ABSTRACT
The method presented herein is associated with the Lab Resource article titled "Generation of αMHC-EGFP knock-in in human pluripotent stem cell line, SNUe003-A-3, using CRISPR/CAS9-based gene targeting" [1]. The cardiac muscle-specific protein, α-myosin heavy chain (αMHC), is encoded by the human MYH6 gene, which is expressed in both the atria and ventricles during embryonic development and is predominantly expressed in the atria after birth [2]. Herein, the methods used to achieve CRISPR/SpCas9-mediated introduction of an EGFP reporter into αMHC, the target locus in human pluripotent stem cells (hPSCs) for cardiac lineage tracing and clinical cell sorting are described. The CRISPR-Cas9 system enables efficient replacement of the stop codon in the last exon of αMHC with a 2A non-joining peptide (T2A)-EGFP cassette. First, hPSCs are transfected with the donor construct and Cas9/sgRNA plasmids via electroporation and selected with neomycin for approximately 3 weeks. Thereafter, the established cell line exhibits typical characteristics of human embryonic stem cells (hESCs). When these cells differentiate into cardiomyocytes, the expression of EGFP is confirmed using confocal microscopy, flow cytometry analysis, and immunostaining.â¢The line enables monitoring of cell maturation events during human cardiac development.â¢The line is a valuable platform for cardiotoxicity tests and drug screening.â¢This method has already been employed in two original studies, as previously reported for reporter cell line generation using CRISPR/Cas9.
ABSTRACT
Rice, one of the most widely consumed staples worldwide, serves as a versatile gluten-free substitute. However, review articles on technological developments in grain-free production focusing on rice are scarce. This review assesses various research results concerning the quality attributes of rice-based gluten-free foods, including bread, pasta, and beer. To optimize the key attributes in processed products, such as dough leavening in bread and the physical and cooking properties of noodles and pasta, research has focused on blending different gluten-free grains and incorporating additives that mimic the gluten function. Additionally, various processing technologies, such as starch preprocessing and extrusion puffing processes, have been employed to boost the quality of rice-based gluten-free products. Today, a variety of products, including bread, noodles, and beer, use rice as a partial replacement for barley or wheat. With rapid advancements in technology, a noticeable portion of consumers now shows a preference for products containing rice as a substitute. This trend indicates that rice-based gluten-free foods can be enhanced by leveraging the latest developments in gluten-free product technologies, particularly in countries where rice is a staple or is predominantly cultivated.
ABSTRACT
Boron nitride nanotubes (BNNTs) were purified without the use of a dispersant by controlling the surface tension and steric repulsion of solvent molecules. This method effectively enhanced the difference in solubilities of impurities and BNNTs. The purification process involved optimizing the alkyl-chains of alcohol solvents and adjusting the concentration of alcohol solvent in water to regulate surface tension and steric repulsion. Among the solvents tested, a 70 wt% t-butylalcohol in water mixture exhibited the highest selective isolation of BNNTs from impurities based on differences in solubilities. This favorable outcome was attributed to the surface tension matching with BNNTs, steric repulsion from bulky alkyl chain structures, and differences in interfacial energy between BNNT-liquid and impurity-liquid interfaces. Through this optimized purification process, impurities were removed to an extent of up to 93.3%. Additionally, the purified BNNTs exhibited a distinct liquid crystal phase, which was not observed in the unpurified BNNTs.
ABSTRACT
Microfiber fabrication via wet-spinning of lyotropic liquid crystals (LCs) with anisotropic nanomaterials has gained increased attention due to the microfibers' excellent physical/chemical properties originating from the unidirectional alignment of anisotropic nanomaterials along the fiber axis with high packing density. For wet-spinning of the microfibers, however, preparing lyotropic LCs by achieving high colloidal stability of anisotropic nanomaterials, even at high concentrations, has been a critically unmet prerequisite, especially for recently emerging nanomaterials. Here, we propose a cationically charged polymeric stabilizer that can efficiently be adsorbed on the surface of boron nitride nanotubes (BNNTs), which provide steric hindrance in combination with Coulombic repulsion leading to high colloidal stability of BNNTs up to 22 wt %. The BNNT LCs prepared from the dispersions with various stabilizers were systematically compared using optical and rheological analysis to optimize the phase behavior and rheological properties for wet-spinning of the BNNT LCs. Systematic optical and mechanical characterizations of the BNNT microfibers with aligned BNNTs along the fiber axis revealed that properties of the microfibers, such as their tensile strength, packing density, and degree of BNNT alignment, were highly dependent on the quality of BNNT LCs directly related to the types of stabilizers.
ABSTRACT
TUBB3 is a structural neuronal protein important for multiple neuronal functions including axonal guidance and maturation. This study aimed to generate a human pluripotent stem cell (hPSC) line with a TUBB3-mCherry reporter using CRISPR/SpCas9 nuclease. The stop codon in the last exon of TUBB3 was replaced with a T2A-mCherry cassette using CRISPR/SpCas9-mediated homologous recombination. The established TUBB3-mCherry knock-in cell line exhibited typical pluripotent characteristics. The mCherry reporter faithfully replicated the endogenous level of TUBB3 upon induction of neuronal differentiation. The reporter cell line could contribute to the investigation of neuronal differentiation, neuronal toxicity, and neuronal tracing.
Subject(s)
CRISPR-Cas Systems , Pluripotent Stem Cells , Humans , CRISPR-Cas Systems/genetics , Cell Line , Homologous Recombination , Cell Differentiation/physiology , TubulinABSTRACT
Protein assemblies have drawn much attention as platforms for biomedical applications, including gene/drug delivery and vaccine, due to biocompatibility and functional diversity. Here, the construction and functionalization of a protein assembly composed of human clathrin heavy chain and light chain for a targeted protein delivery, is presented. The clathrin heavy and light chains are redesigned and associated with each other, and the resulting triskelion unit further self-assembled into a clathrin assembly with the size of about 28 nm in diameter. The clathrin assembly is dual-functionalized with a protein cargo and a targeting moiety using two different orthogonal protein-ligand pairs through one-pot reaction. The functionalized clathrin assembly exhibits about a 900-fold decreased KD value for a cell-surface target due to avidity compared to a native targeting moiety. The utility of the clathrin assembly is demonstrated by an efficient delivery of a protein cargo into tumor cells in a target-specific manner, resulting in a strong cytotoxic effect. The present approach can be used in the creation of protein assemblies with multimodality.
Subject(s)
Clathrin , Drug Delivery Systems , Humans , Clathrin/metabolismABSTRACT
Mutations in myelodysplasia-related (MR) genes, rather than morphological features, have been included in the diagnostic criteria of the new 5th World Health Organization (WHO) classification for myelodysplastic syndrome (MDS)-associated acute myeloid leukemia (AML). This study compares the clinical relevance of the new criteria with those of the previous version. In a cohort of 135 patients with newly diagnosed AML, the MDS-related AML patients were classified according to the 5th and 4th edition of the WHO classification (AML, myelodysplasia-related [AML-MR5th] and AML with myelodysplasia-related changes [AML-MRC4th], respectively). The median age of the patients was 70.4 years. MR gene mutations were found in 48 patients (35.6%). Sixty-one patients (46.6%) were diagnosed with AML-MRC4th, while 71 patients (53.0%) were diagnosed with AML-MR5th. Patients with AML-MR5th were significantly older with significantly lower treatment response rate, higher recurrence rate, and shorter relapse-free survival after chemotherapy, whereas AML-MRC4th patients did not show any association with the treatment outcome. Overall, the following prognostic factors for survival were identified: age over 75 years, antecedent MDS or MDS/myeloproliferative neoplasm, chromosome 5 or 7 abnormalities, and KRAS and ZSZR2 mutations. The 5th WHO classification is more useful for predicting the treatment response of patients with AML-MR than the previous version. Among the MR genes, ZSZR2 mutations were found to be independent prognostic factors affecting survival.
Subject(s)
Leukemia, Myeloid, Acute , Myelodysplastic Syndromes , Humans , Aged , Myelodysplastic Syndromes/diagnosis , Myelodysplastic Syndromes/genetics , Myelodysplastic Syndromes/drug therapy , Leukemia, Myeloid, Acute/diagnosis , Leukemia, Myeloid, Acute/genetics , Leukemia, Myeloid, Acute/drug therapy , Mutation , Cohort Studies , World Health OrganizationABSTRACT
eXplainable Artificial Intelligence (XAI) is a new trend of machine learning. Machine learning models are used to predict or decide something, and they derive output based on a large volume of data set. Here, the problem is that it is hard to know why such prediction was derived, especially when using deep learning models. It makes the models unreliable in the case of reliability-critical applications. So, it is required to explain how they derived such output. It is a reliability-critical application for self-driving cars because the mistakes made by the computers inside them can lead to critical accidents. So, it is necessary to adopt XAI models in this field. In this paper, we propose an XAI method based on computing and explaining the difference of the output values of the neurons in the last hidden layer of convolutional neural networks. First, we input the original image and some modified images of it. Then we derive output values for each image and compare these values. Then, we introduce the Sensitivity Analysis technique to explain which parts of the original image are needed to distinguish the category. In detail, we divide the image into several parts and fill these parts with shades. First, we compute the influence value on the vector indicating the last hidden layer of the model for each of these parts. Then we draw shades whose darkness is in proportion to the influence values. The experimental results show that our approach for XAI in self-driving cars finds the parts needed to distinguish the category of these images accurately.
Subject(s)
Artificial Intelligence , Autonomous Vehicles , Algorithms , Neural Networks, Computer , Reproducibility of ResultsABSTRACT
The assembly of proteins in a programmable manner provides insight into the creation of novel functional nanomaterials for practical applications. Despite many advances, however, a rational protein assembly with an easy scalability in terms of size and valency remains a challenge. Here, a simple bottom-up approach to the supramolecular protein assembly with a tunable size and valency in a programmable manner is presented. The dendrimer-like protein assembly, simply called a "protein dendrimer," is constructed through a stepwise and alternate addition of a building block protein. Starting from zeroth-generation protein dendrimer (pG0 ) of 27 kDa, the protein dendrimer is sequentially grown to pG1 , pG2 , pG3 , to pG4 with a molecular mass of 94, 216, 483, and 959 kDa, respectively. The valency of the protein dendrimers at the periphery increases by a factor of two after each generation, allowing a tunable valency and easy functionalization. The protein dendrimers functionalizes with a targeting moiety and a cytotoxic protein cargo shows a typical feature of multi-valency in the avidity and a highly enhanced cellular cytotoxicity, exemplifying their utility as a protein delivery platform. The present approach can be effectively used in the creation of protein architectures with new functions for biotechnological and medical applications.
Subject(s)
Antineoplastic Agents/administration & dosage , Dendrimers/metabolism , Neoplasms/drug therapy , Cell Line, Tumor , Cells, Cultured , Humans , Microscopy, Confocal , Nanostructures , Neoplasms/diagnostic imagingABSTRACT
Improving drought stress tolerance of soybean could be an effective way to minimize the yield reduction in the drought prevailing regions. Identification of drought tolerance-related quantitative trait loci (QTLs) is useful to facilitate the development of stress-tolerant varieties. This study aimed to identify the QTLs for drought tolerance in soybean using a recombinant inbred line (RIL) population developed from the cross between a drought-tolerant 'PI416937' and a susceptible 'Cheonsang' cultivar. Phenotyping was done with a weighted drought coefficient derived from the vegetative and reproductive traits. The genetic map was constructed using 2648 polymorphic SNP markers that distributed on 20 chromosomes with a mean genetic distance of 1.36 cM between markers. A total of 10 QTLs with 3.52-4.7 logarithm of odds value accounting for up to 12.9% phenotypic variance were identified on seven chromosomes. Five chromosomes-2, 7, 10, 14, and 20-contained one QTL each, and chromosomes 1 and 19 harbored two and three QTLs, respectively. The chromosomal locations of seven QTLs overlapped or located close to the related QTLs and/or potential candidate genes reported earlier. The QTLs and closely linked markers could be utilized in maker-assisted selection to accelerate the breeding for drought tolerance in soybean.
ABSTRACT
Previously, we reported that the administration of a mixture of Humulus japonicus (MH) increased the longitudinal bone growth rate in Sprague Dawley rats. In this study, we investigated the effects of the dietary administration of MH on longitudinal bone growth in growth hormone (GH)-deficient hypophysectomized male and female rats to determine whether the effect of MH was similar to that of GH. We measured the nose-to-anus and nose-to-tail length gain, femur and tibia lengths, growth plate zones, and expression of insulin-like growth factor-1 (IGF-1) and IGF-binding protein-3 (IGFBP-3) after the dietary administration of MH or the injection of GH into hypophysectomized rats for 4 weeks. Results demonstrated that the dietary administration of MH had no effect on longitudinal bone growth, whereas the injection of GH increased the nose-to-tail length gain and femur and tibia lengths in hypophysectomized rats. In addition, MH did not affect the growth plate, bone mineralization, and expression of IGF-1 and IGFBP-3. These findings indicate that MH does not exert a GH-like effect and that the effects of MH and GH on longitudinal bone growth involve different pathways.
Subject(s)
Humulus , Animals , Bone Development , Female , Growth Hormone , Hypophysectomy , Insulin-Like Growth Factor I/genetics , Male , Rats , Rats, Sprague-DawleyABSTRACT
To prevent global warming, ESS development is in progress along with the development of electric vehicles and renewable energy. However, the state-of-the-art technology, i.e., lithium-ion batteries, has reached its limitation, and thus the need for high-performance batteries with improved energy and power density is increasing. Lithium-sulfur batteries (LSBs) are attracting enormous attention because of their high theoretical energy density. However, there are technical barriers to its commercialization such as the formation of dendrites on the anode and the shuttle effect of the cathode. To resolve these issues, a boron nitride nanotube (BNNT)-based separator is developed. The BNNT is physically purified so that the purified BNNT (p-BNNT) has a homogeneous pore structure because of random stacking and partial charge on the surface due to the difference of electronegativity between B and N. Compared to the conventional polypropylene (PP) separator, the p-BNNT loaded PP separator prevents the dendrite formation on the Li metal anode, facilitates the ion transfer through the separator, and alleviates the shuttle effect at the cathode. With these effects, the p-BNNT loaded PP separators enable the LSB cells to achieve a specific capacity of 1429 mAh/g, and long-term stability over 200 cycles.
ABSTRACT
The aim of this study was to investigate the effects of administration of a mixture of Humulus japonicus (MH) on longitudinal bone growth in normal Sprague Dawley (SD) rats. We measured the femur and tibia length, growth plate area, proliferation of chondrocytes, and expression of insulin-like growth factor-1 (IGF-I) and IGF binding protein-3 (IGFBP-3), and Janus kinase 2 (JAK2)/signal transducer and activator of transcription 5 (STAT5) phosphorylation after dietary administration of MH in SD rats for four weeks. The nose-tail length gain and length of femur and tibia increased significantly in the group that received MH for a period of four weeks. We performed H&E staining and Bromodeoxyuridine/5-Bromo-2'-Deoxyuridine (BrdU) staining to examine the effect of dietary administration of MH on the growth plate and the proliferation of chondrocytes and found that MH stimulated the proliferation of chondrocytes and contributed to increased growth plate height during the process of longitudinal bone growth. In addition, serum levels of IGF-1 and IGFBP-3 and expression of IGF-1 and IGFBP-3 mRNAs in the liver and bone were increased, and phosphorylation of JAK2/STAT5 in the liver was increased in the MH groups. Based on these results, we suggest that the effect of MH on longitudinal bone growth is mediated by increased JAK2/STAT5-induced IGF-1 production.
Subject(s)
Bone Development/drug effects , Humulus , Plant Extracts/pharmacology , Animals , Female , Models, Animal , Rats , Rats, Sprague-DawleyABSTRACT
Pod shattering is an important reproductive process in many wild species. However, pod shattering at the maturing stage can result in severe yield loss. The objectives of this study were to discover quantitative trait loci (QTLs) for pod shattering using two recombinant inbred line (RIL) populations derived from an elite cultivar having pod shattering tolerance, namely "Daewonkong", and to predict novel candidate QTL/genes involved in pod shattering based on their allele patterns. We found several QTLs with more than 10% phenotypic variance explained (PVE) on seven different chromosomes and found a novel candidate QTL on chromosome 16 (qPS-DS16-1) from the allele patterns in the QTL region. Out of the 41 annotated genes in the QTL region, six were found to contain SNP (single-nucleotide polymorphism)/indel variations in the coding sequence of the parents compared to the soybean reference genome. Among the six potential candidate genes, Glyma.16g076600, one of the genes with known function, showed a highly differential expression levels between the tolerant and susceptible parents in the growth stages R3 to R6. Further, Glyma.16g076600 is a homolog of AT4G19230 in Arabidopsis, whose function is related to abscisic acid catabolism. The results provide useful information to understand the genetic mechanism of pod shattering and could be used for improving the efficiency of marker-assisted selection for developing varieties of soybeans tolerant to pod shattering.
ABSTRACT
Soybeans, Glycine max (L.) Merr., are among the most important food crops worldwide. Isoflavones are major bioactive phytochemicals in soybeans, and have a variety of health benefits, including antioxidative, antiatherosclerotic, antiinflammatory, and weak estrogen-like effects. The isoflavone content and composition of soybeans vary according to the cultivar and the extraction solvent conditions. Therefore, we investigated the effects of three different solvent pHs (1.0, 5.5, and 10.0) on the isoflavone, total phenolic, and total flavonoid contents and antioxidant capacities of eight soybean cultivars developed in Korea. Twelve isoflavones in soybeans were efficiently separated and identified on a reversed-phase high-performance liquid chromatography (HPLC) system. The percentage distribution of isoflavones measured by HPLC in the eight soybean cultivars at various extraction pHs decreased as follows: malonyl isoflavones (67.2% to 81.3%) > isoflavone glucosides (16.2% to 29.0%; as nonacylated form) > acetyl isoflavones (1.6% to 5.9%). The highest contents of isoflavone glucosides, malonyl derivatives, and acetyl derivatives were extracted at solvent pHs of 10.0, 1.0, and 5.5, respectively. The solvent extraction at pH 1.0 yielded a lower total isoflavone content than those at pHs 5.5 and 10.0. However, the highest total phenolic and flavonoid contents were extracted from soybeans at pH 1.0. Soybeans extracted at pH 10.0 displayed the highest antioxidant capacities in the 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) radical assay. Taken together, these results suggest that proper solvent pH adjustment is needed to maximize the extraction of targeted forms of isoflavones from soybeans. PRACTICAL APPLICATION: Soybeans contain a variety of bioactive compounds, including isoflavones, which function as antioxidants and weak phytoestrogens. Chemical and instrumental analyses can facilitate the selection of soybean cultivars with high amounts of isoflavones for soybean breeding and isoflavone-enriched product development. Proper solvent pH adjustment allows for the efficient extraction of high amounts of targeted isoflavone subgroups (acetyl and malonyl forms) from soybeans for functional food products.
Subject(s)
Chemical Fractionation/methods , Glycine max/chemistry , Isoflavones/analysis , Isoflavones/isolation & purification , Plant Extracts/analysis , Plant Extracts/isolation & purification , Antioxidants/analysis , Antioxidants/isolation & purification , Chromatography, High Pressure Liquid , Chromatography, Reverse-Phase , Flavonoids/analysis , Flavonoids/isolation & purification , Phenols/analysis , Phenols/isolation & purification , Republic of Korea , Seeds/chemistryABSTRACT
This article contains data on subsidies provided to the fisheries sector by maritime countries. The dataset is the culmination of extensive data collection efforts using peer-reviewed and grey literature, national budgets, online databases, websites and other relevant sources (e.g. OECD, World Bank and WTO), in order to estimate the scope and magnitude of global fisheries subsidies. For subsidies where we found evidence of expenditure by a country, we record the total amount alongside the source references and refer to these as 'reported' data. Where evidence is found that a country provides a subsidy but no amount reported, we estimate using various approaches and refer to these as 'modeled' data. Where evidence exists that no subsidy is provided by a country we refer to these null values as 'not found evidence of subsidy'. All amounts were converted to constant 2018 USD using 2017 exchange rates and annual Consumer Price Index averages. The final dataset of 'reported', 'modeled' and 'not found' subsidies for 2018 consists of 13 subsidy types across 152 maritime countries. The dataset, first developed in the early 2000s, now forms part of the global fisheries management infrastructure and is a central tool used by WTO negotiators. The data we provide may be used to support local, regional and global fisheries management decision-making and may have further uses when analysed in combination with other fisheries related data. Interpretation of these data can be found in the associated research article titled "Updated estimates and analysis of global fisheries subsidies" [1].
ABSTRACT
A nociceptor is an essential element in the human body, alerting us to potential damage from extremes in temperature, pressure, etc. Realizing nociceptive behavior in an electronics device remains a central issue for researchers, designing neuromorphic devices. This study proposes and demonstrates an all-oxide-based highly transparent ultraviolet-triggered artificial nociceptor, which responds in a very similar way to the human eye. The device shows a high transmittance (>65%) and very low absorbance in the visible region. The current-voltage characteristics show loop opening, which is attributed to the charge trapping/detrapping. Further, the ultraviolet-stimuli-induced versatile criteria of a nociceptor such as a threshold, relaxation, allodynia, and hyperalgesia are demonstrated under self-biased condition, providing an energy-efficient approach for the neuromorphic device operation. The reported optically controlled features open a new avenue for the development of transparent optoelectronic nociceptors, artificial eyes, and memory storage applications.
ABSTRACT
In this work, a strain modulated highly transparent and flexible ZnO/Ag-nanowires/polyethylene terephthalate optoelectronic device is developed. By utilizing the applied external strain-induced piezophototronic effects of a ZnO thin film, a UV-generated photocurrent is tuned in a wide range starting from 0.01 to 85.07 µA and it is presented in a comprehensive map. Particularly, the performance of the device is effectively enhanced 7733 times by compressive strain, as compared to its dark current in a strain-free state. The observed results are explained quantitatively based on the modulation of oxygen desorption/absorption on the ZnO surface under the influence of applied strains. The presented simple optoelectronic device can be easily integrated into existing planar structures, with potential applications in highly transparent smart windows, wearable electronics, smartphones, security communication, and so on.
ABSTRACT
Due to their outstanding physical properties, perovskite materials are considered to be promising semiconductors for next-generation optoelectronics. However, these materials are often unstable under an ambient atmosphere and ultra-violet illumination. Therefore, the construction of an air-stable visible light transparent perovskite-based ultra-violet photodetector is still highly challenging. In this study, we go beyond the conventional operation of photodetectors by utilizing the undesired hysteresis loop in the typical current-voltage characteristics of perovskites and design a (C4H9NH3)2PbBr4-based high-performance visible transparent programmable ultra-violet photodetector. The photodetector shows multiple operating levels and can switch from one level to another with a short electric pulse. The photodetector exhibits a fast response time of â¼2 ms, good responsivity of â¼32 mA W-1 and detectivity of 8.5 × 108 Jones with a low working voltage of 0.5 V. Moreover, the photodetector shows long-term stability, and the optoelectronic performance is retained under ambient conditions. This breakthrough in the controlled tunable features opens a new avenue for the development of multipurpose transparent optoelectronic devices.
ABSTRACT
We report a self-biased and transparent Cu4O3/TiO2 heterojunction for ultraviolet photodetection. The dynamic photoresponse improved 8.5 × 104% by adding silver nanowires (AgNWs) Schottky contact and maintaining 39% transparency. The current density-voltage characteristics revealed a strong interfacial electric field, responsible for zero-bias operation. In addition, the dynamic photoresponse measurement endorsed the effective holes collection by embedded-AgNWs network, leading to fast rise and fall time of 0.439 and 0.423 ms, respectively. Similarly, a drastic improvement in responsivity and detectivity of 187.5 mAW-1 and of 5.13 × 109 Jones, is observed, respectively. The AgNWs employed as contact electrode can ensure high-performance for transparent and flexible optoelectronic applications.