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Thlaspi arvense Linn, known as “Pennycress,” is a plant from the Brassicaceae family that has been traditionally used to treat glomerulonephritis, gastritis, and rheumatoid arthritis. As part of our current projects to discover bioactive compounds from natural resources, three flavonoids (1-3), one diarylheptanoid (4), and two phenolic glycosides (5-6) were isolated from the whole plant of T. arvense via semi-preparative HPLC purification. The chemical structures of compounds 1-6 were elucidated as isoorientin (1), isovitexin (2), isoscoparin (3), oregonin (4), 1,2-disinapoylgentiobiose (5), and 1,2,2’-trisinapoylgentiobiose (6) based on the comparison of their NMR spectroscopic and physical properties with those of previous studies. Notably, this is the first report of the presence of compounds 3-6 in this plant. Compounds 1-6 were then tested to determine their effects on osteogenesis and adipogenesis in the mouse mesenchymal stem cell (MSC) line C3H10T1/2. Among the six compounds, isovitexin (2) was found to promote the osteogenic differentiation of MSCs. As the concentration of 2 increased, the differentiation of MSCs into osteoblasts became more active, as evidenced by the induction of mRNA expression of the osteogenic markers, alkaline phosphatase (ALP) and osteopontin (OPN). Accordingly, our findings demonstrate that isovitexin (2) could potentially serve as a valuable compound for the treatment of menopause-associated syndromes, such as osteoporosis, by promoting MSC osteogenesis.

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Macrophage plays a critical role in inducing inflammatory response, activated though TLR4 receptor by endotoxin lipopolysaccharide (LPS). It has been reported that cell inflammation is related to the oxidative stress response. However, it is not clear how the LPS-induced inflammatory signaling pathway is mechanistically related to oxidative stress responses. Here, we examined the impact of LPS-induced inflammation and oxidative stress using RAW 264.7 cells. LPS treatment up to 1 µg/ml did not show any cell toxicity. LPS stimulation causes Akt and ERK1/2 activation. However, treatment with LY294002, an Akt signaling inhibitor, completely inhibited Akt activation but not ERK1/2 activation. Interestingly, We found that LPS induces down-regulation of Nrf2 nuclear expression and translocation, whereas inhibition of Akt activation with LY294002 reverses the effects of LPS on Nrf2, as judged by Western blotting with nuclear fraction protein and immunofluorescence analysis. In addition, to investigate the role of Akt signaling in NF-κB activation-induced inflammatory response, NF-κB nuclear translocation and its target gene expression were analyzed by immunofluorescence assay and qRT-PCR analysis. LPS-induced NF-κB nuclear translocation was suppressed by LY294002 treatment. Furthermore, LY294002 inhibited LPS-induced mRNA expression of inflammatory markers, including IL-1β and MCP-1. In summary, these findings suggest that Akt signaling plays a critical role in the LPS-induced inflammatory and oxidative stress responses in RAW 264.7 cells.

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Appropriate crystal growth rate is a critical factor in obtaining a high-quality crystal. Traditionally, optimizing crystal growth rate is usually through increasing or decreasing protein and precipitant concentration. Here, a method will be shown to control nucleation rate and increase crystal size. By adding additional water to crystal drop we can decrease protein and precipitant concentration at an equal proportion, which allows a lower supersaturate environment at the initial stage of crystallization. For this reason, over-nucleated protein molecules can only bind to the surface of a limited number of growing centers. Then, the additional water will leave a crystal drop in the reservoir solution after equilibration and does not change the final ratio of protein and precipitant reagent. As a result, the total crystal number decreases but the crystal size has a dramatic increase, as well as the morphology of crystals. In the current study, pleckstrin homology (PH) domain of 3-phosphoinositide-dependent protein kinase 1 protein (PDK1) was taken as an example to illustrate this phenomenon and its underlying mechanism, which aims to provide a strategy for crystal optimization.

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This study aimed to investigate the pharmacokinetics of curcumin (CUR) and its anti-cancer activity against B16F10 cell in mice using CUR- and pluronic P85 (PP85)-loaded mesoporous silica nanoparticles. CUR was incorporated into mesoporous nanosilicate (SMB7) using a solvent extraction method, and PP85 was also loaded into SMB7 using a freeze-drying method. The final formulation, CUR-PP85-SMB7, was optimized at a ratio of 1:2:4 (w/w). The solubility and release profile of CUR from CUR-PP85-SMB7 were greatly improved compared with CUR alone. The plasma concentrations of CUR were also increased in mice administered CUR-PP85-SMB7 intraperitonially at a dose of 5 mg CUR compared with equivalent dose of standalone CUR. Moreover, tumor growth was inhibited through the repeated intraperitonial administration of CUR-PP85-SMB7 (20 mg CUR equivalent/kg for 7 days) in B16F10 melanoma-bearing mice, and the inhibition was even more marked than that of the CUR-SMB7 group with equivalent dose and the same dose in the CUR-only treatment group. The CUR concentration in tumor tissues at 4 h after the last CUR dose was the highest in the CUR-PP85-SMB7 group, and comparable in the CUR-SMB7 and CUR-only groups. These findings suggest that the P-glycoprotein (P-gp) inhibitory effect of PP85 from CUR-PP85-SMB7 can increase the CUR accumulation in B16F10 melanoma tissues and improved the anti-cancer efficacy. In conclusion, the improved pharmacokinetic properties and anti-cancer activity of CUR could be achieved by preparing CUR- and PP85-loaded mesoporous silica nanoparticles. Incorporation of PP85, a surfactant that has P-gp inhibitory effect, into SMB7 together with CUR potentiated the anti-cancer effect of CUR in B16F10 melanoma-bearing mice by increasing the release and the intratumor concentration of CUR.

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Detomidine, an α2-adrenergic receptor agonist, is widely employed in horses due to its effective sedative and analgesic properties. The International Federation of Horseracing Authorities prohibits the administration of detomidine during periods that may affect racing performance. It has established the International Screening Limits in plasma at 0.02 ng/mL of its metabolites, 3-hydroxy detomidine. The biotransformation of detomidine involves aliphatic hydroxylation, generating 3-hydroxy detomidine, which subsequently undergoes an additional dehydrogenation reaction to produce 3-carboxy detomidine. Therefore, the detection of 3-carboxy detomidine following 3-hydroxy detomidine suggests a potential enhancement in doping control detection capability. In this study, we investigated the pharmacokinetics of detomidine, and its metabolites following intravenous administration and monitored drug profiles in plasma to prevent substance abuse for doping control. For assessment, the LC-MS/MS method was employed, and the validity of the analytical method was evaluated. The results demonstrated detectable concentrations of detomidine above the lower limit of quantification for 8 hours, and both metabolites were consistently detected throughout the experimental period (48 hours). Monitoring both 3-hydroxy detomidine and 3-carboxy detomidine is considered advantageous for detomidine abuse control. In particular, the calculated long half-life of 3-carboxy detomidine demonstrates its potential as a substance useful for doping tests.

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Lower respiratory tract infections are prevalent in South Korea, and clarithromycin, amoxicillin, clavulanate, and loxoprofen are frequently prescribed for these infections. In addition, co-administration of these small molecule drugs and Korean traditional medicine (KTM) are common. Despite a growing number of patients administering these drugs in combination, their drug-drug interaction (DDI) potentials have not been examined. Therefore, our goal was to assess the potential DDIs among clarithromycin, amoxicillin, clavulanate, and loxoprofen when administered together using physiologically based pharmacokinetic (PBPK) modeling approach. The PBPK models of the drugs were constructed and validated based on the previously reported studies using SimCYP^TM. Subsequently, the DDI potentials were evaluated in scenarios with one drug as the substrate (clarithromycin 250 mg, amoxicillin 250 mg, clavulanate 125 mg, and loxoprofen 60 mg) and the others as inhibitors. The simulated pharmacokinetic parameters were validated against observed data, showing good agreement within predefined criteria (0.5-2.0). The observed data fell within the 5th to 95th confidence interval of the simulated data, indicating accurate description of plasma concentration-time profiles. Assessing DDI potentials through AUC ratios (with inhibitors/without inhibitors), the predicted AUC ratios ranged from 0.89 to 1.03, indicating no significant DDIs (criteria: 0.80-1.25). In summary, our study supports the safe use of a combination of clarithromycin, amoxicillin, clavulanate, and loxoprofen.

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Atorvastatin and amlodipine are commonly prescribed for the treatment of cardiovascular diseases, which often necessitate a polypharmacy approach due to long-term and combined medication use, thus increasing the potential for drug-drug interactions (DDIs). In this study, we developed physiologically-based pharmacokinetic (PBPK) models for atorvastatin and amlodipine, and simulated the potentials of DDIs using known strong index perpetrators of metabolic enzymes, including itraconazole, clarithromycin, and rifampicin. These PBPK models were developed using PK-Sim^® and a virtual Asian Population was developed based on Asian Individuals within PK-Sim^® without modification. Model validation was achieved by comparing simulated exposures to observed data, and all input parameters underwent local sensitivity analysis. We simulated the pharmacokinetic profiles of atorvastatin and amlodipine both with and without the presence of itraconazole, clarithromycin, and rifampicin, and investigated the geometric mean ratios (GMRs) of AUC_inf and C_max for each drug. The GMRs of AUC_inf for atorvastatin were 2.64, 2.37, and 0.43 with itraconazole, clarithromycin, and rifampicin, respectively; for amlodipine, the GMRs of AUC_inf were 2.06, 1.69, and 0.27, respectively. These results demonstrate that our models can be effectively used to explore further the potential DDIs of atorvastatin and amlodipine with various comedications.

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Asthma, a prevalent chronic inflammatory lung disease that affects over 330 million people worldwide, manifests through symptoms such as wheezing, coughing, dyspnea, and chest tightness. Probiotics, such as Bacillus subtilis (B. subtilis) and Lactobacillus rhamnosus (L. rhamnosus), provide protection against allergic airway inflammation. This is achieved by the induction of regulatory immune responses and restoration of cytokine balance in activated immune cells. This study aimed to investigate the protective effects of probiotics against inflammation in pulmonary diseases by assessing their impact on airway resistance, immune cell infiltration, and allergy-related chemokine levels in an ovalbumin (OVA)-induced asthmatic mouse model. Oral administration of live L. rhamnosus and B. subtilis significantly decreased airway resistance and reduced infiltration of immune cells like eosinophils and macrophages. Histological assessment demonstrated the anti-inflammatory effects of B. subtilis and L. rhamnosus. Furthermore, both probiotics could decrease the production of allergic factors, such as interleukin-13 (IL-13) and immunoglobulin (Ig) E and IgG1. IL-13 reduction by B. subtilus and L. rhamnosus also led to decreasing mucus production. This study demonstrated that oral administration of probiotics from Korean fermented foods may induce potentially preventive effects against asthmatic lung inflammation.

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Organoids are cell aggregates made by reprocessing stem cells through a three-dimensional culture method and are expected to revolutionize biomedical research and clinical treatment. However, organoid regulation is unclear and has ethical issues that remain unresolved. We performed to systematically reviewed the regulation of organoids internationally, detailing the definition of organoids, and their regulation in ethical and technical aspects. In this cross-sectional study, we systematically searched the official websites of regulatory authorities in the European Union, United Kingdom, United States, Canada, Japan, South Korea, Switzerland, Germany, China, Netherlands, and International Society for Stem Cell Research (ISSCR) up to May 2023. However, if no organoid regulations are published on the official worldwide website, regulations are looked into in the stem cell and embryo domains. Organoid regulations were extracted that covered informed consent, chimeras and gastruloids, germ cells, and comparisons were made between countries. The only ISSCR has standard, formal definitions for organoids, there was no document describing the definition of organoid in any other legislation or regulations. In organoid regulations on ethics, several national laws restrict organoid research. Also, replacing organoids with animal testing has been overall positive in the investigated countries. Research on organoids is advancing rapidly. To build science-based research ethics and laws, organoid regulations call for the creation of convergence grounds, it is crucial to keep moving toward the goal of broad convergence in the field of common research ethics.

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Little is known on the real-world comparative effectiveness of sodium-glucose cotransporter-2 inhibitors (SGLT2i) versus angiotensin receptor-neprilysin inhibitor (ARNi) used for heart failure (HF) management. This study used South Korea’s nationwide claims data from 2015 to 2020 to construct a population-based cohort of new users of SGLT2is or ARNi. Individuals were followed from the first prescription date of SGLT2is or ARNi until outcome occurrence, treatment switch or discontinuation, death, or end of the study period. Within the 1:1 propensity score-matched cohort, we estimated hazard ratios (HR) with 95% confidence intervals (CI) for the risk of HF admission with SGLT2is compared with ARNi using proportional subdistribution hazards model of Fine and Gray. We identified 496 propensity-score matched patient-pairs of SGLT2is and ARNi; with a mean age of 72.5 years and a male representation of 57.6%. Incidence rate of HF admission was 27.3 and 35.6 per 100 person-years in SGLT2is and ARNi group. When comparing the risk of HF admission associated with SGLT2is group with ARNi group, HR was 0.71 (95% CI 0.48-1.04). Effect modifications were observed by history of hospitalization for HF (p-for-interaction=0.002) and by recent use of renin-angiotensin-system inhibitors (p-for-interaction= 0.005). With future studies using more recent data warranted to corroborate our study results, these preliminary findings support current guideline recommendations for HF management and further, suggest similar effectiveness between SGLT2is and ARNi in routine care settings.

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Exosomes are extracellular vesicles surrounded by a lipid bilayer released from various types of cells, including normal and cancer cells. Exosomes contain a variety of molecules, such as nucleic acids, proteins, and lipids, which are transported to other cells and mediate intercellular communication. Tumor-derived exosomes have different cargos than normal exosomes. Tumor-derived exosomes promote cancer growth, progression, metastasis, and angiogenesis. In addition, tumor-derived exosomes mediate tumor immune escape by regulating the activity of immune cells in tumor microenvironments. The significance of tumor-derived exosomes extends beyond their biological roles; they hold immense potential as clinical tools. Recently, tumor-derived exosomes have been used as biomarkers in liver cancer and prostate cancer, offering a non-invasive method for early detection and monitoring of these malignancies. Moreover, numerous studies have explored the use of exosomes for drug delivery, highlighting their capability to enhance the efficacy and reduce the side effects of conventional therapies and this advancement potentially transforms cancer treatment paradigms. This review discusses exosome biogenesis and the role of exosome cargo components, emphasizing their critical functions in intercellular communication within the tumor microenvironment. Furthermore, it delves into the innovative applications of exosomes as biomarkers for cancer diagnosis and as vehicles for targeted drug delivery, underscoring their importance in advancing cancer research and therapy.

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Neuromyelitis optica spectrum disorder (NMOSD) is an infrequent inflammatory autoimmune disorder of the central nervous system, that affects the spinal cord and optic nerves. Aquaporin 4 IgG (AQP4-IgG) is a seropositive-specific antibody detected in the serum of a patient with NMO. AQP4-IgG binds to AQP4 via the astrocyte end-feet, causing a classical complement cascade that results in an inflammatory response-induced astrocyte injury and a secondary response involving oligodendrocyte loss and demyelination. NMO causes severe relapses and affects women more than men. A patient with NMO experiences pain primarily as transverse myelitis with longitudinally extensive recurrences or bilateral optic neuritis. Treating NMOSD poses considerable clinical challenges. This review provides an overview of the state-of-the-art studies summarizing the pain, prevalence, and primary pathology of NMOSD and discusses several potential therapeutic targets for its treatment.