Exploration of the antiplatelet activity profile of betulinic acid on human platelets

Betulinic acid, a natural pentacyclic triterpene acid, presents a diverse mode of biological actions including antiretroviral, antibacterial, antimalarial, and anti-inflammatory activities. The potency of betulinic acid as an inhibitor of human platelet activation was evaluated, and its antiplatelet profile against in vitro platelet aggregation, induced by several platelet agonists (adenosine diphosphate, thrombin receptor activator peptide-14, and arachidonic acid), was explored. Flow cytometric analysis was performed to examine the effect of betulinic acid on P-selectin membrane expression and PAC-1 binding to activated platelets. Betulinic acid potently inhibits platelet aggregation and also reduced PAC-1 binding and the membrane expression of P-selectin. Principal component analysis was used to screen, on the chemical property space, for potential common pharmacophores of betulinic acid with approved antithrombotic drugs. A common pharmacophore was defined between the NMR-derived structure of betulinic acid and prostacyclin agonists (PGI2), and the importance of its carboxylate group in its antiplatelet activity was determined. The present results indicate that betulinic acid has potential use as an antithrombotic compound and suggest that the mechanism underlying the antiplatelet effects of betulinic acid is similar to that of the PGI2 receptor agonists, a hypothesis that deserves further investigation.     

Effects of animal position and number of repeated measurements on selected two-dimensional and M-mode echocardiographic variables in healthy dogs

OBJECTIVE: To evaluate the effects of positioning and number of repeated measurements on intra- and interobserver variability of echocardiographic measurements in dogs. DESIGN: Prospective study. ANIMALS: 4 healthy dogs. PROCEDURE: Each observer performed 24 examinations, separately assessing each dog 6 nonconsecutive times (3 times with the dog in lateral recumbency and 3 with the dog in a standing position). Variables evaluated included M-mode measurements of left ventricular end-diastolic and left ventricular end-systolic diameters, left ventricular free-wall thickness in diastole and systole, interventricular septal thickness in diastole and systole, left ventricular shortening fraction, and 2-dimensional measurements of the left atrial diameter-to-aortic diameter ratio. RESULTS: All coefficients of variation (range, 3.4% to 26.6%) were similar between operators and positions and were < 15% for 27 of 32 values. For both operators, repeatability of the measurements was better for left ventricular end-systolic diameter, left ventricular free-wall thickness in diastole, left ventricular free-wall thickness in systole, and the left atrial diameter-to-aortic diameter in the standing position, and similar for both positions for shortening fraction and left ventricular end-diastolic diameter. No effect of cardiac cycle was observed. CONCLUSIONS AND CLINICAL RELEVANCE: Within-day variability of conventional echocardiography performed with the dog in the standing position was at least as good as that obtained with the dog in lateral recumbency for most measured variables. Single measurements of each variable may be sufficient for trained observers examining dogs that do not have an arrhythmia. The standing position should be used, particularly for stressed or dyspneic dogs.     

Beating heart mitral valve replacement with a bovine pericardial bioprosthesis for treatment of mitral valve dysplasia in a Bull Terrier

OBJECTIVES: To describe an open, beating heart surgical technique and use of a bovine pericardial prosthetic valve for mitral valve replacement (MVR) in the dog. STUDY DESIGN: Clinical case report. ANIMALS: Male Bull Terrier (17-month-old, 26 kg) with mitral valve dysplasia and severe regurgitation. METHODS: A bovine pericardial bioprosthesis was used to replace the mitral valve using an open beating heart surgical technique and cardiopulmonary bypass. RESULTS: Successful MVR was achieved using a beating heart technique. Mitral regurgitation resolved and cardiac performances improved (left ventricular end-diastolic diameter decreased from 57.6 to 48.7 mm, and left atrium/aorta ratio returned to almost normal, from 1.62 to 1.19). Cardiopulmonary by-pass time and total surgical duration were decreased compared with standard cardioplegic techniques. Surgical recovery was uneventful and on echocardiography 6 months later valve function was excellent. CONCLUSION: Considering the technique advantages (no cardiac arrest, ischemic reperfusion injury, and hypothermia, or the need for aortic dissection and cannulation for administration of cardioplegic solution), short-term mortality and morbidity may be reduced compared with standard cardioplegic techniques. CLINICAL RELEVANCE: Based on experience in this dog, beating heart mitral valvular replacement is a seemingly safe and viable option for the dog and bovine pericardial prosthesis may provide better long-term survival than mechanical prostheses.     

Investigation of the antioxidant behavior of air- and freeze-dried aromatic plant materials in relation to their phenolic content and vegetative cycle

The total phenolic and flavonoid content of the aerial parts of five aromatic plants harvested at different periods was estimated, and their antioxidant capacity was evaluated. Major phenolic compounds present in their extracts were determined by RP-HPLC. The results demonstrated different amounts of total phenolic compounds and various degrees of antioxidant activity depending on the plant species, the time of harvest, and the drying method employed. Extracts from air-dried Mentha viridis L., Origanum majorana L., and Rosmarinus officinalis L. demonstrated the greatest efficacy during the flowering stage, in which the identified flavonoids were found in significantly higher amounts, whereas phenolic acids were found in their lowest concentration. Extracts from air-dried Laurus nobilis L. and Foeniculum vulgare Mill were less efficient in terms of antioxidant activity, with the highest values being observed during the early fruiting stage. This stage was characterized by the lowest flavonoid content and high phenolic acid content, except for L. nobilis L. extracts. Overall, the amount of identified phenolic acids did not vary considerably within the investigated year. The total phenolic concentration in all plant extracts decreased significantly when freeze-dried rather than air-dried samples were used. The HPLC analysis further supported the above for most of the phenolic compounds present in the extracts, except for hydroxybenzoic acids, which were better retained during the freeze-drying process.     

Effect of benazepril on survival and cardiac events in dogs with asymptomatic mitral valve disease: a retrospective study of 141 cases

BACKGROUND: Angiotensin-converting enzyme inhibitors (ACEIs) improve quality of life and extend the life span of dogs with naturally acquired ISACHC class II-III congestive heart failure (CHF). However, their effects on asymptomatic heart disease remain controversial. HYPOTHESIS: Benazepril (BNZ), an ACEI, could have beneficial effects at the asymptomatic stage of degenerative mitral valve disease (MVD). ANIMALS: Dogs with ISACHC class Ia MVD and moderate-to-severe mitral regurgitation (MR) assessed by the color Doppler mapping technique at entry (Day 0) were retrospectively included. METHODS: Dogs were assigned to the treated group (BNZ group) if they received BNZ (and no other cardiac medication) from Day 0 or to the untreated group (UT group) if they did not receive any cardioactive treatment until occurrence of CHF. RESULTS: A total of 141 dogs were included in the study, 66 in the BNZ group (dosage: 0.30 +/- 0.13 mg/kg) and 75 in the UT group. In the population (n = 93) including all breeds except Cavalier (CKC) and King Charles Spaniels (KC), median survival time to all causes of death in the BNZ group (n = 34, 3.3 years) was significantly longer than in the UT group (n = 59, 1.9 years) as was time to cardiac event (P < .05). Conversely, no effect of the BNZ treatment was observed in the CKC and KC population. CONCLUSIONS AND CLINICAL RELEVANCE: BNZ had beneficial effects in asymptomatic dogs other than CKC and KC affected by MVD with moderate-to-severe MR. Breed distribution should be taken into account for interpretation of clinical trials performed in dogs with cardiac disease.     

Chordae tendineae rupture in dogs with degenerative mitral valve disease: prevalence, survival, and prognostic factors (114 cases, 2001-2006)

Background:Degenerative mitral valve disease (MVD) is the most common heart disease in small breed dogs, and chordae tendineae rupture (CTR) is a potential complication of this disease. The survival time and prognostic factors predictive of survival in dogs with CTR remain unknown.
Hypothesis:The prevalence and prognosis of CTR in dogs with MVD increases and decreases, respectively, with heart failure class.
Animals:This study used 706 dogs with MVD.
Methods:The diagnosis of CTR was based on a flail mitral leaflet with the tip pointing into the left atrium during systole, which was confirmed in several 2-dimension imaging planes using the left and right parasternal 4-chamber views.
Results:CTR was diagnosed in 114 of the 706 dogs with MVD (16.1%) and most of these (106/114, 93%) had severe mitral valve regurgitation as assessed by color Doppler mode. CTR prevalence increased with International Small Animal Cardiac Health Council (ISACHC) clinical class (i.e., 1.9, 20.8, 35.5, and 69.6% for ISACHC classes Ia, Ib, II, and III, respectively [ P < .05]). Long-term follow-up was available for 57 treated dogs (angiotensin-converting enzyme inhibitors and diuretics) and 58% of these (33/57) survived > 1 year after initial CTR diagnosis (median survival time, 425 days). Clinical class, the presence of ascites or acute dyspnea at the time of diagnosis, heart rate, plasma urea concentration, and left atrial size were predictors of survival.
Conclusions and Clinical Relevance: CTR is associated with a higher overall survival time than previously supposed. Its prognosis mostly depends on a combination of clinical and biochemical factors.