Role of ACE and ACE-2 in abrogated cardioprotective effect of ischemic preconditioning in ovariectomized rat heart

Abstract Ischemic heart disease is the leading cause of death in postmenopausal women. The activity of heart ACE increases whereas the activity of ACE-2 decreases after menopause. The present study was designed to investigate the role of ACE and ACE-2 in the abrogated cardioprotective effect of IPC in OVX rat heart. The heart was isolated from OVX rat and mounted on Langendorff's apparatus for giving intermittent cycles of IPC. The infarct size was estimated using TTC stain, and coronary effluent was analyzed for LDH, CK-MB, and nitrite release. IPC induced cardioprotection was significantly attenuated in the ovariectomized rat heart as compared to the normal rat heart. However, this attenuated cardioprotection was significantly restored by perfusion of DIZE, an ACE-2 activator, and captopril, an ACE inhibitor, alone or in combination noted in terms of decrease in myocardial infarct size, the release of LDH and CK-MB, and also increase in the release of NO as compared to untreated OVX rat heart. Thus, it is suggested that DIZE and captopril, alone or in combination restore the attenuated cardioprotective effect of IPC in OVX rat heart which is due to an increase in ACE-2 activity and decrease in ACE activity after treatment.

Development and validation of analytical method by HPLC-DAD for determination of vasodilator active in pharmaceutical ophthalmic forms

The substance 4-Aminobenzamidine dihydrochloride (4-AD) is one of the degradation products of diminazene aceturate and has demonstrated antiglaucomatous potential. Glaucoma is the second leading cause of blindness worldwide; thus, new therapeutic alternatives must be studied, for example, the molecule 4-AD vehiculated into polymeric inserts for prolonged release. The present work aims to develop and validate an analytical method to quantify 4-AD in pharmaceutical ophthalmic forms. A HPLC was used with UV-Vis detector, at 290 ƞm and ACE® C18 column (125 × 4.6 mm, 5 µm), in which the mobile phase consists of phosphate buffer (pH 7.4) and triethylamine (30 mmol/L), under an isocratic flow of 1.0 mL/min. The retention time of 3.2 minutes was observed. The method was developed and validated in accordance with ANVISA recommendations and ICH guides. The linearity range was established between the concentrations 5 and 25 µg/mL (correlation coefficient r = 0.993). The accuracy, repeatability, and intermediate precision tests obtained a relative standard deviation less than or equal to 5%. In addition, the method was considered selective, exact. and robust, with pH being its critical factor. Therefore, the HPLC analysis method is robust and can be used to quantify 4-AD in pharmaceutical forms for ocular application.(AU)

Protective effect of diminazene on kidney injury of limb ischemiareperfusion model mice and its mechanism / 吉林大学学报(医学版)

Objective: To detect the levels of angioteinsin II (Ang II) and angioteinsin (1-7) [Ang (1-7)] and the expression levels of angiotensin II type-1 receptor (AT1R) and Mas receptor (MasR) proteins in kidney tissue of the limb ischemia-reperfusion (LIR) mice pre-treated with the angiotensin coverting enzyme 2 (ACE2) activator diminazene (DIZE), and to explore the protective effect of DIZE on the kidney injury of the LIR mice. Methods: Eighteen male ICR mice aged 8 weeks were divided into control group, LIR group and LIR+DIZE group. The mice in model group and LIR+DIZE group were subjected to 2 h of ischemia and 4 h of reperfusion to establish the LIR models. The mice in LIR + DIZE group were pre-treated with 10 mg · kg-1 · d-1 DIZE for 14 d by subcutaneous injection before LIR. The histological technique was used to observe the morphology of kidney tissue of the mice and the pathological injury was evaluated. Chemical colorimetry was performed to determine the levels of serum urea and serum creatinine (Scr) of the mice. Enzyme linked immunosorbent assay (ELISA) was used to determine the Ang II and Ang (1-7) levels in kidney tissue of the mice. Western blotting method was used to measure the expression levels of AT1R and MasR proteins in kidney tissue of the mice. Results: Compared with control group, the pathological changes such as inflammatory cell infiltration and epithelial cell degeneration were found in kidney tissue of the mice in LIR group, and the kindey injury score was obviously increased (P<0. 05); compared with LIR group, the kidney injury performance in the kidey tissue of the mice in LIR + DIZE group was alleviated and the kidney injury score was decreased significantly (P<0. 05). Compared with control group, the levels of serum urea and Scr of the mice in LIR group were significantly increased (P<0. 05); compared with LIR group, the levels of serum urea and Scr of the mice in LIR + DIZE group were significantly decreased (P<0. 05). Compared with control group, the Ang II, Ang (1-7) levels and the ratio of Ang II/Ang (1-7) of the mice in LIR group were significantly increased (P<0. 05); compared with LIR group, the Ang II level of the mice in LIR+DIZE group was markedly decreased (P<0. 05), the Ang (1-7) level was significantly increased (P<0. 05)), and the ratio of Ang II/Ang (1-7) was decreased (P<0.05). Compared with control group, the expression level of AT1R protein in kidney tissue of the mice in LIR group was significantly decreased (P<0.05), the expression of MasR protein was significantly increased (P<0. 05), and the ATlR/MasR ratio was decreased (P<0.05). Compared with LIR group, the AT1R and MasR protein expression levels in kidney tissue of the mice in LIR + DIZE group were significantly increased (P < 0.05), and the ATlR/MasR ratio was also increased (P < 0.05). Conclusion: The imbalance of Ang II/Ang (1-7) and AT1R/Mas expressions in kidney tissue of the mice may be involved in kidney injury after LIR of the mice. ACE2 activitor DIZE may play a protective role in the kidney by improving the imbalance of Ang II/Ang (1-7) and ATlR/MasR expressions.

Efficacy of secnidazole-diminazene aceturate combination therapy in the late treatment of Trypanosoma brucei brucei infection in dogs

This study evaluated the efficacy of combination therapy of secnidazole-diminazene aceturate (SEC-DA) in late treatment of dogs experimentally infected with relapsing strain of Trypanosoma brucei brucei. Fifteen dogs were randomly assigned to 5 groups (A - E) of 3 per group. Group A (uninfected untreated), B (infected untreated), C (infected and treated with DA (3.5 mg/kg) IM stat), D (infected and treated with secnidazole (SEC) (100 mg/kg) orally for 5 days and DA (3.5 mg/kg) IM stat), E (infected and treated with SEC (200 mg/kg) orally for 5 days and DA (3.5 mg/kg) IM stat). Dogs were infected intraperitoneally with 5 x 105 trypanosomes and treatment started 14 days post-infection. Data on parasitaemia, hematology and rectal temperature were recorded. Parasitaemia cleared within 3 days in all the SEC-DA treated dogs and there was no relapse parasitaemia. Parasitaemia did not clear in DA monotherapy dogs. All the SEC-DA treated dogs showed significantly (P < 0.05) higher leucocyte counts, red blood cell count, packed cell volume, hemoglobin concentration and lower rectal temperature than DA monotherapy. It was, therefore, concluded that SEC-DA combination is therapeutically more efficacious than DA monotherapy in the late treatment of T.b. brucei infection in dogs.

Protective effect of DIZE on heart function of rats with diabetic cardio-myopathy / 中国病理生理杂志

AIM:To observed the protective effect of diminazene aceturate(DIZE),an angiotensin-converting enzyme 2(ACE2)activator,on rats with diabetic cardiomyopathy(DCM).METHODS:Male Wistar rats(n=30)were randomly divided into normal control group ,DCM group and DIZE treatment group(DIZE group).The rats in DCM group and DIZE group were intraperitoneally injected with streptozotocin(65 mg/kg )to establish diabetic model.After 12 weeks,the diabetic rats were infused with DIZE at 15 mg· kg-1 · d-1 or the same volume of saline for 4 weeks using os-motic minipump.The cardiac function was measured at the end of the 16th week.The methods of Mason staining and HE staining were used to observe the morphological changes of the myocardial tissue.Western blot ,ELISA and immunohisto-chemistry were used to observe the changes of ACE2,angiotensin(Ang)Ⅱ,Ang-(1-7),interleukin(IL)-1,IL-6 and connective tissue growth factor(CTGF).RESULTS:DIZE significantly improved the expression of ACE 2 in diabetic rats(P<0.05).Compared with DCM group,the levels of IL-1 and IL-6 in DIZE group were significantly decreased ,and the cardiac function in DIZE group was significantly improved(P<0.05).CONCLUSION:ACE2 endogenous agonist DIZE significantly increases the ACE 2 level and reduces the level of inflammation ,thus protecting the heart function of DCM rats.

Aspectos patológicos da intoxicação por aceturato de diminazeno em camelídeos sul-americanos / Pathological aspects of diminazene aceturate toxicosis in american camelids

Aceturato de diminazeno é um fármaco quimioterápico sintético comumente usado na medicina veterinária para o tratamento de doenças causadas por parasitos hematozoários. Entretanto, seu uso pode levar a efeitos colaterais, como alterações neurológicas graves e morte. A criação de camelídeos é uma atividade recente no Brasil, fazendo-se necessário conhecer mais sobre as doenças que acometem essas espécies. De dez camelídeos (seis lhamas e quatro alpacas) da propriedade, seis tiveram sinais clínicos e, destes, apenas uma lhama com manifestações leves recuperou-se. Os sinais clínicos incluíam apatia, andar cambaleante, fraqueza, sialorreia, cabeça baixa e pendida lateralmente, dificuldade em levantar e dispneia, observados a partir de 18 horas após o uso do medicamento. À necropsia e ao exame histopatológico foram observadas alterações de encefalopatia hemorrágica bilateral e simétrica, mais graves em tronco encefálico e tálamo. Este trabalho descreve as principais lesões observadas em um surto de intoxicação por diminazeno em alpacas (Lama pacos) e lhamas (Lama glama) e alerta criadores e veterinários sobre o risco de intoxicação por aceturato de diminazeno em camelídeos sul americanos.(AU)

Diminazene aceturate is a synthetic chemotherapeutic drug commonly used in veterinary medicine for the treatment of diseases caused by hematozoan parasites. However, side effects as severe neurological disorders and death can occur. The raising of american camelids is a recent activity in Brazil, requiring knowledge about diseases that affect these species, in order to avoid misguided conducts. In a herd of ten camelids (six llamas and four alpacas) six showed clinical signs and five died; only a llama with mild signs recovered. The clinical signs included apathy, difficulty to stand up, staggering gait, weakness, down head and drooping the head laterally, dyspnea and drooling of saliva, observed from 18 hours after use of the drug. At necropsy and histopathological examination was found bilateral and symmetrical hemorrhagic encephalopathy, more severe in brainstem and thalamus. This paper describes the main lesions observed in an outbreak of diminazene aceturate poisoning in alpacas (Lama pacos) and llamas (Lama glama) and alert breeders and veterinarians about the risk of poisoning by this drug in american camelids.(AU)

Preventive and therapeutic effect of diminazene in the rat model of pulmonary arterial hypertension / 中国比较医学杂志

Objective To explore the preventive and therapeutic effects of diminazene (DIZE) on pulmonary arterial hypertension (PAH) in rats.Methods Left pulmonary lobectomy combined with injection of monocrotaline was used to establish a rat model of pulmonary arterial hypertension.One hundred adult male Wistar rats were randomly divided into blank control group (group A,n=20),DIZE control group (group B,n=20),PAH model group (group C,n=20),PAH model plus DIZE group (group D,n=20) and PAH model plus DIZE and C-16 group (group E,n=20).Angiotensin-converting enzyme 2 (ACE2),IL-6 and IL-8 levels were determined by fluorescence resonance energy transfer (FRET) and enzyme linked immunosorbent assay (ELISA),and the mean pulmonary arterial pressure (mPAP) and right ventricular hypertrophy index (RVHI) were measured.The wall thickness (WT) and intimal hyperplasia score were calculated,and the pulmonary vascular lesions were analyzed using elastic fiber staining.Results RVHI,ACE2 enzyme activity and WT in the group A were significantly different from those of the groups C,D and E (P<0.05).Those of the group D and E were significantly different (P<0.05).The five groups showed significant differences in the overall analysis of each index (P<0.05).Conclusions Diminazene can increase the ACE2 enzyme activity,and decrease the mPAP,RVHI and WT,while reducing the pulmonary arterial medial hypertrophy,and inhibit intimal hyperplasia of pulmonary arterioles.The results of this study provide an experimental basis for the use of diminazene in the treatment of human pulmonary hypertension.

Protective effect of DIZE, an ACE2 activator, on rats with streptozoto-cin-induced diabetic nephropathy / 中国病理生理杂志

AIM:To observed the protective effect of diminazene aceturate ( DIZE) , an angiotensin-converting enzyme 2 (ACE2) activator, on diabetic nephropathy (DN) rats.METHODS:Male Wistar rats (n=30) were randomly divided into normal control (NC) group, DN group and DIZE group (each group consisted of 10 rats).The rats in DN group and DIZE group were induced by intraperitoneal injection of streptozotocin at dose of 65 mg/kg.After 12 weeks, the rats in DIZE group and DN group received subcutaneous injection of DIZE (15 mg· kg -1 · d-1 ) or vehicle for 4 weeks. The samples of blood and urine were collected at week 16, and ratio of kidney weight to body weight (KW/BW), plasma glucose (GLU), 24 h urinary protein (24UP) and serum creatinine (SCr) were measured.The renal pathological changes in each group were observed by periodic acid-Schiff ( PAS) staining and immunohistochemistry .The levels of AngⅡ and Ang-(1-7) in the plasma, and TGF-β1 and VCAM-1 in the renal tissues were measured by ELISA .The mRNA and protein levels of collagen I and FN were determined by quantification real-time PCR and immunohistochemistry .The effects of DIZE on the expression of ACE2 in DN rats were determined by Western blot .RESULTS:DIZE remarkably increased the expression of ACE2 and Ang-(1-7) in DN rats.Compared with NC group , the GLU, KW/BW, 24UP, SCr, and the ex-pression of collagen I , FN, TGF-β1 and VCAM-1 in DN group and DIZE group were increased .However , after treatment of the DN rats with DIZE, these indicators were decreased except the KW/BW.The GLU showed no significant change . CONCLUSION:DIZE raised the activity of ACE2 and increased the expression of Ang-(1-7), thus alleviating fibrosis and inflammation in the kidney and having therapeutic potential for diabetic nephropathy .

Blood Profil of Dogs Experimentally Infected with Single Trypanosoma brucei, Ancylostoma caninum and Conjunct Trypanosoma brucei and Ancylostoma caninum and Treatment with Diminazene Aceturate and Mebendazole.

The present study was designed to ascertain the level of haematological alterations in single Trypanosoma brucei (T. brucei), Ancylostoma caninum (A. caninum) and conjunct infections of both parasites in dogs and effect of treatment with diminazene aceturate and mebendazole on haematology. Sixteen dogs grouped into 4 of 4 members each were used in the study. Group 1 (GPI) was uninfected (control), GPII was infected with A. caninum, GPIII was infected with T. brucei and GPIV was infected with conjunct infections of T. brucei / A. caninum. Post acclimatization, GPII and GPIV were infected with A. caninum, 2 weeks after GPIII and GPIV were infected with T. brucei. By week 6 post infection, GPII and GPIV were treated with 100 mg of mebendazole given twice daily for 3 days and a repeat given 2 weeks later. GPIII and GPIV were also treated with diminazene aceturate at 7 mg/kg once. Treatment was repeated at week 8 and 9 of the experiment. There was a significant (p < 0.05) decreases in pack cell volume (PCV), haemoglobin concentration (Hb), red blood cell count (RBC) in all the experimental groups (GPII, GPIII and GPIV). The decreases were more in the conjunct group (GPIV) compared to the others. A significant (p < 0.05) decrease in white blood cell (WBC) count was recorded in all the experimental groups (GPII, GPIII and GPIV). It was reflected in significant (p <0.05) decreases in lymphocytes, neutrophil, monocyte, basophil counts in T. brucei infected group. Conversely there were significant (p <0.05) increases in neutrophil, eosinophil, monocyte and basophil count but a decrease in lymphocyte count in A. caninum group. The haematological alterations were more in T. brucie group compared to the A. caninum group. Similarly the effect was more in the conjunct T. brucei /A. caninum group compared to the single T. brucei. Treatment with 7 mg/kg diminazene aceturate and 100 mg mebendazole given once daily for 3 days caused some improvement in haematology. These findings would enhance clinicians’ knowledge of the effect of single and mixed infections of T. brucei and A. caninum in dogs.

Symptoms and Response to Treatment with Diminazene Aceturate and Mebendazole in Dogs Infected with Single Trypanosoma congolense, Ancylostoma caninum and Combination of Trypanosoma congolense and Ancylostoma caninum.

The economic losses associated with diseases caused by Trypanosoma congolense and the devastating effect of Ancylostoma caninum (A. caninum) in dogs’ necessitated the present study. Sixteen dogs grouped into 4 of 4 members each were used in the study. GROUP I was uninfected dogs (control), GROUP II was infected with Trypanosoma congolense (T. congolense) infection, GROUP III was mixed infections of Trypanosoma congolense and Ancylostoma caninum (T. congolense /A. caninum) and GPIV was infected with Ancylostoma caninum. At first Ancylostoma caninum infection was done on GPIII and GPIV. Two weeks later T. congolense infections was done on GPII and superimposed on GPIII. Three weeks post trypanosome infection; GPII and GPIII were treated with diminazene aceturate. Mebendazole was used on GPIII and GPIV and treatment repeated 2 weeks later. The prepatent period of T. congolense infection was 14.00±1.40 days in single infection and 9.00±1.10 days in conjunct infection of T. congolense and A. caninum. Persistent parasitaemia resulted in repeated treatment with diminazene aceturate at 7 mg/kg and mebendazole at 100mg twice daily for 3 days. The predominant signs revealed include; lethargy, vomition, enlargement of popliteal lymphnodes, pyrexia, oedema of fore and hind limbs and ocular discharges, anaemia, and slight emaciation. The symptoms were more severe in GPIII compared to GPII and GPIV. The egg per gram of faeces (EPG) in (GPIV) was significantly higher than the conjunct infection (GPIII). Treatment only slightly improved clinical manifestations. In conclusion, conjunct infections of T. congolense / A. caninum would result to more severe disease condition than in single infection of either disease in dogs. The severity of symptoms of the diseases were more in conjunct T. congolense / A. caninum as evidenced by high mortality compared with the single infections. Therefore symptoms of the diseases could serve as a surrogate diagnostic tool in diagnosis and vigorous treatment of infected dogs.

Clinical Effects of Mixed Infection of Trypanosomes and Ancylostoma caninum in Dogs and Treatment with Diminazene and Mebendazole (Nigeria).

The socio-economic importance of trypanosomosis and ancylostomosis in both humans and animal necessitated the investigation of the clinical signs of single and conjunct infection of both parasites in dogs. Sixteen dogs grouped into 4 of 4 members each were used in the study. GROUP I was uninfected dogs (control), GROUP II was infected with Ancylostoma caninum GROUP III was infected with Trypanosoma brucei (T. brucei), GROUP IV was mixed infections of Trypanosoma brucei and Ancylostoma caninum (T. brucei/A. caninum). Post acclimatization, Ancylostoma caninum infection was done on GPII and GPIV. Two weeks later Trypanosoma brucei infections was done on GPIII and superimposed on GPIV. Three weeks post trypanosome infection; GPIII and GPIV were treated with 7 mg/kg diminazene aceturate (Veribin®, CEVA Sante Animale- La Ballasteiére 33501 Libourne Cedex, France) x intramuscularly x once. Mebendazole (Vermin®, Janssen-Cilag Ltd 50 - 100 Holmers Farm Way, High Wycombe, Bucks, HP12 4EG UK) at 100 mg x per os twice daily for 3 days was used only on GPII and GPIV and a repeat treatment given 2 weeks later. Prepatent period of T. brucei infection was 5.00±1.30 days in single infection and 3.00±1.40 days in conjunct infection of T. brucei and A. caninum. Persistent parasitaemia resulted in repeated treatment with diminazene aceturate at 7 mg/kg and mebendazole at 100 mg twice daily for 3 days. The predominant signs revealed include; fluctuation in weight, lethargy, vomition, enlargement of popliteal lymphnodes, pyrexia, oedema of lower jaw and ocular discharges, enlarged abdomen, anaemia, cornea opacity and slight emaciation. The clinical signs were most severe in GPIV compared to GPIII. The egg per gram of faeces (EPG) in GPII was significantly higher than the mixed infection (GPIV). Treatment only slightly improved clinical manifestations. In conclusion, most signs shown were consistent with trypanosomosis in dogs except abdominal enlargement which is a complication of A. caninum. Clinical signs therefore could serve as a diagnostic tool in the treatment of both conditions in dogs.

Association of exercise training and angiotensin-converting enzyme 2 activator improves baroreflex sensitivity of spontaneously hypertensive rats

The present study sought to determine cardiovascular effects of aerobic training associated with diminazene aceturate (DIZE), an activator of the angiotensin converting enzyme 2, in spontaneously hypertensive rats (SHRs). Male SHRs (280–350 g) were either subjected to exercise training or not (sedentary group). The trained group was subjected to 8 weeks of aerobic training on a treadmill (five times a week, lasting 60 min at an intensity of 50–60% of maximum aerobic speed). In the last 15 days of the experimental protocol, these groups were redistributed into four groups: i) sedentary SHRs with daily treatment of 1 mg/kg DIZE (S+D1); ii) trained SHRs with daily treatment of 1 mg/kg DIZE (T+D1); iii) sedentary SHRs with daily treatment of vehicle (S+V); and iv) trained SHRs with daily treatment of vehicle (T+V). After treatment, SHRs were anesthetized and subjected to artery and femoral vein cannulation prior to the implantation of ECG electrode. After 24 h, mean arterial pressure (MAP) and heart rate (HR) were recorded; the baroreflex sensitivity and the effect of double autonomic blockade (DAB) were evaluated in non-anesthetized SHRs. DIZE treatment improved baroreflex sensitivity in the T+D1 group as compared with the T+V and S+D1 groups. The intrinsic heart rate (IHR) and MAP were reduced in T+D1 group as compared with T+V and S+D1 groups. Hence, we conclude that the association of exercise training with DIZE treatment improved baroreflex function and cardiovascular regulation.

An Assessment of Immune Response to Canine Distemper Vaccination in Dogs Experimentally Infected with Ancylostoma and Trypanosome Parasites.

The immunological alteration in vaccinated dogs with single hookworm, Ancylostoma caninum (A. c) and conjunct infection with Trypanosoma congolense (T. c) and Trypanosoma brucei (T. b) was determined. Sixteen dogs grouped into 4 of 4 members each were used. Group 1 was the uninfected control, GPII was infected with A. c, GPIII was infected with A. c /T. c, and GPIV was infected with T. b/A. c. The dogs were first inoculated with canine distemper (CD) vaccine before infection with A. c 4 weeks post vaccination. Two weeks later, both GPIII and GPIV were superposed with trypanosome infection. Prepatent period of A. c was 14 to 16 days in single A. c group and 13 to 14 days in conjunct trypanosome/A. c. The prepatent period of conjunct T. c/A. c was 9.00±1.10 days and 3.00±1.40 days, in conjunct T.bb/A. c. The protective antibody against CDV was considered using haemagglutination inhibition test (HIT) titer >100 as a cut off for seroconversion. At one week post vaccinations, the antibody titer against canine distemper (CDV) and anti-rabies in all the vaccinated groups (GPI, GPII, GPIII, and GPIV) significantly increased (p<0.05) and peaked at 3 weeks post vaccination. Subsequently, there was gradual significant decrease (p<0.05) in all the infected groups (GPII, GPIII and GPIV). The decrease in the conjunct groups (GPIII and GPIV) was higher compared to the single infections (GPII). Treatment with diminazene aceturate and mebendazole in all the groups did not significantly (p<0.05) improve antibody response in the dogs. A secondary vaccination administered at 12 weeks post- primary vaccination significantly increased (p<0.05) the antibody titer with a peak 3 weeks post- secondary vaccination. In conclusion, both trypanosomes and A. c induced primary immune suppression in antibody response to vaccination which improved on secondary vaccination in the infected dogs.

Aspectos epidemiológicos, clínicos e anatomopatológicos da intoxicação por aceturato de diminazeno em cães / Epidemiological, clinical, and pathological aspects of diminazene aceturate toxicosis in dogs

Os aspectos epidemiológicos, clínicos e anatomopatológicos da intoxicação espontânea por aceturato de diminazeno foram estudados em 10 cães. Em todos os casos, os cães afetados demonstraram sinais de síndrome tálamo-cortical, principalmente alteração do nível de consciência, tetraparesia, rigidez extensora e crise convulsiva. Em alguns casos, os cães acometidos apresentaram sinais de síndrome cerebelar, como tremores musculares generalizados de alta frequência e baixa amplitude, e/ou de síndrome vestibular, como ataxia, inclinação de cabeça e quedas. Esses sinais ocorreram entre 24 e 48 horas após o uso do fármaco injetável por via intramuscular e se mantiveram até a morte ou eutanásia dos cães (entre 1 e 7 dias). Tais sinais clínicos refletiam encefalomalacia hemorrágica focal simétrica, que afetava a medula oblonga, a ponte, a medular do cerebelo, o tálamo, o mesencéfalo, os pedúnculos cerebelares e os núcleos da base. Esse artigo: 1) descreve e discute essa forma de intoxicação medicamentosa tão pouco citada na literatura internacional e desconhecida da maior parte dos clínicos e patologistas veterinários brasileiros, 2) estabelece critérios clínicos e anatomopatológicos para o seu diagnóstico e, principalmente, 3) atenta para os riscos da utilização desse princípio ativo na terapêutica canina.

The epidemiological, clinical, and pathological aspects of diminazene aceturate (DA) spontaneous toxicosis were evaluated in 10 dogs. All affected dogs developed signs of thalamic-cortical syndrome, characterized mainly by neurological changes in the conscience levels, tetraparesis, extensor stiffness, and seizures. In some cases there was also evidence of cerebellar syndrome, characterized by generalized muscle tremors (high-frequency and low-amplitude) and/or vestibular syndrome, characterized by or ataxia, head tilt, and falling. These clinical signs occurred between 24 and 48 hours following intramuscular administration of DA and persisted until spontaneous death or euthanasia occurred between 1 and 7 days after the onset of clinical signs. The mentioned clinical signs reflected lesions that consisted of focal symmetrical hemorrhagic encephalomalacia affecting medulla oblongata, pons, cerebellar medulla, thalamus, midbrain, cerebellar peduncles, and basal nuclei. This article (1) describes and discusses DA toxicosis in dogs, a poorly-described clinical entity that is unknown by most clinicians and pathologists in Brazil; (2) establishes the clinical and pathological criteria for the diagnosis of DA toxicosis in dogs; and (3) calls up the attention for the risks of using DA in dogs in clinical settings.

Effect of diminazene aceturate, levamisole and vitamin C combination therapy in rats experimentally infected with Trypanosoma brucei brucei

OBJECTIVE@#To investigate the effect of diminazene aceturate (DA) alone or in combination with either levamisole and/or Vitamin C in albino rats experimentally infected with Trypanosoma brucei brucei.@*METHODS@#Thirty adult male albino rats, randomly assigned into 6 groups (A-F) of 5 rats each were used. They were either infected with 1×10(6) trypanosomes intraperitoneally (groups A-E) or uninfected (group F). The different groups were treated respectively as follows: group A-with 3.5 mg/kg DA; group B-3.5 mg/kg DA and 7.5 mg/kg levamisole; group C-3.5 mg/kg DA and 100 mg/kg vitamin C; and group D-3.5 mg/kg DA and 7.5 mg/kg levamisole and 100 mg/kg vitamin C. Group E was left untreated. Parameters assessed include: rectal temperature, body weight changes, packed cell volume (PCV), Haemoglobin concentration (Hb), total leucocyte count (TLC) differential leucocyte count (DLC), parasitaemia, clinical signs and survivability.@*RESULTS@#Average pre-patent period of 5 days was recorded. Parasites in the blood were cleared in all treated groups (A-D) within 48 hours post treatment (PT). Untreated rats in group E died between 25 and 32 days post infection (PI). Relapse was not recorded in all the treated groups (A-D). The initial reduction in PCV, Hb, TLC and increases in rectal temperature following infection were reversed by the treatments. The rats that received drug combinations (groups B, C and D) showed faster and higher recovery rates than the uninfected control and group A.@*CONCLUSIONS@#Levamisole and/or Vitamin C combination with DA were more effective in the treatment of rats infected with Trypanosoma brucei brucei.

Cerebellar encephalopathy from diminazene aceturate (beneril) toxicity in a dog / 대한수의학회지

A 2-year old castrated male Alaskan malamute was referred with primary complaints of marked anemia, hemeglobinuria and depression. Laboratory tests revealed canine babesiois with severe anemia. The dog was treated by blood transfusion and beneril (diminazene aceturate, 3.5 mg/kg IM). Two days after Beneril injection, the dog suddenly showed ataxia progressing to paresis. MRI revealed irregularly diffused lesions in the cerebellum. The case was tentatively diagnosed as cerebellar encephalopathy caused by diminazene toxicity. The dog successfully recovered following steroid therapy.

Cerebellar encephalopathy from diminazene aceturate (beneril) toxicity in a dog / 대한수의학회지

A 2-year old castrated male Alaskan malamute was referred with primary complaints of marked anemia, hemeglobinuria and depression. Laboratory tests revealed canine babesiois with severe anemia. The dog was treated by blood transfusion and beneril (diminazene aceturate, 3.5 mg/kg IM). Two days after Beneril injection, the dog suddenly showed ataxia progressing to paresis. MRI revealed irregularly diffused lesions in the cerebellum. The case was tentatively diagnosed as cerebellar encephalopathy caused by diminazene toxicity. The dog successfully recovered following steroid therapy.

Effects of Allium sativum Bulb Extract, Diminazene Aceturate and Their Combination on Parasitaemia, and Biochemical Indices in Rats Experimentally Infected with Trypanosoma Brucei Brucei.

Thirty five clinically healthy albino rats of both sexes weighing between 100 – 120grams were used to study the effects of Allium sativum bulb extract in combination with diminazene aceturate on parsitemia and biochemical indices in trypanosome brucei brucei infection. The rats were divided in to seven groups (A-G) of five rats each. All the infected rats developed Parasitemia five days post infection. Weakness, increase respiratory rate, rough hair coat Biochemical changes at interval and possible deaths were the major parameters which were carefully observed. All the treatment commenced at the onset of parasitemia by day five post infection. Sub therapeutic dose of Allium sativum at 20mg/kg/bw in combination with 1.7mg/kg/bw of diminazene aceturate (Group C),diminazene aceturate at single standard dose of 3.5mg/kg/bw (Group B) caused a significant reduction (P<0.05) in parasitemia. The liver function enzymes ALT AST level in rats infected and not treated showed significant increase liver function enzymes (Group A) while those treated with standard and sub therapeutic dose (Group BCD) respectively. Had their liver function enzymes towards normal, compare with control (Group G).Its trypanocidal activity was assessed through daily examination of blood parasite, sub therapeutic doses of Allium sativum bulb extracts and its combination appear to be more effective in reducing severity of trypanosome brucei brucei infection and provide alternative in reducing the toxicity of existing trypanocide.

Therapeutic effect of an anaplasmicid and anti-protozoa product againstthe blood parasites of cattle / Efecto terapéutico de un fármaco frente a los hemoparásitos del bovinoBabesia bovis, Babesia bigemina y Anaplasma marginale / Efeito terapêutico de um fármaco protoarizida e anaplasmicida frente aoshemoparasitos do bovino

In Colombia all cattle raising areas located in tropical areas of low and medium altitude are considered endemic for haemoparasites of cattle, but sometimes losses are minimized by the development of immunity and the achievementof a state of equilibrium in populations. Generally, in the field the use of effective drugs is required as a help in the control of clinical manifestations associated with the occurrence of these diseases. This study was aimed to validate the benefit of the use of an anaplasmicid and anti-protozoa product (Ganaplus ®) for the control of clinical events associated with blood parasites in cattle. The study consisted of a experimental phase based on artificialinoculation of cattle with Anaplasma marginale, Babesia bigemina and Babesia bovis; and a field phase that evaluated clinical events in animals raised in endemic areas. In both phases, blood sample collection and measurement of body temperature were conducted. Complete hemogram, hematocrit, and blood smears for parasite analysis wereperformed. Cattle of the experimental phase showed very low parasitemia, with slight changes in hematologicparameters, but the animals that demonstrated clinical acute states successfully responded to the application of the compound, restoring their hematologic parameters. The cattle of the field phase showed higher parasitemias, with clear reductions in the hematocrit and the presence of an acute clinical syndrome, which responded appropriately tothe product application. It was confirmed that the compound tested is a good medicine for the treatment of clinical disease associated with blood parasites.

En Colombia todas las zonas ganaderas localizadas en áreas de trópico bajo y trópico medio se consideran regionesenzoóticas para los hemoparásitos del ganado, pero en ocasiones las pérdidas se minimizan por el desarrollo deinmunidad y el alcance de un estado de equilibrio en las poblaciones. Generalmente, en el campo se requiere de fármacos eficaces que ayuden a controlar las manifestaciones clínicas que se asocian con la ocurrencia de estos hemoparasitismos. Esta investigación buscó validar la bondad del uso de un producto protozoaricida y anaplasmicida (Ganaplus®) para el control de episodios clínicos asociados con los hemoparásitos del ganado. El estudio constó de una fase experimental basada en inoculación artificial de bovinos, con Anaplasma marginale, Babesia bovis y Babesia bigemina; y una fase de campo que evaluó episodios clínicos en animales ubicados en zonas endémicas. En ambas fases se realizó recolección de muestras de sangre y medición de temperatura corporal. Se evaluaron cuadros hemáticos completos, hematocrito y los extendidos sanguíneos para análisis parasitológico. Los bovinos de la fase experimental presentaron parasitemias muy bajas, con leves cambios en los parámetros hematológicos, pero los animales que presentaron cuadros clínicos agudos respondieron de forma satisfactoria a la aplicación del compuesto restableciendo sus parámetros hematológicos. Los bovinos de la fase de campo presentaron parasitemias mayores, con reducciones evidentes en el hematocrito y la presencia de cuadros clínicos agudos, que respondieronadecuadamente a la aplicación del producto. Se confirmó que el compuesto evaluado es un buen medicamento parael tratamiento de cuadros clínicos asociados con enfermedad por hemoparásitos.

Na Colômbia todas as zonas da pecuária bovina em áreas do trópico baixo e trópico médio consideram-se regiõesendêmicas para os hemoparasitos do gado bovino, mas em ocasiões as perdas se minimizem pelo desenvolvimentode imunidade e o alcance de um estado de equilíbrio nas populações. Geralmente, no campo requerem-se de fármacoseficazes que controlem as manifestações clínicas que se associam com a ocorrência destes hemoparasitos. Esta pesquisa teve como objetivo validar a efetividade do uso de um produto protozoaricida e anaplasmicida (Ganaplus®)para o controle de episódios clínicos associados com os hemoparasitos bovinos. O estudo teve uma fase experimentalbaseada em inoculação artificial de bovinos com Anaplasma marginale, Babesia bigemina e Babesia bovis; euma fase de campo que avaliou episódios clínicos em animais localizados em zonas endêmicas. Nas duas fases realizou-se a colheita de amostras de sangue e a mensuração da temperatura corpórea. Avaliou-se a hematologia, hematocrito e esfregaços de sangue para análise parasitológica. Os bovinos da fase experimental apresentaramparasitemias baixas, com leves mudanças nos parâmetros hematológicos, mas os animais que apresentaramquadros clínicos agudos responderam satisfatoriamente à aplicação do produto, restabelecendo os seus parâmetroshematológicos. Os bovinos da fase de campo apresentaram parasitemias maiores, com reduções evidentes nohematocrito e a presença de quadros clínicos agudos, que responderam adequadamente à aplicação do produto.Confirmou-se que o composto avaliado é um bom medicamento para o tratamento de quadros clínicos associados com a doença por hemoparasitos.

In vitro and in vivo antileishmanial efficacy of a combination therapy of diminazene and artesunate against Leishmania donovani in BALB/c mice / Estudo in vitro e in vivo da eficácia anti leishmaniótica de terapêutica combinada de Diminazene e Artesunate contra Leishmania donovani em camundongos Balb/c

The in vitro and in vivo activity of diminazene (Dim), artesunate (Art) and combination of Dim and Art (Dim-Art) against Leishmania donovani was compared to reference drug; amphotericin B. IC50 of Dim-Art was found to be 2.28 ± 0.24 µg/mL while those of Dim and Art were 9.16 ± 0.3 µg/mL and 4.64 ± 0.48 µg/mL respectively. The IC50 for Amphot B was 0.16 ± 0.32 µg/mL against stationary-phase promastigotes. In vivo evaluation in the L. donovani BALB/c mice model indicated that treatments with the combined drug therapy at doses of 12.5 mg/kg for 28 consecutive days significantly (p < 0.001) reduced parasite burden in the spleen as compared to the single drug treatments given at the same dosages. Although parasite burden was slightly lower (p < 0.05) in the Amphot B group than in the Dim-Art treatment group, the present study demonstrates the positive advantage and the potential use of the combined therapy of Dim-Art over the constituent drugs, Dim or Art when used alone. Further evaluation is recommended to determine the most efficacious combination ratio of the two compounds.

A atividade in vitro e in vivo de Diminazene (Dim), Artezunate (Art) e a combinação Dim e Art (Dim-Art) contra Leishmania donovani foi comparada com a droga de referência Anfotericina B. IC50 da Dim-Art foi 2,28 ± 0,24 µg/mL enquanto aquelas de Dim e Art foram 9,16 ± 0,3 µg/mL e 4,64 ± 0,48 µg/mL respectivamente. O IC50 da Anfotericina B foi 0,16 ± 0,32 µg/mL contra a fase estacionária de promastigotas. A avaliação in vivo do modelo de L. donovani em camundongos Balb/c indicou que os tratamentos com a terapêutica de drogas combinadas em doses de 12,5 mg/kg por 28 dias consecutivos significantemente (p < 0,001) reduziu a carga parasitária no baço quando comparada a tratamentos com uma única droga dada nas mesmas dosagens. Embora a carga parasitária tenha sido levemente mais baixa (p < 0.05) no grupo Anfotericina B quando comparada com o grupo tratado Dim-Art, o estudo presente demonstra a vantagem positiva do uso potencial da terapêutica combinada Dim-Art sobre drogas como Dim ou Art quando usadas isoladamente. Posterior avaliação é recomendada para determinar a média de combinação mais eficaz dos dois compostos.