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The rapid advancements in nucleic acid-based electrochemical sensors for implantable and wearable applications have marked a significant leap forward in the domain of personal healthcare over the last decade. This technology promises to revolutionize personalized healthcare by facilitating the early diagnosis of diseases, monitoring of disease progression, and tailoring of individual treatment plans. This review navigates through the latest developments in this field, focusing on the strategies for nucleic acid sensing that enable real-time and continuous biomarker analysis directly in various biofluids, such as blood, interstitial fluid, sweat, and saliva. The review delves into various nucleic acid sensing strategies, emphasizing the innovative designs of biorecognition elements and signal transduction mechanisms that enable implantable and wearable applications. Special perspective is given to enhance nucleic acid-based sensor selectivity and sensitivity, which are crucial for the accurate detection of low-level biomarkers. The integration of such sensors into implantable and wearable platforms, including microneedle arrays and flexible electronic systems, actualizes their use in on-body devices for health monitoring. We also tackle the technical challenges encountered in the development of these sensors, such as ensuring long-term stability, managing the complexity of biofluid dynamics, and fulfilling the need for real-time, continuous, and reagentless detection. In conclusion, the review highlights the importance of these sensors in the future of medical engineering, offering insights into design considerations and future research directions to overcome existing limitations and fully realize the potential of nucleic acid-based electrochemical sensors for healthcare applications.
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Técnicas Biossensoriais , Técnicas Eletroquímicas , Ácidos Nucleicos , Dispositivos Eletrônicos Vestíveis , Humanos , Ácidos Nucleicos/análise , Biomarcadores/análise , Biomarcadores/sangueRESUMO
The multifunctional carbon catabolite repression negative on TATA-box-less complex (CCR4-NOT) is a multi-subunit complex present in all eukaryotes, including fungi. This complex plays an essential role in gene expression; however, a functional study of the CCR4-NOT complex in the rice blast fungus Magnaporthe oryzae has not been conducted. Seven genes encoding the putative CCR4-NOT complex were identified in the M. oryzae genome. Among these, a homologous gene, MoNOT3, was overexpressed during appressorium development in a previous study. Deletion of MoNOT3 in M. oryzae resulted in a significant reduction in hyphal growth, conidiation, abnormal septation in conidia, conidial germination, and appressorium formation compared to the wild-type. Transcriptional analyses suggest that the MoNOT3 gene affects conidiation and conidial morphology by regulating COS1 and COM1 in M. oryzae. Furthermore, Δmonot3 exhibited a lack of pathogenicity, both with and without wounding, which is attributable to deficiencies in the development of invasive growth in planta. This result was also observed in onion epidermal cells, which are non-host plants. In addition, the MoNOT3 gene was involved in cell wall stress responses and heat shock. Taken together, these observations suggest that the MoNOT3 gene is required for fungal infection-related cell development and stress responses in M. oryzae.
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Ascomicetos , Magnaporthe , Oryza , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Ascomicetos/metabolismo , Esporos Fúngicos , Oryza/genética , Doenças das Plantas/genética , Doenças das Plantas/microbiologia , Regulação Fúngica da Expressão GênicaRESUMO
Calaphidinae is the second-largest subfamily in the family Aphididae. Despite their species diversity and some taxonomic controversy, no phylogenetic studies have been conducted on them thus far. Herein, we report the first molecular phylogeny of Calaphidinae and two related lineages, Phyllaphidinae and Saltusaphidinae, based on five genes (3418 bp) for 126 taxa. Maximum parsimony, maximum-likelihood and Bayesian inference phylogenetic analyses were performed on the multilocus dataset. Divergence time estimation, biogeographical reconstruction, ancestral host plant reconstruction and PhyloType analyses were performed to identify evolutionary trends in Calaphidinae. Our phylogenetic results lead to several conclusions: Phyllaphidinae is a sister group to Calaphidinae s.l.; Calaphidinae is paraphyletic with respect to the former "Saltusaphidinae"; the ingroup clade was subdivided into nine newly recognized lineages; and three subtribes of Calaphidinae (Monaphidina, Calaphdina and Panaphidina) and many genera were not recovered as monophyletic. A new classification is proposed with eight tribal divisions that reflect our phylogenetic results, including three new tribes (Pterocallidini trib.n., Pseudochromaphidini trib.n. and Shivaphidini trib.n.) and three new statuses (Saltusaphidini stat.n., Therioaphidini stat.n. and Myzocallidini stat.n.). The ancestral reconstruction results imply that the ingroup taxa's common ancestor originated in the Eastern Palaearctic and might have fed on Fagaceae in the Late Cretaceous. Later, multiple host shifts and an expanding geographical distribution led to the current species diversity of Calaphidinae. Our reconstructions suggest that species diversification cannot solely be explained by speciation via host shifts and that geographical isolation probably also played a key role. Our results provide new insight into the natural classification and history of the host plant associations and biogeography of Calaphidinae s.l.
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Afídeos , Animais , Afídeos/genética , Teorema de Bayes , Geografia , FilogeniaRESUMO
ABSTRACT: From Operation Warp Speed to the lipid mRNA vaccine, the COVID-19 pandemic has been a watershed moment for technological development, production, and implementation. The scale and pace of innovation and global collaboration has likely not been experienced since World War II. This article highlights some of the engineering accomplishments that occurred during the pandemic. We provide a broad overview of the technological achievements in vaccine design, antibody engineering, drug repurposing, and rapid diagnostic testing. We also discuss what the future of these technologies and the future of large-scale collaborations might look like moving forward.
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Pears (Pyrus pylifolia L.) are cultivated nationwide as one of the most economically important fruit trees in Korea. At the end of October 2019, bleeding canker was observed in a pear orchard located in Naju, Jeonnam Province (34°53'50.54â³ N, 126°39'00.32â³ E). The canker was observed on trunks and branches of two 25-year-old trees, and the diseased trunks and branches displayed partial die-back or complete death. When the bark was peeled off from the diseased trunks or branches, brown spots or red streaks were found in the trees. Bacterial ooze showed a rusty color and the lesion was sap-filled with a yeasty smell. Trunks displaying bleeding symptoms were collected from two trees. Infected bark tissues (3 × 3 mm) from the samples were immersed in 70% ethanol for 1 minute, rinsed three times in sterilized water, ground to fine powder using a mortar and pestle, and suspended in sterilized water. After streaking each suspension on Luria-Bertani (LB) agar, the plates were incubated at 25°C without light for 2 days. Small yellow-white bacterial colonies with irregular margins were predominantly obtained from all the samples. Three representative isolates (ECM-1, ECM-2 and ECM-3) were subjected to further characterization. These isolates were cultivated at 39 ï°C, and utilized (-)-D-arabinose, (+) melibiose, (+)raffinose, mannitol and myo-inositol but not 5-keto-D-gluconate, ï¢-gentiobiose, or casein. These isolates were identified as Dickeya sp. based on the sequence of 16S rRNA (MT820458-820460) gene amplified using primers 27f and 1492r (Heuer et al. 2000). The 16S rRNA sequences matched with D. fangzhongdai strain ND14b (99.93%; CP009460.1) and D. fangzhongdai strain PA1(99.86%; CP020872.1). The recA, fusA, gapA, purA, rplB, and dnaX genes and the intergenic spacer (IGS) regions were also sequenced as described in Van der wolf et al. (2014). The recA (MT820437-820439), fusA (MT820440-820442), gapA (MT820443-820445), purA (MT820446-820448), rplB (MT820449-820451), dnaX (MT820452-820454) and IGS (MT820455-820457) sequences matched with D. fangzhongdai strains JS5, LN1 and QZH3 (KT992693-992695, KT992697-992699, KT992701-992703, KT992705-992707, KT992709-992711, KT992713-992715, and KT992717-992719, respectively). A neighbor-joining phylogenetic analysis based on the concatenated recA, fusA, gapA, purA, rplB, dnaX and IGS sequences placed the representative isolates within a clade comprising D. fangzhongdai. ECM-1 to 3 were grouped into a clade with one strain isolated from waterfall, D. fangzhongdai ND14b from Malaysia. Pathogenicity test was performed using isolate ECM-1. Three two-year-old branches and flower buds on 10-year-old pear tree (cv. Nittaka), grown at the National Institute of Horticultural and Herbal Science Pear Research Institute (Naju, Jeonnam Province in Korea), were inoculated with 10 µl and 2 µl of a bacterial suspension (108 cfu/ml), respectively, after wounding inoculation site with a sterile scalpel (for branch) or injecting with syringe (for flower bud). Control plants were inoculated with water. Inoculated branches and buds in a plastic bag were placed in a 30â incubator without light for 2 days (Chen et al. 2020). Both colorless and transparent bacterial ooze and typical bleeding canker were observed on both branches and buds at 3 and 2 weeks post inoculation, respectively. No symptoms were observed on control branches and buds. This pathogenicity assay was conducted three times. We reisolated three colonies from samples displaying the typical symptoms and checked the identity of one by sequencing the dnaX locus. Dickeya fangzhongdai has been reported to cause bleeding canker on pears in China (Tian et al. 2016; Chen et al. 2020). This study will contribute to facilitate identification and control strategies of this disease in Korea. This is the first report of D. fangzhongdai causing bleeding canker on pears in Korea.
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Far-infrared rays (FIR) are known to have various effects on atoms and molecular structures within cells owing to their radiation and vibration frequencies. The present study examined the effects of FIR on gene expression related to glucose transport through microarray analysis in rat skeletal muscle cells, as well as on mitochondrial biogenesis, at high and low glucose conditions. FIR were emitted from a bio-active material coated fabric (BMCF). L6 cells were treated with 30% BMCF for 24 h in medium containing 25 or 5.5 mM glucose, and changes in the expression of glucose transporter genes were determined. The expression of GLUT3 (Slc2a3) increased 2.0-fold (p < 0.05) under 5.5 mM glucose and 30% BMCF. In addition, mitochondrial oxygen consumption and membrane potential (ΔΨm) increased 1.5- and 3.4-fold (p < 0.05 and p < 0.001), respectively, but no significant change in expression of Pgc-1a, a regulator of mitochondrial biogenesis, was observed in 24 h. To analyze the relationship between GLUT3 expression and mitochondrial biogenesis under FIR, GLUT3 was down-modulated by siRNA for 72 h. As a result, the ΔΨm of the GLUT3 siRNA-treated cells increased 3.0-fold (p < 0.001), whereas that of the control group increased 4.6-fold (p < 0.001). Moreover, Pgc-1a expression increased upon 30% BMCF treatment for 72 h; an effect that was more pronounced in the presence of GLUT3. These results suggest that FIR may hold therapeutic potential for improving glucose metabolism and mitochondrial function in metabolic diseases associated with insufficient glucose supply, such as type 2 diabetes.
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The novel isolate belonging to the genus Terrimonas, designated CR94T, was isolated from rhizosphere soil of a ginseng field in Geumsan, Korea. Cells of strain CR94T were strictly aerobic, Gram-stain-negative, non-motile, non-filamentous single rods. Growth was observed at 10-37 °C (optimum 28 °C) and at pH 4.0-10.0 (optimum pH 6.0). Phylogenetic analyses based on 16S rRNA gene sequences indicated that strain CR94T belonged to the genus Terrimonas, showing highest sequence similarity to Terrimonas lutea DYT (97.3â%), Terrimonas pekingensis QHT (97.1â%), Terrimonas aquatica RIB1-6T (95.6â%), Terrimonas rubra M-8T (94.7â%) and Terrimonas ferruginea ATCC 13524T (93.8â%). DNA-DNA relatedness values between strain CR94T and T. lutea KACC 13047T, T. pekingensis KACC 18795T, T. ferruginea KACC 11310T and T. aquatica LMG 24825T were 30.5, 28.9, 17.8 and 13.5â%, respectively. The DNA G+C content was 46.5 mol% and the major respiratory quinone was menaquinone-7 (MK-7). The major cellular fatty acids of strain CR94T were iso-C15:1 G and iso-C15â:â0. On the basis of the polyphasic analysis, strain CR94T represents a novel species of the genus Terrimonas, for which the name Terrimonas rhizosphaerae sp. nov. is proposed. The type strain is CR94T (=KACC 17564T=NCAIM B 025317T).
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Bacteroidetes/classificação , Panax/microbiologia , Filogenia , Rizosfera , Microbiologia do Solo , Técnicas de Tipagem Bacteriana , Bacteroidetes/genética , Bacteroidetes/isolamento & purificação , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/química , Hibridização de Ácido Nucleico , RNA Ribossômico 16S/genética , República da Coreia , Análise de Sequência de DNA , Vitamina K 2/análogos & derivados , Vitamina K 2/químicaRESUMO
A denitrifying bacterium, designated strain E4-1(T), was isolated from a bioreactor for tannery wastewater treatment, and its taxonomic position was investigated using a polyphasic approach. Strain E4-1(T), a facultative anaerobic bacterium, was observed to grow between 0 and 12 % (w/v) NaCl, between pH 3.0 and 12.0. Cells were found to be oxidase-positive and catalase-negative. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain E4-1(T) forms a distinct lineage with respect to closely related genera in the family Xanthomonadaceae, and is closely related to Chiayiivirga, Aquimonas and Dokdonella, and the levels of 16S rRNA gene sequence similarity with respect to the type species of related genera are less than 93.9 %. The predominant respiratory quinone was determined to be ubiquinone-8 (Q-8) and the major cellular fatty acids were determined to be iso-C15:0, iso-C17:1 ω9c, iso-C11:0 and iso-C11:0 3OH. Based on physiological, biochemical and chemotaxonomic properties together with results of comparative 16S rRNA gene sequence analysis, strain E4-1(T) is considered to represent a novel species in a new genus, for which the name Denitratimonas tolerans gen. nov., sp. nov. is proposed. The type strain is E4-1(T) (=KACC 17565(T) = NCAIM B 025327(T)).
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Reatores Biológicos/microbiologia , Águas Residuárias/microbiologia , Xanthomonadaceae/classificação , Xanthomonadaceae/isolamento & purificação , DNA Bacteriano/genética , DNA Ribossômico/genética , Desnitrificação , Ácidos Graxos/metabolismo , Fosfolipídeos/metabolismo , Filogenia , Quinonas/metabolismo , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Cloreto de Sódio/metabolismo , Ubiquinona/metabolismo , Xanthomonadaceae/genética , Xanthomonadaceae/metabolismoRESUMO
The tribe Tuberolachnini within the Lachninae (Hemiptera: Aphididae) is particularly intriguing due to its morphological traits and various ecological associations. Among the genera within this group, Pyrolachnus stands out as relatively understudied. Currently, only one species, Pyrolachnus imbricatus nipponicus Sorin, 2011, is known from Japan, distinguished by its distinctive characteristics. Through meticulous morphological analyses, we introduce a novel Lachninae genus, Miyalachnusgen. nov., associated with Cerasus and Prunus spp. (Rosaceae) in Japan. This new genus accommodates P. imbricatus nipponicus, now recognized as Miyalachnus nipponicus (Sorin, 2011) comb. nov. Additionally, we present a second species within this genus, Miyalachnus sorinisp. nov., along with comprehensive SEM morphological examination and insights into its biology. Our study describes in detail the morphological characteristics of both viviparous and bisexual generations of Miyalachnus, as well as their relationships with related genera.
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Citizen science (CS) is the most effective tool for overcoming the limitations of government and/or professional data collection. To compensate for quantitative limitations of the 'Winter Waterbird Census of Korea', we conducted a total of four bird monitoring via CS from 2021 to 2022. To use CS data alongside national data, we studied CS data quality and improvement utilizing (1) digit-based analysis using Benford's law and (2) comparative analysis with national data. In addition, we performed bird community analysis using CS-specific data, demonstrating the necessity of CS. Neither CS nor the national data adhered to Benford's law. Alpha diversity (number of species and Shannon index) was lower, and total beta diversity was higher for the CS data than national data. Regarding the observed bird community, the number of species per family was similar; however, the number of individuals per family/species differed. We also identified the necessity of CS by confirming the possibility of predicting bird communities using CS-specific data. CS was influenced by various factors, including the perceptions of the survey participants and their level of experience. Therefore, conducting CS after systematic training can facilitate the collection of higher-quality data.
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Aves , Censos , Ciência do Cidadão , Animais , Aves/fisiologia , República da Coreia , BiodiversidadeRESUMO
BACKGROUND AND PURPOSE: To investigate the molecular mechanism by which irradiated macrophages secrete cytosolic double-stranded DNA (c-dsDNA) to increase radiosensitivity of tumors. MATERIALS AND METHODS: Irradiated bone marrow-derived macrophages (BMDM) were co-incubated with irradiated EO771 or MC38 cancer cells to determine clonogenic survival. c-dsDNA were measured by agarose gel or enzyme-linked immunosorbent assay. BMDM or cancer cells were analyzed with immunostaining or western blot. Subcutaneously implanted MC38 cells in myeloid-specific Prkdc knockout (KO) mice or littermate control mice were irradiated with 8 Gy to determine radiosensitivity of tumors. RESULTS: We observed that irradiated BMDM significantly increased radiosensitivity of cancer cells. By performing immunostaining, we found that there was a dose-dependent increase in the formation of c-dsDNA and phosphorylation in DNA-dependent protein kinase (DNA-PK) in irradiated BMDM. Importantly, c-dsDNA in irradiated BMDM could be secreted to the extracellular milieu and this process required DNA-PK, which phosphorylated myosin light chain to regulate the secretion. The secreted c-dsDNA from irradiated BMDM then activated toll-like receptor-9 and subsequent nuclear factor kappa-light-chain-enhancer of activated B cells signaling in the adjacent cancer cells inhibiting radiation-induced DNA double strand break repair. Lastly, we observed that irradiated tumors in vivo had a significantly increased number of tumor-associated macrophages (TAM) with phosphorylated DNA-PK expression in the cytosol. Furthermore, tumors grown in myeloid-specific Prkdc KO mice, in which TAM lacked phosphorylated DNA-PK expression were significantly more radioresistant than those of the wild-type control mice. CONCLUSIONS: Irradiated macrophages can increase antitumor efficacy of radiotherapy through secretion of c-dsDNA under the regulation of DNA-PK.
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Proteína Quinase Ativada por DNA , Neoplasias , Camundongos , Animais , Citosol/metabolismo , Tolerância a Radiação , Macrófagos , DNARESUMO
Fertilizers are widely used to improve the quality of fruits and vegetables. However, the overuse of fertilizers has become an issue because it causes environmental problems and negatively affects productivity and fruit quality. In this study, we examined the effects of nitrogen, phosphorus, and potassium (NPK) fertilizer levels on the metabolism of cucumber fruit in low- and high-nutrient soils using mass-spectrometry-based metabolomics approaches. Cucumber metabolite content was notably different depending on the initial soil nutrient status. Most amino acids and phenylpropanoids were abundant in the cucumbers raised in low-nutrient soil, whereas organic acids, some amino acids (aspartate, glutamate, and ornithine), and carbohydrates were comparatively higher in fruits from high-nutrient soil. The fertilizer supply resulted in an alteration in the metabolite profile, while no change in fruit yield was observed in either low- or high-nutrient soils. Fertilizer treatment perturbed the metabolite contents in cucumbers from low-nutrient soil. In contrast, treatment with higher concentrations of fertilizer in high-nutrient soil increased phenylpropanoid content in the cucumbers, while most metabolites decreased. In conclusion, fertilization levels should be carefully determined, considering culture conditions such as the original soil status, to increase product yield and fruit quality and avoid environmental problems.
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BACKGROUND: Emergency departments (ED) nurses experience high mental workloads because of unpredictable work environments; however, research evaluating ED nursing workload using a tool incorporating nurses' perception is lacking. Quantify ED nursing subjective workload and explore the impact of work experience on perceived workload. METHODS: Thirty-two ED nurses at a tertiary academic hospital in the Republic of Korea were surveyed to assess their subjective workload for ED procedures using the National Aeronautics and Space Administration Task Load Index (NASA-TLX). Nonparametric statistical analysis was performed to describe the data, and linear regression analysis was conducted to estimate the impact of work experience on perceived workload. RESULTS: Cardiopulmonary resuscitation (CPR) had the highest median workload, followed by interruption from a patient and their family members. Although inexperienced nurses perceived the 'special care' procedures (CPR and defibrillation) as more challenging compared with other categories, analysis revealed that nurses with more than 107 months of experience reported a significantly higher workload than those with less than 36 months of experience. CONCLUSION: Addressing interruptions and customizing training can alleviate ED nursing workload. Quantified perceived workload is useful for identifying acceptable thresholds to maintain optimal workload, which ultimately contributes to predicting nursing staffing needs and ED crowding.
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Serviço Hospitalar de Emergência , Carga de Trabalho , Humanos , Carga de Trabalho/psicologia , Serviço Hospitalar de Emergência/organização & administração , Feminino , Masculino , República da Coreia , Adulto , Inquéritos e Questionários , Enfermagem em Emergência , Pessoa de Meia-Idade , Análise e Desempenho de TarefasRESUMO
Monocytes are members of the mononuclear phagocyte system involved in pathogen clearance and nanoparticle pharmacokinetics. Monocytes play a critical role in the development and progression of cardiovascular disease and, recently, in SARS-CoV-2 pathogenesis. While studies have investigated the effect of nanoparticle modulation on monocyte uptake, their capacity for nanoparticle clearance is poorly studied. In this study, we investigated the impact of ACE2 deficiency, frequently observed in individuals with cardiovascular complications, on monocyte nanoparticle endocytosis. Moreover, we investigated nanoparticle uptake as a function of nanoparticle size, physiological shear stress, and monocyte phenotype. Our Design of Experiment (DOE) analysis found that the THP-1 ACE2 - cells showed a greater preference for 100nm particles under atherosclerotic conditions than THP-1 wild-type cells. Observing how nanoparticles can modulate monocytes in the context of disease can inform precision dosing.
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In the rice processing industry, wastewater is an inevitable by-product of rice washing. To increase the utilization of washed rice water (WRW), seven types of fermented washed rice water (FWRW) were prepared using lactic acid bacteria (LAB) and carbohydrate hydrolase. The total concentration of small maltooligosaccharides (MOSs) in the amyloglucosidase (AMG) treatment groups was about ten times higher than in the untreated groups. After 6 h of fermentation, six of the seven FWRW samples reached a pH of 4 due to the increased concentration of organic acids and could, therefore, be used as food acidity regulators. To confirm the applicability of FWRW, the traditional Korean rice cake garaetteok was prepared with FWRW and stored at 4 °C for 5 days. A texture profile analysis (TPA) revealed that the hardness of garaetteok treated with FWRW was significantly lower than that of untreated garaetteok following storage. Differential scanning calorimetry (DSC) showed that FWRW retarded the retrogradation of garaetteok during storage. The addition of FWRW using Lactobacillus reuteri with an AMG group was particularly effective for inhibiting microbial activity in garaetteok during storage. These results suggest that FWRW using AMG-added L. reuteri can be used as a novel food additive for improving the quality of traditional Korean starch foods and could also reduce the volume of waste WRW.
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Amylopectin clusters (APCs) are produced by cyclodextrin glucanotransferase (EC 2.4.1.19). Their solubility rate in aqueous solution was found to be 16.7 %. The weight-average molecular weight of APCs is â¼105 Da, as determined by multiangle laser light scattering analysis. Side chain length analysis indicated that the relative proportions of side chains with a degree of polymerization in the ranges of 2-8 and 25-50 decreased and increased, respectively, during preparation of APCs. In the exercise experiment, the blood glucose level of rats was higher in the APC-treated group than in the groups treated with commercial carbohydrate supplement (CCD) and glucose. In the forced swimming test, the swimming time in the APC and CCD groups increased by 22.6 % and 31.1 %, respectively, compared with the glucose administration group. The insulin levels were also similar between the APC and CCD groups. However, the glycogen levels in the liver and muscles of mice were significantly higher in the APC group than control group. These results suggest that APCs could potentially enhance endurance when added to sports drinks.
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Environmental DNA (eDNA) extracted from the gut contents of filter feeders can be used to identify biodiversity in aquatic ecosystems. In this study, we used eDNA from the gut contents of the Asian clam Corbicula fluminea to examine biodiversity within estuarine ecosystem. Field sampling was conducted at three points in the Nakdong River Estuary, which is characterised by closed estuarine features resulting from the presence of an estuarine barrage. The collected C. fluminea samples were dissected to separate the gut contents, and the extracted eDNA was amplified using 18S V9 primer targeting all eukaryote-derived DNA. The amplified DNA was sequenced using a next-generation sequencing (NGS) technique, and a BLASTn search was performed based on the National Centre for Biotechnology Information (NCBI) database for taxa identification. We obtained 23 unique operational taxonomic units (OTUs), including fish (approximately 8.70%), copepods (approximately 17.39%), and green algae (approximately 21.74%), representing a wide range of habitats. Furthermore, 8 out of the 20 families were identified through comparisons with reference data from conventional field surveys, and the OTUs of elusive migratory fish were detected. The results support the application of C. fluminea as an eDNA sampler for supplementary biodiversity monitoring.
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Tauopathies are neurodegenerative diseases characterized by abnormal conformational changes in tau protein. Early hyperphosphorylation-induced conformational changes are considered a hallmark of tauopathy, but real-time tracking methods are lacking. Here, we present two novel fluorescence resonance energy transfer (FRET)-based tau biosensors that detect such changes with high spatiotemporal resolution at the single-cell level. The TAUCON biosensor measures instantaneous conformational changes in hyperphosphorylated tau within 20 min, while the TAUCOM biosensor detects changes in the paper-clip structure of microtubule-associated tau. Our biosensors provide faster and more precise detection than conventional methods and can serve as valuable tools for investigating the initial causes, mechanisms, progression, and treatment of tauopathies.
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Técnicas Biossensoriais , Doenças Neurodegenerativas , Tauopatias , Humanos , Proteínas tau/metabolismo , Transferência Ressonante de Energia de Fluorescência/métodos , Tauopatias/diagnóstico , Tauopatias/metabolismoRESUMO
DNA-dependent protein kinase catalytic subunit (DNA-PKcs), a member of the phosphatidylinositol 3-kinase related kinase family is a well-known player in repairing DNA double strand break through non-homologous end joining pathway. This mechanism has allowed us to understand its critical role in T and B cell development through V(D)J recombination and class switch recombination, respectively. We have also learned that the defects in these mechanisms lead to severely combined immunodeficiency (SCID). Here we highlight some of the latest evidence where DNA-PKcs has been shown to localize not only in the nucleus but also in the cytoplasm, phosphorylating various proteins involved in cellular metabolism and cytokine production. While it is an exciting time to unveil novel functions of DNA-PKcs, one should carefully choose experimental models to study DNA-PKcs as the experimental evidence has been shown to differ between cells of defective DNA-PKcs and those of DNA-PKcs knockout. Moreover, while there are several DNA-PK inhibitors currently being evaluated in the clinical trials in attempt to increase the efficacy of radiotherapy or chemotherapy, multiple functions and subcellular localization of DNA-PKcs in various types of cells may further complicate the effects at the cellular and organismal level.
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Proteína Quinase Ativada por DNA , Proteínas de Ligação a DNA , Proteína Quinase Ativada por DNA/genética , Proteína Quinase Ativada por DNA/metabolismo , Proteínas de Ligação a DNA/metabolismo , Domínio Catalítico , Reparo do DNA , DNARESUMO
To improve the applicability of quercetin (QCT), we produced a QCT and cycloamylose (CA-QCT) inclusion complex based on the cyclization activity of cyclodextrin glucanotransferase (CGTase; EC 2.4.1.19). The encapsulated QCT was purified using recycling preparative high-performance liquid chromatography, and its formation was analyzed using matrix-assisted laser desorption/ionization-time of flight mass spectrometry. The water solubility of CA-QCT was 55,000-fold higher than that of QCT. CA-QCT had 97 % stability for one week at pH 8 in a 4 °C water bath. According to a 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity assay, CA-QCT activity in aqueous solution was 24 times higher than that of an equal amount of QCT in aqueous solution. In an anti-inflammatory assay using lipopolysaccharide-induced RAW264.7 macrophages, CA-QCT in aqueous solution decreased nitric oxide production in a similar manner to QCT in dimethyl sulfoxide (DMSO). Additionally, even under aqueous conditions, CA-QCT more effectively inhibited the production of inflammatory mediators, such as interleukin-1ß, interleukin-6, and cyclooxygenase, compared with QCT dissolved in DMSO.