Extraction of bioactive compounds from pecan nutshell: An added-value and low-cost alternative for an industrial waste.

The pecan nutshell [Carya illinoinensis (Wangenh) C. Koch] (PNS) is a source of bioactives with important beneficial properties for the human health. PNS represents between 40-50 % of total mass of the nut, resulting as waste without any added value for the food industry. Even though a variety of methods were already developed for bioactive extraction from this waste, unconventional methodologies, or those which apart from green chemistry principles, were discarded considering the cost of production, the sustainable development goals of United Nations and the feasibility of real inclusion of the technology in the food chain. Then, to add-value to this waste, a low-cost, green and easy-scalable extraction methodology was developed based on the determination of seven relevant factors by means of a factorial design and a Response Surface Methodology, allowing the extraction of bioactives with antioxidant capacity. The pecan nutshell extract had a high concentration of phenolic compounds (166 mg gallic acid equivalents-GAE/g dry weight-dw), flavonoids (90 mg catechin equivalent-CE/g dw) and condensed tannins (189 mg CE/g dw) -related also to the polymeric color (74.6 %)-, with high antioxidant capacities of ABTS+. radical inhibition (3665 µmol Trolox Equivalent-TE/g dw) and of iron reduction (1305 µmol TE/g dw). Several compounds associated with these determinations were identified by HPLC-ESI-MS/MS, such as [Epi]catechin-[Epi]catechin-[Epi]gallocatechin, myricetin, dihydroquercetins, dimers A and B of protoanthocyanidins, ellagitannins and ellagic acid derivatives. Hence, through the methodology developed here, we obtained a phenolic rich extract with possible benefits for human health, and of high industrial scalability for this co-product transformation.

Proteomic and Transcriptomic Analyses Provide New Insights into the Mechanism Underlying Lipid Deterioration in Pecan Kernels during Storage.

Pecan nuts are rich in lipids that tend to deteriorate during storage. Tandem mass-tag-based quantitative proteomics and transcriptomics were used to investigate the changes in the protein and gene profiles of stored pecan kernels for the first time. Our previous lipidomic data were jointly analyzed to elucidate the coordinated changes in lipid molecules and related proteins/genes. The mechanism underlying lipid deterioration in pecan kernels during storage was revealed by multiomics analyses. Lipid metabolism-related pathways were activated during pecan storage. Phospholipases, triacylglycerol lipases, lipoxygenases, and oil body-related proteins/genes were highly expressed during storage, revealing their involvement in lipid deterioration. These data provide rich information and will be valuable for future genetic or chemical research to alleviate lipid deterioration in pecans.

Quantitative determination of specialised metabolites in different parts of Juglans regia Linn. and Carya illinoinensis (Wangenh.) K. Koch by using UHPLC-DAD-QTOF-MS/MS.

INTRODUCTION: Juglans regia Linn. and Carya illinoinensis (Wangenh.) K. Koch are nut-producing plant species of the Juglandaceae family. Bioactive compounds like naphthoquinones, tetralones, and diarylheptanoids are dominant in these species. OBJECTIVE: The study aimed to develop and validate a fast and sensitive analytical method by ultrahigh-performance liquid chromatography diode array detection mass spectrometry (UHPLC-DAD-MS) for quantification and identification of bioactive compounds in fruit pericarps and leaves of J. regia and C. illinoinensis collected from two different states of north India. METHODOLOGY: The dried pericarps of J. regia and C. illinoinensis (500 mg) were extracted with ethyl acetate-methanol (50:50 v/v, 20 mL, 50°C, 30 min) by ultrasonication and analysed by ultrahigh-performance liquid chromatography diode array detection quadrupole time-of-flight tandem mass spectrometry (UHPLC-DAD-QTOF-MS/MS) for qualitative and quantitative examination of phytoconstituents. The method was validated according to International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human use (ICH) guidelines for linearity, precision, accuracy, limit of detection (LOD), and limit of quantification (LOQ). RESULTS: Here, we report the quantification of dihydrophaseic acid (1), 4,5-dihydroxytetralone (2), 4,8-dihydroxytetralone (3), 5,8-dihydroxy-3-methoxytetralone (4), and juglanin A (5) in the pericarps and leaves of J. regia and C. illinoinensis. Furthermore, using the hyphenated analytical method, a total of 62 compounds were tentatively characterised in different samples. The principal component analysis (PCA) showed diversity between the analysed sample's composition. Also, the study evaluated the variation of bioactive compounds among different parts of J. regia and C. illinoinensis collected from different regions of northern India by UHPLC-DAD-QTOF-MS/MS. CONCLUSION: The developed method is simple, rapid, and selective for the identification and characterisation of bioactive molecules.

Targeted metabolomics reveals key phenolic changes in pecan nut quality deterioration under different storage conditions.

Pecan nuts are highly enriched in phenolic compounds, which contribute to the health benefits of pecans. Phenolic compounds represent the main oxidation reaction substrates, thus leading to quality deterioration, namely pellicle browning or a decrease in beneficial effects during pecan storage. Hence, four different storage conditions were performed for 180 d to simulate real production situations. Targeted metabolomics was chosen to identify the specific phenolic compounds involved in quality deterioration under different storage conditions in 0, 90, and 180 d samples. A total of 118 phenolic compounds were detected, nine of which were identified for the first time in pecan. The total phenolic content (TPC) and antioxidant capacities initially demonstrated high scores, after which they tended to decrease during the storage process. The significantly modified phenolic compounds during storage were selected as the metabolite markers of pecan quality deterioration, including catechin, procyanidin (PA) trimer, PA tetramer, trigalloyl hexahydroxydiphenoyl (HHDP) glucose, and tetragalloyl hexoside. Fresh pecan kernels resulted in more pronounced changes in hydrolysable tannins (HTs), whereas dry kernels resulted in the most accentuated changes in condensed tannins (CTs). To the best of our knowledge, this is the first attempt to study individual phenolic changes during storage of pecan in such massive amounts. The results can offer a valuable theoretical basis for future control of pecan quality deterioration through phenolics during storage.

Changes in chemical characteristics and modeling sensory parameters of stored pecan nutmeats.

Pecan is a major specialty crop produced in the United States. Sensory evaluation and chemical analyses of pecan nutmeats are integral components of shelf life and have been employed to investigate changes during storage, but there remains a lack of knowledge regarding storage stability. Specifically, the association between shelf life and chemical characteristics has not been investigated. We aimed to investigate the chemical changes in pecan nuts during a range of storage treatments (temperature, relative humidity, packaging material, and modified atmosphere). The results of the chemical analyses were used to build a volatile compound-based sensory prediction model. The work has utility as a rapid method to measure lipid oxidation in pecan, which is of value to the pecan industry. The research also determined a possible association between pecan nut volatile compounds and sensory attributes of pecans, and their perception by human subjects. Building a sensory-based prediction model would reduce dependency on expensive and time-consuming sensory methods.

Metabolomics and Transcriptomics Analyses Reveal Regulatory Networks Associated with Fatty Acid Accumulation in Pecan Kernels.

Pecans are a globally important tree nut crop. Pecan nuts are rich in fatty acids (FAs), proteins, and flavonoids in addition to thiamine and numerous micronutrients. Although several of these nutriments have been studied in this plant, the comprehensive metabolite variations and molecular mechanisms associated with them have not been fully elucidated. In this study, untargeted metabolomics and transcriptomics were integrated to reveal the metabolite accumulation patterns and their associated molecular mechanisms during pecan kernel development. In total, 4260 (under positive mode) and 2726 (under negative mode) high quality features were retained. Overall, 163 differentially accumulated metabolites were identified. Most components were classified into the categories "organic acids and derivatives" and "lipids and lipid-like molecules." The accumulation patterns of amino acids, FAs, carbohydrates, organic acids, vitamins, flavonoids, and phenylpropanoids alongside embryo development were determined. Furthermore, transcriptomes from four pecan kernel developmental stages were used to assess transcript expression levels. Coexpression analyses were performed between FAs and their related genes. This study provides a comprehensive overview of the metabolic changes and regulations during pecan kernel development. We believe that the identification of nutriment accumulation trends and hub genes associated with the biosynthesis of the components will be valuable for genetically improving this plant.

Using texture analyzer to characterize pecan and olive oil tactile properties, compare to viscometer analysis, and link to fatty acid profile and total polyphenols.

This study aimed to characterize mechanical properties of five pecan oils and one olive oil using a texture analyzer compared to a rotational viscometer; the results were linked to fatty acid profile and total polyphenol content. The seven texture parameters (firmness, consistency, cohesiveness, viscosity index, and stickiness at 5 s, stickiness at 30 s, and delta stickiness) showed significant difference (p ≤.05) among the six oils. Overall, olive oil had higher texture analysis values and significantly higher rotational viscosity than pecan oils. Chemically, C18 fatty acids accounted for approximately 90% of the total fatty acids in the five pecan oils. Olive oil had a higher amount of long-chain, unsaturated fatty acids. Total polyphenols in pecan oils were 8-15 mg gallic acid equivalent (GAE)/100 g, while olive oil contained 27.2 mg GAE/100 g. Correlation analysis demonstrated a significant, positive relationship between "consistency" texture and rotational viscosity measurement. Mechanical properties (seven texture parameters and rotational viscosity) were partially correlated to fatty acid profile, though no universal pattern was identified.

Phytochemical Content and Potential Health Applications of Pecan [Carya illinoinensis (Wangenh) K. Koch] Nutshell.

BACKGROUND: The pecan nutshell contains phytochemicals with various biological activities that are potentially useful in the prevention or treatment of diseases, such as cancer, diabetes, and metabolic imbalances associated with heart diseases. OBJECTIVE: The aim of this study is to update this topic by means of a literature review and include those studies that contribute to the knowledge of the chemical composition and biological activities of pecan nutshell, particularly those related to the therapeutic potential against some chronic degenerative diseases associated with oxidative stress. METHODS: Exhaustive and detailed review of the existing literature was conducted using electronic databases. CONCLUSION: The pecan nutshell is a promising natural product with pharmaceutical uses in various diseases. However, additional research related to the assessment of efficient extraction methods and characterization, particularly the evaluation of the mechanisms of action in new in vivo models, is necessary to confirm these findings and development of new drugs with therapeutic use.

Acute consumption of pecans decreases angiopoietin-like protein-3 in healthy males: a secondary analysis of randomized controlled trials.

Angiopoietin-like proteins (ANGPTL)-3 and -4 regulate lipid metabolism, but the effect of tree nuts of varying fatty acid composition on post-meal responses is unknown. The purpose of the study was to conduct a secondary analysis of two studies on ANGPTL3 and -4 responses to meals containing different tree nuts. We hypothesized that the pecan-containing meal would mitigate postprandial rises in ANGPTL3 compared to the traditional meal without nuts in males, but not females. In addition, we hypothesized that there would be no other differences between any other treatments in ANGPTL3 or -4 responses. The two studies were double-blind, randomized crossover trials. Twenty-two adults (10=male, 12=female) completed study 1, which compared meals containing pecans vs. no nuts (control), and thirty adults (14=male, 16=female) completed study 2, which compared meals containing black walnuts, English walnuts (EW), or no nuts (control). Blood was collected at fasting, 30, 60, 120, and 180min postprandially. In study 1, ANGPTL3 was suppressed more in pecan vs. control in males (iAUC: -579.4±219.4 vs. -128.4±87.1pg/mL/3h, P<.05). In study 2, there was no difference in ANGPTL3 between black walnuts vs. EW, but ANGPTL3 was suppressed more in control vs. black walnuts in females only (iAUC: -196.4±138.4 vs. 102.1±90.1pg/mL/3h, P<.05). There were no differences in ANGPTL4 between treatments. In conclusion, adding pecans to a meal decreased ANGPTL3 in males, but not females. These data highlight the importance of investigating the impact of nutrients and sex on postprandial ANGPTL3 ad -4 responses to better understand their ability to reduce cardiovascular disease risk.

Juglone, a novel activator of ferroptosis, induces cell death in endometrial carcinoma Ishikawa cells.

Ferroptosis is a novel iron-dependent cell death pathway mainly caused by an abnormal redox state and associated with various diseases including cancer. Recently, much attention has been paid to natural compounds that are involved in its activation and inhibition. This is the first ever study to demonstrate the role of juglone isolated from Carya cathayensis green peel in inducing autophagy and inhibiting endometrial cancer (EC) cell migration. Subsequently, Fe2+ accumulation, lipid peroxidation, GSH depletion, the upregulation of HMOX1, and heme degradation to Fe2+ were reported. Juglone was involved in inducing autophagy and inhibiting cell migration and endoplasmic reticulum stress, which are the new hallmarks of cancer treatment. Collectively, our data indicate that juglone as a functional food ingredient induces the programmed cell death of EC cells by activating oxidative stress and suggest a novel therapeutic approach for the treatment and prevention of EC.

Sonication-synergistic natural deep eutectic solvent as a green and efficient approach for extraction of phenolic compounds from peels of Carya cathayensis Sarg.

An excellent high-efficiency natural deep eutectic solvent (NADES, ChCl-MA) was screened out and integrated with pulse-ultrasonication technique for extracting phenolic compounds from Carya cathayensis Sarg. peels (CCSPs). Single factor experiment combined with response surface methodology (RSM) using Box-Behnken design (BBD) were employed to investigate significant factors and optimize their influence on extraction of phenolic compounds. Significant synergistic effect triggered by ChCl-MA based pulse-ultrasonication over other methods used alone were proved by comparative study concerning a variety of bioactive components and antioxidant activities. The second-order kinetic model was developed and validated (R2 > 0.99) to describe the extraction process and its mechanism; and second-order kinetic extraction rate constant (k), saturation concentration (Cs), and initial extraction rate (h) were calculated. FT-IR, DSC and SEM results further demonstrated synergistic effect and influence during extraction. Overall, this study provided a green and high-efficiency alternative for the recovery of various phenolics compounds from plant source by-products.

Carya cathayensis leaf extract attenuates ectopic fat deposition in liver, abdomen and aortic arch in ovariectomized rats fed a high-fat diet.

BACKGROUND: Carya cathayensis1is a commercially cultivated plant in the Zhejiang Province, China. Its nuts exhibit properties of tonifying kidneys and relieving asthma. There have been a few pharmacological studies addressing the function of the leaves of this plant. Our previous studies on C. cathayensis leaf extract (CCE) showed a significant inhibitory effect on weight gain in mice fed a high-fat diet, particularly in female mice. HYPOTHESIS/PURPOSE: To investigate the biological and molecular mechanisms underlying the regulation of ectopic adipose tissue deposition by CCE in ovariectomized rats fed a high-fat diet. STUDY DESIGN: Female Sprague-Dawley rats were ovariectomized and treated with CCE (50, 100, and 200 mg/kg body weight, oral) or estradiol (1 mg/kg body weight, oral) for 8 weeks. METHODS: CCE was subjected to high-performance liquid chromatography to quantify major components. Body weight gain, abdominal fat coefficient, and aortic arch fat coefficient were determined; serum was collected for biochemical analysis; tissues were collected for histopathological examination, quantitative polymerase chain reaction (Q-PCR), and western blotting. RESULTS: The total flavonoid content was determined to be 57.30% in the CCE and comprised chrysin, cardamomin, pinostrobin chalcone, and pinocembrin. Compared with the model group (OVX), CCE treatment reduced body weight gain, abdominal and aortic arch fat coefficients, serum and hepatic lipid profiles, including total cholesterol (TC), total triglycerides (TG), and free fatty acids (FFA) levels; decreased lipid droplets in liver cells; decreased fat accumulation in the aortic arch blood vessel wall and increased its smoothness; decreased the diameter of abdominal fat cells; and reduced serum leptin and adiponectin levels significantly. Serum adiponectin levels significantly correlated with serum TG and hepatic TC levels. Leptin levels positively correlated with serum TG levels and negatively correlated with hepatic TG. Leptin mRNA, peroxisome proliferator-activated receptor (PPARγ) mRNA, and protein expression levels in abdominal adipose tissue were significantly down-regulated. Adiponectin mRNA levels were slightly reduced but not significantly. CONCLUSION: CCE attenuated ectopic fat deposition induced by deficient estrogen and a high-fat diet in rats; this may be associated with activated leptin sensitivity, improved leptin resistance, and regulated adiponectin levels. CCE may improve adipose function to regulate adipocyte differentiation by down-regulating PPARγ. Overall, these results suggest that CCE is a potential phytoestrogen.

Molecular docking simulation and in vitro studies on estrogenic activities of flavonoids from leaves of Carya cathayensis Sarg.

The main purpose of this study was to evaluate the estrogenic properties of total flavonoids (TFs) and five flavonoid monomers (cardamonin (Car), pinostrobin chalcone (PC), wogonin (Wo), chrysin (Chr) and Pinocembrin (PI)) from leaves of Carya cathayensis Sarg (LCC). TFs from LCC were isolated and determined using HPLC. The 3-[4,5-dimethylthiazole-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay and flow cytometry were performed to assess the effects of flavonoids on cell proliferation and cell cycle, respectively. The molecular docking technique was applied to investigate binding conformations of the monomers from LCC to the estrogen receptor ERα and ERß. Gene and protein expression patterns were assessed using quantitative real-time PCR (qRT-PCR) and western blot, respectively. The results showed that TFs, Car, PC, Wo and Chr promoted proliferation of MCF-7 cells and cell transition from the G1 to S phase, and inhabitation of MCF-7 cell proliferation was observed after the treatment of PI. Molecular docking studies confirmed ERs as molecular targets for the monomers. TFs, Car, PC, Wo and Chr from LCC promoted gene expression of ERα, ERß, progesterone receptor (PR) and pS2. Our collective results demonstrated that TFs and monomers from LCC may exert ER agonist activity through competitively bind to ER, inducing ER upregulation and active ER to estrogen response element (ERE)- independent gene regulation. As an abundant natural product, LCC may provide a novel medicinal source for treatment of diseases caused by estrogen deficiency.

A comparison of fatty acid and sensory profiles of raw and roasted pecan cultivars.

Five fatty acids comprise the bulk of the lipid content in pecans: palmitic acid, stearic acid, oleic acid, linoleic acid, and linolenic acid. Understanding the profiles of these fatty acids and how they relate to sensory characteristics may offer an explanation for flavor and flavor defects that may exist in certain cultivars of pecans. The objective of this study was to examine and compare fatty acid profiles of three cultivars of pecans (Major, Lakota, and Chetopa), over two crop years, under raw and roasted preparation methods, and understand the fatty acids association with sensory attributes. Percentages of palmitic, stearic, oleic, linoleic, and linolenic acids to total fatty acid content were determined using gas chromatography, and sensory profiles were generated using descriptive sensory analysis. Similar trends were seen across samples, with oleic acid comprising the majority of the total fatty acids and linolenic acid comprising the smallest percentage. There were significant differences in fatty acid content among cultivars and between pecans in the first and second crop year. Few associations were found between the fatty acids and sensory attributes, which suggest that combinations of the fatty acids contribute to certain pleasant or undesirable flavor attributes in the pecans. Subtle differences in fatty acid composition may lead to variation in flavor and flavor intensity or draw attention to or from certain attributes during consumption. Differences in crop year indicated that fatty acid content and therefore flavor are variable year to year. PRACTICAL APPLICATION: This study will help understand how fatty acid content of pecans varies from year to year. This should be taken into account when manufacturing products with pecans as the nutritional content of the product may change as the result.

Photosystem Disorder Could be the Key Cause for the Formation of Albino Leaf Phenotype in Pecan.

Pecan is one of the most famous nut species in the world. The phenotype of mutants with albino leaves was found in the process of seeding pecan, providing ideal material for the study of the molecular mechanisms leading to the chlorina phenotype in plants. Both chlorophyll a and chlorophyll b contents in albino leaves (ALs) were significantly lower than those in green leaves (GLs). A total of 5171 differentially expression genes (DEGs) were identified in the comparison of ALs vs. GLs using high-throughput transcriptome sequencing; 2216 DEGs (42.85%) were upregulated and 2955 DEGs (57.15%) were downregulated. The expressions of genes related to chlorophyll biosynthesis (HEMA1, encoding glutamyl-tRNA reductase; ChlH, encoding Mg-protoporphyrin IX chelatase (Mg-chelatase) H subunit; CRD, encoding Mg-protoporphyrin IX monomethylester cyclase; POR, encoding protochlorophyllide reductase) in ALs were significantly lower than those in GLs. However, the expressions of genes related to chlorophyll degradation (PAO, encoding pheophorbide a oxygenase) in ALs were significantly higher than those in GLs, indicating that disturbance of chlorophyll a biosynthesis and intensification of chlorophyll degradation lead to the absence of chlorophyll in ALs of pecan. A total of 72 DEGs associated with photosynthesis pathway were identified in ALs compared to GLs, including photosystem I (15), photosystem II (19), cytochrome b6-f complex (3), photosynthetic electron transport (6), F-type ATPase (7), and photosynthesis-antenna proteins (22). Moreover, almost all the genes (68) mapped in the photosynthesis pathway showed decreased expression in ALs compared to GLs, declaring that the photosynthetic system embedded within the thylakoid membrane of chloroplast was disturbed in ALs of pecan. This study provides a theoretical basis for elucidating the molecular mechanism underlying the phenotype of chlorina seedlings of pecan.

Phytosterols extraction from hickory (Carya cathayensis Sarg.) husk with a green direct citric acid hydrolysis extraction method.

This study investigated the direct citric acid hydrolysis extraction method to optimize phytosterols extraction from hickory husk. Single factor experiments followed by a three-level three-factor Box-Behnken experiments were performed. The optimal extraction parameters were determined as: pH of 2.0, liquid-to-solid ratio of 17.12: 1 mL/g, and temperature of 55.81 °C. Practical experiments were carried out in triplicate, and subsequently yielded phytosterols of 912.452 ± 17.452 µg/g DW, in good consistence with the predicted extraction yield of 902.874 µg/g DW. The conductivity of the extract was also found to play effective role under direct citric acid hydrolysis and recorded 36.30 ± 1.08 µs/cm at optimum extraction condition. ß-Sitosterol stigmasterol, campsterol, ergosterol and lupeol were detected as main PSs and triterpenoids in hickory husk using UPLC-Triple-TOF/MS. Finally, the comparison between direct hydrolysis extraction and traditional solvent extraction showed that this new method was more effective and eco-friendlier to extract both free and conjugated phytosterols.

The potential of the pecan nut cake as an ingredient for the food industry.

Pecan nut [Carya illinoinensis (Wangenh.) K. Koch] cake (PNC) is a co-product from the oil extraction industry and its potential as an ingredient for the food industry are not well known. In this work, the nutritional composition and the functional properties of PNC were studied. Additionally, the influence of different solvents (ethanol, water, and acetic acid) on the phytochemical composition and antioxidant capacity (reducing potential of the hydrophilic compounds - RPHC, 2,2-diphenyl-1-picrylhydrazyl - DPPH, and total reducing capacity - TRC) of PNC extracts were established using a simplex-centroid design. PNC is a source of carbohydrates, protein, and dietary fiber (40.5; 21.87 and 13.01 g 100 g-1, respectively). The PNC exhibited a low energy value when compared to the raw nut (398.8 kcal 100 g-1 and 645.54 kcal 100 g-1, respectively). Mg, Mn and Co (416.74; 23.21 mg 100 g-1 and 59.00 µg 100 g-1, respectively) were the main minerals identified in PNC. The PNC also presented functional properties such as emulsifying and oil absorption capacities and a great ability to absorb water. Using the proposed solvent mixture system, the content of total phenolic compounds and condensed tannins recovered from PNC ranged between 172.43 and 2744.24 mg GAE 100 g-1, and 253.42 to 1376.44 mg CE 100 g-1, respectively. The antioxidant capacity of the PNC extract was showed through its ability to reduce hydrophilic (172.06-1714.96 mg GAE 100 g-1) to transfer hydrogen atoms (12.55-74.11% scavenging activity) and lipophilic compounds (509.87-2070.80 mg QE 100 g-1) using RPHC, DPPH, and TRC methods, respectively. Combining ethanol, water, and acetic acid at 30 °C for 15 min, positively affects the extraction of bioactive compounds from PNC, as well as the antioxidant activity of the extracts. The physicochemical, functional, phytochemical, and antioxidant properties demonstrate that pecan nut cake may represent a potential ingredient or additive for the food, pharmaceutical, and cosmetic industries.

Cellular evaluation of the antioxidant activity of U.S. Pecans [Carya illinoinensis (Wangenh.) K. Koch].

Clinical trials show an inverse relationship between the consumption of antioxidant-rich tree nuts and the development of chronic diseases. This study examined antioxidant efficacy of U.S. pecans using a modified cellular antioxidant activity (CAA) assay with comparisons to data from in vitro antioxidant assays (hydrophilic-oxygen radical absorbance capacity {H-ORACFL} and ferric reducing antioxidant power {FRAP}). Crude phenolic extracts from both raw and roasted pecans were analyzed. In the CAA assay, pecan phenolics were taken up by human colorectal adenocarcinoma (Caco-2) cells and bestowed CAA, determined by monitoring the fluorescence of 2',7'-dichlorofluorescein. Phenolics (25-100 µg/mL) demonstrated a reduction in fluorescence by 37-69% for raw and 26-68% for roasted pecans. The primary active phenolic constituents were determined by high-performance liquid chromatography-electrospray ionization-mass spectrometry (HPLC-ESI-MS) to be epi(catechin) dimers and trimers. These oligomeric procyanidins, ranging in size from 560 to 840 g/mol appear to be small enough for cellular uptake, showing pecans are an effective antioxidant in biological systems, regardless of roasting.

Mechanism of Juglone-Induced Cell Cycle Arrest and Apoptosis in Ishikawa Human Endometrial Cancer Cells.

The molecular mechanism of Juglone-induced cell cycle arrest and apoptosis in human endometrial cancer cells was investigated. Juglone was purified from the green husk of Carya cathayensis Sarg and identified by HPLC, LC-MS/MS, and NMR. At an IC50 of 20.81 µM, juglone significantly inhibited Ishikawa cell proliferation, as shown by S phase arrest mediated by inactivation of cyclin A protein ( p < 0.05). The ROS levels increased significantly after exposure to juglone, which paralleled increases in the mRNA and protein expression of p21 and decreases in the levels of CDK2, cdc25A, CHK1, and cyclin A. The expression of Bcl-2 and Bcl-xL was significantly down-regulated, whereas the expression of Bax, Bad and cyto c was up-regulated, and we later confirmed the involvement of the mitochondrial pathway in juglone-induced apoptosis. Our in vitro results stated that juglone can be studied further as an effective natural anticancer agent.

Total flavonoids from the Carya cathayensis Sarg. leaves inhibit HUVEC senescence through the miR-34a/SIRT1 pathway.

Aging shows a significant relationship with changed vascular structure and function, and advancing age is a major nonmodifiable risk factor in the occurrence of cardiovascular diseases. The senescence of endothelial cells is one of the hallmarks of vascular aging and can induce vascular dysfunction. This study aimed to investigate the effect of total flavonoids (TFs) on human umbilical vein endothelial cells (HUVEC) senescence and identify the potential mechanisms involved. A HUVEC senescence model was induced by angiotensin II. The senescence markers, including senescence-associated ß-galactosidase (SA-ß-gal), p53, p21, and stagnate G0/G1, were measured. The effects of TFs on miR-34/ SIRT1 were examined by quantitative polymerase chain reaction analysis and Western blot analysis. TFs decreased the percentage of SA-ß-gal-positive cells and resulted in G0/G1 cell cycle arrest, while the percentage of cells in the S phase increased. Furthermore, TFs reduced miR-34a expression and increased the expression of SIRT1. After treatment with TFs and a miR-34a inhibitor, the percentage of SA-ß-gal-positive cells and the expression of miR-34a decreased, and the expression of SIRT1 increased. The TFs inhibited HUVEC senescence, and the mechanism was related to the miR-34a/Sirtuin1 pathway.