A peptide fraction from hardened common beans (Phaseolus vulgaris) induces endothelium-dependent antihypertensive and renal effects in rats.

Beans reached the research spotlight as a source of bioactive compounds capable of modulating different functions. Recently, we reported antioxidant and oxidonitrergic effect of a low molecular weight peptide fraction (<3 kDa) from hardened bean (Phaseolus vulgaris) in vitro and ex vivo, which necessitate further in vivo assessments. This work aimed to evaluate the hypotensive effect and the involved physiological mechanisms of the hardened common bean peptide (Phaseolus vulgaris) in normotensive (Wistar) and hypertensive (SHR) animals. Bean flour was combined with a solution containing acetonitrile, water and formic acid (25: 24: 1). Protein extract (PV3) was fractioned (3 kDa membrane). We assessed PV3 effects on renal function and hemodynamics of wistar (WT-normotensive) and spontaneously hypertensive rats (SHR) and measured systemic arterial pressure and flow in aortic and renal beds. The potential endothelial and oxidonitrergic involvements were tested in isolated renal artery rings. As results, we found that PV3: I) decreased food consumption in SHR, increased water intake and urinary volume in WT, increased glomerular filtration rate in WT and SHR, caused natriuresis in SHR; II) caused NO- and endothelium-dependent vasorelaxation in renal artery rings; III) reduced arterial pressure and resistance in aortic and renal vascular beds; IV) caused antihypertensive effects in a dose-dependent manner. Current findings support PV3 as a source of bioactive peptides and raise the potential of composing nutraceutical formulations to treat renal and cardiovascular diseases.

Polysaccharide-Based Membrane Biocompatibility Study of Anacardium occidentale L. and Polyvinyl Alcohol after Subcutaneous Implant in Rats.

Polymeric membranes are a viable and sustainable option for the biotechnology industry from an economic and environmental point of view. In this study, we evaluated tissue response and tolerance to the implantation of a polymeric membrane prepared with cashew gum polysaccharide (CGP) associated with polyvinyl alcohol (PVA). The objective was to characterize the biocompatibility of the CGP/PVA membrane in vivo. Following the evaluation criteria of the ISO 10993-6 standard, we demonstrated that the CGP/PVA membrane showed moderate tissue reaction, with a non-irritating ISO pattern, a thinner fibrous capsule, and a smaller amount of collagen compared to the positive control group. At 30 and 60 days, the membrane presented a similar amount of mast cells to that observed in the negative control group. The data demonstrate that the CGP/PVA membrane presents biocompatibility in accordance with the ISO 10993-6 standard.

Larvicidal potential of cell wall degrading enzymes from Trichoderma asperellum against Aedes aegypti (Diptera: Culicidae).

Aedes aegypti is a mosquito vector of arboviruses such as dengue, chikungunya, zika and yellow fever that cause important public health diseases. The incidence and gravity of these diseases justifies the search for effective measures to reduce the presence of this vector in the environment. Bioinsecticides are an effective alternative method for insect control, with added ecological benefits such as biodegradability. The current study demonstrates that a chitinolytic enzyme complex produced by the fungus Trichoderma asperellum can disrupt cuticle formation in the L3 larvae phase of A. aegypti, suggesting such biolarvicidal action could be used for mosquito control. T. asperellum was exposed to chitin from different sources. This induction of cell wall degrading enzymes, including chitinase, N-acetylglucosaminidase and ß-1,3-glucanase. Groups of 20 L3 larvae of A. aegypti were exposed to varying concentrations of chitinolytic enzymes induced with commercial chitin (CWDE) and larvae cell wall degrading enzymes (L-CWDE). After 72 h of exposure to the CWDE, 100% of larvae were killed. The same percent mortality was observed after 48 h of exposure to L-CWDE at half the CWDE enzyme mixture concentration. Exoskeleton deterioration was further observed by scanning and electron microscopy. Our findings indicate that L-CWDE produced by T. asperellum reflect chitinolytic enzymes with greater specificity for L3 larval biomolecules. This specificity is characterized by the high percentage of mortality compared with CWDE treatments and also by abrupt changes in patterns of the cellular structures visualized by scanning and transmission electron microscopy. These mixtures of chitinolytic enzymes could be candidates, as adjuvant or synergistic molecules, to replace conventional chemical insecticides currently in use.

Cashew Gum Polysaccharide Nanoparticles Grafted with Polypropylene Glycol as Carriers for Diclofenac Sodium.

This investigation focuses on the development and optimization of cashew gum polysaccharide (CGP) nanoparticles grafted with polypropylene glycol (PPG) as carriers for diclofenac sodium. The optimization of parameters affecting nanoparticles formulation was performed using a central composite rotatable design (CCRD). It was demonstrated that the best formulation was achieved when 10 mg of CGP was mixed with 10 µL of PPG and homogenized at 22,000 rpm for 15 min. The physicochemical characterization evidenced that diclofenac was efficiently entrapped, as increases in the thermal stability of the drug were observed. The CGP-PPG@diclofenac nanoparticles showed a globular shape, with smooth surfaces, a hydrodynamic diameter around 275 nm, a polydispersity index (PDI) of 0.342, and a zeta potential of -5.98 mV. The kinetic studies evidenced that diclofenac release followed an anomalous transport mechanism, with a sustained release up to 68 h. These results indicated that CGP-PPG nanoparticles are an effective material for the loading/release of drugs with similar structures to diclofenac sodium.

Chemical characterization and pharmacological assessment of polysaccharide free, standardized cashew gum extract (Anacardium occidentale L.).

ETHNOPHARMACOLOGICAL RELEVANCE: The cashew gum (Anacardium occidentale L.) is used in traditional Brazilian medicine in the treatment of inflammatory conditions, asthma, diabetes, and gastrointestinal disturbances. AIM OF THE STUDY: In the present study, we aimed at forming a chemical characterization and investigation of the antinociceptive and anti-inflammatory activities of the aqueous extract of cashew gum without the presence of polysaccharides in its composition (CGE). MATERIALS AND METHODS: The CGE was obtained after the precipitation and removal of polysaccharides through the use of acetone. After, the acetone was removed by rotaevaporation, and the concentrated extract was lyophilized. The chemical characterization of CGE was performed by liquid chromatography mass spectrometry (LC-MS) and tandem mass spectrometry (MS/MS) analyses. Mice were used for the evaluation of the antinociceptive and anti-inflammatory activities. CGE was analyzed via the Irwin test, acetic acid-induced writhing test, formalin-induced pain test, and carrageenan-induced paw edema test. The motor activity or probable sedation was verified through the chimney, open-field, and sodium pentobarbital-induced sleep tests. We investigated if the analgesic and anti-inflammatory effects of CGE depend of reduction in PGE2 levels, were performed the carrageenan or PGE2-induced hyperalgesia tests. RESULTS: The chemical characterization of CGE showed the presence of anacardic acids as the predominant phytoconstituents. The treatment with CGE (75, 150, and 300mg/kg, p.o.) inhibited the number of writhing in a dose-dependent manner. With an intermediate dose, CGE did not cause motor impairment with the chimney test or alterations in either the open-field or sodium pentobarbital-induced sleep. In the formalin-induced pain test, CGE (150mg/kg, p.o.) produced an antinociceptive effect only in the first phase of the test, suggesting anti-inflammatory activity. With the same dosage, CGE also reduced the carrageenan-induced paw edema at all hours of the test, confirming its anti-inflammatory effect. Furthermore, CGE (150mg/kg, p.o.) presented an antihyperalgic effect at all hours of the carrageenan-induced hyperalgesia test. However, this dose of CGE was not able to reduce the hyperalgesia induced by PGE2, suggesting that the anti-inflammatory effect of this extract depends on the reduction in the PGE2 levels. CONCLUSION: The anacardic acids are the predominant phytoconstituents identified in the CGE. The action mechanisms of CGE suggest the reduction in the PGE2 levels. These findings support the use of cashew gum in popular medicine and demonstrate that part of its antinociceptive and anti-inflammatory effects should also be attributed to the presence of anacardic acids in its composition, independent of the presence of polysaccharides.

Mucoadhesive Properties of Thiolated Pectin-Based Pellets Prepared by Extrusion-Spheronization Technique.

The aim of this study was to develop mucoadhesive pellets on a thiolated pectin base using the extrusion-spheronization technique. Thiolation of pectin was performed by esterification with thioglycolic acid. The molecular weight and thiol group content of the pectins were determined. Pellets containing pectin, microcrystalline cellulose, and ketoprofen were prepared and their mucoadhesive properties were evaluated through a wash-off test using porcine intestinal mucosa. The in vitro ketoprofen release was also evaluated. Thiolated pectin presented a thiol group content of 0.69 mmol/g. Thiolation caused a 13% increase in polymer molecular weight. Pellets containing thiolated pectin were still adhering to the intestinal mucosa after 480 min and showed a more gradual release of ketoprofen. Conversely, pellets prepared with nonthiolated pectin showed rapid disintegration and detached after only 15 min. It can be concluded that thiolated pectin-based pellets can be considered a potential platform for the development of mucoadhesive drug delivery systems for the oral route.

A Review of the Latest Advances in Encrypted Bioactive Peptides from Protein-Rich Waste.

Bioactive peptides are considered the new generation of biologically active regulators that not only prevent the mechanism of oxidation and microbial degradation in foods but also enhanced the treatment of various diseases and disorders, thus increasing quality of life. This review article emphasizes recent advances in bioactive peptide technology, such as: (i) new strategies for transforming bioactive peptides from residual waste into added-value products; (ii) nanotechnology for the encapsulation, protection and release of controlled peptides; and (iii) use of techniques of large-scale recovery and purification of peptides aiming at future applications to pharmaceutical and food industries.

Lipase entrapment in PVA/Chitosan biodegradable film for reactor coatings.

This study reports the development and characterization of novel biodegradable film, based on chitosan and polyvinyl alcohol containing lipase entrapped. The films showed a thickness of 70.4 and 79 µm to PVA/Chitosan and PVA/Chitosan/Lipase, respectively. The entrapment of lipase in PVA/Chitosan film resulted in increasing of 69.4% tensile strength (TS), and 52.4% of elongation. SEM images showed the formation of a continuous film, without pores or cracks. The lipase entrapment efficiency was estimated in 92% and the films were repeatedly used for 25 hydrolytic cycles, maintaining 62% of initial activity. The PVA/Chitosan/Lipase film was used for olive oil hydrolysis of high performance. These results indicate that PVA/Chitosan/Lipase is a promising material for biotechnology applications such as triacylglycerol hydrolysis and biodiesel production.

Immobilized horseradish peroxidase on discs of polyvinyl alcohol-glutaraldehyde coated with polyaniline.

Discs of network polyvinyl alcohol-glutaraldehyde (PVAG) were synthesized and coated with polyaniline (PANI) using glutaraldehyde as a chemical arm (PVAG-PANIG-HRP disc). The best conditions for the immobilization were established as about 1.0 mg mL(-1) of protein, for 60 min and pH 5.5. The soluble enzyme lost all of its activity after incubation at 70°C for 15 min, whereas the PVAG-PANIG-HRP disc retained about half of the initial activity for pyrogallol. The same PVAG-PANIG-HRP disc was used consecutively three times without any activity lossbut presented 25% of the initial activity after the 7th use. PVAG-PANIG-HRP disc retained approximately 80% and 60% of its initial activity after 60 and 80 days of storage, respectively. Resorcinol, m-cresol, catechol, pyrogallol, α-naphthol, ßnaphthol, and 4, 4'-diaminodiphenyl benzidine were efficiently oxidized by the PVAG-PANIG-HRP disc (from about 70% to 90%), and it was less efficient towards aniline, phenol, and 2-nitrosonaphthol.

Immobilization of peroxidase onto magnetite modified polyaniline.

The present study describes the immobilization of horseradish peroxidase (HRP) on magnetite-modified polyaniline (PANImG) activated with glutaraldehyde. After the optimization of the methodology, the immobilization of HRP on PANImG produced the same yield (25%) obtained for PANIG with an efficiency of 100% (active protein). The optimum pH for immobilization was displaced by the effect of the partition of protons produced in the microenvironment by the magnetite. The tests of repeated use have shown that PANImG-HRP can be used for 13 cycles with maintenance of 50% of the initial activity.

Optimal Conditions for Continuous Immobilization of Pseudozyma hubeiensis (Strain HB85A) Lipase by Adsorption in a Packed-Bed Reactor by Response Surface Methodology.

This study aimed to develop an optimal continuous process for lipase immobilization in a bed reactor in order to investigate the possibility of large-scale production. An extracellular lipase of Pseudozyma hubeiensis (strain HB85A) was immobilized by adsorption onto a polystyrene-divinylbenzene support. Furthermore, response surface methodology (RSM) was employed to optimize enzyme immobilization and evaluate the optimum temperature and pH for free and immobilized enzyme. The optimal immobilization conditions observed were 150 min incubation time, pH 4.76, and an enzyme/support ratio of 1282 U/g support. Optimal activity temperature for free and immobilized enzyme was found to be 68°C and 52°C, respectively. Optimal activity pH for free and immobilized lipase was pH 4.6 and 6.0, respectively. Lipase immobilization resulted in improved enzyme stability in the presence of nonionic detergents, at high temperatures, at acidic and neutral pH, and at high concentrations of organic solvents such as 2-propanol, methanol, and acetone.

Composição química e antinutrientes presentes nas amêndoas cruas e torradas de chicha (Sterculia striata A. St. Hill & Naudin) / Chemical composition and antinutrients of raw and roasted chicha almonds (Sterculia striata A. St. Hill & Naudin)

OBJETIVO: Analisar os componentes químicos e antinutrientes antes e após torrefação da amêndoa de chichá da espécie Sterculia striata A. St. Hill & Naudin, oriunda de Corrente, Piauí. MÉTODOS: Foram realizadas análises: centesimal, perfil de ácidos graxos, antinutrientes (taninos, inibidor de tripsina, inibidor de amilase, lectina, fitatos), compostos fenólicos e atividade de peroxidase e polifenoloxidase. RESULTADOS: Os resultados da análise centesimal da amêndoa crua e torrada foram: lipídeos de 25,1 por cento e 26,2 por cento, carboidratos de 44,4 por cento e 45,6 por cento, proteína de 20,8 por cento e 22,1 por cento, cinzas de 3,7 por cento e 4,0 por cento, fibra alimentar total de 12,3 por cento e 10,4 por cento, respectivamente. Nas amêndoas cruas e torradas não foram encontradas lectinas, taninos, inibidores de tripsina e alfa-amilase. Verificou-se a ausência de peroxidase e polifenoloxidase e conteúdo de compostos fenólicos de 107,7mg/100g e 108,9mg/100g para amêndoas cruas e torradas, respectivamente. CONCLUSÃO: A torrefação realizada a 205ºC por 11 minutos diminuiu o teor de fitatos de 10,6mg/g para 5,5mg/g. Por fim, as amêndoas de chichá, cruas e torradas, apresentam alto teor de proteínas, fibras, ácidos graxos monoinsaturados e saturados.

OBJECTIVE: This study analyzed the chemical components and antinutrients present in raw and roasted chichá almonds from the species Sterculia striata A. St. Hill & Naudin harvested in Corrente, Piauí. METHODS: The following were determined: percent composition, fatty acid profile, antinutrients (tannins, trypsin inhibitor, amylase inhibitor, lectin, phytates), phenolic compounds and peroxidase and polyphenol oxidase activities. RESULTS: The percentage composition of the raw and lipid almonds were respectively: 25.1 percent and 26.2 percent fats, 44.4 percent and 45.6 percent carbohydrates, 20.8 percent and 22.1 percent protein, 3.7 percent and 4.0 percent ashes, and 12.3 percent and 10.4 percent total fiber. Neither raw nor roasted almonds contained lectins, tannins, trypsin inhibitors, alpha-amylase, peroxidase and polyphenol oxidase. Contents of phenolic compounds were 107.7mg/100g and 108.9mg/100 for the raw and roasted almonds, respectively. CONCLUSION: Roasting at 205ºC for 11 minutes reduced phytate content from 10.6mg/g to 5.5mg/g. Raw and roasted chichá almonds have a high content of proteins, fibers and monounsaturated and saturated fatty acids.

Purification of Paracoccidioides brasiliensis catalase P: subsequent kinetic and stability studies.

Catalases are essential components of the cellular equipment to cope with oxidative stress. Here we have purified a highly abundant catalase P of Paracoccidioides brasiliensis (PbCatP) that is preferentially expressed in the parasitic yeast phase. This oxidative stress-induced protein was isolated from yeast cells grown in the presence of 15 mM of hydrogen peroxide (H(2)O(2)). We have used consecutive steps of protein precipitation and gel filtration chromatography to achieve the purified protein. Protein purification was validated using matrix-assisted laser desorption ionization time-of-flight mass spectrometry and bioinformatics analysis. The purified enzyme showed strong similarity to small-subunit catalases. Like most monofunctional catalases, PbCatP is a homotetramer, resistant to inactivation by acidic conditions, temperature and denaturants. Furthermore, the kinetic behaviour of catalase P was observed to be different at low compared to high H(2)O(2) concentrations. The results demonstrated that a purified PbCatP is a homotetrameric enzyme, classified as a small subunit catalase.

Biochemical characterization of selected plant species from Brazilian Savannas

O Cerrado é constituído por inúmeras espécies vegetais com potencial econômico, as quais são utilizadas para os mais variados fins, como medicinal e nutricional. O objetivo deste trabalho foi analisar e quantificar a presença de atividade enzimática de peroxidases e proteases e fatores antinutricionais, como lectinas e inibidores de proteases, além de polifenóis e taninos em algumas espécies nativas do Cerrado. O material vegetal utilizado foram sementes de Annona crassiflora Mart. (araticum), Hymenaea courbaril L. var. courbaril (jatobá), Plathymenia reticulata Benth. (vinhático), Zanthoxylum rhoifolium Lam. (maminha de porca), Apeiba tibourbou Aubl. (pau jangada), Salacia crassiflora (Mart.) G. Don. (bacupari) e Sclerolobium paniculatum Vog. (carvoeiro), coletadas na cidade de Goiânia e municípios de Jataí e Caldas Novas, estado de Goiás. O uso potencial destas plantas e suas enzimas na indústria de alimentos, poderia resultar em aplicações ao aparecimento de novos produtos a partir das matérias-primas tradicionais, além do uso de novas fontes de alimentos.