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IntroductionAntisense oligonucleotides (ASOs) with therapeutic potential have recently been reported to target the SARS-CoV-2 genome. Peptide nucleic acids (PNAs)-based ASOs have been regarded as promising drug candidates, but intracellular delivery has been a significant obstacle. Here, we present novel modified PNAs, termed OPNAs, with excellent cell permeability that disrupt the RNA genome of SARS-CoV-2 and HCoV-OC43 by introducing cationic lipid moiety onto the nucleobase of PNA oligomer backbone.MethodsHCT-8 cells and Caco-2 cells were treated with 1 μM antisense OPNAs at the time of viral challenge and the Viral RNA levels were measured by RT-qPCR three days post infection.ResultsNSP 14 targeting OPNA 5 and 11, reduced the viral titer to a half and OPNA 530, 531 and 533 lowered viral gene expression levels to less than 50% of control by targeting the 5’ UTR region. Several modifications (oligo size and position, etc.) were introduced to enhance the efficacy of selected OPNAs. Improved OPNAs exhibited a dose-dependent reduction in viral replication and nucleoprotein (NP) protein. When a mixture of oligomers was applied to infected cells, viral titer and NP levels decreased by more than eightfold.DiscussionIn this study, we have developed a modified PNA ASO platform with exceptional chemical stability, high binding affinity, and cellular permeability. These findings indicate that OPNAs are a promising platform for the development of antivirals to combat future pandemic viral infections that do not require a carrier.

Callus suspension techniques have been considered attractive for improving bioactive metabolite productivity; methyl jasmonate (MeJA) is a widely used elicitor for stimulating synthetic pathways. In this study, a multivariate analysis-based metabolomics approach was employed to investigate the primary and specialized metabolites in the leaves, unelicited calli, and 100 or 200 μM MeJA elicited calli of Damnacanthus major. Rubiadin, a powerful anthraquinone with various therapeutic properties, was only identified in D. major calli, accumulating in a MeJA elicitation concentration-dependent manner. Callus cultures also contained high levels of amino acids, sugars, and phenolic compounds, indicating energy metabolism and metabolic adaptation responses for proliferation and stabilization. Regarding MeJA application, elicited calli contained higher amounts of quinic acid, kaempferol, and glucose with lower amounts of sucrose and raffinose than those in the unelicited control, which were closely related to protective mechanisms against MeJA. Moreover, excessive elicitation increased the asparagine, fructose, and raffinose levels and decreased the glucose and sucrose levels, which was ascribed to increased activation of the aminoacyl-tRNA biosynthesis pathway and wider utilization of glucose than of fructose after sucrose degradation. These results will be useful for optimizing plant cell culture techniques to achieve high production rates for valuable specialized metabolites.

In Jeju-native Citrus, flavonoids are the main contributors to the various types of biological activity, such as antioxidant, antitumor, and anti-inflammatory activity. Thus, we developed simultaneous quantification methods for the analysis of ten bioactive flavonoids in Jeju Citrus fruits (Dangyuja, Gamja, Jigak, Sadugam, and Soyuja) harvested at six different time points using a high-performance liquid chromatography–diode array detector (HPLC-DAD). Separation was performed using a flow rate of 0.8 mL/min, a column temperature of 40 °C, a mobile phase buffer of 0.5% acetic acid, and a detection wavelength of 278 nm. The established analytical method showed good linearity (R2 ≥ 0.9997), precision (inter-day < 0.599%, intra-day < 0.055%), and accuracy (recoveries 92.30–108.80%). The HPLC–DAD method was subsequently applied to analyze flavonoids in Citrus samples. Overall, the quantification results indicated that the compositions and content of flavonoids differed for each Citrus species. The harvesting period also influenced the changes in flavonoid content within each Citrus species. The analytical results with chemometrics revealed that higher flavonoid levels in early-harvested Citrus were derived from the improved fruit size and reduced flavonoid synthesis during maturation. This study provides a practical and reliable method for the analysis of ten flavonoids that can be further utilized in the quality assessment of Jeju Citrus.

Current research highlights the use of natural products or phytochemicals as drugs and functional additives to treat obesity with few side effects. Sargassum horneri (SH) and Ulva australis (UA) are marine waste resources on Jeju Island, Republic of Korea. In this study, we analyzed their antioxidant and anti-obesity efficacies to confirm their potential as functional additives. We prepared SH and UA extracts using 80% ethanol and observed that free radical scavenging activity and total phenol content were high in SH extracts, and total flavonoid content was elevated in UA extracts. Additionally, 3T3-L1 cells were treated with SH and UA extracts, and the ability of the extracts to inhibit adipocyte differentiation was examined using Oil Red O staining and analysis of neutral fat content. We confirmed that the mRNA expression of the C/EBPα, PPARγ, and SREBP1c genes that act on adipocyte differentiation, and of FAS, a fatty acid synthase, was suppressed. Experiments in a mouse model of obesity showed that 12-week administration of a high-fat diet with 1% extract added to drinking water resulted in lower weight gain compared to the high-fat diet alone. These results suggest that SH and UA extracts have antioxidant properties and are effective in obesity prevention. Therefore, the two marine waste resources are potential functional additive candidates for preventing obesity.

Magnolia flower buds are a source of herbal medicines with various active compounds. In this study, differences in the distribution and abundance of major essential oils, phenolic acids, and primary metabolites between white flower buds of Magnolia heptapeta and violet flower buds of Magnolia denudata var. purpurascens were characterised. A multivariate analysis revealed clear separation between the white and violet flower buds with respect to primary and secondary metabolites closely related to metabolic systems. White flower buds contained large amounts of monoterpene hydrocarbons (MH), phenolic acids, aromatic amino acids, and monosaccharides, related to the production of isoprenes, as MH precursors, and the activity of MH synthase. However, concentrations of β-myrcene, a major MH compound, were higher in violet flower buds than in white flower buds, possibly due to higher threonine levels and low acidic conditions induced by comparatively low levels of some organic acids. Moreover, levels of stress-related metabolites, such as oxygenated monoterpenes, proline, and glutamic acid, were higher in violet flower buds than in white flower buds. Our results support the feasibility of metabolic profiling for the identification of phytochemical differences and improve our understanding of the correlated biological pathways for primary and secondary metabolites.

Objective We aimed to develop an International Classification of Diseases (ICD) 10th edition injury code-based disability-adjusted life year (DALY) to measure the burden of specific injuries. Methods Three independent panels used novel methods to score disability weights (DWs) of 130 indicator codes sampled from 1,284 ICD injury codes. The DWs were interpolated into the remaining injury codes (n=1,154) to estimate DWs for all ICD injury codes. The reliability of the estimated DWs was evaluated using the test-retest method. We calculated ICD-DALYs for individual injury episodes using the DWs from the Korean National Hospital Discharge Injury Survey (HDIS, n=23,160 of 2004) database and compared them with DALY based on a global burden of disease study (GBD-DALY) regarding validation, correlation, and agreement for 32 injury categories. Results Using 130 ICD 10th edition injury indicator codes, three panels determined the DWs using the highest reliability (person trade-off 1, Spearman r=0.724, 0.788, and 0.875 for the three panel groups). The test-retest results for the reliability were excellent (Spearman r=0.932) (P

Purpose: To investigate skin care efficacies of Zanthoxylum pipetitum extract and isolated compounds. Methods: Ethanol extracts of leaves, branches and fruits of what were partitioned into n-hexane, chloroform, ethyl acetate, n-butanol and aqueous layers and some fractions were further analyzed to isolate five compounds. The isolated compounds were identified based on the proton and carbon nuclear magnetic resonance (NMR) spectra. Cosmetic efficacy tests of the extracts and isolated compounds were evaluated by in vitro tests. Results: Phytochemical studies of the chloroform and ethyl acetate layers led to the isolation of five compounds; quercitrin (1), afzelin (2), hydroxy-a-sanshool (3), a-sanshool (4) and hyperoside (5). In activity tests, the extracts showed inhibitory activity against inflammation response and melanin synthesis, and induction of procollagen type I C-peptide (PIP). Among the isolated compounds, hydroxy-a-sanshool (3) and a-sanshool (4) displayed significant anti-inflammatory activity. Conclusion: The results demonstrate that Z. piperitum extract and its active compounds possess a significant potential as a cosmeeutical agent for enhancing skin quality. [ABSTRACT FROM AUTHOR]

Sargassum horneri (SH) and Ulva australis (UA) are marine waste resources that cause environmental and economic problems when entering or multiplying the coastal waters of Jeju Island. We analyzed their anti-diabetic efficacy to assess their reusability as functional additives. The alpha-glucosidase inhibitory activity of SH and UA extracts was confirmed, and the effect of UA extract was higher than that of SH. After the induction of insulin-resistant HepG2 cells, the effects of the two marine extracts on oxidative stress, intracellular glucose uptake, and glycogen content were compared to the positive control, metformin. Treatment of insulin-resistant HepG2 cells with SH and UA resulted in a concentration-dependent decrease in oxidative stress and increased intracellular glucose uptake and glycogen content. Moreover, SH and UA treatment upregulated the expression of IRS-1, AKT, and GLUT4, which are suppressed in insulin resistance, to a similar degree to metformin, and suppressed the expression of FoxO1, PEPCK involved in gluconeogenesis, and GSK-3β involved in glycogen metabolism. The oral administration of these extracts to rats with streptozotocin-induced diabetes led to a higher weight gain than that in the diabetic group. Insulin resistance and oral glucose tolerance are alleviated by the regulation of blood glucose. Thus, the SH and UA extracts may be used in the development of therapeutic agents or supplements to improve insulin resistance.