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To evaluate the effect of topical application of tetracaine on intraocular pressure (IOP) measurement by Tonopen in dogs.Six healthy male Epagneul Bretons (group 1) and six healthy male black Labrador Retrievers (group 2) were examined. IOP was measured in the right eye (OD) prior to (IOP1) and 1 minute following instillation of one drop of topical tetracaine (IOP 2), and the left eye (OS) (control) prior to (IOP 3) and 1 minute following instillation of one drop of isotonic saline solution (IOP 4). Measurements were performed on two occasions: at 8:00 AM and 3:00 PM.For both groups, IOP measurements were higher in the morning than in the afternoon. For group 1, IOP1 mean (SD), IOP2 mean (SD), IOP3 mean (SD) and IOP4 mean (SD) were 14.6 (2.2) mmHg, 11.3 (3.2) mmHg, 14.4 (2.2) mmHg and 13.5 (3.9) mmHg respectively, while in group 2, IOP1 mean (SD), IOP2 mean (SD), IOP3 mean (SD) and IOP4 mean (SD) were 14.2 (3.8) mmHg, 9.5 (3.7) mmHg, 13.5 (2.8) mmHg and 13.0 (3.8) mmHg respectively. For both groups at each time point, IOP 2 values were significantly lower (P0.007) than IOP 1 values, whereas IOP 3 and 4 values were not significantly different (P0.27).This study demonstrates that topical application of tetracaine significantly lowers IOP measured by Tonopen due to a possible interaction with melanin. The potential effect of topical anesthetics should be taken in consideration when performing applanation tonometry for clinical, pharmacological and toxicological studies.

SummaryCurrent therapy for hemophilia B requires large intravenous doses of factor IX (F.IX) given in the clinic or at home. Although home therapy is possible for many patients, it is often complicated by factors such as the lack of good venous access. Very little is known about extravascular routes for administering proteins like F.IX (57 kD) or other vitamin K-dependent procoagulant factors into the circulation. Questions about the absorption rate from extravascular administration as well as plasma recovery and bioavailability have arisen recently with the growing availibility of highly purified procoagulant proteins and increased interest in gene therapy of hemophilia B. Therefore, a group of studies were undertaken to determine the absorption rate, plasma recovery, and bioavailability of high purity, human plasma-derived F.IX concentrates administered via extravascular routes in hemophilia B dogs and in one human hemophilia B subject. Five hemophilia B dogs were given human F.IX via either a subcutaneous (SC), intramuscular (IM), intra- peritoneal (IP) or intravenous (IV) route. In a subsequent study, a single SC administration of human F.IX was compared to an identical IV dose of F.IX in the human hemophilia B subject. All extravascular routes of F.IX administration in both the canine and human gave lower levels of circulating plasma F.IX than the IV route, however all routes resulted in measurable F.IX activity. Of the extravascular routes, the IM injection in the canine resulted in a bioavailibility of 82.8%, while the SC injection resulted in a bioavailability of 63.5%. F.IX reached the plasma compartment by all extravascular routes used, confirming that F.IX can be absorbed extravascularly. The duration of measurable F.IX activity following extravascular administration is prolonged beyond that typically seen with IV administration. These data show that significant levels of F.IX may be obtained via SC injection in canine and ‘ human hemophilia B subjects and further highlight the potential of extravascular routes of administration for future experimental and clinical uses of F.IX and other procoagulant proteins.

Background— Phrenic nerve stimulation (PNS) is a common complication of cardiac resynchronization therapy when left ventricular (LV) pacing occurs via a coronary vein. The purpose of this study was to evaluate the effects of bipolar electrode spacing on PNS and LV pacing thresholds. Methods and Results— Electrophysiology catheters with standard (2 mm-5 mm-2 mm) or modified (1 mm-5 mm-1 mm) interelectrode spacing was, respectively, inserted in a posterior/lateral cardiac vein in a randomized order in 6 anesthetized dogs via jugular access. The phrenic nerve was dissected via a left minithoracotomy and repositioned over the vein as close as possible to one of the electrodes. The presence of PNS was verified (ie, PNS threshold P P >0.05). Compared with a standard bipolar electrode spacing of 20 mm for LV leads, 1 and 2 mm bipolar electrode spacing resulted in a PNS threshold increase of 5.5±2.2 V ( P =0.003) and 2.8±1.7 V ( P P Conclusions— This study suggests that reducing LV bipolar electrode spacing from the standard 20 mm to 1 or 2 mm may significantly increase the PNS threshold without compromising LV pacing thresholds.

Quantitative analysis has demonstrated five temporal stages of activation during the first 10 min of ventricular fibrillation (VF) in dogs. To determine whether these stages exist in another species, we applied the same analysis to the first 10 min of VF recorded in vivo from two 504-electrode arrays, one each on left anterior and posterior ventricular epicardium in six anesthetized pigs. The following descriptors were continuously quantified: 1) number of wavefronts, 2) wavefront fractionations, 3) wavefront collisions, 4) repeatability, 5) multiplicity index, 6) wavefront conduction velocity, 7) activation rate, 8) mean area activated by the wavefronts, 9) negative peak rate of voltage change, 10) incidence of breakthrough/foci, 11) incidence of block, and 12) incidence of reentry. Cluster analysis of these descriptors divided VF into four stages ( stages i-iv). The values of most descriptors increased during stage i (1–22 s after VF induction), changed quickly to values indicating greater organization during stage ii (23–39 s), decreased steadily during stage iii (40–187 s), and remained relatively unchanged during stage iv (188–600 s). The epicardium still activated during stage iv instead of becoming silent as in dogs. In conclusion, during the first 10 min, VF activation can be divided into four stages in pigs instead of five stages as in dogs. Following a 16-s period during the first minute of VF when activation became more organized, all parameters exhibited progressive decreased organization. Further studies are warranted to determine whether these changes, particularly the increased organization of stage ii, have clinical consequences, such as alteration in defibrillation efficacy.

Influence of 4,5-dihydro-6-[1-[2-hydroxy-2-(4-cyanophenyl)ethyl]- 1,2,5,6,-tetrahydropyrido-4-yl]pyridazin-3(2H)-one (SCH00013) and vesnarinone (CAS 81840-15-5) on the arrhythmia experimentally induced by three different methods was investigated in dogs. In digitalis-induced arrhythmia, SCH00013 (3 mg/kg i.v.) showed a tendency to improve the arrhythmia with a decrease in the arrhythmic ratio and an increase in the conducted beats (CB), though these changes did not reach a significant level; it decreased significantly the blood pressure (BP) with no change in the total heart rate (THR) and atrial rate (AR). Vesnarinone (3 mg/kg i.v.) did not affect these parameters except for BP that was decreased significantly. In two-stage coronary ligation-induced arrhythmia, SCH00013 (1 and 3 mg/kg i.v.) did not change the arrhythmic ratio, CB, AR and BP, while the THR being slightly decreased; the arrhythmic ratio showed a tendency to decrease with SCH00013 when examined at 24 h after coronary ligation. Vesnarinone (3 mg/kg i.v.) did not affect these parameters at 24 and 48 h after ligation. In epinephrine (adrenaline)-induced arrhythmia, both SCH00013 and vesnarinone showed exacerbation of arrhythmia. SCH00013 at 1 mg/kg i.v. did not elicit ventricular fibrillation (VF) in five dogs examined, but at 3 mg/kg i.v. it elicited VF in two of three dogs. Vesnarinone at 1 mg/kg i.v. induced VF in all of three dogs examined. Incidence of VF induced by optical isomers of SCH00013 was not significantly different from each other: both isomers elicited VF in two of six dogs at 1 mg/kg i.v. and at 3 mg/kg i.v. each of them induced VF in two dogs examined. The present results indicate that SCH00013 is a cardiotonic agent that is equivalent to or less arrhythmogenic than vesnarinone in animal models of arrhythmia, such as adrenaline- and digitalis-induced arrhythmia and the two-stage coronary ligation-induced arrhythmia. Optical isomers of SCH00013 were essentially equieffective in eliciting exacerbation of adrenaline-induced arrhythmia in the dog.

The insulin-sensitizing agent troglitazone ((+/-)-5-[4-(6-hydroxy-2,5,7,8-tetramethylchroman-2-ylmethoxy++ +)benzyl]-2,4- thiazolidinedione, CAS 97322-87-7), has been reported to reduce vascular tone in the rat tail artery. This study was designed to examine whether troglitazone could relax the precontracted canine coronary artery in vitro. Isometric tension of coronary arterial rings was measured, and analysis by Mann-Whitney U-test revealed that significant relaxation was induced even with 1 mumol/l troglitazone. This relaxation was not affected by blocking of NO synthase, cyclooxygenase, the large and intermediate conductance Ca(2+)-activated K+ channel, the ATP-sensitive K+ channel, or the Na(+)-K+ pump. The exact mechanism of the reduction of vascular tone by troglitazone remains unclear. However, the finding that troglitazone acts as a vasorelaxant in the coronary artery may be clinically useful, since troglitazone is mainly used for type II diabetic patients with insulin resistance, which is associated with increased risk for coronary artery disease.

The general pharmacological properties of 2-amino-9-(3-acetoxymethyl-4-isopropoxycar-bonyloxybut-1-yl) purine (CAS 247081-81-8, SK 1899), a new potential antiviral agent, were investigated in mice, rats, guinea pigs, rabbits, and dogs. The oral administration of 50, 150, and 500 mg/kg of SK 1899 had no effects on the central nervous system except that it slightly increased the spontaneous locomotor activity in mice at a dose of 500 mg/kg. SK 1899 did not disturb either the spontaneous motility or contractor-induced contraction of the isolated organs such as guinea pig ileum, rat uterus, guinea pig vas deferens, and guinea pig trachea at concentrations up to 10 –4 mol/l. It slightly increased the contractile force in the isolated guinea pig atrium at a concentration of 10 –4 mol/1. Following intravenous infusion of 5, 15, and 50 mg/kg of SK 1899 to anesthetized dogs, it did not change the mean arterial pressure, heart rate, left ventricular systolic pressure (LVSP), and respiratory rate, while it slightly increased the left ventricular positive dP/dt max (LV + dP/dt max ) at a dose of 50 mg/kg. SK 1899 did not induce any significant changes in the intestinal charcoal meal transit in mice, basal gastric juice secretion in rats, and renal function in rats. It did not affect the blood coagulation system and phenolsulfonphthalein secretion in rats. These findings suggest that SK 1899 has a very low potential to induce any adverse pharmacological effects at the doses showing antiviral activity.

An in vivo preclinical study has been made of the oral absorption of morphine (CAS 57-27-2) from a new sustained release formulation (morphine-Eudragit® L complex, MEC), which had shown good sustained release properties in in vitro dissolution studies. The absorption of morphine from capsules filled with morphine hydrochloride trihydrate (MHT) or MEC was compared in fasted and fed dogs. Mean plasma morphine concentrations obtained after administration of MHT and MEC to fasted dogs were similar, and no statistically significant differences were found in the pharmacokinetic parameters of morphine (C max , T max and area under the plasma morphine concentration versus time curve from time zero to the last time with a detectable concentration of morphine). When MHT and MEC were administered to fed animals, mean plasma morphine concentrations were again similar for both formulations, without statistically significant differences in the pharmacokinetic parameters of morphine. These results contrast with those obtained in vitro , and indicate the limited usefulness of in vitro assays for this kind of sustained release formulations in which pH and ionic strength are important factors for drug release from the polymeric structure. The plasma morphine concentrations obtained in fed dogs were generally lower than in fasted dogs, though they were detectable for a longer time, until 10 h after dosing, in contrast to up to 6 h in fasted dogs. It is postulated that the apparently prolonged absorption of morphine in fed dogs may be due to the enterohepatic recycling of the drug (excreted in bile as glucuronide, hydrolysed back to the parent compound in the intestine, and then reabsorbed) as a consequence of gallbladder emptying induced by food.

Aim NN1731 is a recombinant activated factor VII (rFVIIa) analogue with enhanced activity. The objective of the present study was to evaluate the clearance mechanisms of rFVIIa and NN1731 after intravenous administration to Beagle dogs. Methods The study was performed in Beagle dogs administered with a single dose of 5.4 nmol/kg rFVIIa or NN1731 intravenously. Plasma samples collected up to 12-h post-administration were analysed using three different assays to determine FVIIa clot activity (FVIIa:C), total FVIIa antigen, and levels of FVIIa–antithrombin (AT) complexes. Pharmacokinetic parameters were determined by use of standard non-compartmental and non-linear mixed effects methods. Results For both compounds, complex formation with AT accounted for the observed difference between the activity and the antigen curves and constituted 60–70% of the total clearance. The clearance of rFVIIa and NN1731 was estimated to be 73 and 214 mL/h/kg, respectively, accordingly, AT complex formation occurred around three times faster for NN1731. The difference in activity observed in the initial phase, resulting in distribution half-lives of 0.71 and 0.22 h for rFVIIa and NN1731, was mainly caused by the 3-fold difference in clearance. The terminal half-life of rFVIIa and NN1731 was estimated to be 2.1 and 2.5 h, respectively. The non-compartmental analysis resulted in almost identical parameters. Conclusion The present study demonstrates that the difference between the activity and the antigen profiles of rFVIIa and NN1731 in Beagle dogs is the result of complex formation with AT which constitutes a major pathway for the clearance of rFVIIa activity.

Amylase-resistant, periodic acid-Schiff (PAS)–positive inclusions were identified in the skeletal muscle of four of twenty-four purpose-bred beagle dogs from a routine toxicology study. Affected myofibers contained amorphous material filling up to 20% of the sarcoplasm that stained lightly basophilic with hematoxylin and eosin and was strongly PAS–positive with amylase resistance. Transmission electron micrographic examination of the inclusions revealed granular, non-membrane–bound, electron-dense material, consistent with polysaccharide. Although skeletal muscle inclusions with similar features have been reported in dogs in conjunction with systemic metabolic disorders and less often in muscle adjacent to nonmyogenic sarcomas, all four of these dogs lacked clinical or pathological findings diagnostic of a concurrent systemic metabolic or localized skeletal muscle disorder. Furthermore, these skeletal muscle inclusions were present in both vehicle- and test article–treated dogs and were considered an incidental finding that may occur spontaneously in clinically normal beagle dogs; as such, their presence in drug-treated animals should be interpreted with caution.

Prelamination is sometimes required for reconstruction of specialized organs. The purpose of this study was to create a chimeric flap model in dogs for prelamination. The anatomy of the deep and superficial caudal epigastric vessels, which supply the rectus abdominis muscle and superficial ventral fascia, respectively, were measured in 14 dogs. Their lengths were ~30 mm with the arteries averaging 1.4 mm and veins, 2.5 mm in diameter. These vessels shared a 10-mm-long common trunk with the artery averaging 2 mm and the vein, 3.2 mm in diameter. Chimeric flaps were successfully raised in six dogs based on the common vascular trunk, and they remained well perfused 3 weeks after prelamination. The perfusion territory of the abdominal skin measured 25 cm long and 10 cm wide with methylene blue injection. This study suggests that the caudal epigastric system can be a suitable chimeric flap model in dogs.

The safety and consequences of fine- (FNA) and large-needle aspiration (LNA) to the testicular parenchyma and its normal function have not been thoroughly established. This study was performed to accurately assess, by serial clinical, in vivo ultrasonographic and seminological examinations, the type and extent of the effect of FNA or LNA on canine testes. Eighteen sexually mature, 1-2 years old, healthy laboratory Beagles were used. One of their testes was aspirated using a 23-G butterfly needle (FN) and the other using a 19-G butterfly needle (LN). Two dogs at a time were orchiectomized 10, 60 min, 2, 14, 29, 63, 76, 90 or 180 days post-aspiration. Five and 2 days and 1 h before aspiration (in all dogs), immediately post-aspiration, and 1, 2, 4, 7, 9, 14, 19, 29, 35, 43, 49, 56, 63, 70, 76, 90, 111, 132 and 180 days post-aspiration (in the remaining intact dogs), evaluation of scrotal surface temperature over each testis, evaluation of scrotum-testis volume by electronic sliding callipers, ultrasonographic evaluation of testicular volume and texture and clinical and semen examination were performed. Following FNA and LNA, the clinical and ultrasonographic appearance of the testis were normal. Sperm production nearly always remained unchanged, with the exception of a slight decrease in spermatozoal motility 2-14 days post-aspiration. However, even then, with the exception of six samples, spermatozoal motility was above normal values. Within the parameters of this experiment, testicular FNA and LNA have no ill effect on sperm production or clinical and ultrasonographic appearance of the canine testis, and therefore, both FNA and LNA should be considered safe.

The exercise pressor reflex is due to activation of thin fiber afferents within contracting muscle. These afferents are in part stimulated by ATP activation of purinergic 2X (P2X) receptors during contraction. Which of the P2X receptors contribute to the reflex is unknown; however, P2X2/3 and P2X3 receptor subtypes are good candidates because they are located on thin fiber afferents and are involved in sensory neurotransmission. To determine if P2X2/3 and P2X3 receptors evoke the metabolic component of the exercise pressor reflex, we examined the effect of two P2X2/3 and P2X3 antagonists, A-317491 (10 mg/kg) and RO-3 (10 mg/kg), on the pressor response to injections of α,β-methylene ATP (α,β-MeATP; 50 μg/kg), freely perfused static contraction, contraction of the triceps surae muscles while the circulation was occluded, and postcontraction circulatory occlusion in decerebrate cats. We found that the antagonists reduced the pressor response to α,β-MeATP injection (before Δ 20 ± 3 mmHg; drug Δ 11 ± 3 mmHg; P < 0.05), suggesting the antagonists were effective in blocking P2X2/3 and P2X3 receptors. P2X2/3 and P2X3 receptor blockade reduced the pressor response to freely perfused contraction (before Δ 33 ± 5 mmHg; drug Δ 15 ± 5 mmHg; P < 0.05), contraction with the circulation occluded (before Δ 52 ± 7 mmHg; drug Δ 20 ± 4 mmHg; P < 0.05), and during postcontraction circulatory occlusion (before Δ 15 ± 1 mmHg; drug Δ 5 ± 1 mmHg; P < 0.05). Our findings suggest that P2X2/3 and P2X3 receptors contribute to the metabolic component of the exercise pressor reflex in decerebrate cats.

Objective The liver is susceptible to ischemia-reperfusion (IR) injury during inflow occlusion for hepatectomy. There is no effective pharmacologic agent available to prevent the release of high-mobility-group box 1 (HMGB1) or to ameliorate IR injury. This pilot study sought to develop a model in beagle dogs for the purpose of testing the efficacy of a necrosis modulator, necrox-7, to prevent hepatic IR injury in beagle dogs. Methods Six male beagle dogs were randomly assigned to the control group (group A; n = 3) or the treatment group (group B; n = 3). Under general anesthesia, group B received intravenous infusion of necrox-7 (13 mg/kg over 20 minutes) followed by 60 minutes of left hepatic inflow occlusion and 60 minutes of reperfusion. Both groups were tested for serum biochemicals, hematology values, liver biopsies, and plasma HMGB1 levels over a 48-hour period. Results The maximum alanine transferase (ALT), aspartate transferase (AST), and lactate dehydrogenase (LDH) levels among group A versus group B were: ALT 868.3 ± 337.4 IU/L vs 274.3 ± 72.6 IU/L ( P = .041); AST 1,024.7 ± 246.5 IU/L vs 505.3 ± 66.7 IU/L ( P = .024); and LDH 962.7 ± 226.2 IU/L vs 552.7 ± 62.4 IU/L ( P = .039). Liver biopsy demonstrated marked necrosis and inflammatory infiltrates in group A, whereas group B showed little evidence of IR injury. The plasma HMGB1 concentration was significantly lower among group B versus A. Conclusion This pilot study developed a hepatic IR injury model, demonstrating that necrox-7 reduced hepatic necrosis secondary to IR injury in a large animal setting.

GnRH (gonadotrophin releasing hormone) is a key hormone of reproductive function in mammals; agonist forms have been largely developed, and data concerning their use in small animal reproduction are now abundant. GnRH agonists act by a two-step mechanism. First, their agonist properties on the pituitary will cause marked LH (luteinizing hormone) and FSH (follicle-stimulating hormone) secretion into the bloodstream, accompanied by an increase in the concentrations of sex steroid hormones. Then, in case of constant administration, GnRH agonists will lead to pituitary desensitization, and FSH and LH levels will collapse. These two effects have been widely documented, and these compounds have many potential benefits in a clinical context, capitalizing both on their stimulating and sterilizing effects.

Background— The left ventricular (LV) dilatation of isolated mitral regurgitation (MR) is associated with an increase in chymase and a decrease in interstitial collagen and extracellular matrix. In addition to profibrotic effects, chymase has significant antifibrotic actions because it activates matrix metalloproteinases and kallikrein and degrades fibronectin. Thus, we hypothesize that chymase inhibitor (CI) will attenuate extracellular matrix loss and LV remodeling in MR. Methods and Results— We studied dogs with 4 months of untreated MR (MR; n=9) or MR treated with CI (MR+CI; n=8). Cine MRI demonstrated a >40% increase in LV end-diastolic volume in both groups, consistent with a failure of CI to improve a 25% decrease in interstitial collagen in MR. However, LV cardiomyocyte fractional shortening was decreased in MR versus normal dogs (3.71±0.24% versus 4.81±0.31%; P Conclusions— These results suggest that chymase disrupts cell surface–fibronectin connections and FAK phosphorylation that can adversely affect cardiomyocyte myofibrillar structure and function. The greater effect of CI on epicardial versus endocardial titin and noncollagen cell surface proteins may be responsible for the increase in torsion angle in chronic MR.

The objective of the present study was to systematically evaluate the impact of methodology on thromboelastometry with canine whole blood. Thromboelastometry was performed on citrated blood using a variety of combinations of clotting activators [ex-tem (tissue factor or TF), in-tem (ellagic acid), diluted TF from Innovin, or Ca (recalcification only)] and storage times. Thromboelastometry was also performed using diluted TF from Innovin on blood collected into a contact inhibitor. Ex-vivo contact activation was compared between canine and human blood. Clotting activator had a marked impact on coagulation time, a minor impact on alpha angle, and no impact on clot formation time or maximum clot firmness. When ex-tem or in-tem was the clotting activator, sample storage up to 30 min did not affect results. With diluted TF from Innovin or Ca, sample storage was associated with the development of increased coagulability (as indicated by shorter coagulation time and clot formation time and higher alpha angle) due to ex-vivo contact activation. Canine blood underwent markedly more ex-vivo contact activation than did human blood. Canine blood undergoes significant ex-vivo contact activation during and after collection, which influences thromboelastometry results when a weak clotting activator (such as low TF or recalcification) is used. Thromboelastometry with a strong activator (such as ex-tem or in-tem) is less influenced by ex-vivo changes, and, therefore, likely to be more reflective of in-vivo hemostatic capabilities and to provide consistently interpretable and comparable results.

Background: Tendon surface modification with a synthetic biopolymer, carbodiimide-derivatized hyaluronic acid and gelatin with the addition of lubricin (CHL), has been shown to reduce gliding resistance after tendon repair in an in vitro model. The purpose of the study was to investigate whether CHL would reduce adhesion formation and improve digital function after flexor tendon repair in a canine model in vivo. Methods: Sixty dogs were randomly assigned to either a biopolymer-treated group (n = 30) or an untreated control group (n = 30). The second and fifth flexor digitorum profundus tendons from each dog were lacerated fully at the zone-II area and then repaired. Passive synergistic motion therapy was started on the fifth postoperative day and continued until the dogs were killed on day 10, day 21, or day 42. The repaired tendons were evaluated for adhesions, normalized work of flexion, gliding resistance, repair strength, stiffness, and histological characteristics. Results: The normalized work of flexion of the repaired tendons treated with CHL was significantly lower than that of the non-CHL-treated repaired tendons at all time points (p < 0.05), and the prevalence of severe adhesions was also significantly decreased in the CHL-treated tendons at day 42 (p < 0.05). However, the repair failure strength and stiffness of the CHL-treated group were also significantly reduced compared with those of the control group at days 21 and 42 (p < 0.05) and the rate of tendon rupture was significantly higher in the treated group than in the control group at day 42 (p < 0.05). Conclusions: Treatment with the lubricin-containing gel CHL appears to be an effective means of decreasing postoperative flexor tendon adhesions, but it is also associated with some impairment of tendon healing. Future studies will be necessary to determine if the positive effects of CHL on adhesion formation can be maintained while reducing its adverse effect on the structural integrity of the repaired tendon.

Molecular weight (MW) is known as an important factor of biliary excretion in rats, guinea pigs, rabbits and humans. The objective of this study was to evaluate the relationship between the biliary excretion and MW of drugs in dogs. Data on the percentage of dose excreted into bile as parent drug (PD(b)) in dogs were collected from the literature for 134 compounds. Receiver operating characteristic (ROC) curve analysis was utilized to determine whether a MW threshold exists for PD(b). A MW threshold of 375-400 Da was established for anions in dogs, which is similar with the cutoff value observed in rats (400 Da) but lower than the one in humans (475 Da). No MW threshold was found for cations or cations/neutral compounds. A molecular volume threshold of 300A(3) was also determined for anions in dogs, which corresponds to a MW of 394 Da. In conclusion, our analysis suggested the presence of a MW cutoff for anions in dogs, which may be related with the molecular size of a compound. This represents the first report of the influence of MW or molecular volume as a determinant of biliary excretion for a structurally diverse set of compounds in dogs.

Background CW002 is a neuromuscular blocking drug that is inactivated by endogenous L-cysteine. This study determined the exogenous L-cysteine dose-response relationship for CW002 reversal along with acute cardiovascular effects and organ toxicity in dogs. Methods Six dogs were each studied four times during isoflurane-nitrous oxide anesthesia and recording of muscle twitch, arterial pressure, and heart rate. CW002 (0.08 mg/kg or 9 x ED95) was injected, and the time to spontaneous muscle recovery was determined. CW002 was then administered again followed 1 min later by 10, 20, 50, or 100 mg/kg L-cysteine (1 dose/experiment). After twitch recovery, CW002 was given a third time to determine whether residual L-cysteine influenced duration. Preliminary toxicology was performed in an additional group of dogs that received CW002 followed by vehicle (n = 8) or 200 mg/kg L-cysteine (n = 8). Animals were awakened and observed for 2 or 14 days before sacrificing and anatomic, biochemical, and histopathologic analyses. Results L-cysteine at all doses accelerated recovery from CW002, with both 50 and 100 mg/kg decreasing median duration from more than 70 min to less than 5 min. After reversal, duration of a subsequent CW002 dose was also decreased in a dose-dependent manner. Over the studied dose range, L-cysteine had less than 10% effect on blood pressure and heart rate. Animals receiving a single 200-mg/kg dose of L-cysteine showed no clinical, anatomic, biochemical, or histologic evidence of organ toxicity. Conclusion The optimal L-cysteine dose for rapidly reversing the neuromuscular blockade produced by a large dose of CW002 in dogs is approximately 50 mg/kg, which has no concomitant hemodynamic effect. A dose of 200 mg/kg had no evident organ toxicity.

Our aim was to explore the character of distracted bone in irradiated canine mandibles and the electrophysiological changes in the irradiated inferior alveolar nerve (IAN). Twelve Chinese dogs were studied, 10 of which were given unilateral irradiation of (60)Co in the mandible with a total dose of 22.8Gy in four 5.7Gy fractions (biologically equivalent to 50Gy/25 fractions) (experimental group). The other two dogs were not irradiated and served as controls. All had a bilateral corticotomy 3 months after irradiation. After a 1-week latency period distraction of the mandible was activated at a rate of 0.5mm twice daily for 10 days, followed by a consolidation phase of 8 weeks. New bone was assessed by radiographic, histological, and single-photon electron computed tomographic (SPECT) analysis. The IAN was analysed electrophysiologically. One dog in the experimental group was excluded from the study with anaesthetic problems. After 8 weeks of consolidation there was no difference between the percentage area of new bone in the two groups. New bone was more mature and organised in the control group than in the experimental group. SPECT analysis showed that there was active osteogenic activity in dogs in the experimental group. The action potential of the IAN showed corresponding changes during the irradiation and distraction processes. We conclude that distraction osteogenesis is feasible in previously irradiated canine mandibles and IAN.

The aim of this study was to investigate the feasibility of employing S-carboxymethyl-L-cysteine as a treatment of chronic obstructive pulmonary disease in dogs. To this end the pharmacokinetic parameters of orally administered S-carboxymethyl-L-cysteine were determined in the dog, cow and sheep. Six healthy beagle dogs, six endogenous Greek sheep and four Holstein Fresian calves were orally dosed with 10 mg/kg body weight of S-carboxymethyl-L-cysteine. No significant differences in T(max) and T(1/2) were reported between the species. However, significantly higher AUC((0-last)), 21.56+/-6.67 microg h ml(-1) and AUC((0-infinity)), 21.63+/-6.68 microg h ml(-1) were seen in the dogs compared to the sheep and calves. The calculated V(D) was significantly higher in the sheep (10.4+/-2.7 L kg(-1)) and the calves (3.8+/-0.7 L kg(-1)) compared to the dogs (1.0+/-0.6 L kg(-1)). The rank order of increasing C(L) was sheep (3.4+/-2.7 L h(-1)kg(-1))>calves (2.7+/-0.4 L h(-1) kg(-1))>dogs (0.5+/-0.2 L h(-1)kg(-1)). The result for the dogs was significantly lower that the calculated C(L) for the sheep and calves. All these results indicate that the oral administration of S-carboxymethyl-L-cysteine may be useful during the therapeutic management of chronic obstructive pulmonary disease in dogs.

This study assessed the effect of recombinant fibroblast growth factor (FGF-2) on reimplanted teeth.Premolars were reimplanted in dogs. Roots on the left side of the mandible were treated with FGF-2, whereas roots on right side served as controls. At 2, 4, or 8 weeks after treatment, animals were humanely killed and specimens collected and processed for histopathologic examination.After 2 weeks, proliferating cell nuclear antigen-positive cell ratios in the experimental group were significantly greater than those in the control group (P.01). After 4 and 8 weeks, the experimental group showed formation of new cementum. Moreover, incidence of replacement resorption in the experimental group was significantly lower than that in the control group (P.01).The results indicate that FGF-2 promotes formation of new periodontal ligament and prevents ankylosis and root resorption following reimplantation of teeth.

PK/PD analysis was performed after percutaneous administration of insulin self-dissolving micropiles (SDMP) where chondroitin sulfate was used as a base. PK/PD model composed of 1-compartment open model with first-order absorption process and sigmoidal Emax model was used to simulate the time courses of both plasma insulin and glucose levels after percutaneous administration to dogs, 2.0 IU/dog. The Akaike’s information criterion (AIC) were 249.33 ± 13.00, though AIC was 259.55 ± 11.95 for the 2-compartment open model that was directly linked to sigmoidal E max model. The mean first-order absorption and elimination rate constants of insulin with SDMP were 7.59 ± 3.93 h − 1 (k 01 ) and 1.31 ± 0.66 h − 1 (k 10 ). For subcutaneous (sc) injection of insulin solution, k 01 and k 10 were 9.72 ± 5.67 h − 1 and 1.60 ± 0.45 h − 1 . PD parameter values from insulin SDMP were 148.5 ± 71.9 pg/mL for EC50 and 3.11 ± 1.41 for γ. These results suggested that insulin SDMP has almost the same PK characteristics as sc injection preparation.

This study analyzes the parathyroid function in four dogs before and after 2 years of a low-calcium, high-sodium, vitamin D-deficient diet and the involution of the same function following (1) correction of dietary calcium deficiency and administration of IV 1,25-(OH)2D (0.25 μg twice per day) during 1 month, (2) after an additional month of normal dog chow supplemented with oral vitamin D (25 μg per day), and, finally, (3) after 5 and 17 months of a diet with normal levels of calcium and vitamin D. The parathyroid function was evaluated through IV infusion of CaCl2 and Na2 EDTA with measurement of intact (I) and carboxyl-terminal (C) immunoreactive parathyroid hormone (iPTH). The C-iPTH/I-iPTH ratio was calculated to assess the modulation of molecular forms of iPTH induced by the various treatments. The 2 years of calcium and vitamin D deprivation lowered ionized calcium (1.23 ± 0.04, p < 0.05) and 25-OHD (4.02 ± 2.06 nM, p < 0.005) and tended to decrease 1,25-(OH)2D (80.8 ± 8.6 pM); it increased basal I- and C-iPTH levels approximately eightfold (I-iPTH, 40.2 ± 20.7, p < 0.05; C-iPTH, 185.4 ± 94.9, p < 0.05) and stimulated I-iPTH (60.2 ± 23.0 pM, p < 0.05) and C-iPTH (239.6 ± 80.7 pM, p < 0.05) fivefold. A greater rise in nonsuppressible I-iPTH levels than in C-iPTH levels led to a decreased C-iPTH/I-iPTH ratio in hypercalcemia (12.5 ± 2.8 versus 27.8 ± 6.05 pM, p < 0.005). Dietary deprivation also decreased the I-iPTH calcium stimulation set point (1.25 ± 0.05 pM, p < 0.05). Treatment with dietary calcium and IV 1,25-(OH)2D normalized ionized calcium (1.34 ± 0.02 mM) and basal I-iPTH level (6.09 ± 5.48 pM) more than basal C-iPTH (69.8 ± 48.8 pM), causing the C-iPTH/I-iPTH ratio to increase from 4.80 ± 0.81 to 14.8 ± 6.1 (p < 0.05). Stimulated I-iPTH decreased (46.1 ± 15.3 pM, p < 0.05), but stimulated C-iPTH secretion did not change (282.4 ± 90.9 pM). A greater fall in the nonsuppressible I-iPTH level as opposed to C-iPTH level led the C-iPTH/I-iPTH ratio to increase back to a normal value (23.8 ± 1.40). The I-iPTH calcium stimulation set point remained low at 1.28 mM. Further consecutive treatment modalities over 18 months did not cause additional significant change in basal, stimulated, or nonsuppressible C-iPTH levels, but the same I-iPTH levels tended to decrease further. This explained why all three C-iPTH/I-iPTH ratios tended either to increase (basal) or to increase significantly (stimulated and nonsuppressible, p < 0.05). The I-iPTH calcium stimulation set point again remained low. This study reveals that animals with hyperplastic parathyroid glands can control their I-iPTH level by maintaining a lower set point of I-iPTH stimulation by calcium and by changing their secretion profile with respect to carboxyl-terminal fragments of the PTH molecule.

Cyclic mechanical loading sufficient to engender strains of physiologic magnitude applied to recently excised canine cancellous bone cores in vitro increased the release of prostaglandin E (PGE) and prostacyclin (PGI2, measured as its breakdown product 6-keto-PGF1 alpha), during a 15 minute loading period in which PG levels were measured in perfusing medium at 5 minute intervals. Peak production occurred in the 0-5 minute sample. Mean levels preload compared to during load were PGE, 2.66 and 3.67 ng/ml (p less than 0.002); and 6-keto-PGF1 alpha, 543 and 868 pg/ml (p less than 0.007). The elevated levels then declined to preload levels during the loading period. However, the 5-10 minute but not the 10-15 minute samples still contained levels greater than preload values. A second 15 minute period of load, 1 h following the end of the first, produced smaller increases in the levels of release that were statistically significant only for the first 0-5 minute sample during load (preload compared to load mean values, PGE, 1.09-1.66 ng/ml, p less than 0.02; 6-keto-PGF1 alpha, 401-558 pg/ml, p less than 0.04). Immunolocalization revealed PGE and 6-keto-PGF1 alpha in lining cells and 6-keto-PGF1 alpha but not PGE in osteocytes. Addition to the medium of 1 microM PGE2, approximating the concentration produced by loading, had no significant effect on the specific activity of the extractable RNA fraction labeled with [3H]uridine, whereas 1 microM PGI2 produced an increase similar to that seen previously with loading.(ABSTRACT TRUNCATED AT 250 WORDS)

To investigate the dose-dependent effects of risedronate on cancellous bone remodeling, adult female beagle dogs were treated with either placebo, 0.1, 0.5, or 2.5 mg/kg/day of risedronate orally in an intermittent cyclic regimen (7 days on 21 days off), repeated three times. Iliac cancellous bone samples were subjected to histomorphometric analysis and three-dimensional (3-D) kinetic reconstruction of the remodeling site was performed. In the 0.1 mg/kg dose group, resorption and activation indices were no different from the placebo group. However, wall thickness was increased resulting in a positive bone balance at the level of the remodeling unit. In the 0.5 and 2.5 mg/kg dose groups, a dose-dependent reduction in activation frequency and tissue level bone formation was observed. Resorption rates were also significantly decreased, 60% and 80% for the 0.5- and 2.5-mg/kg groups, respectively. An approximate 25% reduction in final erosion depth was noted in both these groups. Analyses of the growth curves of the bone packet confirmed that the kinetics of the growth of a completed packet were different in the 0.5- and 2.5-mg/kg dose groups compared with placebo. These changes were associated with a significant increase in the final wall thickness in both groups indicating no net impairment of osteoblast function. These increases in wall thickness in combination with the reductions in final erosion depth in the 0.5 and 2.5 mg/kg groups resulted in a significant dose-dependent positive bone balance. This pharmacological profile suggests that risedronate may be of therapeutic utility in the treatment of metabolic bone diseases where reductions in activation frequency and resorptive cell activity at the level of the remodeling unit are a therapeutic goal.

Young canines born at sea level (SL) and raised for 5 mo at high altitude (HA, 3,800 m), followed by return to SL before somatic maturation, showed enhanced alveolar gas exchange and diffusing capacity at rest and exercise that persisted into adulthood (McDonough P, Dane DM, Hsia CC, Yilmaz C, Johnson RL Jr. J Appl Physiol 100: 474–81, 2006; Hsia CCW, Johnson RL Jr, McDonough P, Dane DM, Hurst MD, Fehmel JL, Wagner HE, Wagner PD. J Appl Physiol 102: 1448–55, 2007). To examine the associated structural response, we quantified lung ultrastructure in male foxhounds raised at 3,800 m HA or their littermates raised at SL ( n = 6 each) from 2.5 to 7.5 mo of age. Three years following return to SL, lungs were fixed for morphometric analysis. In HA-exposed animals compared with SL controls, lung volume at a given inflation pressure was higher with enlargement of alveolar ducts and sacs without significant differences in the volumes of alveolar cell components, septal tissue, or in alveolar-capillary surface areas. There was a shift toward a significantly lower harmonic mean thickness of the blood-gas diffusion barrier in HA-raised animals. As a control organ, muscle capillary length density of costal diaphragm was significantly higher in HA-raised animals, indicating parallel adaptation in oxygen transport organs. We conclude that, in actively growing animals, 5 mo of HA exposure that was discontinued before somatic maturation induced acinar remodeling that increased lung compliance and reduced the resistance of blood-gas diffusion barrier to diffusion that persisted into adulthood, but without permanent enhancement of alveolar tissue growth.

The anti-cancer chemotherapeutic agent cisplatin induces an acute (∼24 h) and delayed (∼24–72 h+) emetic response in humans; whereas the mechanism mediating the acute phase has been characterised, the delayed phase is relatively poorly understood. We have used nerve lesions (abdominal vagus, VX; greater splanchnic nerve, GSNX) and area postrema ablation (APX) in the ferret model of cisplatin (5 mg/kg, i.p.) delayed emesis and demonstrated that VX and VX + GSNX did not significantly modify the delayed emetic response (24–72 h), which consisted of 276.0 ± 62.8 retches + vomits (R + V) in sham-operated ferrets and 167.2 ± 34.0 R + V and 214.8 ± 40.2 R + V, in the VX and VX + GSNX groups, respectively. APX virtually abolished the delayed phase of emesis and sham-operated ferrets had 93.0 ± 22.9 R + V whilst only 6.0 ± 3.6 R + V ( p = 0.009) were observed in APX animals. These data suggest that, in contrast to the acute emetic response triggered by cisplatin, the delayed phase does not rely on abdominal visceral afferents but is mediated via the area postrema.

The mechanism for the initiation of giant migrating contractions (GMCs) associated with defecation is not well known. The aim of this study was to describe the characteristics of special propulsive contractions (SPCs), such as GMCs, during evacuation in four experimental dog models, with emphasis on denervation. Twenty healthy dogs were used in this study, and they were divided into four groups, i.e., control (underwent force transducer implantation alone), denervation (underwent transection of the descending nerve fibers along the caudal mesenteric artery (CMA)), transection (underwent transection of the rectum, which corresponds to transection of the enteric nerve fibers), and denervation-plus-transection (underwent transection of the descending nerve fibers along the CMA and transection of the rectum). Colonic contractile activities were continuously recorded on a computer. Five force transducers were implanted at the serosal surfaces of the colon (C1-R). The consistency of dog feces was checked daily. The parameters of rectal relaxation (RR), defecation characteristics, and SPCs, such as motility index (MI), duration, and frequency, were measured. In the control and denervation groups, GMCs were observed with evacuation, and RR occurred synchronously with the initiation of GMCs. On the other hand, in the transection and denervation-plus-transection groups, strong force contractions without RR occurred only during evacuation. The MI and duration of the transection and denervation-plus-transection groups were higher than those of other groups (p

We tested the effect of dose of GnRH superagonist on pituitary and testicular function in a study with four groups of four male dogs. The Controls received blank implants and the other three groups received implants containing 3, 6 or 12 mg deslorelin (D-Trp6-Pro9-des-Gly10-GnRH ethylamide). In all deslorelin-treated groups, there was initially an acute increase in plasma concentrations of LH and testosterone, followed by declines such that both hormones became undetectable after approximately 12 days. There was a dose-response in some of these early aspects of the hormone profiles. With respect to long-term effects of treatment, the 12-mg dose had significantly greater effects than the smaller doses for the duration of minimum testicular volume [366 +/- 77, mean +/- SEM (3 mg), 472 +/- 74 (6 mg), and 634 +/- 59 (12 mg) days], absence of ejaculate [416 +/- 88 (3 mg), 476 +/- 83 (6 mg), and 644 +/- 67 (12 mg) days], undetectable plasma concentrations of LH and testosterone [367 +/- 64 (3 mg), 419 +/- 72 (6 mg), and 607 +/- 69 (12 mg) days], the delay until complete recovery of LH and testosterone secretion [394 +/- 65 (3 mg), 484 +/- 72 (6 mg) and 668 +/- 47 (12 mg) days], and the delay until testes had regrown to normal volume [408 +/- 77 (3 mg), 514 +/- 74 (6 mg), 676 +/- 59 (12 mg) days]. The time taken to restore full ejaculates was also longest for the 12-mg dose: 716 +/- 67 (12 mg) days vs 440 +/- 66 (3 mg) and 538 +/- 83 (6 mg) days after implantation. There was no correlation between delay to recovery of normal ejaculate quality and body mass. We conclude that the dose-response relationship with deslorelin implants is not expressed with respect to the degree of suppression of reproduction, but on the maximum duration of suppression and thus to delay until recovery.

Liver phagocytosis was studied by means of electron microscopy in late stages of lethal canine endotoxin shock. A massive phagocytic activity was found within hepatic macrophages. The phagocytosed material found within Kupffer cells consisted of red cells, red-cell fragments, platelets, a fibrin-like material and damaged polymorphonuclear leukocytes. Red cells and red-cell fragments seemed to constitute the major part of the material involved.

Muscle responses to exercise are complex and include acute responses to exercise-induced injury, as well as longer term adaptive training responses. Using Alaskan sled dogs as an experimental model, changes in muscle gene expression were analyzed to test the hypotheses that important regulatory elements of the muscle's adaptation to exercise could be identified based on the temporal pattern of gene expression. Dogs were randomly assigned to undertake a 160-km run ( n = 9), or to remain at rest ( n = 4). Biceps femoris muscle was obtained from the unexercised dogs and two dogs at each of 2, 6, and 12 h after the exercise, and from three dogs 24 h after exercise. RNA was extracted and microarray analysis used to define gene transcriptional changes. The changes in gene expression after exercise occurred in a temporal pattern. Overall, 569, 469, 316, and 223 transcripts were differentially expressed at 2, 6, 12, and 24 h postexercise, respectively, compared with unexercised dogs (based on P ≤ 0.01 and an absolute fold change of ≥1.5). Increases in a number of known transcriptional regulators, including peroxisome proliferator-activated receptor-α, cAMP-responsive element modulator, and CCAAT enhancer binding protein-δ, and potential signaling molecules, including brain-derived neurotrophic factor, dermokine, and suprabasin, were observed 2 h after exercise. Biological functional analysis suggested changes in expression of genes with known functional relationships, including genes involved in muscle remodeling and growth, intermediary metabolism, and immune regulation. Sustained endurance exercise by Alaskan sled dogs induces coordinated changes in gene expression with a clear temporal pattern. RNA expression profiling has the potential to identify novel regulatory mechanisms and responses to exercise stimuli.

To investigate the kinetics in glucose metabolism, diabetic dogs were infused with double labelled glucose either when they were connected to an artificial beta cell after overnight insulin withdrawal (study I) or when they were disconnected from insulin supply after excellent metabolic control (study II). Fourteen hours after the last insulin injection, the animals had three-fold elevated rates of appearance Ra and of disappearance Rd of glucose in relation to non-diabetic controls; the metabolic clearance rate was reduced, glucose carbon recirculation was slightly elevated, and the % lactate from glucose was not altered. Glucosuria contributed approximately 30% to the elevated glucose turnover. In study I, Ra was normalized within 45 min after insulin supply but Rd increased transiently before returning to normal. In study II, plasma insulin was zero 30 min after termination of insulin supply. Ra increased immediately; Rd decreased slightly but increased thereafter. Lactate was elevated under all conditions. Its production from glucose increased slightly after initiation of insulin action. Glucose carbon recirculation was reduced to subnormal values when the animals were euglycemic but hyperinsulinemic.--It is concluded that even short intervals of relative lack of insulin action followed by restoration of glucose homeostasis, may induce wasting of substrates.

Cytochrome P-450 (CYP) epoxygenases and their arachidonic acid (AA) metabolites, the epoxyeicosatrienoic acids (EETs), have been shown to produce marked reductions in infarct size (IS) in canine myocardium either given before an ischemic insult or at reperfusion similar to that produced in ischemic preconditioning (IPC) and postconditioning (POC) protocols. However, no studies have addressed the possibility that EETs serve a beneficial role in IPC or POC. We tested the hypothesis that EETs may play a role in these two phenomena by preconditioning dog hearts with one 5-min period of total coronary occlusion followed by 10 min of reperfusion before 60 min of occlusion and 3 h of reperfusion or by postconditioning with three 30-s periods of reperfusion interspersed with three 30-s periods of occlusion. To test for a role of EETs in IPC and POC, the selective EET antagonists 14,15-epoxyeicosa-5(Z)-enoic acid (14,15-EEZE) or its derivative, 14,15-epoxyeicosa-5(Z)-enoic acid 2-[2-(3-hydroxy-propoxy)-ethoxy]-ethyl ester (14,15-EEZE-PEG), were administered 10 min before IPC, 5 min after IPC, or 5 min before POC. In a separate series, the selective EET synthesis inhibitor N-methylsulfonyl-6-(propargyloxyphenyl)hexanamide (MS-PPOH) was administered 10 min before IPC. Infarct size was determined by tetrazolium staining and coronary collateral blood flow at 30 min of occlusion and reperfusion flow at 3 h by radioactive microspheres. Both IPC and POC produced nearly equivalent reductions in IS expressed as a percentage of the area at risk (AAR) [Control 21.2 ± 1.2%, IPC 8.3 ± 2.2%, POC 10.1 ± 1.8% ( P < 0.001)]. 14,15-EEZE, 14,15-EEZE-PEG, and MS-PPOH markedly attenuated the cardioprotective effects of IPC and POC (14,15-EEZE and 14,15-EEZE-PEG) at doses that had no effect on IS/AAR when given alone. These results suggest a unique role for endogenous EETs in both IPC and POC.

The aim of the present study was to investigate how loading of cardiopulmonary baroreceptors induced by water immersion (WI) modifies baroreflex control of renal sympathetic nerve activity (RSNA) in conscious dogs. Nine dogs were chronically instrumented for measuring carotid sinus nerve activity (CSNA), RSNA, carotid arterial (Pca), and central venous (Pcv) pressures. The stimulus-response relationships of Pca-CSNA and Pca-RSNA were determined simultaneously in the same dog by changing Pca using rapid intravenous infusions vasoactive drugs during pre-WI and WI. WI increased central venous pressure significantly ( P < 0.05) by 10.4 mmHg. WI shifted the Pca-RSNA curve acutely leftward compared with the pre-WI period, which was associated with significant ( P < 0.05) decreases in the saturation pressure by 39.0 mmHg and operating range by 43.1 mmHg. WI relocated the operating pressure to near the saturation pressure, where the gain was low. The Pca-CSNA curve obtained during WI was identical to that obtained during pre-WI period. These results suggest that the shift in baroreflex control of RSNA could be attributed to the inhibitory influence of the cardiopulmonary mechanoreceptor loading and not by the resetting of carotid baroreceptors per se.

Background Natural orifice transluminal endoscopic surgery (NOTES) represents a potentially less-invasive alternative to conventional or laparoscopic surgery. Objective Our purpose was to develop a canine oophorectomy model for prospective evaluation of intraoperative complications, surgical stress, and postoperative pain and recovery with NOTES. Design Feasibility study. Setting Academic preclinical research. Patients Ten healthy female dogs. Interventions NOTES procedures were performed through gastric access with an electrocautery snare to resect and retrieve the ovaries. The gastrotomy was closed with prototype T-fasteners. Main Outcome Measurements Operative time; complications; postoperative pain scores, and nociceptive threshold; surgical stress markers (interleukin-6 [IL-6], C-reactive protein); systemic stress parameters (cortisol, glucose); necropsy evaluation at 10 to 14 days. Results The mean operative time was 154 minutes (SD ± 58 minutes) and no animals died as a result of complications from the procedure. The primary difficulty was incomplete ovarian excision and conversion to an open procedure in 1 dog. Serum glucose concentrations increased after surgery and remained elevated for at least 36 hours. The serum cortisol concentration was transiently increased from baseline at 2 hours after surgery. The serum IL-6 concentration peaked at 2 hours after surgery and returned to the baseline value by 18 hours. The serum C-reactive protein concentration increased significantly from baseline, peaked at 12 hours after surgery, and then slowly declined toward baseline but remained elevated at 72 hours after surgery. Nociceptive threshold measurements indicated increased sensitivity to pain for 2 to 24 hours after surgery. At necropsy, surgical sites were healing uneventfully with no significant damage to surrounding organs, no significant growth on bacterial cultures, and no evidence of peritonitis. Limitations Small number of animals, single center. Conclusions The NOTES approach to oophorectomy in dogs appears to be a reasonable alternative to traditional surgery. Attention must be paid to ensure complete excision of the ovaries.

We investigated whether rimonabant, a type 1 cannabinoid receptor antagonist, reduces visceral adipose tissue (VAT) and subcutaneous adipose tissue (SAT) in dogs maintained on a hypercaloric high-fat diet (HHFD). To determine whether energy expenditure contributed to body weight changes, we also calculated resting metabolic rate. Twenty male dogs received either rimonabant (1.25 mg·kg−1·day−1, orally; n = 11) or placebo ( n = 9) for 16 wk, concomitant with a HHFD. VAT, SAT, and nonfat tissue were measured by magnetic resonance imaging. Resting metabolic rate was assessed by indirect calorimetry. By week 16 of treatment, rimonabant dogs lost 2.5% of their body weight ( P = 0.029), whereas in placebo dogs body weight increased by 6.2% ( P < 0.001). Rimonabant reduced food intake ( P = 0.027), concomitant with a reduction of SAT by 19.5% ( P < 0.001). In contrast with the VAT increase with placebo ( P < 0.01), VAT did not change with rimonabant. Nonfat tissue remained unchanged in both groups. Body weight loss was not associated with either resting metabolic rate ( r2 = 0.24; P = 0.154) or food intake ( r2 = 0.24; P = 0.166). In conclusion, rimonabant reduced body weight together with a reduction in abdominal fat, mainly because of SAT loss. Body weight changes were not associated with either resting metabolic rate or food intake. The findings provide evidence of a peripheral effect of rimonabant to reduce adiposity and body weight, possibly through a direct effect on adipose tissue.

Background Enhancing plasma viscosity during fluid resuscitation results in vasodilation and improved microvascular perfusion in rodents subjected to hemorrhagic shock. We hypothesized that resuscitation with hyperviscous lactated Ringer's solution (hyperLRS) would result in improved tissue oxygenation and acid-base values in hemorrhaged dogs. Methods Twelve dogs were anesthetized and splenectomized. Vascular catheterization was performed, and tissue oxygen probes were placed in the jejunal serosa and skeletal muscle to assess macro- and microhemodynamic parameters. Baseline (BL) and posthemorrhage data were obtained. After 1 hour of hemorrhagic shock (mean arterial pressure [MAP] 30-40 mm Hg), treatment groups (n = 6) were administered bolus LRS or hyperLRS, and then received sufficient LRS to achieve and maintain an MAP between 60 mm Hg and 70 mm Hg. Data were obtained at 10, 30, 60, 120, and 180 minutes after fluid resuscitation. Results There were no significant differences between LRS or hyperLRS groups at BL or posthemorrhage. Blood and plasma viscosity were significantly increased by the administration of hyperLRS at all time points postresuscitation compared with LRS. Significantly more fluid was required to maintain MAP, and vascular hindrance was consistently lower in dogs administered hyperLRS versus LRS, suggesting viscosity-induced vasodilation. Central and mesenteric venous oxygen saturations were significantly decreased, whereas lactate and oxygen extraction ratios were significantly increased after hyperLRS administration compared with LRS. The tissue oxygen tension was similar in dogs administered hyperLRS or LRS. Conclusions A hyperviscous balanced electrolyte solution did not improve hemodynamic parameters, tissue oxygen tension, or acid-base values despite evidence for viscosity-induced vasodilation.

Since electric activity could be registered from various organs in the body, this study investigates the feasibility of recording such activity from the testicle. The testicles of 12 male dogs were exposed under anaesthesia and 3 electrodes were sutured to the tunica albuginea. In addition, transcutaneous recordings of the electric waves were performed. The effect of traumatic insult induced by testicular irradiation and by induction of hypogonadism on electric activity was studied. Spermatic cord clamping and orchidectomy was done to test their effect on the electric waves of the testicles. Electric waves were registered from the electrodes applied both directly and transcutaneously to the testicle. Each wave consisted of a negative followed by a positive deflection, with a mean frequency of 10.2±1.8 cycle s -1 and amplitude of 56.6 ± 8.4 μV. The waves recorded from the 6 electrodes applied (3 directly and 3 transcutaneously) in the same animal had similar amplitude and frequency readings. No abnormal waves were registered. After testicular irradiation or induction of hypogonadism, a 'silent' or 'dysrhythmic' electroorchidogram was obtained. Spermatic cord clamping did not change the normal electroorchidographic pattern. No waves were recorded after orchidectomy (silent electroorchidogram). To conclude, an electroorchidogram was configurated for the normal testicle. It showed changes after testicular insult. It is assumed that the electroorchidogram might reveal changes in the pathologic conditions of the testicle and this requires further study.

(-)-N-{2-[(R)-3-(6,7-Dimethoxy-1,2,3,4-tetrahydroisoquinoline-2-carbonyl)piperidino]ethyl}-4-fluorobenzamide (YM758), a novel "funny" If current channel (If channel) inhibitor, is developed as a treatment for stable angina and atrial fibrillation. In this study, the pharmacokinetic/pharmacodynamic (PK/PD) relationship after intravenous administration of YM758 to tachycardia-induced dogs was investigated and described based on the simplified compartment model. The PK of YM758 in dogs did not differ between the nontreated and tachycardia-induced groups. A drug-induced reduction in heart rate (HR) was clearly observed, and the half-life of the duration of the effect (approximately 4.0 h) was longer than that of the plasma concentration of the unchanged drug. The fitting and simulation procedure from the PK/PD relationship between the time profiles for YM758 plasma concentration and HR reduction had an ECe(50) value (YM758 concentration in the effective compartment resulting in a 50% decrease of the maximum effect) of 6.0 ng/ml, which did not agree with the results of the in vitro experiment using right atria isolated from guinea pigs (EC(30), 70.4 ng/ml). In addition, in the in vitro experiments, YM758 metabolites had a weak inhibitory effect, if any, on the spontaneous beat rate of the right atria from guinea pigs. These data, along with the previous finding that YM758 and its metabolites are eliminated rapidly from rat hearts, indicate that the duration of the pharmacological effect of YM758 (compared with the rapid elimination of the plasma drug concentration) may be the result of strong binding and/or slower dissociation of YM758 in the If channel. Such PK/PD analyses allow the pharmacological profiles of many drugs, especially cardiovascular drugs, to be more readily understood and better predicted during the clinical stages.

The elimination of felodipine in liver microsomes from dog and man were characterized by Km and Vmax. The results were compared with previous data reported for rat. In all species studied, felodipine was primarily metabolized to its corresponding pyridine analogue. The elimination rate order was rat greater than dog greater than man. The same species difference was observed in vivo for the oral plasma clearance which was; rat 26 1/hr/kg, dog 7.5 1/hr/kg and man 4.3 1/hr/kg. The intrinsic hepatic clearance of felodipine was predicted in vitro from Vmax over Km. The in vitro values were not significantly different from those observed in vivo. Felodipine is a high-clearance drug and the in vivo extraction ratios were about the same in all species: rat 0.80, dog 0.83 and man 0.84. The extraction ratios predicted from the in vitro studies, rat 0.91, dog 0.70 and man 0.80, agreed well with those observed in vivo.

The influences of different calcium-entry blockers, sialidase and caffeine on the biphasic contraction induced by prostaglandin (PG) F2 alpha in the feline basilar artery (BA) were studied in calcium-free medium. After incubation in calcium-free solution, PGF2 alpha induced a contraction of the BA amounting to 87% of the contraction in calcium-containing solution. The response was biphasic in 41 out of 42 vessel segments. PGF2 alpha-induced contractions were markedly attenuated in TRIS-buffered solutions as compared to contractions in Krebs solution. PGF2 alpha failed to induce a biphasic contraction (8 out of 9 preparations) in calcium-free HEPES-buffered solution. Calcium entry blockade with 1 mM manganese or 10(-5) M diltiazem abolished the second and major phase of the PGF2 alpha-induced contraction in calcium-free Krebs solution. The second contraction phase was also eliminated in four out of five preparations pretreated with sialidase (1 unit/ml for 30 min.), but was unaffected by a brief exposure to 20 mM caffeine in calcium-free medium. The present findings strongly support previous suggestions that a major part of the PGF2 alpha-induced contraction in calcium-free medium is mediated via the release of calcium bound to the exterior aspect of the cell membrane.

It is generally assumed that cerebrospinal fluid (CSF) is secreted in the brain ventricles, and so that after an acute blockage of the aqueduct of Sylvius an increase in the ventricular CSF pressure and dilatation of isolated ventricles may be expected. We have tested this hypothesis in cats after blocking the acqueduct, we measured the CSF pressure in both isolated ventricles and the cisterna magna and performed radiographic monitoring of the cross-sectional area of the lateral ventricle. The complete acqueductal blockage was achieved by implanting a plastic cannula into the aqueduct of Sylvius through a small tunnel in the vermis of the cerebellum in the chloralose anesthetized cats. After the reconstitution of the occipital bone, the CSF pressure was measured in the isolated ventricles via a plastic cannula implanted in the aqueduct of Sylvius and in the cisterna magna via a stainless steel cannula. During the following two hours the CSF pressures in the isolated ventricles and cisterna magna were identical to those in control conditions. We also monitored the ventricular cross-sectional are by means of radiography for two hours after the acqueductal blockage and failed to observe any significant changes. When mock CSF was infused into isolated ventricles to imitate the CSF secretion, the gradient of pressure between the ventricle and cisterna magna developed and disappeared as soon as the infusion was terminated. However, when mock CSF was infused into the cisterna magna at various rates, the resulting increased subarachnoid CSF pressure was accurately transmitted across the brain parenchyma into the CSF of isolated ventricles. The absence of an increase in the CSF pressure and ventricular dilatation during the two hours of aqueductal blockage suggest that a new reconsideration of the CSF physiology as well as of the development of acute hydrocephalus is required.

Mongrel dogs were given either whole blood (50 ml) or platelets prepared from 50 ml blood intravenously on three occasions before transplantation of a kidney from a different donor to that of the blood or platelets. All dogs were given azathioprine and prednisolone after transplantation. 60 × 108 platelets were obtained from 50 ml blood and the leucocyte contamination was less than one leucocyte per 105 platelets. The mean survival of kidney allografts in non-transfused dogs was 11.6 days (10 dogs), in dogs pretreated with whole blood 26.6 days (5 dogs), and in dogs pretreated with platelets 29.2 days (5 dogs). Sensitisation occurred in 3 of 5 dogs given whole blood and in 2 of 5 dogs given platelets. Thus pretreatment with a relatively pure preparation of platelets will produce prolongation of survival of third party renal allografts in mongrel dogs given azathioprine and prednisolone, comparable to that produced by whole blood. However, sensitisation was produced by the platelet preparation, presumably due to the minimal leucocyte contamination, which might also be responsible for the suppressive effect of this platelet pretreatment protocol.

This study was conducted to examine the regeneration process of hepatic nerves following surgical hepatic denervation in dogs. A denervation model was surgically created by removing all visible nerves around the hepatoduodenal ligament along with the peri-hepatic tissues. The hepatic nerves were examined on the hepatic specimens taken at 1 week, 1 month and 3 months post-denervation by means of immunohistochemical staining, and also electron microscopy. At 1 week post-denervation, the extrinsic hepatic nerves were observed not to have regenerated. However, at 1 month post-denervation, GAP-43-positive nerves were identified and regenerating axons were shown to be present on electron microscopic observation. The immunoreactivity for anti-GAP-43 antibody was not shown any longer at 3 months post-denervation, and the regenerated nerve axons were shown to be similar to those at pre-denervation on ultrastructural study. Results of the present study suggested that regeneration of the extrinsic hepatic nerves began to appear about 1 month after the hepatic denervation, and was completed by 3 months post-denervation.

The “push-pull” effect denotes the reduced tolerance to +Gz (hypergravity) when +Gz stress is preceded by exposure to hypogravity, i.e., fractional, zero, or negative Gz. The purpose of this study was to test the hypothesis that an exaggerated, myogenically mediated rise in leg vascular conductance contributes to the push-pull effect, using heart level arterial blood pressure as a measure of G tolerance. The approach was to impose control (30 s of 30° head-up tilt) and push-pull (30 s of 30° head-up tilt immediately preceded by 10 s of −15° head-down tilt) gravitational stress after administration of hexamethonium (5 mg/kg) to inhibit autonomic ganglionic neurotransmission in seven dogs. Cardiac output or thigh level arterial pressure (myogenic stimulus) was maintained constant by computer-controlled ventricular pacing. The animals were sedated with acepromazine and lightly restrained in lateral recumbency on a tilt table. Following the onset of head-up tilt, the magnitude of the fall in heart level arterial pressure from baseline was −11.6 ± 2.9 and −17.1 ± 2.2 mmHg for the control and push-pull trials, respectively ( P < 0.05), when cardiac output was maintained constant. Over 40% of the exaggerated fall in heart level arterial pressure was attributable to an exaggerated rise in hindlimb vascular conductance ( P < 0.05). Maintaining thigh level arterial pressure constant abolished the exaggerated rise in hindlimb blood flow. Thus a push-pull effect largely attributable to a myogenically induced rise in leg vascular conductance occurs when autonomic function is inhibited.

The rapid increase of incretins glucose-dependent insulinotropic peptide (GIP) and glucagon like peptide-1 (GLP-1), within 5-15 min, after food ingestion, suggests that a neural mechanism might be involved in the regulation of their secretion. The aim of this study is to determine whether intracerebroventricular (i.c.v) administration of neuropeptide Y (NPY), a widely distributed neurotransmmiter, can mediate this neural regulation of GIP secretion after food consumption. Six healthy mongrel dogs were utilized for this study. A prototype epicranial apparatus was placed surgically, allowing easy and exact localization of the third ventricle for infusions or sampling. Simultaneous blood sampling was obtained from cannulation of a hind limb vein. Plasma insulin, and GIP concentrations were measured after i.c.v infusion of 5, 10 and 25 microg of NPY dissolved in 0.5 ml of artificial cerebrospinal fluid (a CSF). The secretion of GIP and insulin were increased after the injection of NPY in a different pattern. Our data indicate that NPY might be involved in a possible neural control mechanism of GIP secretion after food consumption.