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Interferon-gamma (IFN-γ) was found to increase in the synovial fluid of patients with rheumatoid arthritis (RA) and osteoarthritis (OA). However, few studies have been conducted to elucidate the role of IFN-γ in cartilage metabolism and regeneration. In this study, we investigated whether cartilage regeneration is driven by interferon-stimulated gene 15 (ISG15) under the control of IFN-γ. IFN-γ significantly increased ITS-induced chondrogenic differentiation of ATDC5 cells. Knockdown of IFN-γ receptor (IFN-γR) inhibited IFN-γ-induced chondrogenic differentiation and reduced ACAN and Col II expression. In addition, ISG15 expression was highly elevated in response to IFN-γ, whereas its expression was downregulated by knockdown of IFN-γR, indicating that ISG15 is closely related to IFN-γ signaling. Furthermore, chondrogenic differentiation and expression of ACAN and Col II were significantly reduced following knockdown of ISG15 in ATDC5 cells despite the presence of IFN-γ. ISGylation of cellular proteins found in chondrogenic differentiated cells was related to activation of IFN-γ signaling. In addition, ISG15/ISGylation was significantly observed in the regenerated cartilage tissue 7 days after FTCI of young mice compared with sham control. Our findings showed that upregulation of ISG15 and/or ISGylation of cellular proteins may play a critical role in cartilage regeneration through activation of IFN-γ signaling., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Inc.)

For the upcycling of waste polyethylene terephthalate (PET), encompassing both colored and fabric PET materials, we investigated the Ir(triNHC)-catalyzed dehydrogenative coupling of PET and methanol, leading to the production of sodium lactate with good yields. We proposed a sustainable method for isolating lactic acid from the catalytic reaction mixture of sodium lactate and regenerating the base using bipolar membrane electrodialysis (BMED). This isolation method demonstrated high effectiveness, achieving isolation of lactic acid while maintaining economic feasibility at $ 0.10 per kg of lactic acid, and enabling sustainable NaOH regeneration with complete resource circulation. We assessed the recyclability of the catalyst and elucidated the mechanism involving base-mediated depolymerization and catalyst-promoted dehydrogenation, highlighting the importance of triNHC ligands in enhancing catalytic activity., (© 2024 The Author(s). Angewandte Chemie International Edition published by Wiley-VCH GmbH.)

Rhodotypos scandens (Thunb.) Makino is known to have a seed dispersal that is thick and stony (endocarp + seeds) and has potential as a landscaping tree seed. In several Rosaceae species, seeds are covered with a hard endocarp, making the internal seeds water-impermeable and germination difficult. Here, we analyzed the morphoanatomical traits and germination properties of R . scandens seeds. To identify ideal seed propagation conditions, we immersed R . scandens seeds in sulfuric acid for varying durations and subjected them to phytohormone (gibberellic acid A3 and fluridone) and a cold stratification (CS) (5 °C) treatment after endocarp removal (ER). The R . scandens stony seeds did not increase in mass by ≥25.0%. Following ER, the seed mass increased by ≥50.0% with water absorption when compared to the initial dry mass. Seed surfaces showed damage and cracks through scarification after 1 h of immersion in sulfuric acid, failing to germinate. A combination of ER, phytohormone treatment, and CS improved seed germination compared to ER alone (26.0 ± 5.3%). Overall, R . scandens seeds showed a dispersal with a hard endocarp from the parent plant, and a pre-treatment with ER, phytohormones, and CS was required for effective seed propagation.

Ir(triNHC) complexes catalyzed glycerol and alcohol dehydrogenative coupling, yielding diverse α-hydroxy acids. Unlike conventional conditions, Ir(triNHC) facilitated additional C-C bond formation after lactic acid production from glycerol, exhibiting high TOFs. This protocol successfully converted 1,2-propanediol and sorbitol into α-hydroxy acids, highlighting biomass-derived sources' potential as valuable platform chemicals.

Background: Mycobacterium tuberculosis is the causative agent of tuberculosis (TB), and its pathogenicity is associated with its ability to evade the host defense system. The secretory form of the chorismate mutase of M. tuberculosis (TBCM, encoded by Rv1885c) is assumed to play a key role in the pathogenesis of TB; however, the mechanism remains unknown., Methods: A tbcm deletion mutant (B∆tbcm) was generated by targeted gene knockout in BCG to investigate the pathogenic role of TBCM in mice or macrophages. We compared the pathogenesis of B∆tbcm and wild-type BCG in vivo by measuring the bacterial clearance rate and the degree of apoptosis. Promotion of the intrinsic apoptotic pathway was evaluated in infected bone marrow-derived macrophages (BMDMs) by measuring apoptotic cell death, loss of mitochondrial membrane potential and translocation of pore-forming proteins. Immunocytochemistry, western blotting and real-time PCR were also performed to assess the related protein expression levels after infection. Furthermore, these findings were validated by complementation of tbcm in BCG., Results: Deletion of the tbcm gene in BCG leads to reduced pathogenesis in a mouse model, compared to wild type BCG, by promoting apoptotic cell death and bacterial clearance. Based on these findings, we found that intrinsic apoptosis and mitochondrial impairment were promoted in B∆tbcm-infected BMDMs. B∆tbcm down-regulates the expression of Bcl-2, which leads to mitochondrial outer membrane permeabilization (MOMP), culminating in cytochrome c release from mitochondria. Consistent with this, transcriptome profiling also indicated that B∆tbcm infection is more closely related to altered mitochondrial-related gene expression than wild-type BCG infection, suggesting an inhibitory role of TBCM in mitochondrial dysfunction. Moreover, genetic complementation of B∆tbcm (C∆tbcm) restored its capacity to inhibit mitochondria-mediated apoptotic cell death., Conclusions: Our findings demonstrate the contribution of TBCM to bacterial survival, inhibiting intrinsic apoptotic cell death of macrophages as a virulence factor of M. tuberculosis complex (MTBC) strains, which could be a potential target for the development of TB therapy., (© 2023. The Author(s).)

Korean bellflower (Campanula takesimana Nakai) is a rare and perennial herb with medicinal and ornamental values, is endemic to the Ulleung Island of Korea. In this study, we investigated the dormancy-release and germination characteristics of C. takesimana (Campanulaceae) seeds by subjecting them to varying temperatures (5, 10, 15, 20, and 25°C and diurnal/nocturnal temperatures of 15/6, 20/10, and 25/15°C), cold stratification periods (0, 4, 8, or 12 weeks at 5°C), and gibberellic acid (GA3) concentrations (0, 10, 100, or 1,000 mg·L-1 at 15/6°C and 25/15°C) to identify the ideal seed propagation conditions. The seeds were stimulated to germinate (at 25°C, 12-h photoperiod with fluorescent lamps at 40 ± 10 μmol∙m-2∙s-1) after cold stratification. To examine the germination characteristics, the seeds were tested for water imbibition and found to readily absorb water. The seeds exhibited underdeveloped embryos during dispersal, showed final germination of 37.00% ± 4.43 at 25°C and were not influenced by temperature. The seeds subjected to 0, 4, 8, or 12 weeks of cold stratification germinated at a success rate of 22.00% ± 4.76, 87.00% ± 6.80, 79.00% ± 2.52, and 77.00% ± 1.91, respectively. Additionally, the germination characteristics, which were based on final germination, mean germination time, and germination velocity (Timson index), were significantly greater in the seeds pretreated with 1,000 mg·L-1 GA3 at 25/15°C than in seeds pretreated with 0 mg·L-1 GA3. Overall, the seeds broke dormancy with GA3 and short-term cold stratification. Therefore, we concluded that C. takesimana seeds have non-deep, simple, morphophysiological dormancy, and pretreatment with cold stratification and GA3 is required for effective seed propagation., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Kim et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Exploiting the heterologous effects of vaccines is a feasible strategy to combat different pathogens. These effects have been explained by enhanced immune responses of innate immune cells. Mycobacterium paragordonae is a rare nontuberculosis mycobacterium that has temperature-sensitive properties. Although natural killer (NK) cells exhibit heterologous immunity features, the cellular crosstalk between NK cells and dendritic cells (DCs) during live mycobacterial infection has remained elusive. We show that live but not dead M. paragordonae enhances heterologous immunity against unrelated pathogens in NK cells by IFN-β of DCs in both mouse models and primary human immune cells. C-di-GMP from live M. paragordonae acted as a viability-associated pathogen-associated molecular pattern (Vita-PAMP), leading to STING-dependent type I IFN production in DCs via the IRE1α/XBP1s pathway. Also, increased cytosolic 2'3'-cGAMP by cGAS can induce type I IFN response in DCs by live M. paragordonae infection. We found that DC-derived IFN-β plays a pivotal role in NK cell activation by live M. paragordonae infection, showing NK cell-mediated nonspecific protective effects against Candida albicans infection in a mouse model. Our findings indicate that the heterologous effect of live M. paragordonae vaccination is mediated by NK cells based on the crosstalk between DCs and NK cells., Competing Interests: Declaration of competing interest The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 The Author(s). Published by Elsevier Masson SAS.. All rights reserved.)

Bacterial leaf blight of carrots caused by Xanthomonas hortorum pv. carotae (Xhc) is an important worldwide seed-borne disease. In 2012 and 2013, symptoms similar to bacterial leaf blight were found in carrot farms in Jeju Island, Korea. The phenotypic characteristics of the Korean isolation strains were similar to the type strain of Xhc. Pathogenicity showed symptoms on the 14th day after inoculation on carrot plants. Identification by genetic method was multi-position sequencing of the isolated strain JJ2001 was performed using four genes (danK, gyrB, fyuA, and rpoD). The isolated strain was confirmed to be most similar to Xhc M081. Furthermore, in order to analyze the genetic characteristics of the isolated strain, whole genome analysis was performed through the next-generation sequencing method. The draft genome size of JJ2001 is 5,443,372 bp, which contains 63.57% of G + C and has 4,547 open reading frames. Specifically, the classification of pathovar can be confirmed to be similar to that of the host lineage. Plant pathogenic factors and determinants of the majority of the secretion system are conserved in strain JJ2001. This genetic information enables detailed comparative analysis in the pathovar stage of pathogenic bacteria. Furthermore, these findings provide basic data for the distribution and diagnosis of Xanthomonas hortorum pv. carotae, a major plant pathogen that infects carrots in Korea.

Fire blight is one of the destructive plant diseases caused by Erwinia amylovora and causes enormous economic losses worldwide. Fire blight was initially reported in apples, pears, and Chinese quince (Park et al. 2016; Myung et al. 2016a, 2016b) in Korea, but recent studies have reported new hosts such as apricot (Lee et al. 2021) and mountain ash (Lim et al, 2023). These reports indicate that fire blight is likely to disperse to new hosts in Korea. During the nationwide survey in June 2021, we observed typical symptoms of blossom blight and shoot blight on a Chinese hawthorn ( Crataegus pinnatifida Bunge) just near an orchard (37°09'21.7"N, 127°35'02.6"E) in Icheon, Gyeonggi Province, where fire blight of Asian pear occurred. For identifying its causal agent, bacterial isolates were recovered after incubating at 28 ℃ for 24 hours on tryptic soy agar (TSA) medium (BD Difco, USA) from blighted leaves and shoots that were surface sterilized with 70% alcohol for 30 sec and homogenized in 500 µl of 10mM MgCl 2 . Pure cultures of white to mucoid colonies were grown on mannitol glutamate yeast extract (MGY) medium, a semi-selective medium for E. amylovora (Shrestha et al, 2003). Two isolates produced 1.5 kb amplicon through colony PCR using amsB primers (Bereswill et al. 1995). Two strains (CPFB26 and CPFB27) from the Chinese hawthorn produced amplicons identical to that from the TS3128 strain of E. amylovora , isolated from the pear tree and identified in 2016 (Park et al. 2016). For the partial 16s rRNA sequences, the total DNA of these two strains was extracted using the Wizard DNA prep kit (Promega, USA), and PCR was performed using fD1 (5'-AGAGTTTGATCCTGGCTCAG-3') and Rp2 (5'-ACGGCTACCTTGTTACGACTT-3') primer sets and further sequenced (Weisburg et al. 1991). These sequences belonged to the E. amylovora clade and were identified as E. amylovora in phylogenetic analysis (GenBank accession no. OP753569 and OP753570). Based on BLASTN analysis, CPFB26 and CPFB27 showed 99.78% similarity to the sequences of the E. amylovora strains TS3128, CFBP 1430, and ATCC 49946. To confirm pathogenicity of the isolates, 10 ㎕ bacterial suspensions (1.5 ⅹ 10 8 CFU/ml) was injected through the veins of the upper 2nd leaf of 3-month-old clone of apple rootstock ( Malus domestica cv. M29) and incubated for six days at 28 ℃ in a chamber with 12 hours of light per day. Petioles and stems turned red hue, and the shoots finally blighted. To complete Koch's postulates, colonies were recovered on TSA medium from the inoculated apple rootstocks and verified through colony PCR for the amsB and A/B primer set (Powney et al. 2011). Hawthorn has been reported as an epidemiologically important alternate host plant of fire blight (van der Zwet et al. 2012). This study is the first to report fire blight caused by E. amylovora in Chinese hawthorn in Korea. Because Chinese hawthorn is natively distributed in Korea and is widely used as a landscaping tree (Jang et al. 2006), the findings of this study suggest that early monitoring could prevent the spread of fire blight through natural hosts.

Introduction: Skin vaccination using dissolving microneedle patch (MNP) technology for transdermal delivery is a promising vaccine delivery strategy to overcome the limitations of the existing vaccine administration strategies using syringes. To improve the traditional microneedle mold fabrication technique, we introduced droplet extension (DEN) to reduce drug loss. Tuberculosis remains a major public health problem worldwide, and BCG revaccination had failed to increase the protective efficacy against tuberculosis. We developed an MNP with live Mycobacterium paragordonae (Mpg) (Mpg-MNP) as a candidate of tuberculosis booster vaccine in a heterologous prime-boost strategy to increase the BCG vaccine efficacy., Materials and Methods: The MNPs were fabricated by the DEN method on a polyvinyl alcohol mask film and hydrocolloid-adhesive sheet with microneedles composed of a mixture of mycobacteria and hyaluronic acid. We assessed the transdermal delivery efficiency by comparing the activation of the dermal immune system with that of subcutaneous injection. A BCG prime Mpg-MNP boost regimen was administered to a mouse model to evaluate the protective efficacy against M. tuberculosis ., Results: We demonstrated the successful transdermal delivery achieved by Mpg-MNP compared with that observed with BCG-MNP or subcutaneous vaccination via an increased abundance of MHCII-expressing Langerin+ cells within the dermis that could migrate into draining lymph nodes to induce T-cell activation. In a BCG prime-boost regimen, Mpg-MNP was more protective than BCG-only immunization or BCG-MNP boost, resulting in a lower bacterial burden in the lungs of mice infected with virulent M. tuberculosis . Mpg-MNP-boosted mice showed higher serum levels of IgG than BCG-MNP-boosted mice. Furthermore, Ag85B-specific T-cells were activated after BCG priming and Mpg-MNP boost, indicating increased production of Th1-related cytokines in response to M. tuberculosis challenge, which is correlated with enhanced protective efficacy., Discussion: The MNP fabricated by the DEN method maintained the viability of Mpg and achieved effective release in the dermis. Our data demonstrate a potential application of Mpg-MNP as a booster vaccine to enhance the efficacy of BCG vaccination against M. tuberculosis . This study produced the first MNP loaded with nontuberculous mycobacteria (NTM) to be used as a heterologous booster vaccine with verified protective efficacy against M. tuberculosis., Competing Interests: Authors M-SL, JS, JJ and DJ were employed by Raphas Co., Ltd. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Lee, Seo, Lee, Kim, Kim, Lee, Oh, Shin, Jin, Jeong and Kim.)

Fire blight, caused by the bacterial pathogen Erwinia amylovora , is a highly destructive disease of apple and pear. Because the apple tree gets systemically infected with E. amylovora and eventually dies, E. amylovora is a considerably important pathogen in the orchard that requires long-term management. In addition, it is crucial to prevent the spread of the pathogen by expeditious diagnosis. In this study, via comparative approaches to the genome sequences of the strains of various Erwinia spp., we designed specific primers targeting a hypothetical gene that is single copy and located in the chromosomal DNA of E. amylovora . This primer set specifically amplified the DNA of E. amylovora but no other bacteria, including E. pyrifoliae , Pectobacterium spp., Pantoea spp., and Dickeya chrysanthemi . Furthermore, the SYBR Green-based real-time PCR using the primer set allowed accurate estimation of the population of E. amylovora . Developing a rapid and accurate diagnostic method using the novel primer set enables effective defense against pathogen spread through continuous monitoring and quick response.

Study Objectives: This study investigated the altered neural function involved in emotional interference and its role in linking sleep disturbance and depressive/anxiety symptoms in rotating shift workers., Methods: Sixty rotating shift workers and 61 controls performed the emotional Stroop task in three blocks (emotional-related, sleep-related, and neutral words) during functional magnetic resonance imaging (fMRI) assessments. Sleep disturbance and depressive/anxiety symptoms were assessed using self-report measures and sleep diaries. Actigraphy was used to assess the sleep and circadian variables. fMRI scans were performed to compare brain activation during the emotional Stroop task. The proposed moderating models were tested using the PROCESS macro in SPSS software., Results: A significant condition effect on reaction time was detected. Regardless of the group, reaction times were longer in the negative emotional word and sleep-related conditions than in the neutral word condition. Whole-brain analysis revealed that rotating shift workers show greater neural activation in the left dorsolateral prefrontal cortex (DLPFC) compared with controls while performing the emotional Stroop task with negative emotional words. Sleep disturbance was more strongly associated with depressive symptoms when activation of the left DLPFC was higher during the emotional Stroop task with negative words., Conclusions: The left DLPFC may play important roles in increased sensitivity to emotional information, possibly due to circadian misalignment, and has moderating effects on the association between sleep disturbance and depressive symptoms in rotating shift workers. These findings will help to identify possible brain regions where interventions can be performed to correct sleep and mood problems in rotating shift workers., (© The Author(s) 2022. Published by Oxford University Press on behalf of Sleep Research Society. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

We investigated differences in brain activity in response to sleep-related pictures between chronic insomnia disorder (CID) patients and good sleepers (GS), and examined whether brain activity moderated the relationship between depressive symptoms and sleep disturbance in CID patients and GS. This study included 43 patients diagnosed with CID, based on the International Classification of Sleep Disorders-3, and 42 GS. The participants kept a sleep diary, underwent nocturnal polysomnography to measure sleep parameters, and completed self-report questionnaires to assess sleep and psychiatric symptoms. They underwent functional magnetic resonance imaging (fMRI) to examine differences in brain activity in response to sleep-related pictures compared to neutral pictures. A moderated moderation analysis was performed to investigate the moderating role of brain responses to sleep-related pictures in the association between depressive symptoms and sleep disturbance. Compared to GS, the brain responses to sleep-related stimuli were significantly lower in CID patients in the right lateral prefrontal cortex (LPFC) and dorsomedial prefrontal cortex (DMPFC). More severe depressive symptoms were significantly associated with longer sleep latency only when LPFC activity was low in CID patients, but not in GS. LPFC hypoactivity in response to sleep-related stimuli in CID patients could moderate the relationship between depression and sleep disturbance., (© 2022. The Author(s).)

Radish ( Raphanus sativus L.) is an important crop grown in Korea primarily for its use in kimchi, and is cultivated on an area of about 23,000ha. In September 2016, radish leaves were observed with yellowing and wilting symptoms in Gangneung (37.55406°N, 128.84871°E) and Jeongseon (37.42895°N, 128.85882°E), Gangwon province, South Korea. Disease incidence was estimated at approximately 10% in two fields, respectively. About 30% of radish plants (cv. Gwangdongyeoreum) with foliar symptoms exhibited vascular discoloration in the roots. Small pieces of discolored root tissue were surface sterilized in 1% NaOCl for 30s, and then rinsed in sterile water. The tissue pieces were placed on water agar and incubated at 25°C for 10 days. Eight isolates were obtained through single spore isolation, and a representative isolate NC16557 (from Gangneung) was selected for identification. After 14 days, colonies on potato dextrose agar (PDA) were 3.7 cm in diameter and becoming dark due to the formation of microsclerotia. Aerial hyphae were smooth-walled, 3 to 4 μm wide. Conidiophores were erect or slanted, verticillately branched or unbranched, and hyaline. Conidia were hyaline, smooth-walled, non-septate, cylindrical to oval, 4.8 to 6.6 × 2.6 to 3.4 μm (mean 5.7 × 2.9 μm, l/w =2.0, n =80). Microsclerotia were immersed in the agar, composed of rounded, brown-pigmented cells and elongate or irregular in shape, 42 to 60 μm diam. Based on morphology, NC16557 isolate was tentatively identified as Verticillium dahliae (Hawksworth and Talboys, 1970; Isaac, 1967). To confirm taxonomic placement, DNA extracted from mycelia of the same isolate was PCR amplified and sequenced targeting internal transcribed spacer (ITS) regions of rDNA, translation elongation factor 1 alpha (TEF), actin (ACT), tryptophan synthase (TS), and glyceraldehyde-3-phosphate dehydrogenase (GPD) genes (Inderbitzin et al., 2011). The sequences were deposited in GenBank with accession numbers MZ723402, and MZ735720-MZ735723, respectively. The sequencing results showed 100% (ITS=479/479, TEF=500/500, ACT=185/185, and TS=295/295) and 99.61% (GPD=257/258) similarity with the sequences of V. dahlia type strain PD322 (LR026889, HQ414624, HQ206921, HQ414909, and HQ414719) by BLAST. Based on the morphology and multigene sequence analysis, the isolate was identified as V. dahliae . Pathogenicity of two isolates (NC16557 and NC16547) was carried out in the greenhouse using ten 10-day-old seedlings (cv. Gwangdongyeoreum) by root-tip cutting and then soaking the roots in a fungal spore suspension of 10 6 conidia mL-1 for 1 hour. Ten seedlings were treated with sterile distilled water as a control. All plants were placed in the greenhouse at 15°C (night)/25°C (day) with natural light. After 6 weeks, all inoculated plants showed vascular discoloration in the roots while control plants remained asymptomatic. However, the above-ground symptoms of inoculated plants, such as yellowing and wilting, were indistinguishable from control plants. V. dahliae was consistently re-isolated from the symptomatic root tissues and the pathogen identity was confirmed by observing morphological characteristics. Verticillium wilt of radish has been reported from China(Yan et al., 2019). In Korea, this is the first report that V. dahliae causes Verticillium wilt of radish, although Dumin et al. (2020) already reported on Verticillium wilt of Chinese cabbage in Gangwon province, the main production area of Chinese cabbage and radish in summer. With these findings in Chinese cabbage and now radish, it will be critical to identify or develop Verticillium-resistant varieties and other management strategies for the stable production of these crops in Korea.

Study Objectives: In the present study, factors associated with sleep perception were identified by comparing clinical characteristics and polysomnographic variables between insomnia patients with negative and positive sleep state misperception (NSSM and PSSM, respectively)., Methods: Self-reported and objective sleep measures were retrospectively collected, including the Pittsburgh Sleep Quality Index (PSQI), Beck Depression Inventory, and a questionnaire on "morning feeling" after nocturnal polysomnography in 150 patients with insomnia. Based on the misperception index (MI), participants were classified into NSSM (MI > 0, n = 115) and PSSM (MI < 0, n = 35) groups., Results: The PSSM group had more N3 sleep on nocturnal polysomnography than the NSSM group ( P = .002). The NSSM group showed a higher PSQI score ( P < .001), longer self-reported sleep-onset latency (SOL) ( P = .001), and a greater SOL discrepancy ( P = .001). Self-reported feelings of tiredness and morning awakenings in the morning were higher in the NSSM group ( P = .029 and P = .038). The MI negatively correlated with a proportion of N3 sleep ( P = .005) and positively correlated with PSQI ( P < .001), morning awakenings ( P = .01), self-reported SOL ( P < .001), and SOL discrepancy ( P < .001) in patients with insomnia. Multiple regression analysis showed that N3 sleep, PSQI, and morning awakenings were significantly associated with MI in patients with insomnia., Conclusions: The proportion of slow-wave sleep and self-reported measures may be associated with perception of sleep in patients with insomnia. Objective and self-reported characteristics of patients with insomnia should be carefully evaluated and managed because they may influence the perception of sleep., Citation: Yoon G, Lee MH, Oh SM, Choi J-W, Yoon SY, Lee YJ. Negative and positive sleep state misperception in patients with insomnia: factors associated with sleep perception. J Clin Sleep Med . 2022;18(7):1789-1795., (© 2022 American Academy of Sleep Medicine.)

Erwinia amylovora and Erwinia pyrifoliae cause fire blight and black-shoot blight, respectively, in apples and pears. E. pyrifoliae is less pathogenic and has a narrower host range than that of E. amylovora. Fire blight and black-shoot blight exhibit similar symptoms, making it difficult to distinguish one bacterial disease from the other. Molecular tools that differentiate fire blight from black-shoot blight could guide in the implementation of appropriate management strategies to control both diseases. In this study, a primer set was developed to detect and distinguish E. amylovora from E. pyrifoliae by conventional polymerase chain reaction (PCR). The primers produced amplicons of different sizes that were specific to each bacterial species. PCR products from E. amylovora and E. pyrifoliae cells at concentrations of 104 cfu/ml and 107 cfu/ml, respectively, were amplified, which demonstrated sufficient primer detection sensitivity. This primer set provides a simple molecular tool to distinguish between two types of bacterial diseases with similar symptoms.

Background: Unnecessary and inappropriate laboratory testing accounts for a significant portion of waste in health care utilization. The aim of this study was to examine the diagnostic value of the anti-nuclear antibody (ANA) test by examining the rate of ANA associated rheumatic disease (AARD) diagnosis among ANA tested and ANA positive subjects and positive predictive value (PPV) of ANA test leading to AARD diagnosis in different ANA titers and different subsets of patients in 5 hospitals affiliated with a university., Methods: We retrospectively extracted data from all subjects who were tested for ANA from year 2010 to 2019. Those who were first evaluated at or referred to rheumatology were further evaluated with extraction of data including ANA titer and ultimate diagnosis. PPVs for ANA test were evaluated after stratification according to clinically relevant key parameters, such as patient age (younger < 65 years vs. older), sex, and requesting department., Results: From 2010 to 2019, A total of 94,153 patients were tested for ANA, of which 13,600 (14.4% of the total) were positive. AARD was diagnosed in only 0.69% among all ANA tested patients and 4.74% among ANA positive patients. The AARD diagnosis rate of ANA positive patients varied widely from 0.1% to 8.7% by requesting department. Using cutoff values above 1:320 yielded PPVs of 15.6 and 7.8% for all AARs and systemic lupus erythematosus. The PPV was significantly higher in young age (< 65 years) and in women, and when it was requested from internal medicine vs other departments., Conclusion: AARD was diagnosed in less than 1% of all ANA tested patients in university-affiliated hospitals. This result shows that careful consideration before ordering the screening ANA is needed to improve the utility of the test for providers and patients and to reduce health costs spurred by unnecessary testing and its consequences., Competing Interests: The authors have no potential conflicts of interest to disclose., (© 2022 The Korean Academy of Medical Sciences.)

Ir(NHC) (NHC, N -heterocyclic carbene)-catalyzed dehydrogenative coupling of sustainable ethylene glycol and various bioalcohols can produce industrially valuable α-hydroxy acids (AHAs). This study is the first to report the sustainable synthesis of higher C n AHAs, in addition to glycolic acid (C 2 AHA) and lactic acid (C 3 AHA). This catalytic system can be recycled to the seventh cycle while maintaining good yields. A reaction mechanism, including facile dehydrogenation of each alcohol and fast cross-coupling of dehydrogenated aldehydes forming products, was proposed based on 18 O- and 2 H-labeling experiments and electron spray ionization-mass spectrometry (ESI-MS) and NMR spectral analyses.

Severe acute respiratory syndrome coronavirus 2 virus (SARS-CoV-2) infection, which causes coronavirus disease 2019 (COVID-19), has led to many cases and deaths worldwide. Therefore, a number of vaccine candidates have been developed to control the COVID-19 pandemic. Of these, to date, 21 vaccines have received emergency approval for human use in at least one country. However, the recent global emergence of SARS-CoV-2 variants has compromised the efficacy of the currently available vaccines. To protect against these variants, the use of vaccines that modulate T cell-mediated immune responses or innate immune cell memory function, termed trained immunity, is needed. The major advantage of a vaccine that uses bacteria or viral systems for the delivery of COVID-19 antigens is the ability to induce both T cell-mediated and humoral immune responses. In addition, such vaccine systems can also exert off-target effects via the vector itself, mediated partly through trained immunity; compared to other vaccine platforms, suggesting that this approach can provide better protection against even vaccine escape variants. This review presents the current status of the development of COVID-19 vaccines based on recombinant viral and bacterial delivery systems. We also discuss the current status of the use of licensed live vaccines for other infections, including BCG, oral polio and MMR vaccines, to prevent COVID-19 infections., (© 2022. The Microbiological Society of Korea.)

Background: This study investigated the effects of anterior insula (AI) activation on the association between stress and sleep disturbance as a neurobiological basis of the trait-like degree of sleep reactivity to stress. Additionally, it examined the effects of AI activity on the association between sleep disturbance and depression., Methods: The participants were 48 adults. To assess AI activation in response to sleep-related stimuli (SS) compared to neutral stimuli (NS), we extracted mean AI parameter estimates for the SS-NS contrast. We examined whether the interaction between life stress and AI activation would predict sleep disturbance and whether the interaction between sleep disturbance and AI activation would predict depression., Results: At higher levels of bilateral AI activation in response to SS, higher levels of stress were associated with greater sleep disturbance (left AI x stress: b = 1.07, SE = 0.44, p < 0.05; right AI x stress: b = 1.05, SE = 0.48, p < 0.05). In addition, at higher levels of right AI activation, higher levels of sleep disturbance were associated with more severe depressive symptoms (right AI x sleep disturbance: b = 2.55, SE = 1.10, p < 0.05)., Limitation: This study assessed sleep quality and depressive symptoms based on self-reported questionnaires., Conclusion: This study revealed moderating effects of AI activation on the association between negative life stress and sleep disturbance. Additionally, AI activation strengthened the association between sleep disturbance and depression. AI activation may reflect a crucial etiological diathesis for insomnia and stress-related disorders., (Copyright © 2021. Published by Elsevier B.V.)

Manual scoring of sleep stages from polysomnography (PSG) records is essential to understand the sleep quality and architecture. Since the PSG requires specialized personnel, a lab environment, and uncomfortable sensors, non-contact sleep staging methods based on machine learning techniques have been investigated over the past years. In this study, we propose an attention-based bidirectional long short-term memory (Attention Bi-LSTM) model for automatic sleep stage scoring using an impulse-radio ultra-wideband (IR-UWB) radar which can remotely detect vital signs. Sixty-five young (30.0 ± 8.6 yrs.) and healthy volunteers underwent nocturnal PSG and IR-UWB radar measurement simultaneously; From 51 recordings, 26 were used for training, 8 for validation, and 17 for testing. Sixteen features including movement-, respiration-, and heart rate variability-related indices were extracted from the raw IR-UWB signals in each 30-s epoch. Sleep stage classification performances of Attention Bi-LSTM model with optimized hyperparameters were evaluated and compared with those of conventional LSTM networks for same test dataset. In the results, we achieved an accuracy of 82.6 ± 6.7% and a Cohen's kappa coefficient of 0.73 ± 0.11 in the classification of wake stage, REM sleep, light (N1+N2) sleep, and deep (N3) sleep which is significantly higher than the conventional LSTM networks (p < 0.01). Moreover, the classification performances were higher than those reported in comparative studies, demonstrating the effectiveness of the attention mechanism coupled with bi-LSTM networks for the sleep staging using cardiorespiratory signals.

This study investigated the kind of seed dormancy and seed germination of Gentiana triflora var. japonica (Kusn.) H. Hara for developing a seed propagation method. The seeds were collected in October 2020 from plants at Mt. Sobaeksan, Korea. In a water imbibition experiment, seed weights increased by >101.9% of their initial masses over 12 h. Effects of incubation temperature (5, 15, 20, 25, 15/6, or 25/15 °C), cold stratification period (5 °C; 0, 4, 8, or 12 weeks), and gibberellic acid (GA 3 ; 0, 10, 100, or 1000 mg∙L -1 ) and potassium nitrate treatment (KNO 3 ; 0, 1000, 2000, or 4000 mg∙L -1 ) on seed germination were investigated to characterize seed dormancy. These seeds exhibited underdeveloped embryos during seed dispersal. The seeds failed to reach the final germination of 15.0% after treatment at 5, 15, 20, 25, 15/6, or 25/15 °C. After cold stratification for 8 weeks, the germination increased dramatically by >90.0% compared to that at 0 weeks. After the GA 3 treatment, the germination reached >80.0% within 5 days. The final germination was 90.0% in the 100 mg∙L -1 GA 3 treatment group. However, the KNO 3 treatment had no effect on seed germination. Therefore, the G . triflora var. japonica seeds exhibited non-deep simple morphophysiological dormancy.

Background: Narcolepsy is marked by pathologic symptoms including excessive daytime drowsiness and lethargy, even with sufficient nocturnal sleep. There are two types of narcolepsy: type 1 (with cataplexy) and type 2 (without cataplexy). Unlike type 1, for which hypocretin is a biomarker, type 2 narcolepsy has no adequate biomarker to identify the causality of narcoleptic phenomenon. Therefore, we aimed to establish new biomarkers for narcolepsy using the body's systemic networks., Method: Thirty participants (15 with type 2 narcolepsy, 15 healthy controls) were included. We used the time delay stability (TDS) method to examine temporal information and determine relationships among multiple signals. We quantified and analyzed the network connectivity of nine biosignals (brainwaves, cardiac and respiratory information, muscle and eye movements) during nocturnal sleep. In particular, we focused on the differences in network connectivity between groups according to sleep stages and investigated whether the differences could be potential biomarkers to classify both groups by using a support vector machine., Result: In rapid eye movement sleep, the narcolepsy group displayed more connections than the control group (narcolepsy connections: 24.47 ± 2.87, control connections: 21.34 ± 3.49; p = 0.022). The differences were observed in movement and cardiac activity. The performance of the classifier based on connectivity differences was a 0.93 for sensitivity, specificity and accuracy, respectively., Conclusion: Network connectivity with the TDS method may be used as a biomarker to identify differences in the systemic networks of patients with narcolepsy type 2 and healthy controls., (Copyright © 2021 The Authors. Published by Elsevier Ltd.. All rights reserved.)

At present, concerns that the recent global emergence of SARS-CoV-2 variants could compromise the current vaccines have been raised, highlighting the urgent demand for new vaccines capable of eliciting T cell-mediated immune responses, as well as B cell-mediated neutralizing antibody production. In this study, we developed a novel recombinant Mycobacterium paragordonae expressing the SARS-CoV-2 receptor-binding domain (RBD) (rMpg-RBD-7) that is capable of eliciting RBD-specific immune responses in vaccinated mice. The potential use of rMpg-RBD-7 as a vaccine for SARS-CoV-2 infections was evaluated in in vivo using mouse models of two different modules, one for single-dose vaccination and the other for two-dose vaccination. In a single-dose vaccination model, we found that rMpg-RBD-7 versus a heat-killed strain could exert an enhanced cell-mediated immune (CMI) response, as well as a humoral immune response capable of neutralizing the RBD and ACE2 interaction. In a two-dose vaccination model, rMpg-RBD-7 in a two-dose vaccination could also exert a stronger CMI and humoral immune response to neutralize SARS-CoV-2 infections in pseudoviral or live virus infection systems, compared to single dose vaccinations of rMpg-RBD or two-dose RBD protein immunization. In conclusion, our data showed that rMpg-RBD-7 can lead to an enhanced CMI response and humoral immune responses in mice vaccinated with both single- or two-dose vaccination, highlighting its feasibility as a novel vaccine candidate for SARS-CoV-2. To the best of our knowledge, this study is the first in which mycobacteria is used as a delivery system for a SARS-CoV-2 vaccine., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Kim, Jeong, Seo, Lee, Shin and Kim.)

Research on species richness patterns and the advanced elevational Rapoport rule (ERR) has been widespread in recent years; however, there is a lack of such research for the temperate mountainous regions in northeast Asia. Here, we collected plant species from the Seorak Mountain in northeast Asia through field surveys. The species were divided into 11 groups according to the life-form types and phytogeography affinities of each species. The ERR was evaluated using Steven's method and by examining the species richness patterns of each group. The species richness patterns revealed a positive multimodal pattern along the elevation gradient, but phytogeography affinities (increasing trend) and life-form analysis (unimodal) exhibited different patterns. The elevation gradients (1,350 m for the mean elevation-range relationships), which are affected by the boundary effect and different life forms, did not consistently support the ERR. However, herbs as well as rare, endemic, and red list species showed consistent support for the ERR, which could be attributed to the influence by phytogeography affinities. Therefore, the results from Seorak Mountain showed that the ERR was not consistent for different plant life forms in the same area; however, phytogeography affinities could support and explain ERR., Competing Interests: The authors declare that they have no conflict of interest., (© 2021 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.)

Background: The influence of the sympathetic nervous system (SNS) on metabolism of bone and cartilage expressing β-adrenergic receptors (AR) was suggested. Here, we investigated whether the SNS functions as a modulator of cartilage metabolism induced by interleukin-1beta (IL-1β)., Methods: Human articular chondrocytes and articular cartilage were collected from patients with osteoarthritis (OA). Chondrocyte monolayer and cartilage explant culture were stimulated with IL-1β. The activity of β-ARs was modulated by an agonist, norepinephrine (NE), and antagonists, including propranolol, atenolol, nebivolol, and nadolol., Results: The levels of β 1 -, β 2 -, and β 3 -AR in OA cartilage and IL-1β-treated chondrocytes were lower than normal cartilage and untreated cells. Treatment of chondrocytes with IL-1β and β-blockers, including propranolol, atenolol, nebivolol, and nadolol, for 6 h significantly upregulated IL-1β-induced expression of MMP-1, -3, and - 13, compared to chondrocytes treated with IL-1β alone, indicating that antagonism of β-AR confers catabolic signals. On the other hand, NE antagonized IL-1β-induced catabolic response. In addition, NE significantly inhibited IL-1β-induced release of glycosaminoglycan (GAG) from cartilage explant culture. In addition, β-AR activity significantly affected IL-1β-stimulated phosphorylation of JNK and ERK. These results indicate that β-AR signal is associated with cartilage metabolism., Conclusions: Our findings showed that β-ARs is a regulator of cartilage catabolism induced with IL-1β., (© 2021. The Author(s).)

Despite the plant microbiota plays an important role in plant health, little is known about the potential interactions of the flower microbiota with pathogens. In this study, we investigated the microbial community of apple blossoms when infected with Erwinia amylovora. The long-read sequencing technology, which significantly increased the genome sequence resolution, thus enabling the characterization of fire blight-induced changes in the flower microbial community. Each sample showed a unique microbial community at the species level. Pantoea agglomerans and P. allii were the most predominant bacteria in healthy flowers, whereas E. amylovora comprised more than 90% of the microbial population in diseased flowers. Furthermore, gene function analysis revealed that glucose and xylose metabolism were enriched in diseased flowers. Overall, our results showed that the microbiome of apple blossoms is rich in specific bacteria, and the nutritional composition of flowers is important for the incidence and spread of bacterial disease.

Sleep is a unique behavioral state that affects body functions and memory. Although previous studies suggested stimulation methods to enhance sleep, a new method is required that is practical for long-term and unconstrained use by people. In this study, we used a novel closed-loop vibration stimulation method that delivers a stimulus in interaction with the intrinsic heart rhythm and examined the effects of stimulation on sleep and memory. Twelve volunteers participated in the experiment and each underwent one adaptation night and two experimental conditions-a stimulation condition (STIM) and a no-stimulation condition (SHAM). The heart rate variability analysis showed a significant increase in the normalized high frequency and the normalized low frequency significantly decreased under the STIM during the slow-wave sleep (SWS) stage. Furthermore, the synchronization ratio between the heartbeat and the stimulus significantly increased under the STIM in the SWS stage. From the electroencephalogram (EEG) spectral analysis, EEG relative powers of slow-wave activity and theta frequency bands showed a significant increase during the STIM in the SWS stage. Additionally, memory retention significantly increased under the STIM compared to the SHAM. These findings suggest that the closed-loop stimulation improves the SWS-stage depth and memory retention, and further provides a new technique for sleep enhancement., (© Sleep Research Society 2020. Published by Oxford University Press on behalf of the Sleep Research Society. All rights reserved. For permissions, please e-mail journals.permissions@oup.com.)

Study Objectives: To assess the seasonality of restless legs syndrome (RLS) using data from the Korean national health insurance database., Methods: We retrospectively reviewed a randomly selected sample representing 3% of the national health insurance claims database in South Korea. From this sample, we obtained the monthly numbers of patients with RLS and diagnoses from 2009 to 2016, along with prescriptions for monthly dopamine agonists and clonazepam for patients with RLS from 2009 to 2013. Total dopamine agonist and clonazepam doses were converted to levodopa-equivalent doses, and the monthly cumulative prescription dose was calculated. Cosinor analysis was used to evaluate the seasonal pattern of each variable., Results: This study included 11,466 patients with RLS and their diagnoses and 4,887 prescriptions for dopamine agonists and clonazepam. There were significant seasonal patterns in the numbers of patients with RLS (P < .001) and diagnoses (P < .001), both of which peaked in August. The magnitude of the greatest difference in the number of patients with RLS between August (highest) and February (lowest) was 29.96% (95% confidence interval, 24.03-100.80), and that of the number of RLS diagnoses was 39.56% (95% confidence interval, 31.24-47.89). The cumulative prescription dose of medication showed no significant seasonality., Conclusions: Our findings suggest that the prevalence of RLS is seasonally affected, with an increase during summer., (© 2021 American Academy of Sleep Medicine.)

We investigated the differential spatial covariance pattern of blood oxygen level-dependent (BOLD) responses to single-task and multitask functional magnetic resonance imaging (fMRI) between patients with psychophysiological insomnia (PI) and healthy controls (HCs), and evaluated features generated by principal component analysis (PCA) for discrimination of PI from HC, compared to features generated from BOLD responses to single-task fMRI using machine learning methods. In 19 patients with PI and 21 HCs, the mean beta value for each region of interest (ROIbval) was calculated with three contrast images (i.e., sleep-related picture, sleep-related sound, and Stroop stimuli). We performed discrimination analysis and compared with features generated from BOLD responses to single-task fMRI. We applied support vector machine analysis with a least absolute shrinkage and selection operator to evaluate five performance metrics: accuracy, recall, precision, specificity, and F2. Principal component features showed the best classification performance in all aspects of metrics compared to BOLD response to single-task fMRI. Bilateral inferior frontal gyrus (orbital), right calcarine cortex, right lingual gyrus, left inferior occipital gyrus, and left inferior temporal gyrus were identified as the most salient areas by feature selection. Our approach showed better performance in discriminating patients with PI from HCs, compared to single-task fMRI.

Tonicity-responsive enhancer-binding protein (TonEBP; nuclear factor of activated T cells 5) is a transcription factor that responds to changes in osmolality. However, recent studies have shown that it also modulates immune responses under inflammatory conditions independently of hyperosmolality. Fibronectin fragments (FN-fs), which are abundant in the synovial fluid of patients with osteoarthritis (OA), induce expression of matrix metalloproteinases (MMPs) via the toll-like receptor-2 (TLR-2) signaling pathway. In this study we examined whether TonEBP is involved in 29-kDa FN-f-induced expression of MMPs. The expression of TonEBP was significantly higher in human osteoarthritis compared with normal cartilage samples. 29-kDa FN-f affected the expression of MMPs 1, 3, and 13 via TonEBP, and expression and nuclear accumulation of TonEBP were induced by activation of the phospholipase C/NF-κB/MAPK signaling pathway and, in particular, modulated by TLR-2. In addition, 29-kDa FN-f induced the expression of osmoregulatory genes, including Tau-T, SMIT, and AR, as well as voltage-dependent calcium channels via the TonEBP/TLR-2 signaling pathway. These results show that 29-kDa FN-f upregulates MMPs in chondrocytes via the TLR-2/TonEBP signaling pathway.

Study Objectives: Idiopathic rapid eye movement sleep behavior disorder (iRBD), characterized by rapid eye movement sleep without atonia (RSWA) and dream-enactment behavior, has been suggested to be a predictor of α-synucleinopathies. Autonomic instability, represented by heart rate variability, is a common characteristic of both iRBD and α-synucleinopathies. Previous studies reported that RSWA was associated with autonomic dysfunction and was a possible predictor of phenoconversion. Therefore, we sought to compare heart rate variability between iRBD and control groups and explore the relationship between heart rate variability and RSWA in patients with iRBD., Methods: Nocturnal polysomnographic data on 47 patients (28 men, 19 women) diagnosed with iRBD based on video-polysomnography and 26 age-matched and sex-matched controls were reviewed. The first 5-minute epoch with a stable electrocardiogram lead II on video-polysomnography was selected from stage N2, wake, and rapid eye movement. For quantification of RSWA, tonic activity was analyzed from the submentalis electromyogram and phasic activity from the submentalis and bilateral anterior tibialis electromyogram channels., Results: Compared to the control group, the iRBD group showed significant reductions in the standard deviation of the R-R intervals, the root mean square of successive R-R interval differences, and high-frequency values. Quantified tonic activity was inversely correlated with normalized low-frequency values and low-frequency/high-frequency ratios and positively correlated with normalized high-frequency values., Conclusions: This study implied decreased cardiac autonomic function in patients with iRBD, which showed parasympathetic predominance. Heart rate variability of the patients with iRBD in this study was associated with quantified tonic RSWA, which was previously reported to be a possible predictor of phenoconversion., (© 2021 American Academy of Sleep Medicine.)

Recently, we reported a 6-mer hepatitis B virus (HBV)-derived peptide, Poly6, that exerts antiviral effects against human immunodeficiency virus type 1 (HIV-1). Here, we explored the immunotherapeutic potential of Poly6 via its administration into dendritic cells (DCs) in a mouse model. Our data revealed that Poly6 treatment led to enhanced production of tumor necrosis factor alpha (TNF-α) and inducible nitric oxide synthase (iNOS)-producing DCs (Tip-DCs) in a type 1 interferon (IFN-I)-dependent manner via the induction of mitochondrial stress. Poly6 treatment in mice implanted with MC38 cells, a murine colon adenocarcinoma line, led to attenuated tumor formation, primarily due to direct cell death induced by Tip-DC mediated nitric oxide (NO) production and indirect killing by Tip-DC mediated cluster of differentiation 8 (CD8) cytotoxic T lymphocyte (CTL) activation via CD40 activation. Moreover, Poly6 treatment demonstrated an enhanced anticancer effect with one of the checkpoint inhibitors, the anti PD-L1 antibody. In conclusion, our data reveal that Poly6 treatment elicits an antitumor immune response in mice, possibly through NO-mediated oncolytic activity via Tip-DC activation and Tip-DC mediated CTL activation. This suggests that Poly6 represents a potential adjuvant for cancer immunotherapy by enhancing the anticancer effects of immune checkpoint inhibitors.

Sleep stage scoring is the first step towards quantitative analysis of sleep using polysomnography (PSG) recordings. However, although PSG is a gold standard method for assessing sleep, it is obtrusive and difficult to apply for long-term sleep monitoring. Further, because human experts manually classify sleep stages, it is time-consuming and exhibits inter-rater variability. Therefore, this article proposes a long short-term memory (LSTM) model for automatic sleep stage scoring using a polyvinylidene fluoride (PVDF) film sensor that can provide unconstrained long-term physiological monitoring. Signals were recorded using a PVDF sensor during PSG. From 60 recordings, 30 were used for training, 10 for validation, and 20 for testing. Sixteen parameters, including movement, respiration-related, and heart rate variability, were extracted from the recorded signals and then normalized. From the selected LSTM architecture, four sleep stage classification performances were evaluated for a test dataset and the results were compared with those of conventional machine learning methods. According to epoch-by-epoch (30 s) analysis, the classification performance for the four sleep stages had an average accuracy of 73.9% and a Cohen's kappa coefficient of 0.55. When compared with other machine learning methods, the proposed method achieved the highest classification performance. The use of LSTM networks with the PVDF film sensor has potential for facilitating automatic sleep scoring, and it can be applied for long-term sleep monitoring at home.

Purpose: The mean sleep duration of Koreans is 7 hours 49 minutes, which is the lowest among Organization for Economic Cooperation and Development countries. Although vitamin D deficiency and arthritis are considered modifiable factors influencing restorative sleep, mean vitamin D levels are low (17.33 ng/mL) and arthritis is prevalent among Koreans. This study investigated whether vitamin D deficiency and arthritis increase the risk of short sleep duration., Patients and Methods: The current study used data from the Korean National Health and Nutrition Examination Survey (KNHANES) for 46,405 Korean adults. Vitamin D levels were measured using a radioimmunoassay kit. Sleep duration and arthritis history were assessed using a self-reported questionnaire. Sleep duration was categorized as optimal (7 to <9 hours/day), short (<7 hours/day), and long (≥9 hours/day). Vitamin D levels were grouped as vitamin D sufficiency (≥20 ng/mL), insufficiency (10-19.99 ng/mL), and deficiency (<10 ng/mL). Multiple imputation was used to compensate for missing data. Multinomial logistic regression was used to examine the associations of vitamin D and arthritis status with sleep duration. The regressions were adjusted for covariates, and each model was adjusted for vitamin D and arthritis., Results: Among these participants, 23,078 (49.73%) and 3518 (7.58%) had short and long sleep duration, respectively; 26,724 (57.59%) and 4349 (9.37%) exhibited vitamin D insufficiency and deficiency, respectively; and 4709 (10.15%) were arthritis patients. Compared with vitamin D sufficiency, vitamin D insufficiency and deficiency were associated with short sleep duration (vitamin D insufficiency group: OR = 1.07, 95% CI = 1.01-1.13; vitamin D deficiency group: OR = 1.13, 95% CI = 1.03-1.24). The presence of arthritis was associated with short sleep duration (OR = 1.14, 95% CI = 1.06-1.22) compared with its absence. The interaction between vitamin D deficiency and the presence of arthritis was significantly associated with short sleep duration. However, long sleep duration was not significantly associated with vitamin D levels or arthritis status., Conclusion: Low vitamin D levels and the presence of arthritis were associated with an increased risk of short sleep duration. Additionally, there was a positive interaction effect between vitamin D deficiency and arthritis on short sleep duration., Competing Interests: Sun-Young Kim, Mi Hyun Lee, Weon-Jeong Lim, Soo In Kim, and Yu Jin Lee report no conflicts of interest in this work., (© 2020 Kim et al.)

During a survey in May 2020, symptoms of blight were observed on apricot ( Prunus armeniaca cv. undetermined) in an orchard (37°06´01.5″N 127°57´44.9″E) in Chungju, South Korea, where fire blight of apple occurred. Three apricot trees in the apple orchard were heavily diseased and showed severe shoot blight and stem canker symptoms. Bacterial isolates were recovered on King's medium B from leaves and twigs that were surface-sterilized with 70% alcohol. Colonies with uniform mucoid, smooth surfaces were collected. DNA from nine isolates did not yield an amplicon in a PCR assay for detection of Erwinia pyrifoliae using primer set CPS1/CPS2c (Kim et al . 2001). Each isolate was positive in PCR assays for E. amylovora using primer sets A/B (Bereswill et al . 1992) and AJ75/76 (Llop et al . 2000) that target pEA29. Sequencing of the PCR products resulted in 99.9% (929 bp out of 930 bp) and 100% (747 bp out of 747 bp) identity with sequences of E. amylovora FB20 (GeneBank: CP050240), respectively. Amplifications of the partial 16S rRNA (GeneBank: LC557153) and hrpN (GeneBank: LC575997) genes were performed, and the products were sequenced. The primers used to amplify 16S rRNA were 518F: 5'-CCAGCAGCCGCGGTAATACG-3' and 800R: 5'-TACCAGGGTATCTAATCC-3', and those for the hrpN genes were HRPN1: 5'-ATGAGTCTGAATACAAG-3' and HRPN3c: 5'-GCTTGCCAAGTGCCATA-3'. BLAST analyses showed 99.8% (1439 bp out of 1442 bp) and 100% (1136 bp out of 1136 bp) identities, respectively, to the sequences of E. amylovora FB20. The ability of the isolates to induce a hypersensitive reaction on tobacco ( Nicotiana tabacum cv. Xanthi) leaves was also evaluated. Bacterial suspensions (1.5 ⅹ 10 8 CFU) of 2 isolates were injected into tobacco leaves, and after 48 h, both isolates caused a hypersensitive response. To confirm pathogenicity of isolates, 3-mm-deep holes in five immature apricot (cv. Goldcot) and five immature apple (cv. Fuji) fruits were inoculated with 10 µl bacterial suspension (1.5 ⅹ 10 8 CFU/ml). The inoculated fruits were placed in a humid plastic box. After 7 days at 27℃, severe necrosis and bacterial ooze were present at the inoculated sites in three repeated tests. No symptoms were observed on fruits inoculated with sterile water. To complete Koch's postulates, bacteria were reisolated from the inoculated apricot and apple fruits. PCR using the specific primer sets stated above confirmed the identity as E. amylovora . Thus, based on disease symptoms, sequences, and pathogenicity, the bacterium causing blight of apricot was identified as E. amylovora . Natural infections of E. amylovora on apricot trees have been reported in the Czech Republic and Hungary (Korba and Sillerova 2011; Vegh and Palkovics 2013). Fire blight was observed in the Czech Republic on apricot trees near pear seedlings, which are highly susceptible to E. amylovora (Korba and Sillerova 2011). Natural infections of E. amylovora on Japanese plum planted adjacent to an apple orchard with severe fire blight has been reported in the United States (Mohan and Thomson 1996). Moreover, susceptibility to fire blight has been reported for apricot and Japanese plum cultivars (Mohan and Bijman 1999). To our knowledge, this the first report of fire blight of apricot caused by E. amylovora in Korea. This report is important because it provides evidence that apricot may be an overlooked reservoir for E. amylovora , in addition to apple, pear, and other rosaceous plants, in Korea. An intensive survey for additional host plants for the fire blight pathogen will be continued in Korea. This work was supported by a grant from the Agenda program (PJ01530202) of Rural Development Administration, Republic of Korea.

Eukaryotic initiation factor 4E (eIF4E)-binding protein 1 (4E-BP1) binds eIF4E and represses protein translation by displacing it from the mRNA. In this study, we investigated the influence of 4E-BP1 translational apparatus on the regulation of transforming growth factor-beta 1 (TGF-β1)-induced anabolic signaling in chondrocytes. The level of 4E-BP1 expression was significantly higher in human OA cartilage than normal cartilage. TGF-β1 increased total protein synthesis, including aggrecan (ACAN) and collagen type II (Col II), together with activation of Akt/mTOR signaling pathway. mTOR silencing significantly suppressed ACAN and Col II expressions through decreasing TGF-β1-induced phosphorylation of 4E-BP1. On the contrary, 4E-BP1 knockdown promoted total protein synthesis but suppressed Col II and ACAN expressions with decreased expression of Smad2/3 and Smad4 and increased expression of inhibitory Smad6 and Smad7. TGF-β1 suppressed the interaction of 4E-BP1 and eIF4E and subsequently enhanced protein synthesis. Furthermore, 4E-BP1 regulated translation levels of inhibitory Smads, which decreased the accumulation of nuclear Smad2/3 complexes on the promoter of ACAN and Col II genes, subsequently affecting transcription of ACAN and Col II. These results demonstrated that TGF-β1-modulated phosphorylation of 4EBP1 plays a role in the expression of Col II and ACAN through differential alteration of Smad signaling pathway., (© 2020 Federation of American Societies for Experimental Biology.)

Leaching behaviors and mechanisms of commercialized glass wasteforms to sequester low-level solid-wastes were investigated: SG glass for resin waste and DG-2 glass for dry active waste. After ANS 16.1 leaching test, leachabilities of the nuclides, Co, Cs, and Sr, were all lager than 14, which met the requirement of the US-Nuclear Regulatory Commission. Holes of diameters 5-10 μm remained on the surface of the SG and crevices of lengths 10-50 μm were observed on the surface of the DG-2. We analyzed elemental compositions of the SG and the DG-2 with depths. For the SG, Si, Al, Ca, and Mg were accumulated and Na was depleted up to nearly 1.5 μm compared to an internal glass. For the DG-2, concentrations of B, Na, Al, Ca and Sr started to decrease from 2.5 μm even though other minor elements are still remained their concentrations. We suggested leaching mechanisms: alkali elements including H would diffuse through the holes on the SG, while most of the elements including Si and Al would diffuse through the crevices on the DG-2., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The decision as to whether patients should be admitted to a medical intensive care unit (ICU), in the absence of information concerning survival rates or prognostic factors in survival, is often challenging. We analyzed survival trends in relation to hospital discharge and examined patient and hospital characteristics associated with survival following ICU care, using a sample of nationwide claims data in Korea from 2002 through 2013. The Korean government implements a compulsory social insurance program that covers the country's entire population, and the Korean National Health Insurance Service-National Sample Cohort (NHIS-NSC) data from 2002 based on this program were used for this study. The NHIS-NSC is a stratified random sample of 1,025,340 subjects selected from around 46 million Koreans. We evaluated annual survival trends using the Kaplan-Meier test. Analyses of the relationship between survival and patient and hospital characteristics were performed using Cox regression analyses. Employing a multivariate model, variables were selected using the forward selection method to consider the multicollinearity of variables. A total of 32,553 patients admitted to an ICU between 2002 and 2013 were identified among the eligible beneficiaries. The number of patients who had histories of ICU admission steadily increased throughout the study period, and patients older than 80 years constituted a progressively increasing proportion of ICU admissions, from 7.3% in 2002 to 16.9% in 2007 to 23.1% in 2013. The mean number of mechanical equipment items applied consistently increased, while no difference was observed in the trend for overall 1-year survival in patients following ICU treatment across the study period: the 1-year survival rate ranged from 66.7% (year 2003) to 64.2% (year 2010). Advanced age, cancer, renal failure, pneumonia, and influenza were all associated with heightened risk of mortality within 1 year. Our results should prove useful to older patients and their clinicians in their decisions regarding whether to seek ICU care, with the goals of improving the end-of life care and optimizing resource utilization.

The nucleotide-binding and oligomerization domain (NOD) is an innate pattern recognition receptor that recognizes pathogen- and damage-associated molecular patterns. The 29-kDa amino-terminal fibronectin fragment (29-kDa FN-f) is a matrix degradation product found in the synovial fluids of patients with osteoarthritis (OA). We investigated whether NOD2 was involved in 29-kDa FN-f-induced pro-catabolic gene expression in human chondrocytes. The expression of mRNA and protein was measured using quantitative real-time polymerase chain reaction (qrt-PCR) and Western blot analysis. Small interfering RNAs were used for knockdown of NOD2 and toll-like receptor 2 (TLR-2). An immunoprecipitation assay was performed to examine protein interactions. The NOD2 levels in human OA cartilage were much higher than in normal cartilage. NOD1 and NOD2 expression, as well as pro-inflammatory cytokines, including interleukin-1beta (IL-1β) and tumor necrosis factor-alpha (TNF-α), were upregulated by 29-kDa FN-f in human chondrocytes. NOD2 silencing showed that NOD2 was involved in the 29-kDa FN-f-induced expression of TLR-2. Expressions of IL-6, IL-8, matrix metalloproteinase (MMP)-1, -3, and -13 were also suppressed by TLR-2 knockdown. Furthermore, NOD2 and TLR-2 knockdown data demonstrated that both NOD2 and TLR-2 modulated the expressions of their adaptors, receptorinteracting protein 2 (RIP2) and myeloid differentiation 88, in 29-kDa FN-f-treated chondrocytes. 29-kDa FN-f enhanced the interaction of NOD2, RIP2 and transforming growth factor beta-activated kinase 1 (TAK1), an indispensable signaling intermediate in the TLR-2 signaling pathway, and activated nuclear factor-κB (NF-κB), subsequently leading to increased expressions of pro-inflammatory cytokines and cartilagedegrading enzymes. These results demonstrate that 29-kDa FN-f modulated pro-catabolic responses via cross-regulation of NOD2 and TLR-2 signaling pathways. [BMB Reports 2019; 52(6): 373-378].

The cGAS-STING pathway plays an important role in pathogen-induced activation of the innate immune response. The 29-kDa amino-terminal fibronectin fragment (29-kDa FN-f) found predominantly in the synovial fluid of osteoarthritis (OA) patients increases the expression of catabolic factors via the toll-like receptor-2 (TLR-2) signaling pathway. In this study, we investigated whether 29-kDa FN-f induces inflammatory responses via the cyclic GMP-AMP synthase (cGAS)/stimulator of interferon gene (STING) pathway in human primary chondrocytes. The levels of cGAS and STING were elevated in OA cartilage compared with normal cartilage. Long-term treatment of chondrocytes with 29-kDa FN-f activated the cGAS/STING pathway together with the increased level of gamma-H2AX, a marker of DNA breaks. In addition, the expression of pro-inflammatory cytokines, including granulocytemacrophage colony-stimulating factor (GM-CSF/CSF-2), granulocyte colony-stimulating factor (G-CSF/CSF-3), and type I interferon (IFN-α), was increased more than 100-fold in 29-kDa FN-f-treated chondrocytes. However, knockdown of cGAS and STING suppressed 29-kDa FN-f-induced expression of GM-CSF, G-CSF, and IFN-α together with the decreased activation of TANK-binding kinase 1 (TBK1), interferon regulatory factor 3 (IRF3), and inhibitor protein κBα (IκBα). Furthermore, NOD2 or TLR-2 knockdown suppressed the expression of GM-CSF, G-CSF, and IFN-α as well as decreased the activation of the cGAS/STING pathway in 29-kDa FN-f-treated chondrocytes. These data demonstrate that the cGAS/STING/TBK1/IRF3 pathway plays a critical role in 29-kDa FN-f-induced expression of pro-inflammatory cytokines. [BMB Reports 2019; 52(5): 336-341].

Purpose: This study investigated prescriptions for sedative-hypnotics via data obtained from the Health Insurance Review and Assessment (HIRA) service., Methods: Data on sedative-hypnotic prescriptions from the HIRA service of the Republic of Korea were analyzed from 2011 to 2015. We included prescriptions for subjects > 18 years of age from hospitals and community healthcare centers. In addition, subgroup analyses with a subsample restricted to prescriptions from patients with diagnostic codes F510 (nonorganic insomnia) or G470 (insomnia) were performed. After analyzing the number of prescriptions by individual pharmacy items, the prescription codes were grouped as: (1) benzodiazepines; (2) non-benzodiazepines, including zolpidem; (3) antidepressants; and (4) antipsychotics. We calculated the monthly percent change in the number of prescriptions by drug group using Joinpoint regression., Results: Among the sedative-hypnotic groups, benzodiazepines were the most commonly prescribed drugs in Korea during the study period. As a single sedative-hypnotic item, zolpidem was the most frequently prescribed medication for patients with insomnia. Prescriptions for all groups of sedative-hypnotics increased significantly during the study period. When stratified by age group, antipsychotic prescriptions increased significantly by 0.19-0.21% per month among men and women aged 50-59 years and > 70 years. Prescriptions for antidepressants in 30-39-year-old men increased significantly by 0.20%., Conclusions: Benzodiazepine prescriptions as well as those for antipsychotics and antidepressants to treat insomnia increased during 2011-2015 in Korea. Monitoring the use of sedative-hypnotics at the national level is necessary, especially in the elderly population.

Objective: Continuation-maintenance electroconvulsive therapy (C/M-ECT) is used to prevent relapse or recurrence in patients with severe mental illnesses. We aimed to investigate the effect of C/M-ECT on reducing hospital re-admissions in patients with treatment-resistant schizophrenia., Methods: We applied a mirror-image design by retrospectively examining re-hospitalization rates of 18 patients with schizophrenia spectrum disorders. We compared the numbers of psychiatric admissions during the actual period over which C/M-ECT was administered with the same period prior to the beginning of C/M-ECT., Results: The number of psychiatric admissions was reduced significantly during C/M-ECT (0.33±0.77) compared with that of the same period prior to C/M-ECT (2.67±1.33) (Wilcoxon signed rank Z=-3.663; p ＜0.001)., Conclusion: This finding shows that C/M-ECT augmentation could successfully reduce the re-hospitalization rates in patients with treatment-resistant schizophrenia.

Endothelial Per-Arnt-Sim domain protein-1/hypoxia-inducible factor-2α (EPAS-1/ HIF-2α) is a catabolic transcription factor that regulates osteoarthritis (OA)-related cartilage destruction. Here, we examined whether microRNA-365 (miR-365) affects interleukin (IL)-1β-induced expression of catabolic factors in chondrocytes via regulation of HIF-2α. MiR-365 levels were significantly decreased in human OA cartilage relative to normal cartilage. Overexpression of miR-365 significantly suppressed IL-1β-induced expression of HIF-2α in human articular chondrocytes. Pharmacological inhibition of various IL-1β-associated signaling pathways revealed mitogen-activated protein kinase and nuclear factor-κB as the primary pathways driving IL-1β-mediated decreases in miR-365 and subsequent increase in HIF-2α expression. Using a luciferase reporter assay encoding the 3' untranslated region (UTR) of human HIF-2α mRNA, we showed that overexpression of miR-365 significantly suppressed IL-1β-induced up-regulation of HIF-2α. AGO2 RNA-immunoprecipitation (IP) assay demonstrated that miR-365 and HIF-2α mRNA were enriched in the AGO2-IP fraction in miR-365-transfected primary chondrocytes compared to miR-con-transfected cells, indicating that HIF-2α is a target of miR-365. Furthermore, miR-365 overexpression significantly suppressed IL-1β-induced expression of catabolic factors, including cyclooxygenase-2 and matrix metalloproteinase-1, -3 and -13, in chondrocytes. In pellet culture of primary chondrocytes miR-365 prevented IL-1β-stimulated extracellular matrix loss and matrix metalloproteinase-13 expression. MiR-365 regulates IL-1β-stimulated catabolic effects in human chondrocytes by modulating HIF-2α expression.

Development of the functional endodermis of Arabidopsis thaliana roots is controlled, in part, by GRAS transcription factors, namely SHORT-ROOT (SHR), SCARECROW (SCR), and SCARECROW-LIKE 23 (SCL23). Recently, it has been shown that the SHR-SCR-SCL23 regulatory module is also essential for specification of the endodermis (known as the bundle sheath) in leaves. Nevertheless, compared with what is known about the role of the SHR-SCR-SCL23 regulatory network in roots, the molecular interactions of SHR, SCR, and SCL23 are much less understood in shoots. Here, we show that SHR forms protein complexes with SCL23 to regulate transcription of SCL23 in shoots, similar to the regulation mode of SCR expression. Our results indicate that SHR acts as master regulator to directly activate the expression of SCR and SCL23. In the SHR-SCR-SCL23 network, we found a previously uncharacterized negative feedback loop whereby SCL23 modulates SHR levels. Through molecular, genetic, physiological, and morphological analyses, we also reveal that the SHR-SCR-SCL23 module plays a key role in the formation of the endodermis (known as the starch sheath) in hypocotyls. Taken together, our results provide new insights into the regulatory role of the SHR-SCR-SCL23 network in the endodermis development in both roots and shoots., (Copyright © 2016 The Author. Published by Elsevier Inc. All rights reserved.)

Introduction: Fibronectin fragments (FN-fs) are increased in the cartilage of patients with osteoarthritis (OA) and have a potent chondrolytic effect. However, little is known about the cellular receptors and signaling mechanisms that are mediated by FN-fs. We investigated whether the 29-kDa amino-terminal fibronectin fragment (29-kDa FN-f) regulates cartilage catabolism via the Toll-like receptor (TLR)-2 signaling pathway in human chondrocytes., Methods: Small interfering RNA was used to knock down TLR-2 and myeloid differentiation factor 88 (MyD88). TLR-2 was overexpressed in chondrocytes transfected with a TLR-2 expression plasmid. The expression levels of matrix metalloproteinase (MMP)-1, MMP-3, and MMP-13 were analyzed using quantitative real-time reverse transcription polymerase chain reactions, immunoblotting, or enzyme-linked immunosorbent assay. The effect of TLR-2 on 29-kDa FN-f-mediated signaling pathways was investigated by immunoblotting., Results: TLR-2, TLR-3, TLR-4, and TLR-5 mRNA were significantly overexpressed in OA cartilage compared with normal cartilage, whereas no significant difference of TLR-1 mRNA expression was found. 29-kDa FN-f significantly increased TLR-2 expression in human chondrocytes in a dose- and time-dependent manner. Knockdown of TLR-2 or MyD88, the latter a downstream adaptor of TLR-2, significantly inhibited 29-kDa FN-f-induced MMP production at the mRNA and protein levels. Conversely, TLR-2 overexpression led to enhanced MMP production by 29-kDa FN-f. In addition, TLR-2 knockdown apparently inhibited 29-kDa FN-f-mediated activation of phosphorylated nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor, alpha, and p38, but not of c-Jun N-terminal kinase or extracellular signal-regulated kinase. Exposure to synovial fluid (SF) from affected joints of patients with OA elevated MMP-1, MMP-3, and MMP-13 expression markedly in primary chondrocytes without reducing cell viability. However, TLR-2 knockdown in chondrocytes significantly suppressed SF-induced MMP induction., Conclusions: Our data demonstrate that the MyD88-dependent TLR-2 signaling pathway may be responsible for 29-kDa FN-f-mediated cartilage catabolic responses. Our results will enhance understanding of cartilage catabolic mechanisms driven by cartilage degradation products, including FN-f. The modulation of TLR-2 signaling activated by damage-associated molecular patterns, including 29-kDa FN-f, is a potential therapeutic strategy for the prevention of cartilage degradation in OA.

Objective: To evaluate the correlation between the magnetic resonance imaging findings of functional pituitary adenomas and histological distribution of hormone-secreting cells in pituitary gland., Methods: Forty-nine patients with pathologically confirmed functional micro and macro pituitary adenomas were retrospectively reviewed for its location and growth direction., Results: Micro-prolactin, micro-adrenocorticotropic hormone (ACTH), and micro-growth hormone (GH) producing adenomas showed specific location (P-value <.01). Macro-GH and macro-thyroid-stimulating hormone producing adenomas showed specific growth direction (P-value <.05), whereas macro-prolactin and macro-ACTH producing adenomas did not., Conclusion: The functional pituitary microadenomas' location and macroadenomas' growth pattern correlate well with histological distribution of hormone-secreting cells in pituitary gland., (Copyright © 2015 Elsevier Inc. All rights reserved.)

A paleoparasitological study was performed on soil samples obtained at archaeological sites dating to the Baekje period in ancient Korean history. The samples were obtained from Buyeo, the capital area of the Baekje Kingdom, and from others corresponding to provincial counties of the same period. We found Ascaris lumbricoides and Trichuris trichiura eggs in the Buyeo samples but not any in the samples from the provincial counties. Our results suggest that the parasite-egg contamination patterns were closely correlated with population densities, in that the provincial counties of the Kingdom, compared with the capital Buyeo, were very sparsely populated. This is the first comprehensive paleoparasitological report on soil contamination patterns in the capital of the Baekje Kingdom, one of the most highly populated areas in ancient Korean history.

In a paleo-parasitological analysis of soil samples obtained from V-shaped pits dating to the ancient Baekje period in Korean history, we discovered Ascaris lumbricoides, Trichuris trichiura, and Clonorchis sinensis eggs. In light of the samples' seriously contaminated state, the V-shaped pits might have served as toilets, cesspits, or dung heaps. For a long period of time, researchers scouring archaeological sites in Korea have had difficulties locating such structures. In this context then, the present report is unique because similar kind of the ancient ruins must become an ideal resource for successful sampling in our forthcoming paleoparasitological studies.