Nephrin and Podocin mRNA Detection in Urine Sediment of Dogs with Chronic Kidney Disease: Preliminary Observations.

Introduction: Dogs with chronic kidney disease (CKD) may have alterations in the glomerular filtration barrier, including podocyte loss. Detection of podocyte mRNA in urine could be useful for assessing podocyturia in dogs with kidney disease. The objective of this study was to evaluate the presence of nephrin mRNA (NPHS1) and podocin mRNA (NPHS2) in urine sediments of dogs with naturally occurring CKD and healthy dogs. Material and Methods: Twenty-four dogs, 14 with CKD and 10 as healthy controls, underwent clinical evaluation. The dogs with CKD were divided into two groups, according to the International Renal Interest Society criteria: stage 1 or 2 CKD (n = 5) and stage 3 or 4 CKD (n = 9). Urine was collected by catheterisation or free catch and RNA isolation from the urine sediments was optimised using glycogen as a co-precipitant. Detection of NPHS1 and NPHS2 in the sediment samples was performed using quantitative real-time PCR. Results: Both types of mRNA were detected in samples from all groups, but the percentages of detection were higher in the group of dogs with stage 1 or 2 CKD and lower in the group of dogs with stage 3 or 4 disease. Conclusion: Physiological podocyturia was observed in healthy dogs, and the results suggest differential podocyturia in dogs with CKD, according to the stage of the disease, i.e. an increase in podocyturia in dogs at stage 1 or 2 and a reduction in podocyturia in dogs at stage 3 or 4.

Imbalance in the ratio between mineralocorticoid and glucocorticoid receptors and neurodegeneration in the dentate gyrus of aged dogs.

Background and Aim: Cortisol binds to mineralocorticoid receptor (MR) and glucocorticoid receptor (GR) found in the hippocampus. The balanced expression of these receptors is essential to neuronal survival as MR and GR activations have antiapoptotic and proapoptotic effects, respectively. Given the aging changes in dogs' dentate gyrus (DG) and the possible involvement of cortisol receptors in this process, this study aimed to evaluate the expression of MR and GR and neuronal degeneration in this hippocampal region of aged dogs. Materials and Methods: This study included cadaveric histologic hippocampus sections from six dogs aged 10 years and older (AG group) and 12 young/adult dogs aged up to 8 years (YAd group). Nissl staining and immunohistochemistry were performed to identify cells and investigate MR and GR expression, respectively. Furthermore, fluorescent labeling (fluoro-Jade B) was used to detect degenerating neurons. Results: The AG group's polymorphic layer of the DG had a lower cell count (16%) and more degenerating neurons than the YAd group. In addition to these cellular changes, the AG group had lower MR immunoreactivity and MR-to-GR ratio. Furthermore, the lowest MR expression was associated with neuronal degeneration in the polymorphic layer of the DG of dogs. Conclusion: An imbalance in the MR-to-GR ratio was observed in the polymorphic layer of the DG of aged dogs, along with lower MR expression and a greater number of degenerating neurons. These findings have clinical implications for understanding the decline in hippocampal memory formation associated with cognitive changes in aged dogs.

Evaluation of electrocardiographic parameters, thoracic morphometry and vertebral heart size in clinically normal dogs

Background: The Vertebral Heart Size (VHS) method sets standards for the evaluation of dogs heart size from the comparison of the cardiac dimension with the length of the thoracic vertebrae through radiographic study. Electrocardiogram(ECG) is widely used in veterinary medicine for the evaluation of electrical conduction system of the heart; however, theincrease of duration and amplitude of the ECG waves can suggest the increase of cardiac chambers in dogs. The scientificliterature presents electrocardiographic and VHS values for dogs of different breeds and sizes; however, there is little information on the correlation of these parameters. Therefore, the aim of this study was to evaluate the amplitude and durationof ECG waves, thoracic morphometry and VHS values, in order to correlate these parameters in clinically normal dogs.Materials, Methods & Results: Twenty healthy dogs (11 females and 9 males), without breed distinction, medium sized(14.46 ± 2.92 kg) and aged between 1 and 8 years, were evaluated through physical examination, digital ECG (frontaland precordial leads) and thorax X-ray in right lateral (RL), left lateral (LL) and ventrodorsal (VD) projections. Thoracicmorphometry and VHS measurements were determined as previously described. Clinical and ECG parameters were compatible with the references in all dogs evaluated. Dogs presented thorax intermediate (75%) with the depth and width ratio(D/W ratio) > 1.0. Some individual VHS values were higher than the references and the mean VHS values in VD projectionwas higher than in RL and LL projections (n = 20; P < 0.05). Female and male dogs did not differ among the evaluatedparameters. A positive correlation was observed between thoracic morphometry and body weight (r ≥ +0.70; P < 0.001)and a negative correlation was found between the D/W ratio and VD VHS (r = -0.62; P < 0.05). No significant correlationswere observed between the age, ECG parameters, thoracic morphometry and...(AU)

Evaluation of coagulation parameters in dogs with overweight or obesity

Background: Hemostatic alterations have already been well characterized in humans with body fat excess, being includedin the list of obesity related comorbidities. Overweight and obesity are common in dogs; however, there is little informationabout the blood coagulation parameters in dogs with these conditions. The aim of this study was to compare hematologicaland coagulation parameters between lean dogs and overweight or obese dogs, including platelets count; prothrombin time(PT); activated partial thromboplastin time (aPTT), coagulation time (CT) and plasma fibrinogen concentration.Materials, Methods & Results: A total of 22 dogs (aged 1 to 10 years, neutered) were evaluated, 10 of them presentedideal body condition score (BCS 4-5) and formed the group 1 (control) and 12 were overweight or obese (BCS 7-9) andformed the group 2. The dogs were submitted to clinical evaluation and then to blood collection for the following laboratorytests: blood count (performed on automatic analyzer), quantification...

Evaluation of electrocardiographic parameters, thoracic morphometry and vertebral heart size in clinically normal dogs

Background: The Vertebral Heart Size (VHS) method sets standards for the evaluation of dog’s heart size from the comparison of the cardiac dimension with the length of the thoracic vertebrae through radiographic study. Electrocardiogram(ECG) is widely used in veterinary medicine for the evaluation of electrical conduction system of the heart; however, theincrease of duration and amplitude of the ECG waves can suggest the increase of cardiac chambers in dogs. The scientificliterature presents electrocardiographic and VHS values for dogs of different breeds and sizes; however, there is little information on the correlation of these parameters. Therefore, the aim of this study was to evaluate the amplitude and durationof ECG waves, thoracic morphometry and VHS values, in order to correlate these parameters in clinically normal dogs.Materials, Methods & Results: Twenty healthy dogs (11 females and 9 males), without breed distinction, medium sized(14.46 ± 2.92 kg) and aged between 1 and 8 years, were evaluated through physical examination, digital ECG (frontaland precordial leads) and thorax X-ray in right lateral (RL), left lateral (LL) and ventrodorsal (VD) projections. Thoracicmorphometry and VHS measurements were determined as previously described. Clinical and ECG parameters were compatible with the references in all dogs evaluated. Dogs presented thorax intermediate (75%) with the depth and width ratio(D/W ratio) > 1.0. Some individual VHS values were higher than the references and the mean VHS values in VD projectionwas higher than in RL and LL projections (n = 20; P < 0.05). Female and male dogs did not differ among the evaluatedparameters. A positive correlation was observed between thoracic morphometry and body weight (r ≥ +0.70; P < 0.001)and a negative correlation was found between the D/W ratio and VD VHS (r = -0.62; P < 0.05). No significant correlationswere observed between the age, ECG parameters, thoracic morphometry and...

Evaluation of coagulation parameters in dogs with overweight or obesity

Background: Hemostatic alterations have already been well characterized in humans with body fat excess, being includedin the list of obesity related comorbidities. Overweight and obesity are common in dogs; however, there is little informationabout the blood coagulation parameters in dogs with these conditions. The aim of this study was to compare hematologicaland coagulation parameters between lean dogs and overweight or obese dogs, including platelets count; prothrombin time(PT); activated partial thromboplastin time (aPTT), coagulation time (CT) and plasma fibrinogen concentration.Materials, Methods & Results: A total of 22 dogs (aged 1 to 10 years, neutered) were evaluated, 10 of them presentedideal body condition score (BCS 4-5) and formed the group 1 (control) and 12 were overweight or obese (BCS 7-9) andformed the group 2. The dogs were submitted to clinical evaluation and then to blood collection for the following laboratorytests: blood count (performed on automatic analyzer), quantification...(AU)

Efeito da tiletamina-zolazepam e morfina em ratos: período de latência, avaliação bioquímica e hematológica / Effect of tiletamine-zolazepam and morphine in rats: latency period, biochemistry and hematological evaluation

Os pesquisadores têm buscado alternativas para minimizar a dor e o estresse dos animais de laboratório. Os procedimentos que causam desconforto e dor devem ser realizados com o uso de anestesia e analgesia. A associação tiletamina-zolazepam (TZ) é utilizada para a anestesia de roedores, tendo como vantagens a fácil administração e rápida indução, e as desvantagens da longa recuperação e analgesia variável. Nesta situação, o uso da morfina poderia proporcionar uma analgesia adicional; porém, os efeitos da TZ e a morfina ainda não foram descritos em ratos. O objetivo deste estudo foi determinar o período de latência de um protocolo envolvendo a TZ (30 mg/Kg) e a morfina (2 mg/Kg)em ratos Wistar machos, além de avaliar parâmetros bioquímicos e hematológicos desta espécie, após a anestesia. A TZ e a morfina foram administradas, conjuntamente, por via intramuscular. O sangue foi obtido por punção cardíaca e para os testes laboratoriais foram utilizadas técnicas já padronizadas. Os resultados indicaram que o protocolo anestésico foi eficaz para a anestesia e analgesia nos ratos, devido ao curto período de latência (6,26 ±1,23 min, N = 50) e a ausência de respostas aos estímulos dolorosos. Exceto pelo maior valor de ureia, a TZ e a morfina, praticamente, não interferiram nos parâmetros bioquímicos; porém foram observadas alterações hematológicas importantes, como: baixos valores para a contagem de hemácias e a concentração de hemoglobina.

Researchers seek alternatives to minimize the pain and stress of laboratory animals. Procedures that promote pain should be performed under anesthesia and analgesia. The combination tiletamine-zolazepam (TZ) is used for rodent anesthesia, with advantages: easy administration and rapid induction and disadvantages: long recovery and variable analgesia. In this situation, morphine could provide an additional analgesia; however, the effects of TZ plus morphine have not been described in rats. The objective of this study was to determine the latency period of TZ plus morphine in rats Wistar male, and evaluate the biochemical and hematological parameters of this species, after anesthesia. TZ plus morphine were administered intramuscularly. Blood was obtained by cardiac puncture and laboratory tests were performed using standardized techniques. The results indicated that the anesthetic protocol was effective to anesthesia and analgesia in rats, due to the short latency period (6.26 ± 1.23 min, N = 50) and absence of responses to painful stimuli. Except for the higher urea, TZ plus morphine, practically, did not interfere in the biochemical parameters; but important hematological alterations were observed: low values for red blood cell count and hemoglobin concentration.

Efeito da tiletamina-zolazepam e morfina em ratos: período de latência, avaliação bioquímica e hematológica / Effect of tiletamine-zolazepam and morphine in rats: latency period, biochemistry and hematological evaluation

Os pesquisadores têm buscado alternativas para minimizar a dor e o estresse dos animais de laboratório. Os procedimentos que causam desconforto e dor devem ser realizados com o uso de anestesia e analgesia. A associação tiletamina-zolazepam (TZ) é utilizada para a anestesia de roedores, tendo como vantagens a fácil administração e rápida indução, e as desvantagens da longa recuperação e analgesia variável. Nesta situação, o uso da morfina poderia proporcionar uma analgesia adicional; porém, os efeitos da TZ e a morfina ainda não foram descritos em ratos. O objetivo deste estudo foi determinar o período de latência de um protocolo envolvendo a TZ (30 mg/Kg) e a morfina (2 mg/Kg)em ratos Wistar machos, além de avaliar parâmetros bioquímicos e hematológicos desta espécie, após a anestesia. A TZ e a morfina foram administradas, conjuntamente, por via intramuscular. O sangue foi obtido por punção cardíaca e para os testes laboratoriais foram utilizadas técnicas já padronizadas. Os resultados indicaram que o protocolo anestésico foi eficaz para a anestesia e analgesia nos ratos, devido ao curto período de latência (6,26 ±1,23 min, N = 50) e a ausência de respostas aos estímulos dolorosos. Exceto pelo maior valor de ureia, a TZ e a morfina, praticamente, não interferiram nos parâmetros bioquímicos; porém foram observadas alterações hematológicas importantes, como: baixos valores para a contagem de hemácias e a concentração de hemoglobina.(AU)

Researchers seek alternatives to minimize the pain and stress of laboratory animals. Procedures that promote pain should be performed under anesthesia and analgesia. The combination tiletamine-zolazepam (TZ) is used for rodent anesthesia, with advantages: easy administration and rapid induction and disadvantages: long recovery and variable analgesia. In this situation, morphine could provide an additional analgesia; however, the effects of TZ plus morphine have not been described in rats. The objective of this study was to determine the latency period of TZ plus morphine in rats Wistar male, and evaluate the biochemical and hematological parameters of this species, after anesthesia. TZ plus morphine were administered intramuscularly. Blood was obtained by cardiac puncture and laboratory tests were performed using standardized techniques. The results indicated that the anesthetic protocol was effective to anesthesia and analgesia in rats, due to the short latency period (6.26 ± 1.23 min, N = 50) and absence of responses to painful stimuli. Except for the higher urea, TZ plus morphine, practically, did not interfere in the biochemical parameters; but important hematological alterations were observed: low values for red blood cell count and hemoglobin concentration.(AU)