Partial Characterization of Lectins Purified from the Surco and Vara (Furrow and Rod) Varieties of Black Phaseolus vulgaris.

As they manifest specifically and reversibly, lectins are proteins or glycoproteins with the characteristic of agglutinating erythrocytes. Given that grain legume lectins can represent 10% of protein content and can have various biological functions, they are extensively studied. The objective of this work was to purify and partially characterize the lectins of Phaseolus vulgaris black, var surco and vara (LBBS and LBBV). Both lectin types were purified by affinity chromatography on stroma matrix, which agglutinated human erythrocytes type A, B, and O, as well as rabbit, hamster, pig, and chicken erythrocytes. Native-PAGE was employed for molecular mass determination, yielding 109.36 and 112.68 kDa for BBS and BBV, respectively. Further analyses revealed that these lectins are tetrameric glycoproteins that require Ca+2, Mn+2 and Mg+2 ions for exhibiting their hemagglutinating function, which can be inhibited by fetuin. Moreover, optimal pH was established for both lectins (10.5 for LBBS and 7-9 for LBBV), while their activity was temperature-dependent and ceased above 70 °C. Finally, the observed differences in the biochemical characteristics and bioactive functions were ascribed to the different physiological characteristics of each seed, as well as the protein itself.

Bioremediation model for atrazine contaminated agricultural soils using phytoremediation (using Phaseolus vulgaris L.) and a locally adapted microbial consortium.

The objective of the present study was to examine a biological model under greenhouse conditions for the bioremediation of atrazine contaminated soils. The model consisted in a combination of phytoremediation (using Phaseolus vulgaris L.) and rhizopheric bio-augmentation using native Trichoderma sp., and Rhizobium sp. microorganisms that showed no inhibitory growth at 10,000 mg L-1 of herbicide concentration. 33.3 mg of atrazine 50 g-1 of soil of initial concentration was used and an initial inoculation of 1 × 109 UFC mL-1 of Rhizobium sp. and 1 × 105 conidia mL-1 of Trichoderma sp. were set. Four treatments were arranged: Bean + Trichoderma sp. (B+T); Bean + Rhizobium sp. (BR); Bean + Rhizobium sp. + Trichoderma sp. (B+R+T) and Bean (B). 25.51 mg of atrazine 50 g-1 of soil (76.63%) was removed by the B+T treatment in 40 days (a = 0.050, Tukey). This last indicate that the proposed biological model and methodology developed is useful for atrazine contaminated bioremediation agricultural soils, which can contribute to reduce the effects of agrochemical abuse.

Antioxidant Activity and Anticarcinogenic Effect of Extracts from Bouvardia ternifolia (Cav.) Schltdl.

According to the available ethnobotanical data, the Bouvardia ternifolia plant has long been used in Mexican traditional medicine to relieve the symptoms of inflammation. In the present study, the cytotoxic effect of extracts obtained from the flowers, leaves and stems of B. ternifolia using hexane, ethyl acetate (AcOEt) and methanol (MeOH) was evaluated by applying them to the SiHa and MDA-MB-231 cancer cell lines. An MTT reduction assay was carried out along with = biological activity assessments, and the content of total phenols, tannins, anthocyanins, betalains and saponins was quantified. According to the obtained results, nine extracts exhibited a cytotoxic effect against both the SiHa and MDA lines. The highest cytotoxicity was measured for leaves treated with the AcOEt (ID50 of 75 µg/mL was obtained for MDA and 58.75 µg/mL for SiHa) as well as inhibition on ABTS•+ against DPPH• radical, while MeOH treatment of stems and AcOEt of flowers yielded the most significant antioxidant capacity (90.29% and 90.11% ABTS•+ radical trapping). Moreover, the highest phenolic compound content was measured in the stems (134.971 ± 0.294 mg EAG/g), while tannins were more abundant in the leaves (257.646 mg eq cat/g) and saponins were most prevalent in the flowers (20 ± 0 HU/mg). Screening tests indicated the presence of flavonoids, steroids, terpenes and coumarins, as well as ursolic acid, in all the studied extracts. These results demonstrate the biological potential of B. ternifolia.

A semi-targeted NMR-based chemical profiling of retail samples of Mexican gordolobo.

Mexican gordolobo (MG) is one of the most important medicinal plants in Mexico and an important source of specialized metabolites with biological activities. The species employed in Mexican traditional medicine identified with this name are very different and difficult to identify according to existent criteria. In order to contribute to the identification of these species, a semi-targeted 1H NMR-based chemometric method was developed to characterize the chemical profiling of retail samples of MG. Thus, 17 retail samples of MG flowers were successively extracted with a gradient of water:chloroform (1:4, 1:2 and 0:1) in an ultrasonic bath for 15 min each. The organic phase of the three extracts was pooled, evaporated and analyzed by 1H NMR. The NMR data were subjected to multivariate statistical analysis, revealing that flavones gnaphaliin A (1), gnaphaliin B (2), araneol (3), 3,5,7-Tri-O-methylgalangin (4) and diterpenes sclareol (5) and kaur-16-en-18-oic acid (6) can be effectively used as chemotaxonomical markers for the correct identification of the Gnaphalium spp that compose MG. The aforementioned chemotaxonomic markers may be useful in establishing therapeutic criteria.

Lead, cadmium and cobalt (Pb, Cd, and Co) leaching of glass-clay containers by pH effect of food.

Recent studies have shown that handcrafted glass-clay containers are a health risk because they can be contaminated by heavy metals, which can be transferred to food, thus reaching the human body to potentially cause illness. Therefore, in the present work, we evaluate the leaching of lead, cadmium, and cobalt from glass-clay containers into two types of food: tomato sauce (salsa), and chickpea puree. The containers were obtained from four regions in the Mexican state of Hidalgo. Repetitive extractions from the containers were carried out to quantify the leaching of the heavy metals into the salsa, the chickpea puree, and acetic acid using the technique proposed by the USFDA. The results show that greater use of the containers leads to more leaching of heavy metals into both types of food and into the acetic acid, with the greatest metal extraction recorded for the Ixmiquilpan vessels. These results indicate that the metals present in the glass-clay containers leach into the food and that increased reuse increases the risk to the people who use them in food preparation.

Industrial application of catalytic systems for n-heptane isomerization.

The ideal gasoline must have a high pump octane number, in the 86 to 94 range, and a low environmental impact. Alkanes, as a family, have much lower photochemical reactivities than aromatics or olefins, but only the highly branched alkanes have adequate octane numbers. The purpose of this work is to examine the possibilities of extending the technological alternative of paraffin isomerization to heavier feedstocks (i.e., n-heptane) using non-conventional catalytic systems which have been previously proposed in the literature: a Pt/sulfated zirconia catalyst and a molybdenum sub-oxide catalyst. Under the experimental conditions at which these catalysts have been evaluated, the molybdenum sub-oxide catalyst maintains a good activity and selectivity to isomerization after 24 h, while the Pt/sulfated zirconia catalyst shows a higher dimethylpentanes/methylhexanes ratio, probably due to a lower operating temperature, but also a high formation of cracking products, and presents signs of deactivation after 8 h. Though much remains to be done, the performance of these catalysts indicates that there are good perspectives for their industrial application in the isomerization of n-heptane and heavier alkanes.

Purification, biochemical characterization, and bioactive properties of a lectin purified from the seeds of white tepary bean (phaseolus acutifolius variety latifolius).

The present work shows the characterization of Phaseolus acutifolius variety latifolius, on which little research has been published, and provides detailed information on the corresponding lectin. This protein was purified from a semi-domesticated line of white tepary beans from Sonora, Mexico, by precipitation of the aqueous extract with ammonium sulfate, followed by affinity chromatography on an immobilized fetuin matrix. MALDI TOF analysis of Phaseolus acutifolius agglutinin (PAA) showed that this lectin is composed of monomers with molecular weights ranging between 28 and 31 kDa. At high salt concentrations, PAA forms a dimer of 63 kDa, but at low salt concentrations, the subunits form a tetramer. Analysis of PAA on 2D-PAGE showed that there are mainly three types of subunits with isoelectric points of 4.2, 4.4, and 4.5. The partial sequence obtained by LC/MS/MS of tryptic fragments from the PAA subunits showed 90-100% identity with subunits from genus Phaseolus lectins in previous reports. The tepary bean lectin showed lower hemagglutination activity than Phaseolus vulgaris hemagglutinin (PHA-E) toward trypsinized human A and O type erythrocytes. The hemagglutination activity was inhibited by N-glycans from glycoproteins. Affinity chromatography with the immobilized PAA showed a high affinity to glycopeptides from thyroglobulin, which also has N-glycans with a high content of N-acetylglucosamine. PAA showed less mitogenic activity toward human lymphocytes than PHA-L and Con A. The cytotoxicity of PAA was determined by employing three clones of the 3T3 cell line, demonstrating variability among the clones as follows: T4 (DI50 51.5 µg/mL); J20 (DI50 275 µg/mL), and N5 (DI50 72.5 µg/mL).

Covalent Functionalization of Graphene Oxide with Fructose, Starch, and Micro-Cellulose by Sonochemistry.

In this work, we report the synthesis of graphene oxide (GO) nanohybrids with starch, fructose, and micro-cellulose molecules by sonication in an aqueous medium at 90 °C and a short reaction time (30 min). The final product was washed with solvents to extract the nanohybrids and separate them from the organic molecules not grafted onto the GO surface. Nanohybrids were chemically characterized by Fourier-transform infrared spectroscopy (FTIR), X-ray photoelectron spectroscopy (XPS), and Raman spectroscopy and analyzed by thermogravimetric analysis (TGA), scanning electron microscopy (SEM), and X-ray diffraction (XRD). These results indicate that the ultrasound energy promoted a chemical reaction between GO and the organic molecules in a short time (30 min). The chemical characterization of these nanohybrids confirms their covalent bond, obtaining a grafting percentage above 40% the weight in these nanohybrids. This hybridization creates nanometric and millimetric nanohybrid particles. In addition, the grafted organic molecules can be crystallized on GO films. Interference in the ultrasound waves of starch hybrids is due to the increase in viscosity, leading to a partial hybridization of GO with starch.

Incidence and behavior of Salmonella and Escherichia coli on whole and sliced zucchini squash (Cucurbitapepo) fruit.

The incidence of coliform bacteria (CB), thermotolerant coliforms (TC), Escherichia coli, and Salmonella was determined for zucchini squash fruit. In addition, the behavior of four serotypes of Salmonella and a cocktail of three E. coli strains on whole and sliced zucchini squash at 25+/-2 degrees C and 3 to 5 degrees C was tested. Squash fruit was collected in the markets of Pachuca city, Hidalgo State, Mexico. CB, TC, E. coli, and Salmonella were detected in 100, 70, 62, and 10% of the produce, respectively. The concentration ranged from 3.8 to 7.4 log CFU per sample for CB, and >3 to 1,100 most probable number per sample for TC and E. coli. On whole fruit stored at 25+/-2 degrees C or 3 to 5 degrees C, no growth was observed for any of the tested microorganisms or cocktails thereof. After 15 days at 25+/-2 degrees C, the tested Salmonella serotypes had decreased from an initial inoculum level of 7 log CFU to <1 log, and at 3 to 5 degrees C they decreased to approximately 2 log. Survival of E. coli was significantly greater than for the Salmonella strains at the same times and temperatures; after 15 days, at 25+/-2 degrees C E. coli cocktail strains had decreased to 3.4 log CFU per fruit and at 3 to 5 degrees C they decreased to 3.6 log CFU per fruit. Both the Salmonella serotypes and E. coli strains grew when inoculated onto sliced squash: after 24 h at 25+/-2 degrees C, both bacteria had grown to approximately 6.5 log CFU per slice. At 3 to 5 degrees C, the bacterial growth was inhibited. The squash may be an important factor contributing to the endemicity of Salmonella in Mexico.

Hibiscus Acid and Chromatographic Fractions from Hibiscus Sabdariffa Calyces: Antimicrobial Activity against Multidrug-Resistant Pathogenic Bacteria.

The anti-microbial properties of acetone extracts from Hibiscus sabdariffa calyces, fractions isolated by silica gel chromatography and hibiscus acid purified from some of these fractions and additionally identified by nuclear magnetic resonance spectroscopy, mid-infrared spectroscopy and X-ray diffraction, were studied against both multidrug-resistant Salmonella strains and pathogenic Escherichia coli bacteria. Gel diffusion was used to determine the anti-microbial effects. The mode of action of hibiscus acid was determined by crystal violet assay. Hibiscus acid and 17 of the 25 chromatographic fractions obtained, displayed an anti-microbial effect against all bacterial strains tested. Hibiscus acid showed a greater anti-microbial effect than the acetone extract against most of the bacteria strains, while chromatographic fractions IX-XIV exerted the greatest anti-microbial effect against all bacteria. The minimum inhibitory concentration of the acetone extract was 7 mg/mL, and the minimum bactericidal concentration was 10 mg/mL, while the corresponding values for hibiscus acid were 4-7 and 7 mg/mL, respectively. The results of the crystal violet assay indicate that hibiscus acid alters membrane permeability. Hibiscus acid is a potential alternative to control multidrug-resistant bacteria. Due to its ready availability and easy extraction from H. sabdariffa, hibiscus acid is potentially useful in the food industries.

Heat resistance of viable but non-culturable Escherichia coli cells determined by differential scanning calorimetry.

Several reports have suggested that the viable but non-culturable (VBNC) state is a resistant form of bacterial cells that allows them to remain in a dormant form in the environment. Nevertheless, studies on the resistance of VBNC bacterial cells to ecological factors are limited, mainly because techniques that allow this type of evaluation are lacking. Differential scanning calorimetry (DSC) has been used to study the thermal resistance of culturable bacteria but has never been used to study VBNC cells. In this work, the heat resistance of Escherichia coli cells in the VBNC state was studied using the DSC technique. The VBNC state was induced in E. coli ATCC 25922 by suspending bacterial cells in artificial sea water, followed by storage at 3 ± 2°C for 110 days. Periodically, the behaviour of E. coli cells was monitored by plate counts, direct viable counts and DSC. The entire bacterial population entered the VBNC state after 110 days of storage. The results obtained with DSC suggest that the VBNC state does not confer thermal resistance to E. coli cells in the temperature range analysed here.

Estudio químico de cinco plantas mexicanas de uso común en la medicina tradicional / Chemical study of five mexican plants used commonly in folk medicine

As a continuation of the phytochemical studies of Mexican medicinal plants, in this work we show the results of the investigation of five mexican plants that are commonly used in the folk medicine. From Gnaphalium sphacellatum Kunth was isolated obliquin, from G. inornatum DC, kaurenoic acid and beta-sitosterol, from G. purpurascens DC, gnaphaline and pinocembrin, from Spharalcea angustifolia (Cav.) G. Don, (–)-loliolide and sacharose and from Mimosa xanti A. Gray, beta-sitosterol, stigmasterol, (+)-pinitol, lupeol. The structures were elucidated by NMR and MS spectra.

Como una continuación de los estudios fitoquímicos de plantas medicinales de México, en este trabajo presentamos los resultados de la investigación de cinco plantas mexicanas comúnmente usadas en la medicina tradicional. De Gnaphalium sphacellatum Kunth se aisló obliquina, de G. inornatum DC, se aisló acido kaurenoico y beta-sitosterol, de G. purpurascens DC, gnaphalina y pinocembrina, de Sphaeralcea angustifolia, (Cav.) G. Don. (–)-loliolide y sacarosa y de Mimosa xanti A. Gray, beta-sitosterol, estigmasterol, (+)-pinitol, lupeol. Las estructuras fueron elucidadas por espectroscopía de RMN y MS.