Effects of pituitary adenylate cyclase-activating polypeptide (PACAP), prostaglandin E2 (PGE2) and growth hormone releasing factor (GRF) on the release of growth hormone from cultured bovine anterior pituitary cells in vitro

The effect of pituitary adenylate cyclase-activating polypeptide (PACAP) on growth hormone (GH) release was compared with that of prostaglandin E₂ (PGE₂) and growth hormone releasing factor (GRF) from cultured bovine anterior pituitary cells in vitro. Both PACAP and PGE₂ stimulated GH release at concentrations as low as 10⁻⁹ and 10⁻⁸ M, respectively, (P<0.01). However, GRF released GH at a concentration as low as 10⁻¹³ M (P<0.01). Percent increases of GH compared with controls were not significantly different among GRF, PACAP, and PGE₂ at 10⁻⁷ M; however, the increases of GH by the 10⁻⁸ M GRF, PACAP and PGE₂ were 196, 118, and 27%, respectively, (P<0.01), and 124, 65, and 1% in the 10⁻⁹ M media, respectively, (P<0.01). When GRF and somatostatin (SS) were added together, the GH releasing effect of GRF was blunted (P<0.01). Similar bluntness were observed in PACAP and PGE₂, when SS was added. The stimulatory effects of GRF and PGE₂ together were similar to that by either GRF or PGE₂ alone. When GRF and PACAP were added together, the GH released by both secretagogues was greater than that by PACAP alone (P<0.01); however, a synergistic effect was not clear when compared with GRF alone.

These findings suggest that PACAP and PGE₂ may modulate the release of GH in cattle.     

Modulation of growth hormone-releasing factor-induced release of growth hormone from bovine pituitary cells

Growth hormone (GH)-releasing factor (GRF) at concentrations of 10−¹² through 10−⁷M for 6 hr linearly increased GH release (b₁ = 10.4 ± .3) from bovine anterior pituitary cells in culture. Maximum release of GH (262% above controls) occurred at 10−⁷M GRF. In contrast, GH release-inhibiting factor (SRIF) at 10−¹² through 10−⁵M had no effect on basal concentrations of GH. In a second experiment, as the proportion of SRIF relative to GRF increased. SRIF suppression of GRF-induced GH release from anterior pituitary cells increased. In a third experiment, anterior pituitary cells cultured in media containing fetal calf serum (FCS) were treated with cortisol (0 or 10 ng/ml media) for 24 hr before exposure to 10−¹³ through 10−⁷M GRF. GRF linearly increased GH secretion (b₁ = 7.4 ± .3) and cortisol augmented this response (b₁ = 10.5 ± .6). However, when cells were cultured in media containing dextran-charcoal treated FCS, cortisol did not alter GRF-induced GH release. Our results demonstrate that GH response of bovine anterior pituitary cells to GRF was modulated negatively by SRIF. However, augmentation of GRF-induced GH release by cortisol was evident only when cells were cultured in media supplemented with untreated FCS.     

Dose effect of human growth hormone-releasing factor and thyrotropin-releasing factor on hormone concentrations in lactating dairy cows

Two experiments were conducted to study the effects of growth hormone-releasing factor (GRF) and thyrotropin-releasing factor (TRF) administration on hormone concentrations in dairy cows. In the first trial, 12 cows were used on 5 consecutive days to determine the effect of four sc doses of GRF (0, 1.1, 3.3 and 10 μg•kg⁻¹ BW) and three sc doses of TRF (0, 1.1 and 3.3 μg•kg⁻¹ BW) combined in a factorial arrangement. GRF and TRF acted in synergy (P = .02) on serum growth hormone (GH) concentration even at the lowest dose tested and GH response to the two releasing factors was higher than the maximal response observed with each factor alone. TRF increased (P<.01) prolactin (Prl), thyrotropin (TSH), triiodothyronine (T₃) and thyroxine (T₄) concentrations similarly at the 1.1 and 3.3 μg•kg⁻¹ doses and GRF did not interact (P>.40) with TRF on the release of these hormones. In the second trial, the effect of GRF (3.3 μg•kg⁻¹ BW, sc) and TRF (1.1 μg•kg⁻¹ BW, sc) was tested at three stages (18, 72 and 210 days) of lactation on serum Prl and TSH concentrations. Eighteen cows (n = 6 per stage of lactation) were used in two replicates of a 3 × 3 latin square. The TRF and GRF-TRF treatments were equipotent (P>.05) in increasing Prl and TSH concentrations. Prl and TSH responses were similar (P>.40) throughout lactation. In summary, GRF at doses ranging from 1.1 to 10.0 μg•kg⁻¹ and TRF at doses ranging from 1.1 to 3.3 μg•kg⁻¹ act in synergy on GH release and do not interact on Prl, TSH, T₃ and T₄ concentrations in dairy cows. Furthermore, Prl and TSH response to TRF are not affected by stage of lactation.     

Effect of a nonsteroidal aromatase inhibitor on in vitro and in vivo secretion of estradiol and on the estrous cycle in ewes.

Three experiments were performed to study effects of decreased concentrations of estradiol-17β (E₂) on lifespan and function of ensuing ovine corpora lutea (CL). In experiment 1, 52 follicles were collected from 10 ewes and placed into individual culture with 0 or .01 μCi ³H-androstenedione (10 ng; ³H-A) and 0, 10⁻¹¹, 10⁻⁹, 10⁻⁷, or 10⁻⁵ M of a nonsteroidal aromatase inhibitor, CGS16949A (CGS). Concentrations of E₂ secreted into the medium, and synthesis of estrogens as estimated by formation of ³H-water from ³H-A were decreased by 10⁻⁵ and 10⁻⁷ (P<.01), but not 10⁻⁹ or 10⁻¹¹ M CGS. In experiment 2, luteolysis was induced in 24 ewes by injection of PGF₂ on days 5 to 10 of the estrous cycle (0 hr). Ewes received 0, 0.5, 1.0, 2.0 or 4.0 mg CGS per kg BW i.v. at −12, 0, 12 and 24 hr, and an ovulatory dose of hCG at 36 hr. Jugular (P<.001) and vena caval (P<.001) concentrations of E₂ were decreased by CGS at all doses tested for 8 to 10 hr, but had returned to levels similar to control ewes by the time of the next injection. Concentrations of E₂ around the time of the LH surge were similar in control and treated ewes. During the subsequent luteal phase, concentrations of progesterone (P₄) were similar in control and treated ewes. Thus, transient decreases in E₂ during the follicular phase were not deleterious to the subsequent luteal phase. In experiment 3, luteolysis was induced in 18 ewes by injection of PGF₂ on days 6 or 7 (0 hr) of the estrous cycle. Ewes received 0 or 1 mg CGS per kg BW i.v. every 8 hr from 0 to 40 hr. Ovulation was induced with hCG at 36 hr. CGS reduced jugular (P<.001) and vena caval (P<.001) concentrations of E₂, prevented an endogenous surge of LH (P<.05) and increased (P<.001) concentrations of FSH. All ewes had ovulated a marked follicle by 72 hr, but onset of the luteal phase, as assessed by concentrations of P₄, was delayed (P<.01) in ewes receiving CGS. Delayed luteal phases were not solely attributable to the presence of new CL or to luteinization of follicular cysts. When data were aligned according to the day ewes were observed in estrus, profiles of P₄ did not differ with treatment. Therefore, normal luteal function ensued following estrus whether or not ewes re-ovulated. In conclusion, decreased secretion of E₂ by the preovulatory follicle was not involved in the ontogeny of CL of short lifespan or subnormal function. Instead, adequate production of E₂ or precisely timed E₂ secretion may be required during follicular development for subsequent functional luteinization.     

Opioid modulation of gonadotropin releasing hormone release from the hypothalamic preoptic area in the pig

Two experiments (Exp) were conducted to examine in vitro the release of gonadotropin releasing hormone (GnRH) from the hypothalamus after treatment with naloxone (NAL) or morphine (MOR). In Exp 1, hypothalamic-preoptic area (HYP-POA) collected from 3 market weight gilts at sacrifice and sagitally halved were perifused for 90 min prior to a 10 min pulse of morphine (MOR; 4.5 × 10⁻⁶ M) followed by NAL (3.1 × 10⁻⁵ M) during the last 5 min of MOR (MOR + NAL; N=3). The other half of the explants (n=3) were exposed to NAL for 5 min. Fragments were exposed to KCl (60 mM) at 175 min to assess residual GnRH releasability. In Exp 2, nine gilts were ovariectomized and received either oil vehicle im (V; n=3); 10 μg estradiol-17β/kg BW im 42 hr before sacrifice (E; n=3); .85 mg progesterone/kg BW im twice daily for 6 d prior to sacrifice (P₄; n=3). Blood was collected to assess pituitary sensitivity to GnRH (.2 μg/kg BW) on the day prior to sacrifice. On the day of sacrifice HYP-POA explants were collected and treated as described in Exp 1 except tissue received only NAL. In Exp 1, NAL increased (P<.05) GnRH release. This response to NAL was attenuated (P<.05) by coadministration of MOR. Cumulative GnRH release after NAL was greater (P<.05) than after MOR + NAL. All tissues responded similarly to KCl with an increase (P<.05) in GnRH release. In Exp 2, pretreatment luteinizing hormone (LH) concentrations were lower (P<.05) in E gilts compared to V and P₄ animals with P₄ being lower (P<.05) than V gilts. LH response to GnRH was lower (P<.05) in E pigs than in V and P₄ animals, while the responses was similar between V and P₄ gilts. NAL increased GnRH release in all explants, whereas, KCl increased GnRH release in 6 of 9 explants. These results indicate that endogenous opioid peptides may modulate in vitro GnRH release from the hypothalamus in the gilt.     

Effect of daily injections of growth hormone-releasing factor and thyrotropin-releasing hormone on growth and endocrine parameters in chickens

The control of growth is a complex mechanism regulated by several metabolic hormones including growth hormone (GH) and thyroid hormones. In avian species, as well as in mammals, GH secretion is regulated by hypothalamic hypophysiotropic hormones. Since thyrotropin-releasing hormone (TRH) and growth hormone-releasing factor (GRF) are potent GH secretagogues in poultry, we were interested in determining the influence of daily intravenous administration of either peptide or both simultaneously on circulating GH and IGF-I concentrations and whether an improvement in growth rate or efficiency would be obtained.

Male broiler chicks were injected once daily for a period of 21 days with either GRF (10 μg/kg), TRH (1 μg/kg) or both GRF and TRH (10 and 1 μg/kg respectively) between four and seven weeks of age. On the last day of the experiment, following intravenous injection of TRH, GRF or a combination of GRF and TRH, plasma GH levels were significantly (P<.05) increased to a similar extent in control chicks and in those which had received daily peptide injections for the previous 21 days. Circulating GH levels between 10 and 90 min post-injection were significantly (P<.05) greater and more than additive than GH levels in chicks injected with both GRF and TRH when compared to those injected with either peptide alone. Mean plasma T₃ concentrations during that same time period were significantly elevated (P<.05) above saline-injected control chick levels in birds treated with TRH or GRF and TRH respectively, regardless of whether the chicks had received peptide injections for the previous 21 days. There was no evidence of pituitary refractoriness to chronic administration of either TRH or GRF injection in terms of growth or thyroid hormone secretion.

Despite the large elevation in GH concentration each day, growth rate, feed efficiency and circulating IGF-I concentrations were not enhanced. Thus the quantity or secretory pattern of GH secretion induced by TRH or GRF administration was not sufficient to increase plasma IGF-I concentration or growth.     

Influence of age and sex on basal secretion of growth hormone (GH) and on GH-induced release by porcine GH-releasing factor pGRF(1–29NH2) in growing pigs

The aim of this study was to determine the effect of age and sex on basal secretory patterns of growth hormone (GH) and growth hormone-releasing factor (GRF) induced GH release. Eighteen pigs (9 castrated males and 9 females) were stimulated with pGRF(1–29)NH₂ at 7,11,15,19 and 23 weeks of age. Blood samples were taken from each animal via jugular vein cannulate every 20 min, from 6 hr before to 5 hr after iv GRF administration at a dose of 4 μg/kg. GH baseline levels, amplitude of the GH peaks, area under the GH peaks and the overall mean of GH serum levels decreased (P<.001) with age in both sexes. Age also had a marked effect on GRF-induced GH release: the amplitude of GH peaks and area under the GH peaks decreased (P<.001) with age. The GH response to pGRF(1–29)NH₂ varied considerably, depending on the timing of the episodic endogenous secretion of GH. An immediate response (<30 min) was observed when GRF was injected at the end of a trough period or at the beginning of a peak, but there was no immediate response when GRF was injected at the end of a peak or at the beginning of a trough period. Our results show that both endogenous GH secretion and pGRF(1–29)NH₂-induced GH release declines with age, suggesting a decreased sensitivity of the somatotroph cells to GRF with age; and that the high variability of the GH response to pGRF(1–29)NH₂ stimulation depends greatly on the timing of the episodic endogenous GH release, thus implying a possible episodic endogenous somatostatin secretion by the hypothalamus.     

Hormonal changes following an acute stress in control and somatostatin-immunized pigs

Sixteen Yorkshire pigs (49 ± 2 kg BW at 17 weeks) were immunized against somatostatin (SRIF; 4 males, 4 females) or its conjugated protein, bovine serum albumin (BSA; controls; 4 males, 4 females). Immunizations were done at 10, 12 and 14 weeks of age. Jugular vein cannulae were surgically inserted at 17 weeks of age. Five d later, half of each sex from the control and SRIF-immunized groups were stressed. The other half were subjected to the same stress 48 hr later. On both days, remaining animals were used as unstressed controls. The stress consisted of 5 min of snare restraint. Blood samples were collected from all pigs on both days at −20, −15, −10, −5, 0 (beginning of stress), 2, 6, 10, 15, 20, 30, 40, 60, 90, 120, 150, 180 and 240 min. Samples were radioimmunoassayed for cortisol, growth hormone (GH), prolactin (Prl), insulin, triiodothyronine (T₃), thyroxine (T₄) and insulin-like growth factor I (IGF-I). Mean antibody titers against SRIF (1:150 dilution) at 15 weeks were 0.49 ± .09% and 54.5 ± 4.9% for control and SRIF immunized pigs, respectively. Gender and immunization against SRIF had no effect on any of the variables measured (P>0.05), except for T₃ levels which were greater in females than in males (P<0.05). The stress by time of sampling interaction was significant (P<0.01) for all hormones measured. Cortisol values almost tripled within 15 min of stress, reaching concentrations above 100 ng/mL. Maximal increases were seen at 2 min for T₄ (14%), at 6 min for T₃ (36%), at 15 min for Prl (46%) and at 10 min for insulin (141%). An increase of 129% in GH concentration was present at 20 min in stressed pigs; however, an increase of 97% was also seen at 120 min in control pigs. Concentrations of IGF-I decreased (21%) by 60 min in the stressed pigs and remained depressed for up to 150 min. Stress associated with snare restraint, therefore, induces major changes in the concentrations of a series of hormones in growing pigs. On the other hand, immunization against SRIF did not alter any of the hormonal profiles measured. Since snare restraint is widely used to handle pigs during jugular puncture, any study of hormonal secretion in this species should be carried out under carefully controlled conditions in terms of blood sampling technique.     

Relationships between the thyroid and somatotropic axes in steers I: Effects of propylthiouracil-induced hypothyroidism on growth hormone, thyroid stimulating hormone and insulin-like growth factor I

The effects of propylthiouracil (PTU)-induced thyroid hormone imbalance on GH, TSH and IGF-I status in cattle were examined. In the first study, four crossbred steers (avg wt 350 kg) were fed a diet dressed with PTU (0, 1, 2 or 4 mg/kg/d BW) in a Latin square design with four 35-d periods. On day 29 in each period, steers were challenged with an intrajugular bolus of thyrotropin releasing hormone (TRH, 1.0 μg/kg). Blood samples were obtained to assess the change in plasma GH and TSH as affected by PTU. Plasma IGF-I was measured from blood samples obtained before and after (every 6 hr for 24 hr) intramuscular injection of bovine GH (0.1 mg/kg, day 31). Doses of 1 and 2 mg/kg PTU increased plasma T₄ (P<.01). At 4 mg/kg, PTU depressed T₄ concentrations to 30% of control (P<.01). Plasma T₃ linearly decreased with increasing doses of PTU (P<.01). Plasma TSH increased when PTU was fed at 4 mg/kg (P<.05) while the TSH response to TRH declined with increasing PTU (P<.02). Neither basal nor TRH-stimulated plasma concentration of GH was affected by PTU; the IGF-I response to GH tended to increase at the 1 and 2 mg/kg PTU (P<.01). In a second study 24 crossbred steers were fed PTU (1.5 mg/kg) for 119 d in a 2 × 2 factorial design with implantation of the steroid growth effector, Synovex-S (200 mg progesterone + 20 mg estradiol), as the other main effect. Basal plasma GH and IGF-I were not affected by PTU treatment. Synovex increased plasma concentration (P<.01) of IGF-I without an effect on plasma GH. The data suggest that mild changes in thyroid status associated with PTU affects regulation of T₃, T₄ and TSH more than GH or IGF-I in steers.     

Effects of chronic GRF treatment on lambs having low or normal birth weight

The effects of a long term treatment with human GRF(1–29)NH2 on plasma growth hormone (GH), somatomedin C (Sm-C), histomorphometric parameters of bone growth and body composition were investigated in normal and low birthweight male lambs. The animals were divided into two groups according to their birthweight: 24 normal birthweight (NBW) lambs weighing more than 4 kg and 22 low birthweight (LBW) lambs weighing less than 2.5 kg at birth. Half of the animals in each group received two daily subcutaneous injections (8 μg/kg body weight) of hGRF(1–29)NH2 (GRF) from birth to slaughter at 45 or 90 days of age. The other animals received the solvant only. At the beginning and at the end of the treatment, plasma GH and serum Sm-C concentrations were measured in all groups. After slaughter, a histomorphometric study was performed on undecalcified sections of metacarpal growth plates, and the remaining of the carcass was pulverized to study the chemical body composition.

GRF induced GH release in both GRF-treated groups. However, plasma GH reached higher (P<.001) concentrations and the GRF-induced GH peak lasted longer in LBW than in NBW lambs. At day 45, the GRF treatment increased (P<.05) serum Sm-C concentrations in LBW. Most of histomorphometric parameters reflecting the metacarpal growth in length, were not statistically modified under GRF treatment. However, the size of degenerative cells was smaller (P<.05) in LBW treated lambs as compared to controls. Consequently, the cell production in the growth plate was increased (P<.05) under GRF treatment. In both NBW and LBW groups at 45 days of treatment, GRF treatment reduced the amounts of lipids (P<.025) and energy (P<.05), while increased (P<.01) phosphorus deposition in the body. In contrast, there was no effect of GRF treatment on protein content.

We conclude from this experiment that the induction of GH secretion by a chronic treatment with GRF is able to modify some patterns of growth. However, most of the effects of GRF were observed in LBW lambs and after 45 days of treatment only. This suggests that treatment with GRF may serve to compensate for growth in growth-retarded animals. Further studies with different mode of GRF administration should indicate whether it is as much effective in normal animals.     

Perturbed metabolism and hormonal profiles in calves infected with Sarcocystis cruzi

Plasma concentrations of growth hormone (GH), thyroid stimulating hormone (TSH), insulin (IN), thyroxine (T₄), and triiodothyronine (T₃) in addition to metabolic parameters [N balance (NB), urinary 3-methylhistidine (TMH), urinary creatinine (CR), and urinary hydroxyproline (HP)] were measured in 4-mo-old Holstein steers divided equally among groups infected with Sarcocystis (I), noninfected ad libitum fed (C), and noninfected pair fed to I (PF) (7 steers per treatment). Effects of infection on these parameters beyond those attributable to altered dietary intake were determined using orthogonal contrasts (effect of intake, C vs I + PF; effect of infection, PF vs I). NB was higher in C than I and PF (P<.05) and lower in I than PF (P<.02). Hydroxyproline and CR were influenced by intake (P<.05) and HP excretion was reduced in association with infection (P<.05). Reduced intake was associated with lowered mean basal plasma concentrations of GH, IN, T₃ and T₄ (P<.05). Infection further reduced (P<.001) plasma T₃ concentration.

Triiodothyronine and T₄ responses following an intravenous bolus of thyrotropin releasing hormone (TRH) were measured. The magnitude of the responses in I and PF were lower than those observed in C (P<.05). Plasma T₃ responses were further reduced in association with infection (P<.05). Insulin responses to intravenous arginine infusion (ARG) were also low in association with reduced intake. Growth hormone responses to TRH or ARG were affected by the level of feed intake only. These data suggest that hormonal perturbations associated with the insult of infection further compromise metabolism and the direction of nutrient partitioning that would ordinarily be associated with developmental growth in young steers beyond those responses anticipated from solely the reduction of feed intake.     

Macrolide-lincosamide resistance determinants in streptococcal species isolated from the bovine mammary gland

Seventy one streptococci isolated from dairy cows with clinical mastitis were tested for erythromycin and lincomycin susceptibility. Ten isolates (7.1%) were resistant to erythromycin and/or lincomycin and seven were constitutive and three were inducibly resistant. Nine of the isolates hybridized with one or more of the Erm probes tested and eight isolates gave PCR products with rRNA methylase primers. The Erm determinants were transferable at frequency of 10⁻⁵ to 10⁻⁶ per recipient.     

Stimulating effects of androgen on proliferation of cultured ovarian germ cells through androgenic and estrogenic actions in embryonic chickens

The effect of androgen on germ cell proliferation was evaluated by a chicken ovarian germ–somatic cell co-culture model and the mechanisms were explored. Ovarian cells were dispersed from 18-day-old embryos, cultured in serum-free McCoy's 5A medium and challenged with testosterone (T) alone or in combinations with androgen receptor antagonist Flutamide, estrogen receptor antagonist Tamoxifen or aromatase inhibitor Letrozole for 48 h. Germ cells were identified by c-kit immunocytochemistry. The number of germ cells was counted and the proliferating cells were identified by immunocytochemistry of proliferating cell nuclear antigen (PCNA). The labeling index (LI) was determined for germ cells. Results showed that T (10⁻⁷ to 10⁻⁶ M) significantly increased the number of germ cells (P < 0.05) and this stimulating effect was inhibited by Flutamide (10–1000 ng/ml), Tamoxifen (10–1000 ng/ml) or Letrozole (10⁻⁹ to 10⁻⁷ M) in a dose-dependent manner. Furthermore, PCNA-LI of germ cells displayed similar changes with the numbers of germ cells. These results indicated that T-stimulated proliferation of cultured ovarian germ cells through both, androgenic and estrogenic actions in embryonic chickens.     

Extension of short cycles in postpartum beef cows by intrauterine treatment with catecholestradiol

Eight multiparous beef cows were used to examine the effects of intrauterine infusion of catecholestradiol (4-hydroxylated estradiol) on development and function of the first corpus luteum after parturition. Calves were weaned on day 1 (day 0 = parturition) to initiate formation of a corpus luteum (CL) by approximately day 10 or 11. Before CL formation, on days 5 to 9, cows received twice daily infusions of catecholestradiol (4 μg; n = 4) or vehicle (n = 4) into the uterine horn opposite the previous pregnancy. Plasma progesterone during the first estrous cycle was elevated longer (P<.001) and reached a higher (P<.001) concentration in cows treated with catecholestradiol. The decline in progesterone was associated with an increase in plasma 13,14-dihydro, 15-keto-prostaglandin F₂ (PGFM) in all cows infused with catecholestradiol. In contrast, a rise in PGFM at the end of the first short cycle was detected in only one of four cows treated with vehicle. Furthermore, PGFM concentrations were linearly related (R² = .870; P<.001) to concentrations of progesterone. Estradiol-17β concentrations were not different during the infusion period, but after formation of the first CL, estradiol remained elevated (P<.01) in cows that received vehicle. Results of this experiment suggest that exposure of postpartum beef cows to catecholestradiol extended luteal function in association with enhanced PGFM release.     

Effects of cimaterol administration on plasma concentrations of various hormones and metabolites in friesian steers

The aim of the experiment was to determine the acute and chronic effects of the β-agonist, cimaterol, on plasma hormone and metabolite concentrations in steers. Twelve Friesian steers (liveweight = 488 ± 3 kg) were randomly assigned to receive either 0 (control; n=6) or .09 mg cimaterol/kg body weight/day (treated; n=6). Steers were fed grass silage ad libitum. Cimaterol, dissolved in 140 ml of acidified distilled water (pH 4.2), was administered orally at 1400 hr each d. After 13 d of treatment with cimaterol or vehicle (days 1 to 13), all animals were treated with vehicle for a further 7 d (days 14 to 20). On days 1, 13 and 20, blood samples were collected at 20 min-intervals for 4 hr before and 8 hr after cimaterol or vehicle dosing. All samples were assayed for growth hormone (GH) and insulin, while samples taken at −4, −2, 0, +2, +4, +6 and +8 hr relative to dosing were assayed for thyroxine (T₄), triiodothyronine (T₃), cortisol, urea, glucose and non-esterified fatty acids (NEFA). Samples taken at −3 and +3 hr relative to dosing were assayed for IGF-I only. On day 1, cimaterol acutely reduced (P<.05) GH and urea concentrations (7.6 vs 2.9 ± 1.4 ng/ml; and 6.0 vs 4.9 ± 0.45 mmol/l, respectively; mean control vs mean treated ± pooled standard error of difference), and increased (P<.05) NEFA, glucose and insulin concentrations (160 vs 276 ± 22 μmol/l, 4.1 vs 6.2 ± 0.15 mmol/l and 29.9 vs 179.7 ± 13.9 μU/ml, respectively). Plasma IGF-I, T₃, T₄ and cortisol concentrations were not altered by treatment. On day 13, cimaterol increased (P<.05) GH and NEFA concentrations (7.7 vs 14.5 ± 1.4 ng/ml and 202 vs 310 ± 22 mEq/l, respectively) and reduced (P<.05) plasma IGF-I concentrations (1296 vs 776 ± 227 ng/ml). Seven-d withdrawal of cimaterol (day 20) returned hormone and metabolite concentrations to control values. It is concluded that : 1) cimaterol acutely increased insulin, glucose and NEFA and decreased GH and urea concentrations, 2) cimaterol chronically increased GH and NEFA and decreased IGF-I concentrations, and 3) there was no residual effect of cimaterol following a 7-d withdrawal period.     

Effect of exogenous thyrotropin-releasing hormone (TRH) administration on plasma levels of triiodothyronine (T3), thyroxine (T4) and growth hormone (GH) in chronically catheterized suckling piglets.

The purpose of the present study was to determine experimental conditions to stimulate secretion of thyroid hormones (T₃ and T₄) with thyrotropin-releasing hormone (TRH) injections in suckling piglets during the first weeks of postnatal life. Three consecutive experiments were conducted. Four 10–20 d old piglets were i.m. injected with 0, 20, 100, 500 μg (experiment 1) or 0, 4, 20, 100 μg TRH/kg BW (experiment 2) according to a 4 × 4 latin square design involving different litters in each experiment. Blood samples were taken −15, −1, 15, 30, 45, 60, 90, 120 180 and 300 min after TRH injection in experiment 1, and −.25, −.08, .25, .5, 1, 2, 4, 6, 8, 12, 24, 30, 36, 48, 60 and 72 hr after TRH injection in experiment 2. T₃ and T₄ levels were significantly (P<.01) increased as soon as 30 and 45 min after TRH injection, respectively. Maximal levels of T₃ and T₄ were obtained 2 and 4 hr after the injection of 100 μg TRH. T₃ and T₄ returned to basal levels within 6 and 8 hr post injection, respectively. Plasma pGH levels were significantly (P<.001) increased 15 min after TRH injection in piglets injected with 500 μg. In experiment 3, 100 μg TRH/kg BW were injected i.m. either daily or every other day from .0 to 23 days of age. Results showed that T₄ response to TRH did not decrease after repeated injections. These results indicate that daily i.m. injections of 100 μg TRH/kg BW can be used to increase thyroid hormone levels for at least 13 d in the young suckling piglet.     

Perforin expression can define CD8 positive lymphocyte subsets in pigs allowing phenotypic and functional analysis of natural killer, cytotoxic T, natural killer T and MHC un-restricted cytotoxic T-cells

In this study we have used the expression of perforin to characterize subsets of porcine cytotoxic lymphocytes. Perforin positive lymphocytes expressed both CD2 and CD8, most were small dense lymphocytes (SDL) and up to 90% were CD3 negative. However, the numbers of perforin positive T-cells increased with the age of the animal and their populations increased after specific antigen stimulation in vitro. The remaining perforin positive lymphocytes were large and granular and contained more CD3⁺CD5⁺CD6⁺ T-cells (−40%) of which a substantial proportion also co-expressed CD4. Perforin was expressed in subpopulations of both CD8 and CD8β lymphocytes, but was not expressed in γδ T-cells or monocyte/macrophages. The perforin positive CD3⁻ subset was phenotypically homogeneous and defined as CD5⁻CD6⁻CD8β⁻CD16⁺CD11b⁺. This population had NK activity and expressed mRNA for the NK receptor NKG2D, and adaptors DAP10 and DAP12. Perforin positive T-cells (CD3⁺) could be divided into at least three subsets. The first subset was CD4⁻CD5⁺CD6⁺CD11b⁻CD16⁻ most were small dense lymphocytes with cytotoxic T-cell activity but not all expressed CD8β. The second subset was mainly observed in the large granular lymphocytes. Their phenotype was CD4⁺CD5⁺CD6⁺CD8β⁺CD16⁻CD11b⁻ and also showed functional CTL activity. Thus not all of double positive T-cells are memory helper T-cells. The third subset did not express the T-cell co-receptor CD6, but up to half of them expressed another T-cell co-receptor CD5. The majority of this subset expressed CD11b and CD16, thus the third perforin positive T-cell subset was CD3⁺CD4⁻CD5⁺CD6⁻CD8β^±CD11b⁺CD16⁺, and possessed MHC-unrestricted cytotoxicity and LAK activity.     

Effects of a second mutant allele (V199I) at the PRKAG3 (RN) locus on carcass composition in pigs

The effect of a second mutant allele (V199I, here denoted rn*) at the PRKAG3 (RN) locus on carcass composition was determined in 334 pigs, entire males and females, from crosses between Swedish Hampshire (H) and Finnish Landrace (L) (H × LH; LH × H; LH × LH). Pigs were classified according to DNA test into the following PRKAG3 genotypes: RN⁻/RN⁻ (23%), RN⁻/rn⁺ (24%), RN⁻/rn* (33%), rn⁺/rn⁺ (8%), rn⁺/rn* (9%) and rn*/rn* (2%). The pigs were slaughtered at a commercial slaughterhouse and assessed 24 h postmortem. Right sides were fabricated into primary wholesale cuts, then further processed into defatted hams and loins, and a subset of hams (n = 122) was dissected into the five major individual muscles. The genotype frequencies for the subsample were RN⁻/RN⁻ (27%), RN⁻/rn⁺ (20%), RN⁻/rn* (35%), rn⁺/rn⁺ (9%), rn⁺/rn* (8%) and rn*/rn* (1%). Weights were recorded for meat and bone in ham and loin, fat in ham, back and shoulder and the individual dissected muscles. The genotype effect was significant (P < 0.05) for estimated lean meat content and the proportions of meat and bone and fat in ham and loin (of carcass weight). Also, the content of meat and bone in ham and loin, in proportion of whole ham and loin, respectively, differed significantly (P < 0.01) between genotypes. Estimated lean meat content was highest for RN⁻/RN⁻ (63.0%) and RN⁻/rn⁺ (63.1%) and lowest in the combined group rn*/⁻ (rn⁺/rn* and rn*/rn*, 61.7%); RN⁻/rn* (62.5%) and rn⁺/rn⁺(62.1%) were intermediate. The same results were found for meat and bone in ham and loin, as a proportion of whole ham and loin, respectively. RN⁻/RN⁻ and RN⁻/rn⁺ did not differ in any trait; however, they produced carcasses with the lowest proportions of fat within loin and the major wholesale cuts (ham, loin and shoulder). The carcass percentage of meat and bone in ham was higher in the three RN⁻/ genotypes (RN⁻/RN⁻, RN⁻/rn⁺ and RN⁻/rn*, P < 0.05) than in the rn*/⁻ group, whereas rn⁺/rn⁺ did not (P > 0.05) differ from any of the other genotypes. RN⁻/rn⁺ and RN⁻/rn* had higher (P < 0.05) proportion of meat and bone in loin compared to the rn*/⁻ group. We conclude that the second mutant allele found at the PRKAG3 (RN) locus, rn*, decreased the lean meat content compared with the two other alleles (RN⁻, rn⁺). The RN⁻/RN⁻ and RN⁻/rn⁺ genotypes were leanest, followed by RN⁻/rn* and rn⁺/rn⁺, and rn⁺/rn* and rn*/rn* were the fattest.     

Effets d'une dose unique de dexamethasone sur la production d'anion superoxyde et sur le chimiotactisme des polynucleaires neutrophiles sanguins du chevalEffects of a single injection of dexamethasone on blood neutrophils functions in the horse

Chemotaxis and superoxide anion production of neutrophils in healthy horses were investigated before and 8h after, a single injection of dexamethasone at a dose of 0.045mg/kg. Chemotaxis was studied by the technique of migration under agarose and superoxyde production was measured by ferricytochrome c reduction.

Superoxide anion production was not changed, but the chemotactic index, with zymosan activated horse serum, was increased. The FMLP chemoattractant effect, at 10⁻⁴M, was slightly enhanced.     

Response of young broiler chickens to chronic injection of recombinant-derived human insulin-like growth factor-I

The purpose of this study was to determine if exogenous insulin-like growth factor-I (IGF-I) would improve growth rate or body composition of young broiler chickens. Broiler cockerels were given a daily intramuscular (im) injection of sodium acetate buffer (buffer control), 100 or 200 μg recombinant-derived human IGF-I (rhIGF-I) per kg body weight from 11 to 24 days of age. Exogenous IGF-I did not affect the average daily gain, average daily feed consumption, or the gain-to-feed ratio of broiler chickens. Although daily injection of 200 μg/kg of rhIGF-I reduced (P<0.05) body ash content, there was no significant effect of IGF-I treatment on either body fat or protein content. Plasma GH levels were depressed (P<0.05) by chronic treatment with rhIGF-I. In contrast, plasma levels of T₃ and T₄ were not affected by rhIGF-I treatment. The half-life of rhIGF-I in plasma was determined at 25 days of age in naive control or chronically-injected chickens after a single intravenous dose of 50 μg rhIGF-I/kg. We found a single compartment, first-order disappearance pattern of rhIGF-I from chicken plasma. The half-life (t_1/2) of rhIGF-I in plasma was similar (t_1/2 = 32.5 min) for naive controls (injected once) or chronically-treated chickens which had received a daily injection of rhIGF-I (100 or 200 μg/kg) for 14 d. These data indicate that daily injection of IGF-I cannot be used to enhance growth performance or body composition of broiler chickens when given during the early growth period. The depression of plasma GH levels in rhIGF-I-injected chickens supports a negative-feedback role of IGF-I on pituitary GH secretion.