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.

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)

Neutrophil proteomic analysis reveals the participation of antioxidant enzymes, motility and ribosomal proteins in the prevention of ischemic effects by preconditioning.

Intestinal ischemia and reperfusion injury are widely used models, which result into tissue injury and multiple organ failure also observed after trauma and surgery. Ischemic preconditioning (IPC) preceding ischemia and reperfusion (IR) was shown to attenuate this injury and has a potential therapeutic application; however the exact underlying mechanism is not clear. Neutrophils play an important role in the mechanism of injuries caused by ischemia and reperfusion while IPC led to a decrease in neutrophil stimulation and activation. The effect of preconditioning on the neutrophil proteome is unclear. Proteomic analysis has been ratified as an appropriate tool for studying complex systems. In order to evaluate the effect of IPC preceding 45min of ischemia on the proteome of neutrophils we used Wistar rats divided in four experimental groups: Control, sham laparotomy, intestinal ischemia reperfusion and ischemic preconditioning. After neutrophil separation, proteins were extracted, trypsin digested and the resulting peptides were iTRAQ labeled followed by HILIC fractionation and nLC-MS/MS analysis. After database searches, normalization and statistical analysis our proteomic analysis resulted in the identification of 2437 protein groups that were assigned to five different clusters based on the relative abundance profiles among the experimental groups. The clustering followed by statistical analysis led to the identification of significantly up and downregulated proteins in IR and IPC. Cluster based KEGG pathways analysis revealed up- regulation of actin cytoskeleton, metabolism, Fc gamma R mediated phagocytosis, chemokine signaling, focal adhesion and leukocyte transendothelial migration whereas downregulation in ribosome, spliceosome, RNA transport, protein processing in endoplasmic reticulum and proteasome, after intestinal ischemic preconditioning. Furthermore, enzyme prediction analysis revealed the regulation of some important antioxidant enzymes and having their role in reactive oxygen species production. To our knowledge, this work describes the most comprehensive and detailed quantitative proteomic study of the neutrophil showing the beneficial role of ischemic preconditioning and its effects on the neutrophil proteome. This data will be helpful to understand the effect of underlying protective mechanisms modulating the role of PMNs after IPC and provide a trustworthy basis for future studies. BIOLOGICAL SIGNIFICANCE: Preconditioning is a relevant strategy to overcome clinical implications from ischemia and reperfusion. Such implications have the neutrophil as a major player. Although many publications describe specific biochemical and physiological roles of the neutrophil in such conditions, there is no report of a proteomic study providing a broader view of this scenario. Here we describe a group of proteins significantly regulated by ischemia and reperfusion being such regulation prevented by preconditioning. Such finding may provide relevant information for a deeper understanding of the mechanisms involved, as well as serve as basis for future biomarker or drug target assays.

Anticancer peptides and proteins: a panoramic view.

Despite progress in understanding carcinogenesis, cancer is still a major cause of morbidity and mortality worldwide. Treatment is complicated by the resistance to chemotherapeutics acquired by tumors, ability to repair damaged DNA, alteration of the interaction between drug and its target, and defects in the machinery that leads to apoptosis. The development of novel therapies based on biologically active peptides, especially antimicrobial peptides (AMPs), and proteins has emerged as a new strategy to defeat cancer.

Proteome analysis of Nelore bull (Bos taurus indicus) seminal plasma.

The Nelore bull (Bos taurus indicus) seminal plasma proteome was analyzed by MALDI-TOF MS and two-dimensional gel electrophoresis. A total of 260 spots were visualized in the 2-DE gel (pI range 3-10) and 13 spots could be identified by peptide mass fingerprinting corresponding to 11 different polypeptides. The results allowed the creation of the first proteomic map of Bos taurus indicus seminal plasma. The roles of the identified proteins in the bull seminal plasma are discussed.

Percutaneous catheterization of the subclavian vein in hemophiliac patients: report of 47 cases.

Hemophiliac patients frequently require venous access, but usually their peripheral veins are collapsed and not puncturable. Several procedures employed to overcome this difficulty present a high complication rate. The authors report 47 cases of percutaneous subclavian vein catheterization (PSC) in hemophiliacs with an overall complication rate of 23%. Six patients (13%) had complications attributed to the coagulation disorder. The complications attributed to the coagulopathy and presenting major clinical importance occurred early in the series. After the improvement of the coagulation control with a routine AHG administration policy, the authors observed that among the last 22 patients there were 3 complications related to the coagulopathy, all of them without major clinical importance. The authors conclude that, in the lack of a safer access, PSC seems to be an acceptable procedure in hemophiliac patients, if employed with strict indication, accurate technique and rigorous control of the coagulopathhy.

[Syphilitic aneurysm communicating with an aortic sinus of Valsalva. A case report]. / Aneurisma sifilítico comunicando-se com um seio de Valsalva aórtico. Relato de caso.

The authors present the case of a 27-year old woman with an aneurysm, possibly originating from an ectopic coronary sprout and communicating with the right anterior sinus of Valsalva. Secondary syphilitic lesions were observed. By virtue of its great size and localization, this aneurysm produced obstruction of the outlet of the right ventricle and dislocation of the left coronary artery. The authors conclude that this aneurysm was a congenital anomaly because of its great volume, configuration, the way it opened in the aortic right anterior sinus of Valsalva, the normal aortic wall and valve, and normal sinuses of Valsalva, observed at surgery. The follow-up was uneventful.