Preactivation of neutrophils and systemic oxidative stress in dogs with hyperleptinemia.

High occurrence of obesity currently constitutes the main nutritional disease of the canine species. There is evidence that leptin increases during obesity in dogs. Hyperleptinemia is associated with increased neutrophil oxidative metabolism in obese humans and contributes to oxidative stress. However, in obese dogs, the probable relationship between this condition and the activation of the oxidative metabolism of neutrophils has yet to be established. Thus, we investigated the hypothesis that neutrophil activation and systemic oxidative stress occur in dogs with hyperleptinemia. A control group of 24 healthy dogs with a body condition score (BCS) of 4-5, an overweight group of 25 dogs with a BCS of 6-7, and 27 obese dogs with a BCS of 8-9, were composed. Two subgroups were formed composed of dogs with and without hyperleptinemia, grouped according to the 95% confidence interval obtained for plasma leptin values of the control group. Changes in obesity markers (body condition score, adiponectin and plasma leptin) and plasma oxidative stress (lipid peroxidation, total antioxidant and oxidant capacities and oxidative stress index) were measured in all the dogs selected. Neutrophil oxidative metabolism was evaluated in flow cytometry by superoxide production with the probe hydroethidine and by hydrogen peroxide production with the probe 2',7'-dichlorofluorescein diacetate, with or without phorbol myristate acetate (PMA) stimulation. Apoptosis and neutrophil viability were quantified in a capillary flow cytometer using Annexin VPE, with or without camptothecin apoptosis inducing effect. Obese dogs presented higher systemic oxidative stress, hyperleptinemia and preactivated neutrophils with accelerated apoptosis. Dogs with hyperleptinemia and obese dogs presented higher neutrophil superoxide production under PMA stimulation and the presence of systemic oxidative stress compared with control. To our knowledge, this is probably the first evidence that preactivation of the oxidative metabolism of circulating neutrophils occurs in dogs with hyperleptinemia, a condition that can induce systemic oxidative stress in the canine species.

Correlations between peripheral parasite load and common clinical and laboratory alterations in dogs with visceral leishmaniasis.

Intensity of peripheral parasite infection has an important role in the transmission of Leishmania spp. from one host to another. As parasite load quantification is still an expensive procedure to be used routinely in epidemiological surveillance, the use of surrogate predictors may be an important asset in the identification of dogs with high transmitting ability. The present study examined whether common clinical and laboratory alterations can serve as predictors of peripheral parasitism in dogs naturally infected with Leishmania spp. Thirty-seven dogs were examined in order to establish correlations between parasite load (PL) in multiple peripheral tissues and common clinical and laboratory findings in canine visceral leishmaniasis (CVL). Quantitative polymerase chain reaction was employed to determine PL in conjunctival swabs, ear skin, peripheral blood and buffy coat. Additionally, a series of hematological, biochemical and oxidative stress markers were quantified. Correlations between net peripheral infection and severity of clinical alterations and variation in laboratory parameters were assessed through a new analytical approach, namely Compressed Parasite Load Data (CPLD), which uses dimension reduction techniques from multivariate statistics to summarize PL across tissues into a single variable. The analysis revealed that elevation in PL is positively correlated with severity of clinical sings commonly observed in CVL, such as skin lesions, ophthalmic alterations, onycogriphosis, popliteal lymphadenomegaly and low body mass. Furthermore, increase in PL was found to be followed by intensification of non-regenerative anemia, neutrophilia, eosinopenia, hepatic injury and oxidative imbalance. These results suggest that routinely used clinical and laboratory exams can be predictive of intensity of peripheral parasite infection, which has an important implication in the identification of dogs with high transmitting ability.

Estresse oxidativo em cães com doença periodontal: comparação dos biomarcadores plasmáticos e salivares / Oxidative stress in dogs with periodontal disease: comparison of plasmatic and salivary biomarkers

A doença periodontal (DP) é a enfermidade inflamatória mais comum da cavidade oral dos cães. A quantificação de biomarcadores do plasma e da saliva tem sido utilizada para avaliar o estresse oxidativo sistêmico (EOS) e local (EOL) da DP humana. Na DP canina, os mecanismos do estresse oxidativo não estão bem caracterizados e estabelecidos. O objetivo do presente estudo foi investigar a hipótese de que o EOS ocorre na DP canina e de que a saliva pode ser utilizada para avaliar o EOL. Analisou-se, também, a hipótese de que a ativação do metabolismo oxidativo dos neutrófilos contribui para EOS na DP dos cães. Para tal, foram selecionados 20 cães adultos portadores de DP, agrupados de acordo com o grau de lesão: gengivite (n=6), periodontites leve (n=8) e avançada (n=6). O grupo controle foi composto pelos mesmos 20 cães, 30 dias após o tratamento periodontal. Para avaliar o metabolismo oxidativo dos neutrófilos circulantes foi quantificada a produção de superóxido pelo teste de redução do tetrazólio nitroazul (NBT). As concentrações de oxidante total (TOC) e de espécies reativas ao ácido tiobartbitúrico (TBARS) no plasma foram quantificadas para avaliar o EOS. Para a avaliação do estresse oxidativo local, foi quantificado o TOC salivar e a concentração dos principais antioxidantes da saliva (albumina, ácido úrico e bilirrubina total). O EOS na DP foi confirmado pelo aumento da produção de superóxido dos neutrófilos circulantes, TOC e TBARS plasmático. Foi possível quantificar todos os biomarcadores na saliva de cães, porém nenhum foi capaz de expressar o EOL da DP canina. Esta é uma das primeiras evidências de que o EOS ocorre em cães com DP e que a ativação do metabolismo oxidativo dos neutrófilos pode contribuir para desequilíbrio entre antioxidantes e oxidantes. Este estudo ressalta a importância da higiene bucal dos cães para a prevenção da DP e de lesões degenerativas crônicas de diversos tecidos causadas pelo EOS.(AU)

Periodontal disease (PD) is the most common inflammatory disease of the oral cavity of dogs. Quantitation of plasma and salivary biomarkers have been used to assess the systemic oxidative stress (SOS) and local (LOS) of human PD. In canine PD, oxidative stress mechanisms are not well characterized and established. Our objective was to investigate the hypothesis that SOS occurs in dog PD and saliva can be used to evaluate the LOS. We also investigated the hypothesis that the activation of neutrophil oxidative metabolism contributes to SOS in dog SD. For this purpose, 20 adult dogs were selected PD patients, grouped according to the degree of injury: gingivitis (n=6), light periodontitis (n=8) and advanced periodontitis (n=6). The control group was composed of the same 20 dogs, 30 days after periodontal treatment. To assess oxidative metabolism of circulating neutrophils superoxide production was measured by test nitroblue tetrazolium reduction (NBT). The total oxidant concentrations (TOC) and reactive species to tiobartbitúrico acid (TBARS) in plasma were quantified to evaluate SOS. For the evaluation of local oxidative stress were quantified salivary TOC and concentration of the main antioxidant in saliva (albumin, uric acid, and total bilirubin). EOS in dogs with PD was confirmed by increased superoxide production of circulating neutrophils, TOC, and plasma TBARS. It was possible to quantify all the biomarkers in the saliva of dogs, but none was able to express the LOS canine PD. This is the first evidence that SOS occurs in dogs with PD and that activation of the oxidative metabolism of neutrophils may contribute to an imbalance between oxidants and antioxidants. This study highlights the importance of oral hygiene of dogs to prevent PD and chronic degenerative lesions of various tissues caused by SOS.(AU)