• Title/Summary/Keyword: protein structure

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Studies on the Myofibrillar Proteins Part 2. New Procedure for Extraction of Regulatory Proteins from Myofibrils (근원섬유단백질에 관한 연구 (제2보) 근수축 조절단백질의 새로운 정제방법)

  • Yang, Ryung;Kim, Chul-Jai;Yu, Ju-Hyun;Lee, Hyuk-Sin;Cho, Young-Dong
    • Korean Journal of Food Science and Technology
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    • v.6 no.4
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    • pp.199-208
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    • 1974
  • An attempt was made to study on new method for the extraction of the regulatory proteins from myofibrils, and the procedures for the preparation of desensitized actomyosin and for complete extraction of troponin-tropomyosin complex were developed. When myofibrils were treated through the procedures developed in this study, actomyosin obtained had no Ca-sensitivity, indicating that Ca-sensitizing protein factor had been removed completely from myofibril. Consequently, it was concluded that the procedures developed in this study were convenient to test whether Ca-sensitizing proteins has been removed or not. When Mg-activated ATPase activity of myofibril were measured, the myofibrillar ATPase turned into the actomyosin type ATPase with the progress of the treatment. This result was interpreted to show that the regulatory proteins of the myofibril seems to play a cementing role on the structure of myofibril. When supernatant containing the regulatory proteins were fractionated with $(NH_4)_2SO_4$ saturation solution, regulatory proteins, ${\alpha}-actinin$ and troponia-tropomyosin complex, could be obtained and they showed their typical phyoislogical activity which modify the actin-myosin interaction. The amount of troponin-tropomyosin complex in myofibril was 72 mg per g myofibril. This result was in good agreement with the results reported by many investigators, and therefore it was concluded that our procedures for the extraction of troponin-tropomyosin complex were desirable to study on the quantitative analysis of troponin-tropomyosin complex.

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n-Gram/2L: A Space and Time Efficient Two-Level n-Gram Inverted Index Structure (n-gram/2L: 공간 및 시간 효율적인 2단계 n-gram 역색인 구조)

  • Kim Min-Soo;Whang Kyu-Young;Lee Jae-Gil;Lee Min-Jae
    • Journal of KIISE:Databases
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    • v.33 no.1
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    • pp.12-31
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    • 2006
  • The n-gram inverted index has two major advantages: language-neutral and error-tolerant. Due to these advantages, it has been widely used in information retrieval or in similar sequence matching for DNA and Protein databases. Nevertheless, the n-gram inverted index also has drawbacks: the size tends to be very large, and the performance of queries tends to be bad. In this paper, we propose the two-level n-gram inverted index (simply, the n-gram/2L index) that significantly reduces the size and improves the query performance while preserving the advantages of the n-gram inverted index. The proposed index eliminates the redundancy of the position information that exists in the n-gram inverted index. The proposed index is constructed in two steps: 1) extracting subsequences of length m from documents and 2) extracting n-grams from those subsequences. We formally prove that this two-step construction is identical to the relational normalization process that removes the redundancy caused by a non-trivial multivalued dependency. The n-gram/2L index has excellent properties: 1) it significantly reduces the size and improves the Performance compared with the n-gram inverted index with these improvements becoming more marked as the database size gets larger; 2) the query processing time increases only very slightly as the query length gets longer. Experimental results using databases of 1 GBytes show that the size of the n-gram/2L index is reduced by up to 1.9${\~}$2.7 times and, at the same time, the query performance is improved by up to 13.1 times compared with those of the n-gram inverted index.

The Effect of Jeju Wild Ginseng Extracts on Skin Barrier via Serine-Palmitoyltransferase (제주산양산삼이 세린-팔미토일 전이효소(Serine-Palmitoyltransferase)를 통해 피부 장벽에 미치는 효과에 대한 연구)

  • Kim, Hyo Min;Lee, Jung No;Kim, Jae Moon;Kim, Sung Kyu;Park, Sung-Min
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.42 no.2
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    • pp.119-126
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    • 2016
  • Skin is the largest organ that protects the body from the external environmental factors such as smog, cigarette smoke, UV. Protective skin barrier is composed with keratinizational keratinocytes and intercellular lipids such as ceramides, cholesterols and fatty acids combined by the lamellar liquid crystal structure. In this research, we confirmed that the Jeju wild ginseng (JWG) extracts dose-dependently increased the expression of serine-palmitoyltransferase (SPT) protein which is associated with ceramide biosynthesis. In addition, emulsion containing 5% JWG extract was applied on skin of human volunteers for 2 weeks and then significantly reduced transepidermal water loss (TEWL) compared to that of control group. As a results, JWG extract increased the biosynthesis of ceramides that is the key components of the skin lipid through enhancing expression of SPT. In addition, JWG extract reduced TEWL resulting in improvement of skin barrier function. In this context, we suggest that JWG extract could be used as a skin barrier enhancer and moisturing agents in cosmetic fileds.

Anti-inflammatory Effects of Achyranthes japonica Nakai and Aralia continentalis Kitagawa Complex Fermented Extracts on LPS-stimulated RAW264.7 Macrophage (LPS로 자극된 RAW264.7 대식세포에서 우슬 및 땅두릅 복합 발효추출물의 항염증 효과)

  • Woo, Young Min;Jo, Eun Sol;Kim, Ok Ju;Lee, Young-Ho;Ahn, Mee Young;Lee, Dong-Geun;Lee, Sang-Hyeon;Ha, Jong-Myung;Kim, Andre
    • Applied Chemistry for Engineering
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    • v.30 no.4
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    • pp.479-486
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    • 2019
  • This study investigated the anti-inflammatory effects of mixed extracts of Achyranthes japonica Nakai (Aj) and Aralia continentalis Kitagawa (Ac) (ratios of 1 : 2, 1 : 3, 1 : 5, 2 : 1, 3 : 1 and 5 : 1) on RAW264.7 macrophages. Cell toxicity was determined using a cell counting kit (CCK) assay. We evaluated anti-inflammatory effects of the mixed extracts of Aj and Ac by measuring interleukin $(IL)-1{\beta}$, IL-6, and tumor necrosis factor $(TNF){\alpha}$ using an enzyme-linked immunosorbent assay (ELISA) kit assay. The mixed extracts of Aj and Ac inhibited lipopolysaccharide (LPS)-induced $IL-1{\beta}$ and $TNF{\alpha}$ in LPS-stimulated macrophages. Comparing different ratios of the mixed extracts, the 2 : 1 ratio of Aj and Ac has much more potency and inhibited the production of $TNF{\alpha}$ in LPS-induced RAW264.7 cells. The results of the present study showed that the mixed extracts of Aj and Ac have potential anti-inflammatory effects on RAW264.7 macrophages. Therefore, these extracts may be used as a good source of functional foods for the protection against inflammatory diseases.

Genetic Diversity and Relationship in Soybean MDP (Mutant Diversity Pool) Revealed by TRAP and TE-TRAP Markers

  • Kim, Dong-Gun;Bae, Chang-Hyu;Kwon, Soon-Jae
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.04a
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    • pp.32-32
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    • 2019
  • Mutation breeding is the useful tool to improve agronomic traits in various crop species. Soybean is most important crop and is rich in protein and oil contents. Despite of the importance as economic value and various genetic resource of soybean, there have been limited studies of genetic relationship among mutant resources through radiation breeding. In this study, the agronomical phenotype for selecting various genetic resources was evaluated in 528 soybean mutant lines. As a result, 210 soybean mutants with their original cultivars were selected with various traits. We named 210 selected lines as Mutant Diversity Pool (MDP). The genetic diversity and the relationship of the MDP were investigated using TRAP and TE-TRAP markers. In TRAP analysis, sixteen primer combination (PC)s were used and a total of 551 fragments were amplified. The highest (84.00%) and the lowest (32.35%) polymorphism levels were showed in PC MIR157B+Ga5 and B14G14B+Ga3, respectively. The mean of PIC values was 0.15 ranging from 0.07 in B14G14B+Sa12 to 0.23 in MIR157B+Sa4. Phylogenetic and population structure analysis indicated that the 210 MDP lines dispersed to four groups among the wild types and their mutants. The highest genetic diversity among populations was observed between lines Paldal and 523-7 (Fst=0.409), whereas the lowest genetic diversity was between population KAS360-22 and 94seori (Fst=0.065). AMOVA showed 11.583 (21.0%) and 43.532 (79.0%) variations in inter and intra mutant population, respectively. Overall, the genetic similarity of each intra mutant populations was closer than that of inter mutant population. A total of 408 fragments were amplified in the 210 MDP using twelve PCs of TE-TRAP markers that were obtained from a combination of three TIR sequence of transposable elements (MITE-stowaway; M-s, MITE-tourist; M-t, PONG). The highest (77.42%) and the lowest (56.00%) polymorphism levels were showed in PONG+Sa4 and PONG+Sa12, respectively. The mean of PIC values was 0.15 ranging from 0.09 in M-s+Sa4 and M-s+Ga5 to 0.21 in M-t+Ga5. AMOVA of M-s showed 2.209 (20%) and 8.957 (80%) variations in inter and intra mutant population, respectively. AMOVA of M-t showed 2.766 (18%) and 12.385 (82%) variations in inter and intra mutant population, respectively. AMOVA of PONG showed 3.151 (29%) and 7.646 (71%) variations in inter and intra mutant population, respectively. According to our study, the PONG had higher inter mutant population and lower intra mutant population. This mean was that for aspect of radiation sensitivity, M-s and M-t showed higher mobility than that of PONG. Our results suggest that the TRAP and the TE-TRAP markers may be useful for assessing the genetic diversity and relationship among soybean MDP and help to improve our knowledge of soybean mutation/radiation breeding.

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Study on the Structural Analysis and Characterization of Hunting Pit in the Bronze Age - Focusing on Relics in Ulsan and Chuncheon Area - (청동기시대 함정유구의 구조분석과 성격 검토 - 울산, 춘천 지역 유적을 중심으로 -)

  • Choi, Su-hyoung
    • Korean Journal of Heritage: History & Science
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    • v.46 no.2
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    • pp.166-185
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    • 2013
  • After analyzing the location, arrangement, inner structure, and form of the hunting pit in the Bronze Age, this paper sets up three types and talks about types of hunting pits. After analyzing the types of the hunting pit, three types of hunting pits are derived: mountain district - large group's arrangement - long oval - I II type(A), mountain district - arrangement in a line - oval - I type(B), flat area - arrangement in a line - oval - I III type(C). Literature, the ways in hunting pits, the purpose of the installations, and the characters are studied and compared with those of the anthropological cases. Even though hunting pits of types A and B are constructed to hide and get protein and bone of animals for breeding, there are differences in the ways in hunting and characters. Type of A seems to be constructed only for occupation that is the standardized hunting method on a small works. However, type B is the chasing method to hunt as a group and it has various purposes including main occupation. For example, it is for improving the war skills by practicing the hunting strategies and cooperating with the people. In addition, it is for getting a sacrifice for god. The type C hunting pit is estimated that it plays a role to protect themselves from invasion. It's inferred that there are many purposes for military defense and protection of people or animals within the village. It is the reason why the construction is spread in hunting pit including various purposes in the Bronze age that originates in social and economical specialization like the increase in agricultural productivity and appearance of a chief.

Structural Disorganization of Intestinal Tumor Spheroid by Microbial Ribotoxins (방사선 모사 미생물 유래 리보솜 스트레스에 의한 대장암 스페로이드 구조 결함 유발)

  • Kim, Juil;Kim, Joongkon;Yu, Mira;Moon, Yuseok
    • Microbiology and Biotechnology Letters
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    • v.47 no.1
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    • pp.164-171
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    • 2019
  • Radiation therapy has many side effects, such as digestive mucosal ulcers, without regard to its efficacy. The purpose of this study is to address an alternative method to replace the limitation of radiation therapy using radiomimetic microbial ribotoxins. In the evaluation of cancer therapy, we analyzed the formation of colorectal cancer (CRC) cell spheroids, which can take into account the heterogeneous cellular constitution, tumor stem cells, and the surrounding microenvironment. Ribotoxic stress interfered with the spheroid structure composed of relatively small clusters. Spheroids under ribotoxic stress were structurally sparse and their shrinkage was very slow. In the control group, the clusters of strongly aggregated cells were resistant to physical stress, but the ribotoxic stress-exposed spheroids were easily broken up by the physical stress. Moreover, the ribosome-insulted CRC cells slowly migrated to form clusters and the cell-cell junctional points in the ribosome-insulted spheroids were rarer than those in the control CRC spheroid. Moreover, levels of the cell-to-cell junctional protein E-cadherin were suppressed by ribotoxic stress in both allograft and xenograft spheroids. In conclusion, the radiomimetic microbial ribotoxins induced structural defects in CRC cell spheroids via retardation of migration and cell-cell junction in the formation of three-dimensional structures, and provides a basis for the mechanism of pharmacological radiomimetic anticancer actions as an alternate to radiotherapy against cancer.

Coronaviruses: SARS, MERS and COVID-19 (코로나바이러스: 사스, 메르스 그리고 코비드-19)

  • Kim, Eun-Joong;Lee, Dongsup
    • Korean Journal of Clinical Laboratory Science
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    • v.52 no.4
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    • pp.297-309
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    • 2020
  • Coronaviruses were originally discovered as enzootic infections that limited to their natural animal hosts, but some strains have since crossed the animal-human species barrier and progressed to establish zoonotic diseases. Accordingly, cross-species barrier jumps resulted in the appearance of SARS-CoV, MERS-CoV, and SARS-CoV-2 that manifest as virulent human viruses. Coronaviruses contain four main structural proteins: spike, membrane, envelope, and nucleocapsid protein. The replication cycle is as follows: cell entry, genome translation, replication, assembly, and release. They were not considered highly pathogenic to humans until the outbreaks of SARS-CoV in 2002 in Guangdong province, China. The consequent outbreak of SARS in 2002 led to an epidemic with 8,422 cases, and a reported worldwide mortality rate of 11%. MERS-CoVs is highly related to camel CoVs. In 2019, a cluster of patients infected with 2019-nCoV was identified in an outbreak in Wuhan, China, and soon spread worldwide. 2019-nCoV is transmitted through the respiratory tract and then induced pneumonia. Molecular diagnosis based on upper respiratory region swabs is used for confirmation of this virus. This review examines the structure and genomic makeup of the viruses as well as the life cycle, diagnosis, and potential therapy.

Whitening and Anti-oxidative Constituents from the Extracts of Hydrangea petiolaris Leaves (등수국 잎 추출물 유래 미백 및 항산화 활성 성분)

  • Jo, Seong Mi;Kim, Jung Eun;Lee, Nam Ho
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.48 no.2
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    • pp.123-134
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    • 2022
  • In this study, the whitening and antioxidant effects of the extracts from Hydrangea petiolaris (H. petiolaris) leaves was confirmed, and the chemical structure was identified by separating the active ingredients. In the whitening tests using α-MSH stimulated B16F10 melanoma cells, the n-hexane (Hex) fraction inhibited the cellular melanogenesis and intracellular tyrosinase activities without causing cell toxicity. In addition, the Hex fraction reduced expression of tyrosinase and TRP-2 protein. Upon the anti-oxidative studies by DPPH and ABTS+ radicals, potent radical scavenging activities were observed in the ethyl acetate (EtOAc) fraction. Also, for the cellular protective effects on HaCat keratinocytes damaged by H2O2, the EtOAc fraction indicated protective effects against oxidative stress. Eight phytochemicals were isolated from the extract of H. petiolaris leaves; ethyl linoleate (1), ethyl linolenate (2), 1-linoleoyl glycerol (3), 1-linolenoyl glycerol (4), epi-catechin (5), afzelin (6), quercitrin (7), hyperin (8). Among the isolates, the compounds 5 - 8 showed DPPH and ABTS+ radical scavenging activities. The contents of quercitirin, a major isolated in this extract, determined by HPLC analysis were confirmed to be about 31.3 mg/g for the 70% ethanol extract and 169.8 mg/g for the EtOAc fraction. Based on these results, it was suggested that the extract from H. petiolaris leaves could be potentially applicable as whitening and anti-oxidative ingredients in cosmetic formulations.

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.