Amelioration activity of the high bioaccessible chrysanthemum (Gongju) phenolics on alcohol-induced oxidative injury in AML-12 cells.

The main bioavailable phenolics from of Gongju (GJ) and their mechanism for hepato-protection remain unclear. To select the GJ phenolics with high bioavailability, chrysanthemum digestion and Caco-2 cells were used and their hepato-protective potential were examined by using AML-12 cells. The digestive recovery and small intestinal transit rate of the main phenolic compounds ranged from 28.52 to 69.53% and 6.57% âˆ¼ 15.50%, respectively. Among them, chlorogenic acid, 3,5-dicaffeoylquinic acid, and 1,5-dicaffeoylquinic acid, showed higher small intestinal transit rates and digestive recoveries. Furthermore, we found that by increasing intracellular Catalase (CAT) and Superoxide dismutase (SOD) viability and lowering Malondialdehyde (MDA) level (P < 0.05), 3,5-dicaffeoylquinic acid significantly mitigated the oxidative damage of AML-12 liver cells more than the other two phenolics. Our results demonstrated that 3,5-dicaffeoylquninic acid was the primary phenolic compounds in GJ that effectively reduced liver damage, providing a theoretical basis for the development of GJ as a potentially useful resource for hepatoprotective diet.

Platycodon grandiflorum saponins: Ionic liquid-ultrasound-assisted extraction, antioxidant, whitening, and antiaging activity.

This study explored the use of ionic liquid-ultrasound (ILU)-assisted extraction to enhance the extraction rate of Platycodon grandiflorum saponins (PGSs), and the content, extraction mechanism, antioxidant activity, whitening, and antiaging activity of PGSs prepared using ILU, ultrasound-water, thermal reflux-ethanol, and cellulase hydrolysis were compared. The ILU method particularly disrupted the cell wall, improved PGS extraction efficiency, and yielded a high total saponin content of 1.45 ± 0.02 mg/g. Five monomeric saponins were identified, with platycodin D being the most abundant at 1.357 mg/g. PGSs displayed excellent in vitro antioxidant activity and exhibited inhibitory effects on tyrosinase, elastase, and hyaluronidase. The results suggest that PGSs may have broad antioxidant, skin-whitening, and antiaging potential to a large extent. Overall, this study provided valuable insights into the extraction, identification, and bioactivities of PGSs, which could serve as a reference for future development and application of these compounds in the functional foods industry.

C-to-G editing generates double-strand breaks causing deletion, transversion and translocation.

Base editors (BEs) introduce base substitutions without double-strand DNA cleavage. Besides precise substitutions, BEs generate low-frequency 'stochastic' byproducts through unclear mechanisms. Here, we performed in-depth outcome profiling and genetic dissection, revealing that C-to-G BEs (CGBEs) generate substantial amounts of intermediate double-strand breaks (DSBs), which are at the centre of several byproducts. Imperfect DSB end-joining leads to small deletions via end-resection, templated insertions or aberrant transversions during end fill-in. Chromosomal translocations were detected between the editing target and off-targets of Cas9/deaminase origin. Genetic screenings of DNA repair factors disclosed a central role of abasic site processing in DSB formation. Shielding of abasic sites by the suicide enzyme HMCES reduced CGBE-initiated DSBs, providing an effective way to minimize DSB-triggered events without affecting substitutions. This work demonstrates that CGBEs can initiate deleterious intermediate DSBs and therefore require careful consideration for therapeutic applications, and that HMCES-aided CGBEs hold promise as safer tools.

Effect of grape seed proanthocyanidin on the structural and physicochemical properties of bread during bread fermentation stage.

Bread was prepared using wheat flour with grape seed proanthocyanidin (GSP) (0.4%). GSP improved the textural properties of bread including hardness, cohesiviness, gumminess and chewiness. At the last stage of fermentation, GSP reinforced the gluten microstructure with increased the disulfide bonds and hydrophobic interaction and α-helix in the secondary structures. Moreover, GSP addition could increase the total phenolics and antioxidative acitivity of the bread significantly. In addition, the degree of fermentation had a strong influence on the dough forces, and the reasonable control of bread fermentation time was beneficial to improve the bread quality, which provided a reference for the bread processing industry.

Insight into the phenolics and antioxidant activity of Indian jujube (Ziziphus mauritiana Lamk) peel and pulp subjected to the simulated digestion.

To evaluate the release and activity of Indian jujube phenolics in vivo, its peel and pulp were subjected to simulated digestions. The phenolics content and antioxidant activity of the digested samples were determined. The results showed that the total phenolics/flavonoids in the peel were respectively 4.63 and 4.48 times higher than that in the pulp. The release of phenolics and flavonoids respectively increased by 79.75% and 39.98% in the peel and 86.34% and 23.54% in the pulp after the intestinal digestion. The correlation between the total phenolics/flavonoids and antioxidant activity was higher in the peel (r > 0.858, p < 0.01) than that in the pulp. The phenolics profiles of the peel were almost the same after the digestion, and four phenolics including naringenin tri-glycoside, quercetin-3-O-[(2-hexosyl)-6-rhamnosyl] -hexoside, quercetin-3-O-pentosylhexoside and quercetin-3-O-(2-pentosyl -rhamnoside)-4'-O-rhamnoside were found to be the main flavonoids of Indian jujube peel, and they showed high recovery (>89.88%) during the digestion, implying that these phenolics may play a vital role in the function of Indian jujubes.

Profile and activity of phenolic antioxidants in chrysanthemum (Huangshan Gongju) as affected by simulated digestions.

The phenolics are the main bioactive substances of Huangshan Gongju, a famous chrysanthemum of China, but their digestive characteristics are still unknown. To explore the digestive properties of Huangshan Gongju phenolics, the flower was extracted and subjected to simulated digestions, and their phenolic profile and activity were analyzed. The results indicated that the total phenolics content and antioxidant activity of the extract varied with the simulated digestion steps, and they generally decreased in the oral and small intestine digestions but increased in the gastric digestion, and high correlations were detected between the total phenolics content and antioxidant activity (0.873 < r < 0.979, p < .01). The change of phenolic profile during the simulated digestions was similar to that of total phenolics content, and six individual phenolics were identified and quantified, and three of them, including chlorogenic acid, apigenin-7-O-rutinoside, and apigenin-7-O-6″-acetylglucoside showed higher recovery (>64.29%), implying they may be the main functional phenolics of Huangshan Gongju. PRACTICAL APPLICATIONS: This study proved that most phenolics in Huangshan Gongju were relatively stable during digestion. The finding may guarantee the application of Huangshan Gongju in the field of functional foods.

Evaluation of digestibility differences for apple polyphenolics using in vitro elderly and adult digestion models.

We evaluated the in vitro digestibility of apple polyphenols mimicking elderly and adult digestion models (dynamic and static systems). The digestibility of total apple polyphenols in small intestine was much higher in the adult dynamic system (62 µg/100 g fresh apple) compared to the static system (20 µg/100 g fresh apple) and elderly dynamic digestion conditions (33 µg/100 g fresh apple). Elderly in vitro static digestion showed better antioxidant activity than the adult system (OH and ABTS+ methods). Thus, the in vitro dynamic digestion system can more truly reflect the digestion of apple polyphenols than static digestion system. Moreover, elderly digestion conditions negatively influenced the digestibility of apple polyphenols including chlorogenic acid, epicatechin, phlorizin, rutin, phloretin, hyperoside, proanthocyanidin B2, and quercetin. Hence, appropriate selection of in vitro digestion models for elderly is a prerequisite to exploring the digestibility of phytochemicals for the development of functional food products for elderly.

Exploration of walnut components and their association with health effects.

Traditionally, walnuts have occupied an imperative position in the functional food market with consistently recognized nutritious and functional properties. In the past years, the lipid profile of walnuts has brought much scientific attention via linking a cascade of biological attributes and health-promoting effects. Over time, researchers have focused on diversified composition (polyphenols and vitamins) of different parts of walnut (flower, pellicle, and kernel) and emphasized their physiological significance. Consequently, a plethora of reports has emerged on the potential role of walnut consumption against a series of diseases including cancer, gut dysbiosis, cardiovascular, and neurodegenerative diseases. Therefore, we accumulated the updated data on composition and classification, extraction methods, and utilization of different parts of walnuts as well as associated beneficial effects under in vivo and clinical studies. Altogether, this review summarized the ameliorative effects of a walnut-enriched diet in chronic diseases which can be designated to the synergistic or individual effects of walnut components mainly through anti-oxidative and anti-inflammatory role.

Global detection of DNA repair outcomes induced by CRISPR-Cas9.

CRISPR-Cas9 generates double-stranded DNA breaks (DSBs) to activate cellular DNA repair pathways for genome editing. The repair of DSBs leads to small insertions or deletions (indels) and other complex byproducts, including large deletions and chromosomal translocations. Indels are well understood to disrupt target genes, while the other deleterious byproducts remain elusive. We developed a new in silico analysis pipeline for the previously described primer-extension-mediated sequencing assay to comprehensively characterize CRISPR-Cas9-induced DSB repair outcomes in human or mouse cells. We identified tremendous deleterious DSB repair byproducts of CRISPR-Cas9 editing, including large deletions, vector integrations, and chromosomal translocations. We further elucidated the important roles of microhomology, chromosomal interaction, recurrent DSBs, and DSB repair pathways in the generation of these byproducts. Our findings provide an extra dimension for genome editing safety besides off-targets. And caution should be exercised to avoid not only off-target damages but also deleterious DSB repair byproducts during genome editing.

Repair of programmed DNA lesions in antibody class switch recombination: common and unique features.

The adaptive immune system can diversify the antigen receptors to eliminate various pathogens through programmed DNA lesions at antigen receptor genes. In immune diversification, general DNA repair machineries are applied to transform the programmed DNA lesions into gene mutation or recombination events with common and unique features. Here we focus on antibody class switch recombination (CSR), and review the initiation of base damages, the conversion of damaged base to DNA double-strand break, and the ligation of broken ends. With an emphasis on the unique features in CSR, we discuss recent advances in the understanding of DNA repair/replication coordination, and ERCC6L2-mediated deletional recombination. We further elaborate the application of CSR in end-joining, resection and translesion synthesis assays. In the time of the COVID-19 pandemic, we hope it help to understand the generation of therapeutic antibodies.

Effect of grape seed power on the structural and physicochemical properties of wheat gluten in noodle preparation system.

Noodles were prepared using wheat flour supplemented with 1%, 3%, and 5% grape seed power (GSP). The farinograph properties of wheat flour, the textural properties of the dough, and thermal properties of the gluten were determined. The microstructure was analyzed by scanning electron and atomic force microscopy, and the effects of the addition of GSP on the physicochemical and structural properties (free sulfhydryl content, surface hydrophobic region, and secondary structure) of wheat gluten protein were analyzed. 1% GSP promoted the aggregation of gluten proteins by promoting hydrophobic interactions and hydrogen bonding, thus enhanced the noodle quality. Whereas, 3% and 5% GSP addition disrupted the disulfide bonds between gluten protein molecules and formed macromolecular aggregates linked to gluten proteins through non-covalent bonds and hydrophobic interactions, which prevented the formation of the gluten protein reticulation structure. Our study emphasized the interaction between wheat proteins and GSP in noodle making dough.

Solvent effect on phenolics and antioxidant activity of Huangshan Gongju (Dendranthema morifolium (Ramat) Tzvel. cv. Gongju) extract.

Huangshan Gongju was extracted with organic solvents (ethanol, methanol and acetone) of different concentrations (0-90%), and the extracts' phenolic content and antioxidant activity, as well as the correlations between them were examined. With the increasing concentration of organic solvent, the total phenolic compound (TPC) increased continuously and met its maximum at 70% acetone, whereas the total flavonoid compound (TFC) and most individual phenolics met their maximums at 70% ethanol. Similar changes occurred to the antioxidant activity, including DPPH and ABTS scavenging activities, and their maximums were respectively found at 50% acetone and 70% ethanol. The antioxidant activity correlated strongly with TPC/TFC (r > 0.954, p < 0.01) and individual phenolics (r > 0.886, p < 0.05), and the strongest correlations between them were mainly given by luteolin-7-O-glucoside (r > 0.975, p < 0.001). These results suggested that high content organic solvent (50-70%) was beneficial to obtain Huangshan Gongju extracts of higher phenolic content and antioxidant activity, and 70% ethanol may be the promising solvent. Besides, phenolics were found to be the main antioxidants of Huangshan Gongju extracts, and flavonoids especially luteolin-7-O-glucoside may play more important roles in the antioxidant activity.

A new isoflavone glycoside from flowers of Pueraria Montana var. lobata (Willd.) Sanjappa & Pradeep.

A new isoflavone glycoside, named 3'-hydroxytectorigenin-7-O-ß-D-xylosyl-(1→6)-ß-D-glucopyranoside (1) was isolated from the flowers of Pueraria montana var. lobata (Willd.) Sanjappa & Pradeep. The structure of compound 1 was characterised by HR-ESI-MS and NMR spectroscopic methods. In radical scavenging activity test using 2, 2-diphenyl-1-picrylhydrazyl (DPPH), compound 1 showed moderate activity with IC50 value of 42 ± 4.2 µg/mL.

Genome integrity and neurogenesis of postnatal hippocampal neural stem/progenitor cells require a unique regulator Filia.

Endogenous DNA double-strand breaks (DSBs) formation and repair in neural stem/progenitor cells (NSPCs) play fundamental roles in neurogenesis and neurodevelopmental disorders. NSPCs exhibit heterogeneity in terms of lineage fates and neurogenesis activity. Whether NSPCs also have heterogeneous regulations on DSB formation and repair to accommodate region-specific neurogenesis has not been explored. Here, we identified a regional regulator Filia, which is predominantly expressed in mouse hippocampal NSPCs after birth and regulates DNA DSB formation and repair. On one hand, Filia protects stalling replication forks and prevents the replication stress-associated DNA DSB formation. On the other hand, Filia facilitates the homologous recombination-mediated DNA DSB repair. Consequently, Filia-/- mice had impaired hippocampal NSPC proliferation and neurogenesis and were deficient in learning, memory, and mood regulations. Thus, our study provided the first proof of concept demonstrating the region-specific regulations of DSB formation and repair in subtypes of NSPCs.

REV7 is required for processing AID initiated DNA lesions in activated B cells.

Activation-induced cytidine deaminase (AID) initiates both antibody class switch recombination (CSR) and somatic hypermutation (SHM) in antibody diversification. DNA double-strand break response (DSBR) factors promote rearrangement in CSR, while translesion synthesis (TLS) polymerases generate mutations in SHM. REV7, a component of TLS polymerase zeta, is also a downstream effector of 53BP1-RIF1 DSBR pathway. Here, we study the multi-functions of REV7 and find that REV7 is required for the B cell survival upon AID-deamination, which is independent of its roles in DSBR, G2/M transition or REV1-mediated TLS. The cell death in REV7-deficient activated B cells can be fully rescued by AID-deficiency in vivo. We further identify that REV7-depedent TLS across UNG-processed apurinic/apyrimidinic sites is required for cell survival upon AID/APOBEC deamination. This study dissects the multiple roles of Rev7 in antibody diversification, and discovers that TLS is not only required for sequence diversification but also B cell survival upon AID-initiated lesions.

Effects of water-immersion cooling temperatures on the moisture retention of sodium-reduced pork sausages.

To improve the industrial yield of sodium-reduced meat products, we present a feasible method by adjusting water-immersion cooling temperatures to decrease the water loss of pork sausage during processing. The present results showed that the moisture retention capacity of sodium-reduced pork sausages (SRPS) cooled by the temperatures of 15-20 °C was larger than that of 0-10 °C. The higher cooling temperatures, especially at 20 °C, could change the movement and population of proton of inner water, decrease syneresis and facilitate the formation of homogenous cross-linked network, thus increasing the moisture retention of SRPS. Results of this work indicated that the control of cooling temperature of sodium-reduced sausages after cooking could serve as a feasible approach for improving the economic benefits and quality characteristics of the final products.

ERCC6L2 promotes DNA orientation-specific recombination in mammalian cells.

Programmed DNA recombination in mammalian cells occurs predominantly in a directional manner. While random DNA breaks are typically repaired both by deletion and by inversion at approximately equal proportions, V(D)J and class switch recombination (CSR) of immunoglobulin heavy chain gene overwhelmingly delete intervening sequences to yield productive rearrangement. What factors channel chromatin breaks to deletional CSR in lymphocytes is unknown. Integrating CRISPR knockout and chemical perturbation screening we here identify the Snf2-family helicase-like ERCC6L2 as one such factor. We show that ERCC6L2 promotes double-strand break end-joining and facilitates optimal CSR in mice. At the cellular levels, ERCC6L2 rapidly engages in DNA repair through its C-terminal domains. Mechanistically, ERCC6L2 interacts with other end-joining factors and plays a functionally redundant role with the XLF end-joining factor in V(D)J recombination. Strikingly, ERCC6L2 controls orientation-specific joining of broken ends during CSR, which relies on its helicase activity. Thus, ERCC6L2 facilitates programmed recombination through directional repair of distant breaks.

Comparison of phenolic compounds extracted from Diaphragma juglandis fructus, walnut pellicle, and flowers of Juglans regia using methanol, ultrasonic wave, and enzyme assisted-extraction.

The phenols in Diaphragma juglandis fructus (DJF), walnut pellicle (WP), and flowers of Juglans regia (FJR) from walnut were extracted using three methods (methanolic condensation reflux extraction, ultrasonic wave extraction, and enzyme assisted-extraction), and phenolics and antioxidant capacities of different extractions were compared. Overall, 50 phenolics were identified by HPLC-MS/MS with 41 compounds in DJF, 32 in WP, and 29 in FJR. It was observed that tannins in WP was higher than those in DJF and FJR. As for PCA, more than 70% of the variance was explained with the obvious comparison between the phenolic constituents. The phenolics in walnut contributed to remarkable antioxidant effect, with the highest effect observed in WP. This study presents the analysis and comparison of the phenols can be further extended for the development of functional walnut instant foods.

Phenolics and antioxidant activity of bamboo leaves soup as affected by in vitro digestion.

Bamboo leaves soups were subjected to in vitro digestion (including separated oral, gastric and small intestinal digestions, and complete digestion containing above three stages), and their phenolics and antioxidant activities were determined. Compared to control groups, total phenolic content (TPC) in treated groups (including undigested and digested groups) increased at gastric digestion stage but decreased at other digestion stages, and the decrease in small intestinal digestion stage (19.97%) was nearly the same with that in complete digestion stage (19.39%). The antioxidant activity in digested groups almost changed accordingly to their TPC but with no significant difference (p > 0.05) as compared with undigested groups; similar results were found in four main individual phenolics including cryptochlorogenic acid, chlorogenic acid, neochlorogenic acid and isoorientin, and their contents were negatively correlated to the pH value of digestion buffers (-0.68 < r < -0.80, p < 0.01). These results indicated that the change of phenolic content and antioxidant activity in digested bamboo leaves soups mainly resulted from the pH of digestion buffers rather than digestive enzymes. In addition, the decrease of phenolics may mainly occur at small intestinal digestion stage where the pH value is the highest in the digestive system.

Comparative study of chemical compositions and antioxidant activities of Zhizi fruit extracts from different regions.

The fruits of Gardenia jasminoides Ellis are folk medicines in China and their major components are geniposide and water soluble pigment crocins. This study compared the chemical profiles and free radical scavenging activities of two Zhizi species from five provinces of China, including Jiangxi, Anhui, Hunan, Sichuan and Henan. The results showed that Jiangxi Zhizi contained higher levels of volatiles (71.84%), crocins (20.38 mg/g), geniposide (31.36 mg/g) and flavonoids (84.42 µg quercetin/mg) than four other Zhizi fruits; whereas Hunan Zhizi provided higher total phenolics (33.81 µg catechin/mg) and ABTS/DPPH radical scavenging activities. These findings implied that Jiangxi Zhizi would be suitable for extraction of gardenia yellow and geniposide, as well as preparation of essential oil. This information may provide valuable guidance for application of Zhizi fruits to biomedicine industry in China.