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The emergence of antibiotic resistance poses a serious and challenging threat to healthcare systems, making it imperative to discover novel therapeutic options. This work reports the isolation and characterization of a thermostable trypsin inhibitor from chia (Salvia hispanica L.) seeds, with antibacterial activity against Staphylococcus aureus sensitive and resistant to methicillin. The trypsin inhibitor ShTI was purified from chia seeds through crude extract heat treatment, followed by affinity and reversed-phase chromatography. Tricine-SDS-PAGE revealed a single glycoprotein band of ~ 11 kDa under nonreducing conditions, confirmed by mass spectrometry analysis (11.558 kDa). ShTI was remarkably stable under high temperatures (100 °C; 120 min) and a broad pH range (2-10; 30 min). Upon exposure to DTT (0.1 M; 120 min), ShTI antitrypsin activity was partially lost (~ 38%), indicating the participation of disulfide bridges in its structure. ShTI is a competitive inhibitor (Ki = 1.79 × 10-8 M; IC50 = 1.74 × 10-8 M) that forms a 1:1 stoichiometry ratio for the ShTI:trypsin complex. ShTI displayed antibacterial activity alone (MICs range from 15.83 to 19.03 µM) and in combination with oxacillin (FICI range from 0.20 to 0.33) against strains of S. aureus, including methicillin-resistant strains. Overproduction of reactive oxygen species and plasma membrane pore formation are involved in the antibacterial action mode of ShTI. Overall, ShTI represents a novel candidate for use as a therapeutic agent for the bacterial management of S. aureus infections.

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Seed legumes have played a major role as a crop worldwide, being cultivated on about 12% to 15% of Earth's arable land; nevertheless, their use is limited by, among other things, the presence of several antinutritional factors (ANFs - naturally occurring metabolites that the plant produces to protect itself from pest attacks.) Trypsin inhibitors (TIs) are one of the most relevant ANFs because they reduce digestion and absorption of dietary proteins. Several methods have been developed in order to inactivate TIs, and of these, thermal treatments are the most commonly used. They cause loss of nutrients, affect functional properties, and require high amounts of energy. Given the above, new processes have emerged to improve the nutritional quality of legumes while trying to solve the problems caused by the use of thermal treatments. This review examines and discusses the methods developed by researchers to inactivate TI present in legumes and their effects over nutritional and functional properties.

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We have prepared a review of the physical-chemical composition and the functional and anti-nutritional properties of quinoa (Chenopodium quinoa Willd.). It is a plant of the Chenopodiaceae family, originally from the Andean regions, adaptable to different types of soils and climatic conditions. Its composition has attracted the attention of scientific community for its high nutritional value, being rich in proteins, lipids, fibers, vitamins, and minerals, with an extraordinary balance of essential amino acids. It is also gluten-free, a characteristic that enables its use by celiac patients. In spite of all these attributes, quinoa is not widely used by consumers due to the high cost of imported grain and little knowledge of its benefits. More studies are required to increase knowledge about this "pseudo-cereal" to demonstrate its functional and nutritional benefits and to study its anti-nutritional effects, since it presents high commercial value and excellent nutritional quality.

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Proteasome inhibitors have been described as an important target for cancer therapy due to their potential to regulate the ubiquitin-proteasome system in the degradation pathway of cellular proteins. Here, we reported the effects of a Bowman-Birk-type protease inhibitor, the Black-eyed pea Trypsin/Chymotrypsin Inhibitor (BTCI), on proteasome 20S in MCF-7 breast cancer cells and on catalytic activity of the purified 20S proteasome from horse erythrocytes, as well as the structural analysis of the BTCI-20S proteasome complex. In vitro experiments and confocal microscopy showed that BTCI readily crosses the membrane of the breast cancer cells and co-localizes with the proteasome in cytoplasm and mainly in nucleus. Indeed, as indicated by dynamic light scattering, BTCI and 20S proteasome form a stable complex at temperatures up to 55°C and at neutral and alkaline pHs. In complexed form, BTCI strongly inhibits the proteolytic chymotrypsin-, trypsin- and caspase-like activities of 20S proteasome, indicated by inhibition constants of 10(-7) M magnitude order. Besides other mechanisms, this feature can be associated with previously reported cytostatic and cytotoxic effects of BTCI in MCF-7 breast cancer cells by means of apoptosis.

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BACKGROUND: Jatropha cordata and Jatropha cardiophylla are native to northwestern Mexico and are adapted to arid and semi-arid conditions (<500 mm of precipitation and temperatures from 8 to 45 °C). The aim of this study was to evaluate the chemical composition of J. cordata and J. cardiophylla kernels and oils as well as antinutrients in the defatted kernel meals of these species. RESULTS: Kernels of J. cordata and J. cardiophylla seeds analysed in this study were rich in crude protein (283 and 289 g kg(-1) respectively) and lipid (517 and 537 g kg(-1) respectively). The main fatty acids in J. cordata and J. cardiophylla oils were linoleic and oleic acids. High levels of trypsin inhibitor and phytates and low levels of saponins were present in the meals. The phorbol ester contents in J. cordata and J. cardiophylla kernel meals were 2.73 and 1.46 mg g(-1) respectively. CONCLUSION: For both J. cordata and J. cardiophylla it could be inferred that (a) the oil and kernel meal were toxic and the kernel meal could be used as livestock feed only after detoxification, (b) the oil could be used for non-alimentary purposes, i.e. biodiesel production, and (c) the seed or oil could be used for isolating various bioactive compounds for pharmaceutical and agricultural applications.

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Aedes aegypti control is crucial to reducing dengue fever. Aedes aegypti larvae have developed resistance to organophosporous insecticides and the use of natural larvicides may help manage larval resistance by increasing elements in insecticide rotation programs. Here, we report on larvicidal activity of Moringa oleifera flower extract against A. aegypti L(1), L(2), L(3), and L(4) as well as the effect of flower extract on gut trypsin and whole-larval acetylcholinesterase from L(4.) In addition, the heated flower extract was investigated for larvicidal activity against L(4) and effect on larval gut trypsin. Moringa oleifera flower extract contains a proteinaceous trypsin inhibitor (M. oleifera flower trypsin inhibitor, MoFTI), triterpene (ß-amyrin), sterol (ß-sitosterol) as well as flavonoids (kaempferol and quercetin). Larvicidal activity was detected against L(2), L(3), and L(4) (LC(50) of 1.72%, 1.67%, and 0.92%, respectively). Flower extract inhibited L(4) gut trypsin (MoFTI K(i) = 0.6 nM) and did not affect acetylcholinesterase activity. In vivo assay showed that gut trypsin activity from L(4) treated with M. oleifera flower extract decreased over time (0-1,440 min) and was strongly inhibited (98.6%) after 310 min incubation; acetylcholinesterase activity was not affected. Thermal treatment resulted in a loss of trypsin inhibitor and larvicidal activities, supporting the hypothesis that flower extract contains a proteinaceous trypsin inhibitor that may be responsible for the deleterious effects on larval mortality.

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The biochemical and functional properties of 2 hard-to-cook common bean cultivars (Phaseolus vulgaris, L.) were investigated after the extrusion process. Beans of BRS pontal and BRS grafite cultivars were milled and extruded at 150 degrees C, with a compression ratio screw of 3 : 1, 5-mm die, and screw speed of 150 rpm. Extrudate flours were evaluated for water solubility (WS), water absorption index (WAI), oil absorption capacity (OAC), foaming capacity (FC), emulsifying activity (EA), antinutritional factors, and in vitro protein and starch digestibility. Results indicated that the extrusion significantly decreased antinutrients such as phytic acid, lectin, alpha-amylase, and trypsin inhibitors, reduced the emulsifying capacity and eliminated the FC in both BRS pontal and BRS grafite cultivars. In addition, the WS, WAI, and in vitro protein and starch digestibility were improved by the extrusion process. These results indicate that it is possible to produce new extruded products with good functional and biochemical properties from these common bean cultivars.

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Jabuticaba, Myrciaria cauliflora Berg, is a native Brazilian fruit; very little is known about the chemistry of its components, specially the bioactives compounds. The purpose of this paper was to determine the centesimal composition and some bioactives compounds of the whole fruit and fractions of two varieties of jabuticaba (Paulista and Sabará). The experiment was conducted in a completely randomized design on a 4 x 2 x 3 factorial scheme (4 factors: whole fruit, skin, pulp and seed; 2 varieties: Paulista e Sabará, and 3 replicates). The fruits were sampled, selected, weighted, sanitized and fractionated into whole fruit, skin, pulp and seed that were grinded, frozen and lyophilized to a constant weight. Protein contents and of ether extract were low for all fractions. The ash contents indicate elevated levels of minerals, specially on the Sabará variety. The content of alimentary fibers differed widely among fractions, with no differences between varieties. The skin fractions presented the highest levels of fiber, 33.80 g/100 g on Paulista and 33.23 g/100 g on Sabará, most of it was insoluble fibers. The greatest amount of non nitrogen extract was found in the pulp fraction, while the lowest was found in the skin, with no differences between varieties. Regarding bioactives compounds, only polyphenols were presented at considerable levels in all the fractions but the pulp one, showing the need for better characterization before using the fruit in the food industry.

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Nutrient and antinutritional/toxic factors present in some edible flowers consumed in Mexico were determined. The edible flowers were: Agave salmiana, Aloe vera, Arbutus xalapensis, Cucurbita pepo (cultivated), Erythrina americana, Erythrina caribaea, Euphorbia radians benth and Yucca filifera. The nutrient content in the flowers studied is similar to that of the edible leaves and flowers studied mainly in Africa. The moisture content of the flowers varied from 860 to 932 g kg(-1). Crude protein (CP) was between 113 to 275 g kg(-1) DM, crude fiber, 104 to 177 g kg(-1) DM and the nitrogen free extract, between 425 to 667 g kg(-1) DM. The highest chemical score (CS) was found in E. americana and A. salmiana; in five samples the limiting amino acid was lysine, and in three of them it was tryptophan. Trypsin inhibitors and hemaglutinnins had a very low concentration. Alkaloids were present in both the Erythrina species and the saponins in A. salmiana and Y. filifera. Cyanogenic glucosides were not found in the studied flowers. The traditional process of preparing these specific flowers before consumption is by cooking them and discarding the broth; in this way the toxic substances are diminished or eliminated. These edible flowers from wild plants consumed in local areas of the country play an important role in the diet of the people at least during the short time of the season where they are blooming.

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Pigeon pea (Cajanus cajan var. aroíto) seeds were fermented in order to remove antinutritional factors and to obtain functional legume flour to be used as pasta ingredients. Fermentation brought about a drastic reduction of alpha-galactosides (82%), phytic acid (48%), and trypsin inhibitor activity (39%). Fermented legume flours presented a notable increase of fat and total soluble available carbohydrates, a slight decrease of protein, dietary fiber, calcium, vitamin B2, vitamin E, and total antioxidant capacity, and a decrease of soluble dietary fiber, Na, K, Mg, and Zn contents. No changes were observed in the level of starch and tannins as a consequence of fermentation. The fermented flour was used as an ingredient to make pasta products in a proportion of 5, 10, and 12%. The supplemented pasta products obtained had longer cooking times, higher cooking water absorptions, higher cooking loss, and higher protein loss in water than control pasta (100% semolina). From sensory evaluations, fortified pasta with 5 and 10% fermented pigeon pea flour had an acceptability score similar to control pasta. Pasta supplemented with 10% fermented pigeon pea flour presented higher levels of protein, fat, dietary fiber, mineral, vitamin E, and Trolox equivalent antioxidant capacity than 100% semolina pasta and similar vitamins B1 and B2 contents. Protein efficiency ratios and true protein digestibility improved (73 and 6%, respectively) after supplementation with 10% fermented pigeon pea flour; therefore, the nutritional value was enhanced.

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Trypsin inhibitors (TI), tannins, and lectins appear to have a role in preventing chronic diseases in humans. The genetic variability of these traits in common bean needs to be ascertained in order to increase levels through breeding. The variability of TI, tannin, and lectins was determined in five bean cultivars grown at five locations in Mexico. TI and tannins contents in colored beans that belong to the Jalisco race were higher (11.1-11.9 trypsin units inhibited (TUI)/mg and 29.0-38.1 mg catechin equivalent (CE)/g, respectively) than cultivars of the Durango race (7.9-8.3 TUI/mg and 16.8-19.9 CE/mg, respectively). Bayo Victoria, a Durango race cultivar, had three times more lectins than levels reported for soybean. Cultivar influenced TI and tannins contents (p < 0.001), whereas site affected lectins (p < 0.001). An increase in levels of TI and tannins could be enhanced through breeding.

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The soybean cultivar BR 36 with conventional levels of trypsin inhibitors activity and the soybean line BRM95-5262, which was genetically selected to contain low activity of trypsin inhibitors were used for biological assays with rats. BR 36 and BRM95-5262 contained 40 and 20, and 30 and 20% of relative residual activity of trypsin inhibitors, respectively. The mean values of PER and NPR showed that treatments with crude soybeans were minor than treatments with soybean thermically processed. However, the treatments of thermically processed soybean did not showed significative differences (p > or = 0.05). When the trypsin inhibitors activity were 8.61 and 8.44 UIT/mg of samples or 20 and 30% of relative residual activity of cultivar BR 36 and line BRM95-5262, respectively, it was observed that mean values of PER and NPR were not significatives. The mean values of CDA and CDV of treatments with crude soybeans were minor than treatment with casein and similar to the treatments with soybean thermically processed. So, it can be concluded that the biological evaluation obtained with soybean protein were dependent of initial trypsin inhibitors activities and of its respective thermical treatment. There was advantage in the use of BRM95-5262 soybean line with low trypsin inhibitors activity.

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High values of residual trypsin inhibitory activity found for heat-treated beans indicated the occurrence of artifices due to the methodology. A critical evaluation of the methods in use and of possible interferents was performed trying to determine the most adequate way of measuring residual trypsin inhibitory activity. Results showed that extraction conditions, pH of reaction and sample preparation are responsible for significant alterations of the values found, requiring careful standardization. This is the first report about the influence of lyophilization upon residual trypsin inhibitory activity of cooked beans.

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Seeds of soybean cultivar BR 16 were submitted to malting for 24 and 48 hours and to extrusion originating flakes used as ingredient to make biscuits which were chemically and sensorially evaluated. Macronutrient composition did not show any relevant modification presenting only a small decrease in carbohydrates and increase in the relative lipid composition. Some anti-nutritional factors were affected by the processes. Trypsin inhibitors decreased about 50% after 24 h of malting and disappeared after extrusion. Phytic acid was resistant to the processes while alfa-galactosides presented about 80% of loss as a result of the combination of malting and extrusion.

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Sementes de soja cultivar BR 16 foram submetidas à maltagem por 24 e 48 horas e à extrusao obtendo-se flocos, os quais foram moídos e utilizados na fabricaçao de biscoitos. Flocos e biscoitos foram analisados sensorialmente. O efeito da maltagem e da extrusao foram também avaliados em relaçao às características químico-nutricionais. A composiçao em macro nutrientes nao apresentou alteraçoes acentuadas mostrando pequeno decréscimo nos teores de carboidratos totais e discreto aumento relativo nos teores de carboidratos totais e discreto aumento relativo nos teores de lipídeos no produto maltado. Dentre os fatores antinutricionais, os inibidores de tripsina apresentaram queda de 50 por cento após 24h de maltagem e desaparecimento após a extrusao. Os teores de ácido fítico nao sofreram variaçao expressiva, enquanto que os alfa-galactosídeos apresentaram decréscimo de até 80 por cento com os processos combinados de maltagem e extrusao.

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1. An experiment was conducted to determine the temperature for wet extrusion of full-fat soyabeans (FFS) needed to produce maximum chicken performance. 2. FFS were either unprocessed or extruded at 5 different temperatures (118 degrees, 120 degrees, 122 degrees, 126 degrees and 140 degrees C) in a wet extruder. Diets were prepared with the different FFS, and a diet prepared with soyabean meal (SBM) was included as a control. The 7 experimental diets were fed to individual groups of 40 chickens each, for a period of 35 d. Trypsin inhibitor activity (TIA), urease activity (UA), and protein solubility in potassium hydroxide (PS) were measured in all FFS and in the SBM. 3. Diets prepared with raw FFS and FFS extruded at 118 degrees and 120 degrees C resulted in significantly lower body weights and in pancreatic hypertrophy; maximum growth rate was obtained with FFS extruded at 122 degrees and 126 degrees C, while minimum pancreas weight was seen in chickens fed FFS extruded at 140 degrees C. 4. Although TIA, UA, and PS all decreased with increasing temperature of extrusion, TIA provided the best prediction of the feeding value of soyabeans for chickens.

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The chemical and toxicological composition of ten wild legumes collected in Yucatan, Mexico was determined. For each species the whole fruit, (seed and pod), were studied as well as the seed and pod separately. A higher protein content was found in the seeds of A. lebbeck and P. saman (37.07 and 37.60% respectively). In the seeds of L. longystilus, C. yucatanensis and P. keyense a high concentration of fat was found, especially in the first with 31.34%. A high quantity of fiber was found in the pods. In general, the samples were rich in lysine (especially seeds) and scant in sulfur amino acids and tryptophan. All the samples showed high concentration of potassium and calcium. Some of them exhibited significant concentrations of iron. The pods of P. saman and P. keyense showed a high content of lectins. In the seeds of C. yucatanensis and in the pod of P. keyense high concentrations of trypsin inhibitors were found 60 and 406.7 TUI/mg sample respectively. The presence of saponins, was detected in seven samples, of which the seed of P. keyense had the highest concentration. Alkaloids were found only in the whole fruit and pod of P. saman and cyanogenic glucosides were present in A. pennatula. In general terms, the whole legume showed better digestibility than the pods alone.

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Precooked flours obtained five Canavalia ensiformis varieties were prepared by dehydration in double drums. On a dry matter basis, significant differences (P < 0.05) among varieties were detected for crude protein content (25 to 30%), starch (36 to 40%) and dietary fiber (13 to 15%). Hemagglutinins were eliminated as result of the high temperature (146 degrees C/4 min) employed during the drying process. Similar results were not obtained for trypsin inhibitors and canavanine considering that small amounts of these compounds remained in the precooked flours prepared from canavalia seeds. A 10% decrease in available lysine was observed. Biological assays yielded Protein Efficiency Ratio (PER) values of 0, 8-1 and Neat Proteic Relation (NPR) values of 2-3-2.6. True digestibility of protein values were improved from 87 to 90%. All cultivars had similar starch digestive utilization coefficient (96%) and starch fraction (4%) resistant to enzymatic hydrolysis in the rat digestive tract.

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Precooked flours obtained five Canavalia ensiformis varieties were prepared by dehydration in double drums. On a dry matter basis, significant differences (P < 0.05) among varieties were detected for crude protein content (25 to 30 percent), starch (36 to 40 percent) and dietary fiber (13 to 15 percent). Hemagglutinins were eliminated as result of the high temperature (146 degrees C/4 min) employed during the drying process. Similar results were not obtained for trypsin inhibitors and canavanine considering that small amounts of these compounds remained in the precooked flours prepared from canavalia seeds. A 10 percent decrease in available lysine was observed. Biological assays yielded Protein Efficiency Ratio (PER) values of 0, 8-1 and Neat Proteic Relation (NPR) values of 2-3-2.6. True digestibility of protein values were improved from 87 to 90 percent. All cultivars had similar starch digestive utilization coefficient (96 percent) and starch fraction (4 percent) resistant to enzymatic hydrolysis in the rat digestive tract

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