Equilíbrio acidobásico, parâmetros urinários e sanguíneos de gatos induzidos ao estresse e suplementados com composto antioxidante / Acid-base balance, urinary and blood parameters of cats stress-induced cats and supplemented with antioxidant compound

Este estudo teve como objetivo avaliar a suplementação do composto antioxidante comercial EconomasE (Alltech, Brasil ( AOX) sobre o equilíbrio acidobásico, os parâmetros urinários, o hemograma completo e a fragilidade osmótica de eritrócitos (FOE) de gatos estressados. Foram utilizados 24 gatos adultos (3,49±0,87kg), distribuídos em delineamento inteiramente ao acaso, com quatro níveis (0, 250, 500, 750mg de AOX/kg de alimento na matéria seca) e seis repetições, durante o período de 80 dias. Os gatos foram induzidos ao estresse por meio da presença de cães próximo ao recinto experimental (agente estressor; AE), do 61º dia até o final do experimento. A pressão parcial de dióxido de carbono (pCO2) e o bicarbonato (HCO3 -) aumentaram linearmente (P<0,05) conforme o aumento da ingestão de AOX. Os maiores valores de dióxido de carbono total (tCO2) (P<0,05) foram observados nos gatos alimentados com 500 e 750mg de AOX/kg de dieta. As concentrações de hemoglobina foram maiores nos animais alimentados com 250 e 500mg de AOX/kg de dieta. Os parâmetros urinários e da FOE foram semelhantes nos gatos em todas as dietas. Esses dados indicam que a suplementação com AOX apresenta efeitos benéficos no equilíbrio acidobásico e na concentração de hemoglobina de gatos induzidos ao estresse.(AU)

This study aimed to evaluate the supplementation of a commercial antioxidant EconomasE (Alltech, Brazil - AOX) on the acid-base balance, urinary parameters, red blood count (RBC) and erythrocyte osmotic fragility (EOF) of stressed cats. Twenty-four adult cats (3.49±0.87kg) were distributed in a completely randomized design with four levels (0, 250, 500, 750mg AOX/kg feed dry matter) and six replicates for a period of 80 days. Cats were induced to stress through the presence of dogs in the experimental environment (stressor agent; AE) from the 61th day to the end. Partial pressure of carbon dioxide (pCO2) and bicarbonate (HCO3) increased linearly (P<0.05) with increased intake of AOX. The highest values of total carbon dioxide (tCO2) (P<0.05) were observed in cats fed the 500 and 750mg AOX/kg diet. Hemoglobin concentration was higher (P<0.05) in animals fed the 250 and 500mg AOX/kg diet. The urinary parameters and EOF were similar among all diets. These data indicate that AOX supplementation has beneficial effects in acid-base balance and hemoglobin concentration of stress-induced cats.(AU)

Equilíbrio acidobásico, parâmetros urinários e sanguíneos de gatos induzidos ao estresse e suplementados com composto antioxidante / Acid-base balance, urinary and blood parameters of cats stress-induced cats and supplemented with antioxidant compound

Este estudo teve como objetivo avaliar a suplementação do composto antioxidante comercial EconomasE (Alltech, Brasil ( AOX) sobre o equilíbrio acidobásico, os parâmetros urinários, o hemograma completo e a fragilidade osmótica de eritrócitos (FOE) de gatos estressados. Foram utilizados 24 gatos adultos (3,49±0,87kg), distribuídos em delineamento inteiramente ao acaso, com quatro níveis (0, 250, 500, 750mg de AOX/kg de alimento na matéria seca) e seis repetições, durante o período de 80 dias. Os gatos foram induzidos ao estresse por meio da presença de cães próximo ao recinto experimental (agente estressor; AE), do 61º dia até o final do experimento. A pressão parcial de dióxido de carbono (pCO2) e o bicarbonato (HCO3 -) aumentaram linearmente (P<0,05) conforme o aumento da ingestão de AOX. Os maiores valores de dióxido de carbono total (tCO2) (P<0,05) foram observados nos gatos alimentados com 500 e 750mg de AOX/kg de dieta. As concentrações de hemoglobina foram maiores nos animais alimentados com 250 e 500mg de AOX/kg de dieta. Os parâmetros urinários e da FOE foram semelhantes nos gatos em todas as dietas. Esses dados indicam que a suplementação com AOX apresenta efeitos benéficos no equilíbrio acidobásico e na concentração de hemoglobina de gatos induzidos ao estresse.(AU)

This study aimed to evaluate the supplementation of a commercial antioxidant EconomasE (Alltech, Brazil - AOX) on the acid-base balance, urinary parameters, red blood count (RBC) and erythrocyte osmotic fragility (EOF) of stressed cats. Twenty-four adult cats (3.49±0.87kg) were distributed in a completely randomized design with four levels (0, 250, 500, 750mg AOX/kg feed dry matter) and six replicates for a period of 80 days. Cats were induced to stress through the presence of dogs in the experimental environment (stressor agent; AE) from the 61th day to the end. Partial pressure of carbon dioxide (pCO2) and bicarbonate (HCO3) increased linearly (P<0.05) with increased intake of AOX. The highest values of total carbon dioxide (tCO2) (P<0.05) were observed in cats fed the 500 and 750mg AOX/kg diet. Hemoglobin concentration was higher (P<0.05) in animals fed the 250 and 500mg AOX/kg diet. The urinary parameters and EOF were similar among all diets. These data indicate that AOX supplementation has beneficial effects in acid-base balance and hemoglobin concentration of stress-induced cats.(AU)

Morfometria intestinal em codornas de corte alimentadas com treonina digestível / Intestinal morphometry in meat quails fed with digestible threonine

The aim of the study was to evaluate intestinal morphology data by including different levels of threonine in the diet of meat quails in the growing phase. The experimental design was completely randomized in a factorial arrangement: four nutritional levels and three intestinal segments with ten repetitions. Samples were collected to obtain morphometry data (villi and crypt) of the three segments of the small intestine (duodenum, jejunum, and ileum). In the study of the intestinal villus height a highly significant interaction was obtained for all ages studied. A crypt depth was evidenced at 42 days with a significant interaction between levels of inclusion of threonine in the diet and small intestine. In the study of the relationship villus height and depth crypt quails at 21 days of age, the results were significant for the interaction treatment (T) and part of the intestine. At 21 and 42 days of age, supplementation of 1.04 and 0.78% respectively of threonine in the diet promotes better outcomes for intestinal morphology.(AU)

O objetivo deste estudo foi avaliar a morfometria intestinal por meio da inclusão de níveis de treonina digestível na dieta de codornas para corte em crescimento. O delineamento experimental adotado foi o inteiramente ao acaso, em esquema fatorial: quatro níveis nutricionais e três segmentos do intestino com dez repetições. Foram coletadas amostras dos três segmentos do intestino delgado. As variáveis estudadas foram área e altura das vilosidades intestinais, profundidade de cripta e relação vilo e cripta. No estudo da altura da vilosidade, foi obtida interação significativa para todas as idades estudadas. A profundidade da cripta em codornas de corte aos 21 dias de idade mostrou-se significativa para as variáveis inclusão de treonina na dieta e para a porção do intestino estudada. Na profundidade da cripta aos 42 dias, ficou evidenciada uma interação significativa entre níveis de inclusão de treonina na dieta e porção do intestino delgado. No estudo da relação altura de vilosidade e profundidade de cripta em codornas aos 21 dias de idade, foram encontrados resultados significativos para a interação tratamento e porção do intestino. Aos 21 e aos 42 dias de idade, a suplementação de 1,04 e 0,78%, respectivamente, de treonina digestível na dieta promove melhores resultados para morfometria intestinal.(AU)

Morfometria intestinal em codornas de corte alimentadas com treonina digestível / Intestinal morphometry in meat quails fed with digestible threonine

O objetivo deste estudo foi avaliar a morfometria intestinal por meio da inclusão de níveis de treonina digestível na dieta de codornas para corte em crescimento. O delineamento experimental adotado foi o inteiramente ao acaso, em esquema fatorial: quatro níveis nutricionais e três segmentos do intestino com dez repetições. Foram coletadas amostras dos três segmentos do intestino delgado. As variáveis estudadas foram área e altura das vilosidades intestinais, profundidade de cripta e relação vilo e cripta. No estudo da altura da vilosidade, foi obtida interação significativa para todas as idades estudadas. A profundidade da cripta em codornas de corte aos 21 dias de idade mostrou-se significativa para as variáveis inclusão de treonina na dieta e para a porção do intestino estudada. Na profundidade da cripta aos 42 dias, ficou evidenciada uma interação significativa entre níveis de inclusão de treonina na dieta e porção do intestino delgado. No estudo da relação altura de vilosidade e profundidade de cripta em codornas aos 21 dias de idade, foram encontrados resultados significativos para a interação tratamento e porção do intestino. Aos 21 e aos 42 dias de idade, a suplementação de 1,04 e 0,78%, respectivamente, de treonina digestível na dieta promove melhores resultados para morfometria intestinal.(AU)

The aim of the study was to evaluate intestinal morphology data by including different levels of threonine in the diet of meat quails in the growing phase. The experimental design was completely randomized in a factorial arrangement: four nutritional levels and three intestinal segments with ten repetitions. Samples were collected to obtain morphometry data (villi and crypt) of the three segments of the small intestine (duodenum, jejunum, and ileum). In the study of the intestinal villus height a highly significant interaction was obtained for all ages studied. A crypt depth was evidenced at 42 days with a significant interaction between levels of inclusion of threonine in the diet and small intestine. In the study of the relationship villus height and depth crypt quails at 21 days of age, the results were significant for the interaction treatment (T) and part of the intestine. At 21 and 42 days of age, supplementation of 1.04 and 0.78% respectively of threonine in the diet promotes better outcomes for intestinal morphology.(AU)

Inclusion of Yucca schidigera and zeolite in dog feed: influence on mineral excretion / Inclusão de Yucca schidigera e zeólita em alimentos para cães: influência sobre a excreção de minerais

This study aimed to evaluate the effects of adding Yucca schidigera and zeolite (Clinoptilolite) to feed (super premium and standard) for adult dogs on the apparent indigestibility coefficient of minerals. In the first assessment, 21 dogs were used, with average weight of 12.5kg±1.46 in seven treatments in two periods (super premium diet referred to as control and control with the addition of 125, 250, 375ppm Yucca schidigera and 0.5%, 0.75% and 1.0% zeolite). The second assessment was conducted with standard feed, using 21 experimental plots distributed in three treatments (standard diet referred to as control and control with the addition of 375ppm of Yucca schidigera and 1.0% zeolite). The additives did not affect the acceptability of feed and fecal characteristics in both assessments (P>0.05). With the exception of calcium, no minerals were affected by the inclusion of additives (P>0.05) in the experiment with super premium feed. In the assessment with standard feed, calcium as phosphorus and magnesium presented decreased excretion (P<0.05) with the inclusion of additives. It is possible to conclude that the inclusion of additives in the tested concentrations may interfere with the excretion of some minerals in the diet.(AU)

Este trabalho teve como objetivo avaliar os efeitos da inclusão de Yucca schidigera e zeólita (Clinoptilolita) em alimentos (super premium e standard) para cães adultos sobre o coeficiente de indigestibilidade aparente de minerais. No primeiro ensaio, foram utilizados 21 cães, com peso médio de 12,5kg±1,46, distribuídos em sete tratamentos em dois períodos (dieta super premium denominada controle e controle com adição de 125, 250, 375ppm de Yucca schidigera e 0,5%, 0,75% e 1,0% de zeólita). O segundo foi realizado com um alimento standard, utilizando-se 21 parcelas experimentais distribuídas em três tratamentos (dieta standard denominada controle e controle com adição de 375ppm de Yucca schidigera e 1,0% de zeólita). Os aditivos não afetaram a aceitabilidade do alimento e as características fecais nos dois ensaios (P>0,05). Com exceção do cálcio, nenhum mineral sofreu interferência da inclusão dos aditivos (P>0,05) no experimento com alimento super premium. No ensaio com alimento standard, tanto cálcio quanto fósforo e magnésio apresentaram redução na excreção (P<0,05) com a inclusão dos aditivos. Pôde-se concluir que a inclusão dos aditivos nas concentrações testadas pode interferir na excreção de alguns minerais da dieta.(AU)

Inclusion of Yucca schidigera and zeolite in dog feed: influence on mineral excretion / Inclusão de Yucca schidigera e zeólita em alimentos para cães: influência sobre a excreção de minerais

This study aimed to evaluate the effects of adding Yucca schidigera and zeolite (Clinoptilolite) to feed (super premium and standard) for adult dogs on the apparent indigestibility coefficient of minerals. In the first assessment, 21 dogs were used, with average weight of 12.5kg±1.46 in seven treatments in two periods (super premium diet referred to as control and control with the addition of 125, 250, 375ppm Yucca schidigera and 0.5%, 0.75% and 1.0% zeolite). The second assessment was conducted with standard feed, using 21 experimental plots distributed in three treatments (standard diet referred to as control and control with the addition of 375ppm of Yucca schidigera and 1.0% zeolite). The additives did not affect the acceptability of feed and fecal characteristics in both assessments (P>0.05). With the exception of calcium, no minerals were affected by the inclusion of additives (P>0.05) in the experiment with super premium feed. In the assessment with standard feed, calcium as phosphorus and magnesium presented decreased excretion (P<0.05) with the inclusion of additives. It is possible to conclude that the inclusion of additives in the tested concentrations may interfere with the excretion of some minerals in the diet.

Este trabalho teve como objetivo avaliar os efeitos da inclusão de Yucca schidigera e zeólita (Clinoptilolita) em alimentos (super premium e standard) para cães adultos sobre o coeficiente de indigestibilidade aparente de minerais. No primeiro ensaio, foram utilizados 21 cães, com peso médio de 12,5kg±1,46, distribuídos em sete tratamentos em dois períodos (dieta super premium denominada controle e controle com adição de 125, 250, 375ppm de Yucca schidigera e 0,5%, 0,75% e 1,0% de zeólita). O segundo foi realizado com um alimento standard, utilizando-se 21 parcelas experimentais distribuídas em três tratamentos (dieta standard denominada controle e controle com adição de 375ppm de Yucca schidigera e 1,0% de zeólita). Os aditivos não afetaram a aceitabilidade do alimento e as características fecais nos dois ensaios (P>0,05). Com exceção do cálcio, nenhum mineral sofreu interferência da inclusão dos aditivos (P>0,05) no experimento com alimento super premium. No ensaio com alimento standard, tanto cálcio quanto fósforo e magnésio apresentaram redução na excreção (P<0,05) com a inclusão dos aditivos. Pôde-se concluir que a inclusão dos aditivos nas concentrações testadas pode interferir na excreção de alguns minerais da dieta.

TLR4 and RAGE: similar routes leading to inflammation in type 2 diabetic patients.

AIM: The present study investigates the interaction of TLR4 and RAGE with their respective ligands as inducers of the inflammatory markers IL-6 and TNF-α. Also, the reactivity of peripheral blood mononuclear cells (PBMNC) from type 2 diabetic (T2D) patients and non-diabetic healthy controls (ND) were comparatively studied. METHODS: Concentrations of IL-6 and TNF-α were measured by sandwich Elisa, using kits supplied by Assay Designs (Ann Arbor, MI, USA). PBMNC from T2D and ND were incubated in the presence or absence of LPS, anti-TLR4 or anti-RAGE for 72 hours at 37°C under 5% CO(2). The final volume was adjusted to 300 µL in DMEM supplemented with 10% fetal bovine serum. After incubation, the cells were centrifuged, the supernatant collected and the cytokines measured. RESULTS: PBMNC from T2D were more sensitive to innate immune stimulation with LPS and monoclonal agonist anti-TLR4 than were cells from ND. The actions of LPS, anti-TLR4 and anti-RAGE potentiated the production of IL-6 and TNF-α in both groups. The simultaneous activation of monoclonal anti-RAGE and anti-TLR4 suggests that both antibodies used different receptors on the cell surface, but converged on the same PBMNC signaling metabolic pathways. This simultaneous activation induced a higher production of IL-6 and TNF-α in PBMNC from the T2D patients than from the ND subjects. CONCLUSION: Our results clearly show an exacerbation of innate immunity in PBMNC with T2D that was possibly hyperglycaemia-induced. These data, when analyzed together, suggest the importance of innate immunity in the pathogenesis of T2D.

Inhibition of ROS production in peripheral blood mononuclear cells from type 2 diabetic patients by autologous plasma depends on Akt/PKB signaling pathway.

OBJECTIVE: To compare the role of Akt/PKB signaling pathway in the modulation of reactive oxygen species (ROS) production by autologous plasma in peripheral blood mononuclear cells (PBMNC) from type 2 diabetic patients and healthy subjects. MATERIALS AND METHODS: This study was approved by Santa Casa Ethical Committee and has included patients diagnosed with diabetes type 2 (DM2) and control group (non-diabetic) (ND). PBMNC were purified utilizing Ficoll-hypaque gradient. ROS was quantified by luminol-dependent chemiluminescence. The Akt/PKB phosphorylation was measured using a CASE kit. Statistical analyses were made with t Student test and chi-square (chi(2)). p<0.05 was considered significant. RESULTS: 12, 13-Phorbol dibutyrate (PDB) stimulated the production of higher levels of ROS in PBMNC from type 2 diabetic patients than that from healthy subjects. Autologous plasma, however, inhibited induced or not ROS production in PBMNC in both groups. The inhibition of PBMNC-ROS derived by autologous plasma from healthy subjects was higher than that from type 2 diabetic patients. Plasma phosphorylated (activated) Akt/PKB. The percentage of phosphorylation induced by autologous plasma in PBMNC from patients and healthy control were 14% and 93%, respectively. Inhibition of ROS production in PBMNC from DM2 were similar for PBMNC+plasma; PBMNC+Akti; and PBMNC+plasma+Akti. However, in ND control, plasma showed a higher ROS inhibition than Akti or plasma plus Akti. CONCLUSIONS: Our results suggest that the low antioxidant capacity observed in autologous plasma from DM2 patients in conjunction with the decreased activation of PKB may cause an imbalance in the oxidizing/reducing responses, possible inducing an oxidative stress state, which could be associated with tissular damage.

Changes in cell shape and desmin intermediate filament distribution are associated with down-regulation of desmin expression in C2C12 myoblasts grown in the absence of extracellular Ca2+.

Desmin is the main intermediate filament (IF) protein of muscle cells. In skeletal muscle, desmin IFs form a scaffold that interconnects the entire contractile apparatus with the subsarcolemmal cytoskeleton and cytoplasmic organelles. The interaction between desmin and the sarcolemma is mediated by a number of membrane proteins, many of which are Ca2+-sensitive. In the present study, we analyzed the effects of the Ca2+ chelator EGTA (1.75 mM) on the expression and distribution of desmin in C2C12 myoblasts grown in culture. We used indirect immunofluorescence microscopy and reverse transcription polymerase chain reaction (RT-PCR) to analyze desmin distribution and expression in C2C12 cells grown in the presence or absence of EGTA. Control C2C12 myoblasts showed a well-spread morphology after a few hours in culture and became bipolar when grown for 24 h in the presence of EGTA. Control C2C12 cells showed a dense network of desmin from the perinuclear region to the cell periphery, whereas EGTA-treated cells showed desmin aggregates in the cytoplasm. RT-PCR analysis revealed a down-regulation of desmin expression in EGTA-treated C2C12 cells compared to untreated cells. The present results suggest that extracellular Ca2+ availability plays a role in the regulation of desmin expression and in the spatial distribution of desmin IFs in myoblasts, and is involved in the generation and maintenance of myoblast cell shape.

Changes in cell shape and desmin intermediate filament distribution are associated with down-regulation of desmin expression in C2C12 myoblasts grown in the absence of extracellular Ca2+

Desmin is the main intermediate filament (IF) protein of muscle cells. In skeletal muscle, desmin IFs form a scaffold that interconnects the entire contractile apparatus with the subsarcolemmal cytoskeleton and cytoplasmic organelles. The interaction between desmin and the sarcolemma is mediated by a number of membrane proteins, many of which are Ca2+-sensitive. In the present study, we analyzed the effects of the Ca2+ chelator EGTA (1.75 mM) on the expression and distribution of desmin in C2C12 myoblasts grown in culture. We used indirect immunofluorescence microscopy and reverse transcription polymerase chain reaction (RT-PCR) to analyze desmin distribution and expression in C2C12 cells grown in the presence or absence of EGTA. Control C2C12 myoblasts showed a well-spread morphology after a few hours in culture and became bipolar when grown for 24 h in the presence of EGTA. Control C2C12 cells showed a dense network of desmin from the perinuclear region to the cell periphery, whereas EGTA-treated cells showed desmin aggregates in the cytoplasm. RT-PCR analysis revealed a down-regulation of desmin expression in EGTA-treated C2C12 cells compared to untreated cells. The present results suggest that extracellular Ca2+ availability plays a role in the regulation of desmin expression and in the spatial distribution of desmin IFs in myoblasts, and is involved in the generation and maintenance of myoblast cell shape.

Ribavirin, but not interferon alpha-2b, is associated with impaired osteoblast proliferation and differentiation in vitro.

Hepatitis C treatment with interferon alpha-2b (IFN-alpha) and ribavirin has been related to decreased bone mineral density. The aim of this study was to investigate the in vitro effects of different concentrations of ribavirin and IFN-alpha on osteoblast-like cells. Human osteoblast-like cells obtained by the outgrowth of cells from bone chips were exposed to ribavirin (0.1-10 microg/mL) or IFN-alpha (0.1-1000 UI/mL). At regular time-points, cultures were harvested for posterior analysis. Alkaline phosphatase (ALP) activity was determined on days 7 and 14, and cell growth was accessed by C3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) and cell count on days 1, 3, 5, and 7. Flow cytometry analysis was used for investigating cell death on days 1, 3, 5, and 7. IFN-alpha affected ALP expression only at the higher concentration (1000 UI/mL) after 7 days (P < 0.05). No effects were detected in cell growth. In ribavirin treated cultures, concentrations higher than 2.5 microg/mL were associated with a decrease in ALP activity within 7 and 14 days (P < 0.01 and P < 0.001, respectively). Furthermore, the reduction in cell growth was dose-dependent and was detected after the fifth day. This decrease can be explained by an increase in the number of dead cells and a decrease in cell proliferation. In conclusion, our experiments demonstrated that ribavirin reduced, in a time- and dose-dependent manner, the number of metabolically active cells through a decrease in proliferation and an increase in cell death, and induced an impairment in osteoblast differentiation. These negative effects of ribavirin on osteblast-like cells might contribute to the bone loss reported in vivo.