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Isolation and Characterization of a Rice Mitochondrial Small Heat Shock Protein Gene

  • Kim, Do-Hyun;Alam, Iftekhar;Lee, Dong-Gi;Lee, Byung-Hyun
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.40 no.4
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    • pp.285-290
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    • 2020
  • To understand the role of small heat shock protein (sHSPs) in rice plant response to various stresses such as the heat and oxidative stresses, a cDNA encoding a 24.1 kDa mitochondrial small HSP (Oshsp24.1) was isolated from rice by rapid amplification of cDNA ends (RACE) PCR. The deduced amino acid sequence shows very high similarity with other plant small HSPs. DNA gel blot analysis suggests that the rice genome contains more than one copy of Oshsp24.1. High level of expression of Oshsp24.1 transcript was observed in rice seedlings in response to heat, methyl viologen, hydrogen peroxide, ozone, salt and heavy metal stresses. Recombinant OsHSP24.1 protein was produced in E. coli cells for biochemical assay. The protein formed oligomeric complex when incubated with Sulfo-EGS (ethylene glycol bis (succinimidyl succinate)). Our results shows that Oshsp24.1 has an important role in abiotic stress response and have potential for developing stress-tolerant plants.

27-Hydroxycholesterol induces macrophage gene expression via LXR-dependent and -independent mechanisms

  • Kim, Bo-Young;Son, Yonghae;Cho, Hyok-rae;Lee, Dongjun;Eo, Seong-Kug;Kim, Koanhoi
    • The Korean Journal of Physiology and Pharmacology
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    • v.25 no.2
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    • pp.111-118
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    • 2021
  • 27-Hydroxycholesterol (27OHChol) exhibits agonistic activity for liver X receptors (LXRs). To determine roles of the LXR agonistic activity in macrophage gene expression, we investigated the effects of LXR inhibition on the 27OHChol-induced genes. Treatment of human THP-1 cells with GSK 2033, a potent cell-active LXR antagonist, results in complete inhibition in the transcription of LXR target genes (such as LXRα and ABCA1) induced by 27OHChol or a synthetic LXR ligand TO 901317. Whereas expression of CCL2 and CCL4 remains unaffected by GSK 2033, TNF-α expression is further induced and 27OHChol-induced CCL3 and CXCL8 genes are suppressed at both the transcriptional and protein translation levels in the presence of GSK 2033. This LXR antagonist downregulates transcript levels and surface expression of CD163 and CD206 and suppresses the transcription of CD14, CD80, and CD86 genes without downregulating their surface levels. GSK 2033 alone had no effect on the basal expression levels of the aforementioned genes. Collectively, these results indicate that LXR inhibition leads to differential regulation of 27-hydroxycholesterol-induced genes in macrophages. We propose that 27OHChol induces gene expression and modulates macrophage functions via LXR-dependent and -independent mechanisms.

Technological Factors Facilitating B40's Motivation in Malaysia to Continue Using Online Crowdsourcing Platform

  • NA'IN, Nuramalina;HUSIN, Mohd Heikal;BAHARUDIN, Ahmad Suhaimi
    • The Journal of Asian Finance, Economics and Business
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    • v.8 no.8
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    • pp.117-126
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    • 2021
  • The increasing number of retrenchments because of the current global pandemic, Covid-19, has led many to shift to the digital economy, especially among the low-income group (B40) in Malaysia. Crowdsourcing is the collection of information, opinions, or work from a group of people, usually sourced via the Internet. Fueled by the development of Internet-based platforms that provided its technological foundation, and the need for an agile and uniquely skilled workforce, crowdsourcing has grown from the grassroots, with a burgeoning body of research investigating its many aspects. However, very few studies examined crowd workers' motivation for continuous participation on online crowdsourcing platforms. Thus, this paper aims to explore the technological factors that facilitate B40's group motivation in Malaysia to continue to participate in online crowdsourcing platforms. This paper employed a qualitative approach, using a semi-structured interview. The thematic analysis method was used to decode the data extracted from the interview transcript. The finding of this study identified four main themes and seven sub-themes: (1) Technology efficacy, (2) Platform Management: client-worker management, safety net, payment mechanism, (3) Platform Design: UI design, rating feature and (4) Infrastructure: Internet connection, technology infrastructure. This study can provide a guideline for managing crowdsourcing practices in Malaysia, especially for the crowdsourcing platform developer.

Breastfeeding experiences of women with gestational diabetes (임신성 당뇨 산모의 모유수유 이행 경험)

  • Park, Seungmi;Yu, Soo-Young
    • The Journal of Korean Academic Society of Nursing Education
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    • v.27 no.3
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    • pp.274-286
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    • 2021
  • Purpose: This study aimed to explore the lived experiences of breastfeeding women with gestational diabetes and to understand the meaning of breastfeeding for them and its encompassing context. Methods: Qualitative data were collected by interviewing 15 mothers with gestational diabetes. The transcript data from 5 focus group interviews and 2 individual interviews were analyzed using thematic analysis. Results: A core theme and 10 sub-themes emerged. The core theme that emerged was "the maternal commitment enabled for a limited time a way to compensate for guilt." Because mothers had to control their blood sugar levels before childbirth, they were worried about whether it would harm the newborn baby and thus searched for dietary information. After the babies were born and separated from their mothers, they were free from gestational diabetes. Mothers then felt guilty about their unhealthy bodies when they were pregnant. This regret provides a motivational context for breastfeeding. However, the mothers realized that breastfeeding requires commitment and dedication and that they need breastfeeding support, such as being provided practical help. Conclusion: The results showed that the breastfeeding intention of women with gestational diabetes might originate from their feelings of guilt. Therefore, an integrated breastfeeding program that combines the empathetic support of women with similar experiences and that shares practical information from experts should be implemented in a structured and systematic way.

Analysis of Aluminum Stress-induced Differentially Expressed Proteins in Alfalfa Roots Using Proteomic Approach

  • Kim, Dong-Hyun;Lee, Joon-Woo;Min, Chang-Woo;Rahman, Md. Atikur;Kim, Yong-Goo;Lee, Byung-Hyun
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.42 no.3
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    • pp.137-145
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    • 2022
  • Aluminum (Al) is one of the major factors adversely affects crop growth and productivity in acidic soils. In this study, the effect of Al on plants in soil was investigated by comparing the protein expression profiles of alfalfa roots exposed to Al stress treatment. Two-week-old alfalfa seedlings were exposed to Al stress treatment at pH 4.0. Total protein was extracted from alfalfa root tissue and analyzed by two-dimensional gel electrophoresis combined with MALDI-TOF/TOF mass spectrometry. A total of 45 proteins differentially expressed in Al stress-treated alfalfa root tissues were identified, of which 28 were up-regulated and 17 were down-regulated. Of the differentially expressed proteins, 7 representative proteins were further confirmed for transcript accumulation by RT-PCR analysis. The identified proteins were involved in several functional categories including disease/defense (24%), energy (22%), protein destination (9%), metabolism (7%), transcription (5%), secondary metabolism (4%), and ambiguous classification (29%). The identification of key candidate genes induced by Al in alfalfa roots will be useful to elucidate the molecular mechanisms of Al stress tolerance in alfalfa plants.

Pathogenicity of a Korean isolate of Pepper mild mottle virus and development of full-length cDNA clone for infectious in vitro transcripts

  • J.Y. Yoon;Park, J.K.;Y.M. Yu;K.H. Ryu
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.143.3-144
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    • 2003
  • A Korean isolate of Pepper mild mottle virus (PMMoV-Kr) was isolated from a diseased pepper crop in Chunchon, Korea. The isolate was biologically purified on Nicoticaa tabacum cv. Xanthi-nc by successive single local transfer steps, and propagated on N. tabacum cv. Samsun. PMMoV-Kr could systemically infect on N. glauca, N. benthmiana, N. occidentalis and Lycopersicon esculentum, which is typical of known isolates of PMMoV. PMMoV-Kr belongs to the pathotype P1,2 based on pepper-tobamoviral indicator experiments; Capsicn chinone harboring L3 gene revealed resistant (necrotic local lesion on inoculated leaf, HR) whereas L+, L1 and L2 pepper plants expressed susceptible reactions of mosaic systemic symptoms for the isolate. To confirm the pathology and delineate symptom determinant of the isolate, full-length cDNAs of PMMoV-Kr were amplified by RT-PCR with a primer set corresponding to the 5'- and 3'-ends of PMMoV. The RT-PCR molecules amplified from genome RNA of the isolate was cloned into the pUC18 vector. Full-length cDNA clones constructed under the control of the T7 RNA promoter could be successfully transcribed to produce in vitro transcript RNA. Infectivity of the capped transcripts and its progeny virus was verified by Western blot and RT-PCR analyses.

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CGI-58 Protein Acts as a Positive Regulator of Triacylglycerol Accumulation in Phaeodactylum tricornutum

  • Qin Shu;Yufang Pan;Hanhua Hu
    • Journal of Microbiology and Biotechnology
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    • v.33 no.2
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    • pp.242-250
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    • 2023
  • Comparative gene identification-58 (CGI-58) is an activating protein of triacylglycerol (TAG) lipase. It has a variety of catalytic activities whereby it may play different roles in diverse organisms. In this study, a homolog of CGI-58 in Phaeodactylum tricornutum (PtCGI-58) was identified. PtCGI-58 was localized in mitochondria by GFP fusion protein analysis, which is different from the reported subcellular localization of CGI-58 in animals and plants. Respectively, PtCGI-58 overexpression resulted in increased neutral lipid content and TAG accumulation by 42-46% and 21-32%. Likewise, it also increased the relative content of eicosapentaenoic acid (EPA), and in particular, the EPA content in TAGs almost doubled. Transcript levels of genes involved in de novo fatty acid synthesis and mitochondrial β-oxidation were significantly upregulated in PtCGI-58 overexpression strains compared with wild-type cells. Our findings suggest that PtCGI-58 may mediate the breakdown of lipids in mitochondria and the recycling of acyl chains derived from mitochondrial β-oxidation into TAG biosynthesis. Moreover, this study potentially illuminates new functions for CGI-58 in lipid homeostasis and provides a strategy to enrich EPA in algal TAGs.

Identification and structure of AIMP2-DX2 for therapeutic perspectives

  • Hyeon Jin Kim;Mi Suk Jeong;Se Bok Jang
    • BMB Reports
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    • v.57 no.7
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    • pp.318-323
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    • 2024
  • Regulation of cell fate and lung cell differentiation is associated with Aminoacyl-tRNA synthetases (ARS)-interacting multifunctional protein 2 (AIMP2), which acts as a non-enzymatic component required for the multi-tRNA synthetase complex. In response to DNA damage, a component of AIMP2 separates from the multi-tRNA synthetase complex, binds to p53, and prevents its degradation by MDM2, inducing apoptosis. Additionally, AIMP2 reduces proliferation in TGF-β and Wnt pathways, while enhancing apoptotic signaling induced by tumor necrosis factor-α. Given the crucial role of these pathways in tumorigenesis, AIMP2 is expected to function as a broad-spectrum tumor suppressor. The full-length AIMP2 transcript consists of four exons, with a small section of the pre-mRNA undergoing alternative splicing to produce a variant (AIMP2-DX2) lacking the second exon. AIMP2-DX2 binds to FBP, TRAF2, and p53 similarly to AIMP2, but competes with AIMP2 for binding to these target proteins, thereby impairing its tumor-suppressive activity. AIMP2-DX2 is specifically expressed in a diverse range of cancer cells, including breast cancer, liver cancer, bone cancer, and stomach cancer. There is growing interest in AIMP2-DX2 as a promising biomarker for prognosis and diagnosis, with AIMP2-DX2 inhibition attracting significant interest as a potentially effective therapeutic approach for the treatment of lung, ovarian, prostate, and nasopharyngeal cancers.

Ameliorating effects of Cuscuta chinensis Lamak extract on hind-limb ischemia, and angiogenic- or inflammatory associated factors in ovariectomized mice

  • Hye Jin Kim;Hyun Yang;Dong Ho Jung;Joo Tae Hwang;Byoung‑Seob Ko
    • Molecular Medicine Reports
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    • v.19 no.4
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    • pp.3321-3329
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    • 2019
  • Cuscuta chinensis Lamak (CCL) has traditionally been used in Korea to treat sexual disorders and skin problems. The aim of the present study was to investigate the effects of CCL extract on surgical injury-induced ischemia in the hind limbs of mice. Specifically, female C57BL/6 mice were ovariectomized, and their hind-limb vessels were ligated with surgical silk (6-0) and excised. CCL (150 or 450 mg/kg/BW) was then administered to the mice for 3 weeks, and the blood flow rate was evaluated using a laser Doppler system at -7, 0, 7, 14 and 21 days following hind-limb ischemia. The serum expression profiles of angiogenic and inflammatory mediators were measured using an antibody array, and the transcript levels were reverse transcription-quantitative polymerase chain reaction. The rate of hind limb blood flow was normalized to non-ischemic lesions and revealed to be markedly elevated at 14 and 21 days following ischemia when compared with the vehicle group. The density of capillaries in the hind limbs was also significantly increased following treatment with CCL in a dose-dependent manner. In addition, the transcriptional expression of angiogenetic factors were upregulated, whereas that of inflammatory cytokines were downregulated. Finally, vascular endothelial cell migration and tube formation were evaluated in vitro using human umbilical vein endothelial cells (HUVECs) and identified to be significantly increased following treatment with CCL. Overall, the results of the present study indicate that CCL extract exhibits therapeutic potential for the treatment of hind-limb ischemia as it promotes peripheral angiogenic and anti-inflammatory effects in mice.

"It's easy. We got Desmos right here": The role of mathematical action technology in positioning students as mathematical explorers

  • Kristen Fye;Samantha Fletcher
    • Research in Mathematical Education
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    • v.27 no.3
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    • pp.267-294
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    • 2024
  • The positive impact on student learning and continued support of mathematical action technology (MAT) in classrooms deems a need to better understand what teaching practices maximize the affordances of MATs. The purpose of this study was to better understand the technology-centered teacher moves that allow students the opportunity to be positioned as mathematical explorers and sustain mathematical authority during a MAT task. In this case study of a MAT task designed to leverage the power of sliders in Desmos to explore key characteristics of the sine function, participants were two ninth-grade students (age 14), who engaged with a task-based interview. By coding the transcript of the task-based interview, the findings identified and described the teacher's actions with the technology that resulted in meaningful mathematical activity for the two students. Along with teacher actions with the technology, evidence showed the importance of the design of the MAT task and the ability of students to troubleshoot the technology. Ultimately, we identified important considerations for teaching mathematics with technology as well as several technology-centered teaching moves, leaving room for the students to perform as mathematical explorers. Applying these research methods for future cases could help generalize these technology-centered teaching strategies that position students as mathematical explorers, thus strengthening students' mathematics identities.