Ellagic Acid Content Research Articles

Chemical Characterization, Antilipidemic Effect and Anti-Obesity Activity of Ludwigia octovalvis in a Murine Model of Metabolic Syndrome.

Ludwigia octovalvis (Jacq.) P.H. Raven is widely used in traditional medicine for different illnesses, including diabetes and hypertension. However, its impact on lipotoxicity and metabolic syndrome in vivo has not been addressed. Therefore, the aim of this study was to evaluate the effects of this plant on the metabolic syndrome parameters in a C57BL6J mouse hypercaloric diet model. L. octovalvis hydroalcoholic extract and its ethyl acetate fraction (25 mg/kg/day) were used for sub-chronic assessment (10 weeks). Additionally, four subfractions (25 mg/kg) were evaluated in the postprandial triglyceridemia test in healthy C57BL6J mice. The hydroalcoholic extract and ethyl acetate fraction significantly decreased body weight gain (-6.9 g and -1.5 g), fasting glycemia (-46.1 and -31.2 mg/dL), systolic (-26.0 and -22.5 mmHg) and diastolic (-8.1 and 16.2 mmHg) blood pressure, free fatty acid concentration (-13.8 and -8.0 μg/mL) and insulin-resistance (measured by TyG index, -0.207 and -0.18), compared to the negative control. A postprandial triglyceridemia test showed that the effects in the sub-chronic model are due, at least in part, to improvement in this parameter. L. octovalvis treatments, particularly the hydroalcoholic extract, improve MS alterations and decrease free fatty acid concentration. These effects are possibly due to high contents of corilagin and ellagic acid.

Enhanced Anticancer Effect of Unripe Rubus occidentalis on Gastric Cancer Cells

Background Black raspberry ( Rubus occidentalis L.) has various bioactive properties, including antioxidant, anti-inflammatory, and anticancer activities. Recently, it has been proposed that the pharmacological efficacy of black raspberry varies with its maturation stage. In the current study, we explored the relationship between maturation stage and anticancer activity in gastric cancer cell lines. Materials and Methods Black raspberry extracts were prepared from both ripe samples 41 days after the fruit set and unripe samples 22 days after the fruit set. Their anticancer activities were compared by assessing the effects on proliferation, apoptosis, invasiveness, and clonogenicity in gastric cancer cell lines. Results Both the ripe and unripe black raspberry samples were effective in inhibiting cell proliferation, invasion, and clonogenic activity. However, the unripe black raspberry extract showed a stronger effect on these activities compared to ripe black raspberry extract. Additionally, the unripe fruit extract increased apoptotic body formation, caspase-3 activity, Bak expression and decreased Bcl2 expression. Furthermore, the unripe black raspberry sample was found to contain higher contents of ellagic acid and gallic acid, and had a stronger suppressive effect on p-ERK and p-AKT protein levels compared to the ripe black raspberry sample. Conclusion Unripe black raspberry has greater anti-cancer effects than fully ripe black raspberry in gastric cancer cells.

Micronized Powder of Raspberry Pomace as a Source of Bioactive Compounds.

Red raspberries, which contain a variety of nutrients and phytochemicals that are beneficial for human health, can be utilized as a raw material in the creation of several supplements. This research suggests micronized powder of raspberry pomace production. The molecular characteristics (FTIR), sugar, and biological potential (phenolic compounds and antioxidant activity) of micronized raspberry powders were investigated. FTIR spectroscopy results revealed spectral changes in the ranges with maxima at ~1720, 1635, and 1326, as well as intensity changes in practically the entire spectral range analyzed. The discrepancies clearly indicate that the micronization of the raspberry byproduct samples cleaved the intramolecular hydrogen bonds in the polysaccharides present in the samples, thus increasing the respective content of simple saccharides. In comparison to the control powders, more glucose and fructose were recovered from the micronized samples of the raspberry powders. The study's micronized powders were found to contain nine different types of phenolic compounds, including rutin, elagic acid derivatives, cyanidin-3-sophoroside, cyanidin-3-(2-glucosylrutinoside), cyanidin-3-rutinoside, pelargonidin-3-rutinoside, and elagic acid derivatives. Significantly higher concentrations of ellagic acid and ellagic acid derivatives and rutin were found in the micronized samples than in the control sample. The antioxidant potential assessed by ABTS and FRAP significantly increased following the micronization procedure.

Assessing the anti-collagenase activity and phytochemical evaluation of eight plants cultivated in Egypt; developing a validation method for the ellagic acid content in Chrysophyllum oliviforme using HPLC-DAD

Preventing aging signs is a vast cosmetic quest hence; the authors felt the necessity to focus on emerging new plant extracts by the evaluation of anti-aging potential of eight plants cultivated in Egypt. Total phenolics (TPC), total flavonoids content (TFC) and collagenase assay were performed; only four plants were subjected to (ORAC) assay, ferrozine iron metal chelation assay and (HPLC) analysis against polyphenolic standard; validation method according to ICH guidelines for ellagic acid content in C. oliviforme by HPLC-DAD was performed, molecular docking simulation was implemented by (MOE) module. C. oliviforme exhibited the highest anti-collagenase with the lowest (IC50), (TPC = 299.70 ± 16.97 mg/GAE), method of validation of C. oliviforme extract following the ICH guidelines of ellagic acid content (147.446 ± 0.00041 mg/g); C. oliviforme was the most potent extract and was standardized to be reproducible for the industrial scale.

Validation and standardization of Gallic acid and Ellagic acid in Quercus infectoria, Terminalia chebula and Pistacia integerrima.

Due to its medicinal properties, Pistacia integerrima is in high demand and is extensively used as a key ingredient in various formulations. However, its popularity has led to IUCN threatened category list. In Ayurvedic texts, such as Bhaishajaya Ratnavali, Quercus infectoria is recommended as a substitute for P. integerrima in different formulations. Additionally, Yogratnakar highlights that Terminalia chebula shares similar therapeutic properties with P. integerrima. The objective of the current study was to gather scientific data on metabolite profiling and markers-based comparative analysis of Q. infectoria, T. chebula and P. integerrima. In present study, hydro-alcoholic and aqueous extracts of all three plants were prepared and standardized for the comparative evaluation of secondary metabolites. Thin-layer chromatography was carried out for the comparative fingerprinting of the extracts using chloroform: methanol: glacial acetic acid: water (60: 8: 32: 10, v/v/v/v) as a solvent system. A fast, sensitive, selective, and robust HPLC method was developed to determine gallic acid and ellagic acid from both extracts of all three plants. The method was validated for precision, robustness, accuracy, limit of detection and quantitation as per the International Conference on Harmonization guidelines. The TLC analysis revealed the presence of several metabolites, and the pattern of metabolites in the plants exhibited a certain degree of similarity. A highly precise and reliable quantification technique was created for gallic acid and ellagic acid, operating within the linear concentration range of 81.18-288.22 µg/mL and 3.83-13.66 µg/mL, respectively. The correlation coefficients for gallic acid and ellagic acid were 0.999 and 0.996, indicating strong relationships. The gallic acid content in all three plants ranged from 3.74% to 10.16% w/w, while the ellagic acid content ranged from 0.10% to 1.24% w/w. This pioneering scientific approach highlights the phytochemical similarities between Q. infectoria, T. chebula and P. integerrima.

Effects of Different Nitrogen Forms on Blackberry Fruit Quality.

To study the optimal form of nitrogen (N) application and to determine the best harvest date for blackberries, different N fertilizers were applied during the critical growth period of blackberry plants. The results showed that NH4+-N significantly improved the appearance of blackberry fruits, including their size, firmness, and color, and promoted the accumulation of soluble solids, sugars, anthocyanin, ellagic acid, and vitamin C (VC), while fruit treated with NO3--N accumulated more flavonoids and organic acids and had improved antioxidant capacity. In addition, the fruit size, firmness, and color brightness decreased with the harvest period. While the contents of sugars, anthocyanin, ellagic acid, flavonoids, and VC were higher in the early harvests and then decreased as the season progressed, the total antioxidant capacity and DPPH radical scavenging capacity increased. In all, application of NH4+-N is recommended, as it is more beneficial to fruit appearance, taste, and nutritional quality. Harvests in the early stage help to obtain a good fruit appearance, while harvests in the middle and later stages are more beneficial to fruit taste and quality. This study may help growers to determine the best fertilization scheme for blackberries and choose the appropriate harvest time according to their needs.

Extraction, phytochemical characterization and anti-cancer mechanism of Haritaki churna: An ayurvedic formulation.

Haritaki churna (HC), a single herb ayurvedic formulations is known to be prescribed for various gastro-intestinal disorders in Ayurveda. Haritaki churna aqueous extract (HCAE) has anti-cancer activity against different types of cancer cells with an IC50 in the range of 50-97 μg/ml. Bioavailability of Haritaki Churna is very high in digestive track and treatment of colorectal cancer cells HCT-116, DLD1, HT-29 with HCAE reduces its cellular viability with anti-cancer IC50 70μg/ml. HCAE consumption is safe for human as it didn't affect the cellular viability of primary human PBMCs or non-cancerogenic HEK-293 cells. Haritaki churna was found to be stable in biological gastric fluids and bioactive agents are not losing their anti-cancer activity under such harsh conditions. The HPLC Chromatogram of HCAE is giving 13 major peaks and 11 minor peaks. Exploiting LC-MS, IR and NMR spectroscopic techniques, a total of 13 compounds were identified from HCAE namely Shikimic acid, Chebulic acid, gallic acid, 5-hydroxymethylfurfural, Protocatechuic acid, 4-O-galloyl-shikimic Acid, 5-O-galloyl-shikimic Acid, Methylgallate, corilagin, 1, 2, 6, Tri-O-galloyl β-D-glucose, chebulagic acid, chebulinic acid, and Ellagic acid. Reconstitution and subtraction of phytochemicals from the mixture indicate that Ellagic acid significantly contribute into anti-cancer effect of HCAE. Cancer cells treated with ellagic acid from HCAE were incapable of completing their cell-cycle and halted the cell-cycle at DNA synthesis S-Phase, as demonstrated by decreased cyclin A2 expression levels with increasing ellagic acid concentration. Halting of cells at S-phase causes induction of apoptosis in cancer cells. Cancer cells exhibiting DNA fragmentation, changes in expression of several apoptotic proteins such as Bcl2, cytochrome-c and formation of cleaved products of caspase 3 and PARP-1 suggests ellagic acid induces cell death via mitochondrial pathway of apoptosis.

Physiological and metabolomic analyses reveal the effects of different NH4+:NO3− ratios on blackberry fruit quality

Blackberries are important economic fruits, and they not only have a distinctive flavor but are also rich in nutritional and medicinal value. With the aim of elucidating the effects of different NH4+:NO3− ratios on blackberry fruit quality, the physiological characteristics and metabolomics of blackberry fruits were investigated in this study. The results revealed that the NH4+:NO3−/50:50 ratio treatment showed the maximum values for single fruit weight, transverse diameter, longitudinal diameter and the solidity–acid ratio of blackberry fruit as well as the maximum flavonoid and ellagic acid contents. The NH4+:NO3−/75:25 ratio treatment exhibited the maximum fructose, sucrose, glucose and anthocyanin contents and the highest superoxide dismutase (SOD) and catalase (CAT) activities. Moreover, a total of 857 known metabolites were detected in the blackberry fruits. Interestingly, NH4+ could increase the levels of amino acids and metabolites belonging to the indole and derivative classes in blackberry fruits, and D-proline, cysteine, tryptophan and indole may play vital roles in the formation of blackberry fruit quality. To produce blackberry fruit that is larger and has better flavor, a 50:50 ratio of NH4+:NO3−is recommended, while when the goal is higher anthocyanin and amino acid content, a 75:25 ratio of NH4+:NO3− is more appropriate. In addition, a 100:0 ratio of NH4+:NO3− is best for fruit storge. This study serves as a reference and provides technical support for the application of N to improve fruit quality and antioxidant capacity during small berries cultivation.

Optimization of ellagic acid extraction from blueberry pulp through enzymatic hydrolysis combined with ultrasound-assisted organic solvent

The aim of this study was to optimize the technology used to extract ellagic acid from blueberry pomace. For single factor analysis, an enzymatic hydrolysis test showed that temperature, pH value and liquid-to-material ratio exhibited significant effects on ellagic acid content, as did three factors (the liquid-to-material ratio, ultrasonication time and temperature) for the ultrasound-assisted organic solvent method. A Box–Behnken design was applied for the enzymatic hydrolysis test and the ultrasound-assisted organic solvent method to determine the optimal combination scheme. For enzymatic hydrolysis, this scheme was 7:1 for the liquid-to-material ratio, 46 °C for the temperature of enzymatic hydrolysis and 4.9 for the pH value; under these conditions, an ellagic acid content of 1.96 mg/g (FW) was obtained. The optimal combination scheme for the ultrasound-assisted organic solvent was 2:1 for the liquid-to-material ratio, 65 °C for ultrasonication temperature and 82 min for the ultrasonication time, which produced ellagic acid contents of 7.92 mg/g (FW) and an extraction yield of 32.41%, respectively. The content and yield of ellagic acid in the extraction from blueberry pomace were greatly increased by combining enzymatic hydrolysis and an ultrasound-assisted organic solvent method. This study provides a theoretical reference for utilizing and developing blueberry pomace.

The phenolics, antioxidant activity and in vitro digestion of pomegranate (Punica granatum L.) peels: an investigation of steam explosion pre-treatment.

Pomegranate peels, the main byproduct of pomegranate production, are rich in phenolic compounds that are known for their effective antioxidant properties and have vast application prospects. In this study, steam explosion, an environmentally friendly technique, was applied to pretreat pomegranate peels for phenol extraction. We investigated the effects of explosion pressure, duration, and particle size on the content of total and individual phenolics, and antioxidant activity of pomegranate peels before and after in vitro digestion. The optimal conditions for a steam explosion for pomegranate peels in terms of total phenol content were a pressure of 1.5 MPa, a maintenance time of 90 s, and a particle size of 40 mesh. Under these conditions, pomegranate peel extract presented a higher yield of total phenols, gallic acid, and ellagic acid. However, it also had a lower content of punicalin and punicalagin, compared to the unexploded peels. There was no improvement in the antioxidant activity of pomegranate peels after the steam explosion. Moreover, the content of total phenol, gallic acid, ellagic acid, punicalin, and punicalagin, as well as the antioxidant activity of pomegranate peels, all increased after gastric digestion. Nevertheless, there was a large variation in the pomegranate peel processed by different pressure, duration, and sieve fractions. Overall, this study demonstrated that steam explosion pre-treatment could be an efficient method for improving the release of phenolics, especially gallic acid, and ellagic acid, from pomegranate peels.

Variation in Phenolic Compounds, Antioxidant and Antibacterial Activities of Extracts from Different Plant Organs of Meadowsweet (Filipendula ulmaria (L.) Maxim.).

Meadowsweet (Filipendula ulmaria (L.) Maxim.) has been widely used in the treatment of various diseases. The pharmacological properties of meadowsweet are derived from the presence of phenolic compounds of a diverse structure in sufficiently large quantities. The purpose of this study was to examine the vertical distribution of individual groups of phenolic compounds (total phenolics, flavonoids, hydroxycinnamic acids, catechins, proanthocyanidins, and tannins) and individual phenolic compounds in meadowsweet and to determine the antioxidant and antibacterial activity of extracts from various meadowsweet organs. It was found that the leaves, flowers, fruits, and roots of meadowsweet are characterized by a high total phenolics content (up to 65 mg g-1). A high content of flavonoids was determined in the upper leaves and flowers (117-167 mg g-1), with high contents of hydroxycinnamic acids in the upper leaves, flowers, and fruits (6.4-7.8 mg g-1); high contents of catechins and proanthocyanidins in the roots (45.1 and 3.4 mg g-1, respectively); and high tannin content in the fruits (38.3 mg g-1). Analysis of extracts by high-performance liquid chromatography (HPLC) showed that the qualitative and quantitative composition of individual phenolic compounds in various parts of the meadowsweet varied greatly. Among the flavonoids identified in meadowsweet, quercetin derivatives dominate, namely quercetin 3-O-rutinoside, quercetin 3-β-d-glucoside, and quercetin 4'-O-glucoside. Quercetin 4'-O-glucoside (spiraeoside) was found only in the flowers and fruits. Catechin was identified in the leaves and roots of meadowsweet. The distribution of phenolic acids across the plant was also uneven. In the upper leaves, a higher content of chlorogenic acid was determined, and in the lower leaves, a higher content of ellagic acid determined. In flowers and fruits, a higher contents of gallic, caftaric, ellagic, and salicylic acids were noted. Ellagic and salicylic acids were also dominant among phenolic acids in the roots. Based on the results of the analysis of antioxidant activity in terms of the ability to utilize the radicals of 2,2-diphenyl-1-picrylhydrazine (DPPH) and 2,2'-azino-bis(3-ethylbenzthiazolino-6-sulfonic acid) (ABTS) and in terms of iron-reducing ability (FRAP), the upper leaves, flowers, and fruits of meadowsweet can be considered plant raw materials suitable to obtain extracts with high antioxidant activity. Extracts of plant fruits and flowers also showed high antibacterial activity against the bacteria Bacillus subtilis and Pseudomonas aeruginosa.

Polyphenols in Jabuticaba (Plinia spp.) Peel Flours: Extraction and Comparative Evaluation of FTIR and HPLC for Quantification of Individual Compounds

Jabuticabas are wild fruits native to Brazil, and their peels, the main residue from jabuticaba processing, contain significant amounts of bioactive compounds, which are mostly phenolics. Conventional methods based on the estimation of total extractable phenolics (TEP-Folin-Ciocalteau) or total monomeric anthocyanins (TMA) have limitations and may not reflect the actual antioxidant potential of these peels. Analytical methods, such as high-performance liquid chromatography (HPLC), are more appropriate for the quantification of specific phenolics, and can be used as a reference for the construction of mathematical models in order to predict the amount of compounds using simple spectroscopic analysis, such as Fourier Transform Infrared Spectroscopy (FTIR). Therefore, the objectives of this study were (i) to evaluate the composition of specific polyphenols in flours prepared from jabuticaba peels and verify their correlation with TEP and TMA results from a previous study, and (ii) to employ FTIR coupled with chemometrics to predict the concentrations of these polyphenols in jabuticaba peel flours (JPFs) using HPLC as a reference method. Cyanidin-3-glucoside (C3G), ellagic acid (EA) and delphinidin-3-glucoside (D3G) were the main polyphenols found in the samples. The C3G contents ranged from 352.33 mg/100 g (S10) to 1008.73 mg/100 g (S22), with a strong correlation to TMA (r = 0.97; p = 0.00) and a moderate correlation to TEP (r = 0.45; p = 0.02). EA contents ranged from 163.65 mg/100 g (S23) to 334.69 mg/100 g (S11), with a moderate to strong correlation to TEP (r = 0.69; p = 0.00). The D3G values ranged from 94.99 mg/100 g (S10) to 203.36 mg/100 g (S5), with strong correlations to TMA (r = 0.91; p = 0.00) and C3G levels (r = 0.92; p = 0.00). The developed partial least squares-PLS models based on FTIR data provided satisfactory predictions of C3G and EA levels, reasonably matching those of HPLC.

Optimization of ultrasound-assisted deep eutectic solvent extraction of bioactive compounds from pomegranate peel using response surface methodology.

The efficient extraction of polyphenols from pomegranate peels using a deep eutectic solvent (DES) and ultrasound-assisted extraction (UAE) was investigated. A Box-Behnken design was used to investigate the effects of four independent variables (water content, liquid-to-solid ratio, ultrasonic power, and extraction time) on total polyphenol content (TPC), punicalagin content (PC), and ellagic acid content (EC). Optimized DES-based UAE conditions were as follows: TPC (water content, 29.30%; liquid-to-solid ratio, 53.50 mL/g; ultrasonic power, 238.20W; extraction time, 29.50min), PC (water content, 25.65%; liquid-to-solid ratio, 44.20 mL/g; ultrasonic power, 120W; extraction time, 20min), and EC (water content, 33.13%; liquid-to-solid ratio, 60 mL/g; ultrasonic power, 300W; extraction time, 20min). Under these optimal conditions, the experimental values for TPC, PC, and EC were 67.50mg GAE/g, 130.65mg/g, and 2.04mg/g, respectively; these values were consistent with the predicted values.

Post-secretory synthesis of a natural analog of iron-gall ink in the black nectar of Melianthus spp.

The black nectar produced by Melianthus flowers is thought to serve as a visual attractant to bird pollinators, but the chemical identity and synthesis of the black pigment are unknown. A combination of analytical biochemistry, transcriptomics, proteomics, and enzyme assays was used to identify the pigment that gives Melianthus nectar its black color and how it is synthesized. Visual modeling of pollinators was also used to infer a potential function of the black coloration. High concentrations of ellagic acid and iron give the nectar its dark black color, which can be recapitulated through synthetic solutions containing only ellagic acid and iron(III). The nectar also contains a peroxidase that oxidizes gallic acid to form ellagic acid. In vitro reactions containing the nectar peroxidase, gallic acid, hydrogen peroxide, and iron(III) fully recreate the black color of the nectar. Visual modeling indicates that the black color is highly conspicuous to avian pollinators within the context of the flower. Melianthus nectar contains a natural analog of iron-gall ink, which humans have used since at least medieval times. This pigment is derived from an ellagic acid-Fe complex synthesized in the nectar and is likely involved in the attraction of passerine pollinators endemic to southern Africa.

Bioactive Properties of Kakadu Plum-Blended Products.

Kakadu plum (Terminalia ferdinandiana), endemic to Australia, is growing in popularity due to its high levels of vitamin C and strong antioxidant properties. In this study, Kakadu plum fruit powder was used as a functional food ingredient with other plant materials to develop value-added products to enhance their nutritional and commercial value. The present study determined the bioactive properties of nine products, including three Kakadu plum fruit powder samples produced from different processing batches and five Kakadu plum-blended products. Vitamin C, the total phenolic content, and the ellagic acid content were determined. Bioactive properties such as antioxidant, antidiabetic, and antimicrobial assays were also performed. Cytotoxicity was tested to obtain more specific product information regarding food safety. Kakadu plum-blended products showed lower cytotoxicity and lower bioactive properties (antioxidant and antibacterial activities) in comparison to Kakadu plum powder. However, overall, most of the bioactive properties were shown to be higher in the blends when compared with the commercial blueberry powder as a benchmark antioxidant product. Therefore, there is great potential for Kakadu plum to contribute to the growing functional food and ingredient markets.

Selective determination of ellagic acid in aqueous solution using blue-green emissive copper nanoclusters

Development of beneficial sensors to analyze ellagic acid concentrations is of great importance for food safety and human health. Herein, a facile and fast fluorescent probe was carried out for the excellently selective and sensitive measurement of ellagic acid in real samples through histidine protected copper nanoclusters (histidine@Cu NCs) as a nanosensor. This as-developed histidine@Cu NCs were performed through UV–vis absorption spectroscopy, fluorescence spectroscopy, transmission electron microscopy, Fourier-transform infrared spectroscopy, X-ray photoelectron spectroscopy and fluorescence lifetime analysis. The TEM image revealed that this nanomaterial had spherical features with the average diameter of 2.5 ± 0.05 nm. The blue-green fluorescence of this Cu NCs was found under the UV light. Meanwhile, the maximum excitation and emission wavelength were located at 387 nm and 488 nm. After addition of ellagic acid, the fluorescence of histidine@Cu NCs was slowly weakened with excellent linear range of 0.5–300 μM and detection limit of 0.077 μM. The fluorescence weakening mechanism of this nanosensor were attributed to the inner filter effect (IFE) and static quenching. Finally, this as-established analysis platform was successfully employed to measure ellagic acid in real samples.

Jaboticaba (Myrciaria cauliflora) Fruit Extract Suppressed Aberrant Crypt Formation in 1,2-Dimetylhydrazine-Induced Rats.

Early intervention can significantly improve the colorectal cancer survival rate. Foods rich in phenolic compounds, such as jaboticaba (Myrciaria cauliflora), may prevent tumorigenesis. We investigated the effectivity of jaboticaba whole fruit ethanolic extract (FEX) in suppressing aberrant crypt foci (ACF), the earliest lesion of colorectal cancer (CRC), in 1,2-dimethylhydrazine (DMH)-induced rats and the underlying mechanisms related to the gut microbiota composition and short chain fatty acid (SCFA). This study was approved by the Institutional Animal Care and Use Committee (IACUC) of Providence University (Trial Registration Number 20180419A01, registration date: 22 December 2018). The FEX contains gallic acid and an especially high ellagic acid concentration of 54.41 ± 1.80 and 209.79 ± 2.49mg/100g FEX. The highest total ACF number (150.00 ± 43.86) was recorded in the DMH control (D) group. After 56 days of oral FEX treatment, the total ACF number in the low FEX dosage (DL) group was significantly lower compared to the D group (p < 0.05). The large-sized ACF (> 5 foci), which has a higher probability of progressing to later stage, was significantly decreased in the high FEX dosage (DH) group. The 16s rDNA metagenomic sequencing of the cecal material revealed that the CRC biomarker Lachnoclostridium was significantly suppressed in the DH group (p < 0.05), whereas some SCFA-producing taxa and the cecal butyrate concentration were significantly elevated in the DL and DH groups (p < 0.05). This study demonstrated the potential of jaboticaba whole fruit in CRC prevention, especially in the initial stage, by shifting gut microbiota composition and improving cecal butyrate level.

Effects of nitrogen application level on the physiological characteristics, yield and fruit quality of blackberry

To study the optimal amount of nitrogen application for blackberry plants, different amounts of ammonium sulfate were applied to three-year-old healthy ‘Shuofeng’ blackberry plants at three key growth stages (early bud burst, the flowering stage and the fruit veraison stage) to explore the effects of nitrogen fertilizer application level on the physiological characteristics of leaves, yield and fruit quality of blackberry. Overall, the leaves turned yellow, and cellular metabolism was disrupted in the CK group (no nitrogen fertilization). Among the nitrogen treatments, the leaf chlorophyll content, photosynthesis efficiency and antioxidant enzyme activity were significantly higher under the T2 (40 g plant−1) and T3 (60 g plant−1) treatments than under the T1 (20 g plant−1) and T4 (80 g plant−1) treatments, while the contents of ascorbic acid, glutathione, hydrogen peroxide and malondialdehyde showed the opposite trends. The yield, single-fruit weight, solids–acids ratio and anthocyanin contents of blackberry fruits increased first and then decreased with increasing nitrogen application. The contents of polyphenols and ellagic acid in fruits decreased with increasing nitrogen application. Correlation and principal component analysis showed that yield and its components were closely related to leaf photosynthesis characteristics and the antioxidant system (P<0.05). Application of 20 g ammonium sulfate per plant at early bud burst, the flowering stage and the fruit veraison stage was profitable, which resulted in more vigorously growth, increased yield and fruit quality, while the production cost was the most economical because of the low fertilizer consumption.

Addition of ellagic acid improved the immune ability and delayed the apoptosis of ovarian granulosa cells of Guizhou black goat

Context Follicular development plays an important role in the growth and reproduction of female mammals. Ellagic acid (EA), as a natural antioxidant, has been used in freezing protection of pig semen. However, the effects of EA on immunity and the anti-apoptotic ability of ovarian granulosa cells (GCs) are still unclear. Aims The aim of this study was to analyse the effects of different concentrations of EA on the immune and anti-apoptotic ability of ovarian GCs of Guizhou black goats. Methods In this study, different concentrations of EA (0, 50, 100, 150, 200 μmol/L) were added to the culture of ovarian GCs in vitro, and Cell-Counting Kit 8 (CCK8) assay, cell wound scratch assay, and real-time fluorescence quantitative polymerase chain reaction (RT–qPCR) assay were used to detect the effects of different concentrations of EA on the proliferation, migration, and reproductive marker genes of ovarian GCs. Then the optimal addition concentration of EA was selected and the effects of EA supplementation on immune factors, cytochrome P450 family 19 subfamily A member 1 gene (CYP19A1), estradiol concentrations, intracellular reactive oxygen species concentrations, and apoptosis-related protein expression were detected by RT–qPCR, enzyme-linked immunosorbent assay (ELISA), ROS, and western blotting on the basis of the optimal addition concentration. Key results The CCK8 test and cell scratch test showed that the addition of EA could significantly inhibit the proliferation and migration ability of ovarian GCs compared with the control group, and a dose effect was observed with the increase in concentration. RT–qPCR results showed that different concentrations of EA significantly increased the expression of genes associated with reproduction, including bone morphogenetic protein 15 (BMP15), bone morphogenetic protein receptor 1B (BMPR-1B), growth differentiation fFactor 9 (GDF9), and follicle-stimulating hormone β subunit (FSHβ), and the maximum increase was observed at 150 μmol/L EA. Further analyses using 150 μmol/L EA as the optimal concentration showed significantly increased expressions of CYP19A1, interleukin-10 (IL-10), and superoxide dismutase (SOD2) after EA supplementation, while the expression of IL-8 was significantly decreased compared with those of the control group. ELISA and ROS showed that both intracellular and extracellular estradiol concentrations were higher, while ROS concentrations were significantly lower than those in the control group. Western blotting results showed that 150 μmol/L EA significantly decreased the expression of Caspase-3 and Caspase-9 and the ratio of BCL2-associated X:B-cell lymphoma-2. Conclusions The supplementation of 150 μmol/L EA had significant effects on improving GC immunity and delaying GC apoptosis in goats. The addition of EA also increased the expression of BMP15, BMPR-1B, GDF9, FSHβ, and CYP19A1 and promoted the secretion of estradiol in GCs. Implications These results provided a preliminary lead for further research on the effect of EA on the maturation and development of goat oocytes in vitro.

Absorption and Metabolism of Urolithin A and Ellagic Acid in Mice and Their Cytotoxicity in Human Colorectal Cancer Cells.

Ellagic acid is a natural polyphenol compound found in pomegranates, walnuts, and many berries. It is not easily absorbed, but it could be metabolized to urolithins by the gut microbiota. Urolithin A, one of the ellagic acid metabolites, has been proved to prolong the lifespan of C. elegans and increases muscle function of mice. The purpose of this current study was to analyze the absorption and metabolites of urolithin A and ellagic acid in mice and the anticancer effects of urolithin A, urolithin B, and ellagic acid in colorectal cancer cells. Urolithin A and urolithin B were synthesized and analyzed by HPLC and NMR. A pharmacokinetic study of urolithin A was performed in mice by analyzing urolithin A and its metabolites in urines. Absorption and biotransformation of ellagic acid were also studied in mice by analyzing the plasma, liver, and feces. The cytotoxicity of urolithin A, urolithin B, and ellagic acid was assayed in SW480, SW620, HCT 116, and HT-29 cells. Urolithin A and urolithin B were synthesized and purified to reach 98.1% and 99% purity, respectively, and the structures were identified by NMR. In urolithin A intake analysis, urolithin A was only detectable at 3 h, not at 6-24 h; it suggested that urolithin A was rapidly metabolized to some unknown metabolites. Using UPLC-MS/MS analysis, the metabolites might be urolithin A 3-O-glucuronide, urolithin A 3-sulfate, and urolithin A-sulfate glucuronide. After feeding mice with ellagic acid for consecutive 14 days, ellagic acid contents could be detected in the fecal samples, but not in plasma and liver, and urolithin A was not detected in all samples. It suggests that ellagic acid is not easily absorbed and that the biotransformation of ellagic acid to urolithin A by intestinal flora might be very low. From the cytotoxicity assay, it was found that there was anticancer effect in urolithin A and urolithin B but not in ellagic acid. In contrast, ellagic acid promoted the proliferation of SW480 and SW620 cells.