Calcitonin receptor binding in the hen anterior pituitary during an oviposition cycle

The equilibrium dissociation constant (K_d) and the maximum binding capacity (B_max) of calcitonin (CT) receptor in the plasma membrane of the anterior pituitary in hens were examined by Scatchard analysis of specific binding of ¹²⁵I‐labeled chicken CT. Values of K_d and B_max of CT receptor were smaller in laying hens than in non‐laying hens. A decrease in the K_d and B_max value of CT receptor was observed in the anterior pituitary after the injection of estradiol‐17β and progesterone into nonlaying hens, but not changed after the injection of 5α‐dihydrotestosterone. During an oviposition cycle, the K_d and the B_max value decreased 3 h before oviposition. In non‐laying hens, neither the K_d nor the B_max value changed during a full day period. The present study suggests that the CT action on the anterior pituitary may increase 3 h before oviposition by the effect of estradiol‐17β and progesterone in laying hens.     

Interspecies variations of corticosteroid-binding globulin parameters

In mammalian plasma, cortisol binds to a specific α₁-glycoprotein: corticosteroid-binding globulin (CBG). In this study, we measured the protein binding of cortisol by equilibrium dialysis in seven species in which plasma cortisol concentrations varied from 0.02 to 0.05 (ewe, dog, cow) to 0.1 to 0.6 (horse, human, cynomolgus monkey) to reach 1.6 μM (squirrel monkey). No binding of cortisol to CBG was discernible in plasma from squirrel monkey. In all other species examined, we showed that the CBG maximal capacity (B_max) was 3 (1.7 to 5.2) times more than the plasma cortisol levels, with cow, dog, ewe exhibiting the lowest and cynomolgus monkey exhibiting the highest values. We also noted the existence of a linear relationship between B_max and the corresponding dissociation constant (K_d), B_max being systematically 10 (8.5 to 11.8) times more than K_d. The low binding affinity of cortisol assigned to albumin did not differ between species. The free (6 to 14%), CBG-bound (67 to 87%), and albumin-bound (7 to 19%) cortisol fractions calculated from the estimated binding parameters and measured plasma cortisol concentrations were similar within species, except for squirrel monkey, in which half of the cortisol was albumin bound, and the other half remained protein free. Our most appealing finding was that in most species, as much as 68% of plasma CBG remained free of cortisol under physiologic conditions. These results are discussed with respect to the theories concerning the role of CBG in plasma transport and the local delivery of cortisol and free CBG as a proper hormone.     

Contribution of æ1-acid glycoprotein to plasma protein binding of some basic antimicrobials in pigs

Protein binding kinetics of basic antimicrobials including trimethoprim (TMP), erythromycin (EM), lincomycin (LM) and clindamycin (CM) were studied using porcine plasma, albumin and æ-acid glycoprotein (AGP). Rosenthal plots of these basic drugs in porcine plasma suggest saturable and non-saturable binding. Dissociation constants (K_d) and binding capacity (B_max) for saturable binding were as follows: TMP, K_d= 8.58 μmol/L, B_max= 5.26 μmol/L; EM, k_d= 2.72 μmol/L, B_max= 3.06 μmol/L; LM, k_d= 3.96 μmol/L, B_max= 6.58 μmol/L and CM, k_d= 4.43 μmol/L, B_max= 21.7 μmol/L. The proportionality constants (B_max2/k_d2) for non-saturable binding were 0.29 in TMP, 0.52 in EM, 0.17 in LM and 3.2 in CM. The k_ds of the drugs in porcine AGP solution were determined by a fluorescence quenching method, using 1-anilino-8-naphthalene sulphonate (ANS) as a fluorescent probe: 9.51 μmol/L in TMP, 1.89 μmol/ L in EM, 4.48 μmol/L in LM and 9.69 μmol/L, in CM. Comparable kd values between porcine plasma and AGP solution indicated that AGP is a major saturable binder in porcine plasma. Binding property to porcine albumin presented linearity, showing the following proportionality constants: 0.23 in TMP, 0.38 in EM, 0.01 in LM and 0.76 in CM. The comparable proportionality constants of TMP and EM between porcine plasma and albumin solution indicate that albumin is a major non-saturable binder, whereas proportionality constants of LM and CM in albumin solution compared to those in porcine plasma were low, implying another non-saturable binder, i.e. lipoprotein. Simulation curve of drug-binding percentage vs. AGP concentrations showed that in pigs under a pathologic state, or during early growth stage with high AGP levels, AGP could be a main contributor to drug-plasma protein binding for all drugs examined. The increase of AGP from normal to pathological concentrations induced a decrease in the unbound fraction: LM > CM > EM > TMP in order of AGP contribution to drug binding. Therefore, the disposition and efficacy of basic antimicrobials which bind to AGP with high affinity could be markedly influenced by altered AGP levels, implying AGP contribution to pharmacokinetics and pharmacodynamics.     

Physiological adaptive indicators in fasted neonate broiler chicks in response to calcium gluconate injection

Four hundred and eighty mixed‐sex broiler chicks aged 3 h after hatching were allotted according to a completely random design in a 6 × 2 × 2 factorial schedule into two groups of 12 replications of 20 chicks each. The main experimental factors were fasting for 0, 6, 12, 24, 36 and 48 h after chick placement and calcium gluconate (Ca‐glu) injection (0 and 0.6 ml). Live body weight (BW) of chicks decreased linearly (Y = 43.36–0.109BW_0 h, r² = 0.876) as neonatal fasting extended. Injection of 0.6 ml Ca‐glu at 3 h post‐hatching did not affect weight loss of chicks. Yolk residuals (YR) utilized linearly (Y = 5.75–0.062YR, r² = 0.956) by 0.062 g/h in neonate fasted chicks up to 48 h, showing no effect of Ca‐glu injection. Neonatal fasting periods longer than 12 h increased liver weight (p < 0.05). The mean absolute and proportional (% of BW_0 h) breast and leg weight were reduced linearly as neonatal fasting extended (p < 0.05). Serum glucose concentration increased up to 6 h and then reduced linearly to 150 mg/dl after 48‐h fasting. The Ca‐glu treatment influenced serum glucose level for a short period up to 6 h of fasting. Serum Ca concentration sharply increased up to threefolds in the birds received Ca‐glu injection resulting in acute hypercalcemia, then decreased to the initial level after 24‐h feed withdrawal (p < 0.05). The mean serum level for creatinine, uric acid, cholesterol, HDL, albumins and total proteins significantly increased during the fasting periods of 6 to 48 h and significantly elevated in the birds receiving 0.6‐ml Ca‐glu injection compared with the non‐treated chicks (p < 0.05). It was concluded that subcutaneous administration of 0.6 ml Ca‐glu in the chick's neck did not suitably support the increased metabolic demands for glucose and calcium in feed‐deprived neonate chicks.     

Pharmacokinetics of tulathromycin after subcutaneous injection in North American bison (Bison bison)

Tulathromycin is approved for the treatment of respiratory disease in cattle and swine. It is intended for long‐acting, single‐dose injection therapy (Draxxin), making it particularly desirable for use in bison due to the difficulty in handling and ease of creating stress in these animals. The pharmacokinetic properties of tulathromycin in bison were investigated. Ten wood bison received a single 2.5 mg/kg subcutaneous injection of Draxxin. Serum concentrations were measured by liquid chromatography–mass spectrometry (LC‐MS) detection. Tulathromycin demonstrated early maximal serum concentrations, extensive distribution, and slow elimination characteristics. The mean maximum serum concentration (C_max) was 195 ng/mL at 1.04 h (t_max) postinjection. The mean area under the serum concentration–time curve, extrapolated to infinity (AUC_0–inf), was 9341 ng·h/mL. The mean apparent volume of distribution (V_d/F) and clearance (Cls/F) was 111 L/kg and 0.4 L/h/kg, respectively, and the mean half‐life (t_1/2) was 214 h (8.9 days). Compared to values for cattle, C_max and AUC_0–inf were lower in bison, while the V_d/F was larger and the t_1/2 longer. Tissue distribution and clinical efficacy studies in bison are needed to confirm the purported extensive distribution of tulathromycin into lung tissue and to determine whether a 2.5 mg/kg subcutaneous dosage is adequate for bison.     

Regulation of growth hormone expression by thyrotropin-releasing hormone through the pituitary-specific transcription factor Pit-1 in chicken pituitary

Pit-1 is a pituitary-specific POU-domain DNA binding factor, which binds to and trans-activates promoters of growth hormone- (GH), prolactin- (PRL) and thyroid stimulating hormone beta- (TSHbeta) encoding genes. Pit-1 has been identified in several mammalian and avian species. Thyrotropin-releasing hormone (TRH) is located in the hypothalamus and it stimulates TSH, GH and PRL release from the pituitary gland. In the present study, we successfully developed a competitive RT-PCR for the detection of Pit-1 expression in the chicken pituitary, that was sensitive enough to detect picogram levels of Pit-1 mRNA. Applying this method, the effect of TRH injections on Pit-1 mRNA expression was determined in the pituitary of chick embryos and growing chicks. In both 18-day-old embryos and 10-day-old male chicks the Pit-1 mRNA expression was significantly increased following TRH injection, thereby indicating that the stimulatory effects of TRH on several pituitary hormones is mediated via its effect on Pit-1 expression. Therefore, a semi-quantitative RT-PCR method was used to detect possible changes in GH levels. TRH affected the GH mRNA levels at both developmental stages. These results, combined with the data on Pit-1 mRNA expression, indicate that Pit-1 has a role in mediating the stimulatory effects of TRH on pituitary hormones like GH.     

GABA Control of GnRH Release from the Ewe Hypothalamus In Vitro: Sensitivity to Oestradiol

The present study examines the involvement of GABA_{A or B} receptors in gonadotrophin‐releasing hormone (GnRH) release in vitro and determines whether oestradiol modulates γ‐aminobutyric acid (GABA)–GnRH interaction. Within 10 min after ewe killing, hypothalamic slices were dissected and placed in oxygenated Minimum Essential Media (MEM)‐α at 4°C; within 2 h, slices were singly perifused at 37°C with oxygenated MEM‐α (0.15 ml/min), with or without oestradiol (24 pg/ml). After 4 h equilibration, fractions were collected for 4 h interposed with a 10 min exposure to specific GABA_{A or B} receptor ligands (0.1–10 mm ). The GABA_{A or B} agonists (muscimol or baclofen) did not greatly influence GnRH release. However, GnRH increased (p < 0.05) after exposure to 10 mm GABA_{A or B} antagonists (bicuculline or CGP52432, respectively). The GABA_A antagonist stimulated greater sustained GnRH release (p < 0.05) in the absence of oestradiol than in its presence. The bioactivity of the released GnRH was studied using a hypothalamus‐pituitary sequential double‐chamber perifusion. Only after exposure of hypothalamic slices to the GABA_A antagonist, did the hypothalamic eluate stimulate luteinizing hormone release from pituitary fragments (p < 0.05) confirming that the GABA_A antagonist stimulated release of biologically active GnRH. In summary, GnRH release from the hypothalamus is predominantly under GABA_A receptor inhibitory control and this is attenuated in the presence of oestradiol.     

Diurnal changes in specific binding of cortisol to cytosolic and nuclear fractions from equine peripheral blood mononuclear cells

Recent research suggests that the subcellular location and nuclear concentration of glucocorticoid receptors may have a significant impact on cell sensitivity to stimuli acting through cyclic AMP or protein kinase C second messengers. The present study was performed in order to determine whether there may be diurnal fluctuations in the cytosolic and/or nuclear glucocorticoid receptors in equine peripheral blood mononuclear cells (PBMC). Jugular blood samples were collected by venipuncture from four resting unconditioned horses. Samples were collected at 8 AM, 2 PM, 8 PM and 2 AM. The PBMC were harvested from Ficoll gradients. Cytosolic and nuclear fractions were recovered by centrifugation from homogenized PBMC. Protein concentrations of the resulting fractions was determined and serial dilutions incubated with 3H-cortisol in the presence or absence of excess unlabeled dexametha- sone. Saturable specific binding was linear for 75 to 200 µg of protein which was recovered from 10xl0⁵ to 100xl0⁶ equine PBMCs. This binding was eliminated in heat-treated samples. Further, specific binding of cortisol to cytosolic fractions was reduced (P<.05) at 8AM as compared to binding at 2 PM and 2 AM. Nuclear receptor binding was high during daylight hours and undetectable at 2 AM. Total cellular receptor was highest at 2 PM and lowest at 2 AM. Results from this experiment suggest that diurnal fluctuations in equine cortisol concentrations are related to changes in the subcdlular distribution ofglucocorticoid receptors in PBMCs. Changes in the total amount of receptor also suggests that there are diurnal changes in glucocorticoid receptor synthesis and/ or recycling. Thus, diurnal variations in glucocorticoid interactions with second messenger pathways may be of importance in the equine species.     

Effect of dexamethasone on the expression of atrogin‐1/MAFbx in chick skeletal muscle

Expression of atrogin‐1/MAFbx, a muscle‐specific E3 ubiquitin ligase, is high under catabolic conditions, that result in muscle atrophy. Messenger RNA (mRNA) expression of atrogin‐1/MAFbx is increased by the glucocorticoid dexamethasone in mammalian skeletal muscle. This study investigated the effects of dexamethasone on expression of atrogin‐1/MAFbx in skeletal muscle of neonatal chicks and in chick myotubes. Chicks were given a single intraperitoneal injection of dexamethasone at a concentration of 10 mg/kg body weight. Twenty‐four hours after dexamethasone administration, the Pectoralis muscle weight of chicks was decreased. mRNA expression of atrogin‐1/MAFbx in skeletal muscle of chicks was significantly increased by dexamethasone administration. Expression of other proteolytic‐related genes (20S proteasome C2 subunit, m‐calpain large subunit, and cathepsin B) in skeletal muscle of chicks was not increased by dexamethasone administration. Chick myotubes were incubated with dexamethasone (1, 10 or 100 µmol/L) for 6 h. Expression of atrogin‐1/MAFbx mRNA in chick myotubes was increased in the presence of all concentrations of dexamethasone. However, expression of other proteolytic‐related genes (20S proteasome C2 subunit, m‐calpain large subunit and cathepsin B) in chick myotubes was not affected by dexamethasone treatment. These results indicate that dexamethasone enhances atrogin‐1/MAFbx expression in chick skeletal muscle, resulting in increased muscle atrophy.     

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.     

Possibility of vancomycin-resistant enterococci transmission from human to broilers, and possibility of using the vancomycin-resistant gram-positive cocci as a model in a screening study of vancomycin-resistant enterococci infection in the broiler chick

In this study, vancomycin‐resistant enterococci (VRE) from humans and vancomycin‐resistant gram‐positive cocci (VRPC) from pigs were examined for their ability to transmit in the chick intestine (Experiment 1). A model study on the spread speed of VRPC was also estimated from chick to chick under semi‐production conditions with different administration routes (not inoculated, oral administration to a chick, sprayed on the floor) (Experiment 2). Furthermore, the disappearance of VRPC from their litter with composting processes was examined (Experiment 3). Each of six chicks was inoculated with VRPC or VRE at 1 day old in Experiment 1. All the chicks had VRPC or VRE in their glandular stomach at 22 days of age. In Experiment 2, 6 floor pens covered with sawdust were prepared and 20 chicks were allotted to each pen. The chicks were inoculated with VRPC at 1 day of age. The VRPC were detected in each group in cloacal swabs at 2 days of age (detection rate; 20–80%). And they were also detected in the not‐inoculated group. The VRPC detection rate gradually decreased, and detection was rare (0–10%) in the packing chicks (50 days old). VRPC were detected in the litter of each pen in Experiment 2. A composting process was effectively used to eliminate VRPC by the 6th week (Experiment 3).     

产蛋鸡与休产鸡肝脏,输卵管雌激素受体含量的比较

本试验选用正在产蛋和休产的来航鸡,用放射配体结合分析法,分别测定两组鸡的肝脏和输卵管胞浆和胞核受体含量。胞浆液的蛋白含量用考马斯亮蓝G250法测定;胞核DNA含量用紫外吸收法测定。试验结果表明:两组鸡的肝脏和输卵管的胞浆雌激素受体含量均无明显差异;两组鸡的肝脏和输卵管的胞核雌激素受体含量,产蛋鸡肝脏的胞核雌激素受体是休产鸡的15倍,差异显著(P<0.05),而输卵管的胞核雌激素受体差异不明显。     

Development of an in vitro oviduct epithelial explants model for studying sperm–oviduct binding in the buffalo

In this study, we developed an in vitro model for studying sperm–oviduct binding in the buffalo. Oviduct explants were prepared by overnight culture of epithelial cells in TCM‐199 medium under 5% CO₂ at 38.5 °C. Cryopreserved spermatozoa from buffalo bulls (n = 4) were incubated with the oviduct explants, and the sperm–oviduct explants complex was stained with JC‐1. The effect of sperm concentration (2, 3 and 4 million), size of the oviduct explants (<0.2, 0.2–0.3, 0.3–0.4 and >0.4 mm²) and time of incubation (1 hr and 4 hr) on binding index (BI—number of sperm bound to unit area of explants) was studied. No significant difference was observed in the BI among <0.2, 0.2–0.3 and 0.3–0.4 mm² size of explants; however, the BI decreased significantly (p < .05) when the size of explants exceeded 0.4 mm². The BI decreased significantly (p < .05) when the sperm concentration was increased to 4 million, while the duration of incubation did not have any significant effect on the BI. The interaction of bulls with explants size, sperm concentration and incubation time was not significant. The developed assay has the potential to be used as an in vitro model for studying sperm–oviduct binding in the buffalo.     

In vivo measurement of central corneal thickness in normal chicks (Gallus gallus domesticus) with the optical low-coherence reflectometer

Objective To determine the practicability and accuracy of central corneal thickness (CCT) measurements in living chicks utilizing a noncontact, high‐speed optical low‐coherence reflectometer (OLCR) mounted on a slit lamp. Animals studied Twelve male chicks (Gallus gallus domesticus). Procedures Measurements of CCT were obtained in triplicate in 24 eyes of twelve 1‐day‐old anaesthetized chicks using OLCR. Every single measurement taken by OLCR consisted of the average result of 20 scans obtained within seconds. Additionally, corneal thickness was determined histologically after immersion fixation in Karnovsky’s solution alone (20 eyes) or with a previous injection of the fixative into the anterior chamber before enucleation (4 eyes). Results Central corneal thickness measurements using OLCR in 1‐day‐old living chicks provide a rapid and feasible examination technique. Mean CCT measured with OLCR (189.7 ± 3.34 μm) was significantly lower than histological measurements (242.1 ± 47.27 μm) in eyes with fixation in Karnovsky’s solution (P = 0.0005). In eyes with additional injection of Karnovsky’s fixative into the anterior chamber, mean histologically determined CCT was 195.2 ± 8.25 μm vs. 191.9 ± 8.90 μm with OLCR. A trend for a lower variance was found compared to the eyes that had only been immersion fixed. Conclusion Optical low‐coherence reflectometry is an accurate examination technique to measure in vivo CCT in the eye of newborn chicks. The knowledge of the thickness of the chick cornea and the ability to obtain noninvasive, noncontact measurements of CCT in the living animal may be of interest for research and development of eye diseases in chick models.     

Growth Hormone Receptors in the Porcine Testis During Prepuberty

Supplementation of exogenous growth hormone (GH) during prepuberty advances onset of spermatogenesis in boars, but the mechanism of action is unknown. The present study is an investigation of the presence and characteristics of testicular growth hormone receptors (GHR). A total of 36 boars were castrated, three boars every 10 days, between the ages of 10 and 120 days. Testicular membrane preparations of 10, 20, 30, 50, 70, 100 and 120‐day‐old boars were used to determine ¹²⁵I‐bGH binding and Scatchard analysis. Liver from a 60‐kg barrow was used for comparison. Specific ¹²⁵I‐bGH binding to testicular membrane preparations occurred in all age groups with the exception of 20‐day‐old boars at levels of 30–40% of liver binding. At 30 days of age the unlabelled bGH at 1.1 ng/tube achieved half maximal inhibition (ID₅₀). Results of Scatchard analysis indicated a single class of binding sites. Binding affinity was 2.89 × 10⁹m with a binding capacity of 12 fmole/mg membrane protein. The results from this study suggest that GH may act directly on the cells of the prepubertal boar testis.     

Histomorphometric and Progesterone Receptor Immunohistochemical Analysis in the Oviduct of Newly Hatched Chicks Treated with Follicle-Stimulating Hormone during Embryonic Development

In this study we evaluated the histomorphology and ultra-structure of the oviduct of newly hatched chicks, as well as the immunohistochemical expression of progesterone receptor (PR) in this tissue after follicle-stimulating hormone (FSH) treatment on days 13, 15 and 17 of embryonic development. Results indicated a marked difference in the histology of the oviduct of newly hatched chicks treated with FSH. Magnum mucosa from these animals presented a pseudostratified epithelium with evaginations from the lumen into the epithelium and from the latter into the stroma beneath where tubular glands are formed. In contrast magnum mucosa from control animals presented columnar epithelium with no evaginations. In magnum epithelium FSH also induced the formation of cilia and microvilli projections into the lumen as well as an increase in the wall and lumen areas and in the density of nuclei per unit-area. PR immunoreactivity was only observed in the oviduct of FSH treated animals. PR was located in the nucleus of epithelial luminal cells, mucosal stromal cells and smooth muscle cells. These findings suggest that FSH induces an adequate hormonal milieu for the cytodifferentiation and PR gene expression in the chick oviduct.     

Estimated plasma bupivacaine concentration after single dose and eight-hour continuous intra-articular infusion of bupivacaine in normal dogs

Objective— To estimate maximum plasma concentration (C_max) and time to maximum plasma (t_max) bupivacaine concentration after intra‐articular administration of bupivacaine for single injection (SI) and injection followed by continuous infusion (CI) in normal dogs. Study Design— Cross‐over design with a 2‐week washout period. Animals— Healthy Coon Hound dogs (n=8). Methods— Using gas chromatography/mass spectrometry, canine plasma bupivacaine concentration was measured before and after SI (1.5 mg/kg) and CI (1.5 mg/kg and 0.3 mg/kg/h). Software was used to establish plasma concentration–time curves and estimate C_max, T_max and other pharmacokinetic variables for comparison of SI and CI. Results— Bupivacaine plasma concentration after SI and CI best fit a 3 exponential model. For SI, mean maximum concentration (C_max, 1.33±0.954 μg/mL) occurred at 11.37±4.546 minutes. For CI, mean C_max (1.13±0.509 μg/mL) occurred at 10.37±4.109 minutes. The area under the concentration–time curve was smaller for SI (143.59±118.390 μg/mL × min) than for CI (626.502±423.653 μg/mL × min, P=.02) and half‐life was shorter for SI (61.33±77.706 minutes) than for CI (245.363±104.415 minutes, P=.01). The highest plasma bupivacaine concentration for any dog was 3.2 μg/mL for SI and 2.3 μg/mL for CI. Conclusion— Intra‐articular bupivacaine administration results in delayed absorption from the stifle into the systemic circulation with mean C_max below that considered toxic and no systemic drug accumulation. Clinical Relevance— Intra‐articular bupivacaine can be administered with small risk of reaching toxic plasma concentrations in dogs, though toxic concentrations may be approached. Caution should be exercised with multimodal bupivacaine administration because plasma drug concentration may rise higher than with single intra‐articular injection.     

Cellular coupling of potassium channels with β2 adrenoceptors in mediating myometrial relaxation in buffaloes (Bubalus bubalis)

Choudhury, S., Garg, S. K., Singh, T. U., Mishra, S. K. Cellular coupling of potassium channels with β₂ adrenoceptors in mediating myometrial relaxation in buffaloes (Bubalus bubalis). J. vet. Pharmacol. Therap. 33 , 22–27. Present study unravels the functional presence of potassium channels and their involvement in mediating β₂ adrenoceptors‐induced myometrial relaxation in buffalo myometrium. Isolated myometrial preparations exhibited rhythmic spontaneity with regular pattern of amplitude and frequency. Levcromakalim produced concentration‐dependent inhibitory effect on myometrial spontaneity and relaxant effect and the dose–response curve (DRC) was shifted towards right in the presence of glybenclamaide. In the tissues pretreated with glybenclamide, relaxant effect of albuterol was significantly (P < 0.05–0.001) lower (pD₂ = 6.94, R_max = 96.8 ± 3.3%; n = 5) compared with albuterol alone (pD₂ = 8.55, R_max = 101.1 ± 6.3%; n = 6) and the DRC was shifted to right. In the presence of tetraethyl ammonium (TEA) also, significant (P < 0.001) rightward shift of DRC of albuterol with decrease in maximal effect (pD₂ = 8.05, R_max = 71.2 ± 7.4%; n = 7 vs. control pD₂ = 8.55, R_max = 101.1 ± 6.3%; n = 6) was observed. On the other hand, 4‐aminopyridine (1 mm ) sensitized the myometrial strips and increased the amplitude and frequency/min of myometrial spontaneity but failed to significantly alter the DRC of albuterol. Results of present study suggest the functional presence of K_ATP, BK_Ca and K_V channels in buffalo myometrium, but β₂‐adrenoreceptor agonist‐induced myometrial relaxation seems to be K_ATP and BK_Ca channel‐dependent only. Further, our studies also suggest promising therapeutic potential of potassium channel modulators as tocolytics in buffaloes.     

Muscarinic Receptors in Equine Airways

The distribution of muscarinic receptors in equine airways was investigated using autoradiography. Frozen sections of tissue from six different levels in the bronchial tree, from the trachea to the distal bronchioles, were incubated in vitro with 1.5 nmol/L of the muscarinic receptor antagonist 1-[N-methyl-³H]scopolamine methyl chloride (³H-NMS). In addition, the subtype pattern of muscarinic receptors was investigated in equine tracheal smooth muscle using radioligand binding with methoctramine, tripinamide, 4-DAMP-methiodide and pirenzipine as competitors against the binding of 1.3 nmol/L ³H-NMS. The autoradiograms showed specific labelling indicating a high density of muscarinic receptors in smooth-muscle tissue in all levels of the airway tree investigated. Besides muscle tissue, subepithelial glands were the only structures specifically labelled. The dominating subtypes in tracheal smooth muscle investigated with radioligand binding studies were found to be M₂ and M₄, as both methoctramine (pK _d = 8.5) and tripinamide (pK _d = 8.6 and 6.7 for two different sites) showed high affinity. The density of the M₃-muscarinic receptor subtype was low, but this subtype could be detected with statistical significance when methoctramine was used as the competitor against ³H-NMS binding.     

The effects of l‐DOPA and sulpiride on growth hormone secretion at different injection times in Holstein steers

The effects of l ‐DOPA, a precursor of dopamine (DA), and sulpiride, a D₂‐type DA receptor blocker, on growth hormone (GH) and prolactin (PRL) secretion were investigated in steers. Eight Holstein steers (212.8 ± 7.8 kg body weight) were used. Lighting conditions were 12:12 L:D (lights on: 06.00–18.00 hours). Blood samplings were performed during the daytime (11.00–15.00 hours) and nighttime (23.00–03.00 hours). Intravenous injections of drugs or saline were performed at 12.00 hour for the daytime and 00.00 hour for the nighttime, respectively. Plasma GH and PRL concentrations were determined by radioimmunoassay. l ‐DOPA did not alter the GH secretion when it was injected at 12.00 hour (spontaneous GH level at its peak). On the other hand, l ‐DOPA increased GH secretion at 00.00 hour (GH level at its trough). Injection of sulpiride suppressed GH secretion at 12.00 hour but did not affect GH levels at 00.00 hour. l ‐DOPA inhibited and sulpiride stimulated PRL release during both periods. These results suggest that dopaminergic neurons have stimulatory action on GH secretion and inhibitory action on PRL secretion in cattle. In addition, injection time should be considered to evaluate the exact effects on GH secretion due to its ultradian rhythm of GH secretion in cattle.