Oxidonitrergic and antioxidant effects of a low molecular weight peptide fraction from hardened bean (Phaseolus vulgaris) on endothelium

About 3000 tons of beans are not used in human food due to hardening. Several studies on bean-derived bioactive peptides have shown potential to treat some diseases, including those relying on oxidative dysfunctions. We assessed the effects of peptides extracted from hardened bean Phaseolus vulgaris (PV) on reactive oxygen species (ROS) and nitric oxide (NO) production, cytotoxic and cytoprotective effects in endothelial cells, and oxidonitrergic-dependent vasodilating effects. Extract was composed by peptide fraction <3 kDa (PV3) from hardened common bean residue. PV3 sequences were obtained and analyzed with bioinformatics. Human umbilical vein endothelial cells were treated with 10, 20, 30, and 250 µg/mL PV3. Oxidative stress was provoked by 3% H2O2. Cytotoxicity and cytoprotective effects were evaluated by MTT assay, whereas, ROS and NO were quantified using DHE and DAF-FM fluorescent probes by confocal microscopy. NO- and endothelium-dependent vasodilating effects of PV3 were assessed in isolated aortic rings. We found 35 peptides with an average mass of 1.14 kDa. There were no cell deaths with 10 and 20 μg/mL PV3. PV3 at 30 μg/mL increased cell viability, while cytotoxicity was observed only with 250 μg/mL PV3. PV3 at 10 μg/mL was able to protect cells from oxidative stress. PV3 also increased NO release without causing cell death. It also reduced relative ROS production induced by H2O2. PV3 vasodilating effects relied on endothelium-dependent NO release. PV3 obtained from low-commercial-value bean displays little cytotoxicity and exerts antioxidant effects, whereas it increases endothelial NO release.

Avaliação proteica do humor aquoso de equinos / Evaluation of equine aqueous humor protein

A avaliação proteica do humor aquoso (HA) pode ser utilizada como método diagnóstico nas uveítes. Entretanto, estudos sobre as proteínas nesse fluido, em equinos hígidos, são escassos e apresentam variações conforme a metodologia empregada. Dessa forma, objetivou-se realizar a análise proteica e citológica do HA nessa espécie, bem como verificar sua correlação com as proteínas plasmáticas. Foram avaliados 13 equinos adultos (26 olhos), sem raça definida, machos ou fêmeas. Mediante aqueocentese, foi coletado 0,5 mL de humor aquoso de cada olho. Cada amostra foi encaminhada para quantificação proteica pelo método de Bradford modificado e pela eletroforese em gel de poliacrilamida - dodecil sulfato de sódio (SDS-PAGE), bem como para avaliação citológica. Por meio de venopunção, coletou-se sangue para determinação da concentração de proteínas séricas. Treze olhos (50% das amostras) apresentaram valor proteico médio de 40,3 mg/dL±6,45 e a eletroforese demonstrou presença de proteínas de massas mais elevadas que 43 KDa. Houve ausência de células em 96,15% das amostras (25 olhos). Equinos hígidos apresentaram baixa concentração de proteínas no HA. Já a correlação entre proteína no humor aquoso/proteína plasmática total foi de 0,56%.(AU)

Evaluation of equine aqueous humor (AH) proteins can help the diagnosis of uveitis. However, studies on proteins in this fluid in healthy horses are scarce and present variations according to the methodology employed. This study aimed to perform protein analysis and cytology of equine aqueous humor of healthy horses and verify its correlation with plasmatic proteins. Thirteen adult horses (26 eyes), mixed breed, male or female were evaluated. A volume of 0.5 mL of aqueous humor was collected through aqueocentesis from both eyes. The samples were submitted to protein quantification by modified Bradford method and to sodium dodecyl sulphate - polyacrylamide gel electrophoresis (SDS-PAGE), and to cytological evaluation. Blood was collected for determination of plasmatic protein concentration. Thirteen eyes (50% of the samples) had values larger than zero by the Bradford method, with an average of 40.3 mg/dl±6.45. Electrophoresis showed presence of higher masses of proteins (43 KDa). There were no cells in 96.15% of the samples (25 eyes). Healthy equines presented low protein concentration in the HA. The ratio between protein concentration in the aqueous humor / total plasma protein of 0.56%.(AU)