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This file contains Supplemental figures, tables, and legends that accompany the main figures, as indicated in each legend. There are 3 supplemental tables and 6 supplemental figures. Table S1. List of potential target of the GNS compounds that were profiled in Figure 1 of the main text. Table S2. Half-maximal inhibitor concentration of the EGFR inhibitors tested. Table S3. Blood-brain-barrier penetration of the GNS compounds as assessed by the indicated pharmacokinetic measurements. Figure S1. Structural modeling studies show the binding characteristics of the indicated EGFR inhibitors. Figure S2. The effects of each EGFR inhibitor in the cell lines expressing mutant or wild type EGFR. Figure S3. In vivo effects of the GNS compounds. Figure S4. In vivo anti-tumor effects of the GNS compounds. Figure S5. In vivo anti-tumor effects of the GNS compounds in intracranial EGFR-mutant lung cancer. Figure S6. In vivo anti-tumor effects of the indicated EGFR inhibitors in intracranial EGFR-mutant lung cancer.

The clinical utility of approved EGFR small-molecule kinase inhibitors is plagued both by toxicity against wild-type EGFR and by metastatic progression in the central nervous system, a disease sanctuary site. Here, we report the discovery and preclinical efficacy of GNS-1486 and GNS-1481, two novel small-molecule EGFR kinase inhibitors that are selective for T790M-mutant isoforms of EGFR. Both agents were effective in multiple mouse xenograft models of human lung adenocarcinoma (T790M-positive or -negative), exhibiting less activity against wild-type EGFR than existing approved EGFR kinase inhibitors (including osimertinib). In addition, GNS-1486 showed superior potency against intracranial metastasis of EGFR-mutant lung adenocarcinoma. Our results offer a preclinical proof of concept for new EGFR kinase inhibitors with the potential to improve therapeutic index and efficacy against brain metastases in patients. Cancer Res; 77(5); 1200–11. ©2017 AACR.

Due to the acceleration of information and communication technology in the Fourth Industrial Revolution, artificial intelligence technology has had a large impact on politics, economy, culture, and art industries. During the COVID-19 pandemic, as face-to-face activities declined and video conferencing expanded, a new platform, called the metaverse, appeared. The metaverse is a digital communication technology, that become known to the public through games and entertainment, and it is gradually expanding. The metaverse also provides new education methods. For high value-added industries, the metaverse is used to foster skilled experts in a shortened training period. In the education field, the metaverse platform is being used in convergence with information technology (IT) companies. Examples include the entrance ceremony, graduation ceremony, job fair, museum experiential learning, and graduation work exhibition of the virtual world through the metaverse. In this paper, college students’ experience of using the metaverse was studied to determine their awareness of and experience with the metaverse. Based on the potential of the metaverse platform, the MZ (Millennials and Generation Z) generation’s intention to use the metaverse, and their experience with it, it is expected that this study will aid the development of the metaverse.

Extracellular vesicles (EVs) have shown great potential in disease diagnosis and treatment; however, their clinical applications remain challenging due to their unsatisfactory long-term stability and the lack of effective delivery strategies. In this study, we prepared human adipose stem cell-derived EV (hASC-EV)-loaded hyaluronic acid dissolving microneedles (EV@MN) to investigate the feasibility of EVs for their clinical applications. The biological activities of the EVs in this formulation were maintained for more than six months under mild storage conditions, especially at temperatures lower than 4 °C. Moreover, the EV@MN enabled precise and convenient intradermal delivery for sustained release of EVs in the dermis layer. Therefore, EV@MN significantly improved the biological functions of hASC-EVs on dermal fibroblasts by promoting syntheses of proteins for the extracellular matrix such as collagen and elastin, enhancing fibroblast proliferation, and regulating the phenotype of fibroblast, compared with other administration methods. This research revealed a possible and feasible formulation for the clinical application of EVs.

In this study, an inkjet bioprinting-based high-throughput screening (HTS) system was designed and applied for the first time to a catecholpyrimidine-based small molecule library to find hit compounds that inhibit c-Jun NH2-terminal kinase1 (JNK1). JNK1 kinase, inactivated MAPKAPK2, and specific fluorescent peptides along with bioink were printed on parchment paper under optimized printing conditions that did not allow rapid evaporation of printed media based on Triton-X and glycerol. Subsequently, different small compounds were printed and tested against JNK1 kinase to evaluate their degree of phosphorylation inhibition. After printing and incubation, fluorescence intensities from the phosphorylated/nonphosphorylated peptide were acquired for the % phosphorylation analysis. The IM50 (inhibitory mole 50) value was determined as 1.55 × 10–15 mol for the hit compound, 22. Thus, this work demonstrated that inkjet bioprinting-based HTS can potentially be adopted for the drug discovery process using small molecule libraries, and cost-effective HTS can be expected to be established based on its low nano- to picoliter printing volume.

Allogeneic transplantation of mesenchymal stem cell-derived extracellular vesicles (EVs) offers great potential for treating liver fibrosis. However, owing to their intrinsic surface characteristics, bare EVs are non-specifically distributed in the liver tissue after systemic administration, leading to limited therapeutic efficacy. To target activated hepatic stellate cells (HSCs), which are responsible for hepatic fibrogenesis, vitamin A-coupled small EVs (V-EVs) were prepared by incorporating vitamin A derivative into the membrane of bare EVs. No significant differences were found in the particle size and morphology between bare and V-EVs. In addition, surface engineering of EVs did not affect the expression of surface marker proteins (e.g., CD63 and CD9), as demonstrated by flow cytometry. Owing to the surface incorporation of vitamin A, V-EVs were selectively taken up by activated HSCs via receptor-mediated endocytosis. When systemically administered to mice with liver fibrosis, V-EVs effectively targeted activated HSCs in the liver tissue, resulting in reversal of the fibrotic cascade. Consequently, even at a 10-fold lower dose, V-EVs exhibited comparable anti-fibrotic effects to those of bare EVs, substantiating their therapeutic potential for liver fibrosis.

Surface engineering of exosomes enhances the therapeutic efficacy of rheumatoid arthritis by macrophage reprogramming., Despite the remarkable advances in therapeutics for rheumatoid arthritis (RA), a large number of patients still lack effective countermeasures. Recently, the reprogramming of macrophages to an immunoregulatory phenotype has emerged as a promising therapeutic strategy for RA. Here, we report metabolically engineered exosomes that have been surface-modified for the targeted reprogramming of macrophages. Qualified exosomes were readily harvested from metabolically engineered stem cells by tangential flow filtration at a high yield while maintaining their innate immunomodulatory components. When systemically administered into mice with collagen-induced arthritis, these exosomes effectively accumulated in the inflamed joints, inducing a cascade of anti-inflammatory events via macrophage phenotype regulation. The level of therapeutic efficacy obtained with bare exosomes was achievable with the engineered exosomes of 10 times less dose. On the basis of the boosted nature to reprogram the synovial microenvironment, the engineered exosomes display considerable potential to be developed as a next-generation drug for RA.

Amyloid-β (Aβ) is generated by proteolytic processing of amyloid precursor protein (APP) by beta-secretase (BACE-1) and gamma-secretase. Amyloid-β is responsible for the formation of senile plaques in Alzheimer's disease (AD). Consequently, inhibition of β-secretase (BACE-1), a rate-limiting enzyme in the production of Aβ, constitutes an attractive therapeutic approach to the treatment of AD. This paper reports an inkjet printing-based fluorescence assay for high throughput screening of β-secretase inhibitors achieved by employing a BACE-1 FRET substrate (Rh-Glu-Val-Asn-Leu-Asp-Ala-Glu-Phe-Lys-Quencher). This peptide substrate is known to be a readily available and suitable substrate for proteolytic activity, and it has high affinity to BACE-1. The BACE-1 peptide substrate printed on parchment paper was effectively cleaved by BACE-1, which was printed on the same spot. The amount of enzyme and substrate required for this inkjet printing-based BACE-1 assay can be less than 1.4ⅹ103, permitting the evaluation of inhibitor activity with femtomolar potency. The inkjet-printing-based BACE-1 inhibitory assay revealed inhibitory effects of inhibitor IV and STA on BACE-1 with an IM50 of 1.00 × 10−15 mol and 1.01 × 10−14 mol, respectively. These data confirm that both BACE-1 inhibitors (inhibitor IV and STA) actively inhibited the BACE-1 proteolysis of BACE-1 substrate on parchment paper. It important to note that the number of mole of BACE-1-substrate and enzyme utilized in the printing-based enzymatic assay are 1.4ⅹ103 smaller than the amount used in the conventional well-plate assay. The inkjet printing-based inhibitory assay constitutes a versatile high throughput technique and the IM50 values of the inhibitors were obtained with satisfactory reproducibility, suggesting that this inkjet-printing BACE-1 inhibitory assay could be quite suitable for the screening of new potential BACE-1 inhibitors for AD.

세계의 대학들은 한편으로 비용 대비 효과성에 대한 질문으로, 또 다른 한편으로 세계적인 경쟁에서 비교우위를 선점하는 데 있어 대학이 수행하는 역할의 중요성에 대한 인식으로 전례 없는 책무성 압력을 경험하고 있다. 이러한 책무성 요구에 대 한 능동적인 반응으로 대학 랭킹이나 교수 1인당 논문 수와 같은 고등교육 조직의 효과성을 나타내주는 다양한 준거들이 개발되고 사용되고 있다. 과거에 활용되었던 방법이나 비판을 받았던 관점에서 벗어나 조직 효과성 측정을 위해 최근 새롭게 조명되는 제3의 접근법은 대학에서의 경험이 학습 성과에 어떠한 영향을 미치는지를 직접적으로 분석하려는 시도이다. 이와 관련하여 지금까지 축적 된 경험에도 불구하고 명확한 한계점은 체계적인 학습경험 데이터의 수집이 이루어 지고 있지 않거나, 데이터가 축적되고 있더라도 유용한 정보로 가공되고 있지 않다 는 점이다. 기존의 연구의 한계를 극복하고자 본 연구는 위계적 군집분석과 회귀분석을 통해 고등교육 공동체에 두 가지 정책적 시사점을 제공하였다. 첫째는 학습경험 측정치 들이 단순한 숫자가 아닌 적절한 맥락 속에서 해석되기 위한 방법론적 측면의 도움 이며, 둘째는 그러한 학습경험 관련 데이터를 평소 각 대학에서 체계적으로 수집하 고 있는 행정관련 데이터와 병합하여 분석함으로써 고등교육 현안에 대한 정책적 시사점을 제공할 수 있다는 점이다. 본 연구는 학생의 학습경험과 학습 성과에 근 거한 대학의 효과성 평가를 위한 방향을 제시하였다는 점에서 그 의의를 찾을 수 있다.

A peptide probe-based MMP-9 inhibitory assay was developed to assess anticancer activity of compounds on parchment paper using FRET-based inkjet printing. The fluorescent peptide containing 5-FAM (donor) and QXL520 (acceptor) was used as a substrate with the sequence of Pro-Leu-Gly-Cys-His-Ala-Arg-Lys specific to MMP-9. The suitability of the peptide as a substrate of MMP-9 was confirmed by HPLC. When both MMP-9 and the peptide were printed on the identical reaction spot of parchment paper, the peptide was cleaved and fragmented by MMP-9. The fluorescence was then recovered by 5-FAM and detected at Ex/Em = 490 ± 20 nm/520 ± 20 nm. Based on the successful performance of FRET on the parchment paper without any surface treatment, the assay platform was applied to test compounds, and a successful anticancer assay could be accomplished. This methodology holds notable strengths in versatile applications to the MMP enzyme family, as well as control of ejection volume, reduction of consumed enzyme amount, and rapid reaction time. Hence, we suggest the FRET peptide-based assay using inkjet printing as a viable replacement for sensitive and high-throughput evaluation of anticancer compounds.

A graphene oxide (GO)-based cost-effective, automatted strip test has developed for screening of inhibitors of endonuclease EcoRV. The method involves the use of GO and a DNA substrate for EcoRV that contains both an ssDNA region for binding of GO and a fluorescein amidite (FAM)-labelled dsDNA. All the components were inkjet printed on a piece of parchment paper. The ssDNA region binds to the surface of GO and anchors so that the fluorescence of FAM is quenched. The parchment paper strip is then incubated with a sample containing EcoRV which causes enzymatic hydrolysis, and dsDNA was separated from the GO. As a result, green fluorescence is generated at the reaction spot. Enzyme activity can be measured in the presence and absence of aurintricarboxy acid acting as an EcoRV inhibitor. This method excels by its need for 2-3 orders less reagents compared to the standard well plate assay. Thus, it is an efficient platform for GO-based screening of EcoRV enzyme inhibitors. Graphical abstract A graphene oxide (GO)-based endonuclease EcoRV inhibition FRET assay using inkjet printing was developed. Printing of GO along with assay reagents has a beneficial effect on the enzymatic reaction on paper. This method was successfully applied to evaluate EcoRV inhibitor activity.

In this study, we report the photo-induced electron transfer (PET) on parchment paper using riboflavin as a photo inducer and ultraviolet lamp (362 nm) as the light source. To this end, a conventional inkjet printer equipped with 4 cartridges was used. Parchment paper was found to be a favorable substrate due to its insignificant self-absorption while assisting efficient sample interaction. Upon UV-irradiation, riboflavin generated superoxide anion radical (O2−·) and it was available to interact with superoxide dismutase present on the same spot. A decrease in NBT formazan in the reaction spot indicates increased O2−· scavenging activity of molecule. It was estimated that the well-plate based-colorimetric method used 12.5 μM (1.25 × 10−9 mole) of riboflavin and 0.25 mM (2.50 × 10−8 mole) of NBT to react with different superoxide dismutase or drug concentrations, while the printing technique consumed 3.19 × 10−13 mole of NBT and 2.98 × 10−13 mole of riboflavin to react with gradient superoxide dismutase or drug concentration. In contrast to the conventional well plate method, inkjet printing-based molecular assay provides automatic delivery in the nanoliter range with precise time, which ensures four-to five-order lower reagent consumption. The inkjet printing-based quantitative measurement specifies the amount of a particular drug/enzyme printed on a surface. Therefore, applicability of inkjet printing technique conjoined radical scavenging assay will be more competent to determine the radical scavenging potential of natural plant products. In addition, this inkjet printing approach offers easy, fast, cost-effective, and less time-consuming method to determine PET reaction on paper.

For the first time, a paper-based fluorescence resonance energy transfer (FRET) determination with cyclic AMP (cAMP)-specific phosphodiesterase 4B (PDE4B) inhibitory assay using an inkjet-printing technique is proposed. Non-fabricated parchment paper is found to constitute a unique substrate to measure fluorescent energy transfer, due to its insignificant self-absorption, and enables efficient sample interaction. Here, we report the responsive FRET signals generated on paper, upon sequentially printing reaction components on parchment paper using a conventional inkjet printer equipped with four cartridges. After printing, the energy emitted by Eu chelate was transferred by FRET to ULight molecule on paper, detected at 665 nm. In the absence of free cAMP, a maximum FRET signal was achieved on paper, while a decrease in FRET signals was recorded when free cAMP produced by PDE4B inhibitors compete with Eu-cAMP, binding with ULight-mAb. The IM50 value was determined as 2.46 × 10−13 mole for roliparm and 1.86 × 10−13 mole for roflumilast, to effectively inhibit PDE4B activity. Inkjet printing-based FRET signal determination utilizes components that are less than the femtomole range, which was four-orders less than the standard assay method. The methodology reported here constitutes an innovative approach towards the determination of FRET signals generated on paper.

The clinical utility of approved EGFR small-molecule kinase inhibitors is plagued both by toxicity against wild-type EGFR and by metastatic progression in the central nervous system, a disease sanctuary site. Here, we report the discovery and preclinical efficacy of GNS-1486 and GNS-1481, two novel small-molecule EGFR kinase inhibitors that are selective for T790M-mutant isoforms of EGFR. Both agents were effective in multiple mouse xenograft models of human lung adenocarcinoma (T790M-positive or -negative), exhibiting less activity against wild-type EGFR than existing approved EGFR kinase inhibitors (including osimertinib). In addition, GNS-1486 showed superior potency against intracranial metastasis of EGFR-mutant lung adenocarcinoma. Our results offer a preclinical proof of concept for new EGFR kinase inhibitors with the potential to improve therapeutic index and efficacy against brain metastases in patients. Cancer Res; 77(5); 1200–11. ©2017 AACR.

Background The association between obesity and response to neoadjuvant chemotherapy in breast cancer patients is not clear. We evaluated the impact of obesity on response to neoadjuvant chemotherapy in patients with operable breast cancer. Methods From May 2008 to December 2010, 104 patients were diagnosed with invasive breast cancer at Korea University Anam Hospital and received neoadjuvant chemotherapy before surgery. Patients were classified into those of normal (BMI of 18.5 to Results Median age was 45 years. Mean BMI was 24.8 kg/m2; 53.8% had a normal BMI, 35.6% overweight, and 10.6% of patients was obese. BMI did not show a significant association with ER status, PR status, HER-2 status, lymph node involvement and neoadjuvant chemotherapy regimen. In univariate analysis, overweight and obese patients were significantly less likely to have a pCR compared with normal weight patients (odds ratio [OR] = 0.300; 95% CI, 0.115 to 0.784; p = 0.010). In multivariate analysis, ER negativity was significantly associated with a pCR and pPR to neoadjuvant chemotherapy (OR = 2.987; 95% CI, 1.128 to 7.907; p = 0.028), And there was significant difference in pCR for overweight and obese compared with normal weight patients (OR = 0.304;95% CI, 0.115 to 0.803; p = 0.016). Conclusion This study suggests that higher BMI should be considered to be a factor of worse response to neoadjuvant chemotherapy in patients with operable breast cancer. Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P1-08-22.

An accurate, rapid, and cost-effective methodology for enzyme inhibitor assays is highly needed for large-scale screening to evaluate the efficacy of drugs at the molecular level. For the first time, we have developed an inkjet printing-based enzyme inhibition assay for the assessment of drug activity using a conventional inkjet printer composed of four cartridges. The methodology is based on the determination of the number of moles of the drug on the printed surface. The number of moles was quantified through the volume of substance ejected onto the printed surface. The volume ejected on the reaction spot was determined from the density of reagent ink solution and its weight loss after printing. A xanthine oxidase (XOD) inhibition assay was executed to quantitatively evaluate antioxidant activities of the drug based on the determination of the number of moles of the drug ejected by inkjet printing. The assay components of xanthine, nitro blue tetrazolium (NBT), superoxide dismutase (SOD)/drug, and XOD were printed systematically on A4 paper. A gradient range of the number of moles of SOD/drug printed on A4 paper could be successfully obtained. Because of the effect of enzyme activity inhibition, incrementally reduced NBT formazan colors appeared on the paper in a number-of-moles-dependent manner. The observed inhibitory mole (IM

Aberrant activations of Fms-like tyrosine receptor kinase (FLT) 3 are implicated in the pathogenesis of 20% to 30% of patients with acute myeloid leukemia (AML). G-749 is a novel FLT3 inhibitor that showed potent and sustained inhibition of the FLT3 wild type and mutants including FLT3-ITD, FLT3-D835Y, FLT3-ITD/N676D, and FLT3-ITD/F691L in cellular assays. G-749 retained its inhibitory potency in various drug-resistance milieus such as patient plasma, FLT3 ligand surge, and stromal protection. Furthermore, it displayed potent antileukemic activity in bone marrow blasts from AML patients regardless of FLT3 mutation status, including those with little or only minor responses to AC220 or PKC412. Oral administration of G-749 yielded complete tumor regression and increased life span in animal models. Thus, G-749 appears to be a promising next-generation drug candidate for the treatment of relapsed and refractory AML patients with various FLT3-ITD/FLT3-TKD mutants and further shows the ability to overcome drug resistance.