Chronotype and subjective sleep quality predict white matter integrity in young people with emerging mental disorders.

Protecting brain health is a goal of early intervention. We explored whether sleep quality or chronotype could predict white matter (WM) integrity in emerging mental disorders. Young people (N = 364) accessing early-intervention clinics underwent assessments for chronotype, subjective sleep quality, and diffusion tensor imaging. Using machine learning, we examined whether chronotype or sleep quality (alongside diagnostic and demographic factors) could predict four measures of WM integrity: fractional anisotropy (FA), and radial, axial, and mean diffusivities (RD, AD and MD). We prioritised tracts that showed a univariate association with sleep quality or chronotype and considered predictors identified by ≥80% of machine learning (ML) models as 'important'. The most important predictors of WM integrity were demographics (age, sex and education) and diagnosis (depressive and bipolar disorders). Subjective sleep quality only predicted FA in the perihippocampal cingulum tract, whereas chronotype had limited predictive importance for WM integrity. To further examine links with mood disorders, we conducted a subgroup analysis. In youth with depressive and bipolar disorders, chronotype emerged as an important (often top-ranking) feature, predicting FA in the cingulum (cingulate gyrus), AD in the anterior corona radiata and genu of the corpus callosum, and RD in the corona radiata, anterior corona radiata, and genu of corpus callosum. Subjective quality was not important in this subgroup analysis. In summary, chronotype predicted altered WM integrity in the corona radiata and corpus callosum, whereas subjective sleep quality had a less significant role, suggesting that circadian factors may play a more prominent role in WM integrity in emerging mood disorders.

Entry inhibition of hepatitis B virus using cyclosporin O derivatives with peptoid side chain incorporation.

Hepatitis B virus (HBV) infection is a serious worldwide health problem causing liver cirrhosis and hepatocellular carcinoma. The development of novel therapeutics targeting distinct steps of the HBV life cycle and combination therapy with approved drugs (i.e., nucleot(s)ides, interferon-α) are considered effective strategies for curing HBV. Among these strategies is the development of entry inhibitors that interfere with the host entry step of HBV to prevent viral infection and transmission. Herein, we generated a novel library of cyclosporin O (CsO) derivatives that incorporate peptoid side chains. Twenty-two CsO derivatives were evaluated for membrane permeability, cytotoxicity, and in vitro HBV entry inhibitory activity. The lead compound (i.e., compound 21) showed the greatest potency in the in vitro HBV entry inhibition assay (IC50 = 0.36 ± 0.01 µM) with minimal cytotoxicity. Our peptide-peptoid hybrid CsO scaffold can readily expand chemical diversity and is applicable for screening various targets requiring macrocyclic chemical entities.

Ulmusakidian, a new coumarin glycoside and antifungal phenolic compounds from the root bark of Ulmus davidiana var. japonica.

Bioactivity-driven LC/MS-based phytochemical analysis of the root bark extract of Ulmus davidiana var. japonica led to the isolation of 10 compounds including a new coumarin glycoside derivative, ulmusakidian (1). The structure of the new compound was elucidated using extensive spectroscopic analyses via 1D and 2D NMR spectroscopic data interpretations, HR-ESIMS, and chemical transformation. The isolated compounds 1-10 were tested for their antifungal activity against human fungal pathogens Cryptococcus neoformans and Candida albicans. Compounds 9 and 10 showed antifungal activity against C. neoformans, with the lowest minimal inhibitory concentration (MIC) of 12.5-25.0 µg/mL, whereas none of the compounds showed antifungal activity against C. albicans.

Metformin and Dichloroacetate Suppress Proliferation of Liver Cancer Cells by Inhibiting mTOR Complex 1.

The Warburg effect is important for cancer cell proliferation. This phenomenon can be flexible by interaction between glycolysis and mitochondrial oxidation for energy production. We aimed to investigate the anticancer effects of the pyruvate dehydrogenase kinase inhibitor, dichloroacetate (DCA) and the mitochondrial respiratory complex I inhibitor metformin in liver cancer cells. The anticancer effect of DCA and/or metformin on HepG2, PLC/PRF5 human liver cancer cell lines, MH-134 murine hepatoma cell lines, and primary normal hepatocytes using MTT assay. Inhibition of lactate/ATP production and intracellular reactive oxygen species generation by DCA and metformin was investigated. Inhibition of PI3K/Akt/mTOR complex I was evaluated to see whether it occurred through AMPK signaling. Anticancer effects of a combination treatment of DCA and metformin were evaluated in HCC murine model. The results showed that metformin and DCA effectively induced apoptosis in liver cancer cells. A combination treatment of metformin and DCA did not affect viability of primary normal hepatocytes. Metformin upregulated glycolysis in liver cancer cells, thereby increasing sensitivity to the DCA treatment. Metformin and DCA inhibited mTOR complex I signaling through upregulated AMPK-independent REDD1. In addition, metformin and DCA increased reactive oxygen species levels in liver cancer cells, which induced apoptosis. A combination treatment of metformin and DCA significantly suppressed the tumor growth of liver cancer cells using in vivo xenograft model. Taken together, the combined treatment of metformin and DCA suppressed the growth of liver cancer cells. This strategy may be effective for patients with advanced liver cancer.

Antifungal Phenols from Woodfordia uniflora Collected in Oman.

Woodfordia uniflora is a flowering shrub unique to the Dhofar region of Oman and is used locally as a sedative and remedy for skin infection. However, no study to date has examined the pharmacological properties of this plant, and studies regarding phytochemicals present in W. uniflora are limited. Herein, phytochemical screening of the extract of W. uniflora was performed using LC/MS. Three new phenolic compounds, (±)-woodfordiamycin (1), woodfordic acid (2), and rhamnetin 3-O-(6″-galloyl)-ß-d-glucopyranoside (3), together with 16 known compounds 4-19, were isolated from the antifungal fraction of the extract. The structures of the new compounds were established by NMR and HR-MS data, and their absolute configurations were established using chemical transformations, including Mosher's method, comparison of experimental and calculated electronic circular dichroism data, and gauge-including atomic orbital NMR chemical shift calculations, followed by DP4+ analysis. The isolated compounds (1-19) were tested for antifungal activity against human fungal pathogens Cryptococcus neoformans and Candida albicans. Compounds (±)-1 and 8 showed antifungal activity against C. neoformans, with the lowest minimum inhibitory concentrations of 1.8-1.9 µM, which was ∼10-fold lower than that of the currently available antifungal drug fluconazole, while (±)-1, 8, and 19 showed antifungal activity against C. albicans.

Polyhalogenation of Isoflavonoids by the Termite-Associated Actinomadura sp. RB99.

Based on high-resolution tandem mass spectrometry (HR-MS2) and global natural products social molecular networking (GNPS), we found that plant-derived daidzein and genistein derivatives are polyhalogenated by termite-associated Actinomadura species RB99. MS-guided purification from extracts of bacteria grown under optimized conditions led to the isolation of eight polychlorinated isoflavones, including six unreported derivatives, and seven novel polybrominated derivatives, two of which showed antimicrobial activity.

Two-Step Real-Time Night-Time Fire Detection in an Urban Environment Using Static ELASTIC-YOLOv3 and Temporal Fire-Tube.

While the number of casualties and amount of property damage caused by fires in urban areas are increasing each year, studies on their automatic detection have not maintained pace with the scale of such fire damage. Camera-based fire detection systems have numerous advantages over conventional sensor-based methods, but most research in this area has been limited to daytime use. However, night-time fire detection in urban areas is more difficult to achieve than daytime detection owing to the presence of ambient lighting such as headlights, neon signs, and streetlights. Therefore, in this study, we propose an algorithm that can quickly detect a fire at night in urban areas by reflecting its night-time characteristics. It is termed ELASTIC-YOLOv3 (which is an improvement over the existing YOLOv3) to detect fire candidate areas quickly and accurately, regardless of the size of the fire during the pre-processing stage. To reflect the dynamic characteristics of a night-time flame, N frames are accumulated to create a temporal fire-tube, and a histogram of the optical flow of the flame is extracted from the fire-tube and converted into a bag-of-features (BoF) histogram. The BoF is then applied to a random forest classifier, which achieves a fast classification and high classification performance of the tabular features to verify a fire candidate. Based on a performance comparison against a few other state-of-the-art fire detection methods, the proposed method can increase the fire detection at night compared to deep neural network (DNN)-based methods and achieves a reduced processing time without any loss in accuracy.

Light-Weight Student LSTM for Real-Time Wildfire Smoke Detection.

As the need for wildfire detection increases, research on wildfire smoke detection combining low-cost cameras and deep learning technology is increasing. Camera-based wildfire smoke detection is inexpensive, allowing for a quick detection, and allows a smoke to be checked by the naked eye. However, because a surveillance system must rely only on visual characteristics, it often erroneously detects fog and clouds as smoke. In this study, a combination of a You-Only-Look-Once detector and a long short-term memory (LSTM) classifier is applied to improve the performance of wildfire smoke detection by reflecting on the spatial and temporal characteristics of wildfire smoke. However, because it is necessary to lighten the heavy LSTM model for real-time smoke detection, in this paper, we propose a new method for applying the teacher-student framework to deep LSTM. Through this method, a shallow student LSTM is designed to reduce the number of layers and cells constituting the LSTM model while maintaining the original deep LSTM performance. As the experimental results indicate, our proposed method achieves up to an 8.4-fold decrease in the number of parameters and a faster processing time than the teacher LSTM while maintaining a similar detection performance as deep LSTM using several state-of-the-art methods on a wildfire benchmark dataset.

Evaluation of drug susceptibility test for Efinaconazole compared with conventional antifungal agents.

BACKGROUND: Superficial fungal infections are one of the most common and burdensome skin problems affecting quality of life in patients. Various conventional antifungal agents have been used to treat fungal infections; however, various problems have been reported including drug interaction, drug resistance and low effectiveness. Efinaconazole is a novel antifungal agent, which has proven to be particularly effective against onychomycosis compared with conventional antifungal agents. However, the antifungal efficacy of Efinaconazole for specific strains has not been analysed. OBJECTIVE: We conducted an in-vitro study to measure the antifungal activity of Efinaconazole against strains of Trichophyton rubrum, Trichophyton mentagrophytes and Candida albicans compared with widely-used antifungal drugs. METHODS: We obtained strains of T. rubrum, T. mentagrophytes and C. albicans isolated from patients with onychomycosis and tinea pedis. The minimal inhibitory concentration (MIC) for various strains of fungal species was evaluated for the antifungal susceptibility test. RESULTS: Efinaconazole showed a low MIC against almost strains of dermatophytes and C albicans and also presented low resistance, indicating high potency of efinaconazole for treatment of superficial fungal infections. CONCLUSION: Efinaconazole could be a comparable alternative to replace existing conventional agents.

Loss of amphiregulin reduces myoepithelial cell coverage of mammary ducts and alters breast tumor growth.

BACKGROUND: Amphiregulin (AREG), a ligand of the epidermal growth factor receptor, is not only essential for proper mammary ductal development, but also associated with breast cancer proliferation and growth. In the absence of AREG, mammary ductal growth is stunted and fails to expand. Furthermore, suppression of AREG expression in estrogen receptor-positive breast tumor cells inhibits in-vitro and in-vivo growth. METHODS: We crossed AREG-null (AREG-/-) mice with the murine luminal B breast cancer model, MMTV-PyMT (PyMT), to generate spontaneous breast tumors that lack AREG (AREG-/- PyMT). We evaluated tumor growth, cytokeratin-8 (K8)-positive luminal cells, cytokeratin-14 (K14)-positive myoepithelial cells, and expression of AREG, Ki67, and PyMT. Primary myoepithelial cells from nontumor-bearing AREG+/+ mice underwent fluorescence-activated cell sorting and were adapted to culture for in-vitro coculture studies with AT-3 cells, a cell line derived from C57Bl/6 PyMT mammary tumors. RESULTS: Intriguingly, PyMT-induced lesions progress more rapidly in AREG-/- mice than in AREG+/+ mice. Quantification of K8+ luminal and K14+ myoepithelial cells in non-PyMT AREG-/- mammary glands showed fewer K14+ cells and a thinner myoepithelial layer. Study of AT-3 cells indicated that coculture with myoepithelial cells or exposure to AREG, epidermal growth factor, or basic fibroblast growth factor can suppress PyMT expression. Late-stage AREG-/- PyMT tumors are significantly less solid in structure, with more areas of papillary and cystic growth. Papillary areas appear to be both less proliferative and less necrotic. In The Cancer Genome Atlas database, luminal-B invasive papillary carcinomas have lower AREG expression than luminal B invasive ductal carcinomas. CONCLUSIONS: Our study has revealed a previously unknown role of AREG in myoepithelial cell development and PyMT expression. AREG expression is essential for proper myoepithelial coverage of mammary ducts. Both AREG and myoepithelial cells can suppress PyMT expression. We find that lower AREG expression is associated with invasive papillary breast cancer in both the MMTV-PyMT model and human breast cancer.

Whole genome sequencing analysis of the cutaneous pathogenic yeast Malassezia restricta and identification of the major lipase expressed on the scalp of patients with dandruff.

Malassezia species are opportunistic pathogenic fungi that are frequently associated with seborrhoeic dermatitis, including dandruff. Most Malassezia species are lipid dependent, a property that is compensated by breaking down host sebum into fatty acids by lipases. In this study, we aimed to sequence and analyse the whole genome of Malassezia restricta KCTC 27527, a clinical isolate from a Korean patient with severe dandruff, to search for lipase orthologues and identify the lipase that is the most frequently expressed on the scalp of patients with dandruff. The genome of M. restricta KCTC 27527 was sequenced using the Illumina MiSeq and PacBio platforms. Lipase orthologues were identified by comparison with known lipase genes in the genomes of Malassezia globosa and Malassezia sympodialis. The expression of the identified lipase genes was directly evaluated in swab samples from the scalps of 56 patients with dandruff. We found that, among the identified lipase-encoding genes, the gene encoding lipase homolog MRES_03670, named LIP5 in this study, was the most frequently expressed lipase in the swab samples. Our study provides an overview of the genome of a clinical isolate of M. restricta and fundamental information for elucidating the role of lipases during fungus-host interaction.

The lysine biosynthetic enzyme Lys4 influences iron metabolism, mitochondrial function and virulence in Cryptococcus neoformans.

The lysine biosynthesis pathway via α-aminoadipate in fungi is considered an attractive target for antifungal drugs due to its absence in mammalian hosts. The iron-sulfur cluster-containing enzyme homoaconitase converts homocitrate to homoisocitrate in the lysine biosynthetic pathway, and is encoded by LYS4 in the model yeast Saccharomyces cerevisiae. In this study, we identified the ortholog of LYS4 in the human fungal pathogen, Cryptococcus neoformans, and found that LYS4 expression is regulated by iron levels and by the iron-related transcription factors Hap3 and HapX. Deletion of the LYS4 gene resulted in lysine auxotrophy suggesting that Lys4 is essential for lysine biosynthesis. Our study also revealed that lysine uptake was mediated by two amino acid permeases, Aap2 and Aap3, and influenced by nitrogen catabolite repression (NCR). Furthermore, the lys4 mutant showed increased sensitivity to oxidative stress, agents that challenge cell wall/membrane integrity, and azole antifungal drugs. We showed that these phenotypes were due in part to impaired mitochondrial function as a result of LYS4 deletion, which we propose disrupts iron homeostasis in the organelle. The combination of defects are consistent with our observation that the lys4 mutant was attenuated virulence in a mouse inhalation model of cryptococcosis.

Characterisation and Expression Analysis of MrLip1, a Class 3 Family Lipase of Malassezia restricta.

The genus Malassezia is associated with a wide range of skin diseases and is the predominant fungal genus isolated from human skin. Of the 14 Malassezia species identified, M. restricta is the most abundant fungal species found from both healthy and diseased skin. Emerging evidences have suggested that extracellular lipases of Malassezia play a critical role in its survival on the host skin surface. This study aimed to characterise the lipase 1 homologue (MrLip1) in M. restricta and to analyse its expression under different environmental conditions. The full sequence of the gene encoding MrLip1 was determined by rapid amplification of cDNA ends, and it was then heterologously expressed in Pichia pastoris. MrLip1 protein was successfully purified and used for lipase assay and specific antibody generation for use in expression analysis. The optimum pH and temperature for the activity of purified MrLip1 were pH 5.0 and 34 °C respectively. Furthermore, the expression of MrLip1 peaked at a similar pH and temperature, suggesting that the optimal conditions for MrLip1 protein activity and expression are similar to that found on the human skin surface. This study provides data to improve our understanding of the role and characteristics of lipase 1 in M. restricta.

Multivariate analyses for monitoring EDCs and PPCPs in a lake water.

The analysis of endocrine disrupting compounds (EDCs) and pharmaceutical and personal care products (PPCP), present at trace level in surface waters, is often expensive, time-consuming, and complex. Implementing effective monitoring strategies for these compounds is essential to determine the types of analytes, sampling locations, and sampling frequencies. Multivariate analyses were used to investigate the patterns of EDCs and PPCPs in Lake Mead, Nevada, for these purposes. The results of cluster analysis and principal component analysis to identify the patterns among compounds demonstrated that selected pharmaceuticals tended to be present together with each other, whereas hormones did not show patterns with other compounds. The results of cluster analysis and discriminant analysis to investigate the spatial variation of EDCs and PPCPs eliminated redundant sampling locations, verifying the current selection of sampling locations in Lake Mead. The results of autocorrelation provided optimal sampling frequencies for EDCs and PPCPs, suggesting either monthly or quarterly monitoring of these compounds in Lake Mead. The patterns of the compounds could be site specific; depending on weather and hydrological conditions of the water systems, but this study's approaches will facilitate effective assessment and monitoring of EDCs and PPCPs in surface water.

Citrus peel pectin and alginate-based emulgel particles for small intestine-targeted oral delivery of curcumin.

Polysaccharides are a prominent choice in the realm of food-grade oral delivery systems due to their resistance to degradation by digestive enzymes in the oral, gastric, and small intestinal environments, as well as their ease of production, cost-effectiveness, and potential health benefits as prebiotics. Furthermore, their ability to respond to pH-induced dissolution, along with their emulsifying properties, can be strategically employed to achieve precise targeting of lipophilic bioactives to the small intestine. In this study, citrus peel pectin and alginate served as stabilizers for emulgel particles without supplementary emulsifiers or gelling agents. Within this system, pectin functioned as an emulsifier, while alginate acted as a gelling agent, facilitated by Ca2+-induced ionic crosslinking. The synergistic interplay between pectin and alginate efficiently protected curcumin in gastric conditions and controlled dissolution in the small intestine, depending on the pectin/alginate ratio. These controlled phenomena facilitated lipolysis, curcumin release, and ultimately enhanced curcumin bioaccessibility. Furthermore, once the emulgel particle released all the entrapped curcumin in the small intestine, residual polysaccharides underwent facile degradation by pectinase and alginate lyase, yielding fermentable monosaccharides. This confirms the potential of the emulgel particles for use as a prebiotic in the colon. These findings offer significant promise for enhancing the systematic design of food-grade delivery systems that encapsulate lipophilic bioactives, achieving controlled release, enhanced stability, and improved bioaccessibility. Importantly, this system can comprise components that undergo complete digestion, absorption, and utilization in the human body, encompassing materials such as oil, nutraceuticals, and prebiotics, all without presenting health risks.

Integrated dark-fermentation-microbial electrosynthesis for efficient wastewater treatment and bioenergy recovery.

High ammonia concentration in wastewater can hinder methane production rate in anaerobic digestion (AD)-microbial electrosynthesis systems (ADMES). To address this issue, a dual-chamber reactor was fabricated using an anion exchange membrane (AEM) to separate the dark-fermentation (DF) and ADMES process, preventing ammonia migration from the DF chamber to the ADMES chamber. As a result, the DF-ADMES achieved a high methane yield based on chemical oxygen demand (COD) of 0.35 L CH4/gCOD compared to control operation AD (0.23 L CH4/gCOD) and ADMES (0.30 L CH4/gCOD). Additionally, hydrogen could be recovered from the DF chamber which improved the energy efficiency of the DF-ADMES reactor (91.7 %) as compared to control AD (53.4 %) and ADMES (71.9 %). Thus, a dual-chamber DF-ADMES with an AEM separator could be a feasible design for scalable treatment of high nitrogen-containing wastewater and high bioenergy recovery.

Genome-wide identification of histone lysine methyltransferases and their implications in the epigenetic regulation of eggshell formation-related genes in a trematode parasite Clonorchis sinensis.

Epigenetic writers including DNA and histone lysine methyltransferases (DNMT and HKMT, respectively) play an initiative role in the differentiation and development of eukaryotic organisms through the spatiotemporal regulation of functional gene expressions. However, the epigenetic mechanisms have long been suspected in helminth parasites lacking the major DNA methyltransferases DNMT1 and DNMT3a/3b. Very little information on the evolutionary status of the epigenetic tools and their role in regulating chromosomal genes is currently available in the parasitic trematodes. We previously suggested the probable role of a DNMT2-like protein (CsDNMT2) as a genuine epigenetic writer in a trematode parasite Clonorchis sinensis. Here, we analyzed the phylogeny of HKMT subfamily members in the liver fluke and other platyhelminth species. The platyhelminth genomes examined conserved genes for the most of SET domain-containing HKMT and Disruptor of Telomeric Silencing 1 subfamilies, while some genes were expanded specifically in certain platyhelminth genomes. Related to the high gene dosages for HKMT activities covering differential but somewhat overlapping substrate specificities, variously methylated histones were recognized throughout the tissues/organs of C. sinensis adults. The temporal expressions of genes involved in eggshell formation were gradually decreased to their lowest levels proportionally to aging, whereas those of some epigenetic tool genes were re-boosted in the later adult stages of the parasite. Furthermore, these expression levels were significantly affected by treatment with DNMT and HKMT inhibitors. Our data strongly suggest that methylated histones are potent epigenetic markers that modulate the spatiotemporal expressions of C. sinensis genes, especially those involved in sexual reproduction.

Optimization of smoothing parameter for block matching and 3D filtering algorithm in low-dose chest and abdominal computed tomography images.

Computed tomography (CT), known for its exceptionally high accuracy, is associated with a substantial dose of ionizing radiation. Low-dose protocols have been devised to address this issue; however, a reduction in the radiation dose can lead to a deficiency in the number of photons, resulting in quantum noise. Thus, the aim of this study was to optimize the smoothing parameter (σ-value) of the block matching and 3D filtering (BM3D) algorithm to effectively reduce noise in low-dose chest and abdominal CT images. Acquired images were subsequently analyze using quantitative evaluation metrics, including contrast to noise ratio (CNR), coefficient of variation (CV), and naturalness image quality evaluator (NIQE). Quantitative evaluation results demonstrated that the optimal σ-value for CNR, CV, and NIQE were 0.10, 0.11, and 0.09 in low-dose chest CT images respectively, whereas those in abdominal images were 0.12, 0.11, and 0.09, respectively. The average of the optimal σ-values, which produced the most improved results, was 0.10, considering both visual and quantitative evaluations. In conclusion, we demonstrated that the optimized BM3D algorithm with σ-value is effective for noise reduction in low-dose chest and abdominal CT images indicating its feasibility of in the clinical field.

Comparative analysis of genetic testing utilization rates among people with and without disabilities in South Korea from 2016 to 2019, focusing on malignant neoplasms: A national population-based study.

INTRODUCTION: Oncogene testing is widely used to detect or direct cancer treatments. Compared to people without disabilities, people with disabilities in Korea have a lower cancer incidence rate but a fivefold higher cancer mortality rate, implying delayed detection. METHODS: We used an administrative database combining disability status and care utilization to analyze every case of cancer-related genetic testing paid for by the National Health Insurance Services of Korea between 2016 and 2019. We first compared percentages of individuals who had taken a registered genetic test by their disability statuses. We then compared the most frequently utilized tests between individuals with and without disabilities. RESULTS: Korean citizens, 175,000 in total, underwent at least one of the 192 registered cancer-related genetic tests between 2016 and 2019. People with disabilities utilized these genetic tests at higher rates than those without disabilities, regardless of sex or age. Among people aged ≥40 years, lung and colorectal cancer-related tests were most frequently utilized, regardless of disability status. CONCLUSION: Although the cancer-related genetic test uptake rate is higher among people with disabilities than among those without disabilities, it is still possible that information on these tests is not as readily available to people with disabilities. Therefore, it is imperative for the government to actively devise strategies to enhance national cancer screening rates among people with disabilities.

Research Note: Comparative evaluation of pathogenicity in SPF chicken between different subgroups of H5N6 high pathogenicity avian influenza viruses.

Since 2014, periodic outbreaks of high pathogenicity avian influenza (HPAI) caused by clade 2.3.4.4 H5 HPAI virus (HPAIV) have resulted in huge economic losses in the Korean poultry industry. During the winter season of 2016-2017, clade 2.3.4.4e H5N6 HPAIVs classified into 5 subgroups (C1-5) were introduced into South Korea. Interestingly, it was revealed that the subgroup C2 and C4 viruses were predominantly distributed throughout the country, whereas detection of the subgroup C3 viruses was confined in a specific local region. In the present study, we conducted comparative evaluation of the pathogenicity of viruses belonging to subgroups C2 and C3 (H15 and HN1 strains) in specific pathogen-free (SPF) chickens, and further compared them with previously determined pathogenicity of subgroup C4 (ES2 strain) virus. The HN1 strain showed lower viral replication in tissues, less transmissibility, and higher mean chicken lethal dose than the H15 and ES2 strains in SPF chickens. Considering that the HN1 strain has a different NS gene segment from the H15 and ES2 strains, the reassortment of the NS gene segment likely affects their infectivity and transmissibility in chickens. These findings emphasize the importance of monitoring the genetic characteristics and pathogenic features of HPAIVs to effectively control their outbreaks in the field.