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Severe choline deficiency induces alternative splicing aberrance in optimized duck primary hepatocyte cultures

  • Zhao, Lulu;Cai, Hongying;Wu, Yongbao;Tian, Changfu;Wen, Zhiguo;Yang, Peilong
    • Animal Bioscience
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    • v.35 no.11
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    • pp.1787-1799
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    • 2022
  • Objective: Choline deficiency, one main trigger for nonalcoholic fatty liver disease (NAFLD), is closely related to lipid metabolism disorder. Previous study in a choline-deficient model has largely focused on gene expression rather than gene structure, especially sparse are studies regarding to alternative splicing (AS). In modern life science research, primary hepatocytes culture technology facilitates such studies, which can accurately imitate liver activity in vitro and show unique superiority. Whereas limitations to traditional hepatocytes culture technology exist in terms of efficiency and operability. This study pursued an optimization culture method for duck primary hepatocytes to explore AS in choline-deficient model. Methods: We performed an optimization culture method for duck primary hepatocytes with multi-step digestion procedure from Pekin duck embryos. Subsequently a NAFLD model was constructed with choline-free medium. RNA-seq and further analysis by rMATS were performed to identify AS events alterations in choline-deficency duck primary hepatocytes. Results: The results showed E13 (embryonic day 13) to E15 is suitable to obtain hepatocytes, and the viability reached over 95% by trypan blue exclusion assay. Primary hepatocyte retained their biological function as well identified by Periodic Acid-Schiff staining method and Glucose-6-phosphate dehydrogenase activity assay, respectively. Meanwhile, genes of alb and afp and specific protein of albumin were detected to verify cultured hepatocytes. Immunofluorescence was used to evaluate purity of hepatocytes, presenting up to 90%. On this base, choline-deficient model was constructed and displayed significantly increase of intracellular triglyceride and cholesterol as reported previously. Intriguingly, our data suggested that AS events in choline-deficient model were implicated in pivotal biological processes as an aberrant transcriptional regulator, of which 16 genes were involved in lipid metabolism and highly enriched in glycerophospholipid metabolism. Conclusion: An effective and rapid protocol for obtaining duck primary hepatocytes was established, by which our findings manifested choline deficiency could induce the accumulation of lipid and result in aberrant AS events in hepatocytes, providing a novel insight into various AS in the metabolism role of choline.

Small CNN-RNN Engraft Model Study for Sequence Pattern Extraction in Protein Function Prediction Problems

  • Lee, Jeung Min;Lee, Hyun
    • Journal of the Korea Society of Computer and Information
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    • v.27 no.8
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    • pp.49-59
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    • 2022
  • In this paper, we designed a new enzyme function prediction model PSCREM based on a study that compared and evaluated CNN and LSTM/GRU models, which are the most widely used deep learning models in the field of predicting functions and structures using protein sequences in 2020, under the same conditions. Sequence evolution information was used to preserve detailed patterns which would miss in CNN convolution, and the relationship information between amino acids with functional significance was extracted through overlapping RNNs. It was referenced to feature map production. The RNN family of algorithms used in small CNN-RNN models are LSTM algorithms and GRU algorithms, which are usually stacked two to three times over 100 units, but in this paper, small RNNs consisting of 10 and 20 units are overlapped. The model used the PSSM profile, which is transformed from protein sequence data. The experiment proved 86.4% the performance for the problem of predicting the main classes of enzyme number, and it was confirmed that the performance was 84.4% accurate up to the sub-sub classes of enzyme number. Thus, PSCREM better identifies unique patterns related to protein function through overlapped RNN, and Overlapped RNN is proposed as a novel methodology for protein function and structure prediction extraction.

Sustained release of alginate hydrogel containing antimicrobial peptide Chol-37(F34-R) in vitro and its effect on wound healing in murine model of Pseudomonas aeruginosa infection

  • Shuaibing Shi;Hefan Dong;Xiaoyou Chen;Siqi Xu;Yue Song;Meiting Li;Zhiling Yan ;Xiaoli Wang ;Mingfu Niu ;Min Zhang;Chengshui Liao
    • Journal of Veterinary Science
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    • v.24 no.3
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    • pp.44.1-44.17
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    • 2023
  • Background: Antibiotic resistance is a significant public health concern around the globe. Antimicrobial peptides exhibit broad-spectrum and efficient antibacterial activity with an added advantage of low drug resistance. The higher water content and 3D network structure of the hydrogels are beneficial for maintaining antimicrobial peptide activity and help to prevent degradation. The antimicrobial peptide released from hydrogels also hasten the local wound healing by promoting epithelial tissue regeneration and granulation tissue formation. Objective: This study aimed at developing sodium alginate based hydrogel loaded with a novel antimicrobial peptide Chol-37(F34-R) and to investigate the characteristics in vitro and in vivo as an alternative antibacterial wound dressing to treat infectious wounds. Methods: Hydrogels were developed and optimized by varying the concentrations of crosslinkers and subjected to various characterization tests like cross-sectional morphology, swelling index, percent water contents, water retention ratio, drug release and antibacterial activity in vitro, and Pseudomonas aeruginosa infected wound mice model in vivo. Results: The results indicated that the hydrogel C proved superior in terms of cross-sectional morphology having uniformly sized interconnected pores, a good swelling index, with the capacity to retain a higher quantity of water. Furthermore, the optimized hydrogel has been found to exert a significant antimicrobial activity against bacteria and was also found to prevent bacterial infiltration into the wound site due to forming an impermeable barrier between the wound bed and external environment. The optimized hydrogel was found to significantly hasten skin regeneration in animal models when compared to other treatments in addition to strong inhibitory effect on the release of pro-inflammatory cytokines (interleukin-1β and tumor necrosis factor-α). Conclusions: Our results suggest that sodium alginate -based hydrogels loaded with Chol-37(F34-R) hold the potential to be used as an alternative to conventional antibiotics in treating infectious skin wounds.

Development of a Novel Science Curiosity Questionnaire through Modification and Verification of the Science Curiosity Questionnaire -Through the Analysis of Science Curiosity of Pre-Service Elementary Teachers- (과학호기심 설문지의 수정 및 검증을 통한 새로운 과학호기심 설문지의 개발 - 초등예비교사의 과학호기심 분석을 통하여 -)

  • Kim, Dong Uk;Shin, Min Hyeon
    • Journal of Korean Elementary Science Education
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    • v.42 no.1
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    • pp.149-160
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    • 2023
  • A Korean-version science curiosity questionnaire (Science Curiosity in Learning Environments [SCILE(15)]) was developed after factor analysis of the Korean-version SCILE(30) questionnaire. Pre-service elementary school teachers were surveyed using the Korean-version SCILE(30), and a factor analysis based on their responses was performed. The factor analysis demonstrated that the Korean-version SCILE(15) consisted of three curiosity factors: a 'science practices' factor, a 'stretching' factor, and an 'embracing' factor. Confirmatory factor analysis of the factor structure revealed correlations between all the factors, thus confirming their commonality as a science curiosity factors. The Cronbach alpha for the reliability of all items in the Korean-version SCILE(15) and of items by factor was greater than 0.700. The Korean-version SCILE(15) was therefore evaluated to be reliable as a science curiosity questionnaire. Pre-service elementary teachers who participated in the survey for the development of the SCILE(15) were aware of the 'science practices', 'stretching', and 'embracing' science curiosity factors. Analysis in a general linear model of the degree of recognition accorded by pre-service elementary teachers to the three science curiosity factors demonstrated significant differences between the curiosity factors in terms of recognition. This cohort of pre-service elementary teachers showed the highest level of recognition of the 'stretching' curiosity factor and the lowest level of recognition of the 'embracing' curiosity factor.

Receptor binding motif surrounding sites in the Spike 1 protein of infectious bronchitis virus have high susceptibility to mutation related to selective pressure

  • Seung-Min Hong;Seung-Ji Kim;Se-Hee An;Jiye Kim;Eun-Jin Ha;Howon Kim;Hyuk-Joon Kwon;Kang-Seuk Choi
    • Journal of Veterinary Science
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    • v.24 no.4
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    • pp.51.1-51.17
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    • 2023
  • Background: To date, various genotypes of infectious bronchitis virus (IBV) have co-circulated and in Korea, GI-15 and GI-19 lineages were prevailing. The spike protein, particularly S1 subunit, is responsible for receptor binding, contains hypervariable regions and is also responsible for the emerging of novel variants. Objective: This study aims to investigate the putative major amino acid substitutions for the variants in GI-19. Methods: The S1 sequence data of IBV isolated from 1986 to 2021 in Korea (n = 188) were analyzed. Sequence alignments were carried out using Multiple alignment using Fast Fourier Transform of Geneious prime. The phylogenetic tree was generated using MEGA-11 (ver. 11.0.10) and Bayesian analysis was performed by BEAST v1.10.4. Selective pressure was analyzed via online server Datamonkey. Highlights and visualization of putative critical amino acid were conducted by using PyMol software (version 2.3). Results: Most (93.5%) belonged to the GI-19 lineage in Korea, and the GI-19 lineage was further divided into seven subgroups: KM91-like (Clade A and B), K40/09-like, QX-like (I-IV). Positive selection was identified at nine and six residues in S1 for KM91-like and QX-like IBVs, respectively. In addition, several positive selection sites of S1-NTD were indicated to have mutations at common locations even when new clades were generated. They were all located on the lateral surface of the quaternary structure of the S1 subunits in close proximity to the receptor-binding motif (RBM), putative RBM motif and neutralizing antigenic sites in S1. Conclusions: Our results suggest RBM surrounding sites in the S1 subunit of IBV are highly susceptible to mutation by selective pressure during evolution.

Development of Simulation for Estimating Growth Changes of Locally Managed European Beech Forests in the Eifel Region of Germany (독일 아이펠의 지역적 관리에 따른 유럽너도밤나무 숲의 생장변화 추정을 위한 시뮬레이션 개발)

  • Jae-gyun Byun;Martina Ross-Nickoll;Richard Ottermanns
    • Journal of the Korea Society for Simulation
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    • v.33 no.1
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    • pp.1-17
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    • 2024
  • Forest management is known to beneficially influence stand structure and wood production, yet quantitative understanding as well as an illustrative depiction of the effects of different management approaches on tree growth and stand dynamics are still scarce. Long-term management of beech forests must balance public interests with ecological aspects. Efficient forest management requires the reliable prediction of tree growth change. We aimed to develop a novel hybrid simulation approach, which realistically simulates short- as well as long-term effects of different forest management regimes commonly applied, but not limited, to German low mountain ranges, including near-natural forest management based on single-tree selection harvesting. The model basically consists of three modules for (a) natural seedling regeneration, (b) mortality adjustment, and (c) tree growth simulation. In our approach, an existing validated growth model was used to calculate single year tree growth, and expanded on by including in a newly developed simulation process using calibrated modules based on practical experience in forest management and advice from the local forest. We included the following different beech forest-management scenarios that are representative for German low mountain ranges to our simulation tool: (1) plantation, (2) continuous cover forestry, and (3) reserved forest. The simulation results show a robust consistency with expert knowledge as well as a great comparability with mid-term monitoring data, indicating a strong model performance. We successfully developed a hybrid simulation that realistically reflects different management strategies and tree growth in low mountain range. This study represents a basis for a new model calibration method, which has translational potential for further studies to develop reliable tailor-made models adjusted to local situations in beech forest management.

Radiologic assessment of the optimal point for tube thoracostomy using the sternum as a landmark: a computed tomography-based analysis

  • Jaeik Jang;Jae-Hyug Woo;Mina Lee;Woo Sung Choi;Yong Su Lim;Jin Seong Cho;Jae Ho Jang;Jea Yeon Choi;Sung Youl Hyun
    • Journal of Trauma and Injury
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    • v.37 no.1
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    • pp.37-47
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    • 2024
  • Purpose: This study aimed at developing a novel tube thoracostomy technique using the sternum, a fixed anatomical structure, as an indicator to reduce the possibility of incorrect chest tube positioning and complications in patients with chest trauma. Methods: This retrospective study analyzed the data of 184 patients with chest trauma who were aged ≥18 years, visited a single regional trauma center in Korea between April and June 2022, and underwent chest computed tomography (CT) with their arms down. The conventional gold standard, 5th intercostal space (ICS) method, was compared to the lower 1/2, 1/3, and 1/4 of the sternum method by analyzing CT images. Results: When virtual tube thoracostomy routes were drawn at the mid-axillary line at the 5th ICS level, 150 patients (81.5%) on the right side and 179 patients (97.3%) on the left did not pass the diaphragm. However, at the lower 1/2 of the sternum level, 171 patients (92.9%, P<0.001) on the right and 182 patients (98.9%, P= 0.250) on the left did not pass the diaphragm. At the 5th ICS level, 129 patients (70.1%) on the right and 156 patients (84.8%) on the left were located in the safety zone and did not pass the diaphragm. Alternatively, at the lower 1/2, 1/3, and 1/4 of the sternum level, 139 (75.5%, P=0.185), 49 (26.6%, P<0.001), and 10 (5.4%, P<0.001), respectively, on the right, and 146 (79.3%, P=0.041), 69 (37.5%, P<0.001), and 16 (8.7%, P<0.001) on the left were located in the safety zone and did not pass the diaphragm. Compared to the conventional 5th ICS method, the sternum 1/2 method had a safety zone prediction sensitivity of 90.0% to 90.7%, and 97.3% to 100% sensitivity for not passing the diaphragm. Conclusions: Using the sternum length as a tube thoracostomy indicator might be feasible.

Bovine mastitis-associated Escherichia coli

  • Hong Qui Le;Se Kye Kim;Jang Won Yoon
    • Journal of Food Hygiene and Safety
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    • v.39 no.3
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    • pp.181-190
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    • 2024
  • Bovine mastitis-associated Escherichia coli (BMEC) is considered the main causative agent of significant financial losses in the dairy industry worldwide, as it alters both the quantity and quality of milk produced and increases the rate of culling. This creates a variety of challenges for researchers, veterinarians, and farmers in understanding and determining the most effective therapies and diagnostic techniques. Subclinical mastitis is particularly concerning, as infected bovines exhibit no obvious symptoms and continue to secrete apparently normal milk over an extended period, allowing the causative pathogen, E. coli, to spread within the herd. For effective prevention, understanding the pathogenesis of mastitis through three stages invasion, infection, and inflammation is essential. To date, no clear correlation has been found between virulence factors and pathogenicity contributing to the clinical severity of BMEC. Multidrug-resistant E. coli and the evolution of novel resistance mechanisms have become concerns owing to the extensive use of antibiotics to treat mastitis. Therefore, it is vital to explore alternative controls to enhance the efficacy of BMEC treatment. Over the past 30 years, various genetic typing techniques have been used to examine the subspecies-level epidemiology of bovine mastitis. These studies have advanced our understanding of the origin, transmission pathway, population structure, and evolutionary relatedness of BMEC strains. In this review we provide an overview of BMEC, including insights into its etiology, genetic relationship, pathogenesis, and management of the disease, as well as new therapy options.

Energy build-up factors estimation for BaZr0.10Ti0.90O3, Ba0.90La0.10TiO3 and Ba0.90La0.10Zr0.10Ti0.90O3 ceramics in shielding applications

  • Sarabjeet Kaur;Vidushi Karol;Pankaj Kumar;Gurpreet Kaur;Prianka Sharma;Amandeep Saroa;Amrit Singh
    • Nuclear Engineering and Technology
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    • v.56 no.5
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    • pp.1822-1829
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    • 2024
  • The search for materials that serve as good shields for radiation has become very important in light of the increasing exposure to ionizing radiation in various vital sectors. The aim is to search for novel materials with better radiation shielding properties that are stable, nontoxic, and abundant and environment friendly. The solidstate reaction approach has been used to synthesize a few ceramics, including BaZrXTi1-XO3, Ba1-XLaXTiO3 and Ba1-XLaXZrXTi1-XO3 (with x = 0.10) i.eBaZr0.10Ti0.90O3 (BZT), Ba0.90La0.10TiO3 (BLT), and Ba0.90La0.10Zr0.10Ti0.90O3 (BLZT). The density of the prepared samples varies from 6.3471 to 11.6003 g/cm3. The X-ray diffraction technique, shows strong peaks to confirm the crystalline structure of prepared ceramic samples. Using the G-P fitting approach, the advanced radiation shielding parameters (build-up factor) have been evaluated in the photon energy region of 1.5 keV-15 MeV. It is observed from the results that exposure buildup factor (EBF) and energy absorption buildup factor (EABF) are maximum for BLZT and has the minimum value for BZT in the entire photon energy regime. The results of this work should be useful in radiation shielding applications such as in industry, medicine, and nuclear engineering.

Extracting Beginning Boundaries for Efficient Management of Movie Storytelling Contents (스토리텔링 콘텐츠의 효과적인 관리를 위한 영화 스토리 발단부의 자동 경계 추출)

  • Park, Seung-Bo;You, Eun-Soon;Jung, Jason J.
    • Journal of Intelligence and Information Systems
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    • v.17 no.4
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    • pp.279-292
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    • 2011
  • Movie is a representative media that can transmit stories to audiences. Basically, a story is described by characters in the movie. Different from other simple videos, movies deploy narrative structures for explaining various conflicts or collaborations between characters. These narrative structures consist of 3 main acts, which are beginning, middle, and ending. The beginning act includes 1) introduction to main characters and backgrounds, and 2) conflicts implication and clues for incidents. The middle act describes the events developed by both inside and outside factors and the story dramatic tension heighten. Finally, in the end act, the events are developed are resolved, and the topic of story and message of writer are transmitted. When story information is extracted from movie, it is needed to consider that it has different weights by narrative structure. Namely, when some information is extracted, it has a different influence to story deployment depending on where it locates at the beginning, middle and end acts. The beginning act is the part that exposes to audiences for story set-up various information such as setting of characters and depiction of backgrounds. And thus, it is necessary to extract much kind information from the beginning act in order to abstract a movie or retrieve character information. Thereby, this paper proposes a novel method for extracting the beginning boundaries. It is the method that detects a boundary scene between the beginning act and middle using the accumulation graph of characters. The beginning act consists of the scenes that introduce important characters, imply the conflict relationship between them, and suggest clues to resolve troubles. First, a scene that the new important characters don't appear any more should be detected in order to extract a scene completed the introduction of them. The important characters mean the major and minor characters, which can be dealt as important characters since they lead story progression. Extra should be excluded in order to extract a scene completed the introduction of important characters in the accumulation graph of characters. Extra means the characters that appear only several scenes. Second, the inflection point is detected in the accumulation graph of characters. It is the point that the increasing line changes to horizontal line. Namely, when the slope of line keeps zero during long scenes, starting point of this line with zero slope becomes the inflection point. Inflection point will be detected in the accumulation graph of characters without extra. Third, several scenes are considered as additional story progression such as conflicts implication and clues suggestion. Actually, movie story can arrive at a scene located between beginning act and middle when additional several scenes are elapsed after the introduction of important characters. We will decide the ratio of additional scenes for total scenes by experiment in order to detect this scene. The ratio of additional scenes is gained as 7.67% by experiment. It is the story inflection point to change from beginning to middle act when this ratio is added to the inflection point of graph. Our proposed method consists of these three steps. We selected 10 movies for experiment and evaluation. These movies consisted of various genres. By measuring the accuracy of boundary detection experiment, we have shown that the proposed method is more efficient.