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This study examines the impact of blanching (heating at 85 °C for 60 s), high-pressure processing (HPP) (600 MPa, 3 min, 20 °C), and a combination of both blanching and HPP on the microbiological and chemical qualities, colour, and antioxidant properties of carrot juice stored at 4 °C for 15 days. In terms of microbiological quality, the total plate count (TPC), coliform bacteria, and Salmonella spp. rose rapidly in the control group (untreated) as the storage time increased. However, for the blanching group, these values climbed more gradually, surpassing the microbiological limits for juice beverages (TPC < 4 log CFU/mL, Coliform < 10 MPN/mL, and Salmonella spp. negative) on the 9 days of storage. In contrast, TPC, coliforms, and Salmonella spp. were undetectable in the HPP and blanching/HPP samples throughout the storage period. Additionally, as storage time lengthened, the pH, total soluble solids, and Hunter colour values ( L , a , b ) diminished in the control and blanching groups, whilst titratable acidity and browning degree intensified. However, the HPP and blanching/HPP noticeably delayed these decreases or increases. Moreover, although the total phenolic content and DPPH radical scavenging ability in the HPP samples remained relatively stable during storage and were lower compared to other groups, the β-carotene content was higher at the end of the storage period. In summary, HPP can effectively deactivate microorganisms in carrot juice, irrespective of whether blanching is applied, and can impede reductions in pH, increases in acidity, and colour changes, ultimately extending the juice's shelf life.

This study aimed to assess the use of the high-hydrostatic-pressure (HHP) method (200-600 MPa, 5 min) for bleaching mustard pickle products as an alternative to the conventional method of sulfite addition. The aerobic plate count (APC) and lactic acid bacteria count (LAB) of the samples decreased with the increase in pressure, and the yeast count decreased to no detectable levels. Next, compared with the control group (no high-pressure treatment) the L * (lightness), W (whiteness), Δ E (color difference), and texture (hardness and chewiness) of the HHP-processed samples, which increased significantly with increasing pressure, while the a* (redness) and b* (yellowness) values decreased slightly. This indicates that HHP processing gave the mustard pickle a harder texture and a brighter white color and appearance. Furthermore, when the mustard pickle was treated with HHP 400 and 600 MPa for 5 min and stored at 25 °C for 60 days, it was found that the APC and LAB counts in the HHP-processed group recovered rapidly and did not differ from those in the control group (the non-HHP treated group) but significantly delayed the growth of yeast, the increase in pH value, and total volatile basic nitrogen (TVBN). The high-throughput sequencing (HTS) analysis revealed that the predominant bacterial genera in the non-HHP-treated mustard pickle were Lactiplantibacillus (74%), Lactilactobacillus (12%), and Levilactobacillus (6%); after 60 days of storage, Companilactobacillus (80%) became dominant. However, after 60 days of storage, Lactiplantibacillus (92%) became dominant in the samples processed at 400 MPa, while Levilactobacillus (52%), Pediococcus (17%), and Lactiplantibacillus (17%) became dominant in the samples processed at 600 MPa. This indicated that the HHP treatment changed the lactic acid bacterial flora of the mustard pickle during the storage period. Overall, it is recommended to treat the mustard pickle with HHP above 400 MPa for 5 min to improve its texture and color and delay the deterioration of quality during storage. Therefore, HHP technology has the potential to be developed as a treatment technique to replace the addition of sulfite.

Particulate matter (PM) is one of the reasons that exacerbate skin diseases. Impaired barrier function is a common symptom in skin diseases, including atopic dermatitis, eczema and psoriasis. Herbal extracts rich in antioxidants are thought to provide excellent pharmacological activities; however, the anti-pollution activity of Artocarpus altilis extract (AAM) has not been investigated yet. The present study demonstrated that 5 μg/mL of AAM was considered to be a safe dose for further experiments without cytotoxicity. Next, we evaluated the anti-pollution activity of AAM through the PM-induced keratinocytes damage cell model. The results showed that AAM could reduce PM-induced overproduction of intracellular ROS and the final product of lipid peroxidation, 4-hydroxynonenal (4HNE). In addition, AAM not only reduced the inflammatory protein expressions, including tumor necrosis factor α (TNFα), TNF receptor 1 (TNFR1) and cyclooxygenase-2 (COX-2), but also balanced the aging protein ratio of matrix metalloproteinase (MMPs) and tissue inhibitors of metalloproteases (TIMPs) through downregulating the phosphorylation of mitogen-activated protein kinase (MAPK) signaling. For skin barrier protection, AAM could repair PM-induced barrier function proteins damage, including filaggrin, loricrin and aquaporin 3 for providing anti-aging bioactivity. In conclusion, AAM has the potential to be developed as an anti-pollution active ingredient for topical skin products to prevent skin oxidation, inflammation and aging, and restore the skin barrier function.

Ultraviolet B (UVB) is one of the most important environmental factors that cause extrinsic aging through increasing intracellular reactive oxygen species (ROS) production in the skin. Due to its protective roles against oxidative stress, nuclear factor erythroid-2-related factor (NRF2) has been traditionally considered as a target for skin aging prevention. Here, we identified the extract of Prinsepiae Nux, a top-grade drug listed in Shen Nong Ben Cao Jing, as a potent NRF2 activator by high-throughput screening. A bioassay-guided fractionation experiment revealed that NRF2-activating components were concentrated in the 90% methanol (MP) fraction. MP fraction significantly increased the expression of NRF2 and HO-1 protein and upregulated HO-1 and NQO1 mRNA expression in HaCaT cells. Moreover, MP fraction pre-treatment dramatically reversed UVB-induced depletion of NRF2 and HO-1, accumulation of intracellular ROS, NF-κB activation, and the upregulation of pro-inflammatory genes. Finally, the qualitative analysis using UPLC-tandem mass spectroscopy revealed the most abundant ion peak in MP fraction was identified as α-linolenic acid, which was further proved to activate NRF2 signaling. Altogether, the molecular evidence suggested that MP fraction has the potential to be an excellent source for the discovery of natural medicine to treat/prevent UVB-induced skin damage.

Recently, a global market for anti-aging skin care using botanicals has been noticeably developing. Morin, 3,5,7,2',4'-pentahydroxyflavone, is a polyphenol with many pharmacological properties including antioxidant, anti-inflammation and photoprotection. However, poor aqueous solubility of morin restricts its application in pharmaceuticals. The present study aimed to encapsulate morin into liposomal vesicles to improve its water solubility and skin penetration, and further investigated its ROS inhibition and anti-aging activity in HaCaT keratinocytes induced by particulate matters (PMs). Our data presented that morin was a strong DPPH • radical scavenger. Morin displayed a remarkable ROS inhibitory ability and protected keratinocytes against PMs by downregulating matrix metalloproteinase-1 (MMP-1) expression via suppressing p-ERK and p-p38 in the MAPK pathway. Moreover, water solubility of liposomal morin (LM) prepared by the thin film hydration method was significantly better than free form of morin due to particle size reduction of LM. Our results also demonstrated that deformable liposomal vesicles were achieved for increasing dermal absorption. Additionally, LM (morin:lecinolws-50:tween-80:PF-68, 1:2.5:2.5:5) was able to effectively reduce generation of ROS, inactivate p-ERK, p-p38 and MMP-1 in HaCaT cells exposed to PM. In conclusion, our findings suggested that LM would be a bright candidate for various topical anti-aging and anti-pollution products.

Particulate matter (PM) is the main indicator of air pollutants, and it may increase the level of reactive oxygen species (ROS) in keratinocytes, leading to skin inflammation, aging, and decreased moisturizing ability. Pterostilbene (PTS) is a dimethylated analog of resveratrol that has antioxidant effects. However, the molecular mechanisms of PTS in preventing PM-induced keratinocyte inflammation and aging have not been investigated yet. Therefore, we used PM-induced human keratinocytes to investigate the protective mechanisms of PTS. The results showed that 20 μM PTS had no toxicity to HaCaT keratinocytes and significantly reduced PM-induced intracellular ROS production. In addition, nuclear translocation of the aryl hydrocarbon receptor (AHR) was inhibited by PTS, leading to reduced expression of its downstream CYP1A1. PTS further inhibited PM-induced MAPKs, inflammation (COX-2), and aging (MMP-9) protein cascades, and rescued moisturizing (AQP-3) protein expression. We analyzed the PTS content in cells at different time points and compared the concentration required for PTS to inhibit the target proteins. Finally, we used the skin penetration assay to show that the PTS essence mainly exists in the epidermal layer and did not enter the system circulation. In conclusion, PTS could protect HaCaT keratinocytes from PM-induced damage and has the potential to become a cosmetic ingredient.

Airborne particulate matter (PM) is one of the indicators of air pollution, and it is also the main factor causing oxidative stress in the skin. Oleanolic acid (OA), a natural terpenoid compound, effectively inhibited PM-induced skin aging; however, OA has poor water solubility and skin absorption, which limit its application in medicines and cosmetics. The aim of this study was to prepare oleanolic acid nanofibers (OAnf) and evaluate the effects of OA and OAnf in PM-treated keratinocytes. The results showed that OA dissolved in dissolved in dimethyl sulfoxide (DMSO) attenuated PM-induced reactive oxygen species overproduction, stress-activated protein kinase/Jun-amino-terminal kinase (SAPK/JNK) activation, and the expressions of inflammatory and skin-aging-related proteins. In addition, the nanofiber process of OA effectively improved the water solubility of OA more than 99,000-fold through changing its physicochemical properties, including a surface area increase, particle size reduction, amorphous transformation, and hydrogen bonding formation with excipients. The skin penetration ability of OAnf was consistently over 10-fold higher than that of OA. Moreover, when dissolved in PBS, OAnf displayed superior antioxidant, anti-inflammatory, and anti-skin aging activities in PM-treated keratinocytes than OA. In conclusion, our findings suggest that OAnf could be a topical antioxidant formulation to attenuate skin problems caused by PM.

Transcription factor nuclear factor erythroid 2-related factor 2 (NRF2) plays a crucial role in regulating the expression of genes participating in cellular defense mechanisms against oxidative or xenobiotic insults. However, there is increasing evidence showing that hyperactivation of NRF2 is associated with chemoresistance in several cancers, including hepatocellular carcinoma (HCC), thus making NRF2 an attractive target for cancer therapy. Another important issue in cancer medication is the adverse effects of these substances on normal cells. Here, we attempted to identify a dual-selective NRF2 regulator that exerts opposite effects on NRF2-hyperactivated HCC cells and normal keratinocytes. An antioxidant response element driven luciferase reporter assay was established in Huh7 and HaCaT cells as high-throughput screening platforms. Screening of 3,000 crude extracts from the Taiwanese Indigenous Plant Extract Library resulted in the identification of Beilschmiedia tsangii (BT) root extract as a dual-selective NRF2 regulator. Multiple compounds were found to contribute to the dual-selective effects of BT extract on NRF2 signaling in two cell lines. BT extract reduced NRF2 protein level and target gene expression levels in Huh7 cells but increased them in HaCaT cells. Furthermore, notable combinatory cytotoxic effects of BT extract and sorafenib on Huh7 cells were observed. On the contrary, sorafenib-induced inflammatory reactions in HaCaT cells were reduced by BT extract. In conclusion, our results suggest that the combination of a selective NRF2 activator and inhibitor could be a practical strategy for fine-tuning NRF2 activity for better cancer treatment and that plant extracts or partially purified fractions could be a promising source for the discovery of dual-selective NRF2 regulators.