Sustainable xanthophylls-containing poly(ε-caprolactone)s: synthesis, characterization, and use in green lubricants.

Three xanthophylls [(3R,3'R,6'R)-lutein (1), (3R,3'S)-zeaxanthin (2), and (3R,3'S)-astaxanthin (3)] were used for the first time as initiators in the ring-opening polymerization (ROP) of ε-caprolactone (CL) catalyzed by tin(ii) 2-ethylhexanoate [Sn(Oct)2] for the synthesis of novel sustainable xanthophyll-containing poly(ε-caprolactone)s (xanthophylls-PCL). The obtained polyesters were characterized by 1H and 13C NMR, FT-IR, DSC, SEC, and MALDI-TOF MS, and their use as additives in green lubricants was evaluated using a sliding friction test under boundary conditions. Xanthophylls-PCL were obtained with good conversions and with molecular weights determined by SEC to be between 2500 and 10 500 Da. The thermal properties of xanthophyll-polyesters showed a crystalline domain, detected by DSC. Lastly, the green lubricant activity of these polymers was evaluated and the results showed that xanthophylls-PCL could be employed as additives for biodegradable lubricant applications since they have better tribological behavior than current additives, which demonstrates their potential as future commercial materials with interesting eco-friendly properties for diverse applications.

Comparative Analysis of the Chemical Composition and Physicochemical Properties of the Mucilage Extracted from Fresh and Dehydrated Opuntia ficus indica Cladodes.

The development of sustainable extraction methods to obtain natural products constitutes a challenge for the food industry. The aim of this work was to compare yield, separation efficiency, chemical composition, and physicochemical properties of the mucilage extracted from fresh cladodes (FNM) and mucilage extracted from dehydrated cladodes (DNM) of O. ficus indica. Suspensions of fresh and dehydrated cladodes (4% w/w) were prepared for mucilage extraction by using a mechanical separation process. Subsequently, the separated mucilage was precipitated with ethyl alcohol (1:2 v/v) then, yield and separation efficiency were determined. The mucilage was characterized by measuring Z potential, viscosity, color, and texture attributes. Additionally, chemical proximate analysis, scanning electron microscopy, and thermogravimetric analysis (TGA) were conducted. No significant differences (p < 0.05) were detected in the yield and separation efficiencies between samples. Nevertheless, the dehydration process of cladodes prior to mucilage extraction increased protein, ashes, nitrogen free extract, and calcium content. The viscosity was higher in DNM than in FNM. The TGA revealed a different thermal behavior between samples. In addition, the DNM showed lower L (darkness/lightness), cohesiveness, adhesiveness, and springiness values than those of FNM. These results support that differences found between the chemical and physicochemical properties of DNM and those of FNM will determine the applications of the mucilage obtained from the O. ficus indica cladodes in the food, pharmaceutical, and cosmetic industries.

Glutamic Acid Increased Methotrexate Polyglutamation and Cytotoxicity in a CCRF-SB Acute Lymphoblastic Leukemia Cell Line.

Background and Objectives: Acute lymphoblastic leukemia (ALL) is the most common type of cancer in childhood. The majority of patients respond to treatment, but those with resistant phenotypes suffer relapse or death. The antifolate methotrexate (MTX) is the most commonly used drug against ALL due to its efficacy. Once inside leukemic cells, MTX is metabolized into methotrexate polyglutamates (MTX-PG) by action of the enzyme folylpolyglutamate synthetase (FPGS), leading to a longer action compared to that of MTX alone. Materials and Methods: In this work, we demonstrated that the combination treatment of methotrexate and 5 and 10 mM glutamic acid could enhance methotrexate cytotoxicity in CCRF-SB (B-ALL) cells. In addition, MTX plus 20 mM glutamic acid was able to improve the synthesis of MTX-PG5. Results: All treatments induced an increase in FPGS expression compared to that of the control group. Furthermore, we detected different cellular expression patterns of FPGS in the different treatments. Conclusion: Based on these findings, we demonstrated that levels of methotrexate polyglutamates (MTX-PGs) could be a key determinant of methotrexate-induced cytotoxicity in CCRF-SB acute lymphoblastic leukemia cells.

Study of the Interactions Occurring During the Encapsulation of Sesamol within Casein Micelles Reformed from Sodium Caseinate Solutions.

A casein micelle is a natural structure found in milk, based on the association between individual caseins and colloidal calcium phosphate, which can be used as vehicle for the encapsulation of hydrophobic compounds. In this project the capacity of micelles to encapsulate sesamol, a powerful antioxidant present in roasted sesame seeds, was evaluated. The micelles were reformed from sodium caseinate solutions at 2% or 5% (w/v) concentration, and then 1 or 2 mg/mL sesamol were added. A significant increase on the encapsulation efficiency was observed as caseinate concentration increased, going from 28% to 35% of sesamol encapsulated, while the encapsulation yield was greater in all cases for micelles from solutions with lower caseinate concentration. The average size of micelles ranged from 150 to 165 nm with an average zeta potential of -27.3 ± 1.86 mV. FTIR and fluorescence analysis confirm interactions within the casein chains and sesamol molecules with a bathochromic shift which suggests a predominant hydrophilic nature of such interactions. Differential scanning calorimetry thermograms showed that denaturation enthalpy tended to decrease as sesamol concentration increased, suggesting that sesamol molecules may be displacing the water molecules associated with the casein chains, reinforcing the idea of predominant hydrophilic interactions. Based on the results from encapsulation efficiency, it is estimated that about 7 g of casein micelles reformed from 2% (w/v) caseinate solutions with 2 mg/mL of added sesamol may provide the recommended daily dose and may be useful for the development of new functional food products. PRACTICAL APPLICATIONS: The development of a nanodelivery system for different bioactives will allow the enrichment of foods and drinks to develop new functional products that will satisfy consumers' demands. Additionally, the study of interactions between these molecules will allow us to understand how sesamol is being incorporated within the reformed micelles and how this process can even be improved.

Vasodilator Activity of Compounds Isolated from Plants Used in Mexican Traditional Medicine.

Arterial hypertension is one of the main risk factors in the development of cardiovascular diseases. Therefore, it is important to look for new drugs to treat hypertension. In this study, we carried out the screening of 19 compounds (triterpenes, diterpenes, sesquiterpenes, lignans, and flavonoids) isolated from 10 plants used in Mexican traditional medicine to determine whether they elicited vascular smooth muscle relaxation and, therefore, could represent novel anti-hypertension drug candidates. The vasorelaxant activity of these compounds was evaluated on the isolated rat aorta assay and the results obtained from this evaluation showed that three compounds induced a significant vasodilatory effect: meso-dihydroguaiaretic acid [half maximal effective concentration (EC50), 49.9 ± 11.2 µM; maximum effect (Emax), 99.8 ± 2.7%]; corosolic acid (EC50, 108.9 ± 6.7 µM; Emax, 96.4 ± 4.2%); and 5,8,4'-trihydroxy-3,7-dimethoxyflavone (EC50, 122.3 ± 7.6 µM; Emax, 99.5 ± 5.4%). Subsequently, involvement of the NO/cyclic guanosine monophosphate (cGMP) and H2S/ATP-sensitive potassium channel (KATP) pathways on the vasodilator activity of these compounds was assessed. The results derived from this analysis showed that the activation of both pathways contributes to the vasorelaxant effect of corosolic acid. On the other hand, the vasodilator effect of meso-dihydroguaiaretic acid and 5,8,4'-trihydroxy-3,7-dimethoxyflavone, partly involves stimulation of the NO/cGMP pathway. However, these compounds also showed an important endothelium-independent vasorelaxant effect, whose mechanism of action remains to be clarified. This study indicates that meso-dihydroguaiaretic acid, corosolic acid, and 5,8,4'-trihydroxy-3,7-dimethoxyflavone could be used as lead compounds for the synthesis of new derivatives with a higher potency to be developed as drugs for the prevention and treatment of cardiovascular diseases.

Nutraceutical Potential of New Alfalfa (Medicago sativa) Ingredients for Beverage Preparations.

Alfalfa (Medicago sativa) has been extensively used as animal feed, due to its fiber, protein, minerals, and vitamins, being also a useful source of phenolic compounds with potential therapeutic benefits. Nevertheless, its potential use as human ingredient is scarce. The aim of this work was to assess the nutritional composition, amino acid profile, and antioxidant capacity (AOC) of freeze-dried juice (FDJ) and fibrous residual material (RM), two new alfalfa-derived products (Adps) recently launched as ingredients for beverage preparations. Results demonstrated a high content of proteins (23-30 g/100 g FDJ and 13-17 g/100 g RM), crude fiber (29 g/100 g RM), and minerals (such as sodium, calcium, iron, and zinc). No significant difference was found in caloric content (4 kcal/g). Essential and nonessential amino acids were quantified in both Adps being leucine and lysine the most abundant. Total phenolic and total flavonoid contents (TPC and TFC, respectively) and their changes along the different harvesting periods of the year were also examined. FDJ presented the highest TPC in May (19 mg gallic acid equivalents/g dry weight [dw]), while in October TFC had the maximum value (4 mg catechin equivalents/g dw). Both products exhibited an interesting AOC by 1,1-diphenyl-2-picrylhydrazyl (DPPH) and ferric reducing antioxidant power assays. This study reports the nutraceutical potential of two new types of Adps.

Anti-inflammatory and toxicological evaluation of Moussonia deppeana (Schldl. & Cham) Hanst and Verbascoside as a main active metabolite.

ETHNOPHARMACOLOGICAL RELEVANCE: Moussonia deppeana, known as Tlachichinole, is a Mexican medicinal plant used for treatment of inflammatory diseases, influenza, diarrhea, gastrointestinal disorders and arthritis. AIM OF THE STUDY: In this paper the antioxidant and anti-inflammatory activities as well as the acute and sub-acute toxicological effects were evaluated for the ethanolic extract from aerial parts of M. deppeana, also its phytochemical analysis is described. MATERIALS AND METHODS: Phytochemical analysis and compound isolation were performed with thin layer chromatography. The chemical identification of the main compound was performed by (1)H NMR (COSY, NOESY, HSQC and HMBC) spectra. In vitro antioxidant capacity and total phenolic content for the ethanolic extract and its primary fractions was determined by DPPH and Folin-Ciocalteu reagent. Acute and subacute toxicity tests were evaluated on Balb/C mice. Finally acute anti-inflammatory evaluation was tested for a local (TPA) and systemic (carrageenan) murine model. RESULTS: The main compound isolated from the ethanolic extract of M. deppeana was Verbascoside, which was isolated from F3 and was identified by (1)H NMR and COSY data. Furthermore oleanolic and ursolic acids were isolated from primary fractions F1 and F2. Ethanolic extract showed IC50 = 6.71mg/mL for DPPH test and 664.12µg QE/mL for the total phenolic content. The LD50 value was >2g/kg by i.g. route in male and female mice. Sub-acute administration (28 days) of the ethanolic extract (1g/kg) did not cause lethality or alter any hematological and biochemical parameters, in addition, histological analysis of the major organs exhibited no structural changes. Anti-inflammatory activity of the ethanolic extract showed an ED50 = 1.5mg/ear and 450mg/kg for TPA and carrageenan test, respectively. Primary fractions generated moderate local and systemic anti-inflammatory activity. CONCLUSION: The ethanolic extract from the aerial parts of M. deppeana did not cause any lethality or adverse effect in either of the acute and sub-acute toxicity tests. This exhibited an important local and systemic anti-inflammatory activity and also moderate antioxidant capacity. Moreover, the primary fraction F2 was more active for the TPA model while the primary fraction F3 was most active in the carrageenan model in vivo. The main compound isolated from F3 was verbascoside; on the other hand also ursolic and oleanolic acids were isolated from F1 and F2.

Assessment of Beneficial and Possible Toxic Effects of Two New Alfalfa-Derived Shelf Products.

Aerial parts of Medicago sativa L. have been used as food and its consumption has been associated with health benefits, one among the most important being menopausal symptoms control. This work was aimed to explore possible pharmacological effects of two new alfalfa-derived products that have recently emerged as daily beverage preparations. In exploring their potential estrogenic effects, they produced no relevant alteration in the uterus. However, lowering glucose levels until normal values without causing further hypoglycemic effect were observed, when rats were treated with 1.5 g/kg/day samples. In vivo acute toxicity was not found when the alfalfa products were tested up to 3 g/kg rat weight. Furthermore, in vitro studies were conducted to assess their possible toxic effects. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and lactate dehydrogenase tests were carried out on the Caco-2 cell model to determine cell viability and membrane integrity. A concentration-dependent effect was observed, with a significant decrease in cell viability after exposure to concentrations of alfalfa product up to 100 mg/mL (after 3 h of incubation) and 50 mg/mL (after 24 h of treatment). Although in vitro level, the decrease in cell viability at these still low doses may underlie some toxicity, making necessary additional studies before any recommendation of a sustained consumption of these products by humans.

Systemic toxic effects induced by the aqueous extract of the fire coral Millepora complanata and partial purification of thermostable neurotoxins with lethal effects in mice.

Millepora complanata is a cnidarian widely distributed in the coral reefs of the Mexican Caribbean. This species is popularly known as "fire coral", since contact with it causes severe pain, skin eruptions and blisters. Intravenous administration of of M. complanata aqueous extract induces violent convulsions and death in mice within 1 min (LD50=4.62µgprotein/g of body weight). Doses less than the LD50 produced histopathological damage in kidneys and lungs. Such histopathological damage was completely eliminated after incubation of the extract in heat denaturing conditions. Unexpectedly, the denatured extract conserved its lethal effect. These findings demonstrated that the extract contained hemolytic and phospholipase activities that might be responsible for the histopathological damage, and additionally it contained other unidentified thermostable toxins with lethal effects in mice. Chromatographic analysis of the extract led to the isolation of a 61 kDa vasoconstrictor protein. Furthermore, several non-peptidic vasoconstrictor fractions were separated. Particularly interesting was the fraction MC1-IIA obtained as a result of three-step chromatography processes (ion exchange, gel filtration and reverse phase). Like the original crude extract, this fraction induced vasoconstriction and delayed hemolysis and lethal effects in mice. A subsequent chromatographic analysis of MC1-IIA showed that this fraction contained at least four non-peptidic compounds. MS and NMR spectroscopic data analyses indicated that these metabolites were poly-oxygenated alkylbenzenes. The present study constitutes the first report of the presence of non-peptidic lethal toxins in an organism of the class Hydrozoa, and evidences the great structural diversity of the toxins produced by the Millepora species.

Aortic relaxant activity of Crataegus gracilior Phipps and identification of some of its chemical constituents.

This study focused on the assessment of the vasorelaxant activity of the organic and aqueous extracts obtained from leaves and fruits of a Mexican hawthorn (Crataegus gracilior) on isolated rat aorta, and on the purification and identification of some of their secondary metabolites by the use of chromatographic and spectroscopic techniques. The results obtained showed that the methanol extract has a significantly more potent and effective vasorelaxant effect than the other tested extracts, with an EC50 = 8.69 ± 4.34 µg/mL and an Emax = 94.6% ± 11.30%, values that are close to that of acetylcholine, the positive control. From the same extract, two major triterpenes were isolated and identified as ursolic and corosolic acids by comparison of their experimental NMR spectroscopic data with those reported in the literature. Chlorogenic acid, rutin, quercetin, kaempferol and (+)-catechin were also identified using HPLC coupled with PDAD. All these compounds have already been proven to possess on their own antihypertensive effect and other benefits on cardiovascular diseases and they can support, at least in part, the traditional use of this plant species.

Anti-inflammatory and antioxidant activities of methanol extracts and alkaloid fractions of four Mexican medicinal plants of Solanaceae.

BACKGROUND: Methanol extracts and alkaloid fractions of different parts of four plant species belonging to Solanaceae family and used in Mexican traditional medicine were investigated for their total phenolic contents, anti-inflammatory and antioxidant properties. MATERIALS AND METHODS: The total phenolic compounds of each extract was determined according to the Folin-Ciocalteu method, while the in vitro radical scavenging activities of the extracts were assessed using the DPPH and ABTS radicals. The in vivo anti-inflammatory activity was determined using the TPA-induced mouse ear edema model. RESULTS: The methanol extracts contained the highest concentrations of phenolic compounds and also exhibited the best reducing power on the DPPH and ABTS radicals, in a concentration-dependent fashion. However, the anti-inflammatory activity did not follow the same trend, as some alkaloid fractions that showed low radical reducing power exhibited the strongest anti-inflammatory activity. CONCLUSION: The methanol extract obtained from the flowers of Nicotiana glauca presented the best overall performance with the largest amount of phenolic compounds (111 µg garlic acid equivalents/g of extract), the best antioxidant activity (94.80% inhibition of DPPH and 97.57% of ABTS) and the highest anti-inflammatory activity (81.93% inhibition of the inflammation).

Chemical constituents and antioxidant activity of extracts obtained from branch bark of Bursera simaruba / Constituyentes químicos y actividad antioxidante de extractos obtenidos de corteza de ramas de Bursera simaruba

The chemical constituents of the hexane and methanol extracts obtained from the branch bark of Bursera simaruba (Burseraceae) grown in Querétaro, Mexico, were investigated by GC-MS, HPLC coupled to DAD, and NMR techniques. Seventeen compounds, including terpenoids, flavonoids, phenolic acids, long-chain fatty acids (FA), methyl esters of FA and sucrose, were identified. In addition, an assessment of the antiradical activity of the methanol extract (ME) was also carried out using DPPH, ABTS, FRAP and DPV assays. The DPPH, ABTS and FRAP assays showed a low antioxidant capacity for the ME. This was in accordance with the relatively low quantities of phenols found in the extract. However, according to the differential pulse voltammetry assay (DVP), this extract exhibited an oxidation potential close to those of quercetin and (+)-catechin, two of the flavonoids with recognized good antioxidant power. This indicated that the ME does contain compounds with good antioxidant capacity and suggested that sometimes the most popular methods commonly used might be underestimating the true antioxidant capacities of plant samples and how the DPV is a valuable complementary tool to be taken into consideration when conducting these in vitro assays...

Los constituyentes químicos de los extractos hexánicos y metanólicos de la corteza de ramas de Bursera simaruba (Burseraceae) recolectada en Querétaro, México, fueron investigados mediante las Cromatografías de Gases acoplada a Espectrometría de masas (GC-MS) y de líquidos de alta resolución (HPLC) acoplada a un detector de arreglo de diodos (DAD) (HPLC-DAD) y mediante RMN. Diecisiete compuestos, incluyendo terpenos, flavonoides, ácidos fenólicos, ácidos grasos de cadena larga (AG), ésteres metílicos de AG y la sacarosa fueron identificados. De manera adicional, se determinó la actividad antioxidante del extracto metanólico utilizando los ensayos de DPPH, ABTS, FRAP y DPV (Voltametría de Pulso Diferencial). Los métodos de DPPH, ABTS y FRAP indicaron una baja capacidad antioxidante para este extracto. Esta observación estuvo de acuerdo con las bajas cantidades de fenólicos encontrados en este extracto. Sin embargo, mediante el método DPV, el extracto tuvo un potencial de oxidación cercano a los de la quercetina y la (+)-catequina, dos de los flavonoides con reconocida buna capacidad antioxidante. Estos resultados indican que el extracto sí contiene metabolitos secundarios con buena capacidad antioxidante y sugieren además que los métodos más comunes pueden subestimar la verdadera capacidad antioxidante de extractos de plantas y resalta la importancia del método DPV como complementario a tener en cuenta en estos tipos de estudios in vitro...

Intrapilosins I-VII, pentasaccharides from the seeds of Ipomoea intrapilosa.

Purification of a CHCl3-soluble extract from seeds of the Mexican medicinal arborescent morning glory, Ipomoea intrapilosa, by means of preparative-scale recycling HPLC, yielded seven new resin glycosides, intrapilosins I-VII (1-7). Their structures were established through the interpretation of their NMR spectroscopic and FABMS data. All pentasaccharides were found to be macrolactones of the known operculinic acid A with different fatty acids esterifying the same positions: C-2 on the second rhamnose unit and C-3 and C-4 on the third rhamnose moiety. The lactonization site of the aglycon could be placed at C-2 of the second saccharide. The fatty acid components of 1-7 were identified as (+)-(2S)-methylbutanoic, octanoic (caprylic), dodecanoic (lauric), and trans-cinnamic. The less common (-)-(2R)-methylbutanoic acid was also isolated as one of the saponification-liberated residues from intrapilosin IV (4). The presence of the (2R)- and (2S)-methylbutanoyl enantiomers bonded to the same oligosaccharide core in intrapilosins IV (4) and V (5) represents an example of diastereoisomerism due to a chiral esterifying moiety in the resin glycoside mixtures of a morning glory species.

Relationship among antimutagenic, antioxidant and enzymatic activities of methanolic extract from common beans (Phaseolus vulgaris L).

Common beans are rich in phenolic compounds, which can provide health benefits to the consumer. The objective of this work was to study the relationship among antimutagenicity, antioxidant and enzymatic activities of methanolic extract and trolox by principal components multivariate analysis. Antimutagenicity of phenolic compounds present in methanolic extract from the seed coat of common beans (P. vulgaris, Flor de Mayo Bajío cultivar) and trolox against AFB1 mutagenicity were evaluated in the Salmonella typhimurium microsuspension assay. Antioxidant capacity of methanolic extract and trolox were evaluated using beta-carotene and 1,1-diphenyl-2-picryhydrazyl (DPPH) in vitro model assays. Cythrome P450 activity was measured by fluorometric assay. For phenolic extract, trolox and phenolic extract+trolox, the inhibition on AFB1 mutagenicity in tester strain TA100 was 47, 59 and 69%, respectively. While in TA98 was 39, 48 and 68%. The inhibition of phenolic compounds, trolox and phenolic compounds+trolox on cytochrome P450 (CYP450) activity was 48, 59 and 88%, respectively. Correlation analysis showed that phenolic extract and trolox have high antimutagenic and antioxidant activity and also inhibited enzymatic activity. The results suggest that the primary mechanism of action of phenolic compounds in beans against AFB1 mutagenicity may be extra-cellular in the microsuspension assay.

Biodynamic constituents in the Mexican morning glories: purgative remedies transcending boundaries.

Jalap, a pre-Hispanic herbal remedy still considered a useful laxative, is an ingredient in some over-the-counter products sold by herbalists in contemporary Mexico. The purgative crude drugs are prepared from the roots of several morning glories species which all have been identified as members of the Ipomoea genus (Convolvulaceae). Their incorporation as therapeutical agents into Europe is an outstanding example of the assimilation of botanical drugs from the Americas as substitutes for traditional Old World remedies. Phytochemical investigations on the resin glycosides, main constituents of these drugs, were initiated during the second half of the XIX century; however, the structures of their active ingredients had remained poorly known, and still are for some members of these purgative root species. Modern analytical techniques with higher resolution capabilities (HPLC) for the isolation of the active principles of these crude drugs used in conjunction with powerful spectroscopic methods (high field NMR) have facilitated the investigation of these relevant, to the herbal product market, convolvulaceous species during the last decade. The advantages and limitations of theses techniques will be discussed. This review will also describe the ethnobotanical information associated with the Mexican morning glory species and how the traditional usages of these plants have played an important role in the selection of these materials for chemical studies. Little is as yet known about either the mechanism of purge action caused by the resin glycosides or the ecological significance of these same compounds for the producing plants. Over the five centuries of Mexican herbal medicine, one hundred years of phytochemistry has only partially elucidated the active ingredients of the jalap roots but has exemplified how to further contemporary drug discoveries through the investigation of those plants traditionally held to be economically and medicinally important in developing countries.

In vitro inhibition of 1,3-beta-glucan synthase by glycolipids from convolvulaceous species.

Sixteen convolvulaceous glycolipids selected from the tricolorin (1 - 7) and orizabin (8 - 16) series, proved to be strong in vitro inhibitors of the enzyme that catalyzes the synthesis of 1,3-beta-D-glucan, a major polymer of fungal cell-walls. Results provide an insight into function of the specific structures of these complex macrocyclic lactones as inhibitors of the 1,3-beta-D-glucan synthase and open the possibility of using these compounds as starting points for the development of antifungal agents that act by inhibiting fungal cell-wall synthesis.