Unraveling the effects of drying techniques on chaya leaves: Metabolomics analysis of nonvolatile and volatile metabolites, umami taste, and antioxidant capacity.

Chaya (Cnidoscolus chayamansa) leaves are known for their strong umami taste and widespread use as a dried seasoning. This study aimed to assess the impact of different drying methods [freeze drying (FD), vacuum drying, oven drying at 50 °C and 120 °C (OD120) and pan roasting (PR)] on the metabolome using mass spectrometry, umami intensity, and antioxidant properties of chaya leaves. The predominant volatile compound among all samples, 3-methylbutanal, exhibited the highest relative odor activity value (rOAV), imparting a malt-like odor, while hexanal (green grass-like odor) and 2-methylbutanal (coffee-like odor) are the second highest rOAV in the FD and PR samples, respectively. OD120 and PR samples possessed the highest levels of umami-tasting amino acids and 5'-ribonucleotides as well as the most intense umami taste, whereas FD samples exhibited the highest antioxidant capacity. These findings enhance our understanding of the aroma characteristics, umami taste, and antioxidant potential of processed chaya leaves.

Glycemic Index and Glycemic Load of Two Dishes Cooked with Alache (Anoda cristata) and Chaya (Cnidoscolus aconitifolius) Plants from the Traditional Mexican Diet.

Correct nutrition is important for keeping good health; to attain that, the diet has to include vegetables such as quelites. The objective of this study was to determine the glycemic index (GI) and glycemic load (GL) of rice and a tamal prepared with and without two species of quelites: "alache" (Anoda cristata) and "chaya" (Cnidoscolus aconitifolius). The GI was measured in 10 healthy subjects, 7 women and 3 men, with the following mean metrics: age, 23 years old; body weight, 61.3 kg; height, 1.65 m; body mass index, 22.7 kg/m2; and basal glycemia, 77.4 mg/dL. Capillary blood samples were collected within 2 h after the meal. White rice (rice with no quelites) had a GI of 75.35 ± 15.6 and a GL of 36.17 ± 7.8; rice with alache had a GI of 33.74 ± 5.85 and a GL 33.74 ± 1.85. White tamal had a GI of 57.33 ± 10.23 and a GC of 26.65 ± 5.12; tamal with chaya had a GI of 46.73 ± 22.1 and a GL of 23.36 ± 11. The GI and GL values recorded for the combination of quelites with rice and tamal confirmed that quelites could be a good alternative for healthy diets.

Effects of dried chaya leaf meal inclusion in the diet on growth performance and blood profiles in Thai native chicken (Pradu Hangdum).

Objective: The objective of the present study was to evaluate the effects of using dried chaya leaf meal (DCLM) as a protein feed in a diet on growth performance, blood profiles, and carcass quality in a native Thai chicken. Materials and Methods: Eighty 14-day-old Pradu Hangdum chicks were divided into four groups with four replicates each: control (without DCLM), 10%, 20%, and 30% DCLM inclusion in mash feed. Growth performance was recorded weekly until 98 days of age. Blood profile, carcass quality, and visceral organ weight were measured at 98 days of age. Results: The 10%-30% DCLM inclusion in the diet did not affect feed intake or feed efficiency; however, the body weight gain of chicks decreased linearly with the increase in DCLM inclusion. The heterophils, eosinophils, and monocytes were linearly increased with the increasing DCLM levels in the groups. The serum blood chemistry did not differ among the groups, while the AST in 10% and 20% DCLM was lower than in the control. Increasing the level of DCLM in the chicken diet did not affect carcass quality. Conclusion: The DCLM can be used as a feed ingredient in Thai native chicken feed up to 20%.

Chaya (Cnidoscolus aconitifolius (Mill.) I.M. Johnst) leaf extracts regulate mitochondrial bioenergetics and fatty acid oxidation in C2C12 myotubes and primary hepatocytes.

ETHNOPHARMACOLOGICAL RELEVANCE: Chaya (Cnidoscolus aconitifolius (Mill.) I.M. Johnst) is an important component of the regular diet and traditional medicine of indigenous communities in Mexico. Customarily, Chaya is consumed as a beverage made of macerated leaf, cooked, or prepared in teas or infusions to empirically treat obesity, diabetes, gastrointestinal disorders, and kidney stones. The beneficial effects of Chaya can be attributed to the presence of protein, dietary fiber, vitamins, and especially polyphenols, which regulate mitochondrial function. Therefore, polyphenols present in Chaya extracts could be used to develop novel strategies to prevent and treat metabolic alterations related to mitochondrial dysfunction in the muscle and liver of subjects with obesity, type 2 diabetes, and cardiovascular diseases. However, limited information is available concerning the effect of Chaya extracts on mitochondrial activity in those tissues. AIM OF THE STUDY: The aim of this study was to evaluate the antioxidant capacity of an aqueous extract (AE) or mixed (methanol/acetone/water) extract (ME) of Chaya leaf and their effect on C2C12 myotubes and primary hepatocyte mitochondrial bioenergetics and fatty acid oxidation (FAO). MATERIALS AND METHODS: Total polyphenol content and antioxidant activity were determined using the Folin-Ciocalteu method and the oxygen radical absorbance capacity assay, respectively. The effect of AE and ME from Chaya leaf on mitochondrial activity and FAO of C2C12 myotubes and primary hepatocytes was evaluated using an extracellular flux analyzer. RESULTS: The AE and ME from Chaya leaf exhibited antioxidant activity and a polyphenol content similar to nopal, another plant used in Mexican traditional medicine. AE significantly (p < 0.05) decreased the maximal respiration and spare respiratory capacity (SRC) of C2C12 cells, whereas ME had little effect on C2C12 mitochondrial function. Conversely, ME significantly (p < 0.05) decreased SRC in primary hepatocytes, whereas AE increased maximal respiration and SRC at low doses (5 and 10 µM). Moreover, low doses of Chaya AE significantly (p < 0.05) increased AMPK phosphorylation, acyl-coenzyme A oxidase protein abundance, and palmitate oxidation in primary hepatocytes. CONCLUSION: The AE of Chaya leaf increases mitochondrial function and FAO of primary hepatocytes, indicating its potential to treat hepatic mitochondrial dysfunction underlying metabolic diseases.

Metabolite profiling and identification of novel umami compounds in the chaya leaves of two species using multiplatform metabolomics.

Chaya (Cnidoscolus chayamansa and C. aconitifolius) is a fast-growing medicinal plant, and its leaves exhibit a strong umami taste. Here metabolite variation and umami-related compounds in the leaves of two chaya species were determined using a multiplatform untargeted-metabolomics approach, electronic tongue, and in silico screening. Metabolite profiles varied between the leaves of the two species and among leaf maturation stages. Young leaves exhibited the highest umami taste intensity, followed by mature and old leaves. Partial least square regression and computational molecular docking analyses revealed five potent umami substances (quinic acid, trigonelline, alanyl-tyrosine, leucyl-glycyl-proline, and leucyl-aspartyl-glutamine) and three known umami compounds (l-glutamic acid, pyroglutamic acid, and 5'-adenosine monophosphate). The five substances were validated as novel umami compounds using electronic tongue assay; leucyl-glycyl-proline exhibited synergism with monosodium glutamate, thereby enhancing the umami taste. Thus, substances contributing to the taste of chaya leaves were successfully identified.

Protective Effects of Chaya against Mitochondrial and Synaptic Toxicities in the Type 2 Diabetes Mouse Model TallyHO.

The purpose of our study is to determine the protective effects of the chaya leaf against mitochondrial abnormalities and synaptic damage in the Type 2 diabetes (T2D) mouse model, TallyHO (TH). The TH mouse is a naturally occurring polygenic mouse model of diabetes that mimics many characteristics of human Type 2 diabetes. Only male TH mice develop hyperglycemia and moderate obesity. Female mice display moderate obesity but do not manifest overt diabetes. In this study, we evaluated three groups of mice over a period of 11 weeks: (1) the experimental group of TH diabetic mice fed with chaya chow; (2) a diabetic control group of TH diabetic mice fed with regular chow; and (3) a non-diabetic control group of SWR/J mice fed with regular chow. Body mass and fasting blood glucose were assessed weekly. Brain and other peripheral tissues were collected. Using qRT-PCR and immunoblotting analyses, we measured the mRNA abundance and protein levels of mitochondrial biogenesis, mitochondrial dynamics, autophagy/mitophagy, and synaptic genes. Using immunofluorescence analysis, we measured the regional immunoreactivities of mitochondrial and synaptic proteins. Using biochemical methods, we assessed mitochondrial function. We found increased body mass and fasting glucose levels in the TH diabetic mice relative to the non-diabetic control SWRJ mice. In chaya chow-fed TH diabetic mice, we found significantly reduced body mass and fasting glucose levels. Mitochondrial fission genes were increased and fusion, biogenesis, autophagy/mitophagy, and synaptic genes were reduced in the TH mice; however, in the chaya chow-fed TH diabetic mice, mitochondrial fission genes were reduced and fusion, biogenesis, autophagy/mitophagy, and synaptic genes were increased. Mitochondrial function was defective in the diabetic TH mice; however, it was rescued in the chaya chow-fed TH mice. These observations strongly suggest that chaya chow reduces the diabetic properties, mitochondrial abnormalities, and synaptic pathology in diabetic, TH male mice. Our data strongly indicates that chaya can be used as natural supplemental diet for prediabetic and diabetic subjects and individuals with metabolic disorders.

The complete chloroplast genome sequence of chaya (Cnidoscolus aconitifolius) and phylogenetic analysis.

Cnidoscolus aconitifolius is a leafy green heathy vegetable and medicinal plant belongs to the family Euphorbiaceae. In the present study, we sequenced the complete chloroplast genome of C. aconitifolius, which is 158,658 bp in length and consisted of two copies of inverted repeat (IR) of 26,982 bp separated by a large single copy (LSC) of 87,022 bp and a small single copy (SSC) of 17,672 bp. The GC content of C. aconitifolius was 36.3%. A total of 130 genes were predicted, including 86 protein-coding genes, 36 tRNAs and 8 rRNAs. The plastid phylogenomic analysis support C. aconitifolius is closely related to Manihot esculenta.

Chaya Leaf Decreased Triglycerides and Improved Oxidative Stress in Subjects With Dyslipidemia.

Chaya is an edible leaf popular in Mexico and Central America because of its high nutritional value. Studies in animal models have demonstrated the beneficial effects of Chaya, which include reduction of circulating lipids and increase in antioxidant activity. However, its hypolipidemic and antioxidant effects have not been demonstrated in humans. Thus, the aim of the present study was to evaluate the effect of Chaya on the lipid profile, lipid peroxidation, inflammation, and peripheral blood mononuclear cell gene expression in a population with dyslipidemia. We performed a single-arm trial in 30 participants with dyslipidemia who consumed 500 mL of Chaya beverage per day over a 6-week period. Interestingly, we observed a significant decrease in serum triglyceride concentration (P < 0.05) and an increase in plasma antioxidant activity and polyphenol concentration (P < 0.005) after 6 weeks of Chaya consumption. This was accompanied by a reduction in the oxidative stress marker MDA (P < 0.0001) and by an increase in the antioxidant enzyme CAT expression in peripheral blood mononuclear cells (P < 0.001). Altogether, our results demonstrate that consumption of Chaya has hypotriglyceridemic and antioxidant effects in subjects with dyslipidemia.

Cnidoscolus aconitifolius leaf pellet can manipulate rumen fermentation characteristics and nutrient degradability.

OBJECTIVE: Chaya (Cnidoscolus aconitifolius) leaf has been found to be an important source of protein, vitamins, minerals, as well as phytonutrients. The present study aimed to evaluate the effect of Chaya leaf pellet (CHYP) with various level of crude protein (CP) in the concentrate on rumen fermentation characteristics and nutrient degradability in in vitro gas production technique. METHODS: In an in vitro rumen fermentation study the dietary treatments were arranged according to a 3×5 factorial arrangement in a completely randomized design, consisting of Factor A: three levels of CP of concentrate mixtures (14%, 16%, and 18% CP, respectively) and Factor B: five levels of CHYP supplementation (at 0%, 2%, 4%, 6%, and 8% of dry matter substrates). RESULTS: The gas production kinetics, fraction (a) and fraction (b) were lower (p<0.05) with an increasing CHYP addition. Additionally, the fraction (a+b) was found to yield a significant interaction (p<0.05) while the fraction (c) was not impacted by CHYP addition. However, in vitro DM degradability was enhanced and interactive (p<0.05), using 16% CP of concentrate with 6% and 8% CHYP, when compared with 18% CP in the non-addition. Additionally, the treatment with higher CP of the concentrate was higher in NH3-N concentration (p<0.001) and by CHYP supplementation group (p<0.05). Nevertheless, protozoal counts in the rumen were remarkably decreased (p<0.05) with increasing level of CHYP supplementation. Furthermore, rumen C2 concentration was lower (p<0.05) in the treatments with CHYP supplementation, while C3 was significantly increased and interactive (p<0.05) between levels of CP and CHYP supplementation especially at 8% CHYP supplementation. CONCLUSION: Based on this study, the results revealed CHYP as a promising feed supplement to enhance rumen fermentation and to mitigate methane production. However, in vivo feeding experiments should be subsequently conducted to elucidate the effect of CHYP supplementation on rumen fermentation, as well as ruminant production efficiency.

Herbivory and anti-herbivore defences in wild and cultivated Cnidoscolus aconitifolius: disentangling domestication and environmental effects.

Phenotypic changes in plants during domestication may disrupt plant-herbivore interactions. Because wild and cultivated plants have different habitats and some anti-herbivore defences exhibit some plasticity, their defences may be also influenced by the environment. Our goal was to assess the effects of domestication and the environment on herbivory and some anti-herbivore defences in chaya (Cnidoscolus aconitifolius) in its centre of domestication. Herbivores, herbivory, and direct and indirect anti-herbivore defences were assessed in wild and cultivated plants. The same variables were measured in the field and in a common garden to assess environmental effects. Our results show that domestication increased herbivory and herbivore abundance, but reduced direct and some indirect defences (ants). The environment also affected the herbivore guild (herbivore abundance and richness) and some direct and indirect defences (trichome number and ants). There was also an interaction effect of domestication and the environment on the number of trichomes. We conclude that domestication and the environment influence herbivory and anti-herbivore defences in an additive and interactive manner in chaya.

Reproductive isolation between wild and domesticated chaya (Cnidoscolus aconitifolius) in sympatry.

Reproductive isolation is a necessary condition for plant domestication in their domestication centre where crops co-occur with their wild progenitors. However, the identification of reproductive barriers and their relative contribution to reproductive isolation have been overlooked in plants under domestication. We assessed pre- and post-pollination reproductive barriers and their relative contribution to reproductive isolation between wild and domesticated chaya (Cnidoscolus aconitifolius) in its domestication centre. We found that wild and domesticated chaya both exhibit a high degree of reproductive isolation. However, the reproductive isolation barriers exhibited some asymmetry: while pre-pollination barriers (differential pollen production and pollinator specificity) were only detected in wild plants, post-pollination barriers (pollen-pistil incompatibility and/or failure to set fruit) were observed in both wild and domesticated plants. We conclude that complete reproductive isolation has evolved in sympatry in co-occurring domesticated and wild chaya.

Effect on hyperglycemia and pancreas cells of chaya aqueous extracts from two different regions in streptozotocin-induced diabetes rats

Cnidoscolus chayamansa is a native plant of the Mayan region, which is also cultivated in other places like northern Mexico, Tunisia and India. Many properties are attributed to Mayan Chaya, such as aid in the control of glycemia in diabetics. Thus this study aimed to evaluate the hypoglycemic effects of chaya aqueous extracts in a model of streptozotisin-induced diabetic Wistar rats. Chaya aqueous extracts were collected from plants cultivated in Quinta Roo (Mayan region) and Durango (northern Mexico), and in this study we compare their effect with metformin (as a control). Additionally, we compared the extracts mass profiles from both regions by high-resolution liquid chromatography coupled to a triple quadrupole tandem mass detector (HPLC-MS/MS QQQ). Finally, a study of the pancreatic tissue was carried out to evaluate the effects of the extracts on the Langerhans islets. Both extracts showed a good hypoglycemic effect after two weeks of treatment, and the Langerhans islets showed a partial recovery due to the effect of the treatment. Although the plants were cultivated at a distance of 2,350 km and under different weather, the compounds found in both did not show significant differences.

Fitoquímica preliminar, actividad antioxidante e hipoglucemiante de extractos de hojas de Cnidoscolus aconitifolius (Mill.) I. M. Johnst (chaya) / Phytochemical, antioxidant and hypoglucemic activity of extracts from leaves of Cnidoscolus aconitifolius (Mill.) I. M. Johnst (chaya)

Objetivo: evaluar, preliminarmente, la composición química de diferentes extractos obtenidos a partir de las hojas de Cnidoscolus aconitifolius (Mill.) I. M. Johnst que crece en Ecuador, así como la actividad antioxidante e hipoglucemiante del extracto acuoso. Métodos: el estudio fitoquímico de la droga cruda fue realizado mediante Cromatografía gaseosa acoplada a Espectrometría de Masa y Resonancia Magnética Nuclear Protónica a diferentes extractos obtenidos por maceración en hexano, acetona y metanol. La evaluación antioxidante del extracto acuoso incluyó determinaciones del potencial de reducción total, de glutatión reducido y organoperóxidos totales. La actividad hipoglucemiante del extracto acuoso liofilizado se estudió, en ratas, utilizando alloxano como agente inductor de la hiperglucemia. Resultados: se identificaron los principales metabolitos presentes en el material vegetal, resaltando los ácidos grasos (ácido palmítico, ácido linoleico, ácido ?-linoleico), triterpenos (3 ceto-?-amirina, ?-amirina, acetato de ?-amirina y acetato de lupeol) y azúcares (glicerol, xilitol, arabitol, glucitol). En cuanto a las potencialidades de empleo de la planta: la actividad antioxidante para los grupos -SH no proteicos del extracto acuoso de droga cruda a concentraciones de 0,12 mg/mL mostró valores significativos en relación con los intervalos de referencia en plasma humano. Sin embargo, el extracto acuoso liofilizado obtenido a partir de la droga cruda, a la dosis de 500 mg/kg de peso corporal de la rata, no demostró poseer efecto hipoglucemiante(AU) Conclusiones: de manera preliminar se identificaron los principales metabolitos presentes en la especie que crece en Ecuador y las posibilidades de empleo del extracto acuoso como agente antioxidante, no así como hipoglucemiante(AU)

Objective: to evaluate, preliminary, chemical composition of different extracts obtained from leaves of Cnidoscolus aconitifolius (Mill.) I. M. Johns, and antioxidant and hypoglycemic activity of water extract. Methods: phytochemical study to different extracts obtained by maceration in hexane, acetone and methanol was conducted by Gas chromatography-mass spectrometry and Proton Nuclear Magnetic Resonance. Antioxidant evaluation of water extract included total reduction potential, reduced glutathione and organoperoxides determinations. The hypoglycemic activity of lyophilized water extract was studied in rats using alloxano as hyperglycemia-inducing agent. Results: the main metabolites present in the plant were identified, fatty acids (palmitic acid, linoleic acid, ?-linolenic acid), triterpenes (3 keto ?-amyrin, ?-amyrin, ?-amyrin acetate and ?-lupeol acetate) and sugars (glycerol, xylitol, arabitol, glucitol). For potential use of the plant: antioxidant activity for non-protein -SH groups of the water extract of raw drug at concentrations of 0,12 mg/mL showed significant values compared to the reference ranges in human plasma. However, the lyophilized water extract obtaines from de raw drug (dose of 500 mg/kg body weight of rat) was shown to possess hypoglycemic effect. Conclusions: were identified, preliminary, the mayor metabolites present in the species growing in Ecuador and possible use of water extract as antioxidant agent and not like hipoglucemian(AU)