Digoxin- and monensin-induced changes of intracellular Ca2+ concentration in isolated guinea-pig ventricular myocyte.

This study was undertaken to determine the possible mechanisms of actions of monensin and digoxin by using isolated guinea-pig ventricular myocytes. Since Ca2+ is the major signal for triggering contraction of cardiac muscle, the objective of this study was to determine whether monensin and digoxin affect the [Ca2+]i of cardiac myocytes and if so is this effect due to an increase in [Na+]i. Three different concentrations of digoxin (0.3, 1 and 3 micromol/l) and three different concentrations of monensin (0.3, 1 and 3 micromol/l) were used. Each treatment was monitored for two hours by using computerized fluoroscopy. Both digoxin and monensin increased the [Ca2+]i and accelerated the onset time of [Ca2+]i increase in a dose-dependent manner. Normal myocytes (loaded with fura-2 for 30 min before the treatment) were also compared with 'weakened' myocytes (loaded with fura-2 for 3 h before the treatment to create a 'weakened' condition). It was found that although 0.3 micromol/l monensin and digoxin did not change the [Ca2+]i in normal myocytes, they increased the [Ca2 +]i in 'weakened' myocytes. Finally, a Na+-free medium was used to demonstrate the effect of [Na+]o on both monensin- and digoxin-induced increases in [Ca2+]i. It was found that digoxin did not increase the [Ca2+]i in the Na+-free medium. Although monensin increased the [Ca2+]i in the Na+-free solution, this increase was not as large as in the Na+-containing medium. The results of the study led to the conclusion that the positive inotropic effect of digoxin depends on [Na+]o. However, monensin increases [Ca2+]i in Na+-dependent and -independent ways. An addition conclusion was that 'weakened' myocytes are more sensitive to the monensin and digoxin treatment than normal myocytes.

Functional changes in isolated guinea-pig papillary muscle induced by monensin and digoxin.

The effects of digoxin and monensin on contraction force (CF), initial contraction velocity (ICV), average contraction velocity (ACV), initial relaxation velocity (IRV) and stimulus to response time (ST) in 'fatigued' (tired) and 'non-fatigued' (fresh) guinea-pig papillary muscles were investigated. 'Fatigued' muscles had lost 30% of their original CF with the elapse of time before they were treated. The 5 h of measurement were divided into five periods (T0 was equilibration, T1, T2, T3 and T4 were, respectively, 1, 2, 3 and 4 h after drug administration). It was found that both monensin and digoxin increased the CF, ICV and ACV at T1 and increased the IRV at T2. Digoxin lost its effect with the elapse of time while monensin did not. Digoxin also decreased the ST at T2, T3 and T4. However, monensin did not change the ST. It was also found that 'fatigued' and 'non-fatigued' guinea-pig papillary muscles did not respond to the drug treatment differently. It was concluded that the initial effects of these two drugs on guinea-pig papillary muscles are similar regarding contractility but in time digoxin loses its effect while monensin does not.

Retrograde flow of spermatozoa into the urinary bladder of boars during collection of semen by electroejaculation.

Boars that had a catheter implanted in the urinary bladder (n = 11) were used to determine the magnitude of retrograde flow of spermatozoa into the urinary bladder during electroejaculation. The overall mean (+/- SD) number of spermatozoa in the electroejaculate of boars was 22 +/- 20 x 10(9), with a mean range for individual boars of 3 +/- 3 to 48 +/- 13 x 10(9). The overall mean adjusted total number of spermatozoa in the post-electroejaculation urine was 1.038 +/- 2.656 x 10(9), and the mean percentage of retrograde flow of spermatozoa into the urinary bladder among boars ranged from 0% to 32.69%, with an overall mean percentage of retrograde flow of 7.51 +/- 17.82%. These findings indicate that in boars electroejaculation is associated with retrograde flow of spermatozoa into the bladder.

Urinary losses of spermatozoa during ejaculation are negligible in boars.

Boars that had a catheter implanted surgically in the urinary bladder (n = 10) were used to determine the magnitude of retrograde flow of spermatozoa into the urinary bladder during ejaculation (Experiments 1 and 2) and the post-ejaculatory retention of spermatozoa in the urethra (Experiment 2). The overall mean (+/- SD) total number of spermatozoa in the ejaculates of boars used in Experiments 1 and 2 was 62 +/- 25 x 10(9) and 65 +/- 33 x 10(9), respectively. The overall mean adjusted total number of spermatozoa in the post-ejaculation urine of boars was 106 +/- 537 x 10(6) in Experiment 1, and 41 +/- 242 x 10(6) in Experiment 2. The overall mean percentage of retrograde flow of spermatozoa into the urinary bladder was 0.15 +/- 0.78% for the boars used in Experiment 1, and 0.03 +/- 0.16% for boars used in Experiment 2. In Experiment 2, the overall mean percentage of urethral loss of spermatozoa was 0.45 +/- 1.02%, and the overall mean percentage of total urinary losses was 0.48 +/- 1.03%. These findings demonstrate that in boars, in contrast to bulls, rams, dogs, and cats, urinary losses of spermatozoa during ejaculation are negligible.

Effect of R(-)2,5-dimethoxy-4-methylamphetamine on uterine and umbilical blood flow in conscious pregnant sheep.

Pregnant sheep and their fetuses were instrumented between 110 to 120 days of gestation (term, 145 days) for monitoring maternal and fetal arterial blood pressure, heart rate and blood flow in the maternal uterine and fetal intra-abdominal umbilical arteries. The administration of 2,5-dimethoxy-4-methylamphetamine (DOM, a 5-HT2 agonist) i.v. to the ewe in doses ranging from 1 to 20 micrograms/kg of ewe body weight produced dose-dependent decreases in the blood flow of the uterine and umbilical arteries. This was accompanied by an increase in the arterial blood pressure of the mother and fetus and a decrease in the fetal heart rate. DOM significantly increased the vascular resistance to blood flow in the uterine and umbilical arteries. The maximal increase in the vascular resistance of the uterine and umbilical arteries was 19.6- and 2.6-fold, respectively. Ketanserin, a 5-HT2 antagonist (1 mg/kg), administrated 30 min prior to DOM significantly inhibited the reduction in blood flow in the uterine and umbilical arteries to DOM and blocked the increased vascular resistance in these vessels. The inhibitory effects of ketanserin on the responses to DOM in the uterine and umbilical arteries were surmountable. Our results indicate that DOM is a potent constrictor of the uterine and umbilical vasculature which may lead to fetal distress as evidenced by a decrease in fetal heart rate and arterial blood PO2. 5-HT2 receptor stimulation by DOM may be involved in these effects since they were blocked by ketanserin.

In vitro effects of cyclopiazonic acid mycotoxin on turkey papillary muscles.

An in vitro bioassay system was used to study the effects of cyclopiazonic acid (CPA) mycotoxin on cardiac muscle. Acute exposure to 6 micrograms of CPA/ml of modified Krebs-Henseleit solution significantly (P less than 0.05) decreased 5 in vitro turkey cardiac muscle performance criteria: maximal weight a muscle could lift; maximal contraction velocity; relaxation velocity; time to peak contraction; and total time for muscle contraction and relaxation. The effect on these 5 criteria appeared to result from intracellular changes partially associated with calcium availability and were irreversible, suggesting that physiologic changes had developed after acute exposure to CPA.

[Effects of xylazole on heart rate and blood pressure in conscious dogs].

Xylazole (Xyl) is an analogue of xylazine (Xyn) synthesized by Lanzhou Institute of Chinese Traditional Veterinary Medicine. The effects of Xyl on heart rate and blood pressure were studied in 5 conscious dogs. Xyl 1 mg/kg iv was similar to Xyn in producing bradycardia and an initial transient hypertension followed by a lasting hypotension which was less significant than Xyn. Yohimbine (0.1 and 0.3 mg/kg), an alpha 2-adrenoreceptor blocking agent, antagonized bradycardia and hypotension induced by Xyl. Tolazoline (3.3 mg/kg), a nonselective alpha-adrenoreceptor blocking agent, reversed the bradycardia and hypotensive effect. Prazosin (1 mg/kg), an alpha 1-adrenoreceptor blocking agent, did not change Xyl-induced bradycardia and hypotension. Atropine (20 micrograms/kg) not only antagonized Xyl-induced bradycardia but also changed from bradycardia to tachycardia, and greatly potentiated Xyl-induced hypertension for more than 30 min. The results suggested that Xyl-induced cardiovascular effects are similar to Xyn that mediated by alpha 2-adrenoreceptor.

Effects of idazoxan, tolazoline, and yohimbine on xylazine-induced respiratory changes and central nervous system depression in ewes.

We compared the ability of 3 alpha 2-adrenoreceptor antagonists, idazoxan (0.05 mg/kg), tolazoline (2 mg/kg), and yohimbine (0.2 mg/kg) to reverse xylazine (0.3 mg/kg)-induced respiratory changes and CNS depression in 6 ewes. Once weekly, each ewe was given a random IV treatment of xylazine, followed in 5 minutes by either an antagonist or 0.9% NaCl solution. Xylazine alone caused recumbency for 54.2 +/- 5.3 minutes (mean +/- SEM). Xylazine also increased respiratory rate and decreased PaCO2 for at least 45 minutes, but did not significantly change arterial pH or PaCO2. Idazoxan and tolazoline were equally effective in reversing the respiratory actions of xylazine; however, yohimbine was less effective in reducing the respiratory rate and was ineffective in antagonizing the decreased PaO2. Idazoxan and tolazoline decreased the duration of xylazine-induced recumbency to 6.3 +/- 0.6 and 9.5 +/- 2.3 minutes, respectively, whereas yohimbine did not significantly change this effect of xylazine. Thus, at the dosages studied, idazoxan and tolazoline appeared to be more effective than yohimbine in reversing the respiratory and CNS depressant actions of xylazine in sheep.

The effects of jingsongling, a xylazine analog, on mean arterial blood pressure and heart rate in dogs--influences of yohimbine, tolazoline, prazosin, and atropine.

The effects of jingsongling (JSL) and xylazine on heart rate (HR) and mean arterial pressure (MAP) were studied in five conscious male dogs. An i.v. injection of xylazine (1 mg/kg) caused a bradycardia, an initial hypertension, and a subsequent hypotension. An i.v. injection of JSL (1 mg/kg) caused a bradycardia and a 20-min hypertension without a subsequent hypotension. Atropine sulfate (45 micrograms/kg, i.v.) increased HR for 30 min without changing MAP, and antagonized JSL-induced bradycardia for at least 60 min. There was a subsequent rebound bradycardia. Atropine sulfate potentiated JSL-induced hypertension in both magnitude and duration. Yohimbine (0.1 mg/kg, i.v.), an alpha 2-adrenoceptor antagonist, increased HR and MAP for 110 and 70 min, respectively. Yohimbine not only failed to potentiate but even reversed the pressor effect of JSL in a dose-dependent manner. Yohimbine also caused a dose-dependent reversal of JSL-induced bradycardia. Tolazoline (5 mg/kg, i.v.), a nonselective alpha-adrenoceptor antagonist, increased MAP for 20 min without changing HR. Tolazoline also reversed JSL-induced hypertension and bradycardia. Prazosin (1 mg/kg), an alpha 1-adrenoceptor antagonist, decreased MAP and increased HR for at least 110 min. Prazosin reversed JSL-induced hypertension but failed to affect JSL-induced bradycardia. These results indicated that: (1) JSL-induced bradycardia and hypertension are mediated by alpha 2-adrenoceptors; (2) yohimbine and tolazoline may be useful in antagonizing these untoward reactions associated with JSL administration, whereas prazosin and atropine were not found to be beneficial in this regard.

In vitro evaluation of isolated turkey cardiac muscle function.

Turkey papillary muscles can function in an isolated chamber containing oxygenated, pH-regulated, modified Krebs-Henseleit solutions. Experiments were conducted for up to 6 hr without dimunition of muscle function. Our data indicate that different calcium concentrations affect muscle contraction and relaxation velocities, load values, and latencies. This controlled in vitro biological assay system can be used in further studies to evaluate species specific cardiac toxins or drugs.

The influence of atropine on the cardiopulmonary effects of a xylazine-ketamine combination in dogs.

The effect of atropine on cardiopulmonary changes induced by xylazine-ketamine anesthesia was studied in 7 male dogs. A consecutive i.v. injection of xylazine (1.1 mg/kg) and ketamine (11 mg/kg) induced an anesthesia of approximately 20 min, and increased PaCO2 but decreased PaO2 and pHa for 20 min. The xylazine-ketamine administration increased heart rate for 5 min, which was followed by a small but significant decrease at 30 min posttreatment. The xylazine-ketamine injection decreased left ventricular and mean aortic pressure for 1 to 2 min. The low pressures returned to normal and became higher than base line levels at 5 min posttreatment. The xylazine-ketamine injection increased peripheral resistance and decreased cardiac output and cardiac index for at least 40 min. An i.v. injection of atropine sulfate (0.04 mg/kg) potentiated the effect of xylazine-ketamine on PaCO2 and PaO2, but did not change the effect of xylazine-ketamine on pHa. The atropine injection did not change the transient low left ventricular and mean aortic pressures immediate after the xylazine-ketamine administration, but did potentiate the duration of the increased heart rate, left ventricular and mean aortic pressures induced by xylazine-ketamine between 5 to 40 min. Atropine did not change the effect of xylazine-ketamine on peripheral resistance. Atropine partially antagonized the effect of xylazine-ketamine on cardiac output and cardiac index. Interpretation of the results indicates that atropine may increase cardiac work and may potentiate the pulmonary effect of i.v. administration of xylazine-ketamine.

Retrograde flow of spermatozoa into the urinary bladder of rams.

Retrograde flow of spermatozoa into the urinary bladder of rams during electroejaculation (EE) was examined. In experiment 1, semen and 4 consecutive samples of the urine released during the first post-EE micturition were collected once a week from 6 rams for 5 weeks during the nonbreeding season. The overall mean concentration per milliliter and the mean total number of spermatozoa in the urine ranged from 3.06 to 4.32 X 10(6) and from 80 to 2,865 X 10(6), respectively. The spermatozoal concentration in sequential urine samples was not different between samples, indicating that these spermatozoa had mixed with the urine before micturition. The percentage of the total number of spermatozoa displaced during EE, which flowed into the urinary bladder (retrograde flow), varied among rams (range 3.9% to 80%). The overall mean percentage of retrograde flow during the nonbreeding season was 28.3%. In experiment 2, a catheter was implanted into the urinary bladder of 6 rams (4 rams were from experiment 1), and semen was collected over 4 weeks during the subsequent breeding season. A urine sample was collected from the implanted catheter before EE. Immediately after semen collection, urine was collected by evacuating the bladder. The spermatozoal concentration in the pre-EE urine ranged from 0 to 1.3 X 10(6) (mean +/- SD, 0.17 +/- 0.38 X 10(6)) and was significantly (P less than 0.0001) lower than the spermatozoal concentration in the post-EE urine, which ranged from 1.10 to 22.55 X 10(6) (mean +/- SD, 9.46 +/- 11.30 X 10(6)).(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of amitraz on heart rate and aortic blood pressure in conscious dogs: influence of atropine, prazosin, tolazoline, and yohimbine.

The effect of amitraz on heart rate (HR) and mean aortic blood pressure (MAP) were studied in five conscious male dogs. An iv injection of amitraz (1 mg/kg) caused a decrease in HR, which was accompanied by sinus arrhythmia for at least 60 min. Administration of amitraz also caused an increase in MAP for 20 min. Atropine sulfate (0.045 mg/kg, iv) increased HR and prevented amitraz-induced bradycardia. In addition, atropine potentiated amitraz-induced hypertension for 45 min. Yohimbine, an alpha 2-adrenoreceptor antagonist, given iv at 0.1 mg/kg, prevented hypertension, bradycardia, and sinus arrhythmia induced by amitraz. Tolazoline, a nonselective alpha-adrenoreceptor antagonist, given iv at 5 mg/kg, reduced the bradycardia and sinus arrhythmia caused by amitraz administration but did not change amitraz-induced hypertension. Tolazoline alone also increased both HR and MAP. Prazosin, an alpha 1-adrenoreceptor antagonist, given iv at 1 mg/kg, did not affect the cardiovascular actions of amitraz. The results suggest that (1) alpha 2-adrenoreceptors mediate amitraz-induced bradycardia and hypertension, and (2) yohimbine, but not atropine, can be used to control the untoward reactions of amitraz.

Effect of xylazine on heart rate and arterial blood pressure in conscious dogs, as influenced by atropine, 4-aminopyridine, doxapram, and yohimbine.

The effects of xylazine on heart rate (HR) and mean arterial blood pressure (ABP) were studied in 5 conscious male dogs. An IV injection of xylazine (1 mg/kg) caused a decrease in HR, which was accompanied by sinus arrhythmia. Xylazine administration also caused an initial increase in ABP, which was followed by a decrease. Atropine sulfate (0.045 mg/kg, IM) increased both the ABP and HR, but prevented xylazine-induced bradycardia only in 3 of 5 dogs. The other 2 dogs had to be given a supplemental dose of atropine sulfate (0.01 mg/kg, IV) before xylazine-induced bradycardia was antagonized. In addition, atropine sulfate potentiated xylazine-induced hypertension for 60 minutes. Yohimbine, an alpha 2-adrenoreceptor blocking agent, given IV at a dosage of 0.1 mg/kg, antagonized hypertension, hypotension, and bradycardia induced by xylazine. In addition, doxapram HCl, given IV at a dosage of 5.5 mg/kg, antagonized bradycardia but potentiated xylazine-induced hypertension, and an IV injection of 4-aminopyridine at a dosage of 0.5 mg/kg did not affect the cardiovascular actions of xylazine. It was concluded that atropine sulfate at the IM dosage of 0.045 mg/kg may be insufficient to antagonize xylazine-induced bradycardia but may potentiate xylazine-induced hypertension, and yohimbine may be useful in antagonizing these untoward reactions associated with xylazine administration. Doxapram and 4-aminopyridine were not found to be beneficial.

Intravenous glucose tolerance test in cats: influenced by acetylpromazine, ketamine, morphine, thiopental, and xylazine.

Influences of chemical restraining agents of IV glucose tolerance test were studied in 3 cats. An IV injection of 0.5 g of glucose/kg produced hyperglycemia which was apparent at 15 minutes after the injection was done and disappeared at 45 minutes. All of the following drugs studied influenced the IV glucose tolerance test (0.5 g of glucose/kg): thiopental sodium (16 mg/kg, IV), ketamine (11 and 33 mg/kg, IM), acetylpromazine (2.2 mg/kg, IM), xylazine (2.2 mg/kg, IM), and morphine (0.1 mg/kg, IM). The results indicate that in cats glucose tolerance tests probably should be performed without chemical restraint. Data derived from tests where chemical restraint was used should be interpreted carefully, since the restraint drug, per se, may interfere with the test.

Fetal rat development as influenced by maternal lead exposure.

The teratogenic and neurologic effects of lead acetate on fetal rat development were investigated. Thirty-six female hooded rats were assigned to 4 treatment groups (0, 50, 75, and 100 mg/kg) and given daily oral doses of lead acetate. Animals were treated for 3 weeks prior to breeding and continuing throughout gestation. Rats were euthanized at day 20 of gestation. Blood sampling indicated that maternal blood lead concentrations in treated dams were maintained during gestation. Lead exposed groups had significant (at least p less than .01) maternal kidney and liver as well as fetal kidney lead content. Conceptus weight was significantly reduced in treatment groups. It was concluded that although significant amounts of lead crossed the placenta, as exemplified by fetal kidney values, no teratogenic response or reduction in fetal brain DNA content was produced in the rat.

A technique developed for isolated canine papillary muscle function in various plasmas; and utility ascertained with plasma containing myocardial depressant factors.

Isolated papillary muscles from young dogs were studied to determine the feasibility of utilizing plasma, without added foam depressants, as a test solution to support muscle function. Diluted canine plasma, undiluted canine plasma, plasma with adjusted protein content, monkey plasma, and plasma containing myocardial depressant factors (MDF) were evaluated. For each test, each papillary muscle served as its own control, with either Krebs-Henseleit solution or a control plasma to establish a basis for evaluation of the test plasma. A technique was implemented to oxygenate the muscle, in plasma, without foam production and to monitor pH continuously. These studies indicated that plasma (diluted, undiluted, or with protein adjustments to accommodate experimental procedural requirements) was a satisfactory test solution. Furthermore, plasma from species unrelated to the dog could be utilized. In utilizing plasma, it was essential to establish or adjust the pH and osmolarity for normal muscle function. These studies with plasma containing MDF indicated that MDF depressed canine papillary muscle function. The Vmax values and Po values were decreased and latency was increased. The effects of MDF in canine papillary muscle preparations were completely reversible in short-term studies (up 10 10 hours' duration).

Electrode catheterization for recording electrical activity of fasciculus atrioventricularis (bundle of His).

A method for locating an electrode catheter for observing electrical activities of the fasciculus atrioventricularis (bundle of His) without fluoroscopic guidance is described. A catheter which contains 4 electrodes and 2 internal capillary tubes was developed; the catheter has 2 distal orifices 18 mm apart. The position of the catheter tip is determined by observing blood pressure recordings from the two internal capillary tubes; when the tip is positioned properly, the proximal orifice will monitor atrial pressure and the distal orifice will detect ventricular pressure. This catheter was utilized on 14 mongrel dogs (19 to 27 kg). Recordings of atrioventricular bundle activities were successfully obtained from 12 of 14 dogs without any fluoroscopic guidances.

Plasma renin activity during controlled hemorrhage in the dog.

Purebred 1-year-old Beagles were made hypotensive by controlled hemorrhage (3 ml/kg-1/min-1). Femoral artery blood samples were collected every 5 minutes during controlled hemorrhage, until the blood pressure decreased to 30+/-5 mm Hg. Samples were then collected every 10 minutes for 40 minutes. Blood was not reinfused during the procedure. Carotid blood pressure, electrocardiogram lead II, and respiration rate were recorded. The results of the radioimmunoassay for renin activity indicated that arterial plasma renin activity formed a bimodal activity curve. Plasma renin activity did not reach any plateau, and it continued to increase throughout the experiment. Decreasing blood pressure and heart rate statistically showed a relationship with plasma renin activity (P less than 0.001). There was no interaction between pressure and heart rate; both were strong and independent. Female dogs had a higher plasma renin activity than did male dogs (P less than 0.001) throughout the experimental period. Plasma renin activity remained above resting activity throughout the development of hemorrhagic shock.