatment with pertussis toxin led to some differences in the nature and magnitude of signaling pathways at the DAR-1 and DAR-2 receptors, suggesting possible differential coupling to cellular effector system(s) and distinct biological functions of each receptor in vivo.

Pig weaning weight and changes in hematology and blood chemistry of sows injected with recombinant porcine somatotropin during lactation.

Weaning weight of pigs on d 18 to 21 of age was used to determine the effects of recombinant porcine somatotropin (rpST) on lactational performance of their dams. Crossbred Landrace x Yorkshire sows (n = 180, 202 to 270 kg BW) were assigned to receive daily i.m. injections of either 0, 8, or 16 mg of rpST on lactation d 7 to 20. Initially, injections of 16 mg of rpST/d caused 100% mortality (four sows); thus, a dose of 4 mg of rpST/d replaced 16 mg. Subsequently, 5 of 19 and 1 of 17 sows died in response to daily injections of 8 and 4 mg of rpST, respectively, resulting in termination of the experiment. Before termination, 48 sows (16 from each of the 0-, 4-, and 8-mg treatment groups) completed a lactation of 18 to 21 d in duration. Analysis of these data indicated that daily injection of lactating sows with rpST increased (P less than .05) weaning weight of pigs only if they were from litters that averaged greater than or equal to 2.6 kg/pig on lactation d 7. The BW of sows was decreased by rpST (P = .07). Subsequent experiments revealed 1) that rpST-induced death of lactating sows was caused from hemorrhaging of ulcers that developed at the pars esophagea, 2) that there was vacuolation of the liver and kidney of dead sows, and 3) that daily injection of 16 mg of rpST into nonlactating growing pigs (50 to 100 kg BW) for 28 d did not cause death or any observable illness in spite of 100% mortality of lactating sows after two to nine injections.

The interrelationship between crude protein and exogenous porcine somatotropin on growth, feed and carcass measurements of pigs.

In three experiments the interrelationship between dietary CP and recombinant porcine somatotropin (rpSt, i.m. daily) on ADG, feed efficiency (F/G) and carcass traits was examined in crossbred Yorkshire gilts and barrows given ad libitum access to their diets during the finishing period (55 to 110 kg BW). Pigs, blocked by BW and gender, were assigned (four/pen) within block. In Exp. 1, 140 pigs were assigned two/gender per pen to each of five pens/block and received a diet of either 12%, 18% or 24% CP (n = 2, 1 and 2 pens/block, respectively). Pigs received rpSt, either 0 or 120 micrograms/kg BW (12% and 24% CP groups) or 60 micrograms/kg BW (18% CP group). When CP was 12%, rpSt decreased ADG and increased F/G (P less than .05), whereas when CP was 18% or 24%, rpSt increased ADG and lowered F/G (P less than .05). Backfat thickness was reduced (P less than .05) by rpSt regardless of CP. In Exp. 2, 120 pigs were assigned two/gender per pen to each of five pens/block and received a diet of 24% CP. Either 0, 15, 30, 60 or 120 micrograms of rpSt/kg BW was administered to each pig. All doses of rpSt increased ADG, lowered F/G and decreased backfat thickness compared with measurements for control pigs (P less than .05). In Exp. 3, 140 pigs were assigned two/gender per pen to each of seven pens/block and received a diet of either 14%, 18% or 24% CP (n = 3, 2 and 2 pens/block, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)

Growth performance and carcass characteristics of pigs administered recombinant porcine somatotropin during 30 to 110 kilogram live weight.

To determine growth performance during and after injection of recombinant porcine somatotropin (rpSt), crossbred Yorkshire gilts and barrows (n = 54/gender, 27 to 42 kg BW) were blocked by BW and gender (n = 3 blocks/gender). Within each block, three pigs/gender were assigned randomly to each of six pens/block. A diet containing 24.8% CP was fed ad libitum. During the live weight period of 30 to 110 kg, pigs either remained as controls (one pen/block) or were injected (i.m.) daily with rpSt (120 micrograms/kg BW) during either 30 to 60, 30 to 100, 30 to 110, 60 to 100 or 60 to 110 kg BW. Thus, three gilts and three barrows in each of three pens received each treatment. Pigs were slaughtered at either 60 kg BW (1 d after rpSt injection) or 110 kg BW (1 d, 10 d or 70 d after rpSt injection). Relative to controls, pigs injected with rpSt exhibited faster and more efficient growth during the injection period (P less than .05) but slower and less efficient growth during 10 d after cessation of injection (P less than .05). Carcasses of pigs slaughtered 1 d or 10 d after rpSt injection were leaner than controls (P less than .05), but among the pigs treated with rpSt, carcasses of pigs withdrawn from rpSt for 10 d contained more fat (P less than .05) and had a lower percentage of muscle (P less than .05) than carcasses of pigs withdrawn from rpSt for 1 d. Growth and carcass measurements were similar (P greater than .05) between controls and pigs killed 70 d after rpSt injection.(ABSTRACT TRUNCATED AT 250 WORDS)

Carnivore alteration of human bone from a late prehistoric site in Illinois.

The nature and pattern of destruction of human bone by scavenging animals, probably canids, is described for 30 Oneota skeletons from a west-central Illinois cemetery dated at ca. A.D. 1300. The most frequently damaged parts of the skeleton include bones covered by relatively little soft tissue, such as the major joints of the limbs, and those in the facial, abdominal, and gluteal regions.

Response of postpartum beef cows to exogenous progestogens and gonadotropin releasing hormone.

Plasma progesterone (P4) profile and estrous detection were used during three experiments to evaluate the effects of exogenous progestogens on the life span of gonadotropin releasing hormone (GnRH)-induced corpora lutea (CL) in postpartum (pp) beef cows. Experiment 1 utilized primiparous fall-calving cows (n = 28, trial 1); and spring-calving cows (n = 29, trial 2). On d 18 to 27 pp (d 0) all cows received intravaginal devices containing either P4 or no P4 (NP) for 5 d. On d 5 the devices were removed and calves were either removed (CR) or were present (CP) with half of the cows within steroid group. At 50 h after device removal, 500 micrograms of GnRH was given (iv) to all cows, and weaned calves were reunited with their dams. The induced CL had a normal life span (greater than 16 d) in 17 and 86% (trial 1) and 8 and 79% (trial 2) of NP and P4 cows, respectively. Calf removal did not affect (P greater than .10) the life span of the CL. In Exp. 2, spring-calving multiparous cows (d 18 to 24 pp; d 0) received either no P4 (NP; n = 19), P4 for 6 d via intravaginal devices (P4H; n = 19) or a single im injection of 300 mg P4 (P4 IM; n = 18). At 48 h after device removal or at 8 d after the injection of P4, half of the cows within steroid group received either 500 micrograms GnRH or saline. Corpora lutea had a normal life span in 0, 11, and 80% of NP, P4 IM and P4H cows, respectively, that received GnRH and in 22% of P4-saline cows. In Exp. 3, fall-calving multiparous and primiparous cows (d 25 to 31 pp) received either no progestogen (NP; n = 20), P4 via intravaginal devices for 5 d (P4H; n = 21) or melengestrol acetate (MGA; .5 mg.head-1.d-1 for 5 d orally, n = 15). At 48 d after device removal or at 72 h after the last MGA feeding, all cows received 500 micrograms GnRH. Progesterone post-GnRH injection was increased (greater than 1 ng/ml) at d 7 in 64, 100 and 100%, and remained elevated at d 14 in 11, 46 and 100% of NP, MGA and P4H cows, respectively. For all experiments plasma P4 was increased (range 2 to 5 ng/ml) when the devices containing P4 were in place, then decreased (less than 1 ng/ml) by 48 to 50 h after device removal.(ABSTRACT TRUNCATED AT 400 WORDS)

Endocrine profiles associated with life span of induced corpora lutea in postpartum beef cows.

Two experiments were designed to examine whether hormonal profiles were related to luteal life span in pluriparous postpartum anestrous beef cows. Cows (Exp. 1, n = 34; Exp. 2, n = 23) received norgestomet (N) for 9 d or served as controls (C). Each cow received 1,000 IU human chorionic gonadotropin (hCG) 48 h after removal of N (d 0). Blood samples collected every 15 min for 8 h on d -5, 3 and 5 (Exp. 1) or on d -10 and -1 (Exp. 2) were assayed for luteinizing hormone (LH) and follicle stimulating hormone (FSH). Cortisol was determined in hourly samples collected on d -5 and in samples collected every 2 min during suckling on the same day (Exp. 1). Concentrations of 15-keto-13,14-dihydro-PGF2 alpha (PGFM) were determined in samples collected at 15-min intervals for 2 h on d -5, 3, 5 and 10 (Exp. 1). Estradiol-17 beta was measured in samples collected on d -5 (Exp. 1) or on d -10 and -1 (Exp. 2). Life span of induced corpora lutea was longer (P less than .05) in N than C cows. Percentages of N cows in which corpora lutea, formed in response to hCG, exhibited a normal life span were 83% on farm 1 and 25% on farm 2 (Exp. 1), and 90% (Exp. 2), compared with 0% in C cows. Concentrations of FSH were not affected by N but were lower (P less than .05) on d -5 in cows on farm 2 (.6 +/- .1 ng/ml) than in cows on farm 1 (.8 +/- .1 ng/ml). On d -5, a treatment X farm interaction (P less than .05) for mean LH was observed and frequency of pulses of LH was higher (P less than .01) in N than C cows (2.7 +/- .4 vs. .8 +/- .8 pulses/8 h). Neither cortisol nor PGFM was affected by N. Estradiol was increased in d -1 (6.1 +/- .5 vs 2.6 +/- .8 pg/ml; P less than .01) by N. It is suggested that pre-treatment with N enhanced life span of induced corpora lutea, in part, by influencing secretion of LH and development of follicles, but a threshold concentration of FSH was required for N to exert this effect.

Estrogen and progesterone concentrations in bovine milk during the estrous cycle.

Estrogen and progesterone concentrations in milk during the estrous cycle were estimated in 18 normally cycling Holstein dairy cows. The estrogen and progesterone concentrations in milk during the estrous cycle followed the pattern described for them in blood in the corresponding period. During most of the estrous cycle, estrogen concentration remained at approximately 200 pg/ml and reached a proestrous peak of 360 +/- 127 pg/ml on day 19. The progesterone concentration in milk during the estrous cycle increased to a peak on day 13 (45.5 +/- 6.6 ng/ml) and thereafter declined towards estrus. Estrus detection/prediction based on milk progesterone concentrations appears feasible in view of the significant differences in milk progesterone concentrations between the early luteal (post-ovulatory), luteal and rapid follicular growth periods of the estrous cycle.

Hormonal induction of lactation: estrogen and progesterone in milk.

Estrogen and progesterone in milk during the first 21 days of induced and postpartum lactation in Holstein cows and heifers were estimated by assay procedures. Lactation was induced with estradiol-17 beta and progesterone treatment for 7 days. Estrogen and progesterone in induced lactations differed from concentrations in postpartum lactations. In early lactation estrogen was higher in postpartum milk (521 +/- 103 pg/ml on day 1) than in induced milk (336 +/- 46 pg/ml on day 1), but after day 7 the reverse was true (192 +/- 33 pg/ml and 233 +/- 32 pg/ml on day 7). Progesterone remained higher in induced lactation through the first 21 days than in postpartum lactation with the exception of day 19. Progesterone in postpartum milk increased from 4 +/- 1 ng/ml on day 1 to 11 +/- 2 ng/ml on day 21. Progesterone in induced milk showed greater fluctuation (11 +/- 3 ng/ml on day 1 and 22 +/- 9 ng/ml on day 3) but gradually decreased to 12 +/- 2 ng/ml on day 21 (11 +/- 2 ng/ml on day 21 of postpartum lactation).

Testosterone levels in the blood plasma of male chicken-pheasant hybrids.

Testosterone levels in the blood plasma of sterile male chicken-pheasant hybrids were compared with plasma testosterone levels of roosters and male pheasants, using radioimmunoassay. Mean values of testosterone in the roosters, male pheasants and chicken-pheasant hybrids during spring were 155.47 +/- 4.03, 24.29 +/- 1.55 and 0.68 +/- 0.05 ng./100 ml., respectively and the corresponding values for winter were 88.88 +/- 4.10, 0.61 +/- 0.04 and 0.76 +/- 0.07 ng./100 ml. The pheasant testes were inactive in winter but active in spring whereas the hybrid testes were inactive in winter and spring unlike those of roosters which were active during these seasons. The low levels of testosterone in hybrids during both seasons could be attributed to the impairment of steroid biosynthetic activity of the Leydig cells and may be causally related to the absence of secondary sexual characteristics and the interruption of spermatogenesis in chicken-pheasant hybrids.

Response of bovine serum prolactin and growth hormone to duodenal, abomasal, and oral administration of thyrotropin-releasing hormone.

Thyrotropin-releasing hormone was injected into the duodenum of two 500-kg steers, placed into the abomasum of two prepubertal bulls, and fed to four bull calves (1 to 3 wk of age) to test the effect on concentrations of prolactin and growth hormone in blood serum. Before 20 and 200 mg of thyrotropin-releasing hormone were injected into the duodenum, prolactin in serum averaged 7.5 and 9.4 ng/ml and increased to 52.5 and 129.6 ng/ml at 45 and 35 min after treatment. Average growth hormone concentration of serum was increased also, but the response was more variable than prolactin. Peak concentrations of prolactin and growth hormone in blood serum were 5 to 10 times greater after treatment with thyrotropin-releasing hormone (40 mg/100 kg body weight into abomasum) than before treatment. Within 30 min after oral administration of thyrotropin-releasing hormone (0, .5, 1, and 2 mg/kg body weight) growth hormone concentration of serum was 30, 306, 356, and 317% greater than pretreatment. Prolactin concentration of serum, however, was increased in only one calf.

TRH-stimulation of prolactin release from bovine pituitary cells.

Bovine anterior pituitaries were dispersed with collagenase and the cells grown to monolayers. On day 3 or 4 or culture, cells (4 flasks/treatment group) were incubated for 2 hr with TC medium 199, then with TC medium 199 containing thyrotropin releasing hormone (TRH) for 2 hr. Relative to to values during the first 2-hr incubation, prolactin concentration in media from cow pituitary cell cultures exposed to 0 (control), 0.01, 0.1 or 10.0 ng TRH/ml medium average -23, 799, 1966, 1926 and 1976 ng/ml. Relative to control, each dose of TRH increased (P less than 0.01) media prolactin concentration. Comparable values for growth hormones were -5, 9, 12, 21, and 21 ng/ml and the increase in growth hormone release relative to controls was significant (P less than 0.05). A second experiment, designed to determine whether TRH would increase prolactin release by pituitary cells from cows, steers and a bull was conducted using the same procedures. Changes in media prolactin concentration (ng/ml) after 0, 0.01, 0.1, 1.0, 10.0 and 100.0 ng TRH/ml medium was -27, 63, 109, 207, 226 and 137 for cows and -14, 13, 36, 153, 166 and 80 for steers. After 0.0, 10 and 100 ng TRH/ml medium comparable values for a bull were -6, 14 and 31 ng/ml. Growth hormone (ng/ml) in these media was not different from controls. We conclude that TRH stimulates prolactin release from pituitary cells of cows, steers and bulls but its effect on growth hormone release is not consistent.