EP2A promotes human CD34+ cell homing in vitro but not proliferation

AIM: To investigate the role of prostaglandin E₂ receptor 2 agonist (EP₂A) in proliferation and homing of human CD34⁺ cells. METHODS: Bone marrow fluid and peripheral blood containing stem cells were collected from healthy donors mobilized by granulocyte colony-stimulating factor in our department. Human CD34⁺ cells were isolated by the method of magnetic-activated cell sorting microbeads. Bone marrow mononuclear cells were isolated by Ficoll-Paque centrifugation, and the bone marrow mesenchymal stem cells (BMMSC) were cultured with L-DMEM. Human CD34⁺ cells and BMMSC were divided into 4 groups, and treated with PGE₂ (as positive control), DMSO (as negative control), EP₂A and EP₂A+prostaglandin E₂ receptor 2 antagonist (EP₂AA), respectively. After exposed to the reagents, human CD34⁺ cell viability was measured by CCK-8 assay, the number of colonies was evaluated by colony-formation assay, the cell cycle distribution was analyzed by flow cytometry, and the protein expression of survivin, β-catenin and CXC chemokine receptor 4 (CXCR4) was detrmined by Western blot. Moreover, the concentration of stromal cell-derived factor-1α (SDF-1α) in the BMMSC was detected by ELISA. RESULTS: The cell viability and the colony number of human CD34⁺ cells in EP₂A group were not higher than those in negative control group. Furthermore, the proportion of human CD34⁺ cells treated with EP₂A in G₂/M phase was not elevated compared with negative control group. The protein expression of survivin and β-catenin did not up-regulated in human CD34⁺ cells exposed to EP₂A, but the protein expression of CXCR4 in human CD34⁺ cells and the concentration of SDF-1α in BMMSC were elevated. CONCLUSION: EP₂A promotes human CD34⁺ cell homing in vitro but not proliferation.     

苯乙醇胺A及其残留检测方法研究进展

苯乙醇胺A作为一种新型的β-兴奋剂,具有促进动物蛋白质合成、提高饲料转化率的作用,但其在动物性食品中的残留物对人体健康构成威胁,故近年来引起各界学者的广泛关注。因此建立便捷高效的检测方法至关重要。主要介绍了苯乙醇胺A的理化性质、药理作用、代谢规律、毒理研究以及其在不同样品基质中残留检测的方法,对研究方法的开发与研究提供有益帮助。     

Pharmacokinetics of morphine in combination with dexmedetomidine and maropitant following intramuscular injection in dogs anaesthetized with halothane

The purpose of this study was to evaluate the pharmacokinetics of morphine in combination with dexmedetomidine and maropitant injected intramuscularly in dogs under general anaesthesia. Eight healthy dogs weighing 25.76 ± 3.16 kg and 3.87 ± 1.64 years of age were used in a crossover study. Dogs were randomly allocated to four groups: (1) morphine 0.6 mg/kg; (2) morphine 0.3 mg/kg + dexmedetomidine 5 μg/kg; (3) morphine 0.3 mg/kg + maropitant 1 mg/kg; (4) morphine 0.2 mg/kg + dexmedetomidine 3 μg/kg + maropitant 0.7 mg/kg. Blood samples were collected before, 15 and 30 min, and 1, 2, 3 4, 6 and 8 hr after injection of the test drugs. Plasma concentration of the drugs was determined by liquid chromatography-mass spectrometry. The elimination half-life (T_1/2) of morphine was higher and the clearance rate (CL) was lower when combined with dexmedetomidine (T_1/2 = 77.72 ± 20.27 min, CL = 119.41 ± 23.34 ml kg⁻¹ min⁻¹) compared to maropitant (T_1/2 = 52.73 min ± 13.823 ml kg⁻¹ min⁻¹, CL = 178.57 ± 70.55) or morphine alone at higher doses (T_1/2 = 50.53 ± 12.55 min, CL = 187.24 ± 34.45 ml kg⁻¹ min⁻¹). Combining morphine with dexmedetomidine may increase the dosing interval of morphine and may have a clinical advantage.     

An update on cardiovascular adrenergic receptor physiology and potential pharmacological applications in veterinary critical care

Objective: To review the recent human and veterinary literature on current adrenergic receptor physiology/pathophysiology and potential applications in veterinary critical care. Data sources: Human and veterinary clinical studies, reviews, texts, and recent research in receptor molecular biology. Human data synthesis: Recent development of molecular cloning and other biological research techniques has advanced the field of adrenergic physiology. The past decade of research has made available new knowledge of adrenergic receptor subtypes as well as their locations and functions. Many of the diagnostic compounds used in biochemical research to distinguish between α‐ and β‐receptor subtypes may emerge as important additions to the arsenal of cardiovascular pharmaceuticals. Veterinary data synthesis: Veterinary adrenoceptor research is typically directed at investigating the effects of commercially available medications. Such studies demonstrate important species differences in addition to potential side effects and new indications for therapy. Many of the human molecular biology studies are performed on animal species, which can have direct application to veterinary medicine. Conclusions: Proper cardiovascular responses are essential to maintaining tissue perfusion and cellular homeostasis. α‐ and β‐adrenergic receptors play a vital role not only in the pathophysiology but also in the therapy of diseases involving the cardiovascular system. For adrenergic pharmacotherapy to be successfully used, a thorough understanding of the mechanisms underlying adrenoceptor physiology is necessary. Recent research has illuminated various subsets within the α‐ and β‐receptor classifications. Awareness of currently available and emerging adrenoceptor subtype‐specific treatment options allows precise pharmacologic targeting of disease processes in critical illness.     

Pharmacokinetics and bioavailability after intramuscular injection of the 5‐HT1A serotonin agonist R‐8‐hydroxy‐2‐(di‐n‐propylamino) tetralin (8‐OH‐DPAT) in domestic goats (Capra aegagrus hircus)

To determine the bioavailability and pharmacokinetic properties of the serotonin 5‐HT_1A receptor agonist R‐8‐OH‐DPAT in goats, and 0.1 mg kg^?1 R‐8‐OH‐DPAT hydrobromide was administered intramuscularly (i.m.) and intravenously (i.v.) to six goats in a two‐phase cross‐over design experiment. Venous blood samples were collected from the jugular vein 2, 5, 10, 15, 20, 30, 40 and 60 min following treatment and analysed by liquid chromatography tandem mass spectrometry. Bioavailability and pharmacokinetic parameters were determined by a one‐compartment analysis. Mean bioavailability of R‐8‐OH‐DPAT when injected i.m. was 66%. The mean volume of distribution in the central compartment was 1.47 L kg^?1. The mean plasma body clearance was 0.056 L kg^?1 min^?1. All goats injected i.v. and two of six goats injected i.m. showed signs of serotonin toxicity. In conclusion, R‐8‐OH‐DPAT is well absorbed following i.m. injection and the observed pharmacokinetics suggest that administration via dart is feasible. Administration of R‐8‐OH‐DPAT hydrobromide, at a dosage of 0.1 mg kg^?1, resulted in the observation of clinical signs of serotonin toxicity in the goats. It is suggested that dosages for the clinical use of the compound should be lower in order to achieve the desired clinical effect without causing serotonin toxicity.     

A possible solution to model nonlinearity in elimination and distributional clearances with α2‐adrenergic receptor agonists: Example of the intravenous detomidine and methadone combination in sedated horses

The alpha(α)₂‐agonist detomidine is used for equine sedation with opioids such as methadone. We retrieved the data from two randomized, crossover studies where detomidine and methadone were given intravenously alone or combined as boli (STUDY 1) (Gozalo‐Marcilla et al., 2017, Veterinary Anaesthesia and Analgesia, 2017, 44 , 1116) or as 2‐hr constant rate infusions (STUDY 2) (Gozalo‐Marcilla et al., 2019, Equine Veterinary Journal, 51 , 530). Plasma drug concentrations were measured with a validated tandem Mass Spectrometry assay. We used nonlinear mixed effect modelling and took pharmacokinetic (PK) data from both studies to fit simultaneously both drugs and explore their nonlinear kinetics. Two significant improvements over the classical mammillary two‐compartment model were identified. First, the inclusion of an effect of detomidine plasma concentration on the elimination clearances (Cls) of both drugs improved the fit of detomidine (Objective Function Value [OFV]: ?160) and methadone (OFV: ?132) submodels. Second, a detomidine concentration‐dependent reduction of distributional Cls of each drug further improved detomidine (OFV: ?60) and methadone (OFV: ?52) submodel fits. Using the PK data from both studies (a) helped exploring hypotheses on the nonlinearity of the elimination and distributional Cls and (b) allowed inclusion of dynamic effects of detomidine plasma concentration in the model which are compatible with the pharmacology of detomidine (vasoconstriction and reduction in cardiac output).     

Dexmedetomidine-induced pulmonary alterations in sheep

Alpha(2) agonist-induced pulmonary oedema in sheep might be related to alterations in pulmonary haemodynamics and/or activation of inflammatory processes. In seven sevoflurane-anaesthetized sheep pulmonary haemodynamics, arterial oxygen tensions, nitric oxide and prostaglandin E(2) concentrations were determined before and after intravenous dexmedetomidine (2microg kg(-1)). In a second trial, lung tissue was sampled for histopathology and quantitative real-time PCR for IL-1beta and iNOS mRNA in a control sheep and 2, 10 and 30min after dexmedetomidine. Computer tomography of the lung under sevoflurane anaesthesia before and after dexmedetomidine was performed. Two minutes after dexmedetomidine mean pulmonary artery pressure, pulmonary arterial occlusion pressure and estimated capillary pressurewere significantly increased to 34.5mmHg, 22.2mmHg and 27.1mmHg, respectively. On computer tomography, lung density increased immediately after dexmedetomidine, with maximal density occurring between 9 and 12min. Histopathology was consistent with vascular congestion followed by protein and erythrocyte extravasation into alveoli. Increased iNOS mRNA levels were detected in sevoflurane anaesthetized animals only. An IL-1beta signal occurred after morphological changes had occurred in lung tissue. These findings support hydrostatic stress as the underlying cause of alpha(2) agonist-induced pulmonary oedema in sheep.     

Comparison of the effects of the alpha-2 agonists detomidine, romifidine and xylazine on nociceptive withdrawal reflex and temporal summation in horses

ObjectiveTo evaluate and compare the antinociceptive effects of the three alpha-2 agonists, detomidine, romifidine and xylazine at doses considered equipotent for sedation, using the nociceptive withdrawal reflex (NWR) and temporal summation model in standing horses.Study designProspective, blinded, randomized cross-over study.AnimalsTen healthy adult horses weighing 527–645 kg and aged 11–21 years old.MethodsElectrical stimulation was applied to the digital nerves to evoke NWR and temporal summation in the left thoracic limb and pelvic limb of each horse. Electromyographic reflex activity was recorded from the common digital extensor and the cranial tibial muscles. After baseline measurements a single bolus dose of detomidine, 0.02 mg kg^?1, romifidine 0.08 mg kg^?1, or xylazine, 1 mg kg^?1, was administered intravenously (IV). Determinations of NWR and temporal summation thresholds were repeated at 10, 20, 30, 40, 60, 70, 90, 100, 120 and 130 minutes after test-drug administration alternating the thoracic limb and the pelvic limb. Depth of sedation was assessed before measurements at each time point. Behavioural reaction was observed and recorded following each stimulation.ResultsThe administration of detomidine, romifidine and xylazine significantly increased the current intensities necessary to evoke NWR and temporal summation in thoracic limbs and pelvic limbs of all horses compared with baseline. Xylazine increased NWR thresholds over baseline values for 60 minutes, while detomidine and romifidine increased NWR thresholds over baseline for 100 and 120 minutes, respectively. Temporal summation thresholds were significantly increased for 40, 70 and 130 minutes after xylazine, detomidine and romifidine, respectively.Conclusions and clinical relevanceDetomidine, romifidine and xylazine, administered IV at doses considered equipotent for sedation, significantly increased NWR and temporal summation thresholds, used as a measure of antinociceptive activity. The extent of maximal increase of NWR and temporal summation thresholds was comparable, while the duration of action was drug-specific.     

Effects of fenoldopam mesylate on systemic hemodynamics and indices of renal function in normotensive neonatal foals

BACKGROUND: Fenoldopam mesylate, a dopamine-1 receptor agonist, has dose- and species-dependent effects on hemodynamics and renal function. The effects of this drug in normotensive neonatal foals have not been reported. HYPOTHESIS: Two doses of fenoldopam would result in distinct changes in the systemic circulation, urine output, and creatinine clearance of neonatal foals. ANIMALS: Six Thoroughbred foals. METHODS: Each foal received 2 dosages of fenoldopam (low dose, 0.04 microg/kg/min; high dose, 0.4 microg/kg/min) and a control administration of saline, in a masked, placebo-controlled study. RESULTS: High-dosage fenoldopam had no effect on renal function but caused a significant increase in heart rate and decrease in mean, systolic and diastolic arterial blood pressure compared with saline. Low-dosage fenoldopam had no effects on systemic hemodynamics, significantly increased urine output, and had no significant effect on creatinine clearance or the fractional excretions of sodium, potassium, or chloride compared with saline. CONCLUSIONS AND CLINICAL IMPORTANCE: These data suggest that high-dosage fenoldopam increases heart rate, decreases arterial blood pressure, and has no significant effects on renal function, whereas low-dosage fenoldopam has no significant effects on systemic hemodynamics while increasing urine output. This contrast is unique to this species and warrants further investigation.     

Comparison of lidocaine, xylazine, and lidocaine−xylazine for caudal epidural analgesia in cattle

Objective To directly compare the time to onset and duration of analgesia produced by a lidocaine/xylazine combination with that produced by lidocaine and xylazine administered alone in the caudal epidural space of dairy cattle. Design Prospective randomized experimental study. Animals Nine adult (> 4 years of age) dairy cows (520–613 kg). Methods Caudal epidural analgesia was produced in all cows with 2% lidocaine (0.22 mg kg^?1; 5.5 mL 500 kg^?1), 10% xylazine (0.05 mg kg^?1 diluted to 5.5 mL 500 kg^?1 with sterile water), and 2% lidocaine/10% xylazine (0.22 mg kg^?1/0.05 mg kg^?1; total volume of 5.7 mL 500 kg^?1), at no earlier than weekly intervals in a Latin square design. Time to onset, duration and cranial spread of analgesia were recorded, as were degree of sedation, ataxia and ptyalism. Results No significant difference (p > 0.05) was noted for time (mean ± SEM) of onset of analgesia between lidocaine (4.8 ± 1.0 minutes) and the lidocaine/xylazine combination (5.1 ± 0.9 minutes) but onset of analgesia following xylazine was significantly longer (11.7 ± 1.0 minutes) than either of the other two treatments. Lidocaine/xylazine (302.8 ± 11.0 minutes) produced analgesia of significantly longer duration than that of xylazine (252.9 ± 18.9 minutes) and both the lidocaine/xylazine combination and xylazine alone produced analgesia of significantly longer duration than that produced by lidocaine (81.8 ± 11.8 minutes). In all cattle, xylazine, administered either alone or with lidocaine, induced mild to moderate sedation and ataxia and cutaneous analgesia from the coccyx to T13. Mild ataxia was also present in those cattle receiving lidocaine alone. Conclusion The combination of xylazine and lidocaine produces analgesia of quicker onset and longer duration than xylazine administered alone and of longer duration than lidocaine administered alone. Clinical relevance Utilizing this combination, long‐duration obstetrical and surgical procedures could commence relatively soon after epidural injection and could be completed without re‐administration of anesthetic agents.