Comparative study for metabolomics, antioxidant activity, and molecular docking simulation of the newly bred Korean red rice accessions.

This study analyzed the metabolite profiles and antioxidant capacities of two waxy and non-waxy Korean red rice accessions newly bred. Fifteen phenolic compounds were detected in the rice samples. Accession1 had high fatty acids, phytosterols, and vitamin E; accession3 had high vitamin E and phytosterol; and accession4 had a high total flavonoid. The correlation analysis findings from this study validated the positive association between all the metabolites and antioxidant activity. in silico results revealed that protocatechuic acid had a docking score of -9.541, followed by luteolin, quercetin, and caffeic acid, all of which had significant docking scores and a significant number of contacts. Similarly, molecular dynamics simulations showed that phytochemicals had root mean square deviation values of <2.8 Å with Keap 1, indicating better stability. This study provides valuable insights into potential directions for future investigations and improvements in the functional qualities of other colored rice varieties.

: A case report of re-challenge of immune checkpoint inhibitors after immune-related neurological adverse events: Review of literature.

INTRODUCTION: The indications for immune checkpoint inhibitors (ICIs) are expanding for various cancers because of their durable responses and tolerable safety profiles. Immune-related adverse events (irAEs), including neurological adverse events (nAEs), are associated with ICIs therapy. However, there have been few studies on whether re-challenge with ICIs can be clinically acceptable after neurological AE has improved. PATIENT CONCERNS: A 69-year-old woman with recurrent ovarian cancer undergoing palliative chemotherapy was admitted to our hospital with sudden development of diplopia, dizziness, and gait instability. The patient was administered ICI therapy with anti-angiogenic agents for 9 weeks for 3 cycles. DIAGNOSIS: We performed neurological examination, brain imaging, nerve conduction studies, and serology tests. The patient was diagnosed with Guillain-Barré syndrome variant, an immune-mediated polyneuropathy characterized by a triad of ataxia, areflexia, and ophthalmoplegia. INTERVENTION: After prompt discontinuation of pembrolizumab, the patient was taken intravenous methylprednisolone (2 mg/kg) was administered for 5 days, and her symptoms were partially resolved. With the addition of immunoglobulin 0.4 g/kg for 5 days, her symptoms gradually improved. OUTCOMES: The patient's neurological symptoms improved after immunosuppressive therapy, without sequelae. The NCV showed normal nerve conduction. Unfortunately, because there was little evidence for pembrolizumab rechallenge, pembrolizumab therapy was permanently discontinued, and the tumors eventually progressed.

Geographical origin discrimination of Agaricus bisporus produced by the complete medium: A pilot study in South Korea.

The complete medium used for mushroom cultivation is important for reliable crop production. We aimed to identify how the origin of Agaricus bisporus grown in Korea was affected by complete media produced in different geographical regions using stable isotope ratios (SIRs). We found that the SIR features of A. bisporus significantly depended on the complete media origin used; in particular, it appeared the high δ34S in the Chinese complete medium, low δ34S in the Dutch complete medium, and high δ15N in the Korean complete medium (P < 0.05). The support vector machine method appeared better geo-origin classification of A. bisporus by the complete media compared to a linear discriminant analysis. A large-scale study should be conducted to establish a reliable origin identification model for A. bisporus grown in complete media to improve the global mushroom marketplace.

Improved accuracy of geographical origin identification of shiitake grown in sawdust medium: A compound-specific isotope model-based pilot study.

In countries like South Korea and the USA, origin labeling of shiitake grown using imported Chinese-inoculated medium is an issue. Therefore, we evaluated the use of compound-specific isotope analysis (CSIA) for the accurate identification of the geographical origin of shiitake (Korean, Chinese-inoculated medium, and Chinese); Chinese-inoculated medium shiitake were cultivated in Korea using inoculated sawdust medium from China. The CSIA-discriminant model showed an overall accuracy of 100% in the geographical classification of the original set and 96.4% for the cross-validated set. Glutamate and aspartate δ15N values were the most important variables for differentiating shiitake based on their origins. Compared to that observed upon using the bulk stable isotope analysis, the CSIA model was associated with significantly improved predictability of origin identification. Our findings elucidate the importance of isotope signatures in developing a reliable origin labeling method for shiitake cultured on the sawdust medium for the global market.

Long-term isotopic model study for ecofriendly rice (Oryza sativa L.) authentication: Updating a case study in South Korea.

To overcome the lack of consumer trust in ecofriendly products due to low reliability of ecofriendly certification and decreasing areas certified for growing ecofriendly agricultural products, alternative approaches for reliable certification are required. Isotopic-chemometric analysis has potential for determining organic authenticity, but previous studies have struggled to differentiate the authenticities of different rice types. The present study examined 5-year variations in δ13C and δ15N in ecofriendly and conventional rice sold at retail markets in South Korea, while assessing the feasibility of discriminant models for authentication of organic rice. Supporting vector machine analysis showed 4.4-14.6% better overall predictability of rice types than discriminant analysis and was effective in discriminating organic or conventional rice from pesticide-free rice, potentially enabling high-throughput screening to authenticate organic rice at marketplaces. Our findings provide reliable information for authenticating ecofriendly rice, with a potential to improve consumer safety and thus the confidence in organic products.

Targeting monoamine oxidase A for T cell-based cancer immunotherapy.

Monoamine oxidase A (MAO-A) is an enzyme best known for its function in the brain, where it breaks down neurotransmitters and thereby influences mood and behavior. Small-molecule MAO inhibitors (MAOIs) have been developed and are clinically used for treating depression and other neurological disorders. However, the involvement of MAO-A in antitumor immunity has not been reported. Here, we observed induction of the Maoa gene in tumor-infiltrating immune cells. Maoa knockout mice exhibited enhanced antitumor T cell immunity and suppressed tumor growth. MAOI treatment significantly suppressed tumor growth in preclinical mouse syngeneic and human xenograft tumor models in a T cell-dependent manner. Combining MAOI and anti-PD-1 treatments generated synergistic tumor suppression effects. Clinical data correlation studies associated intratumoral MAOA expression with T cell dysfunction and decreased patient survival in a broad range of cancers. We further demonstrated that MAO-A restrains antitumor T cell immunity through controlling intratumoral T cell autocrine serotonin signaling. Together, these data identify MAO-A as an immune checkpoint and support repurposing MAOI antidepressants for cancer immunotherapy.

An origin identification model for labeling of shiitake (Lentinula edodes).

With the increasing globalization of the food trade across countries and continents, reliable identification of the geographical origin of products is critical. In this study, we describe the limitations of the current origin labeling system for non-soil-based agricultural products and suggest alternative strategies for the identification of the geographical origin of such products. An origin identification model based on stable isotope ratio analysis combined with discriminant analysis is used to evaluate the similarities and dissimilarities between domestic and foreign shiitake mushrooms, including Chinese inoculated sawdust blocks and Chinese origin. The results show a classification sensitivity of 92.0%, classification specificity of 91.5%, and overall accuracy of 93.5%. In particular, δ15N was the most important isotope marker for the identification of the origin of shiitake mushrooms. Hence, the current origin labeling system for mushroom species has to be revised to establish fair trade and avoid improper origin labeling in the global shiitake market.

Magnetic Solid-Phase Extraction of Drugs and Pesticides from Human Plasma Using COOH-mMWCNTs.

Carbon nanotubes (CNTs) are useful for extracting chemical compounds due to their properties, such as surface area and the potential for chemical modification. Especially the formation of CNTs with carboxylic acid functional group makes them disperse in water-based samples and have strong interaction forces with cationizable analytes. Based on these features, carboxylic acid functionalized multi-walled CNTs (COOH-MWCNTs) have been used as extraction sorbents. CNT can also be gathered using an external magnet by forming complex with iron oxide (Fe3O4) magnetic nanoparticles (MNPs). In this study, COOH-MWCNTs with MNPs were subjected to magnetic solid-phase extraction (mSPE) in order to extract the targeted substances such as diphenhydramine, doxylamine, tramadol, escitalopram, zolpidem, diphenamid, paclobutrazol, hexaconazole, cyproconazole and mepronil from human plasma samples. The following five factors were optimized: (i) the ratio of COOH-MWCNTs to MNPs as a sorbent from 1:1 to 1:4; (ii) sorbent amount starting from 12.5 to 75%; (iii) sample pH tested pH 2 to pH 10 with 1 N hydrochloride and 1 N sodium hydroxide; (iv) agitating time from 0 to 4 min and (v) elution solvent. Limit of detection of 10 targeted substances in human plasma were in the range of 0.1-0.4 mg/L. The recovery of targeted substances (except diphenamid) in human plasma was 73.06-110.28% for intra-day and 83.00-107.70% for inter-day and the precision (relative standard deviation, %) in human plasma was 0.3-13.3% for intra-day and 2.9-15.6% for inter-day. The method was applied to nine authentic biological samples from overdose patients in the emergency room of Chungnam National University Hospital. The performance of mSPE was compared with the liquid-liquid extraction method using ethyl acetate. The results showed that the newly developed method in this study can be used for screening analysis in forensic and clinical toxicology.

Regional Characterization Study of Fatty Acids and Tocopherol in Organic Milk as a Tool for Potential Geographical Identification.

Reliable geographical identification can protect producers of excellent agroproducts, and also provide reliable purchasing information to satisfy consumers. Therefore, this study aimed to evaluate the regional and monthly variation in fatty acid (FA) and tocopherol (TOC) in organic milk (OM) and develop a geographical discriminant model of OM in Korea. In this study, OM had α-TOC and showed a regional or monthly difference of 3-5%. Moreover, C16:0, C18:1 n9 cis + trans, C18:0, and C14:0 were the predominant FAs in OM, and OM mostly had higher ∑UFA, including nutritionally desirable FAs; but lower ∑SFA among four regions or in April and August (p < 0.001). The model prepared using stepwise discriminant analysis showed a classification accuracy of 100% for original and cross-validated sample sets. Our results have characterized regional and monthly nutritional variations of OM, thereby potentially suggesting the applicability of a reliable Korean geographical identification labeling system using nutrient compositional analysis of OM.

Comprehensive analysis of the microbial communities and operational parameters of two full-scale anaerobic digestion plants treating food waste in South Korea: Seasonal variation and effect of ammonia.

There are about ninety full-scale anaerobic digestion (AD) plants in South Korea that treat food waste (FW); however, the key diff ;erences in the microbial communities in different seasons and the effects of ammonia in AD remain poorly understood. In this study, the seasonal changes in microbial communities associated with operational parameters of two full-scale ADs (C and W plants) treating FW were analyzed. The organic loading rate (OLR) variability had an influence on the seasonal CH4 yield; the W plant had a lower CH4 yield with an unstable AD performance while the C plant had a higher CH4 yield with a stable AD performance. It was mainly due to the substantially different NH4+ concentration; the W plant had a NH4+ concentration nearly 1.6 times higher compared to the C plant. The high NH4+ presence in the W plant led to the dominance of class Clostridia, and methanogenesis was mostly done by hydrogenotrophs (Methanomassiliicoccus luminyensis). Additionally, the members belonging to Clostridia and Bacteroidia were found at both plants in each season (share ≥0.5%) implying their indispensable role during the anaerobic digestion of FW.

Development of Hematopoietic Stem Cell-Engineered Invariant Natural Killer T Cell Therapy for Cancer.

Invariant natural killer T (iNKT) cells are potent immune cells for targeting cancer; however, their clinical application has been hindered by their low numbers in cancer patients. Here, we developed a proof-of-concept for hematopoietic stem cell-engineered iNKT (HSC-iNKT) cell therapy with the potential to provide therapeutic levels of iNKT cells for a patient's lifetime. Using a human HSC engrafted mouse model and a human iNKT TCR gene engineering approach, we demonstrated the efficient and long-term generation of HSC-iNKT cells in vivo. These HSC-iNKT cells closely resembled endogenous human iNKT cells, could deploy multiple mechanisms to attack tumor cells, and effectively suppressed tumor growth in vivo in multiple human tumor xenograft mouse models. Preclinical safety studies showed no toxicity or tumorigenicity of the HSC-iNKT cell therapy. Collectively, these results demonstrated the feasibility, safety, and cancer therapy potential of the proposed HSC-iNKT cell therapy and laid a foundation for future clinical development.

Urine Multi-drug Screening with GC-MS or LC-MS-MS Using SALLE-hybrid PPT/SPE.

To intoxicated patients in the emergency room, toxicological analysis can be considerably helpful for identifying the involved toxicants. In order to develop a urine multi-drug screening (UmDS) method, gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-tandem mass spectrometry (LC-MS-MS) were used to determine targeted and unknown toxicants in urine. A GC-MS method in scan mode was validated for selectivity, limit of detection (LOD) and recovery. An LC-MS-MS multiple reaction monitoring (MRM) method was validated for lower LOD, recovery and matrix effect. The results of the screening analysis were compared with patient medical records to check the reliability of the screen. Urine samples collected from an emergency room were extracted through a combination of salting-out assisted liquid-liquid extraction (SALLE) and hybrid protein precipitation/solid phase extraction (hybrid PPT/SPE) plates and examined by GC-MS and LC-MS-MS. GC-MS analysis was performed as unknown drug screen and LC-MS-MS analysis was conducted as targeted drug screen. After analysis by GC-MS, a library search was conducted using an in-house library established with the automated mass spectral deconvolution and identification system (AMDISTM). LC-MS-MS used Cliquid®2.0 software for data processing and acquisition in MRM mode. An UmDS method by GC-MS and LC-MS-MS was developed by using a SALLE-hybrid PPT/SPE and in-house library. The results of UmDS by GC-MS and LC-MS-MS showed that toxicants could be identified from 185 emergency room patient samples containing unknown toxicants. Zolpidem, acetaminophen and citalopram were the most frequently encountered drugs in emergency room patients. The UmDS analysis developed in this study can be used effectively to detect toxic substances in a short time. Hence, it could be utilized in clinical and forensic toxicology practices.

miR-146a modulates autoreactive Th17 cell differentiation and regulates organ-specific autoimmunity.

Autoreactive CD4 T cells that differentiate into pathogenic Th17 cells can trigger autoimmune diseases. Therefore, investigating the regulatory network that modulates Th17 differentiation may yield important therapeutic insights. miR-146a has emerged as a critical modulator of immune reactions, but its role in regulating autoreactive Th17 cells and organ-specific autoimmunity remains largely unknown. Here, we have reported that miR-146a-deficient mice developed more severe experimental autoimmune encephalomyelitis (EAE), an animal model of human multiple sclerosis (MS). We bred miR-146a-deficient mice with 2D2 T cell receptor-Tg mice to generate 2D2 CD4 T cells that are deficient in miR-146a and specific for myelin oligodendrocyte glycoprotein (MOG), an autoantigen in the EAE model. miR-146a-deficient 2D2 T cells induced more severe EAE and were more prone to differentiate into Th17 cells. Microarray analysis revealed enhancements in IL-6- and IL-21-induced Th17 differentiation pathways in these T cells. Further study showed that miR-146a inhibited the production of autocrine IL-6 and IL-21 in 2D2 T cells, which in turn reduced their Th17 differentiation. Thus, our study identifies miR-146a as an important molecular brake that blocks the autocrine IL-6- and IL-21-induced Th17 differentiation pathways in autoreactive CD4 T cells, highlighting its potential as a therapeutic target for treating autoimmune diseases.

Propagating Humanized BLT Mice for the Study of Human Immunology and Immunotherapy.

The humanized bone marrow-liver-thymus (BLT) mouse model harbors a nearly complete human immune system, therefore providing a powerful tool to study human immunology and immunotherapy. However, its application is greatly limited by the restricted supply of human CD34+ hematopoietic stem cells and fetal thymus tissues that are needed to generate these mice. The restriction is especially significant for the study of human immune systems with special genetic traits, such as certain human leukocyte antigen (HLA) haplotypes or monogene deficiencies. To circumvent this critical limitation, we have developed a method to quickly propagate established BLT mice. Through secondary transfer of bone marrow cells and human thymus implants from BLT mice into NSG (NOD/SCID/IL-2Rγ-/-) recipient mice, we were able to expand one primary BLT mouse into a colony of 4-5 proBLT (propagated BLT) mice in 6-8 weeks. These proBLT mice reconstituted human immune cells, including T cells, at levels comparable to those of their primary BLT donor mouse. They also faithfully inherited the human immune cell genetic traits from their donor BLT mouse, such as the HLA-A2 haplotype that is of special interest for studying HLA-A2-restricted human T cell immunotherapies. Moreover, an EGFP reporter gene engineered into the human immune system was stably passed from BLT to proBLT mice, making proBLT mice suitable for studying human immune cell gene therapy. This method provides an opportunity to overcome a critical hurdle to utilizing the BLT humanized mouse model and enables its more widespread use as a valuable preclinical research tool.

Electronic structure tailoring and selective adsorption mechanism of metal-coated nanotubes.

Effects of various metal coating (Co, Ti, Pd, W, and Ru) on electronic structures of carbon nanotubes are systematically studied by both ab initio calculations and field-emission experiments. The theoretical results indicate that the adsorption of metal atoms leads to substantial changes in the band structures and work functions of nanotubes. In particular, titanium is found to be the most effective coating material for the application of nanotubes to the field emission display, by lowering the work function and increasing the local density of states near the Fermi level. This is confirmed by the field-emission experiments using Ti-coated nanotubes, which shows enhanced emission performances. In addition, it is found that the Ti coating extends the lifetime of the nanotube substantially. Through the thermogravimetric analysis and theoretical modeling, we propose that this is related to the role of metal coating as a protection layer against residual gases such as oxygen, which cause the degradation of nanotubes. The applications of metal-coated nanotubes to other types of electronic devices are also discussed.