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The human leukocyte antigen (HLA) system, which is part of the major histocompatibility complex (MHC) plays a vital role in immune responses by differentiating between itself and foreign cells and antigens. The significant diversity of alleles affects disease susceptibility and immune responses within different populations. Specifically, the HLA-A and HLA-DRB1 alleles are associated with various immune-related diseases, and understanding the frequency and haplotype associations of these alleles is vital for genetic and immunological research. To investigate the distribution of these characteristics in Koreans, we isolated peripheral blood mononuclear cells (PBMCs) from blood samples donated by volunteers at the Seoul Central Blood Bank and performed HLA typing on 790 samples. Our study found that the HLA-A and HLA-DRB1 alleles are widely distributed within the Korean population, with HLA-A*24:02 (21.7%) and HLA-DRB1*09:01 (9.9%) being the most frequent. Significant haplotype associations between specific HLA-A and HLA-DRB1 alleles were identified using the Chi-square test, suggesting that certain genetic combinations may influence disease onset. This insight could contribute to the development of predictive and preventative strategies for various diseases. The unique genetic characteristics of the Korean population highlight the importance of studying the HLA allele and the haplotype distributions in this group as key indicators for understanding disease susceptibility.

The so-called “Corona Blue” decline in mental health after the COVID-19 outbreak. This study examines how COVID-19 risk perception affects mental health via urban space usage in Seoul, Korea, and Shanghai, China. It uniquely explores direct PR-MH and OS-MH effects, along with indirect effects mediated by pandemic-driven changes in urban space utilization. Results reveal that visits to urban destinations and open spaces mediate the relationship between perceived risk and MH, with differing significance levels between the two cities. In Seoul, disruptions in daily life negatively affected mental health through decreased interactions with urban destinations and outdoor spaces, with a statistically significant impact on stress through urban destinations. This suggests that maintaining access to and engagement with these spaces is crucial for mental health during crises. Conversely, Shanghai showed indirect effects predominantly through perceptions of safety, which significantly influenced anxiety levels through urban destinations, but did not manifest similarly through interactions with outdoor spaces. Caution in interpretation is necessary for better generation, but the outcomes of this study underscore the importance of considering cultural, historical, and political contexts in understanding the varying impacts of future pandemic policies on mental health.

IntroductionAryl hydrocarbon receptor (AhR) is a transcription factor that performs various functions upon ligand activation. Several studies have explored the role of AhR expression in tumor progression and immune surveillance. Nevertheless, investigations on the distribution of AhR expression, specifically in cancer or immune cells in the tumor microenvironment (TME), remain limited. Examining the AhR expression and distribution in the TME is crucial for gaining insights into the mechanism of action of AhR-targeting anticancer agents and their potential as biomarkers.MethodsHere, we used multiplexed immunohistochemistry (mIHC) and image cytometry to investigate the AhR expression and distribution in 513 patient samples, of which 292 are patients with one of five solid cancer types. Additionally, we analyzed the nuclear and cytosolic distribution of AhR expression.ResultsOur findings reveal that AhR expression was primarily localized in cancer cells, followed by stromal T cells and macrophages. Furthermore, we observed a positive correlation between the nuclear and cytosolic expression of AhR, indicating that the expression of AhR as a biomarker is independent of its localization. Interestingly, the expression patterns of AhR were categorized into three clusters based on the cancer type, with high AhR expression levels being found in regulatory T cells (Tregs) in non-small cell lung cancer (NSCLC).DiscussionThese findings are anticipated to serve as pivotal evidence for the design of clinical trials and the analysis of the anticancer mechanisms of AhR-targeting therapies.

IntroductionTo understand the immune system within the tumor microenvironment (TME) of non-small cell lung cancer (NSCLC), it is crucial to elucidate the characteristics of molecules associated with T cell activation.MethodsWe conducted an in-depth analysis using single-cell RNA sequencing data obtained from tissue samples of 19 NSCLC patients. T cells were classified based on the Tumor Proportion Score (TPS) within the tumor region, and molecular markers associated with activation and exhaustion were analyzed in T cells from high TPS areas.ResultsNotably, tetraspanins CD81 and CD82, belonging to the tetraspanin protein family, were found to be expressed in activated T cells, particularly in cytotoxic T cells. These tetraspanins showed strong correlations with activation and exhaustion markers. In vitro experiments confirmed increased expression of CD81 and CD82 in IL-2-stimulated T cells. T cells were categorized into CD81highCD82high and CD81lowCD82low groups based on their expression levels, with CD81highCD82high T cells exhibiting elevated activation markers such as CD25 and CD69 compared to CD81lowCD82low T cells. This trend was consistent across CD3+, CD8+, and CD4+ T cell subsets. Moreover, CD81highCD82high T cells, when stimulated with anti-CD3, demonstrated enhanced secretion of cytokines such as IFN-γ, TNF-α, and IL-2, along with an increase in the proportion of memory T cells. Bulk RNA sequencing results after sorting CD81highCD82high and CD81lowCD82low T cells consistently supported the roles of CD81 and CD82. Experiments with overexpressed CD81 and CD82 showed increased cytotoxicity against target cells.DiscussionThese findings highlight the multifaceted roles of CD81 and CD82 in T cell activation, cytokine production, memory subset accumulation, and target cell cytolysis. Therefore, these findings suggest the potential of CD81 and CD82 as promising candidates for co-stimulatory molecules in immune therapeutic strategies for cancer treatment within the intricate TME.

This study evaluates how different feeding systems impact ruminal fermentation, methane production, and microbiota of Hanwoo steers native to Korea. In a replicated 2 × 2 crossover design over 29 days per period, eight Hanwoo steers (507.1 ± 67.4 kg) were fed twice daily using a separate feeding (SF) system comprising separate concentrate mix and forage or total mixed rations (TMR) in a 15:85 ratio. The TMR-feeding group exhibited a considerable neutral detergent fiber digestibility increase than the SF group. However, ruminal fermentation parameters and methane production did not differ between two feeding strategies. In addition, TMR-fed steers expressed elevated Prevotellaceae family, Christensenellaceae R-7 group, and an unidentified Veillonellaceae family genus abundance in their rumen, whereas SF-fed steers were rich in the Rikenellaceae RC9 gut group, Erysipelotrichaceae UCG-004, and Succinivibrio. Through linear regression modeling, positive correlations were observed between the Shannon Diversity Index and the SF group’s dry matter intake and methane production. Although feeding systems do not affect methane production, they can alter ruminal microbes. These results may guide future feeding system investigations or ruminal microbiota manipulations as a methane-mitigation practice examining different feed ingredients.

The growing interest in healthy diets has driven the demand for food ingredients with enhanced health benefits. In this study, we aimed to explore a method to enhance the bioactivity of kale using a home vertical farming appliance. Specifically, we investigated the effects of treating kale with a green tea water extract (GTE; 0.1–0.5 g/L in nutrient solution) for two weeks before harvest during five weeks of kale cultivation. GTE treatment did not negatively affect the key quality attributes, such as yield, semblance, or sensory properties. However, it led to the accumulation of bioactive compounds, epicatechin (EC) and epigallocatechin gallate (EGCG), which are typically absent in kale. In the control group, no catechins were detected, whereas in the GTE-treated group, the concentration of EC and EGCG were as high as 252.11 and 173.26 μg/g, respectively. These findings indicate the successful incorporation of catechins, known for their unique health-promoting properties, into kale. Additionally, GTE treatment enhanced the biosynthesis of glucosinolates, which are key secondary metabolites of kale. The total glucosinolate content increased from 9.56 μmol/g in the control group to 16.81 μmol/g in the GTE-treated group (treated with 0.5 g/L GTE). These findings showed that GTE treatment not only enriched kale with catechins, the primary bioactive compounds in green tea but also increased the levels of glucosinolates. This study, conducted using a home vertical farming appliance, suggests that bioactivity-enhanced kale can be grown domestically, providing consumers with a nutrient-fortified food source.

Natural killer (NK) cells have recently shown renewed promise as therapeutic cells for use in treating hematologic cancer indications. Despite this promise, NK cell manufacturing workflows remain largely manual, open, and disconnected, and depend on feeders, as well as outdated unit operations or processes, often utilizing research-grade reagents. Successful scale-up of NK cells critically depends on the availability and performance of nutrient-rich expansion media and cryopreservation conditions that are conducive to high cell viability and recovery post-thaw. In this paper we used Cytiva hardware and media to expand the NK92 cell line in a model process that is suitable for GMP and clinical manufacturing of NK cells. We tested a range of cryopreservation factors including cooling rate, a range of DMSO-containing and DMSO-free cryoprotectants, ice nucleation, and cell density. Higher post-thaw recovery was seen in cryobags over cryovials cooled in identical conditions, and cooling rates of 1°C/min or 2°C/min optimal for cryopreservation in DMSO-containing and DMSO-free cryoprotectants respectively. Higher cell densities of 5x107 cells/ml gave higher post-thaw viability than those cryopreserved at either 1x106 or 5x106 cells/ml. This enabled us to automate, close and connect unit operations within the workflow while demonstrating superior expansion and cryopreservation of NK92 cells. Cellular outputs and performance were conducive to clinical dosing regimens, serving as a proof-of-concept for future clinical and commercial manufacturing.

Significant improvement in targeted therapy for colorectal cancer (CRC) has occurred over the past few decades since the approval of the EGFR inhibitor cetuximab. However, cetuximab is used only for patients possessing the wild-type oncogene KRAS, NRAS, and BRAF, and even most of these eventually acquire therapeutic resistance, via activation of parallel oncogenic pathways such as RAS-MAPK or PI3K/Akt/mTOR. The two aforementioned pathways also contribute to the development of therapeutic resistance in CRC patients, due to compensatory and feedback mechanisms. Therefore, combination drug therapies (versus monotherapy) targeting these multiple pathways may be necessary for further efficacy against CRC. In this study, we identified PIK3CA mutant (PIK3CA MT) as a determinant of resistance to SMI-4a, a highly selective PIM1 kinase inhibitor, in CRC cell lines. In CRC cell lines, SMI-4a showed its effect only in PIK3CA wild type (PIK3CA WT) cell lines, while PIK3CA MT cells did not respond to SMI-4a in cell death assays. In vivo xenograft and PDX experiments confirmed that PIK3CA MT is responsible for the resistance to SMI-4a. Inhibition of PIK3CA MT by PI3K inhibitors restored SMI-4a sensitivity in PIK3CA MT CRC cell lines. Taken together, these results demonstrate that sensitivity to SMI-4a is determined by the PIK3CA genotype and that co-targeting of PI3K and PIM1 in PIK3CA MT CRC patients could be a promising and novel therapeutic approach for refractory CRC patients.

The development of alcohol hangover relief drinks by adding persimmon vinegar was investigated in this study. This study aimed to develop and investigate the physicochemical characteristics of a hangover relief drink, derived from persimmon vinegar, that may have health benefits. Persimmon vinegar was added at concentrations of 0, 2.5, 5, 7.5, 10 and 12.5%. The higher the concentration of persimmon vinegar, the lower the pH, °Brix and reducing sugar content of the product. In contrast, higher titratable acidity, turbidity and tannin values were obtained with increasing concentrations of persimmon vinegar. The addition of 12.5% persimmon vinegar induced the highest alcohol dehydrogenase and acetaldehyde dehydrogenase activities at 160.91 and 117.14%, respectively. The L value also decreased as persimmon vinegar concentration increased. The addition of persimmon vinegar at high concentrations decreased fructose, glucose and maltose content but increased the sucrose content of the drink. Ca, K and Na were the most abundant minerals in the drink. Some organic acids, such as oxalic, malic, lactic, acetic, citric and succinic acids, were also detected in the developed alcohol hangover relief drink. This study suggests that adding 7.5% of persimmon vinegar improves the physicochemical characteristics, especially the Alcohol dehydrogenase and aldehyde dehydrogenase activities. This finding indicates that this formulated drink with 75% persimmon vinegar may be beneficial against oxidative stress.

Objective Recently, indigenous Korean grass Sasa quelpaertensis Nakai (SQ) has garnered much interest as a roughage source for livestock to mitigate its adverse effects on habitat diversity. Thus, the objective of the present study was to evaluate the ruminal fermentation, palatability, and nutrient digestibility of SQ for Korean native beef cattle (Hanwoo) using in vitro rumen fermentation, in situ rumen degradability, and in vivo feeding trials. Methods Using in vitro tests with rumen fluid as the inoculum for 48 h, ruminal fermentation of SQ was evaluated and compared with that of other roughage sources commonly used in Korea (i.e., rice straw, Timothy hay, and Italian ryegrass [IRG]). Additionally, an in situ trial 96 h was performed using three cannulated Hanwoo steers. Further, an in vivo trial was performed using eight Hanwoo steers to compare the palatability of SQ with rice straw in total mixed ration (TMR) and forage-concentrate separate feeding conditions. Finally, an in vivo digestibility trial of SQ fed as TMR of two particle sizes was performed with four Hanwoo steers. Results In vitro and in situ trials revealed that SQ was comparable or superior to rice straw in terms of the ruminal fermentation characteristics of pH, gas production, total volatile fatty acid content, and effective ruminal dry matter digestibility (DMD), although its fermentability was lower than that of Timothy hay and IRG. In the palatability test, steers showed a greater preference for SQ when given as TMR. The total tract DMD of SQ fed as TMR was 75.9%±1.37%, and it did not differ by particle size. Conclusion The feed value of SQ as a roughage source for Hanwoo steers is comparable or superior to that of rice straw, particularly when provided as TMR.

The impact of fermented concentrate on the growth and rumen health of beef cattle remains an area of emerging research. This study aimed to assess the influence of a fermented concentrate (TRT) compared to a conventional concentrate (CON) on the growth, rumen fermentation characteristics, and microbiota composition in Korean cattle. Using a crossover design, eight cattle were alternately fed TRT and CON diets, with subsequent analysis of feed components, rumen fermentation parameters, and microbial profiles. TRT and CON diets did not differ significantly in their effect on animal growth metrics. However, the TRT diet was associated with reduced digestibility of rapidly degradable carbohydrates and modified rumen fermentation patterns, as evidenced by an elevated pH and increased acetate-to-propionate ratio (p < 0.05). Furthermore, the TRT diet increased the abundance of lactic acid bacteria, Bacillus, and yeast and organic acid levels in the rumen (p < 0.05). Moreover, Lachnospiraceae and Bacteroidales populations in the rumen and fecal Akkermansia abundance increased in the TRT group compared to the CON group. These microbial changes suggest a potential enhancement of the immune system and overall health of the host. Further research on the long-term implications of incorporating fermented concentrate into cattle diets is warranted.

Magnetic Resonance Imaging (MRI) is sensitive to motion caused by patient movement due to the relatively long data acquisition time. This could cause severe degradation of image quality and therefore affect the overall diagnosis. In this paper, we develop an efficient retrospective 2D deep learning method called stacked U-Nets with self-assisted priors to address the problem of rigid motion artifacts in 3D brain MRI. The proposed work exploits the usage of additional knowledge priors from the corrupted images themselves without the need for additional contrast data. The proposed network learns the missed structural details through sharing auxiliary information from the contiguous slices of the same distorted subject. We further design a refinement stacked U-Nets that facilitates preserving the spatial image details and improves the pixel-to-pixel dependency. To perform network training, simulation of MRI motion artifacts is inevitable. The proposed network is optimized by minimizing the loss of structural similarity (SSIM) using the synthesized motion-corrupted images from 83 real motion-free subjects. We present an intensive analysis using various types of image priors: the proposed self-assisted priors and priors from other image contrast of the same subject. The experimental analysis proves the effectiveness and feasibility of our self-assisted priors since it does not require any further data scans. The overall image quality of the motion-corrected images via the proposed motion correction network significantly improves SSIM from 71.66% to 95.03% and declines the mean square error from 99.25 to 29.76. These results indicate the high similarity of the brain's anatomical structure in the corrected images compared to the motion-free data. The motion-corrected results of both the simulated and real motion data showed the potential of the proposed motion correction network to be feasible and applicable in clinical practices.

Background: Coronavirus disease 2019 (COVID-19) is a disease that is characterized by frequent reinfection. However, the factors influencing reinfection remain poorly elucidated, particularly regarding the effect of COVID-19 vaccination on preventing reinfection and its effects on symptomatology and the interval until reinfection. Methods: This retrospective cohort study examined patients with severe acute respiratory syndrome coronavirus reinfection between January 2020 and February 2022. This study included patients aged >17 years who were reinfected at least 90 days between two infections with severe acute respiratory syndrome coronavirus. The main outcome measure was a reduction in symptoms during reinfection, and reinfection interval. Results: Overall, 712 patients (average age: 40.52 ± 16.41 years; 312 males) were included. The reduction rate of symptoms at reinfection than that at first infection was significantly higher in the vaccinated group than in the unvaccinated group (p < 0.001). The average reinfection interval was 265.81 days. The interval between the first and second infection was 63.47 days longer in the vaccinated group than in the unvaccinated group. The interval was also 57.23 days, significantly longer in the asymptomatic group than in the symptomatic group (p < 0.001). Conclusions: Besides its role in preventing severe acute respiratory syndrome coronavirus infection, vaccination reduces the rate of symptomatic reinfection and increases the reinfection interval; thus, it is necessary to be vaccinated even after a previous infection. The findings may inform the decision to avail COVID-19 vaccination.

Background Intrapleural urokinase is one of the most widely used fibrinolytic agents in the treatment of complicated parapneumonic effusion (CPPE). However, little research has been performed on the optimal urokinase dosage. The aim of this study was to evaluate the treatment efficacy of half dose urokinase compared with conventional dose urokinase. Methods We retrospectively enrolled 92 patients with CPPE or empyema who underwent intrapleural urokinase treatment at two tertiary hospitals. Patients received antibiotics, chest tube drainage, and other treatments as part of routine care. The primary outcome was the treatment success rate in the half dose urokinase group (50,000 IU daily for maximal 6 days) and the conventional dose urokinase group (100,000 IU daily). Treatment success was defined as clinical and radiological improvements without surgical treatment or re-admission within one month. Results Forty-four patients received half dose urokinase, whereas 48 patients were treated with conventional dose urokinase. Both groups were relatively well matched at baseline, excluding higher serum white blood cell count and higher empyema prevalence in the half dose urokinase group. The treatment success rate was not different between the two groups (p=0.048). There were no differences in the rate of in-hospital death and surgical treatment, hospitalization duration, and indwelling catheter duration. In the multivariate analysis, urokinase dose was not a predictor of treatment success. Conclusion Half dose intrapleural urokinase is equally effective conventional dose urokinase in treating patients with CPPE or empyema.

Organic solar cells (OSCs) demonstrating high power conversion efficiencies have been mostly fabricated using halogenated solvents, which are highly toxic and harmful to humans and the environment. Recently, non-halogenated solvents have emerged as a potential alternative. However, there has been limited success in attaining an optimal morphology when non-halogenated solvents (typically o-xylene (XY)) were used. To address this issue, we studied the dependence of the photovoltaic properties of all-polymer solar cells (APSCs) on various high-boiling-point non-halogenated additives. We synthesized PTB7-Th and PNDI2HD-T polymers that are soluble in XY and fabricated PTB7-Th:PNDI2HD-T-based APSCs using XY with five additives: 1,2,4-trimethylbenzene (TMB), indane (IN), tetralin (TN), diphenyl ether (DPE), and dibenzyl ether (DBE). The photovoltaic performance was determined in the following order: XY + IN < XY + TMB < XY + DBE ≤ XY only < XY + DPE < XY + TN. Interestingly, all APSCs processed with an XY solvent system had better photovoltaic properties than APSCs processed with chloroform solution containing 1,8-diiodooctane (CF + DIO). The key reasons for these differences were unraveled using transient photovoltage and two-dimensional grazing incidence X-ray diffraction experiments. The charge lifetimes of APSCs based on XY + TN and XY + DPE were the longest, and their long lifetime was strongly associated with the polymer blend film morphology; the polymer domain sizes were in the nanoscale range, and the blend film surfaces were smoother, as the PTB7-Th polymer domains assumed an untangled, evenly distributed, and internetworked morphology. Our results demonstrate that the use of an additive with an optimal boiling point facilitates the development of polymer blends with a favorable morphology and can contribute to the widespread use of eco-friendly APSCs.

A recently developed treatment strategy for lung cancer that combines immune checkpoint inhibitors with chemotherapy has been applied as a standard treatment for lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC), and it has improved the outcomes of chemotherapy. Maintenance treatment with anti-PD-1 antibody (aPD-1) enhances the effect of immunochemical combination therapy and improves therapeutic efficacy, which contributes toward a significant improvement in patient survival rates. The AXL receptor tyrosine kinase (AXL), which is expressed in tumor cells, plays an essential role in the resistance of cancers to chemotherapy and immunotherapy, and stimulates signaling associated with epithelial-mesenchymal transition (EMT) in metastatic cancer. AXL is thus an attractive target for controlling resistance to anti-tumor therapies. In this study, we examined the effect of AXL inhibitors on immune activation and tumor growth in TC1 and C3PQ mouse tumor models, in the context of clinical immunotherapy/chemotherapy and maintenance treatment, using an aPD-1 with/without pemetrexed. To determine the optimal timing for administration of SKI-G-801, an AXL inhibitor, we investigated its anti-tumor effects based on inclusion at the immunochemotherapy and maintenance therapy stages. We also performed flow cytometry-based immune profiling of myeloid cells and lymphoid cells at different points in the treatment schedule, to investigate the immune activation and anti-tumor effects of the AXL inhibitor. The addition of SKI-G-801 to the immune checkpoint inhibitor and chemotherapy stage, as well as the maintenance therapy stage, produced the best anti-tumor results, and significant tumor growth inhibition was observed in both the TC1 and C3PQ models. Both models also exhibited increased proportion of effector memory helper T cells and increased expression of CD86+ macrophages. Especially, regulatory T cells were significantly reduced in the TC1 tumor model and there was an increase in central memory cytotoxic T cell infiltration and an increased proportion of macrophages with high CD80 expression in the C3PQ tumor model. These results suggest increased infiltration of T cells, consistent with previous studies using AXL inhibitors. It is expected that the results from this study will serve as a stepping stone for clinical research to improve the existing standard of care.

Heat stress negatively affects cattle productivity by reducing feed intake. In the present study, we assessed if the rumen microbiome composition of Hanwoo steers was altered by exposure to heat stress. Rumen samples were collected from four Hanwoo steers that were individually housed in climate-controlled chambers with 60% humidity and environmental temperatures of: 1) 15°C (0-day group), 2) 35°C for 3 days (3-day group), and 3) 35°C for 6 days (6-day group). The total community DNA of samples was extracted, and 997,843 bacterial and 1,508,770 archaeal sequences were analyzed using next-generation sequencing. Assessment of the relative abundances revealed 15 major phyla of which Bacteroidetes was found to be the most dominant. After 3 days of heat stress exposure there were no significant changes in the rumen microbiome composition, except for a decrease in the Planctomycetes. However, after 6 days of heat stress exposure, we found that the relative abundance of fibrolytic Ruminococcaceae had decreased while that of lactate-producing Lactobacillaceae and amylolytic Prevotella and Ruminobacter had increased. The normal rumen microbiome of Hanwoo cattle was shown to be disrupted after 6 days of heat stress, which led to the decrease in fibrolytic bacteria that are sensitive to low pH and the increase in both lactate-producing and amylolytic bacteria. We have demonstrated that the microbiome composition of the rumen is affected by acute heat stress. Our findings may contribute to the development of different feeding strategies to restore heat stress-induced disruption of the rumen microbiome.

Abstract Objectives AXL‐mediated activation of aberrant tyrosine kinase drives various oncogenic processes and facilitates an immunosuppressive microenvironment. We evaluated the anti‐tumor and anti‐metastatic activities of SKI‐G‐801, a small‐molecule inhibitor of AXL, alone and in combination with anti‐PD‐1 therapy. Methods In vitro pAXL inhibition by SKI‐G‐801 was performed in both human and mouse cancer cell lines. Immunocompetent mouse models of tumor were established to measure anti‐metastatic potential of SKI‐G‐801. Furthermore, SKI‐G‐801, anti‐PD‐1 or their combination was administered as an adjuvant or neoadjuvant in the 4T1 tumor model to assess their potential for clinical application. Results SKI‐G‐801 robustly inhibited pAXL expression in various cell lines. SKI‐G‐801 alone or in combination with anti‐PD‐1 potently inhibited metastasis in B16F10 melanoma, CT26 colon and 4T1 breast models. SKI‐G‐801 inhibited the growth of B16F10 and 4T1 tumor‐bearing mice but not immune‐deficient mice. An antibody depletion assay revealed that CD8+ T cells significantly contributed to SKI‐G‐801‐mediated survival. Anti‐PD‐1 and combination group were observed the increased CD8+Ki67+ and effector T cells and M1 macrophage and decreased M2 macrophage, and granulocytic myeloid‐derived suppressor cell (G‐MDSC) compared to the control group. The neoadjuvant combination of SKI‐G‐801 and anti‐PD‐1 therapy achieved superior survival benefits by inducing more profound T‐cell responses in the 4T1 syngeneic mouse model. Conclusion SKI‐G‐801 significantly suppressed tumor metastasis and growth by enhancing anti‐tumor immune responses. Our results suggest that SKI‐G‐801 has the potential to overcome anti‐PD‐1 therapy resistance and allow more patients to benefit from anti‐PD‐1 therapy.

Increased resting metabolic rate (RMR), representing augmented energy expenditure, is a preferred physical characteristic; however, the Tae-Eum Sasang type, with a high incidence of obesity and metabolic diseases, has a higher RMR. This study scrutinized the physical characteristics of Sasang typology, a traditional Korean personalized medicine, to resolve this discrepancy, which can unravel the mechanism of Tae-Eum-type-specific obesity and improve the Tae-Eum Sasang-type diagnosis. A total of 395 healthy participants provided Sasang-type diagnosis using Sasang Constitutional Analysis Tool and physical features, including skeletal muscle mass, body fat mass, and RMR, along with those standardized using body weight. The Tae-Eum-type group showed significantly higher body weight, body mass index, body fat mass, and unstandardized RMR (kcal/day) than others, while their standardized measures of RMR per weigh (RMRw, kcal/day/kg) and percent skeletal muscle (PSM, %) were significantly lower. The logistic regression model revealed that the RMRw is pivotal for discriminating Tae-Eum type from others and explaining the developmental mechanism of Tae-Eum-type obesity. The aforementioned might provide a theoretical framework for Sasang-type diagnosis and Sasang-type-specific health promotion using bodily exercise and medical herbs.

Alzheimer’s disease (AD) is a serious neurodegenerative brain disease that interferes with daily life. The accumulation of beta-amyloid (Aβ), along with oxidative stress-inducing neurocellular apoptosis, has been considered one of the causes of AD. Thus, the purpose of this study is to find natural products that can reduce Aβ accumulation. The ethanol extract of Metasequoia glyptostroboides Hu & Cheng fruits (Cupressaceae) significantly reduced the aggregation of Aβ into oligomers and fibrils determined by Thioflavin T (ThT) assay. The solvent-partitioned ethyl acetate layer was further separated based on the bioassay-guided isolation method combined with the ThT assay. As a result, five compounds were isolated and elucidated as taxoquinone (1), sugiol (2), suginal (3), sandaracopimarinol (4), and sandaracopimaradien-19-ol (5) by comparing NMR data with references. All the compounds significantly reduced the aggregation of Aβ and enhanced the disaggregation of pre-formed Aβ aggregates in a dose-dependent manner. Furthermore, the inhibition of Aβ aggregation by the compounds protected PC12 cells from Aβ aggregate-induced toxicity. Among the five compounds, sandaracopimarinol (4) and sandaracopimaradien-19-ol (5) were the most effective. These results suggest that M. glyptostroboides and isolated five compounds have a potential for further study to be developed as anti-AD agents.

We investigated the effects of resveratrol supplementation on in vitro ruminal fermentation and growth performance of Hanwoo calves. Treatment with three resveratrol concentrations (0%, 0.1%, 0.3%, and 0.5%) was used for in vitro ruminal fermentation. Resveratrol concentrations and pH of rumen fluid were negatively correlated (p < 0.05); therefore, total gas production, total volatile fatty acids, and acetate, propionate, and butyrate levels were significantly higher in the treatments than in the control at all time periods (p < 0.05). The appropriate resveratrol concentration that could be added without negative effects on the rumen was 0.3%. In farm experiments, we divided 14 Korean cattle calves into control (C) and 0.3% resveratrol (T) groups. There were no significant differences in the daily weight gain, feed conversion, final body weight, body length, withers height, and height at hip cross in the resveratrol-fed Hanwoo calves. Immunoglobulin G level was significantly higher in the treatment than in the control (p < 0.05), but IgA and IgM levels did not differ. Supplemental feeding of resveratrol is beneficial to in vitro ruminal fermentation, but it is important to supplement it at 0.3%. Furthermore, resveratrol affects calf immunoglobulin G.

Ruminants are a major source of greenhouse gas emissions, and information on ruminant fermentation and microorganisms is essential to understand ruminant digestion, which is associated with environmental pollution. The present study investigated rumen fermentation and microbial diversity according to the three different growth stages of four Hanwoo steers: growing (12 months, G), early fattening (18 months, EF), and late fattening (25 months, LF). No significant differences were observed in rumen pH and ammonia nitrogen among growth stages. Total volatile fatty acids were significantly higher and propionate and valerate significantly lower in G than in EF and LF (p < 0.05). Ten bacterial phyla were detected, including Firmicutes (47.5–53.5%) and Bacteroidetes (28.4–31.7%), which accounted for 79.2–82.3% of the total bacteria. Prevotella accounted for the highest proportion (31.6–42.6%) of all bacteria in this study but did not differ significantly among the different growth stages. Metaprevotella abundance was significantly higher in G than in the other treatments (p < 0.05). In addition, Paraprevotella tended to be higher in LF than in the other treatments (p = 0.056). Given the differences in the genera of microorganisms with relatively low abundance, additional experiments are needed to determine the effect on fermentation.

During 2016–2018, we collected 3,193 ticks from rural areas in South Korea to investigate the prevalence of severe fever with thrombocytopenia syndrome virus (SFTSV). We detected SFTSV in ticks at an infection rate (IR) of 11.1%. We noted increases in the human IR associated with the monthly SFTSV IR in ticks.

This study was conducted to analyze the diversity census of fecal microbiome in horses using meta-analysis of equine 16S rRNA gene sequences that are available in the Ribosomal Database Project (RDP; Release 11, Update 5). The search terms used were “horse feces (or faeces)” and “equine feces (or faeces)”. A total of 842 sequences of equine feces origin were retrieved from the RDP database, where 744 sequences were assigned to 10 phyla placed within Domain Bacteria. Firmicutes (n = 391) and Bacteroidetes (n = 203) were the first and the second dominant phyla, respectively, followed by Verrucomicrobia (n = 58), Proteobacteria (n = 30) and Fibrobacteres (n = 24). Clostridia (n = 319) was the first dominant class placed within Bacteroidetes while Bacteroidia (n = 174) was the second dominant class placed within Bacteroidetes. The remaining 98 sequences were assigned to phylum Euryarchaeota placed within Domain Archaea, where 74 sequences were assigned to class Methanomicrobia. The current results will improve understanding of the diversity of fecal microbiome in horses and may be used to further analyze equine fecal microbiome in future studies.

Abstract Bees are important pollinators in agriculture. The bee population has recently begun to decline possibly due to pesticides. The bee gut microbiota strongly influences the health of bees. The gut microbiota of bees is composed of distinct members belonging to selective taxa. Chemicals like pesticides can alter the gut microbiota. The present study investigated the effect of carbaryl pesticides on gut microbiota of honey bees, which had come in contact with rapeseed plants (Brassica napus) sprayed with carbaryl wettable powder during the honey bee brood test under semi-field condition. Molecular techniques (conventional and quantitative polymerase chain reaction (PCR), clone library method, and DNA sequencing) were employed to analyze changes in the microbial communities between the pesticide-exposed and unexposed bees. Phylogenetic analysis of 16S rRNA genes of the clones from both groups, showed differences in their respective compositions of core and non-core bacteria. Both groups contained carbohydrate-degrading bacteria such as Gilliamella apicola and Lactobacillus. However, the unexposed bees harbored Alphaproteobacteria, which were absent in the exposed bees. Microorganisms found in honey bee guts such as Snodgrassella alvi and L. kullabergensis, however, were observed only in the exposed bees, but not in the unexposed bees. The difference between the two groups was distinctly recognized when copy numbers of 16S rRNA genes were compared by quantitative PCR. Results showed that the average gene copy number for the unexposed bees was higher than that for the exposed bees. This may indicate the toxic effect of pesticides on bees and gut microbiota.

Background. The acute care surgery (ACS) system is a new model for the prompt management of diseases that require rapid treatment in patients with acute abdomen. This study compared the outcomes and characteristics of the ACS system and traditional on-call system (TROS) for acute appendicitis in South Korea. Methods. This single-center, retrospective study included all patients (aged ≥18 years) who underwent surgery for acute appendicitis in 2016 and 2018. The TROS and ACS system were used for the 2016 and 2018 groups, respectively. We retrospectively obtained data on each patient from the electrical medical records. The independent samples t-test and Mann–Whitney U-test were used for continuous and nonnormally distributed data, respectively. Results. In total, 126 patients were included. The time taken to get from the emergency room admission to the operating room, operation times, and postoperative complication rates were similar between both groups. However, the length of the hospital stay was shorter in the ACS group than in the TROS group (4.3 ± 3.2 days vs. 7.2 ± 9.6 days, p=0.039). Conclusions. Since the introduction of the ACS system, the length of hospital stay for surgical patients has decreased. This may be due to the application of an integrated medical procedure, such as a new clinical pathway, rather than differences in the surgical techniques.

Enteric methane (CH4) emissions produced by microbial fermentation in the rumen resulting in the emission of greenhouse gases (GHG) into the atmosphere. The GHG emissions reduction from the livestock industry can be attained by increasing production efficiency and improving feed efficiency, by lowering the emission intensity of production, or by combining the two. In this work, information was compiled from peer-reviewed studies to analyze CH4 emissions calculated per unit of milk production, energy-corrected milk (ECM), average daily gain (ADG), dry matter intake (DMI), and gross energy intake (GEI), and related emissions to rumen fermentation profiles (volatile fatty acids [VFA], hydrogen [H2]) and microflora activities in the rumen of beef and dairy cattle. For dairy cattle, there was a positive correlation (p < 0.001) between CH4 emissions and DMI (R2 = 0.44), milk production (R2 = 0.37; p < 0.001), ECM (R2 = 0.46), GEI (R2 = 0.50), and acetate/propionate (A/P) ratio (R2 = 0.45). For beef cattle, CH4 emissions were positively correlated (p < 0.05–0.001) with DMI (R2 = 0.37) and GEI (R2 = 0.74). Additionally, the ADG (R2 = 0.19; p < 0.01) and A/P ratio (R2 = 0.15; p < 0.05) were significantly associated with CH4 emission in beef steers. This information may lead to cost-effective methods to reduce enteric CH4 production from cattle. We conclude that enteric CH4 emissions per unit of ECM, GEI, and ADG, as well as rumen fermentation profiles, show great potential for estimating enteric CH4 emissions.

Two isoforms of the 70-kDa ribosomal protein S6 kinase, S6K1 and S6K2, have been identified and are considered key downstream effectors of the mTOR signaling pathway, which is involved in tumor growth and progression. However, their biological roles in the tumor microenvironment are poorly understood. In this study, utilizing tumor xenograft models in S6k1−/− and S6k2−/− mice, we show that loss of S6K1 but not S6K2 in the tumor stroma suppresses tumor growth, accompanied by attenuated tumor angiogenesis. We found that while S6K1 depletion had no effect on the proangiogenic phenotype of endothelial cells, the growth and angiogenesis of tumor xenografts were significantly reduced in wild-type mice upon reconstitution with S6K1-deficient bone marrow cells. Furthermore, upon S6K1 loss, induction of both mRNA and protein levels of Hif-1α and those of the downstream target, Vegf, was compromised in bone marrow–derived macrophages stimulated with lactate. These findings indicate that S6K1 but not S6K2 contributes to establishing a microenvironment that favors tumor growth through mediating angiogenesis, and suggest that attenuated tumor angiogenesis upon loss of S6K1 in the tumor stroma is, at least in part, attributable to impaired upregulation of Vegf in tumor-associated macrophages.

Human epidermal growth factor receptors (HERs) are the primary targets of many directed cancer therapies. However, the reason a specific dimer of HERs generates a stronger proliferative signal than other permutations remains unclear. Here, we used single-molecule immunoprecipitation to develop a biochemical assay for endogenously-formed, entire HER2-HER3 heterodimers. We observed unexpected, large conformational fluctuations in juxta-membrane and kinase domains of the HER2-HER3 heterodimer. Nevertheless, the individual HER2-HER3 heterodimers catalyze tyrosine phosphorylation at an unusually high rate, while simultaneously interacting with multiple copies of downstream signaling effectors. Our results suggest that the high catalytic rate and multi-tasking capability make a concerted contribution to the strong signaling potency of the HER2-HER3 heterodimers.

Abstract Optogenetic stimulation of the peripheral nervous system is a novel approach to motor control, somatosensory transduction, and pain processing. Various optical stimulation tools have been developed for optogenetic stimulation using optical fibers and light-emitting diodes positioned on the peripheral nerve. However, these tools require additional sensors to monitor the limb or muscle status. We present herein a novel optical nerve cuff electrode that uses a single cuff electrode to conduct to simultaneously monitor neural activity and optogenetic stimulation of the peripheral nerve. The proposed optical nerve cuff electrode is designed with a polydimethylsiloxane substrate, on which electrodes can be positioned to record neural activity. We confirm that the illumination intensity and the electrical properties of the optical nerve cuff electrode are suitable for optical stimulation with simultaneous neural activity monitoring in Thy1::ChR2 transgenic mice. With the proposed electrode, the limb status is monitored with continuous streaming signals during the optical stimulation of anesthetized and moving animals. In conclusion, this optical nerve cuff electrode provides a new optical modulation tool for peripheral nervous system studies.

Background: Lung cancer is the leading cause of cancer-related death worldwide. In this study, we used a bioactivity-guided isolation technique to identify constituents of Korean Red Ginseng (KRG) with antiproliferative activity against human lung adenocarcinoma cells. Methods: Bioactivity-guided fractionation and preparative/semipreparative HPLC purification were used with LC/MS analysis to separate the bioactive constituents. Cell viability and apoptosis in human lung cancer cell lines (A549, H1264, H1299, and Calu-6) after treatment with KRG extract fractions and constituents thereof were assessed using the water-soluble tetrazolium salt (WST-1) assay and terminal deoxyribonucleotidyl transferase–mediated dUTP nick end labeling (TUNEL) staining, respectively. Caspase activation was assessed by detecting its surrogate marker, cleaved poly adenosine diphosphate (ADP-ribose) polymerase, using an immunoblot assay. The expression and subcellular localization of apoptosis-inducing factor were assessed using immunoblotting and immunofluorescence, respectively. Results and conclusion: Bioactivity-guided fractionation of the KRG extract revealed that its ethyl acetate–soluble fraction exerts significant cytotoxic activity against all human lung cancer cell lines tested by inducing apoptosis. Chemical investigation of the ethyl acetatesoluble fraction led to the isolation of six ginsenosides, including ginsenoside Rb1 (1), ginsenoside Rb2 (2), ginsenoside Rc (3), ginsenoside Rd (4), ginsenoside Rg1 (5), and ginsenoside Rg3 (6). Among the isolated ginsenosides, ginsenoside Rg3 exhibited the most cytotoxic activity against all human lung cancer cell lines examined, with IC50 values ranging from 161.1 μM to 264.6 μM. The cytotoxicity of ginsenoside Rg3 was found to be mediated by induction of apoptosis in a caspase-independent manner. These findings provide experimental evidence for a novel biological activity of ginsenoside Rg3 against human lung cancer cells. Keywords: Apoptosis, Cytotoxicity, Ginsenoside Rg3, Korean Red Ginseng, Lung cancer

ObjectiveTo evaluate the effect of tamoxifen on female reproductive organs in women with breast cancer.MethodsWe retrospectively reviewed the medical records of 309 women with breast cancer who were currently receiving tamoxifen and undergoing regular gynecological examination.ResultsWe evaluated 92 pre- and 217 postmenopausal women. The prevalence of endometrial thickening was 12% in the pre- and 10.6% in the postmenopausal group. An endometrial biopsy was performed in 43 women and confirmed endometrial cancer in 1, endometrial polyps in 14, and endometrial hyperplasia in 4 women. Transvaginal ultrasonography showed 25 cases of newly developed ovarian cysts. Most ovarian cysts had disappeared during follow-up.ConclusionTamoxifen use in women with breast cancer causes few complications and is considered safe for female reproductive organs in case of regular gynecological examination.

Morchella esculenta (Morchellaceae), one of the most widely appreciated wild edible mushrooms, has demonstrated antitumor activity both in vivo and in vitro. However, the molecular basis underlying its antitumor activity has not yet been elucidated. In this study, we examined the biological effects of the MeOH extract of M. esculenta fruiting bodies on four human lung adenocarcinoma cell lines. Bioactivity-guided fractionation of the MeOH extract followed by chemical investigation of its most cytotoxic hexane-soluble fraction led to the isolation of eight compounds (1 −8), including three fatty acids and five sterols. This is the first report of the isolation of compounds 1 and 3–7 from M. esculenta. Among the eight isolated compounds tested, compounds 1, 3, and 5 exhibited the most potent cytotoxicity to human lung cancer cell lines, with IC50 values ranging from 156.9 to 278 M; this activity was mediated by induction of apoptosis.

Limited information is available regarding the use of microwave-dried Hermetia illucens larvae meal (HILM) as a dietary protein source for broiler diets. Therefore, we investigated the effects of microwave-dried HILM on carcass traits, meat quality, fatty acid (FA) profiles of abdominal fat and meat, and heavy metal residues of the meat in broilers. A total of 126 male broilers were randomly assigned to three dietary treatment groups (6 replicates and 7 birds/pen): a control diet and two experimental diets in which soybean meal was replaced with 25 or 50% HILM. The broilers were slaughtered at 35 days; the carcasses were weighed, and breast and leg meats were excised from 12 birds per treatment (2 birds/pen) for meat analysis. The breast meat quality and proximate composition showed satisfactory results. For the higher HILM diet, the content of saturated FA in the abdominal fat was increased and that of polyunsaturated FA was decreased (p < 0.001); the FA profile of leg meat did not significantly differ between groups. The concentrations of undesirable heavy metals in the HILM and leg meat were below permissible levels. However, the carcass weight was decreased (p < 0.001) in the 50% HILM group. Microwave-dried HILM is a potential ingredient for broiler diets, with up to 25% substitution showing no detrimental effects on carcass traits, meat quality, FA profiles, and heavy metal residues in the meat.

The presence of brown adipocytes within white adipose tissue is associated with phenotypes that exhibit improved metabolism and proper body weight maintenance. Therefore, a variety of dietary agents that facilitate the browning of white adipocytes have been investigated. In this study, we screened a natural product library comprising 133 compounds with the potential to promote the browning of white adipocytes, and found that D-mannitol induces the browning of 3T3-L1 adipocytes by enhancing the expression of brown fat-specific genes and proteins, and upregulating lipid metabolism markers. D-mannitol also increased the phosphorylation of AMP-activated protein kinase (AMPK) and acetyl-CoA carboxylase 1 (ACC), suggesting a possible role in lipolysis and fat oxidation. Moreover, an increase in the expression of genes associated with D-mannitol-induced browning was strongly correlated with the activation of the β3-adrenergic receptor as well as AMPK, protein kinase A (PKA), and PPARγ coactivator 1α (PGC1α). D-mannitol effectively reduced the body weight of mice fed a high-fat diet, and increased the expression of β1-oxidation and energy expenditure markers, such as Cidea, carnitine palmityl transferase 1 (CPT1), uncoupling protein 1 (UCP1), PGC1α, and acyl-coenzyme A oxidase (ACOX1) in the inguinal white adipose tissue. Our findings suggest that D-mannitol plays a dual regulatory role by inducing the generation of a brown fat-like phenotype and enhancing lipid metabolism. These results indicate that D-mannitol can function as an anti-obesity supplement.

Non-alcoholic fatty liver disease (NAFLD) is a leading form of chronic liver disease, with few biomarkers and treatment options currently available. Non-alcoholic steatohepatitis (NASH), a progressive disease of NAFLD, may lead to fibrosis, cirrhosis, and hepatocellular carcinoma. Epigenetic modification can contribute to the progression of NAFLD causing non-alcoholic steatohepatitis (NASH), in which the exact role of epigenetics remains poorly understood. To identify potential therapeutics for NASH, we tested small-molecule inhibitors of the epigenetic target histone methyltransferase EZH2, Tazemetostat (EPZ-6438), and UNC1999 in STAM NASH mice. The results demonstrate that treatment with EZH2 inhibitors decreased serum TNF-alpha in NASH. In this study, we investigated that inhibition of EZH2 reduced mRNA expression of inflammatory cytokines and fibrosis markers in NASH mice. In conclusion, these results suggest that EZH2 may present a promising therapeutic target in the treatment of NASH.

Drug resistance in epithelial ovarian cancer (EOC) is reportedly attributed to the existence of cancer stem cells (CSC), because in most cancers, CSCs still remain after chemotherapy. To overcome this limitation, novel therapeutic strategies are required to prevent cancer recurrence and chemotherapy-resistant cancers by targeting cancer stem cells (CSCs). We screened an FDA-approved compound library and found four voltage-gated calcium channel blockers (manidipine, lacidipine, benidipine, and lomerizine) that target ovarian CSCs. Four calcium channel blockers (CCBs) decreased sphere formation, viability, and proliferation, and induced apoptosis in ovarian CSCs. CCBs destroyed stemness and inhibited the AKT and ERK signaling pathway in ovarian CSCs. Among calcium channel subunit genes, three L- and T-type calcium channel genes were overexpressed in ovarian CSCs, and downregulation of calcium channel genes reduced the stem-cell-like properties of ovarian CSCs. Expressions of these three genes are negatively correlated with the survival rate of patient groups. In combination therapy with cisplatin, synergistic effect was shown in inhibiting the viability and proliferation of ovarian CSCs. Moreover, combinatorial usage of manidipine and paclitaxel showed enhanced effect in ovarian CSCs xenograft mouse models. Our results suggested that four CCBs may be potential therapeutic drugs for preventing ovarian cancer recurrence.

The discovery of small-molecule regulators of microRNAs remains challenging, but a few have been reported. Herein, we describe small-molecule inhibitors of miR-31, a tumor-associated microRNA (miRNA), identified by high-throughput screening using a cell-based reporter assay. Aminosulfonylarylisoxazole compounds exhibited higher specificity for miR-31 than for six other miRNAs, i.e., miR-15a, miR-16, miR-21, miR-92a-1, miR-146a, and miR-155, and increased the expression of miR-31 target genes. The down-regulation of mature miR-31 was observed, while its precursor form increased following treatment with the compounds. Thus, the compounds may target the processing of pre-miR-31 into mature miR-31 and thereby inhibit the production of mature miR-31.

A 51-year-old male patient was referred for a sudden out-of-hospital cardiac arrest. Upon arrival, he was conscious and had no chest pain complaints. There was no abnormality in initial electrocardiographic and echocardiographic examinations. However, episodes of recurrent ventricular fibrillation (VF) were documented on rhythm monitoring. Each VF episode was triggered by an isolated monomorphic ventricular premature complex (VPC). Suspecting idiopathic VF, emergency radiofrequency catheter ablation was planned for the VPCs. However, when coronary angiography was performed to exclude silent ischemia, the results showed a total occlusion of the right coronary artery posterolateral branch, which is thought to supply the left ventricular inferior and septal wall. After successful reperfusion, VF episodes and the triggering VPCs disappeared. We are documenting this case to emphasize the potential for silent myocardial infarction to cause out-of-hospital sudden cardiac arrest even in a patient without any symptom or sign of acute coronary syndrome.

The Me2Si-bridged ansa-Cp/amido half-metallocene, [Me2Si(η5-Me4C5)(NtBu)]TiCl2, termed a “constrained-geometry catalyst (CGC)”, is a representative homogeneous Ziegler catalyst. CGC derivatives with the [1,2]azasilinane framework, in which the amide alkyl substituent is joined by the Si-bridge, were prepared, and the catalytic performances of these species was studied. Me4C5HSi(Me)(CH2CH=CH2)-NH(C(R)(R’)CH=CH2) (R, R’ = H or methyl; Me4C5H = tetramethylcyclopentadienyl) was susceptible to ring closure metathesis (RCM) when treated with Schrock’s Mo-catalyst to afford -Si(Me4C5H)(Me)CH2CH=CHC(R)(R’)NH- containing a six-membered ring framework. Using the precursors and the products of RCM, various CGC derivatives, i.e., [-Si(η5-Me4C5)(Me)CH2CH=CHC(R)(H)N-]TiMe2 (13, R = H; 15, R = Me), [-Si(η5-Me4C5)(Me)CH2CH2CH2CH2N]TiMe2 (14), [(η5-Me4C5)Si(Me)(CH2CH=CH2)NCH2CH=CH2]TiMe2 (16), [(η5-Me4C5)Si (Me)(CH=CH2)NCH2CH=CH2]TiMe2 (17), and [(η5-Me4C5)Si(Me)(CH2CH3)NCH2CH2CH3]TiMe2 (18), were prepared. The catalytic activity of the newly prepared complexes was lower than that of CGC when activated with [Ph3C][B(C6F5)4]/iBu3Al. However, the catalytic activity of these species was improved by using tetrabutylaluminoxane ([iBu2Al]2O) instead of iBu3Al and the activity of 14/[Ph3C][B(C6F5)4]/[iBu2Al]2O was comparable to that of CGC/[Ph3C][B(C6F5)4]/iBu3Al (4.7 and 5.0 × 106 g/mol-Ti, respectively). Advantageously, the newly prepared complexes produced higher molecular weight poly(ethylene-co-1-octene)s than CGC.

Secondary metabolite (SM) production in filamentous fungi is mechanistically associated with chromatin remodeling of specific SM clusters. One locus recently shown to be involved in SM suppression in Aspergillus nidulans was CclA, a member of the histone 3 lysine 4 methylating COMPASS complex. Here we examine loss of CclA and a putative H3K4 demethylase, HdmA, in the human pathogen Aspergillus fumigatus. Although deletion of hdmA showed no phenotype under the conditions tested, the cclA deletant was deficient in tri- and di-methylation of H3K4 and yielded a slowly growing strain that was rich in the production of several SMs, including gliotoxin. Similar to deletion of other chromatin modifying enzymes, ΔcclA was sensitive to 6-azauracil indicating a defect in transcriptional elongation. Despite the poor growth, the ΔcclA mutant had wild-type pathogenicity in a murine model and the Toll-deficient Drosophila model of invasive aspergillosis. These data indicate that tri- and di-methylation of H3K4 is involved in the regulation of several secondary metabolites in A. fumigatus, however does not contribute to pathogenicity under the conditions tested.

Purpose: By utilizing both protein and mRNA expression patterns, we can identify more detailed and diverse immune cells, providing insights into understanding the complex immune landscape in cancer ecosystems. Materials and Methods: This study was performed by obtaining publicly available Cellular Indexing of Transcriptomes and Epitopes by Sequencing (CITE-seq) data of peripheral blood mononuclear cells (PBMCs) from the Gene Expression Omnibus database. A total of 94674 total cells were analyzed, of which 32412 were T cells. There were 228 protein features and 16262 mRNA features in the data. The Seurat package was used for quality control and preprocessing, principal component analysis was performed, and Uniform Manifold Approximation and Projection was used to visualize the clusters. Protein and mRNA levels in the CITE-seq were analyzed. Results: We observed that a subset of T cells in the clusters generated at the protein level divided better. By identifying mRNA markers that were highly correlated with the CD4 and CD8 proteins and cross-validating CD26 and CD99 markers using flow cytometry, we found that CD4+ and CD8+ T cells were better discriminated in PBMCs. Weighted Nearest Neighbor clustering results identified a previously unobserved T cell subset. Conclusion: In this study, we used CITE-seq data to confirm that protein expression patterns could be used to identify cells more precisely. These findings will improve our understanding of the heterogeneity of immune cells in the future and provide valuable insights into the complexity of the immune response in health and disease. [ABSTRACT FROM AUTHOR]