Identification and quality evaluation of Lushan Yunwu tea from different geographical origins based on metabolomics.

The relationship between the chemical composition and quality of Lushan Yunwu tea (LYT) from different geographical origins is not clear. Sensory evaluation, metabolomics analyses combined with chemometrics were conducted on LYT from 8 different geographical origins, and altitude was identified as the main factor responsible for the differences among LYT. A total of 32 non-volatile and 27 volatile compounds were identified as marker metabolites to distinguish the origins of high altitudes from those of low altitudes. LYT samples from higher altitude areas contained more free amino acids, sugars, and organic acids, and less catechins, which may contribute to the reduction of bitterness and astringency and the enhancement of umami. The contents of geranylacetone, ethyl hexanoate, ethyl caprylate, 3-carene, d-cadinene, linalool, nerol, and nerolidol in high altitude areas were higher than those in low altitude areas, indicating that LYT from high altitude had strong floral and fruity aroma. The altitudes were positively correlated with pH value, total flavonoids, soluble protein, total free amino acids, and the antioxidant capacities of the LYT. This study provided a theoretical basis for the study of the effect of altitude on tea quality.

Fungal fermentation of Fuzhuan brick tea: A comprehensive evaluation of sensory properties using chemometrics, visible near-infrared spectroscopy, and electronic nose.

Fuzhuan brick tea (FBT) fungal fermentation is a key factor in achieving its unique dark color, aroma, and taste. Therefore, it is essential to develop a rapid and reliable method that could assess its quality during FBT fermentation process. This study focused on using electronic nose (e-nose) and spectroscopy combination with sensory evaluations and physicochemical measurements for building machine learning (ML) models of FBT. The results showed that the fused data achieved 100 % accuracy in classifying the FBT fermentation process. The SPA-MLR method was the best prediction model for FBT quality (R2 = 0.95, RMSEP = 0.07, RPD = 4.23), and the fermentation process was visualized. Where, it was effectively detecting the degree of fermentation relationship with the quality characteristics. In conclusion, the current study's novelty comes from the established real-time method that could sensitively detect the unique post-fermentation quality components based on the integration of spectral, and e-nose and ML approaches.

Green tea extract suppresses airway inflammation via oxidative stress-driven MAPKs/MMP-9 signaling in asthmatic mice and human airway epithelial cells.

Introduction: The anti-inflammatory effect of green tea extract (GTE) has been confirmed in asthmatic mice, however, the pharmacological mechanism is not fully elucidated. Methods: To investigate the therapeutic efficacy of GTE in asthma and identify specific pathways, murine model of allergic asthma was established by ovalbumin (OVA) sensitization and the challenge for 4 weeks, with oral treatment using GTE and dexamethasone (DEX). Inflammatory cell counts, cytokines, OVA-specific IgE, airway hyperreactivity, and antioxidant markers in the lung were evaluated. Also, pulmonary histopathological analysis and western blotting were performed. In vitro, we established the model by stimulating the human airway epithelial cell line NCI-H292 using lipopolysaccharide, and treating with GTE and mitogen-activated protein kinases (MAPKs) inhibitors. Results: The GTE100 and GTE400 groups showed a decrease in airway hyperresponsiveness and the number of inflammatory cells in the bronchoalveolar lavage fluid (BALF) compared to the OVA group. GTE treatment also reduced interleukin (IL)-13, IL-5, and IL-4 levels in the BALF, and OVA-specific immunoglobulin E levels in the serum compared to those in the OVA group. GTE treatment decreased OVA-induced mucus secretion and airway inflammation. In addition, GTE suppressed the oxidative stress, and phosphorylation of MAPKs, which generally occurs after exposure to OVA. GTE administration also reduced matrix metalloproteinase-9 activity and protein levels. Conclusion: GTE effectively inhibited asthmatic respiratory inflammation and mucus hyperproduction induced by OVA inhalation. These results suggest that GTE has the potential to be used for the treatment of asthma.

A new sustainable filtration membrane extraction method based on nanoconfined liquid phase extraction: Determination of organochlorinated pesticides and pyrethroids in tea samples.

BACKGROUND: Pesticides are used in agricultural production for prevent and control crop diseases and pests, but it is easy to cause excessive pesticides residues in agricultural products, polluting the environment and endangering human health. Due to their unmatched and sustainable capabilities, nanoextraction procedures are becoming every day more important in Analytical Chemistry. In particular, nanoconfined liquid phase extraction has shown extraction capabilities toward polar, medium polar, and/or nonpolar substances, which can be easily modulated depending on the nanoconfined solvent used. Furthermore, this "green" technique showed excellent characteristics in terms of recoveries, extraction time (≤1 min), reliability, and versatility. (97) RESULTS: In this work, the advantages of this technique have been coupled with those of filtration membrane extraction, making use of carbon nanofibers (CnFs) growth on carbon microspheres (CµS). This substrate has been deposited on a filter, which combined with gas chromatographic mass spectrometry (GC-MS) analysis successfully employed for the nanoextraction of 30 pesticides (18 organochlorine and 12 pyrethroids) in tea samples. Under the optimized extraction conditions, the linear range with standard solutions was from 1 to 1000 ng mL-1 (R2 ≥ 0.99), the limit of detections in tea samples were in the range 0.56-17.98 µg kg-1. The accuracy of the developed method was evaluated by measuring the extraction recovery of the spiked tea samples, and recoveries between 74.41 % and 115.46 %. (119) SIGNIFICANCE: Considering the versatility of nanoconfined liquid phase extraction and the functionality of the filtration membrane extraction procedure, this new extraction method can be considered a powerful candidate for automatized high-throughput analyses of real samples. (34).

Simultaneous determination of six catechins and caffeine in tea and wine using salting-out assisted liquid-liquid extraction and high-performance liquid chromatography with ultraviolet detection.

Catechins, renowned for their antioxidant properties and health benefits, are commonly present in beverages, particularly tea and wine. An efficient and cost-effective salting-out assisted liquid-liquid extraction (SALLE) method has been developed and validated for the simultaneous determination of six catechins and caffeine in tea and wine samples using high-performance liquid chromatography-ultraviolet (HPLC-UV). This method demonstrates outstanding performance: linearity (1-120 µg/mL, r2 > 0.999), accuracy (96.5%-103.4% recovery), and precision (≤14.7% relative standard deviation), meeting validation requirements set by the US Food and Drug Administration. The reduced sample size (0.1 g) minimizes matrix interferences and costs without compromising sensitivity. All analytes were detected in Camellia sinensis teas, with green tea displaying the highest total catechin content (47.5-100.1 mg/mL), followed by white and black teas. Analysis of wine samples reveals the presence of catechin in all red and white wines, and epigallocatechin gallate in all red wine samples, highlighting the impact of winemaking processes on catechin content. The SALLE-HPLC-UV approach represents a green alternative by eliminating organic waste, surpassing conventional dilution methods in specificity and sensitivity for catechin determination. AGREEprep assessment emphasizes the strengths of the SALLE procedure, including material reusability, throughput efficiency, minimal sample requirements, low energy consumption, and the absence of organic waste generation.

Changes in amino acids, catechins and alkaloids during the storage of oolong tea and their relationship with antibacterial effect.

The storage process has a significant impact on tea quality. Few is known about effect of storage on quality of oolong tea. This study aimed to assess the effect of different storage times on the key chemical components of oolong tea by measuring changes in catechin, free amino acid, and alkaloid content. Variation in the main substances was determined by principal component analysis and heat map analysis. The results revealed notable effects of the storage process on the levels of theanine, epigallocatechin gallate (EGCG), and glutamine. These findings suggest that these compounds could serve as indicators for monitoring changes in oolong tea quality during storage. Additionally, the study observed an increase in the antibacterial ability of tea over time. Correlation analysis indicated that the antibacterial ability against Micrococcus tetragenus and Escherichia coli was influenced by metabolites such as aspartic acid, threonine, serine, gamma-aminobutyric acid, ornithine, alanine, arginine, and EGCG. Overall, this study presents an approach for identifying key metabolites to monitor tea quality effectively with relatively limited data.

Investigating the Impact of Tea Consumption on Cognitive Function and Exploring Tea-Genetic Interactions in Older Adults Aged 65-105 Years: Findings from the 2002-2018 CLHLS Data.

BACKGROUND: As the global population ages, cognitive impairment (CI) becomes more prevalent. Tea has been one of the most popular drinks in the world. Several studies have demonstrated that tea consumption has an impact on cognitive function. OBJECTIVE: This study aims to examine the association between tea consumption and cognitive function and explore the potential effect of genetics on the relationship between tea consumption and CI risk in older adults. DESIGN: This is a prospective longitudinal study using data from the Chinese Longitudinal Healthy Longevity Survey (CLHLS). SETTING: Six waves of data from CLHLS containing 76,270 subjects were analyzed. Generalized estimation equations (GEE) with a logit link function were adopted to estimate the effect of tea consumption on CI risk from a cross-sectional and longitudinal perspective. PARTICIPANTS: A population-based cohort of adults aged 65-105 years. MEASUREMENTS: The frequency and type of tea consumption were obtained by questionnaires. CI was measured based on MMSE. Polygenic risk was measured using the polygenic score approach described by the International Schizophrenia. RESULTS: The results showed that drinking green tea had a better protective effect on cognitive function than other types of tea, the incidence of CI gradually decreased with the increase of tea consumption frequency, and men were more likely to benefit from tea consumption. Additionally, we also found a significant interaction between tea consumption and genetic risk, measured by polygenic risk score (PRS). CONCLUSIONS: Based on current research evidence, tea consumption, may be a simple and important measure for CI prevention.

Color Stability of Bioactive Restorative Material vs Nanohybrid Resin Composite: An In Vitro Study.

AIM: This study aimed to assess the color stability of bioactive restorative materials vs nanohybrid resin composites after 3 months of immersion in three frequently consumed beverages. MATERIALS AND METHODS: Thirty disk-shaped specimens of Giomer dental restorative material (Shofu, Japan) and nanohybrid resin composite (Tokuyama, Japan) were performed using a Teflon mold. Super-Snap system (Shofu, Japan) was utilized to finish and polish the specimens to be preserved for 24 hours in distilled water at 37°C. The samples had been divided into three subgroups (Coffee, tea, Pepsi) (n = 5). The initially displayed color measurements of the samples were performed using a spectrophotometer (VITA Easyshade® V). After 7 days, 30 days, and 90 days, color measurements were repeated, and the E of each sample was estimated. E of each sample was calculated. RESULTS: The Giomer group showed statistically significant higher E values than the nanohybrid resin composite where the p-value was ≤0.0001. Tea subgroup showed the highest statistically significant E values in both groups where the p-value was ≤ 0.0001. The highest statistically significant color change was recorded at 3 months. CONCLUSION: The color of bioactive restorative material is less stable if compared with nanohybrid resin composite. CLINICAL SIGNIFICANCE: As tea and coffee are popular beverages, particularly in Middle Eastern nations, dentists must advise patients about the color change of resin restorations. Patients are advised to brush their teeth immediately after consuming these beverages. How to cite this article: Saber EH, Abielhassan MH, Abed YA, et al. Color Stability of Bioactive Restorative Material vs Nanohybrid Resin Composite: An In Vitro Study. J Contemp Dent Pract 2024;25(3):221-225.

The effects of green tea tablets and metformin on ovulation and menstrual cycle regularity in women with polycystic ovary syndrome.

Polycystic ovary syndrome is the most common cause of oligo-ovulation and anovulation among women of reproductive age, contributing to infertility. This study aimed to compare the effects of green tea tablets and metformin on ovulation, menstrual cycle regularity, and antioxidant biomarkers in women with polycystic ovary syndrome (PCOS). In this clinical trial study, 94 women with PCOS were randomly assigned to three groups: green tea (n = 33), metformin (n = 29), and control (n = 32). Menstrual status and oxidative stress parameters, including total antioxidant capacity, thiol, and lipid peroxidation, were compared before and 3 months after the intervention among all three groups. Data analysis was conducted using SPSS software version 22 and employing the analysis of variance and paired t-tests. Following the intervention, the mean menstrual cycle duration in the green tea, metformin, and control groups was 32.22 ± 12.78, 48.72 ± 37.06, and 48.53 ± 31.04 days, respectively (P = 0.040). There was no statistically significant difference between the three groups in terms of biochemical, hormonal, and antioxidant indices before and after the intervention (P > 0.05). The intake of green tea tablets was associated with better outcomes in regulating the menstrual cycle in women with PCOS.

YOLOv8-RMDA: Lightweight YOLOv8 Network for Early Detection of Small Target Diseases in Tea.

In order to efficiently identify early tea diseases, an improved YOLOv8 lesion detection method is proposed to address the challenges posed by the complex background of tea diseases, difficulty in detecting small lesions, and low recognition rate of similar phenotypic symptoms. This method focuses on detecting tea leaf blight, tea white spot, tea sooty leaf disease, and tea ring spot as the research objects. This paper presents an enhancement to the YOLOv8 network framework by introducing the Receptive Field Concentration-Based Attention Module (RFCBAM) into the backbone network to replace C2f, thereby improving feature extraction capabilities. Additionally, a mixed pooling module (Mixed Pooling SPPF, MixSPPF) is proposed to enhance information blending between features at different levels. In the neck network, the RepGFPN module replaces the C2f module to further enhance feature extraction. The Dynamic Head module is embedded in the detection head part, applying multiple attention mechanisms to improve multi-scale spatial location and multi-task perception capabilities. The inner-IoU loss function is used to replace the original CIoU, improving learning ability for small lesion samples. Furthermore, the AKConv block replaces the traditional convolution Conv block to allow for the arbitrary sampling of targets of various sizes, reducing model parameters and enhancing disease detection. the experimental results using a self-built dataset demonstrate that the enhanced YOLOv8-RMDA exhibits superior detection capabilities in detecting small target disease areas, achieving an average accuracy of 93.04% in identifying early tea lesions. When compared to Faster R-CNN, MobileNetV2, and SSD, the average precision rates of YOLOv5, YOLOv7, and YOLOv8 have shown improvements of 20.41%, 17.92%, 12.18%, 12.18%, 10.85%, 7.32%, and 5.97%, respectively. Additionally, the recall rate (R) has increased by 15.25% compared to the lowest-performing Faster R-CNN model and by 8.15% compared to the top-performing YOLOv8 model. With an FPS of 132, YOLOv8-RMDA meets the requirements for real-time detection, enabling the swift and accurate identification of early tea diseases. This advancement presents a valuable approach for enhancing the ecological tea industry in Yunnan, ensuring its healthy development.

A V-shaped bis-coumarin based fluorescence probe for F- detection in tea infusions and potable water and bioimaging applications in living systems.

Fluorine (F) is a pivotal element in the formation of human dental and skeletal tissues, and the consumption of water and tea constitutes a significant source of fluoride intake. However, prolonged ingestion of water and tea with excessive fluoride content can lead to fluorosis, which poses a serious health hazard. In this manuscript, a novel turn-on fluorescent probe DCF synthesized by bis-coumarin and tert-butyldiphenylsilane (TBDPS) was introduced for detecting F- in potable water and tea infusions. By leveraging the unique chemical affinity between fluoride and silicon, F- triggers the silicon-oxygen bond cleavage in DCF, culminating in a conspicuous emission of yellow fluorescence. Validated through a succession of optical tests, this probe exhibits remarkable advantages in terms of superior selectivity, a low detection limit, a large Stokes shift, and robust interference resistance when detecting inorganic fluoride. Moreover, it can serve as portable test strips for on-site real-time identification and quantitative analysis of F-. Furthermore, the application of DCF for in-situ monitoring and imaging of F- in zebrafish and soybean root tissues proved its significant value for F- detection in both animal and plant systems. This probe potentially functions as an efficient instrument for delving into the toxic mechanisms of fluoride in physiological processes.

Intercropping fruit trees in tea plantation improves soil properties and the formation of tea quality components.

Intercropping has been recommended as a beneficial cropping practice for improving soil characteristic and tea quality. However, there is limited research on the effects of intercropping fruit trees on soil chemical properties, soil aggregate structure, and tea quality components. In this study, intercropping fruit trees, specifically loquats and citrus, had a significant impact on the total available nutrients, AMN, and AP in soil. During spring and autumn seasons, the soil large-macroaggregates (>2 mm) proportion increased by 5.93% and 19.03%, as well as 29.23% and 19.14%, respectively, when intercropping loquats and citrus. Similarly, intercropping waxberry resulted in a highest small-macroaggregates (0.25 mm-2 mm) proportion at 54.89% and 77.32%. Soil aggregate stability parameters of the R0.25, MWD, and GMD were generally considered better soil aggregate stability indicators, and significantly improved in intercropping systems. Intercropping waxberry with higher values for those aggregate stability parameters and lower D values, showed a better soil aggregate distribution, while intercropping loquats and citrus at higher levels of AMN and AP in different soil aggregate sizes. As the soil aggregate sizes increased, the AMN and AP contents gradually decreased. Furthermore, the enhanced levels of amino acids were observed under loquat, waxberry, and citrus intercropping in spring, which increased by 27.98%, 27.35%, and 26.21%, respectively. The contents of tea polyphenol and caffeine were lower under loquat and citrus intercropping in spring. These findings indicated that intercropping fruit trees, specifically loquat and citrus, have immense potential in promoting the green and sustainable development of tea plantations.

Integration of network pharmacology, lipidomics, and transcriptomics analysis to reveal the mechanisms underlying the amelioration of AKT-induced nonalcoholic fatty liver disease by total flavonoids in vine tea.

Nonalcoholic fatty liver disease (NAFLD) is the main reason for chronic liver diseases and malignancies. Currently, there is a lack of approved drugs for the prevention or treatment of NAFLD. Vine tea (Ampelopsis grossedentata) has been used as a traditional Chinese beverage for centuries. Vine tea carries out several biological activities including the regulation of plasma lipids and blood glucose, hepato-protective function, and anti-tumor activity and contains the highest content of flavonoids. However, the underlying mechanisms of total flavonoids from vine tea (TF) in the attenuation of NAFLD remain unclear. Therefore, we investigated the interventions and mechanisms of TF in mice with NAFLD using an integrated analysis of network pharmacology, lipidomics, and transcriptomics. Staining and biochemical tests revealed a significant increase in AKT-overexpression-induced (abbreviated as AKT-induced) NAFLD in mice. Lipid accumulation in hepatic intracellular vacuoles was alleviated after TF treatment. In addition, TF reduced the hepatic and serum triglyceride levels in mice with AKT-induced NAFLD. Lipidomics results showed 32 differential lipids in the liver, mainly including triglycerides (TG), diglycerides (DG), phosphatidylcholine (PC), and phosphatidylethanolamine (PE). Transcriptomic analysis revealed that 314 differentially expressed genes were commonly upregulated in the AKT group and downregulated in the TF group. The differential regulation of lipids by the genes Pparg, Scd1, Chpt1, Dgkz, and Pla2g12b was further revealed by network enrichment analysis and confirmed by RT-qPCR. Furthermore, we used immunohistochemistry (IHC) to detect changes in the protein levels of the key proteins PPARγ and SCD1. In summary, TF can improve hepatic steatosis by targeting the PPAR signaling pathway, thereby reducing de novo fatty acid synthesis and modulating the glycerophospholipid metabolism.

Effects of green tea and roasted green tea on human responses.

Our objective was to elucidate the effects of tea consumption on refreshment and stress reduction/recovery through examining the multiple associations among factors such as various physiological responses and task performance. Participants included 20 healthy young men who performed a mental arithmetic task while 11 physiological responses were measured. The experiments were conducted twice under different beverage consumption conditions on separate days. The mental arithmetic task was executed six times in 1 day; participants ingested hot water, green tea, or roasted green tea (hojicha) before each task. Several subjective assessments: subjective fatigue, stress, mental workload, and flow were evaluated after each task. The R-R intervals, heart rate variability spectral components, the Poincaré plot indices (SD1 and SD2) and plethysmogram amplitude tended to decrease during task periods compared to resting periods. Tissue blood volume/flow (TBV, TBF) and near-infrared spectroscopy responses (NIRS) were lower in the tea condition than in the hot water condition. By scrutinizing various indicators, we found that aromatic stimulation of Japanese tea beverages has the potential to induce positive effects, enhance mental task performance, promote refreshment, and alleviate feelings of fatigue. These positive effects were observed even in small quantities and within a short duration, mirroring responses observed in daily consumption.

Sustained oxidation of Tea-Fe(III)/H2O2 simultaneously achieves sludge reduction and carbamazepine removal: The crucial role of EPS regulation.

Establishing an economic and sustained Fenton oxidation system to enhance sludge dewaterability and carbamazepine (CBZ) removal rate is a crucial path to simultaneously achieve sludge reduction and harmless. Leveraging the principles akin to "tea making", we harnessed tea waste to continually release tea polyphenols (TP), thus effectively maintaining high level of oxidation efficiency through the sustained Fenton reaction. The results illustrated that the incorporation of tea waste yielded more favorable outcomes in terms of water content reduction and CBZ removal compared to direct TP addition within the Fe(III)/hydrogen peroxide (H2O2) system. Concomitantly, this process mainly generated hydroxyl radical (•OH) via three oxidation pathways, effectively altering the properties of extracellular polymeric substances (EPS) and promoting the degradation of CBZ from the sludge mixture. The interval addition of Fe(III) and H2O2 heightened extracellular oxidation efficacy, promoting the desorption and removal of CBZ. The degradation of EPS prompted the transformation of bound water to free water, while the formation of larger channels drove the discharge of water. This work achieved the concept of treating waste with waste through using tea waste to treat sludge, meanwhile, can provide ideas for subsequent sludge harmless disposal.

Dynamic changes in the non-volatile and flavour compounds in withered tea leaves of three different colour cultivars based on multi-omics.

In this study, metabolomics and proteomics were performed to investigate the fluctuations of non-volatile compounds and proteins in tea leaves from three tea cultivars with varying colours during withering. A total of 2798 compounds were detected, exhibiting considerable variations in amino acids, phenylpropanoids, and flavonoids. The ZH1 cultivar displayed increased levels of amino acids but decreased levels of polyphenols, which might be associated with the up-regulation of enzymes responsible for protein degradation and subsequent amino acid production, as well as the down-regulation of enzymes involved in phenylpropanoid and flavonoid biosynthesis. The FUD and ZH1 cultivars had elevated levels of flavanols and flavanol-O-glycosides, which were regulated by the upregulation of FLS. The ZJ and ZH1 cultivars displayed elevated levels of theaflavin and peroxidase. This work presents a novel investigation into the alterations of metabolites and proteins between tea cultivars during withering, and helps with the tea cultivar selection and manufacturing development.

Acid-catalyzed phenolation of lignin with tea polyphenol: Enhancing uv resistance and oxidation resistance for potential applications.

The rapid development of the industry has led to the destruction of the earth's ozone layer, resulting in an increasingly serious problem of excessive ultraviolet radiation. Exploring effective measures to address these problems has become a hot topic. Lignin shows promise in the design and preparation of anti-ultraviolet products due to its inherent properties. However, it is important to investigate way to enhance the reactivity of lignin and determine its application form in related products. In this study, phenolic reactions with tea polyphenols were conducted through acid-catalyzed conversion, utilizing organic solvent lignin as the primary material. The phenolic hydroxyl content of the original lignin increased significantly by 218.8 %, resulting in notable improvements in UV resistance and oxidation resistance for phenolic lignin. Additionally, micro-nanocapsule emulsions were formed using phenolic lignin particles as surfactants through ultrasonic cavitation with small-molecule sunscreens. A bio-based sunscreen was prepared with phenolated lignin micro-nanocapsules as the active ingredient, achieving an SPF 100.2 and demonstrating excellent stability. The sunscreen also exhibited strong antioxidant properties and impermeability, ensuring user safety. This research offers a current solution for improving the application of lignin in sunscreens while also broadening the potential uses of plant-based materials in advanced functional products.

Oral Antiviral Defense: Saliva- and Beverage-like Hypotonicity Dynamically Regulate Formation of Membraneless Biomolecular Condensates of Antiviral Human MxA in Oral Epithelial Cells.

The oral mucosa represents a defensive barrier between the external environment and the rest of the body. Oral mucosal cells are constantly bathed in hypotonic saliva (normally one-third tonicity compared to plasma) and are repeatedly exposed to environmental stresses of tonicity, temperature, and pH by the drinks we imbibe (e.g., hypotonic: water, tea, and coffee; hypertonic: assorted fruit juices, and red wines). In the mouth, the broad-spectrum antiviral mediator MxA (a dynamin-family large GTPase) is constitutively expressed in healthy periodontal tissues and induced by Type III interferons (e.g., IFN-λ1/IL-29). Endogenously induced human MxA and exogenously expressed human GFP-MxA formed membraneless biomolecular condensates in the cytoplasm of oral carcinoma cells (OECM1 cell line). These condensates likely represent storage granules in equilibrium with antivirally active dispersed MxA. Remarkably, cytoplasmic MxA condensates were exquisitely sensitive sensors of hypotonicity-the condensates in oral epithelium disassembled within 1-2 min of exposure of cells to saliva-like one-third hypotonicity, and spontaneously reassembled in the next 4-7 min. Water, tea, and coffee enhanced this disassembly. Fluorescence changes in OECM1 cells preloaded with calcein-AM (a reporter of cytosolic "macromolecular crowding") confirmed that this process involved macromolecular uncrowding and subsequent recrowding secondary to changes in cell volume. However, hypertonicity had little effect on MxA condensates. The spontaneous reassembly of GFP-MxA condensates in oral epithelial cells, even under continuous saliva-like hypotonicity, was slowed by the protein-phosphatase-inhibitor cyclosporin A (CsA) and by the K-channel-blocker tetraethylammonium chloride (TEA); this is suggestive of the involvement of the volume-sensitive WNK kinase-protein phosphatase (PTP)-K-Cl cotransporter (KCC) pathway in the regulated volume decrease (RVD) during condensate reassembly in oral cells. The present study identifies a novel subcellular consequence of hypotonic stress in oral epithelial cells, in terms of the rapid and dynamic changes in the structure of one class of phase-separated biomolecular condensates in the cytoplasm-the antiviral MxA condensates. More generally, the data raise the possibility that hypotonicity-driven stresses likely affect other intracellular functions involving liquid-liquid phase separation (LLPS) in cells of the oral mucosa.

Anti-Biofilm Activity of Assamsaponin A, Theasaponin E1, and Theasaponin E2 against Candida albicans.

Biofilm formation plays a crucial role in the pathogenesis of Candida albicans and is significantly associated with resistance to antifungal agents. Tea seed saponins, a class of non-ionic triterpenes, have been proven to have fungicidal effects on planktonic C. albicans. However, their anti-biofilm activity and mechanism of action against C. albicans remain unclear. In this study, the effects of three Camellia sinensis seed saponin monomers, namely, theasaponin E1 (TE1), theasaponin E2 (TE2), and assamsaponin A (ASA), on the metabolism, biofilm development, and expression of the virulence genes of C. albicans were evaluated. The results of the XTT reduction assay and crystal violet (CV) staining assay demonstrated that tea seed saponin monomers concentration-dependently suppressed the adhesion and biofilm formation of C. albicans and were able to eradicate mature biofilms. The compounds were in the following order in terms of their inhibitory effects: ASA > TE1 > TE2. The mechanisms were associated with reductions in multiple crucial virulence factors, including cell surface hydrophobicity (CSH), adhesion ability, hyphal morphology conversion, and phospholipase activity. It was further demonstrated through qRT-PCR analysis that the anti-biofilm activity of ASA and TE1 against C. albicans was attributed to the inhibition of RAS1 activation, which consequently suppressed the cAMP-PKA and MAPK signaling pathways. Conversely, TE2 appeared to regulate the morphological turnover and hyphal growth of C. albicans via a pathway that was independent of RAS1. These findings suggest that tea seed saponin monomers are promising innovative agents against C. albicans.