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Estrogen receptor-α (ERα) is a 66 kDa (ERα66) nuclear receptor transcription factor and it functionally plays a crucial role in various processes associated with human breast cancer. 36 kDa novel variant of ERα66 has been identified and cloned, and named as ERα36. As reported previously, we observed a downregulation of ERα66 expression and an elevation of ERα36 levels in tamoxifen-resistant breast cancer (TAMR-MCF-7) compared to the parental MCF-7 cells. We investigated the functional roles of ERα66 and ERα36 and their potential mechanisms in regulating the epithelial-mesenchymal transition (EMT) process. Our findings revealed that the upregulation of ERα36 led to the downregulation of ERα66, consequently contributing to the acquisition of EMT and promoting metastasis. Furthermore, we explored the involvement of a long non-coding RNA (lncRNA) called homeobox transcript antisense intergenic RNA (HOTAIR) in the induction and regulation of EMT during metastasis. Interestingly, we observed that the increased expression of ERα36 resulted in elevated HOTAIR levels, while upregulation of ERα66 or downregulation of ERα36 abolished HOTAIR expression. Functional depletion of HOTAIR significantly impaired EMT and cell migration in TAMR-MCF-7 cells. Collectively, our data demonstrate that ERα36 acts as a key regulator of EMT-driven metastasis in human breast cancer by directly modulating the long non-coding RNA HOTAIR.

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Cyclic GMP-AMP synthase (cGAS) plays an important role in inducing innate immune responses necessary for defense against invading pathogens and for repair of internal tissue damage. However, excessive accumulation of intracellular double-stranded DNA (dsDNA) leading to overactivation of cGAS results in a variety of chronic diseases including autoimmune disorders and inflammatory diseases. Therefore, a specific cGAS inhibitor can be utilized as an efficient therapeutic strategy for the prevention or treatment of chronic inflammatory diseases. We found that butaclamol inhibited cGAS activation induced by dsDNA by IFN-stimulated response elements-dependent luciferase reporter assay. Butaclamol reduced dsDNA-induced generation of 2’3’-cGAMP, of which production is dependent on cGAS enzymatic activity. Butaclamol did not suppress a STING agonist-induced luciferase reporter gene expression. The results suggest that butaclamol inhibits cGAS, but not STING. The inhibition of cGAS activation by butaclamol culminated in the reduction of cGAS-target gene, IFN-β. Butaclamol did not inhibit IFN-β production induced by agonists of other pattern-recognition receptors, suggesting that the inhibitory effect of butaclamol is rather specific for cGAS. Our results present butaclamol as a novel cGAS inhibitor and further suggest the possibility for therapeutic potential of butaclamol in prevention and treatment of cGAS-related immune diseases.

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Irisin is a myokine primarily expressed in skeletal muscle, known to mediate the beneficial effects of exercise. Irisin was first reported to induce adipocyte browning by increasing the expression of uncoupling protein 1 (UCP1), and subsequent studies have also discovered anti-oxidant, anti-inflammatory, and anti-metastatic effects of irisin. Despite controversies on the circulating levels of irisin, studies that tested the therapeutic potential of irisin by injecting recombinant irisin have shown promising results. Therefore, there is a need to develop means to construct a high-yield recombinant strain with optimal human irisin coding sequence and proper glycosylation. In this study, we cloned and expressed human irisin (r-irisin) in Nicotiana benthamiana leaves and tested its biological activity in cultured adipocytes. Results showed that we have successfully constructed, expressed, and purified the glycated form of human irisin using N. benthamiana. Upon treatment with plant-made r-irisin on mature adipocytes, expression of UCP1 as well as mitochondrial biogenesis-related genes were increased, while lipid accumulation was decreased. In addition, plant made r-irisin treatment resulted in inhibition of adipogenesis evidenced by downregulated C/EBPα, PPARγ, and FABP4 expression and reduced lipid accumulation, implicating their therapeutic potential in the treatment of obesity-induced metabolic diseases. In summary, we developed a safe, efficient, cost-effective means for the production of human irisin in plants and demonstrated its biological activity in adipocytes.

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Cytokine-induced killer (CIK) cells are a heterogeneous population typically including more than 30% of CD3⁺CD56⁺ cells and efficiently kill tumor cells. Here, we provide important technical tips to generate successfully CIK cells from mouse spleen cells. To keep the viability and proliferation of CIK cells, two critical factors were essential: the consecutive addition of IFN-γ, anti-CD3 antibody, and a large enough amount of IL-2; cell density adjustment during the cultivation. On day 14, the cell number increased by > 200-fold, the phenotypes were > 90% CD3⁺CD8⁺ and > 45% CD3⁺NK1.1⁺. They expressed IFN-γ, IL-2, granzymes, and FasL, and had strong antitumor activity in vitro and in vivo. Taken together, our data provide an optimal protocol for the generation of CIK cells having more than > 30% of CD3⁺CD56⁺ cells.

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Glycyrrhizae Radix (GR) is a widely used herbal medicine. Its pharmacological efficacy depends largely on the composition of bioactive saponins and flavonoids, including glycyrrhizin, liquiritin, isoliquiritin, isoliquiritigenin, and liquiritigenin. This study aimed to compare the pharmacokinetics of glycyrrhizin, liquiritin, isoliquiritin, isoliquiritigenin, and liquiritigenin between orally administered GR extract and an equal amount of a single component. This study also aimed to investigate the intestinal absorption and metabolism of these major pharmacological components in rats. Plasma concentrations of liquiritigenin and isoliquiritigenin from the administered GR extract rapidly decreased for 4 h, increased for 4-10 h, and subsequently stabilized. However, a different profile was observed following a single administration of liquiritigenin or isoliquiritigenin despite a similar dose (0.5 mg/kg or 0.2 mg/kg, respectively). In contrast, the plasma concentrations and pharmacokinetic parameters of glycyrrhizin, liquiritin, and isoliquiritin differed insignificantly from the corresponding results of equivalent doses in rats. Consistent with the pharmacokinetic results, the apparent permeability of liquiritigenin and isoliquiritigenin from the administered GR extract increased by 2.2- and 4.8-fold, respectively, compared with that of individual components. Additionally, isoliquiritigenin was formed from isoliquiritin at the highest rate, converted from liquiritigenin at the lower rate, and converted from liquiritin at the lowest rate in intestinal segments of the rats. Liquiritigenin had a similar process. Therefore, isoliquiritin, liquiritin, and liquiritigenin could be biosources for isoliquiritigenin and liquiritigenin, which could occur in intestinal enterocytes, and the converted metabolites are absorbed into the plasma. Conclusively, the beneficial interaction between liquiritigenin and isoliquiritigenin in orally administered GR extract via metabolic conversion in intestinal enterocytes and enhanced absorption could provide a basis for treatment with GR extract rather than with the individual components.

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We aimed to evaluate the cost-effectiveness of the cytochrome p450 (CYP) 2C19 genotype-guided antiplatelet therapy compared with using clopidogrel or prasugrel without considering genotypes in patients with acute coronary syndrome undergoing percutaneous coronary intervention in Korea. We performed a cost-effectiveness analysis using a decision analytic model with a one-year time period employing a societal perspective. A cohort of 10,000 patients was assigned to each treatment alternative as follows: (1) selecting antiplatelet therapy between clopidogrel and prasugrel utilizing the genotype information of CYP2C19; (2) using clopidogrel without using genotype information; (3) using prasugrel without using genotype information. The primary outcome measure was the incremental cost per effectiveness ratio (ICER), where the effectiveness was defined as a death or cardiovascular event avoided, comparing genotype-guided therapy with the other two alternatives. The cardiovascular event was defined as any occurrence of nonfatal myocardial infarction, nonfatal stroke, and bleeding. The secondary outcome measures were the incidence of each event for each alternative and the number needed to treat (NNT) with the genotype-guided therapy versus each of other alternatives. The treatment strategy of genotype-guided therapy was dominant when compared with using clopidogrel (ICER: −36,610,303 KRW, −33,282 USD) or prasugrel (ICER: −24,202,531 KRW, −22,002 USD) for all patients without considering the genotype. The NNT of the genotype-guided therapy versus clopidogrel and prasugrel for all patients was 26 and 12, respectively, for any occurrence of death and cardiovascular event. Genotype-guided antiplatelet therapy in patients with acute coronary syndrome appeared to be a cost-effective treatment strategy in Korean patients.

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Polyelectrolyte complexes (PICs) have emerged as promising candidates in the field of drug delivery. This manuscript presents a comprehensive investigation organized into four main sections to elucidate the potential of PICs for drug delivery applications. The first section provides a foundational understanding of polyelectrolytes, highlighting their unique properties and characteristics. Subsequently, we delve into an examination of the physicochemical properties of polyelectrolytes, shedding light on the mechanisms governing the formation of PICs and the critical parameters involved in preparing polyelectrolyte nanoparticles. In the third section, we explore the formation of polyelectrolyte-based nanoparticles, encompassing hydrophilic polyelectrolytes, hydrophobically modified polyelectrolytes, and PICs formed with oppositely charged polymers. This section offers insights into the versatile applications of PICs in drug delivery systems. The final section delves into the biological functions of polyelectrolytes, revealing their role in enhancing cellular uptake, facilitating endosomal escape, acting as immune stimulators, and exhibiting antitumor activity. These insights emphasize the potential of PICs to overcome barriers in drug delivery and enhance therapeutic outcomes. In summary, this manuscript serves as a comprehensive resource for researchers and practitioners in the field, offering a holistic understanding of the multifaceted applications of PICs in drug delivery.

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Neurotransmitters play an important role in the human body, as regulators and messengers in cellular function, body regulation, and homeostasis, operating through the nervous system and organs. As neurotransmitters can directly affect the central nervous system, including the brain, they are thought to be directly and/or indirectly related to various neurological disorders. Therefore, to understand the pathophysiological mechanisms underlying neurological disorders, analytical methods are required for detecting neurotransmitters in biological samples. Although numerous sample preparation techniques have been introduced for neurotransmitter profiling, advanced microextraction techniques are characterized by simple and easy operation, consuming less of harmful organic solvents and wasting fewer biological samples. Chromatography and mass spectrometry are indispensable for neurotransmitter profiling. Chromatography can separate various neurotransmitters from matrix interference in complicated biological samples. Mass spectrometry combined with chromatographic separation enables sensitive and selective determination of trace levels of neurotransmitters. Furthermore, several trials have been conducted to identify neurotransmitters as potential biomarkers for neurological diseases. In this study, we review and organize recently reported articles to help readers develop analytical methods for profiling neurotransmitters and describe applications to biological samples, to provide insights into the mechanisms of neurological diseases.

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Parkinson’s disease (PD) is characterized by a progressive loss of dopaminergic neurons, leading to a decline in dopamine levels and the manifestation of motor and non-motor symptoms. The current treatments primarily focus on symptom management, leaving the need for disease-modifying therapies unmet. Emerging research has shown that neuroinflammation plays a pivotal role in the pathogenesis and progression of PD. Dipeptidyl peptidase-4 (DPP-4) inhibitors, originally developed for diabetes treatment, represent a novel avenue of investigation in the context of PD treatment. Beyond their established role in glycemic control, DPP-4 inhibitors have shown promise in modulating inflammatory responses within the central nervous system. This indicates their potential application in controlling neuroinflammation in PD, which in turn, could impact disease progression and neurodegeneration. This review summarized the current understanding of the role of DPP-4 inhibitors in neuroinflammation and their potential to modulate PD progression. Understanding the intricate interplay between neuroinflammation and dopaminergic neuron degeneration could pave the way for a new era of disease-modifying strategies, and DPP-4 inhibitors could offer a potential avenue to address both inflammation and neurodegeneration in PD.